The Core Electron Microscopy Facility at Dana Farber Cancer Institute located in Boston, Massachusetts has a position immediately open for a full time EM Technician to assist in daily operation of the central research facility. Requirements include: B.S. or equivalent in life sciences; one to two years experience working as an EM technician or histology technician; familiar with scanning and transmission electron microscopy and associated preparative techniques, including tissue processing, Epoxy resin embedding, ultrathin sectioning and contrast staining, immunogold staining, and dark room procedure. Computer knowledge including word processing and data base programs is required. Cryoultramicrotomy skill is desirable but not required. Office management skills including filing, ordering and billing are also helpful. M-F, 40h/W, Salary $20-25K/year with excellent benefit. DFCI is an equal opportunity Employer. If interested, please send by email resume to Yuhui Xu, MD,PhD, at the following address: Yuhui_Xu-at-dfci.harvard.edu. Or contact by telephone at (617)632-5753. Fax (617)632-5165.
I'd like to know the ways that people use for analyzing SAD patterns from samples that are polycrystalline where the patterns are not complete rings and where there are two or more phases present. I'm interested in techniques that employ an automated method for determining the d-spacings present.
Currently, I am using a program that takes the digital SAD image and gives a circularly integrated 1-dimensional pattern that can be directly compared with X-ray diffraction patterns and either the JCPDS or NIST Crystal Databases. It works very well for complete rings, but some modifications were necessary for "spotty" patterns because the integration process doesn't take account of the geometric problems with more dark pixels in the outer radii. Right now the software has a couple of options on how to integrate "spotty" patterns, but they are kludgy.
Please drop me a line and let me know how you do them.
I have stumbled across a reference to Hoffman modulation contrast (HMC). Briefly, the article states that HMC allows 3-D like images. It is possible to view specimens through plastic dishes (a limitation of Nomarski DIC ?) and to view thick samples (a limitation of phase contrast).
Can anyone tell me more about this technique please?
Many thanks, Felicity
Felicity Lawrence
Analytical Electron Microscopy Facility Queensland University of Technology GPO Box 2434, Brisbane 4000 Australia
I have 200 MB colour images from microscopy on the hard disc in the image analyser Quantimet 570 color. Quantimet save colour images in the form image file: red, green and blue.
How print this images on the color ?
Krzysztof Jan Hubner
{hubner-at-IOd.krakow.pl}
Foundry Research Institute Research Materials Department, Head Structural and Physical Research Laboratory Zakopianska 73 Call +48 12 605022 ext. 356 30-148 KRAKOW - POLAND Fax +48 12 665478, + 48 12 660870 :-)
Ann Arbor Science Publishers, Inc. P.O. Box 1425 Ann Arbor, Michigan 48106
This reference is out of print (if memory serves me correctly) but many light microscopists have the reference. It was being sold at one time on CD-Rom.
It may be available still at the following:
Microscope Publications Ltd - Products Available from Microscope Publications Ltd. The Microscope Journal. Books in The Microscope Series. Other McRI Publications. Return to McRI Home.. --http://www.mcri.org/McRI_products.html
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello,
I have stumbled across a reference to Hoffman modulation contrast (HMC). Briefly, the article states that HMC allows 3-D like images. It is possible to view specimens through plastic dishes (a limitation of Nomarski DIC ?) and to view thick samples (a limitation of phase contrast).
Can anyone tell me more about this technique please?
Many thanks, Felicity
Felicity Lawrence
Analytical Electron Microscopy Facility Queensland University of Technology GPO Box 2434, Brisbane 4000 Australia
Since it is the lack of sufficient numbers of grains and different orientations that are resposible for spotty ring patterns, our solution has been to move the sample holder while the diffraction pattern is being exposed. We move both the X-Y drives and the tilt. This takes some practice in setting the right exposure time and speed of sample movement, but more complete rings can be obtained.
Ciao for now, Ken
} I'd like to know the ways that people use for analyzing SAD patterns from } samples that are polycrystalline where the patterns are not complete rings and } where there are two or more phases present. I'm interested in techniques } that employ an automated method for determining the d-spacings present. } } Currently, I am using a program that takes the digital SAD image and gives a } circularly integrated 1-dimensional pattern that can be directly compared with } X-ray diffraction patterns and either the JCPDS or NIST Crystal Databases. It } works very well for complete rings, but some modifications were necessary for } "spotty" patterns because the integration process doesn't take account of the } geometric problems with more dark pixels in the outer radii. Right now the } software has a couple of options on how to integrate "spotty" patterns, but } they are kludgy. } } Please drop me a line and let me know how you do them. } } - -Scott Walck
Kenneth JT Livi Department of Earth and Planetary Sciences 34th and Charles Streets The Johns Hopkins University Baltimore, Maryland 21218
I too collect SAD patterns digitally and analyze them live time by searching the PDF and EDD data. I use a couple of image processing programs to reduce the data, however, I never do anything automatically since there is often so much data in a SAD pattern, such as double diffraction or satellite spots. Therefore, I chose manually which rings or partial rings or spots I want to contribute to the pattern being searched. Be careful also when interpreting intensities in the TEM. It can be misleading. For what it is worth ...
Jim Heuer General Electric Co. (510) 862-4501 heuerj-at-vncpo1.ne.ge.com
1. Scott, save that question and ask it again in the "Meet the Experts" Diffraction session at this year's MSA meeting. We are trying something new to us at MSA (credit to the Society of Vacuum Coaters who've had this type of session for some time now). We have identified areas in the physical sciences and separately in the biological sciences where people occasionally need some answers. Electron diffraction is one of the areas to be addressed in this year's meeting, chaired by Alwyn Eades. Essentially we will have an empty room with a slide and transparency projector in place. Alwyn, and anyone else Alwyn chooses to join him, sits up front, people come in, take a seat and ask questions. Either Alwyn or someone else in the room is likely to know the answer. The room is scheduled for an hour. Your question is exactly what we expect to have asked.
The other two physical science sessions confirmed are 'vacuum' with Wil Bigelow and 'phys sci specimen preparation' with Reza Alani and me. We are also considering a session on 'phys sci technologist training' but haven't firmed this up yet. Joe Mascorro is responsible overall for the biological side of this and I'm doing the physical side. We plan to put more detailed info on MSA's WWW server soon.
2. I prefer to manually pick out diffraction spots and partial rings as belonging to a particular phase in a mixture. I might take a diff pattern of a specimen that is being translated during exposure to bring more grains into the aperture to get a better grasp on which phase is which and some idea of intensities (I know, intensities are unreliable but in the absence of very strong texture, strong is strong and weak is weak and this info often helps solve phases)--but I wouldn't try to take d-spacings from a translated pattern. For accurate d-spacings the entire process has to have a precision of 1% or better. This is do-able but not easy. Electronic (video/CCD cameras,...) data collection is marginal at the 1% precision level in my experience with photographic methods ranging in precision from "adequate" to "poor." Instrumental techniques, like making sure your specimen is at the eucentric height and proper lens focus and adjustment are big factors. Diffraction was actually more precise in old 5 lens (1960s) TEMs compared to now. Split objective lenses with condensor minilenses that interact with the field below the specimen might make for high resolution images and small analysis spots but they cost the user in terms of diffraction precision. IMHO :-)
I have been asked the following question: ----------------------------------- I am looking for a peice to our old steroplotter it is 1940's model is a officine galieo made by the galieo company of italy. model number is smg10. do not know if you know this info but maybe you can stir in right direction . ---------------------------------- Does anyone have any information about this "galieo company of italy"? Sounds like this is a real museum piece.
Chuck
====================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ =====================================================
I have been asked the following question: ----------------------------------- I am looking for a peice to our old steroplotter it is 1940's model is a officine galieo made by the galieo company of italy. model number is smg10. do not know if you know this info but maybe you can stir in right direction . ---------------------------------- Does anyone have any information about this "galieo company of italy"? Sounds like this is a real museum piece.
Chuck
====================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ =====================================================
} Hello, } } I have stumbled across a reference to Hoffman modulation contrast (HMC). } Briefly, the article states that HMC allows 3-D like images. It is possible } to view specimens through plastic dishes (a limitation of Nomarski DIC ?) } and to view thick samples (a limitation of phase contrast). } } Can anyone tell me more about this technique please? } } Many thanks, } Felicity } } Felicity Lawrence
First, let me recommend Slayter & Slayter, "Light and Electron Microscopy". (Note: Nomarski=Differential Interference Contrast) Briefly simplified, with the attendent inaccuracies that implies: Hoffman Modulation works in a similar way to Nomarski, the major difference being that Nomarski is based on phase constrast between a specimen and a reference ray and Hoffman works by "amplitude modulation" between specimen and reference rays. Nomarski generates its specimen and reference rays with polarizers, Hoffman does not use polarized light. This means that Nomarski has troubles with optically active materials--specimens, plastic petri plates, etc.--whereas Hoffman does not. I've found that Nomarski seems to work better with thinner specimens (tho' I have used Nomarski with zooplankton), Hoffman with thicker, but I've read comments to the opposite. Hoffman does not have the troubles with convex/concave specimens that Phase Contrast does. Phil
&&& Illigitimi non carborundum &&&&&&&& Philip Oshel Microscopy Station A PO Box 5037 Champaign, IL 61825-5037 (217)244-3145 days (217)355-3145 evenings oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
I asked for information about Hoffman Modulation Contrast (HMC) and received the following replies :
1. I used Hoffmann several years ago to view dispersed polyethylene in PET sheeting for trays. I agree that HMC produces the illusion of 3-D, but to me the strong advantage was the ability to alter the "amount" of phase contrast which reduces the "halo" around particles. It seemed to work the best on multi-phase samples in which the difference between the refractive index of the phases were too high for phase contrast but too low for brightfield.
I use to cut my thin section samples by hand, so I do not have thickness data, but I found the thin samples gave better photomicrographs then thick samples. [F.K.]
2. If you have not already, you may wish to look at Hoffman's original article. [Hoffman (1977) J Microscopy 110, 205-22. [D.C.]
3. Try:
The Particle Atlas, ed. 2 Volume V pp. 1155-57
Ann Arbor Science Publishers, Inc. P.O. Box 1425 Ann Arbor, Michigan 48106
This reference is out of print (if memory serves me correctly) but many light microscopists have the reference. It was being sold at one time on CD-Rom.
It may be available still at the following:
Microscope Publications Ltd - Products Available from Microscope Publications Ltd. The Microscope Journal. Books in The Microscope Series. Other McRI Publications. Return to McRI Home.. --http://www.mcri.org/McRI_products.html [J.H.]
4. Have you tried Zeiss VAREL contrast? [D.F.]
5. We've used Hoffman illumination to good advantage with semi-thin (0.5 um ) sections of Spurr's resin embedded material (Sex. Plant. Reprod. 9:1-16). When we purchased the objective and condenser, the salesperson stressed that it was particularly useful for tissue culture observation with inverted optical scopes. It also gives nice images on wet mounts, at a considerably lower cost than Nomarski - although in my opinion, Nomarski is still superior for this purpose. The thickest material we looked at was probably ca. 10 um (squashes of Arabidopsis microsporocytes) and they looked good - but no plastic in that case. I did hear recently that Hoffman's patent is expiring and that the microscope companies that were originally not interested in his development are coming out with their "own" versions, so it's hard to tell what will happen to prices. I think we paid about $2000 U.S. for a 100X objective and condenser about 5 years ago. Maybe they're (Hoffman) having a sale now. [H.O.]
6. The system does indeed work and give DIC like images through plastic. It is not as good as DIC and from my experience, definitiey begins to degrade at higher magnification (the 40x lens is usable, but not great). I have only looked at cells, and not at thick tissue, so I don't know how suitable it is. Have a dealer bring in the system and try it out. It is not hard to add to an existing microscope. It is just a polarizer and a special lens with a polarizer element in it. I have them listed at 516-484-8882 Modulations Optics Inc. in New York. Hope this helps- Dave
Hoffmann modulation will allow you to view through plastic dishes. Neither Hoffmann modulation nor Differential Interference Contrast (Nomarski) give you 3D views...they are shadow casting methods depending on contrast generation due to differences in refractive index of different parts of the specimen. The reference for Hoffman Modulation is Hoffman, R. 1977. J. Microscp. 110:205. [N.A.]
7. First let me recommend Slayter & Slayter, "Light and Electron Microscopy". (Note: Nomarski = Differential Interference Contrast) Briefly simplified, with the attendent inaccuracies that implies : Hoffman Modulation works in a similar way to Nomarski, the major difference being that Nomarski is based on phase contrast between a specimen and a reference ray and Hoffman works by "amplitude modulation" between specimen and reference rays. Nomarski generates its specimen and reference rays with polarizers, Hoffman does not use polarized light. This means that Nomarski has troubles with optically active materials -- specimens, plastic petri plates, etc. -- whereas Hoffman does not. I've found that Nomarski seems to work better with thinner specimens (tho' I have used Nomarski with zooplankton), Hoffman with thicker, but I've read comments to the opposite. Hoffman does not have the troubles with convex/concave specimens that Phase Contrast does. [P.O.]
Felicity Lawrence
Analytical Electron Microscopy Facility Queensland University of Technology GPO Box 2434, Brisbane 4000 Australia
DOES ANYONE KNOW OF A USED ELECTRON MICROSCOPE VENDOR ON EAST COAST THAT BUYS OLD EM'S WE HAVE A JEOL 100B FOR SALE I HAVE SEEN CATALOGS FROM A BUSINESS CALLED "BID SERVICE" BUT I DIDNT SAVE ANY OF THEM
DOES ANYONE KNOW OF A USED ELECTRON MICROSCOPE VENDOR ON EAST COAST THAT BUYS OLD EM'S WE HAVE A JEOL 100B FOR SALE I HAVE SEEN CATALOGS FROM A BUSINESS CALLED "BID SERVICE" BUT I DIDNT SAVE ANY OF THEM
THANX IN ADVANCE
The "BID Service" phone number is (908) 775-8300.
Margaret E. Bisher
NEC Research Institute 4 Independence Way Princeton, NJ 08540. Tel.: (609) 951-2629 Fax: (609) 951-2496 e-mail: peggy-at-research.nj.nec.com
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} Hello, } } I have stumbled across a reference to Hoffman modulation contrast (HMC). } Briefly, the article states that HMC allows 3-D like images. It is possible } to view specimens through plastic dishes (a limitation of Nomarski DIC ?) } and to view thick samples (a limitation of phase contrast). } } Can anyone tell me more about this technique please? } } Many thanks, } Felicity } } Felicity Lawrence
First, let me recommend Slayter & Slayter, "Light and Electron Microscopy". (Note: Nomarski=Differential Interference Contrast) Briefly simplified, with the attendent inaccuracies that implies: Hoffman Modulation works in a similar way to Nomarski, the major difference being that Nomarski is based on phase constrast between a specimen and a reference ray and Hoffman works by "amplitude modulation" between specimen and reference rays. Nomarski generates its specimen and reference rays with polarizers, Hoffman does not use polarized light. This means that Nomarski has troubles with optically active materials--specimens, plastic petri plates, etc.--whereas Hoffman does not. I've found that Nomarski seems to work better with thinner specimens (tho' I have used Nomarski with zooplankton), Hoffman with thicker, but I've read comments to the opposite. Hoffman does not have the troubles with convex/concave specimens that Phase Contrast does. Phil
&&& Illigitimi non carborundum &&&&&&&& Philip Oshel Microscopy Station A PO Box 5037 Champaign, IL 61825-5037 (217)244-3145 days (217)355-3145 evenings oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
A short course in SEM for biologists will be held at the University of Florida March 10-13 and again May 5-8. This is for the beginner. Interested persons should see our WWW site for details.
www.biotech.ufl.edu/~emcl ******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
In response to your inquiry about Hoffman Modulation Contrast and the kind response by jeharper-at-amoco.com mentioning The Particle Atlas, I can provide the following information. The Particle Atlas Electronic Edition (PAE2) is available from the following suppliers:
MicroDataware P. O. Box 12755 Berkeley CA 94712 sshaffer-at-microdataware.com
McCrone Research Institute 2820 S. Michigan Avenue Chicago, IL 60616 ndaerr-at-mcri.org 1-312-842-7105 voice 1-312-842-1078 fax
McCrone Accessories and Components 850 Pasquinelli Drive Westmont, IL 60559 1-708-887-7100 voice 1-708-887-7764 fax
McCrone Scientific Ltd. McCrone House 155A Leighton Road London NW5 2RD UNITED KINGDOM 011 441 71 267-7199 voice 011 441 71 267-3383 fax
If and when I can get problems with my internet service provider ironed out I will put the MicroDataware web pages, with further information about the Particle Atlas Electronic Edition, back up at http://www.microdataware.com. However, for now this site is not active.
Best of luck with your HMC work. It is an excellent technique for contrast enhancement, yielding images similar in many ways to DIC.
Although it is a bit lengthy for a posting to a list server, I have included full text of the sections of the PAE2 dealing with HMC below. Information on this excellent technique is sufficiently scarce that I thought many of the readers of the list might find it, and the literature references, useful.
Steve Shaffer
Text on Hoffman Modulation Contrast from the Particle Atlas follows:
***************************************** Chapter 2: Techniques for Particle Characterization {snip} R. Hoffman Modulation Contrast
1. Introduction
A common problem in microscopy is object contrast. If the substage aperture has to be closed in order to see the object, the resolving power is severely reduced. Particle microscopists often attempt to maintain high resolution and at the same time enhance contrast by mounting their samples in Aroclor. This excellent mountant has a conveniently high refractive index, usually sufficiently different from the particles to permit use of a full condenser aperture and, hence, obtain maximum resolution.
Often, however, particle microscopists resort to a contrast enhancement technique such as darkfield, Zernike phase contrast or Nomarski differential interference contrast, DIC . Such methods increase the sensitivity of refractive index measurement besides making all particles more distinctly visible. On occasion, the mounting medium is not a matter of choice, and particle contrast cannot then be enhanced by choosing mountants of very different refractive index. Particles on a membrane filter, for example, are difficult to see unless the filter structure is cleared by immersing the filter and particles in a liquid matching the filter in refractive index. This is seldom the best liquid in which to study the particles. Counting asbestos fibers, usually chrysotile with indices near 1.55, is not easily done when the clearing liquid has to have an index of 1.51. Usually, in this situation, microscopists have turned to phase contrast to make the fibers easier to size and count.
2. Optics
A new contrast enhancement method with advantages over other methods has been developed by Hoffman and Gross since the publication of The Particle Atlas in 1973. Now termed Hoffman modulation contrast (HMC), this new system can be adapted easily to most microscopes. It enhances contrast without the halos characteristic of phase contrast and without apparent loss of resolution. The image, like DIC, is "three-dimensional" and permits optical sectioning of an object with little or no interference from detail above or below best focus. Also, like DIC, the effect is directional, and both detect phase gradients by converting opposite gradients into lighter or darker images compared to the background. The means by which this is accomplished is very different, however, for the two procedures. Nomarski, in his DIC system, uses a modified Wollaston prism to produce two sheared rays of light, separated by only about 1 micrometer, passing through the object, introduces interference with resulting darker borders on the other side. This produces a shadow effect and the three-dimensional appearance.
Hoffman and Gross, on the other hand, accomplish similar results by quite different optical principles. Figure 281 shows schematically the component required to convert a brightfield microscope for modulation contrast. A system of apertures below the substage condenser is conjugate with a matching system of apertures in the objective back focal plant. A full-aperture polarizing filter below the substage apertures permits variable contrast enhancement. It is essential that the substage apertures be fitted into a rotatable gliding mount in order to be able to accurately register their image with respect to a second set of apertures in the objective back focal (Fourier) plane.
3. Apparatus
The light source and microscope should be arranged for Kohler illumination. The aperture system below the substage condenser has about one-half its aperture covered (lengthwise) with a polarizing filter P1 (P2 is also a polarizer). The modulator has three regions differing in transmittance: D, 1%; G, 15%; and B, 100%. The 15% transmittance G region is not a polarizing filter.
In practice, the gray region (G) is displaced to one edge of the Fourier plane, and the dark region (D) lies outside the exit pupil of the objective. The bright region (B) then fills about 90% of the Fourier plane. Resolution of the system, approximating lambda/2NAobj. in spite of the restricted substage aperture, is maximized by off-setting the gray (G) region to the edge of the exit pupil. When the gray region is centered on the microscope optical axis, the resolution then approximates lambda/NAobj.. Removal of the aperture plate permits use of the HMC objective for ordinary brightfield study.
The system is so simple that many microscopists might consider making their own apertures; however, this same simplicity also makes professional conversion relatively low in cost. Note, however, that 3X to 100X objectives can be fitted for HMC, but each objective to be so used must be modified; each objective also requires its own substage aperture system.
4. Applications
HMC will be most useful for the biologist who always has contrast problems with tissue sections (Figures 282 and 286). From a strictly contrast point-of-view it will still be best to use mounting media having refractive indices very different from the object; however, when this is impossible, HMC will be very useful. Even when the indices of object and mount are quite different, HMC helps in the delineation of fine detail. Forensic scientists will find it helpful for semen examination and especially for species identification of spermatozoa by study of their morphological features. Banding of the sperm head as well as length and diameter measurement of the neck portions are much easier with HMC. HMC will also be helpful in enhancing contrast during refractive index measurement by the immersion method. In short, HMC will be useful as a replacement for phase contrast because of the better image, and for DIC because it is considerably cheaper.
Anyone interested in the theory of HMC can refer to the Hoffman and Gross papers, especially the 1977 paper in the Journal of Microscopy.
Hoffman, R., "The modulation contrast microscopy," Am. Lab. (1978).
Hoffman, R., and L. Gross, "Modulation contrast microscopy," Turtox News, 2-5, (1975).
Hoffman, R., and L. Gross, "The modulation contrast microscope," Nature 254, 586 (1975).
Hoffman, R., and L. Gross, "The modulation contrast microscope," Microscope 23, No. 4, 264 (1975).
Hoffman, R., and L. Gross, "The modulation contrast microscope," Appl. Opt. 14, 1169 (1975).
Hoffman, R., and L. Gross, "The modulation contrast microscope," J. Microsc. 110, 205 (1977).
Hoffman, R., and L. Gross, "The modulation contrast microscope," Chapter in Microstructural Science, Vol. IV, E. W. Filer et al., Eds., American Elsevier, New York, 1977, p. 287.
***************************************** End of excerpts from the Particle Atlas Electronic Edition
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I asked for information about Hoffman Modulation Contrast (HMC) and received the following replies :
1. I used Hoffmann several years ago to view dispersed polyethylene in PET sheeting for trays. I agree that HMC produces the illusion of 3-D, but to me the strong advantage was the ability to alter the "amount" of phase contrast which reduces the "halo" around particles. It seemed to work the best on multi-phase samples in which the difference between the refractive index of the phases were too high for phase contrast but too low for brightfield.
I use to cut my thin section samples by hand, so I do not have thickness data, but I found the thin samples gave better photomicrographs then thick samples. [F.K.]
2. If you have not already, you may wish to look at Hoffman's original article. [Hoffman (1977) J Microscopy 110, 205-22. [D.C.]
3. Try:
The Particle Atlas, ed. 2 Volume V pp. 1155-57
Ann Arbor Science Publishers, Inc. P.O. Box 1425 Ann Arbor, Michigan 48106
This reference is out of print (if memory serves me correctly) but many light microscopists have the reference. It was being sold at one time on CD-Rom.
It may be available still at the following:
Microscope Publications Ltd - Products Available from Microscope Publications Ltd. The Microscope Journal. Books in The Microscope Series. Other McRI Publications. Return to McRI Home.. --http://www.mcri.org/McRI_products.html [J.H.]
4. Have you tried Zeiss VAREL contrast? [D.F.]
5. We've used Hoffman illumination to good advantage with semi-thin (0.5 um ) sections of Spurr's resin embedded material (Sex. Plant. Reprod. 9:1-16). When we purchased the objective and condenser, the salesperson stressed that it was particularly useful for tissue culture observation with inverted optical scopes. It also gives nice images on wet mounts, at a considerably lower cost than Nomarski - although in my opinion, Nomarski is still superior for this purpose. The thickest material we looked at was probably ca. 10 um (squashes of Arabidopsis microsporocytes) and they looked good - but no plastic in that case. I did hear recently that Hoffman's patent is expiring and that the microscope companies that were originally not interested in his development are coming out with their "own" versions, so it's hard to tell what will happen to prices. I think we paid about $2000 U.S. for a 100X objective and condenser about 5 years ago. Maybe they're (Hoffman) having a sale now. [H.O.]
6. The system does indeed work and give DIC like images through plastic. It is not as good as DIC and from my experience, definitiey begins to degrade at higher magnification (the 40x lens is usable, but not great). I have only looked at cells, and not at thick tissue, so I don't know how suitable it is. Have a dealer bring in the system and try it out. It is not hard to add to an existing microscope. It is just a polarizer and a special lens with a polarizer element in it. I have them listed at 516-484-8882 Modulations Optics Inc. in New York. Hope this helps- Dave
Hoffmann modulation will allow you to view through plastic dishes. Neither Hoffmann modulation nor Differential Interference Contrast (Nomarski) give you 3D views...they are shadow casting methods depending on contrast generation due to differences in refractive index of different parts of the specimen. The reference for Hoffman Modulation is Hoffman, R. 1977. J. Microscp. 110:205. [N.A.]
7. First let me recommend Slayter & Slayter, "Light and Electron Microscopy". (Note: Nomarski = Differential Interference Contrast) Briefly simplified, with the attendent inaccuracies that implies : Hoffman Modulation works in a similar way to Nomarski, the major difference being that Nomarski is based on phase contrast between a specimen and a reference ray and Hoffman works by "amplitude modulation" between specimen and reference rays. Nomarski generates its specimen and reference rays with polarizers, Hoffman does not use polarized light. This means that Nomarski has troubles with optically active materials -- specimens, plastic petri plates, etc. -- whereas Hoffman does not. I've found that Nomarski seems to work better with thinner specimens (tho' I have used Nomarski with zooplankton), Hoffman with thicker, but I've read comments to the opposite. Hoffman does not have the troubles with convex/concave specimens that Phase Contrast does. [P.O.]
Felicity Lawrence
Analytical Electron Microscopy Facility Queensland University of Technology GPO Box 2434, Brisbane 4000 Australia
Of course the Bid Service buys old EM's. I have seen over 6 old SEMs in their catalog. Otherwise I think you can try CBI(Capovani Brother's Inc.). Their email and address are:
cbi-at-capovani.com ph: 518.346.8347 fax: 518.381.9578 704 Corporations Park Scotia, Ny 12302
Good luck.
Tong
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I can recommend two very good sources for information on the Poisson Distribution. 1. Statistical Analysis in Chemistry & Chemical Engr. by Bennett & Franklin, John Wiley, 1954, p115 (This is an older book, but one that is written very clearly and in a very practical orientation. It covers the basic statistical concepts plus a thorough treatment of several common experimental design schemes for analyzing data from complex multi-variable experiments.)
2. Data Reduction and Error Analysis for the Physical Sciences, by P. R. Bevington, McGraw-Hill, 1969, p.36. This is another very practical book that deals particularly with the counting of photons and particles. It also contains algorithms for (in FORTRAN) for a wide variety of data analysis processes (deconvoluting spectral data, smoothing, peak area measurement, etc).
wow this is service thanx to all the replys for bidservice info happened to check the web and the right place to start is at Microworld resource net at http://www.mwrn.com/product/microscope/used.htm pointed to at least 6-7 companies
sorry joe at ddk about the SHOUT ing didnt know caps lock was so loud
I have just setup a microscope system for use with an existing picosecond pulsed laser to perform time-resolved fluorescence measurements. Currently we are using an inverted Zeiss Axiovert IM-35 and a photon-counting photomultiplier to perform spot-measurements. We are considering an upgrade to the present system to include a confocal scanning attachment in order to be able to acquire images as well as fluorescence decays. The laser we are considering is a Ti-sapphire with picosecond/femtosecond pulse capabilities, so that 2-photon excitation imaging would also be possible. Could anybody recommend an attachment capable of adding confocal capabilities to our Zeiss microscope? Any comments, suggestions, warnings, etc. would be greatly appreciated.
Thank you in advance.
Massimo Sassaroli, D.Sc. Dept. of Physiology & Biophysics Box 1218 Mount Sinai School of Medicine 1 Gustave L. Levy Pl. New York, NY 10029-6574
We have a Hitachi S-800 FESEM from the pre-digital days. We have been saving up our pennies to purchase either the Hitachi PCI system (many pennies) or the GW Electronics Printerface (fewer pennies). We got as far as getting a great new PC before we ran out of pennies. However, we did purchase with the FESEM a S-5010 TV Scanning Device, which basically converts Hitachi's propriatary signal to NTSC, primarily for output to a VCR. This device does not give the same image as on the CRT, and one has to use specific accelerating voltages and working distances and read the magnification off a chart. And the magnification range is very limited, making it useless for our high mag-high res work. However, I suspect it could be used as an interim device for grabbing images with the PC. I have snaked a long cable from it to the Mac and used the Power PC's video board to get (rather lousy) images. I would now like to get a video board for the new Pentium and do the same (or better). I don't know video boards. I am looking for advice! If I could get some digital images for the price of a video board, I would be thrilled! I promise disgusting weather reports to all who respond...
Thank you all for being such a friendly and helpful group!
Aloha, Tina
New Stuff! http://www.pbrc.hawaii.edu/bemf/microangelo Simple SEM animations at- http://pbrc.hawaii.edu/bemf/microangelo/gif_animations.html **************************************************************************** * 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* ****************************************************************************
Hi all, I'll keep this short. I've been asked to post concerning the availability of two free TEM's here at the University of Iowa. There is a Siemans 101 and a Phillips 201. To the best of my knowledge, they are both in operating condition. For further information, contact Kenneth Moore at {kenneth-moore-at-uiowa.edu} or phone 319-335-8143. Thank you.
A month ago I asked for comments and experiences relating to allergic reactions to the use of acrylate embedding media. To date, I have received the following responses. I thank you all for your input.
Have attached the file of responses as direct e-mail seems to be difficult or impossible with my internet connector...
I just began getting replies to my request for advice on video boards for digital capture from my analog FESEM. Yes, I DO want a high-quality system - eventually. But until the University recovers from fiscal insults, I am talking about a SUPER-CHEAP way out. I can get a (not very nice) NTSC signal from a Hitachi device I already have, so I only want right now a card to stick in the PC (which is in the same room) for something like $200 to $400. Like what I can pay for out of my own pocket. I am still very interested in people's experiences with full systems, since that is where we eventually want to go, but I'm only trying to kludge something together for the moment! I will happily take all opinions on both "real" digital acquisition systems and "kludge" solutions!
Yeah, I know it's silly to count pennies when I have a $200K 'scope, but right now even Kimwipes(tm) are at a premium!
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
I just began getting replies to my request for advice on video boards for digital capture from my analog FESEM. Yes, I DO want a high-quality system - eventually. But until the University recovers from fiscal insults, I am talking about a SUPER-CHEAP way out. I can get a (not very nice) NTSC signal from a Hitachi device I already have, so I only want right now a card to stick in the PC (which is in the same room) for something like $200 to $400. Like what I can pay for out of my own pocket. I am still very interested in people's experiences with full systems, since that is where we eventually want to go, but I'm only trying to kludge something together for the moment! I will happily take all opinions on both "real" digital acquisition systems and "kludge" solutions!
Yeah, I know it's silly to count pennies when I have a $200K 'scope, but right now even Kimwipes(tm) are at a premium!
**************************************************************************** * 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* ****************************************************************************
Division for Microscopy and Microanalysis Department of Physics Chalmers University of Technology S-412 96 G=F6teborg, Sweden
Two PhD student positions are open in Materials Science/Engineering with the following contents.
1. CONTACT AND ADHESION BETWEEN SMALL PARTICLES. Contact and spontaneous adhesion between small particles (10-100 nm) will be studied with analytical and high resolution electron microscopy. The spontaneous adhesion between small particles depends on surface forces. The accompanying stress fields, which are visible in the electron microscope, give a unique possibility in detail to study the elastic and plastic deformations which are associated with contact. This effect is very importannt within areas such a friction, wear, fretting, electrical contacts and handling of fine powders.
2. NANOSTRUCTURED MATERIALS. GRAIN BOUNDARIES AND MECHANICAL PROPERTIES. Nanostructured material exhibit many new and interesting properties due to their extremely fine-grained structure ( { 100 nm). The reduction in grain size leads to unique mechanical, physical and chemical properties compared with conventional materials with the same composition. The mechanism behind the formation and the stability of different nanostructures and the resulting properties are however poorly understood. The idea with the current program is to establish a systematic knowledge about the grain boundary zone in nanostructured materials and their influence on the mechanical properties. A combination of advanced electron microscope and analytical methods will be used.
The two projects lie in the boundary area between physics/materials science. In both projects a Philips CM 200 FEG with Gatan imaging filter, slow scan CCD-camera, energy dispersive X-ray analysis and PEELS will be used.
Qualifications: Master of Science or equivalent within physics/materials science. A knowledge of electron microscopy is estimated but is not absolutely necessary. A vivid interest for mathematics is appreciated.
Further information: Professor Anders Tholen,=20 Division for Microscopy and Microanalysis Department of Physics Chalmers University of Technology SS-412 96 Goteborg, Sweden
Tina, I am a microscopist in the research laboratories of Eastman Chemical Company. Although I haven't had any personal experience with a product called "Snappy", which is an adapter for NTSC image capture on a PC, it might be just what you are looking for. In sells for under $200. A friend of mine at Clinch Valley College is using one on an AMRAY SEM and is quite pleased.
I just checked out your BEMF pages and they are quite nice! One note, though. The net address you listed in your note (see copy below) to the IT list-server left out the "bemf" part.
} Mahalo, } Tina } } http://www.pbrc.hawaii.edu/microangelo } http://www.pbrc.hawaii.edu/microangelo/gif_animations.html }
The address I used was
http://www.pbrc.hawaii.edu/bemf/microangelo/
Dr. Dennis B. Barr | 333 DDDDD 333 DDDDD Eastman Chemical Company | 3 3 D D 3 3 D D Microscopy and Morphology | 3 D D 3 D D Research Laboratory | 33 D D 33 D D P.O. Box 1972 | 3 D D 3 D D Kingsport, TN 37662-5150 | 3 3 D D 3 3 D D | 333 DDDDD 333 DDDDD Voice: 423/229-2188 | E-mail: dennbarr-at-eastman.com | IMAGE IMAGE FAX: 423/229-4558 |----------------------------------------------------- (Cross your eyes to see the 3 dimensional image above)
About a month ago I asked for experiences with allergy experiences when using acrylate embedding media. I received the responses below. I will be sending several editions as there are many and I have had difficulties with my e-mail server...too big a gulp, I guess...
1) I used Lowicryl for a brief time, while waiting for funding for a cryokit. I developed a pretty good contact dermatitis/allergic reaction within a few uses, precluding me using it. Also I found the smell very allergenic (runny nose etc) Luckily I got the cryokit, and was able to return to a NON allergenic system (sugar!) very soon. Since then I have been convinced that the acrylics are bad news..... Good luck with your investigations.
Simon
2) Their was a program on NOVA that concerned itself with Sick Building Syndrome. It covered several people in a hospital environment that aquired hyper-allergies from the gloves and chemicals used. You might want to check the references that the film used... It was a good..... Check with WGBH in Boston (PBS Station) for more information on the film.....
I have seen several technicians and operators become sick from the fumes from roughing pumps and from Freon that is widely used.. Anyone of us could be next with the wide variety of chemical we use and the increasing amount of contact we have....
Walter Protheroe
3) Over the years, I have picked up some sense of the way these different acrylics seem to act on different persons.
The Lowicryl resins seem to be the worst, and LRWhite (and I thought up until now, also Unicryl) the least reactive. Technovit I have not had any knowledge about.
Chuck
Regards, Don Cox ******************************************************** Donald P. Cox, Ph.D., M.B.A. GOLDMARK BIOLOGICALS/D.P. COX ASSOCIATES 437 Lock Street, Phillipsburg, NJ 08865-2764 (908) 859-2631 - - (908) 859-2875-FAX E-Mail: goldmrkr-at-fast.net/goldmarker-at-aol.com Web Page: http://members.aol.com/goldmarker
~~~"Goldmarking is everlasting probing!"~~~ ********************************************************
About a month ago I asked for experiences with allergy experiences when using acrylate embedding media. This is the second series..
5) My predecessor here had a lot of problems using watyer soluble Durcupan resin. He manifested what we now realise was contact dermatitis, with skin blemishing, cracking and peeling. He was also sensitive to the cured blocks, at least certainly to their dust (we used to cure in ice cube trays and then cut specimens out with a hacksaw for mounting prior to ultramicrotomy as routine).
With best wishes - Keith Ryan
6) Don, I realize that you asked about acrylic plastics, but just as a note: my former supervisor in a clinical EM lab had a strong contact allergy to uncured epoxy plastics. We had to be careful not to leave residue where she could come in contact with it, she'd break out in blisters in about 15 min.
Doug
7) have you got this reference? It seems to be a key one referring to several others:
Tobler, M. and Freiburghaus (1990); Occupational risks of (meth)acrylate compounds in embedding media for electron microscopy Journal of Microscopy 160: 291-298
Good luck in your search.
Malcolm Haswell
8) Don, I don't know what to tell you except that this allergy is common, I am severly allergic to Lowicryl, and have been told that if I'm exposed to it again on my hands, I may lose the use of my hands. It took me 6 months to get the feeling back in my finger tips, and my fingers were so bad they were purple black. Also, the fumes from methacrylates etc is of no help to my chronic asthma.
I usually run into someone with the same problem in the Histology / EM field where ever I go.
We ordered the only gloves we found impermiable, H-4 Gloves from Monsanto, but they are very ackward to work in.
Lou Ann
----------------------
Regards, Don Cox ******************************************************** Donald P. Cox, Ph.D., M.B.A. GOLDMARK BIOLOGICALS/D.P. COX ASSOCIATES 437 Lock Street, Phillipsburg, NJ 08865-2764 (908) 859-2631 - - (908) 859-2875-FAX E-Mail: goldmrkr-at-fast.net/goldmarker-at-aol.com Web Page: http://members.aol.com/goldmarker
~~~"Goldmarking is everlasting probing!"~~~ ********************************************************
I would like to obtain an old Balzers EVM 052/052 Electron Beam Gun power supply. I'm also interested in any unused or unwanted Balzers Freeze Fracture systems or parts.
Dear Tina, I'm afraid that the best you will be able to do is a lousy image, no matter what video board you get. The rapid scan image on any SEM is very poor, that is why slow scan capture is used for digital imaging. You wrote: } } We have a Hitachi S-800 FESEM from the pre-digital days. We have been } saving up our pennies to purchase either the Hitachi PCI system (many } pennies) or the GW Electronics Printerface (fewer pennies). We got as far } as getting a great new PC before we ran out of pennies. However, we did } purchase with the FESEM a S-5010 TV Scanning Device, which basically } converts Hitachi's propriatary signal to NTSC, primarily for output to a } VCR. This device does not give the same image as on the CRT, and one has } to use specific accelerating voltages and working distances and read the } magnification off a chart. And the magnification range is very limited, } making it useless for our high mag-high res work. However, I suspect it } could be used as an interim device for grabbing images with the PC. I } have snaked a long cable from it to the Mac and used the Power PC's video } board to get (rather lousy) images. I would now like to get a video board } for the new Pentium and do the same (or better). I don't know video } boards. I am looking for advice! If I could get some digital images for } the price of a video board, I would be thrilled! I promise disgusting } weather reports to all who respond... } } Thank you all for being such a friendly and helpful group!
Regards, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
Colleagues - Continuing.... You'll never ask again, I'll bet...!
11) } i do not recall her name off hand, but if you contact rmc,inc } tucson,arizona,(they probably have a web page) and ask who was their } biological instructor at their ultra microtomy conference this year in } oct . I specifically remember, and i was surprised that she repeatedly } cautioned the group about similar severe allergy-proned } individual-reactions being rather common. I believe she also may be } linked to this newsgroup and may reply directly.
I'm that instructor. I don't have any specific medical information; just the sort of word-of-mouth stories that are appearing here (which I'm saving for next year's Materials Microtomy course!). I'd appreciate comments from knowledgable MDs. I know that I read a paper some years ago about similar reactions to the acrylic resins used to cement metal joints into bone in orthopaedic surgery...
Caroline Schooley
12) Hi: I myself have through the years developed a reaction to acrylates. My field is polymers and I am frequently in contact with acrylates. The monomer is the problem not the polymer. Wear gloves and always work in the hood. If you do get exposed wash or shower as soon as possible. The reaction is cumulative and becomes worse with each exposure. Good luck Olga L. Shaffer
13) Here at IBM, there is a fairly extensive Industrial Safety and Hygiene program. In our training classes, it was explained to us that epoxies, etc., are "sensitizers" as are Poison Ivy, Poison Oak, and Poison Sumac. Some people may have a reaction on their first exposure. Others may have a reaction after repeated exposures have "sensitized" them. The reactions will most likely increase in sevearity with each additional exposure. There are a few that never show a reaction. I know people that were never affected by Poison Ivy when they were children, but had severe reactions when exposed as an adult.
Precautions during use (what we are required to do): * The resins and hardeners are stored in a vented cabinet. * Weighing, mixing, and sitting to cure are done in a vented hood. *** Protective Appearal *** *** Nitrile Gloves - eg. Nitrilite by Ansell Edmont Note: Latex gloves were not accepted * Goggles * Plastic Chemical Apron
Hi again,
(Please ignore my spelling typo's, it was late last night.) I meant to mention that the Nitrilite gloves are lightweight surgical type gloves, and very easy to work in.
Hope this helps.
Darrell Miles ******************************************************** Donald P. Cox, Ph.D., M.B.A. GOLDMARK BIOLOGICALS/D.P. COX ASSOCIATES 437 Lock Street, Phillipsburg, NJ 08865-2764 (908) 859-2631 - - (908) 859-2875-FAX E-Mail: goldmrkr-at-fast.net/goldmarker-at-aol.com Web Page: http://members.aol.com/goldmarker
~~~"Goldmarking is everlasting probing!"~~~ ********************************************************
About a month ago I asked for experiences with allergy experiences when using acrylate embedding media. This is the third series..
9) I have personal experience with methyl methacrylate contact allergies. Years ago, when I was taught how to embed and section JB-4 (from Polyscienses), I was told that the solutions could cause a poison ivy-like reaction if I got it on my skin. My teacher, however chose not to were gloves, so neither did I. After a couple of months of continuous use, I developed the worst reaction that you could imagine. The fingers that had been in contact with the liquid form of the acrylic started to itch and burn, deep inside the flesh. Next, my fingers swelled to the size of a meaty hotdog and itched like crazy. Any attempt to even touch then sent sharp pains through my hand. I took antihistamines as soon as the reaction started and applied ointment for two days before the reaction was over. After that episode, I accidently touched the solid form once or twice and scrubbed my hands immediately and at length And took some more antihistamines. Still, I got a minor reaction that lasted a day each time. Now, about 10 years later, I still use JB-4 for alot of our samples, but I Always were gloves, and glasses when I'm arround it and I always wipe down work surfaces when I'm finished for the day. I have not had another even minor reaction for at least the last 5 years and never one as bad as the first. The reactions to these acrylics can be quite painfull, but many of the other embedding medias available are carcinogenic, with no early warnings. At least with this material, I know immediately when I've gotten too careless with my safety precautions.
Karen Pawlowski
10) i do not recall her name off hand, but if you contact rmc,inc tucson,arizona,(they probably have a web page) and ask who was their biological instructor at their ultra microtomy conference this year in oct . I specifically remember, and i was surprised that she repeatedly cautioned the group about similar severe allergy-proned individual-reactions being rather common. I believe she also may be linked to this newsgroup and may reply directly. i believe she was retired and now consulting from her home in the Stanford area. when i can get to my records i'll try to remember to recover her name and send her e-mail address. i'm quite bad with names.
Charles J. Day
---------
Regards, Don Cox ******************************************************** Donald P. Cox, Ph.D., M.B.A. GOLDMARK BIOLOGICALS/D.P. COX ASSOCIATES 437 Lock Street, Phillipsburg, NJ 08865-2764 (908) 859-2631 - - (908) 859-2875-FAX E-Mail: goldmrkr-at-fast.net/goldmarker-at-aol.com Web Page: http://members.aol.com/goldmarker
~~~"Goldmarking is everlasting probing!"~~~ ********************************************************
I had difficulty forwarding a very helpful response from Stephen Griffiths and if it does not go this time I will re-write it....
5) Hi Don
I have certainly had contact dermatitis from use of Lowicryl K4M. Not an allergic reaction though.
I discovered, after using K4M with latex or plastic gloves for several years that Lowicryl penetrates within a few minutes. There are a couple of papers, I've no doubt you are familiar with them, which dealt with the subject. They were primarily concerned with "pushing" 4H Gloves but still interesting for all that.
I have the papers somewhere but couldn't put my hands to them, so I found the Refs from BIDS, which follow (I think this is technically a breach of their copyright, but it saves me digging around in boxes for an hour)
**************************************************************************** **************** 1) TI: A GLOVE WITH EXCEPTIONAL PROTECTIVE FEATURES MINIMIZES THE RISKS OF WORKING WITH HAZARDOUS CHEMICALS AU: TOBLER_M, FREIBURGHAUS_AU NA: UNIV HOSP ZURICH,DEPT INTERNAL MED,H LAB 8,CH-8091 ZURICH,SWITZERLAND JN: CONTACT DERMATITIS, 1992, Vol.26, No.5, pp.299-303
2) TI: EXCEPTIONAL PROTECTIVE POWER OF THE 4H GLOVE DEFEATS OCCUPATIONAL RISKS IN ELECTRON-MICROSCOPY AU: TOBLER_M, FREIBURGHAUS_AU NA: UNIV SPITAL ZURICH,DEPT INNERE MED,H LAB 8,CH-8091 ZURICH,SWITZERLAND JN: JOURNAL OF MICROSCOPY-OXFORD, 1991, Vol.163, No.JUL, pp.RP 1-RP 2
There is this problem, that using gloves gives a sense of false security. But the above papers show that penetration occurs very rapidly with ordinary gloves.
My problem may have arisen from handling improperly polymerised blocks without gloves during sectioning. The pattern of the dermatitis would seem to be consistent with that.
I tried 4H gloves. They were doubtless very effective, but difficult if you were handling small blocks and BEEM capsules. 4H gloves are a bit like wearing Aluminium foil. Not exactly sensitive!
If your customer absolutely HAS to use acrylics then s/he may find 4H gloves of some use. Though if it is a real full blown reaction the best thing they can do is avoid it at all costs, 4H gloves or no 4H gloves.
Eczema or contact dermatitis seems to be the favourite reaction to acrylics. I don't have to use them at the moment and the condition of my fingers has improved after three years. However once you get this sort of thing it never really goes away and can be triggered by agents other than the original one.
This can have its benefits occasionally as it gets me out of the washing up at home when I am having a recurrence of the condition. ;)
Regards Stephen Griffiths
AND FINALLY..... 14) It would be nice if you'd summarize and post the information you might receive on this subject. Safety in the laboratory is so important and people don't always realize (or remember) that. Thank you very much in advance. Adriana
-------------
Whew! That's over...!
Regards, Don ******************************************************** Donald P. Cox, Ph.D., M.B.A. GOLDMARK BIOLOGICALS/D.P. COX ASSOCIATES 437 Lock Street, Phillipsburg, NJ 08865-2764 (908) 859-2631 - - (908) 859-2875-FAX E-Mail: goldmrkr-at-fast.net/goldmarker-at-aol.com Web Page: http://members.aol.com/goldmarker
~~~"Goldmarking is everlasting probing!"~~~ ********************************************************
For your short term problem, get a "Snappy" frame grabber. Cheap but quality per $ spent is very very good. It's also very simple to use, just plug it into your paralell port.
Check out this site: http://www.zdnet.com/pccomp/sneakpeeks/snpk1096/snappy.html
That price for a dual Pentium system seems a bit high. The machine I'm using now, I put together as a motherboard and hard drive upgrade to a Gateway 486. It is a dual Pentium (133) PCI-EISA board with 512 K pipelined burst cache and 32 megs of RAM. Nothing around here can touch it in terms of speed and performance. Price of the upgrade (which I did myself): about $3200, including Windows NT to support the dual processor.
-=W.L. Steffens=- Department of Veterinary Pathology College of Veterinary Medicine University of Georgia
I use a commercial program from a company in your home town of Bochum, Germany. See their web site at www.tech-inter.com phone +49 234 30 96 96. The program's name is PicDoc. best regards mark
Mark W. Lund, PhD Director } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. ************************************************* Orem UT 84057 **"Soft x-rays in the 21st Century" conference ** 801-225-0930 ** 8-11 January 1997, Midway Utah ** FAX 801-221-1121 ** http://volta.byu.edu/xray/info.html ** lundm-at-xray.byu.edu *************************************************
"Let me commend a great truth to you which has been one of the supports of my life: 'The Gods send threads for a web begun.' Andrew Carnegie
Back in December I asked what X-Y-Z stage controllers were out there for an inverted scope & mac compatable.
Here's a summary of the replies...
Signal Analytics Vienna, VA 703-281-3277 $13k ***************************** New England Affiliated Technologies 620 Essex St. Lawrence, MA 01841 Tel 508-685-4900 or 800-227-1066 Fax 508-688-8027 $5k ***************************** Prior Scientific, 800-877-2234 $13k ***************************** thanks to all who helped.
I am interested in fluorescence imaging of single cells. However, these = cells will be in suspension. The medium will be quiescent and I want to = disturb the cells as little as possible. To that end, does there exist = a dye or a technique whereby the dye does not need to be rinsed from the = medium? =20
Are there other possibilities, such as a fluorescent dye that is = effective more than a couple hours after the cells have taken it up, or = slow release of the dye from beads, etc?
I will most likely be doing calcium measurements on mammalian bone or = smooth muscle cells.
Hello, I was wondering if anyone read the article about SGI's O2 system in the Jan 97 issue of BYTE. It compares the single processor O2 to a dual 200MHz Pentium Pro. The price of the tested SGI was $8342 while the price of the tested Pentium was $11,432. The article also quotes that O2 starting prices are below $6000.
The O2 model they wrote about had 128Mb of RAM, 2 2-Gb hard drives, and a 180MHz clock. The dual Pentium model also had an extra 16Mb of something called "Intense 3D".
The artice compared OpenGl performance. The dual Pentium performance was only slightly better (for $4000 more). I wonder about floating point operations though. I would be surprised if the dual Pentium could perform more quickly in that area, based on the Pentium Pro and R5000 (O2's CPU) floating point specifications I've seen/heard.
I'm wondering what the microscopy community's views are on the PC vs. UNIX (SGI specifically) issue. Or does anyone have anymore information on this topic?
Personally, I was really surprised by the article in BYTE even though it's only an OpenGl comparison. I had heard about how the dual Pentium Pro's were going to blow the SGI's out of the water and at a cheaper price. The hardware alone that is being bundled with this SGI was a shock given past SGI prices I've seen (in an academic setting).
We want to start with image analyzation andhave no idea about it, yet. What we have is a Zeiss Axioplan microscope with a Sony CCD camera (DXC-101P) attached to it. It has a resolution of 320x350 lines. There is also a photo camera for slides and negatives.
We need to measure areas in light microscopical sliceseither manually and automatically by their grey values. An option for 3D reconstruction would be nice, too.
My questions are now: 1) What further equipment do we need (frame grabber? better (digital?) camera? PC, MAC, workstation?)
2) Which software do we need? (what makes the difference between shareware (e.g. NIH Image) and expensive programmes to buy for some k$?
3) Is it worth to buy a commercial available complete system?
These questions may be very basic so possibly there is a site to look for FAQ like this. Which newsgroups cover this topic?
Any suggestions are welcome.
Hans-Martin
************************************************************** Hans-Martin Vaihinger Ruhr-University of Bochum Comparative Endocrinology Research Section Building ND 5/37 44780 Bochum GERMANY ********************************************************* phone ++49 234 700 4329 fax ++49 234 709 4551 email Hans-Martin.Vaihinger-at-RZ.Ruhr-Uni-Bochum.de
There was also a report about a no-nonsense new Motorola chip, the X704, which would be a 530 MHz puppy to be fitted into the next generation MACS. Maybe we can go back to user-friendly systems with excellent performance... Windows just doesn't quite cut it yet.
} X704, which would be a 530 MHz puppy to be fitted into the next What would the cost of a system with that kind of chip, the data bus of an SGI, and the rest of the accessories be like?
} That price for a dual Pentium system seems a bit high. The machine } I'm using now, I put together as a motherboard and hard drive upgrade } to a Gateway 486. It is a dual Pentium (133) PCI-EISA board with 512
Are those Pentium Pro chips? That's what they were looking at in the article I read.
--IMA.Boundary.541876258 Content-Type: text/plain; charset=US-ASCII Content-Transfer-Encoding: 7bit Content-Description: cc:Mail note part
I would like to thank Don for posting replies to his query about sensitivity to acrylics. No, Don, it shouldn't be "Whew! That's over...!" (I know what you mean), for anyone in the microscopy field who is exposed to chemicals, radiation, and/or any other hazard - it should be just the beginning!. I would ask everyone to consider that we do no know what the long term effects of exposure to many individual chemicals or combinations of chemicals can have on your health. Are you willing to risk a terminal illness when it is relatively easy for you can work in a fume hood and wear personal protective gear? I would hope not!
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I had difficulty forwarding a very helpful response from Stephen Griffiths and if it does not go this time I will re-write it....
5) Hi Don
I have certainly had contact dermatitis from use of Lowicryl K4M. Not an allergic reaction though.
I discovered, after using K4M with latex or plastic gloves for several years that Lowicryl penetrates within a few minutes. There are a couple of papers, I've no doubt you are familiar with them, which dealt with the subject. They were primarily concerned with "pushing" 4H Gloves but still interesting for all that.
I have the papers somewhere but couldn't put my hands to them, so I found the Refs from BIDS, which follow (I think this is technically a breach of their copyright, but it saves me digging around in boxes for an hour)
**************************************************************************** **************** 1) TI: A GLOVE WITH EXCEPTIONAL PROTECTIVE FEATURES MINIMIZES THE RISKS OF WORKING WITH HAZARDOUS CHEMICALS AU: TOBLER_M, FREIBURGHAUS_AU NA: UNIV HOSP ZURICH,DEPT INTERNAL MED,H LAB 8,CH-8091 ZURICH,SWITZERLAND JN: CONTACT DERMATITIS, 1992, Vol.26, No.5, pp.299-303
2) TI: EXCEPTIONAL PROTECTIVE POWER OF THE 4H GLOVE DEFEATS OCCUPATIONAL RISKS IN ELECTRON-MICROSCOPY AU: TOBLER_M, FREIBURGHAUS_AU NA: UNIV SPITAL ZURICH,DEPT INNERE MED,H LAB 8,CH-8091 ZURICH,SWITZERLAND JN: JOURNAL OF MICROSCOPY-OXFORD, 1991, Vol.163, No.JUL, pp.RP 1-RP 2
There is this problem, that using gloves gives a sense of false security. But the above papers show that penetration occurs very rapidly with ordinary gloves.
My problem may have arisen from handling improperly polymerised blocks without gloves during sectioning. The pattern of the dermatitis would seem to be consistent with that.
I tried 4H gloves. They were doubtless very effective, but difficult if you were handling small blocks and BEEM capsules. 4H gloves are a bit like wearing Aluminium foil. Not exactly sensitive!
If your customer absolutely HAS to use acrylics then s/he may find 4H gloves of some use. Though if it is a real full blown reaction the best thing they can do is avoid it at all costs, 4H gloves or no 4H gloves.
Eczema or contact dermatitis seems to be the favourite reaction to acrylics. I don't have to use them at the moment and the condition of my fingers has improved after three years. However once you get this sort of thing it never really goes away and can be triggered by agents other than the original one.
This can have its benefits occasionally as it gets me out of the washing up at home when I am having a recurrence of the condition. ;)
Regards Stephen Griffiths
AND FINALLY..... 14) It would be nice if you'd summarize and post the information you might receive on this subject. Safety in the laboratory is so important and people don't always realize (or remember) that. Thank you very much in advance. Adriana
-------------
Whew! That's over...!
Regards, Don ******************************************************** Donald P. Cox, Ph.D., M.B.A. GOLDMARK BIOLOGICALS/D.P. COX ASSOCIATES 437 Lock Street, Phillipsburg, NJ 08865-2764 (908) 859-2631 - - (908) 859-2875-FAX E-Mail: goldmrkr-at-fast.net/goldmarker-at-aol.com Web Page: http://members.aol.com/goldmarker
~~~"Goldmarking is everlasting probing!"~~~ ********************************************************
Received: from mpg.mcgawpark.baxter.com (159.198.180.50) by ccmailgw.mcgawpark.baxter.com with SMTP (IMA Internet Exchange 2.03 (Beta 5) Enterprise) id 0004CCC3; Tue, 7 Jan 97 15:15:51 -0600 Received: from sparc5.microscopy.com by mpg.mcgawpark.baxter.com id aa11752; 7 Jan 97 15:14 CST Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id KAA10359 for dist-Microscopy; Tue, 7 Jan 1997 10:51:30 -0600 Received: from fast.net (po.fast.net [198.69.204.10]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id KAA10356 for {microscopy-at-Sparc5.microscopy.com} ; Tue, 7 Jan 1997 10:51:26 -0600 Received: from lizard by fast.net with smtp (Smail3.1.29.1 #2) id m0vher2-0004KIC; Tue, 7 Jan 97 11:58 EST Message-Id: {m0vher2-0004KIC-at-fast.net}
Seeing the recent posting re Balzers Electron Beam Gun power supply prompts me to ask this: I run a 25-year-old JEOL microprobe, with very little chance of ever getting funds to buy a new replacement. One of these days the HV and gun filament supply (which uses vacuum tubes, including, for all the old-timers, a type 807!!!) is going to up and die, possibly by catastrophe within the tank. Are there available 3rd-party supplies which I could just graft in (maybe necessitating the fabrication of a custom cable) in that event? Is that what the aforementioned Balzers thingummy is? Anybody got an address for them or for anyone else who makes this sort of thing?
Happy New Year
Ritchie
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
Thanks to all who responded to my request about image capture systems. I have included a sanitized version of all the responses (I removed most vendors names and the like). I have all the original posts so email me if anything catches your eye and you need more particulars.
Bob Wise UW-Oshkosh
******************
I maintain an archive of most of the biologically relavant postings to this list. You are correct that this has been a hot topic in the last few months and I have archive it at our web site. Go to the URL listed at the end of this message and click on the "Tips & Tricks" link. Within there you will find a section on "Computers" Look at the section titles "Acquiring Digital Images". Let me know and I can get you the info in some other way if you need. http://www.biotech.ufl.edu/~emcl/
***********
We also have the Hitachi/Quartz PCI imaging system. I recall it being pricey, but we have been satisfied with it. It takes some learning, but is rather easy to use. It does annotation, some measurements, databases of sessions, etc. And they allow (even encourage) us to freely distribute their viewer software. Even though it is of limited capability, it still does quite a bit.
We do most of our output on a HP 4M Laserjet. At times we wish for better grayscale rendition. But the PCI allows us to send our images back to the scope's Polaroid camera if we need to.
***********
We use ImageSlave capture boards which we have fitted to each of our 6 microscopes. They fit in a standard slot in any old PC. They can run in Windows, but we run ours in DOS. Its so simple. Every time you push the PHOTO button, the image going to the photo display gets digitised ( putting in a film is an option, now uncommon with us). All you need to do is give a filename and the picture is saved to disk. The images are 1024x 1024 x 256 which we find perfectly good enough for our work. They are US $5,000 including software.
***********
One system that I think you ought to take a close look at is SEMICAPS. In 1990, I had one interfaced to my (then) Cambridge S200 SEM and I was very pleased with the results. At that time, I was also quite impressed by the customer service elicited by SEMICAPS. I have since moved this system to my light microscope for image analysis purposes. The SEM now has a new Link ISIS EDS system which has its own imaging capability. I am sure that the new SEMICAPS systems are even better, and I would hope that the customer service is still world class.
**************
The PCI system we sell is easily installed and very easy to learn how to use.
**************
We are looking into representing a company that makes a powerful, yet relatively inexpensive PC-based active system that includes the control board and software, and we can certainly configure it with a PC. We have access to a passive acq. system also, but I do not yet have much detail on this one. If you are interested, please feel free to contact me. We also represent a company who makes a versatile SW product for digital image archive and databasing, thumbnail viewer, etc. I'd be happy to discuss these and our new Image Analysis SW, Visilog in more detail, if you are interested.
**************
I am in a similar position of trying to set up an image capture system from our SEM also. I have just taken a work transfer, and have the challenge of setting up an EM/imaging facility where there was none before. Lots of fun, and something new for me, although I have spent almost 20 years doing EM, I have never actually been responsible for setting up or running the lab!!
I am starting to investigate options to get this system set up. It seems that we already have an image analysis software package (SigmaScan Pro from Jandel) and a fancy Pentium to run it on. In terms of the actual capture system for the SEM, I have heard from a number of my colleagues that the Hitachi PCI system is the best one out there, in their opinion. It is an entire system, including the computer, which can be augmented to be used with LM and TEM (for more money of course). It is expensive ($12.5K CAN) but is apparently worth every penny (we couldn't afford it). Another company in the US (4pi) sells a PCI system for quite a bit less money, but you have to supply your own Pentium or Mac computer. Then there is another approach, which was suggested by John Mackenzie (North Carolina State University) at one of the courses he gave, called a "gated Integrator", - I'm not sure of the company who sells these, but I plan to contact him to find out.
*************
I have seen your request for digital image capture (with annotation, file storage, printer etc.). We have just such a system that does all that and some more. We have developed it ourselves and it runs our Hitachi 2300. It has the features you list plus automatic contrast setting, image averaging and is also used with our EDX to obtain element maps. It also captures images from several of our other instruments (Auger and SIMS systems). If you are interested I could send you a demo either by email or on a floppy.
**************
We purchased an Hitachi S3200N variable pressure SEM early this year and have the Quartz PCI system that they sell. It runs on a PC and since I had been a Mac person up until this I had to learn a few new tricks. With windows 95 it was not difficult to get going after a short time. This system only captures images at the photo scan rate so you sit for the same length of time it takes to expose a Polaroid. You can do both at the same time. One thing we don't have yet is the option to "play back" a stored image to the microscopes polaroid camera. We will get this soon. Once the image is captured it is composed of two files, the image file in the TIF format, or you can chose from a list of others, and a text file containing any text associated with the image. It lets you put labels and arrows and do some measurments. You can also rotate things and crop images. It does probably most things one needs. It works like some of the drawing or graphing programs such as Cricket graph or MacDraw. Any labels or text can be permenantly "burned into" the image but can not be changed once this is done. This is somewhat important since when you export the image file the text file must also go along unless it is "burned in". We do work for a variety of users and they may not always like where or the type of label applied. This is a minor logistical problem but a pain in the butt with some of our more "difficult" customers. One odd thing is that the information bar, with micron marker, at the bottom of the image and any labels or text added to the image on the microscope (the 3200 lets you label and measure on its screen, I don't know if your SEM does this) DO NOT capture very well. The characters are incomplete. This has something to do with the way the SEMs character generator produced the letters and figures. They show up fine on polaroids because during the film exposure all characters in the image field and info bar are exposed at the end of the scan. I think this scan is repeated several times to provide complete characters. The PCI system stops capturing once the scan reaches the bottom of the screen therefore the characters only get one pass and show up with some missing spots. I explained and showed this to our Hitachi service engineer and she had never noticed this before and was going to bring it to the attention of the PCI people. The characters that PCI generates are very nice and you can change the font and its size as well as bold or italics. It can store images as a database, which was too much for me, or like regular DOS files. I create a folder for each client and folders inside for various projects or samples from that client. A searchable database is nice but who has time time figure all that stuff out. I have not evaluated any other systems and can't comment on them. Over all the PCI system has met our needs but the problem described above is annoying.
**************
Saw your question on the microscopy listserver and since we have an Hitachi SEM (S-450LB) with a Quartz PCI frame grabber I thought I would respond with some comments. The system works very well for image capture and has considerable versatility. If you have a good service person for installation it can be set up in an afternoon. The biggest problem is getting the right computer box as the two boards are very large. We have a Certified Data pentium computer with four ISA slots and 3 PCI slots, and the boards just barely fit. Things like power supplies and fans can get in the way. My recommendation is to have Hitachi select the computer for you if you are going with this system. The data base associated with the frame grabber is quite powerful and allows a lot of manipulation of images for archiving and processing. It is not the most user-friendly system and I am still trying to figure it out. I have many fourth year undergraduate students using it this semester and teaching them how to run it has been rather frustrating. Once we develop a consistent methodology for storing images from a variety of student users, and write up our own manual things should go easier. The manual that comes with it is written for someone with a good working knowledge of computers, and should be scaled down for people like myself. I looked around for awhile before buying this unit and feel that it is probably the best passive capture system on the market, although the Orion system looks interesting as well and may be cheaper. As far as I can tell both systems do much the same thing, i.e. capture images at variable resolutions that are easily manipulated, and allow playback to the photo recording camera (a useful feature). I'm not sure whether the Orion system comes with a built in data base or whether you have to purchase a separate one. I hope this helps you and don't hesitate to write back with more questions if you have them.
**************
I used thdHitachi PCI system with my S-2500 and loved it. I don't think there is any other system that works as well.
**************
I have experience with the Hitachi system, which is actually made by Quartz Imaging and sold by Hitachi. It can be fitted after-market to any SEM of any make. I have two: one on a Hitachi S-570 (1985) and one on a Hitachi S-2300 (1990). The program, called PCI, runs under Windows 95 and takes any slow scan image into the computer where you can do anything with it you wish. I use it on 17-inch monitor, where it shows the image to a whole class like a 11" X 14" blowup. I print to a laser printer and have cut my Polaroid expenses to about one-third. Everyone likes the ability to put images into documents with "Edit-Copy", "Edit-Paste". There are other systems sold, I have seen them but not used them. GW has one, there is one from Australia and there is one from Europe. I'm sure others will tell you about them. I know it is hard to realize now, but after you have one you will wonder how you managed without.
**************
We sell the Orion system in this country. It, like the Hitach system, is a passive capture system. i.e. it sync's to the line and frame of the microscope and digitized the ouput signal. We have customers who have tried both systems and think that the Orion gives better (less pixel jitter, better S/N) images). The system can capture any size up to 8kx8k. It averages and integrates images to reduce noise. A complete system with 1200dpi printer on a 200MHz Pentium wiht 32 MG RAM sells for about $20,000. We can send you some sample images and lit if you are interested.
**************
DVC Company offers 10 bit real time analog and digital output CCD cameras with standard C mount for coupling, and many different frame grabber boards and software as a system.
**************
Bob - you can get information about the Orion system off of the web at http:\\www.microscop-uk.org.uk,80/prodir/orion1.html
**************
I had a demostration of PCI image system from Hitachi at my E.M. Unit a year ago. I was quiet impressed about how easy to capture the image from my Hitachi S-2500 SEM and replay back to camera for photo quality picture. Now acturally I just purchased the sytem a week ago. I played around a bit and found it is even better than what I thought. In the version 4 of the system , it is more easy to get 3-D image and color the images capatured from SEM. I plan to introduce it to my clients to use it as daily research and get instant images on printer or store in a zip drive 100 MB disk in the new year.
**************
E. Fjeld also is a distributor of the SEMICAPS (PC) imaging system. Another board, DIGISEM, is also available fron the ELMDAS Co. in Alexandria PA. Contact John Best at:
A third option is the 4PI system. They now offer both MAC and PC. Contact Mike Czysz at:
4pi Analysis, Inc. 3500 Westgate Drive, Suite 403 Durham, North Carolina 27707-2534 (919) 489-1757 http://www.4pi.com czysz-at-4pi.com (I think) **************
} I would like to thank Don for posting replies to his query about } sensitivity to acrylics. No, Don, it shouldn't be "Whew! That's } over...!" (I know what you mean), for anyone in the microscopy field } who is exposed to chemicals, radiation, and/or any other hazard - it } should be just the beginning!. I would ask everyone to consider that } we do no know what the long term effects of exposure to many } individual chemicals or combinations of chemicals can have on your } health. Are you willing to risk a terminal illness when it is } relatively easy for you can work in a fume hood and wear personal } protective gear? I would hope not! } } Damian Neuberger
Has anyone done an epidemiological survey on life expectencies and causes of death (besides frustration) of microscopists? Life vs Materials? Phil
&&& Illigitimi non carborundum &&&&&&&& Philip Oshel Microscopy Station A PO Box 5037 Champaign, IL 61825-5037 (217)244-3145 days (217)355-3145 evenings oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
CAN SOME ONE TELL ME , DOES A 530MHz PUPPY HAVE A BYTE ?
HAPPY NEW YEAR
Patrick Echlin Multi-Imaging Centre CambridgeOn Tue, 7 Jan 1997, Nathan O'Connor wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } X704, which would be a 530 MHz puppy to be fitted into the next } What would the cost of a system with that kind of chip, the data bus of an SGI, } and the rest of the accessories be like? } } Nathan }
} Are those Pentium Pro chips? That's what they were looking at in the article I } read.
Nathan,
The motherboard that I purchased supports Pentium Pro chips, but out of economics, and at that time a scarcity in that chip, I elected to go the cheaper route. I'll eventually upgrade to the superfast chips when the price goes down, but for the time being, this system is more than adequate for my needs.
-=W.L. Steffens=- Department of Veterinary Pathology College of Veterinary Medicine University of Georgia http://www.vet.uga.edu/wls/steffens.html
Can anyone tell me a good way to quantify Pb concentration in a "galvanized" coating on steel. My understanding is that Pb is distributed in the coating such that focused micro beam techniques may have difficulty sampling enough territory to be accurate. We have SEM/EDS, AES, EPMA, XRF and ICP here in the lab.
Any "tricks" or experience on this matter would be greatly appreciated.
thanks Terry R. McCue Babcock & Wilcox Research 1562 Beeson St. Alliance, Ohio 44601 Phone: 330-829-7427 Internet: terry.r.mccue-at-mcdermott.com Fax: 330-829-7831
Tina, I saw Mary Mager's comments about rapid (TV) scan giving a poor image from an SEM. She is right, because the signal-to-noise is much worse at TV rates. Now, I have a question. Actually, three questions. (1) Would it be possible to collect several successive images at TV rates and average them to improve the signal-to-noise, and thereby obtain a decent image? (2) Can this be done with a SNAPPY frame-grabber? (3) Is there software which will do this? Dr. Dennis B. Barr | 333 DDDDD 333 DDDDD Eastman Chemical Company | 3 3 D D 3 3 D D Microscopy and Morphology | 3 D D 3 D D Research Laboratory | 33 D D 33 D D P.O. Box 1972 | 3 D D 3 D D Kingsport, TN 37662-5150 | 3 3 D D 3 3 D D | 333 DDDDD 333 DDDDD Voice: 423/229-2188 | E-mail: dennbarr-at-eastman.com | IMAGE IMAGE FAX: 423/229-4558 |----------------------------------------------------- (Cross your eyes to see the 3 dimensional image above)
} Now, I have a question. Actually, three questions. (1) Would it be } possible to collect several successive images at TV rates and average them } to improve the signal-to-noise, and thereby obtain a decent image?
Yes, we do this with our video-rate intensified CCD. We can also do a background subtraction, running average, exponential fall-off average and other processing.
} (2) Can } this be done with a SNAPPY frame-grabber?
I don't know. Most, if not all, our capabilities are built into the Perceptics PixelTools 425 Series video capture board.
} (3) Is there software which will } do this?
NIH Image and others can do the processing. The real-time display of the processed images at video rates, however, is more difficult. The Perceptics board does this in our application. I don't know whether the combination of hardware you want and some processing software can do this. Yours, Bill Tivol
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Reply to: RE} } Sgi and Pentium Pro, also MAC??!!
} Date: 1/7/97 1:48 PM } From: jan ringnalda
} There was also a report about a no-nonsense new Motorola chip, the } X704, which would be a 530 MHz puppy to be fitted into the next } generation MACS. Maybe we can go back to user-friendly systems with } excellent performance... Windows just doesn't quite cut it yet.
} Cheers, Jan
Thanks Jan,
I was about to spend my money on a mere 500MHz DEC Alpha running Windows NTwith 512MB RAM, 4.3GB fast-wide SCSI disk and 21" 0.25mm 1600x1200 monitor . . .
One of my zip disk which contains important data was unreadable. The zip drive is working all right. I tried to call the Iomega com. but they are hard to reach. Is anyone has experience on this type of problem? Is any way I can recover some of my data files? Thank you.
**************************** * Maoxu Qian, Ph.D. * * Dept of MSE, box 352120 * * University of Washington * * mxq-at-u.washington.edu * * (206)543-1514(phone) * * (206)543-3100(fax) * ****************************
I just checked prices of the dual pentium pro systems available from Micron Electronics. Systems with 2 200Mhz processors can be had from about $3500 and their most expensive configuration is about $7500. The upgrade to the second processor from a single processor system is $700. Obviously there are graphics workstations available with more capabilities and higher prices (probably including the system compared in BYTE). I checked the prices from Micron because I own a couple of their computers. If you want more info on similar systems I suggest you take a look at Intel's web site. They have links to a number of companies that sell such systems. Mike
Mike Bench Characterization Facility Center for Interfacial Engineering University of Minnesota Voice: (612) 624-6590 Fax: (612) 626-7530 e-mail: bench-at-cems.umn.edu
I just checked prices of the dual pentium pro systems available from Micron Electronics. Systems with 2 200Mhz processors can be had from about $3500 and their most expensive configuration is about $7500. The upgrade to the second processor from a single processor system is $700. Obviously there are graphics workstations available with more capabilities and higher prices (probably including the system compared in BYTE). I checked the prices from Micron because I own a couple of their computers. If you want more info on similar systems I suggest you take a look at Intel's web site. They have links to a number of companies that sell such systems. Mike
Mike Bench Characterization Facility Center for Interfacial Engineering University of Minnesota Voice: (612) 624-6590 Fax: (612) 626-7530 e-mail: bench-at-cems.umn.edu
I embedded tissue culture cells on glass coverslips in EMBed (Epon-like) and now cannot remove the glass with thermal or mechanical shock.Please let me know if you have a trick for separating glass from epoxy without destroying the epoxy. I also have experiments running with plastic substrates but would like to recover this one sample. Note: the coverslips were coated only with poly-D-lysine and embedded by inverting a Beem capsule filled with epoxy on the coverslip. Thanks! Larry D. Ackerman (415) 476-8751 Howard Hughes Medical Institute FAX (415) 476-5774 UCSF, Box 0724, Rm U426 533 Parnassus Ave. mishot-at-itsa.ucsf.edu San Francisco, CA 94143
------------ Forwarded Message begins here ------------
Hi, there,
Several people asked me more information on the problem. Sorry I didn't say clearly. This is on a MacIIci mation, When insert the zip disk, it keeps rotating then a click sound and rotating and click sound ...for long time and in the end, the message cames out as "unreadable disk". I tried use Norton utility to recover it but Norton did not recognize zip drive. Is there any utility software can do it? or the disk is physical damaged? The disk was sitting inside of the drive all the time but due to reboot, I have to reinsert it and the problem occurred. Any suggestion? Thank you again.
**************************** * Maoxu Qian, Ph.D. * * Dept of MSE, box 352120 * * University of Washington * * mxq-at-u.washington.edu * * (206)543-1514(phone) * * (206)543-3100(fax) * ****************************
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I embedded tissue culture cells on glass coverslips in EMBed (Epon-like) and } now cannot remove the glass with thermal or mechanical shock.Please let me } know if you have a trick for separating glass from epoxy without destroying } the epoxy. I also have experiments running with plastic substrates but would } like to recover this one sample. Note: the coverslips were coated only with } poly-D-lysine and embedded by inverting a Beem capsule filled with epoxy on } the coverslip. Thanks! } Larry D. Ackerman (415) 476-8751 } Howard Hughes Medical Institute FAX (415) 476-5774 } UCSF, Box 0724, Rm U426 } 533 Parnassus Ave. mishot-at-itsa.ucsf.edu } San Francisco, CA 94143 } } Larry --
Our method of last resort is to jam the tip of a dissecting needle under an exposed edge of the coverslip, then pry up gently trying to lift the the coverslip off of a near-by region. You end up damaging some of the specimen this way, but with some luck and careful trimming at the microtome one can still get some nice areas to section.
Hope this helps, Greg Martin Dept. of Cell Biology and Anatomy Johns Hopkins School of Medicine
At 10:16 AM 1/8/97 -0500, you wrote: } Can anyone tell me a good way to quantify Pb concentration in a } "galvanized" coating on steel. My understanding is that Pb is } distributed in the coating such that focused micro beam techniques may } have difficulty sampling enough territory to be accurate. We have } SEM/EDS, AES, EPMA, XRF and ICP here in the lab. Dear Terry, My recommendation would be EPMA with a de-focused beam. Spread the beam to 2 to 5 microns in diameter and sample at least 10 random spots. Be sure to test the substrate element (i.e. Fe) to track differences in coating thickness, which will affect apparent Pb concentration. A known standard would also be very helpful. Good luck, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
} Now, I have a question. Actually, three questions. (1) Would it be } possible to collect several successive images at TV rates and average them } to improve the signal-to-noise, and thereby obtain a decent image? (2) Can } this be done with a SNAPPY frame-grabber? (3) Is there software which will } do this? Dear Dennis, Yes, some frame-grabbers can do frame averaging on several successive frames (I have seen up to six). This is a hardware characteristic of the more expensive frame grabbers. The TV rate is too fast for software. I don't think the Snappy has this characteristic. The image must be stable, which TV rate images on SEM sometimes aren't,a nd of course the resolution will be limited to the 525 lines of NTSC. I think the Snappy would be a good Light Microscope solution, but less so for SEM. Good luck, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
At 12:59 PM 1/8/97 -0800, Larry Ackerman wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Larry, Have you tried cooling the glass coverslips on the surface of a piece of dry ice? It (the coverslip) usually pops off leaving the embedded cells behind. Rosemary
Seasons greetings to TEMists, SEMists & probers. We at the University of Durban-Westville are planning to purchase a new TEM in the near future. YThe machines under consideration at the moment are the Philips 208S or if finances permit Jeol 1010.
To assist with our selection, I would appreciate responses from any of you with experiences (good or bad) with either instrument. Particular areas of interest are: 1. Quality control and reliability of electronic and mechanical components. 2. Quality of images at all maginifications and instrument stability.
We are replacing a still functional Philips 301 - any offers!
To avoid cluttering up the list, please contact me at my e-mail address.
Best wishes for 1997
Mike Gregory EM Unit University of Durban-Westville Durban South Africa
I would appreciate suggestions for examining 80 micron eggs in seawater or distilled water by SEM where the user wants to compare fresh egg surfaces (therefore cryo?) with conventionally fixed, dehydrated and critically point dried eggs.
The idea was to use cryoSEM but there are problems. I have tried freezing (in ethane) on a thin smear of Tissue Tek OCT - droplet on stub and then wiped off with cover slip (not easy in itself?), but the eggs sank, never to be seen again!
I have also frozen them just mounted in a water film, but some have simply sheared off the standard smooth stub - should I try poly-L-lysine or super glue?
A separate problem seems to be that while the crustacean in question is marine/estuarine and the eggs stand fresh water for a while (therefore for rinsing), even after 3 or 4 distilled water changes I still see a fine, dried layer of salt over everything. The rinsing is done in a watchglass and the water was almost completely removed with an autopipette before refilling.
One idea not tried yet is to place an egg on a millipore filter, drain on filter paper, place on stub in cryoSEM airlock cold-stage and freeze in situ. But with or without vacuum?
It seems a crazy world when it is easier to ask internet friends than it is to search through a bucket a mud and then do the experiments on the method to use!
Any comments would be appreciated. Take up gardening?
Thanks in advance - Keith Ryan
++++++++++++++++++++++++++++++++++++++++++++++++++ Plymouth Marine Laboratory, Citadel Hill, Plymouth, Devon PL1 2PB, England
e-mail: k.ryan-at-pml.ac.uk PML web site: http://www.npm.ac.uk/pml ++++++++++++++++++++++++++++++++++++++++++++++++++
One method we use to remove the glass coverslip is to place the block with the attached coverslip into hydrofluoric acid. The acid will slowly dissolve the glass in about an hour. The acid does not dissolve epoxy, and we have not noticed with the embedded cells.
Regards,
John J. Turek, Ph.D. Purdue University Dept. of Basic Medical Sciences Director, Core Laboratory for Image Analysis and Multidimensional Applications (CRISTAL) phone: 317-494-5854 fax: 317-494-0781 email: jjt-at-vet.purdue.edu web: http://vet.purdue.edu/cristal
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello to all~ I have recently joined this list, and found all of the discussion very informative. Since many questions discussed here deal with finding suppliers, I thought I would let you know of ASM International's online version of our "testing buyer's guide". It can be found at http://www.asm-intl.org/testing
The site gives contact details for over 400 companies providing services related to testing, including microscopy.It is based on information compiled by the editors of Advanced Materials & Processes, ASM's monthly magazine.
Best wishes for 1997~
Leslie H. Chom LHChom-at-po.asm-intl.org Information that Goes to Work-from the Materials Information Society ASM International http://www.asm-intl.org (ph) 216-338-5151 Ext 510 (fx) 216-338-4634
Greetings, and best wishes for the New Year. Volume 2, Number 1, of the PENSEES NOUVELLES is now on line at
http://www.emory.edu/PATHOLOGY/PENSEES
Contents include some medical doggerel, two book reviews (one about histologic techniques with microwaving, and one on golf), and the continuation of the review of the Robert Trent Jones Golf Trail courses.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
To look at surfaces of small samples, roughening the stub surface with fine sandpaper will help keep the ice on the stub during cryo, but we went into overkill and machined a slight "well" in the surface of the stub and then roughed it up. To get a cross sectional view, we drill several small wells (1/16 in.dia x 1/8 in. deep) in the stub. Overfill the wells (this can be tricky) with a high concentraion of sample in water leaving a slight droplet (protruding meniscus) of sample protruding above the well. Sometimes a little vaseline on the surface will help keep the droplets from running together. Freeze and shear off the droplets.
For the dry eggs, we would use about a dozen (pelco #16079) adhesive tabs stuck on the surface of a clean glass slide. Dissolve these into 20 ml chloroform. Place a drop or two of this solution on the surface of the stub (or coverslip for a smoother background surface), spread evenly and drain the excess with the corner of a kimwipe. This usually produces a very thin layer of adhesive strong enough for samples up to a few hundred microns.
As for the salt on the surface after 3 or 4 washes, not much help I'm afraid, just a thought. Is there a mucosal surface layer that the distilled water is drawing the salt out of ?? If so, they might need an EDTA treatment first to remove mucus. Then again, this mucus might be the surface you need to see??? This might be a case of "a rock and a hard place", but sucessful cryo should tell you what is on the surface if you can get a fracture through an egg.
Hope this is helpful.
Cheers :) George Department of Earth and Atmospheric Sciences University of Alberta Edmonton, Alberta T6G 2E3 Canada ph: 403-492-5746 fax: 403-492-2030
} Hi, there, } } Several people asked me more information on the problem. Sorry I } didn't say clearly. This is on a MacIIci mation, When insert the zip disk, } it keeps rotating then a click sound and rotating and click sound ...for } long time and in the end, the message cames out as "unreadable disk". I } tried use Norton utility to recover it but Norton did not recognize zip } drive. Is there any utility software can do it? or the disk is physical } damaged? The disk was sitting inside of the drive all the time but due to } reboot, I have to reinsert it and the problem occurred. Any suggestion? } Thank you again. }
Maoxu,
Rebooting after computer crash can damage the important directory/b-tree area on disk. I do have experience of this kind of damage. The damage didn't recover by Norton Utility. Now I always take the disk out before rebooting. After rebooting, insert and eject it, then insert it again. Now it works normally.
Ya Chen
Ya Chen
*** My email address has been changed to: ychen14-at-facstaff.wisc.edu *** ========================================================================= \ / Integrated Microscopy Resource (IMR)--
\ / __ an NIH Biomedical Research Resource TEL : 608-263-8481 \/ / / University of Wisconsin-Madison FAX : 608-265-4076 / / / 1675 Observatory Drive #159 Email1:ychen14-at-facstaff.wisc.edu / /__/_ Madison, WI 53706 Email2:chen-at-calshp.cals.wisc.edu ========================================================================= IMR WWW Home Page: http://www.bocklabs.wisc.edu/imr.html
I agree wholeheartedly with Mary Mager on the suitability of Snappy for SEM. I'd only recommend them in two cases, as I've discussed with some of you privately.
1. Where there is absolutely no money for a better solution AND the user has lots of time to attempt offline averaging solutions AND the samples aren't very demanding.
2. Where the SEM has built in frame averaging capability AND the only application for the instrument is simple photographic documentation.
I think Tina fits into catagory one. I'm basing this partially on the images at her website. If a large percentage of her work is at very low magnification, stability becomes less of an issue and she has the possibility to average images off-line.
Dear Fellow Microscopists: We are considering of installing an Automatic Liquid Nitrogen Filling System for our SEM. The only source we came across is VBS Industries, Inc. I wonder if any of you would like to share with me your experience with any automatic LN2 filling system. Are they dependable? Are there other sources that sell similar type system? Any feed back will be greatly appreciated.
Pearl W. Yip Rome Lab yip-at-maxwell.rl.plh.af.mil
There was a long thread on this subject in the early days of the listserver. Perhaps it resides in Nestor's or the Florida archives??
I put in my comment that I would never have auto LN fillers again. We had them on TEMs and SEMs in the 70s and early 80s and they all failed catastrophically. (I guess there is no simple, benign failure mode when the possibility of emptying a large LN cow over an instrument and into an unoccupied room exists)!
Long term reliability under extreme temperature excursions seemed to be the problem.
However, to be fair, we are talking 20+ year old technology. This is the wonderful 90s...
One way to examine your marine eggs would be to use the high-pressure freezer to freeze a suspension of washed eggs in distilled water (a roughly 200 micrometer thick plug, 1000 or more micrometers in diameter). Then fracture, etch, and coat as for a TEM replica but use the cryoSEM to look at the rough surface of the fracture. By regulating the amount of etch you could see the surface of the egg as well as cross-sections of the layers of the walls. Paul Walther's articles on double layer coating may be helpful.
Jacob
Jacob Bastacky, M.D. Room 116 Donner Laboratory Lawrence Berkeley Laboratory EMail: sjbastacky-at-lbl.gov University of California Phone: (510) 486-4606 Berkeley, California 94720 Fax: (510) 486-4750
Dear Larry: We've had consistent success removing cover slips from Epon capsule-embedded monolayers with a combination of the methods mentioned by Gib and Greg. We dip the epon block with attached glass or plastic fragments into liquid nitrogen (block can already be attached to chuck). Then using a razor blade gently pry up the rapidly-warming fragment. Sometimes it takes a couple of cooling/warming cycles to obtain sufficient area. Don Gantz Boston Univ Med School gantz-at-med-biophd.bu.edu
Please post the attached ad for a Research Electron Microscopist as soon as possible. If you have any questions, please call me at (217) 244-2944, fax (217) 244-2946.
University of Illinois at Urbana-Champaign Materials Research Laboratory
RESEARCH ELECTRON MICROSCOPIST
The Materials Research Laboratory at the University of Illinois is seeking an experienced electron microscopist as a staff member in the Center for Microanalysis of Materials. The Center is a major research facility with eight electron microscopes as well as instruments in surface microanalysis, x-ray diffraction and other analytical techniques.
The person will work mainly on STEM and TEM but should have experience and the flexibility to work on other techniques if needed. The Center has six TEMs. We are particularly interested in hiring a person with experience in working with field emission TEMs. The Center currently has a VG HB 501 and is expected to purchase a new FEG-TEM soon. The person appointed will have primary responsibility for these instruments. Experience in EDX or EELS is also important. The main responsibility of the position would be to facilitate the research of approximately 100 users yearly on TEM and STEM. There will also be ample opportunity to carry out interactive research in the facility with the wide range of research programs.
This position requires a Ph.D. with at least three years experience with electron microscopes. Salary is commensurate with experience and qualifications.
This is a regular full-time appointment with standard university benefits. The person appointed should be able to begin work between June and August, 1997. In order to ensure full consideration, applications must be received by March 15, 1997. Please send letter of application, resume, and names and addresses of three references to J. A. Eades, c/o Donna Jacobs, University of Illinois, Materials Research Laboratory, 104 South Goodwin Avenue, Urbana, Illinois 61801, phone (217) 244-2944. For technical information, call J. A. Eades at (217) 333-8396.
The University of Illinois is an Affirmative Action/Equal Opportunity Employer.
Donna Jacobs MRL Administration University of Illinois 104 South Goodwin Avenue Urbana, Illinois 61801 Phone (217) 244-2944 Fax (217) 244-2946 email - jacobs-at-uimrl7.mrl.uiuc.edu
} } Date: Tue, 7 Jan 1997 00:41:06 -0500 } } From: PHOBOS11-at-aol.com } } Subject: Wanted Balzers Electronics } } To: Microscopy-at-Sparc5.Microscopy.Com } } Status: RO } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi Al,
We do have a new Balzers EVM-052 E-beam gun power supply. Would you please give us an offer.
Have a nice day.
Ya Chen
Ya Chen
*** My email address has been changed to: ychen14-at-facstaff.wisc.edu *** ========================================================================== Cryo/SEM Coordinator Integrated Microscopy Resource (IMR)-- III M M RRRRRR an NIH Biomedical Research Resource I M M M M R R University of Wisconsin, Madison, WI I M M M RRRRRR 1675 Observatory Drive #159 I M M R R Madison, WI 53706, USA I M M R R TEL : 608-263-8481 I M M R R FAX : 608-265-4076 III M M R R Email:ychen14-at-facstaff.wisc.edu ========================================================================== IMR WWW Home Page: http://www.bocklabs.wisc.edu/imr.html
} Dear All } I would appreciate suggestions for examining 80 micron eggs in seawater or distilled water by SEM where the user wants to compare fresh egg surfaces (therefore cryo?) with conventionally fixed, dehydrated and critically point dried eggs. } .....
} I have also frozen them just mounted in a water film, but } some have simply sheared off the standard smooth stub - } should I try poly-L-lysine or super glue? } ........
} A separate problem seems to be that while the crustacean } in question is marine/estuarine and the eggs stand fresh } water for a while (therefore for rinsing), even after 3 or 4 } distilled water changes I still see a fine, dried layer of } salt over everything. The rinsing is done in a watchglass } and the water was almost completely removed with an } autopipette before refilling. } } } Thanks in advance - Keith Ryan } } ++++++++++++++++++++++++++++++++++++++++++++++++++ } Plymouth Marine Laboratory, Citadel Hill, } Plymouth, Devon PL1 2PB, England } } e-mail: k.ryan-at-pml.ac.uk } PML web site: http://www.npm.ac.uk/pml } ++++++++++++++++++++++++++++++++++++++++++++++++++
Keith,
Here is my 2 cents.
1. Coating of poly-L-lysine do help the sample adhere to substrate.
2. Did you have tried freeze-substitution followed by either fast-freezing again, freeze-drying, cryo-coating, and cryo-SEM observation, or dehydration, critical point drying, and SEM observation at RT?
3. I agree with George Braybrook's comment that there is a mucous layer on the egg surface.
With best wishes!
Ya Chen
Ya Chen
*** My email address has been changed to: ychen14-at-facstaff.wisc.edu *** ========================================================================= \ / Integrated Microscopy Resource (IMR)-- \ / __ an NIH Biomedical Research Resource TEL : 608-263-8481 \/ / / University of Wisconsin-Madison FAX : 608-265-4076 / / / 1675 Observatory Drive #159 Email1:ychen14-at-facstaff.wisc.edu / /__/_ Madison, WI 53706 Email2:chen-at-calshp.cals.wisc.edu ========================================================================= IMR WWW Home Page: http://www.bocklabs.wisc.edu/imr.html
It occurred to me that to be sure that your treatments did not introduce any artefacts on the egg surface you might try a Wild (Leica) Elsam accoustic microscope. I'm not very familiar with them but there is the potential (at least) to look at them in seawater or other isotonic solution.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear All
I would appreciate suggestions for examining 80 micron eggs in seawater or distilled water by SEM where the user wants to compare fresh egg surfaces (therefore cryo?) with conventionally fixed, dehydrated and critically point dried eggs.
I was interested to see your reference to removing mucus with EDTA. Would you be so kind to offer a little more info on this. We are starting to do more work on cilial tracts on the head tentacles of micromolluscs for one of our malacologists and often have problems with mucus.
Cheers,
Geoff Avern Microscopy Laboratories Australian Museum Sydney, Australia
As for the salt on the surface after 3 or 4 washes, not much help I'm afraid, just a thought. Is there a mucosal surface layer that the distilled water is drawing the salt out of ?? If so, they might need an EDTA treatment first to remove mucus. Then again, this mucus might be the surface you need to see??? This might be a case of "a rock and a hard place", but sucessful cryo should tell you what is on the surface if you can get a fracture through an egg.
Hope this is helpful.
Cheers :) George Department of Earth and Atmospheric Sciences University of Alberta Edmonton, Alberta T6G 2E3 Canada ph: 403-492-5746 fax: 403-492-2030
} We are considering of installing an Automatic Liquid Nitrogen } Filling System for our SEM. The only source we came across is VBS } Industries, Inc. I wonder if any of you would like to share with me your } experience with any automatic LN2 filling system. Are they dependable? Are } there other sources that sell similar type system? Any feed back will be } greatly appreciated. } Dear Pearl, Our automatic LN2 controllers are from Torr Vacuum, Inc. We have had several disasters: 1) The LN2 source--a very large tank about 200 m from our lab--would occasionally run dry; then the solenoid would over- heat. Usually it would open, but occasionally it would fuse and draw current until someone noticed (see #3). 2) The filler shut-off failed on the system attached to our EDS detector. LN2 poured into and over the dewar until the outer bottle deformed and ice formed all over the in- side and outside of the system. We had to warm up everything, dry every- thing out and re-form the outer bottle--it had a concave bottom originally, but was convex after the disaster. Luckily, everything worked out well, and the detector is still functioning a decade later with its specified resolution. 3) For some reason--not an empty LN2 tank--the solenoid on the line on the 200 kV TEM fused and heated the plastic foam insulation starting a fire. The fire was, fortunately self-limiting; the event occurred after hours on a Friday before a holiday weekend and was not discovered until the next Tuesday. We have not lost any expensive equipment, but there was certainly the potential for such losses. I don't think our problems were the fault of Torr Vacuum; they seem to be inherent in automatic LN2 systems. If you can avoid such systems, I would reccommend you do so. Good luck--especi- ally if you must use them. Yours, Bill Tivol
} Is there a best way to clean the specimen holder on the TEM? How do most people } clean their holders? } Dear Linda, We put our stage tips in a plasma cleaner for ~20 min. That seems to get the petrified grease off, and the method can be used on all tips, not just those constructed of a single piece of metal. It works on our tilt-rotation tip (mostly aluminum), our double-tilt tip (mostly stainless steel) and our aperture holders (phosphor bronze). Yours, Bill Tivol
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } TEM specimen holder cleaning 1/9/97 12:24 PM } } Is there a best way to clean the specimen holder on the TEM? How do most people } clean their holders? } } I normally use Q tip with a little bit of Wenol to clean the holder first. Then use Kimwipes to rub over entire surfce. Sonicate the holder in the acetone bath for 10 min and once again in the fresh acetone bath for another 10 min. After that use air gun to dry it. That is all I do.
*********************************************** * Ming H. Chen, PhD * * Medicine/Dentistry Electron Microscopy Unit * * University Of Alberta. * * Edmonton, Alberta, Canada * ***********************************************
My recommendation would be to use a mixture of Tissue-Tek and carbon dust (from carbon rod sharpener) paste on your cryo-stub. In this mixture the eggs will not sink fast, therefore will have time to carry out the freezing process. I have great LTSEM results using this mixture with unfixed single cell culture and bacteria.
The separate problem about the fine layer of salt, (if the osmatic change is all right!) use larger amount of distilled water to rinse for longer time.
Good luck,
Laszlo J. Veto Electron Microscopy and Image Analysis Pacific Agri-Food Research Centre AAFC vetol-at-em.agr.ca
My recommendation would be to use a mixture of Tissue-Tek and carbon dust (from carbon rod sharpener) paste on your cryo-stub. In this mixture the eggs will not sink fast, therefore will have time to carry out the freezing process. I have great LTSEM results using this mixture with unfixed single cell culture and bacteria.
The separate problem about the fine layer of salt, (if the osmatic change is all right!) use larger amount of distilled water to rinse for longer time.
Good luck,
Laszlo J. Veto Electron Microscopy and Image Analysis Pacific Agri-Food Research Centre AAFC vetol-at-em.agr.ca
On Thu, 9 Jan 1997, Pearl Yip wrote: } Filling System for our SEM. The only source we came across is VBS } Industries, Inc. I wonder if any of you would like to share with me your } experience with any automatic LN2 filling system. Are they dependable? Are } there other sources that sell similar type system? Any feed back will be } greatly appreciated.
I agree with Ron and Bill, they are not dependable. Anything that has moving parts or electronics is not dependable. If you must keep something cold all the time (much better than temp cycling, IMHO), I recommend that you design and build a large LN dewar that keeps the thing cold that you periodically refill every few days or so - much like a dewar for an EDS detector. There are companies out there that will work with you on this if you don't have the facilities to build your own.
Excellent cryo-fracture images of single cells can also be produced, using the Tissue-Tek/Carbon mixture. (My earlier note) It is also a good adhesive, especially on the rough cryo-stub surface. George had very good ideas on cryo-stub surface preparation. This mixture also fractures well, exposing ALL inner surface of the fractured eggs "embedded" in it. I hope it will work well for you.
Regard,
Laszlo
Laszlo J. Veto Electron Microscopy & Image Analysis Pacific Agri-Food Research Centre, AAFC Ph: 250-494-7711 e-Mail: vetol-at-em.agr.ca
Linda Iadarola requested information on cleaning TEM specimen holders. A commonly used technique is to polish with a metal polish such as Wenol or Pol then either wipe or rinse in methanol. Extreme care does need to be taken to avoid trapping the polishing paste in the crevices of the holder. Also, depending on the type of specimen holder, ultrasonic cleaning must NOT be used since the potential exists to weaken or break epoxy bonds (particularly in the case of cyro holders).
Another possibility is to plasma clean the holder. Most of the contamination resident on holders is organic (hydrocarbon). An air or oxygen/argon plasma is quite effective in reducing this contamination. The plasma creates disassociated oxygen which chemically combines with the carbonaceous material and reduces it to CO, CO2 and H2O. Depending on the ion energies used, cleaning can occur without adversely effecting the specimen holder.
Should you have any further questions, please do not hesitate to e-mail me directly.
Best regards,
Paul E. Fischione
E.A. Fischione Instruments, Inc is the manufacturer of the Model 1400 Plasma Cleaner.
G'day everybody and Happy New year! Would any of you kind electron microscopists have looked at the growth rings in seal teeth or any mammalian teeth, please? And how did you do that? And is there a relationship between the ratio of cementum and dentine to the age of the mammal? Thank you kindly Cheers Gerry nash
Ms Geraldine Nash Electron Microscopist
EM Unit Australian Antarctic Division Channel Highway Kingston Tasmania 7050 Australia Ph: + 61 03 62 323 322 Fax: + 61 03 62 323 351
} I'm in a twisty little maze of references, all different. } } Can anyone tell me the full name of Phil. Mag.? } } Thanks in advance, } } Rick Mott
Philosophical Magazine? Your nearest university library should have a reference work that lists periodical names and official abbreviations. "Should" because there *is* such a thing, and they ought to have it. Phil
&&& Illigitimi non carborundum &&&&&&&& Philip Oshel Microscopy Station A PO Box 5037 Champaign, IL 61825-5037 (217)244-3145 days (217)355-3145 evenings oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
I would appreciate suggestions for examining 80 micron eggs in seawater or distilled water by SEM where the user wants to compare fresh egg surfaces (therefore cryo?) with conventionally fixed, dehydrated and critically point dried eggs. . . . . ************************************* You have two major problems: Sticking and retaining the processed eggs onto a substrate prior to coating and earlier, removing all traces of salt from the eggs . You could try poly-l-lysine. The old technique, which works well for specimens of that size range, is a drop of a sticky solution onto a substrate. After solvent evaporation a very thin "permanently" sticky layer remains. The solution is prepared by dissolving the gum of clear sticky tape in chloroform overnight. Push a couple of meters of tape into a small jar and add about 50 ml of chloroform. If you require a very clean background the solution can be millipore filtered. If some low power images are needed, than freshly cleaved mica gives a very clean background. At zero tilt, the mica is very dark in secondary mode.
Removing salt from marine specimens can be very difficult. Leaving the specimen for a lengthy period in water, even after fixation, may damage structures. An appropriate concentration of ammonium acetate provides the right molarity and the aqueous (or ethanol) solution leaves no residue. Others have referred to mucous which may or may not be removed. EDTA fell out of favour as a decalcifier some time ago. For the same reasons I would prefer a broad spectrum enzyme. I used a snail enzyme many years ago for removing mucous, but your local biochemist should be able to advise. Hope this is some help with this difficult problem. Happy New Year Jim Darley Probing & Structure (Microscopy Supplies & Accessories) PO Box 111, Thuringowa QLD 4817 Australia
Phone +61 77 740 370 Fax: +61 77 892 313 A great microscopy site: http://www.ultra.net.au/~pns/
Please, can you tell me how i can participate to this helpfull Newsgroup ? Thanks Christian (student preparing a Microscopist PhD in Nantes University ) .
} Dear Fellow Microscopists: } We are considering of installing an Automatic Liquid Nitrogen } Filling System for our SEM. The only source we came across is VBS } Industries, Inc. I wonder if any of you would like to share with me your } experience with any automatic LN2 filling system. Are they dependable? Are } there other sources that sell similar type system? Any feed back will be } greatly appreciated. } } Pearl W. Yip } Rome Lab } yip-at-maxwell.rl.plh.af.mil
Pearl,
As others have said, these systems do seem to unreliable and considering the value of the equipment they are attached to, I would not trust them. I would even be wary about LN2 level alarms - they seem to be reliable, but do you really want to have the safety of perhaps 500k dollars or more of equipment depending on one?
Anyway, what is the problem with a regular manual schedule of re-filling?
You don't actually say what you are filling with LN2.
If it is an EDX detector, I would suggest that if you are really concerned, you should have a routine for users to disconnect the HT from the detector. Then if the detector should run dry, there is no risk of damage (although, to be honest, I have, more than once, had an EDX detector go dry while the HT was on and survive apparently unharmed, but I don't recommend anyone trying it). However, this might require that you allow the detector to restabilise following reconnection of the HT.
If you are filling LN2 traps on the vacuum system, then I would caution you about running them continuosly anyway - over a period of time this will actually degrade your vacuum. All cold traps should be allowed to warm to room temperature at least once a week and the vacuum stabilise. If not, you will gradually get a build up of ice and other contaminants on the traps which, eventually will outgas at a sufficient rate to contaminate your system.
} } TEM specimen holder cleaning 1/9/97 12:24 PM } } Is there a best way to clean the specimen holder on the TEM? How do most } people } clean their holders?
Don't :)
No part of a specimen holder that goes into the vacuum of a TEM needs to be, or should be touched by dirty fingers (or anything else). If you follow appropriate handling procedures, in the majority of cases, holders do not need cleaning.
They may often 'look dirty' but this is usually some sort of oxidation and doesn't cause any problems in the TEM. Unless a particular specimen rod is, without question, causing contamination problems when you do microanalysis or microdiffraction, and the problems really are only apparent with that specific rod, then 'if it ain't broke, don't fix it'.
Certainly, there should be no necessity for a regular cleaning routine and in general, I have never cleaned holders for which I have had responsibility. The only exceptions are the external O-ring seal on the barrel and the jewel bearing on the tip.
Having said that, problems do occur. Simple holders (single tilt) can usually be cleaned successfully by ultrasonic in a solvent, rinse in distilled water and warm air blow dry. However, more complex holders may be almost impossible to clean fully - it is difficult to fully penetrate all the crevices and internal spaces effectively and solvent/ultrasonic may weaken or damage expoxy joints and seals. Plasma discharge is pretty effective, but again is unlikely to fully penetrate internal spaces - although if you have serious contamination in an internal space then whoever is responsible probably needs introducing to a few of life's realities - try to find an old HT tank for them to clean!
Whatever the problem, don't use wehnol or similar abrasive metal polishes - if it needs something that powerful to remove the dirt, then it wasn't a problem to start with - but it may be after you have filled the crevices with metal polish.
Minor contamination of holder tips by specimens can sometimes be a problem. Usually, this can be cured by leaving the holder, without a specimen, in the TEM contiuosly for a long period - say a weekend - and it will pump clean.
The only exception to all the above is cryo-holders. They frequently get horribly dirty. Often, however, it is only the tip region that is the problem. If you don't have access to a suitable plasma system, then just the tip can be suspended and ultrasoniced in a solvent - also, check with the manufacturer regarding cleaning. You may find that you have to start by removing the worst with wooden cocktail sticks. You will avoid the worst problems if you can get users to remove specimens from the holders while still frozen.
Regards, Larry Stoter
--------------------------------------------------------------- Dr L. P. Stoter Technical Editor, MICROSCOPY & ANALYSIS Technesis 17, Rocks Park Road email: LPS-at-teknesis.demon.co.uk Uckfield, E. Sussex Phone: +44 (0)1825 766911 TN22 2AT Fax: +44 (0)1825 766911 United Kingdom ---------------------------------------------------------------
} } I'm in a twisty little maze of references, all different. } } } } Can anyone tell me the full name of Phil. Mag.? } } } } Thanks in advance, } } } } Rick Mott } } Philosophical Magazine? } Your nearest university library should have a reference work that } lists periodical names and official abbreviations. "Should" because there } *is* such a thing, and they ought to have it. } Phil } } &&& Illigitimi non carborundum &&&&&&&& } Philip Oshel } Microscopy } Station A } PO Box 5037 } Champaign, IL 61825-5037 } (217)244-3145 days } (217)355-3145 evenings } oshel-at-ux1.cso.uiuc.edu } *** looking for a job again ******************
But beware that there are Phil. Mag. A and Phil. Mag. B. I don't know when each issue was split into two, but is was some time ago.
A postdoctoral position is available immediately in the department of cell biology at the University of Pittsburgh. This position within the Muscular Dystrophy Research Center and Imaging Center will be focussed toward using microscopy to study the dystrophin cytoskeleton in skeletal muscle, the ultimate goal being to optimize therapeutics (gene delivery) currently under development within the center. Initially the project will use live cell, confocal, and EM methods to study the expression, and deposition of components of the dystrophin cytoskeleton (dystrophin, sarcoglycans, dystroglyans and the ECM) and their role in the development and maintenance of muscle fiber structure. This position is funded for 3 years
This position requires a Ph.D.and experience in microscopy (the specific field is unimportant) This is a regular full-time appointment with standard university benefits
The University of Pittsburgh is an equal opportunity employer
Please respond by e-mail to swatkins-at-pitt.edu
-- ------------------------------------------------- Simon C. Watkins Ph.D Associate Professor Director SBIC University of Pittsburgh Pittsburgh PA 15261 tel 412-648-3051 fax 412-648-2004 -----------------------------------------------
-- Leon A. Metlay, M.D.,Associate Professor of Pathology and Laboratory Medicine University of Rochester Medical Center Phone: (716) 275-5691 P.O. Box 626 Fax: (716) 273-1027 Rochester, NY 14642 lmetlay-at-acu.pathology.rochester.edu http://www.urmc.rochester.edu/smd/pathres/URPLM.html "Most ass drivers are evil, most camel drivers are decent, most sailors are saintly, the best among physicians is going to Gehenna, and the best of butchers is a partner of Amalek" -R. Judah, in Mish. Kidd. 4:14
The filler for an LN trap over the DP on our JEOL 2000FX TEM malfunctioned, and a tank of LN2 emptied out all over the diffusion pump. The microscope shut down, luckily valving the column off from the DP. The water lines froze, including the water in the baffle at the top of the pump. This caused the baffle to crack, and when it warmed up again, the pump and reservoir tank flooded with water. The next morning the microscope was found in the OFF condition, and, no problem being evident, was simply restarted. This caused an emulsion of DP oil and water to be sucked into the rough pump, which *was* evident, and the machine was immediately turned off. The damage was done however, and a complete teardown and cleaning was necessary, taking a couple of weeks.
So if you use a filler system, be sure to provide a way to funnel LN away from the microscope, in the event that the solenoid fails to close properly and the tank empties.
Microscopists, at least in the U.S., may want to learn a new acronym. On 7 January I attended a forum at the National Institute for Standards and Technology which started the process to form NACLA, the NAtional Council for Laboratory Accreditation. While it is in its early infancy, this group is seen as a way to resolve the current crisis in laboratory accreditation, which affects more of the members of the microscopy listserver than one might imagine.
Those of you in academic institutions can now lean back, relax and have a good laugh at the expense of the rest of us. The academic world learned long ago that accreditation was a good idea, but it was only going to work if the standards were set high enough and the process was rigorous enough that the results were acceptable to everybody involved. The result is mutual recognition of accreditations done by various bodies, some based on regional associations and others by national groups which derive from various professions. The result is a system that works pretty well.
For three different groups of laboratories, however, accreditations are not mutually recognized, for a variety of reasons. Much of the time of the forum was devoted to describing the problems of testing, analytical and calibration laboratories (medical and environmental laboratories have whole additional layers of problems) operating within manufacturing organizations and doing product-related analytical work, operating as independent laboratories or operating within the Federal government. The crisis is that laboratory folks are spending so much time dealing with duplicate accreditation visits that there is little time left to get any work done. Approximately 150 different organzations in the U.S. accredit laboratories. The current record is a laboratory that holds 102 separate accreditations from various levels and agencies of government, different industrial users of laboratory data and several accreditation associations. Each of these accreditations requires that an audit be conducted, ranging from submission of duplicate documents to be analyzed in the same way to on-site visits taking several days.
Underlying this mess are several types of problems, including the fact that different agencies accredit to different standards, the conflicting requirements of different laws and regulations, the lack of trust among accreditors and the vital nature of laboratory data for issues affecting health and safety. What I learned at the forum is that the problems of duplicate accreditations and redundant requirements that I see in an independent analytical laboratory are similar to problems experienced by laboratories in government, even within the same agency.
Is this an issue for microscopists? I think it is. Some of us are already well familiar with accreditation programs affecting anyone who does analysis by light or electron microscopy for asbestos. Others are trying to figure out how to deal with ISO 9000, which is intended for organizations that produce goods and services, but not really focused on laboratory operations. The organizers of NACLA are working to develop a system which will have all laboratory accreditations traceable to a single, international standard, probably similar to the present ISO Guide 25. They are also trying to resolve the present conflicting roles of NIST in laboratory accreditation, where NIST is at the same time accrediting accreditors, operating an accreditation system and operating calibration laboratories which should be accredited. At the same time, they are working to make all accreditations traceable to the Federal government through NIST in order to make them acceptable internationally. Finally, they are working to create a system in which various accreditation agencies recognize each other, probably because the accreditors themselves have been accredited by a process of peer review.
NACLA is just beginning, but microscopists should be aware that it is coming and, along with it, an increasing probability that each of our laboratories will be going through some sort of a standardized accreditation process.
I'm not aware that the proposal to develop NACLA is available on the web, but it should be available from NIST. If there are any questions, I'd be happy to try to answer them.
Andy
Andrew W. Blackwood, Ph.D. Structure Probe, Inc. P.O. Box 656 West Chester, PA 19381-0656 Ph: 1 610 436 5400 FAX: 1 610 436 5755 e-mail: ablackwood-at-2spi.com WWW: http://www.2spi.com
Paul wrote: } Another possibility is to plasma clean the holder. Most of the } contamination resident on holders is organic (hydrocarbon). An air or } oxygen/argon plasma is quite effective in reducing this contamination. The } plasma creates disassociated oxygen which chemically combines with the } carbonaceous material and reduces it to CO, CO2 and H2O. Depending on the } ion energies used, cleaning can occur without adversely effecting the } specimen holder.
Can Be cups be cleaned in plasma systems without damage?
Larry wrote:
} Certainly, there should be no necessity for a regular cleaning routine and } in general, I have never cleaned holders for which I have had } responsibility. The only exceptions are the external O-ring seal on the } barrel and the jewel bearing on the tip.
Does your experience include AEM in high resolution FEG's? We are about to purchase a 300 keV FEG and would like to know how critical specimen and rod cleanliness is.
Ciao for now, Ken
Kenneth JT Livi Department of Earth and Planetary Sciences 34th and Charles Streets The Johns Hopkins University Baltimore, Maryland 21218
Jim Darley is right about the EDTA, we tried it many moons ago (memory is the first thing to go!!) but found 1500 NF units/ml ovine hyaluronidase in millipore filtered seawater was better, for spermatoza at least. (D. G. Atwood et al,1975, Journal of Microscopy, vol 103, pp. 259 to 264.) I'll send you a reprint if you like!
"Others have referred to mucous which may or may not be removed. EDTA fell out of favour as a decalcifier some time ago. For the same reasons I would prefer a broad spectrum enzyme. I used a snail enzyme many years ago for removing mucous, but your local biochemist should be able to advise."
Cheers :) George Department of Earth and Atmospheric Sciences University of Alberta Edmonton, Alberta T6G 2E3 Canada ph: 403-492-5746 fax: 403-492-2030
An issue which I have not seen mentioned in the responses to this thread is the video bandwidth of the SEM amplifiers. SEM's are not (in general) designed to obtain quality images at TV rates, and the amplifiers are not usually capable of passing information at a high enough rate. This is quite separate from the noise issue, and no amount of frame averaging can correct the problem.
A way you can see this is to turn the electron beam off, turn up the PMT voltage until you can see noise pulses, then grab a single frame at TV rate. If you look closely, you will see that the noise pulses are not spots, but short lines. As an approximation, you can never get finer horizontal detail in your image than the length of these lines.
The demonstrations you will see by board vendors showing how their boards clean up a noisy image are just fine, but remember that they are using high quality CCD video cameras as the image source, and high bandwidth amplifiers, with the noise artificially enhanced, for their demonstrations.
As people have commented, one can get a useful image by frame-averaging a TV rate signal, but (in most cases, at least) don't expect it to have the same quality as a slow-scan image.
Tony Garratt-Reed.
**************************************************** **************************************************** ** ** ** Anthony J. Garratt-Reed ** ** Room 13-1027 ** ** Center for Materials Science and Engineering ** ** Massachusetts Institute of Technology ** ** 77 Massachusetts Avenue ** ** Cambridge, Massachsetts 02139-4307 ** ** U. S. A. ** ** ** ** Phone: 617-253-4622 ** ** Fax: 617-258-5286 or 617-258-6478 ** ** ** **************************************************** ****************************************************
**************************************************** **************************************************** ** ** ** Anthony J. Garratt-Reed ** ** Room 13-1027 ** ** Center for Materials Science and Engineering ** ** Massachusetts Institute of Technology ** ** 77 Massachusetts Avenue ** ** Cambridge, Massachsetts 02139-4307 ** ** U. S. A. ** ** ** ** Phone: 617-253-4622 ** ** Fax: 617-258-5286 or 617-258-6478 ** ** ** **************************************************** ****************************************************
} Can Be cups be cleaned in plasma systems without damage? } Dear Ken, Yes. I'd be careful where the pump exhaust is vented, however. The presence of Be dust in the exhaust is a possibility (at least in theory), and that is *very* toxic. The particles would likely be in the submicron range, and therefore easily inhaled. I'd think it unlikely that a Be cup with a smooth surface would be etched too readily. Does anyone else on the list have other info? } } Does your experience include AEM in high resolution FEG's? We are about to } purchase a 300 keV FEG and would like to know how critical specimen and rod } cleanliness is. } No, I have no experience with FEG instruments. I'd think the limiting factor would be how a dirty rod affects the vacuum. If you are interested in looking at specimens which have clean surfaces, the contamination would also be a major concern. Yours, Bill Tivol
A nice resource for checking full journal titles from standard abbreviations can be found at Chemical Abstracts WEbsite: ...its also nice cause they have the CODENs in the list and with Netscape or Internet explorer, one can type "command-f" to find the abbreviation of interest. This has all CAS-journals, which is most of the good ones... its at:
http://info.cas.org/sent.html
I recommend bookmarking it..
as Martha might say "it's a good thing."
Brendan Foran
_____________________________________________________________________ Brendan J. Foran Ph.D. ...currently just a "Post-Doc"
Dept. of Materials Science & Engineering 2125 H.H. Dow building The University of Michigan phone:(313)-763-4196 2300 Hayward Street fax: (313)-763-4788 Ann Arbor, MI 48109-2136 bforan-at-umich.edu _____________________________________________________________________
--IMA.Boundary.561329258 Content-Type: text/plain; charset=US-ASCII Content-Transfer-Encoding: 7bit Content-Description: cc:Mail note part
Bill Mason responded to an inquiry regarding Kodak digital cameras:
We have used a number of digital cameras, including Kodak. The Kodak cameras are not really so suitable for image analysis of the type you mention because they are really on 8 bit cameras as standard (256 grey levels), so have limited dynamic range.
I though that the following from Kodak's Readme file that comes with the latest driver download might be helpful especially since it seems to say that the data for their DCS 420/460 cameras are12 bit images:
"When the "12 bit Acquire" checkbox on the driver is checked, the driver will acquire the image data into Photoshop Version 2.5 or higher with 12-bit resolution per color. The file size is doubled and the acquire time is slightly longer. If the "12 bit Acquire" checkbox is unchecked, the acquire module passes 8-bit image data back to Photoshop.
Kodak's DCS cameras capture image data with a 12-bit analog to digital converter. Photoshop supports a 16-bit per pixel mode, however Kodak calls the new driver feature "12 bit acquire" so that we will never mislead customers into thinking that we use a 16-bit analog to digital converter.
Our DCS drivers that run on Macs and PCs maintain this 12-bit resolution through the image processing path. Early versions of Photoshop required acquire modules to reduce the image data resolution to 8-bits per pixel per color just before passing the image data back to Photoshop. Photoshop Version 2.5 or above allows acquire modules to provide the image data with either 8 or 16-bits per pixel per color. Adobe calls these 24-bit and 48-bit RGB Color modes." Any and all copyright notices may apply to the preceeding excerpt.
I have no financial interest in nor am I recommending use of Kodak,Photoshop,Mac etc.
Received: from deliverance.acsu.buffalo.edu (128.205.7.57) by ccmailgw.mcgawpark.baxter.com with SMTP (IMA Internet Exchange 2.03 (Beta 5) Enterprise) id 0005047D; Fri, 10 Jan 97 11:54:19 -0600 Received: (qmail 3190 invoked from network); 10 Jan 1997 17:55:29 -0000 Received: from listserv.acsu.buffalo.edu (128.205.7.35) by deliverance.acsu.buffalo.edu with SMTP; 10 Jan 1997 17:55:29 -0000 Received: from LISTSERV.ACSU.BUFFALO.EDU by LISTSERV.ACSU.BUFFALO.EDU (LISTSERV-TCP/IP release 1.8c) with spool id 7347505 for CONFOCAL-at-LISTSERV.ACSU.BUFFALO.EDU; Fri, 10 Jan 1997 12:55:21 -0500 Received: (qmail 222 invoked from network); 10 Jan 1997 17:45:19 -0000 Received: from arcs.cc.bbsrc.ac.uk (149.155.100.5) by listserv.acsu.buffalo.edu with SMTP; 10 Jan 1997 17:45:19 -0000 Received: from pserv.iapc.bbsrc.ac.uk by arcs.cc.bbsrc.ac.uk with SMTP (PP); Fri, 10 Jan 1997 17:47:59 +0000 Received: from mserv.iapc.bbsrc.ac.uk by pserv.iapc.bbsrc.ac.uk; (5.65/1.1.8.2/16Nov95-0425PM) id AA27505; Fri, 10 Jan 1997 17:45:16 GMT Received: by mserv.iapc.bbsrc.ac.uk; (5.65/1.1.8.2/20Mar96-1052AM) id AA03023; Fri, 10 Jan 1997 17:45:46 GMT Alternate-Recipient: allowed Auto-Forwarded: prohibited Content-Return: allowed Disclose-Recipients: prohibited Conversion: allowed Importance: normal Priority: normal Sensitivity: Company-Confidential X-Mailer: MAILworks 1.7-A-2 Message-ID: {970110174543.455-at-mserv.iapc.bbsrc.ac.uk.0} Newsgroups: bit.listserv.confocal
Dear MSA listserver subscribers, I am a graduate student (master's) trained in EM. I was wondering if anyone could tell me what range of starting yearly salary I should expect.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Colleagues:
I have read, with interest, all of the discussions concerning cleaning of TEM specimen holders. As has been mentioned, plasma cleaning has been determined to be a highly effective means to remove organic contaminants from both a specimen and a specimen holder. Significant work has been done in quanitfying the effects of Plasma Cleaning since Dr. Nestor Zaluzec at Argonne National Lab received his patent on the technique.
I am quite sure that there will be some lively discussions at the MRS Meeting in San Francisco during the TEM Specimen Preparation Symposium. If you'd like to get a jump start on the discussion, I can refer you to two articles on Plasma Cleaning that may be of interest:
1) "Simultaneous Specimen & Stage Cleaning for Analytical Electron Microscopy", Microscopy Today October 1996. Volume 96-8, Page 16, by yours truly. This is a very general discussion of the process.
2) "Reactive Gas Plasma Specimen Processing for Use in Microanalysis & Imaging in Analytical Electron Microscopy" by Nestor Zaluzec. This is a preprint of a paper to be presented at MSA in Cleveland this coming August. This paper provides parameters for processing and gives quantified data concerning the effects of plasma cleaning.
I can provide you with copies of both papers if you have an interest. You may also be interested in reading Dr. Zaluzec's patent (#5,510,624). I can also send you a copy of that if it is of interest.
If you have an interest in subscribing (IT'S FREE!) to Microscopy Today, you should send an e-mail request to Don Grimes at MicroToday-at-aol.com. It is a fine publication and I highly recommend it to all microscopists. I have no financial interest in Microscopy Today - I'm just a faithful reader!
DISCLOSURE I must also point out that South Bay Technology does license this tachnology from Argonne National Lab pursuant to the above mentioned patent. We also produce a plasma cleaner which is designed to clean specimens and holders so we have a vested interest in making you aware of the technique.
Please address your requests to my attention.
Best regards-
David Henriks TEL: 800-728-2233 (toll-free in USA) South Bay Technology, Inc. 714-492-2600 1120 Via Callejon FAX: 714-492-1499 San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com http://www.southbaytech.com
Manufacturers of Precision Sample Preparation Equipment and Supplies for Metallography, Crystallography and Electron Microscopy.
} Larry wrote: } } } Certainly, there should be no necessity for a regular cleaning routine and } } in general, I have never cleaned holders for which I have had } } responsibility. The only exceptions are the external O-ring seal on the } } barrel and the jewel bearing on the tip. } } Does your experience include AEM in high resolution FEG's? We are about to } purchase a 300 keV FEG and would like to know how critical specimen and rod } cleanliness is. } } Ciao for now, } Ken } } Kenneth JT Livi } Department of Earth and Planetary Sciences } 34th and Charles Streets } The Johns Hopkins University } Baltimore, Maryland 21218 } } klivi-at-jhu.edu (e-mail)
Kenneth,
I have done a lot of AEM, up to 300kV although not on FEGs. However, much of this has included EELS which will pick up C contamination more easily than any other method. Specimen rod cleanliness is important but I guess the point I was trying to make is that you shouldn't get it dirty in the first place! In general, and with proper care I believe that cleaning the specimen rod is unnecessary.
I assume you are not talking about UHV at the specimen - if you are, then the whole question of cleanliness is a different order of magnitude. If you are talking about UHV at the specimen, then probably Nestor is the person to comment on this.
Specimen cleanliness is a somewhat different issue and there are certainly some specimen prep procedures, and types of specimen that can lead to contamination problems. For example, there are some pretty gungey mixes around for electrochemical thinning - I used to put plenty of glycerol into one of my favourite mixes for stainless steel - these need carefully removing from specimens. Also, dirty ion beam thinners make pretty good fine grain carbon coaters. Be careful.
} At 08:48 10.1.1997 +0000, you wrote: } } } If it is an EDX detector, I would suggest that if you are really concerned, } } you should have a routine for users to disconnect the HT from the detector. } } Then if the detector should run dry, there is no risk of damage (although, } } Hello Larry, } } I would not give people the idea that you can let an EDS detector warm up } while connected into the microscope. This is true especially with detectors } with window. The dewar is pumped down to its ultimate vacuum by a chemical } sorption pump. If you let the detector warm up the pressure in the vacuum of } the dewar gets worse than that inside the microscope. The window fitting } does not normally like this direction of pressure difference and the result } may be your window (Be or polymer) blowing out into the microscope. At least } Tracor recommends to remove the detector before warming it up and again } cooling it down before installing to the microscope. } } Regards, } Jouko
I certainly wouldn't suggest anyone try warming up EDX detectors, on or off the microscope, without first talking to the manufacturer regarding warming/cooling procedures. However, from personal experience of Be window detectors, they can survive warming up while on a TEM column, so if it does happen to you (and you are responsible), try cooling it down again, before you cut your throat :)
With UTW detectors, you probably won'tt be so lucky.
Regards, Larry Stoter
--------------------------------------------------------------- Dr L. P. Stoter Technical Editor, MICROSCOPY & ANALYSIS Technesis 17, Rocks Park Road email: LPS-at-teknesis.demon.co.uk Uckfield, E. Sussex Phone: +44 (0)1825 766911 TN22 2AT Fax: +44 (0)1825 766911 United Kingdom ---------------------------------------------------------------
Thanks for all the pointers to Philosophical Magazine. Much appreciated.
Unfortunately, I'd already discovered the highly obscure publication Philosophy of Magic. After the obligatory puff of greasy black smoke, I find myself in the form of a large green frog. You have no idea how complicated it is typing this with webbed fingers...
On the bright side, my debugging skills seem to have improved markedly.
VBS is the only automatic LN2 filling system I have seen commercially, although there may be others. However, you might want to design your own system, using cryogenic solenoid valves and sensors, and providing LN2 from cylinders. It is fairly inexpensive to do this. I can send you information on a system we built and used in our lab for some time, if you are interested. I presented a poster on this system at the 1994 MSA meeting and the description in included in the Proceedings from that meeting. One problem we ran into with our system is that we did not have a failsafe mechanism to shut the system off when the cylinder ran out of liquid. The result was that nitrogen gas would continue to fill the dewar and eventually would blow out the remaining liquid. I think this can be remedied by making some changes to the system. Regards, Melanie A. Behrens Texaco, Inc. P.O. Box 509 Beacon, NY 12508 914-838-7261 behrema -at- Texaco.com or MelanieOwl -at- aol.com
Dear Gerry, I have looked at the growth rings in human (child's) teeth, and the dark marks left by early stress events. I felt that light microscopy gave a better view of the dark marks, but examining the teeth was not difficult. I just gold-coated and examined as usual. I did not try to determine any ratios. I do believe that there is some shrinkage, due to dehydration and cryo would be a more rigorous way to go. Good luck, Mary At 02:42 PM 1/10/97 +1000, you wrote:
} Would any of you kind electron microscopists have looked at the growth } rings in seal teeth or any mammalian teeth, please? And how did you do } that? And is there a relationship between the ratio of cementum and dentine } to the age of the mammal? } Thank you kindly } Cheers } Gerry nash Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
Dear Fellow Microscopists, I'm looking for a full time position in a research or imaging= facility lab. =20 I have a BS in Biology and I'm currently persuing a second BS in biomedical= photography with a concentration on photomicrography, and digital imaging. = I'll be graduating on February. You can view my resume at http://www.isc.= rit.edu/~caa3045
My photography, computer skills, research and laboratory experience is= varied. During the summer of 1992, I conducted research on the effects of= hexadecane on genetic variability of nestmate recognition in honey bees at= the University of Colorado at Boulder. During the summer of 1993, as part= of a research program sponsored by the Medical College of Pennsylvania, I= characterized chloride channels in the human colonic cell line T84 using= the patch-clamp technique.=20
My research technician position from January '94 thru June '95 at the= Medical College of Pennsylvania expanded my range of skills. I conducted= experiments on the structural change of F-actin and ankyrin in the= cytoskeleton of lymphocyte using the confocal microscope and various immuno= cytochemistry techniques. The result of this project has already been= published in the journal of Membrane Biology 147, 283-294. In addition, I= used the patch clamp technique to investigate bile duct cells from Cystic= Fibrosis patients, and characterize fibroblast cells (NIH3T3). =20 Also, my position at the Ocular Cell Transplatation Laboratory at the= University of Medcine and Dentistry of New Jersey involved digitizing and= capturing images of the retina, create plates for publication, and created= and maitained the laboratory's website. I am presently the webmaster for = Spectra Services. =20
In addition to my science background the Biomedical Photographic= Communications Department at RIT has exposed me to many computer skills = and medical photography techniques. I have hands on experience on fundus= photography, stereo photography, and I am familiar with the fluorescein= angiogram techniques. In addition, I am experience with many formats of= films processes, black-and-white, and color printing, photomacrography,= photomicrography, darkfield, brightfield, nomarski, phase-contrast,= polarizing, Rheinber, SEM,fluorescence, and confocal microscopy. =20
Also, I have a great interest and experience in digital photography,= multimedia, and computer in general. I have hand on experience with the= window platforms and Macintosh computer (Macintosh major area of strenght).= I am proficient in many imaging and multimedia production softwares such= as photoshop and Multimedia Director, QuarkXpress, Adobe Illustrator, and = Adobe Premiere. In addition I have work experience with HTML and Lingo= programming. =20
My career objective is to become an expert in a wide variety of imaging= enhancing, analysis softwares and equipments and become very= knowledgeable in the area of microscopy (Confocal, TEM and SEM.) And create= interesting interactive multimedia pieces conveying scientific= informations. If you have access to the internet you can view my resume= and some of my images in my web site at http://www.isc.rit.edu/~caa3045 =20 If you think I may contribute to your department or if you need= further information please send me an e-mail to "caa3045-at-rit.edu" or "alm= onte-at-medcolpa.edu" Thanks,
--Ciprian Have fun and keep the sun on your back and a smile on your face. __________________________________________________________ Ciprian A. Almonte Rochester Institue of Technology Biomedical Photographic Communications Rochester, NY 14623-5603
Visit my web site at http://www.isc.rit.edu/~caa3045/ __________________________________________________________
The UC Berkeley Electron Microscope Lab is looking for and EM Tech. What follows is the job announcement.
Staff Research Associate II (PSS 1) Electron Microscope Laboratory $30,200/ year starting salary Job number: 12-411-30/SL Closing date: 1/31/97
Operate and maintain a TEM, Bal-Tec High Pressure Freezing Machine, and JEOL 9000 Freeze-fracture machine. Train students, staff, and other users in TEM technique, including sample preparation. Operate cryofixation equipment for EM sample preparation. Train users in darkroom technique. Learn and apply new techniques as appropriate.
Qualifications: Experience in electron microscopy, especially TEM sample preparation techniques and cryotechniques. Effective communication skills. Experience with TEMs and related equipment. Experience with specimen preparation techniques for TEM. General Lab skills such as preparing buffer solutions and opreating pH meters. Experience with electronic equipment and its routine maintenance. Darkroom experience. Ability to work independently.
If you are interested please contact the UC Berkeley Employment Office.
UC Berkeley Employment Office Room 7-G (Ground Floor) 2200 University Avenue Berkeley, CA 94720 (510) 642-1011 general line (510) 643-9421 TTY for disabled
I have a specimen preparation problem which some of you on the list may be= able to help me with.
I have been using M-BOND 610 to prepare my TEM cross-sectional samples with= great success, but I am now worried that the elevated temperature curing= (200C for two hours) is annealing the thin films I am examining(I am= looking at copper thin films on a quartz substrate, which I sandwich= between wafers of silicon for the cross sections). Has anyone had any good= experiences using a room temperature adhesive for cross-sections which will= be tripod polished and briefly ion milled?
F. Scott Miller Electron Microscope Lab smiller-at-umr.edu University of Missouri-Rolla =20 223 McNutt Hall voice: 573 341 4727 Rolla, MO 65409 USA fax: 573 341 6934
In my ~20+ years of experience, all specimens contaminate to some degree (some slow, some fast) in the microscope. I've seen this in every instrument (TEM, STEM, SEM) I have used from W, LaB6 to FEG instruments, UHV or not. In modern TEM/STEM instruments the majority of this contamination is specimen and stage borne as the manufacturers have generally improved their vacuum systems over the years so that their contribution is small to neglegible.
In the specimen borne regime, the magnitude of the contamination is a function of the sample (metallic, semiconductor, organic, ....) , it's preparation method (electrochemical, chemical, microtoming, ion milling...), the microscope, the probe and probe current. Plus a number of other less well controlled factors.
Rather than draw this out into a very long dicussion, it suffices to say that when critical small probe work is being done, I plasma process my specimens and stage before microanalysis especially in LaB6 and FEG systems. Or if I determine after an experiment starts that the specimen is begining to contaminate, then I remove the sample and the stage from the microscope and process them off-line and then resume to work. The effectiveness of this processing depends upon the gas, pressure, and power of the plasma but is dramatic in most situations. I have been able to minimize/remove specimen borne contamination to a level where I can operate for ~ 8+ hours without significant contamination. Of course, when it reappears (usually now due to the microscope) the specimen is just "reprocessed" and I can continue working. Unfortunately, once surface hydrocarbons are removed you may find out that your sample exhibits electron sputtering in the microscope :-( . This is an effect which we calculated and showed would happen if the conditions are right back in 87 (Zaluzec & Mansfield in Proc. AEM-87). Adding back a very thin layer of spectroscopically pure carbon sometimes cures this problem, with minimal contamination effects.
I also routine apply this process to stages which have Be cups. If you operate under the correct plasma conditions you will NOT sputter/etch material from or onto the stage. This only occurs when the power level of the plasma is too high and you enter the etching or "ashing" regime.
Generally I would recommend a power level of ~ 5-10 W, pressure ~200 mT, processing time ~ 5 min and a 2 stage cleaning. First using pure Argon, followed by pure Oxygen. I have experimentally found that this always produces the best results. In addition, the temperature rise under these conditions is less than 5 C, so specimen/stage heating is almost never a problem.
As per the rules of the Microscopy Listserver. I should point out that all the commerical suppliers (SPI, SBT, EAF) of TEM specimen/stage plasma cleaning technology are licensees of a US Patent, which was issued to my employer Argonne National Lab and ANL obviously has some financial interest in that patent and this methodology.
SCOTT, TURN THAT OVEN DOWN! (Pardon me, I don't often shout.)
Curing M-Bond 610 at 200C for 2 hours is waaaayyyyyy overkill. The instruction bulletin that comes with M-Bond suggests 125 to 160C for two hours, and remember that's for bonding strain gauges to steam boilers!
We never exceed 70C. If we are gluing a specimen on to a grid by clamping the specimen in self-closing tweezers and then inserting tweezers+grid in an oven we cure for 2 hours. (If the glue is still soft (rare) we'll add another hour). If we are curing the specimen to a grid on a glass slide on a hot plate at 70C we find 10 to 15 minutes to be adequate. Remember to put a pan of water in the oven to keep the humidity up during curing!
For temperature sensitive parts we have experienced no problems curing at 30, 40, etc degrees up to 70C. Room temperature curing M-Bond takes overnight. Paul Albarede, France's premiere tripod polisher, routinely cures M-Bond overnight at room temperature, he tells me--arguing that the differential contraction of the Cu grid and the specimen leads to stress being put on the thin specimen when the Cu grid and specimen cool off after bonding at temperature. You can see this with Si if you cure at too high a temperature: the flat polished specimen ends up with a wavy edge like curly lasagna pasta.
NIST-NIH Desktop Spectrum Analyzer (DTSA) Spring 1997 Workshop
A three-day intensive Workshop on NIST-NIH Desktop Spectrum Analyzer (DTSA) will be held at NIST, Gaithersburg, Maryland, March 26-28, 1997. DTSA is a software platform for electron-excited energy-dispersive and wavelength-dispersive X-ray spectrometry developed by Fiori, Swyt, and Myklebust for Macintosh computers. The Workshop will cover practical aspects in utilizing DTSA, including spectrum processing (background and peak interference removal), quantitative analysis for bulk and layered specimens (matrix corrections, including the comprehensive CIT-ZAF resource), analytical electron microscope quantitation for thin foils (Cliff-Lorimer sensitivity factor method), generation of X-ray spectra from first principles, and development of analytical strategy through "desktop" simulations. Attendance at the DTSA Workshop is free, but is limited to 25 attendees.
For reservations and/or information, contact Dale Newbury at dale.newbury-at-nist.gov or telephone 301-975-3921. fax 301-417-1321
Note: DTSA is a software program sold through NIST's Standard Reference Data Program and we therefore have a very small financial interest in the product.
Eric B. Steel, Leader e-mail: eric.steel-at-nist.gov Microanalysis Research Group Office: 301-975-3902 N.I.S.T. FAX: 301-216-1134 Bldg. 222/Rm A113 Gaithersburg MD 20899
Please post the following advertisement for an academic position at The University of Calgary for a Structural Biologist/Microscopist.
STRUCTURAL BIOLOGIST
The University of Calgary Department of Anatomy invites applications for a fulltime academic position as a structural/cell biologist. This position offers an excellent opportunity for independent and collaborative research with other structural biologists employing technologies such as magnetic resonance spectroscopy and x-ray diffraction in the university-wide structural biology group, and for access to the University s Microscopy and Imaging Facility, comprising state-of-the-art electron and computer-based light microscopies. Duties will also include undergraduate teaching and graduate student supervision.
Qualifications include a PhD or equivalent, at least two years of postdoctoral experience, and a proven record of excellence in a research program which includes the development of advanced imaging techniques. Researchers particularly encouraged to apply are those with interests at the cellular or molecular level in an area complementing activities of a Faculty of Medicine research group such as Cancer Biology, Molecular & Developmental Biology, Joint Injury & Arthritis, etc. More information is available at http:/www.ucalgary.ca/~resoff/index.html.
The successful candidate must compete successfully for salary and establishment grant support from the Alberta Heritage Foundation for Medical Research and/or the Medical Research Council, and will have 75% of time protected for research.
In accordance with Canadian immigration requirements, priority will be given to Canadian citizens and permanent residents of Canada. The University of Calgary is committed to Employment Equity.
Please submit a curriculum vitae and statement of research and goals, and arrange for three letters of reference to be sent directly, by February 15, 1997, to:
Dr. D.P. Bazett-Jones Department of Anatomy The University of Calgary 3330 Hospital Drive N.W. Calgary, Alberta, Canada T2N 4N1
Tony King of VG Microtech in the UK (who produce a cryo-SEM, the Polaron LT7400) had some additional comments which he asked me to pass along:
} } } My recommendation would be to use a mixture of Tissue-Tek and carbon } } } dust (from carbon rod sharpener) paste on your cryo-stub. In this } } } mixture the eggs will not sink fast, therefore will have time to carry out } } } the freezing process.
Tony replies: Try tissue tek smear (with carbon dust on a Dry colloidal graphite base; this absorbs the tissue tek bulk and holds the samples in place.
} } } I have great LTSEM results using this mixture with unfixed single cell } } } culture and bacteria. The separate problem about the fine layer of salt, } } } (if the osmatic change is all right!) use larger amount of distilled water } } } to rinse for longer time.
Tony replies: } Use Osmium vapour to semi-fix the cells before plunging into water. } Osmotic shock is then reduced to minimum.
} Regards, } } Tony King } Product specialist } VG Microtech/ Polaron range } } Tel: +44 (0)1825 746251 } Fax: +44 (0)1825 768343 } } E&OE }
Best regards, steven E. Slap ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
Good afternoon, I've been asked by a student about possible fixation protocols for the analysis of Spisula solidissima (surf clam) larval development by SEM. The protocol she has been using has resulted in considerable shrinkage. She is fixing the material in 4% paraformaldehyde in buffered "sea water". She was unable to give me the exact reference at the time, but said it was by Longo. Any and all input would be greatly appreciated.
Thanks in advance.
Dwight Beebe E-mail: beebed-at-ere.umontreal.ca Institut de recherche en biologie vegetale Voice: 514-872-4563 Universite de Montreal FAX: 514-872-9406 4101, rue Sherbrooke est Montreal, Quebec H1X 2B2 Canada "Time flies like an arrow; fruit flies like a banana"
I am looking for a stain called "Crossmon trichome" (this may have been misprinted in the reference). Can anyone help me locate a supplier? I have tried the obvious big chemical supply companies.
To the Microscopy List, I've got some 30 um thick frozen dog pituitary sections which are showing punctate autofluorescent staining (in both Fitc and Rho channels), without antibodies. Does anyone have suggesstions on reducing this annoying autofluorescence, specific concentrations, time and temps would be appreciated.
Dear Scott, I have used high-strength epoxy (24 hour, not 5 minute hardening time) to prepare cross-sections of ion-implanted Gallium Arsenide. We were waiting for the M-610 to be delivered. It is slow but does not raise the temperature much in the thin layers. I had the shop make a small, parallel-jawed vise with teflon-lined jaws to clamp the sample in. These samples were dimpled, then ion-milled. At 12:47 PM 1/12/97 -0600, you wrote:
} I have a specimen preparation problem which some of you on the list may be able to help me with. } } I have been using M-BOND 610 to prepare my TEM cross-sectional samples with great success, but I am now worried that the elevated temperature curing (200C for two hours) is annealing the thin films I am examining(I am looking at copper thin films on a quartz substrate, which I sandwich between wafers of silicon for the cross sections). Has anyone had any good experiences using a room temperature adhesive for cross-sections which will be tripod polished and briefly ion milled? } } } F. Scott Miller Regards, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
1/97 SUMMER MICROSCOPY TECHNICIAN POSITION AVAILABLE
A three month (June, July, and August) position is open for a microscopy oriented technician at the Marine Biological Laboratory, Woods Hole, MA. We would like to attract someone with some knowledge of biological preparative techniques and experience in laser scanning confocal microscopy, TEM, SEM, and/or LM.
The technician will assist in the Central Microscopy Facility. The technician's duties will be to check out incoming investigators in the usage of our equipment and then to supervise its continuing usage and to perform contract work for investigators. This may include fixation, embedding, sectioning, scope use, darkroom work, etc. The technician will also provide routine maintenance.
This is a short term and scientifically rewarding position. Salary will be in the $7 to $9/hour range. Housing may be available to rent through MBL.
For more information, including a more detailed position description, please contact Louis Kerr at: MBL, 7 MBL Street, Woods Hole, MA 02543. Telephone, 508-289-7273; or at Email: lkerr-at-mbl.edu. Please apply to: Human Resources, MBL, 7 MBL Street, Woods Hole, MA 02543. or resume-at-mbl.edu.
An Equal Opportunity/Affirmative Action Employer/ Non-smoking workplace.
Louie Kerr Research and Education Support Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543 508-289-7273 508-540-6902 (FAX)
We are looking for a powerful software to catalog, store and reteive images. We prefer something that is compatible with both PC and the MAC. Any suggestions?
__________________________________________ Rex A. Hess, Ph.D., Associate Professor, Director, Center for Microscopy and Imaging University of Illinois, Veterinary Biosciences, 2001 S. Lincoln, Urbana, IL 61802-6199 217-333-8933; FAX 217-244-1652; email: r-hess-at-uiuc.edu homepage-- http://www.cvm.uiuc.edu/HomePages/rhess/hesspage.html
We are contemplating digitizing our darkroom and seek any thoughts you might provide on scanners and printers. This is primarily an EM lab so we would be scanning standard 3.25" X 4.0" EM negs. and 4" X 5" Polaroid SEM negs.
We are considering multiformat film scanners, specifically the Nikon LS-4500AF and the Polaroid Sprintscan 45, and flatbed scanners, specifically the Microtek ScanMaker III and the UMax PowerLook 11.
Do you have any recommendations for neg. scanner vs flatbed scanner? Of the models listed, does anyone have any experience - good or bad?
We have also thought about the Leaf MicroLumina and understand that it is excellent. Is the MicroLumina's performance worth the price differential?
As far as a printer, we're basically set for dye-sub. printers but would like to get a laser printer for "routines". I've pretty much narrowed it down to the Lexmark OptraR plus with about 32 meg. of memory. Does this sound reasonable? Got any other recommendations.
Any input would be greatly appreciated. TIA for your help.
John
John G. Aghajanian, Ph.D. Worcester Foundation for Biomedical Research 222 Maple Avenue Shrewsbury, MA 01545 Phone: 508 842-8921 ext. 147 Fax: 508 842-9632 email: johna-at-sci.wfbr.edu
This is a rather important process that should be done very carefully. I devoted several pages to discussing methods for cleaning parts for vacuum systems in my book 'Vacuum Methods in Electron Microscopy' p.69-74. If you are using the standard top-entry type of holder, cleaning should be straightforward - scrub it thoroughly with Tilex Soap Scum Remover, rinse with hot running water, sonicate in a strong detergent solution, rinse with hot tap water, rinse with reagent grade isopropyl alcohol, dry with a gas blaster. If you are using a side entry stage you can use essentially the same procedure, but you must then be careful to avoid getting the solutions inside the holder if it is one that has provisions for manipulating the specimen. Often, enough cleaning can be done to get rid most contamination problems by sonicating just the end of the holder in isopropyl alcohol, then drying with a blaster. The latest method for these holders is Plasma Discharge Cleaning, and Southbay Technologies markets a device that is specially designed for this purpose. W. C. Bigelow (bigelow-at-umich.edu)
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
There are many reasons for NOT using grease-base polishes for cleaning parts to be inserted into the interior of high vacuum sysstems (See Vacuum Methods in Electron Microscopy, Portland Press, pp. 69-74). Basically, this is the equivalent of taking a bath in a mud puddle. One principal reason for cleaning is to remove hydrocarbon materials from the surfaces of the parts, and so it makes no sense at all to use a greasey material to do the job. In addition, as noted by others, the grease and abrasive materials are likely to get embedded in cracks and crevice and then not be completely removed, whereupon they will act as a very effective source of contamination. Very effective cleaning can usually be accomplished simply by thoroughly scrubbing with one of the many modern detergent solutions formulated for use in the electronics inductry (see above reference) or with Tilex Soap Scum Remover (available in most supermarkets), rinsing with running hot tap water, ultra sonic treatment in a warm detergent solution, rinsing again with hot tap water, rinsing with reagent grade isopropyl alcohol, and drying with a gas blaster. If you find you need an abrasive in the initial stage to remove stubborn deposits (or if you feel you must enhance the surface finish) try using a bit of Comet Cleaner (the kind formulated for use on plastic tubs and showers, which wont seriously scratch most metals) and then rinsing with hot water, before the initial scrubbing step. This procedure involves no solvents other than water and isopropyl alcohol (a common constituent of rubbing alcohol, and therefor perfectly safe to use) and so no expensive or complicated safety procedures are necessary, and it usually does the job quite nicely. The Tilex Soap Sum Remover will even remove silicone oils from most metal surfaces, and I have also used it to remove spots of various kinds from clothing, grease spots from carpets and auto seat covers, and semi-dried paint from my hands after painting. Needless to say, it works great for its intended purpose of cleaning bathtubs, wash basins, shower curtains and shower tiles. (No commercial interest, it is just very handy stuff to know about) W. C. Bigelow (bigelow-at-umich.edu)
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
I'm looking for a used EDX detector to interface to a Hitachi 2500 SEM, to use while upgrading the original Kevex Delta 3 Quantum system. Geometry is more important than make or model, but Kevex with or without thin window would be best. Please e-mail me with any possibilities and price.
A colleague asked if I could do EDXS for calcium deposits on the surface and/or in cells of histosections that are mounted, stained and coverslipped. It would mean removing the coverslip and mounting medium, and then get the surface really clean without loss of the region of interest (no pun intended). Is this the method to use or is there a better one and what would be the specifics of the method. Has anyone done this before? All I know is that the sections were stained with some sort of silver-type stain that is nonspecific for calcium but if present will show up as a dark body. The sections were not counterstained. Is there too much calcium in the glass that I wouldn't be able to see the particles against the background?
Thanks for any suggestions, they will be most appreciated.
A colleague here needs a used EDS detector, preferably Kevex, to fit a = Hitachi 2500 SEM. He tells me those that fit the Hitachi 2300-2400 models might also work. I = also need one or both of the video boards for a Kevex 8000 analyzer. = Anyone have some spares?
*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.* James J. McGee (jmcgee-at-sc.edu) Dept. of Geological Sciences University of South Carolina (803) 777-6300 (Office) Columbia, SC 29208 (803) 777-6610 (Fax)
We are looking for a powerful software to catalog, store and reteive images. We prefer something that is compatible with both PC and the MAC. Any suggestions?
__________________________________________ Rex A. Hess, Ph.D., Associate Professor, Director, Center for Microscopy and Imaging University of Illinois, Veterinary Biosciences, 2001 S. Lincoln, Urbana, IL 61802-6199 217-333-8933; FAX 217-244-1652; email: r-hess-at-uiuc.edu homepage-- http://www.cvm.uiuc.edu/HomePages/rhess/hesspage.html
----- End Included Message -----
Rex,
I use Aldus Fetch as a cataloging program for my Power PC. The images can be worked on with Photoshop on either PC or Mac. Image transfer is usually accomplished by ftp from one computer to another over a network. Others whom I know use it also like it.
Ed Basgall, PhD Res Assoc Dept. of Chemistry Surface Analysis Group Penn State Univ. 181 Materials Res. Inst. Bldg University Park, PA 16802
} I've been asked by a student about possible fixation protocols for } the analysis of Spisula solidissima (surf clam) larval development by SEM. } The protocol she has been using has resulted in considerable shrinkage. } She is fixing the material in 4% paraformaldehyde in buffered "sea water". } She was unable to give me the exact reference at the time, but said it was } by Longo. Any and all input would be greatly appreciated. } } Thanks in advance. } } } Dwight Beebe E-mail: beebed-at-ere.umontreal.ca
Have you tried (all together now) HMDS? [Hexamethyldisilizane]. I've had success with this with nudibranch veligers. Use a: 100% EtOH (3X washed)=} 3:1-1:1-1:3=} 100% HMDS (3X wash) change, drain off excess leaving specimens covered, and dry at 60 C. Do *everything* in a fume hood! But then soft little zooplankters like to shrink anyway. If you have access to one, you might be better off looking at fixed or fresh, hydrated specimens in an ESEM or variable-pressure scope tricked up to suck in water vapor; frozen hydrated specimens in a cryoSEM would be the other way to go. Phil
&&& Illigitimi non carborundum &&&&&&&& Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217)244-3145 days (217)355-3145 evenings oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
I hope you can post this postdoc position. Nina Allen
CALCIUM SIGNALING AND GRAVIPERCEPTION IN PLANTS
NCSU-NSCORT Postdoctoral Fellowships
Cell Biology-Imaging: POSTDOCTORAL POSITION is available immediately for the NASA Specialized Center of Research and Training (NSCORT) in gravitational biology. This is an opportunity to join a dynamic group of 12 project leaders and 5 postdocs studying the effects of altering calcium homeostasis on plant responses to gravity. The group is taking an integrated approach involving molecular, cellular, biochemical and physiological techniques. The specific project involves monitoring early changes in cellular calcium either by calcium ratio imaging and/or electrophysiology. Applicants do not require prior experience in plant biology but the calcium imaging applicant must have experience in calcium ratio imaging, image analysis, and microinjection. Electrophysiologists should have experience with either vibrating probes or patch clamping techniques as well as microinjection and computer analysis and be willing to collaborate with colleagues in the NSCORT as well as the National Vibrating Probe Facility at the Marine Biological Laboratory at Woods Hole, Massachusetts. Send curriculum vitae and names of three references to : Nina Stromgren Allen, Department of Botany, North Carolina State University, Raleigh, NC 27695-7612. Fax: 919-515-3436. Email: nina_allen-at-NCSU.EDU. NCSU is an Equal Opportunity Employer.
Nina Stromgren Allen Department of Botany Box 7612 North Carolina State University Raleigh, NC 27695-7612 Phone: 919-515-8382 (Office), 515-3525 (Lab), 515-2727 (Department Secretary) Fax: 919-515-3436
We are contemplating to buy a slow scan CCD camera for recording TEM CBED Kikuchi patterns and authomatically index them using Hough transform (amongst other things). This is to be part of a fully automated crystal orientation measurement facility, where "large" sample areas may be measured without user interaction.
The equipment is to be installed on a CM200. A 16 bit resolution is preferable (or eaven neccessary?). Are there anyone in the community who would like to share their experience on this matter with us? Any comments / advice will be grately welcomed.
Yours sincerely, Paul Baggethun.
============================= P.Baggethun (post-doc) Ecole des Mines de Saint-Etienne Centre SMS 158 Cours Fauriel F-42100 SAINT-ETIENNE, CEDEX 2 FRANCE =============================
Hi Folks, On the subject of image databasing raised by Rex Hess
We have had a lot of success using an inexpensive though very neat package called thumbs plus, you can down load it from any of the usual share ware or windows sites. It catalogs, does thumbnails, keywords, some basic image processing, works with a whole bunch of file formats, and in our case perhaps most importantly allows offline cataloging. we use CD's as our primary archive. This package provides a nifty offline catalog in which the CD title is seen which may be expanded to the entire directory structure together with thumbnails of the images. Older versions of the package got kind of slow with big databases (} 10K images) the current incarnation seems to remain pretty speedy even when loaded up with about 200 CDs plus continual updates of a large stack of networked drives. We are pretty happy with the package having tried several of the expensive, and inexpensive solutions
-- ======================================== Simon C. Watkins Ph.D. Associate Professor Director, Structural Biology Imaging Center Scaife 840 University of Pittsburgh Pittsburgh PA 15261
THis is a bit of an old saw but has anyone done a comparison of the currently available immersion oils wrt autofluorescence. Our light microscope platform is diverse (Zeiss, Nikon, Olympus) and slides etc travel from scope to scope which would lead to disastrous contamination problems if each 'scope had its own oil. We have been hanging on to Zeiss oil up to now , simply because we had a large bottle of the stuff. I have been a little unhappy with the levels of autofluorescence recently (prior to contamination ) and was looking for a change. Any ideas?
Simon
-- ======================================== Simon C. Watkins Ph.D. Associate Professor Director, Structural Biology Imaging Center Scaife 840 University of Pittsburgh Pittsburgh PA 15261
We are interested in buying a used TEM+EDX system. If you have, or know anyone who has one for sale, please send me a private email (we are willing to pay for the shipment).
Many thanks.
Redha Touaitia School of Engineering University of Northumbria at Newcastle Newcastle Upon Tyne NE1 8ST UK Tel : +44-191-227 36 14 email: r.touaitia-at-unn.ac.uk
I concur wholeheartedly. Thumbs Plus is fast, easy to learn, easy to use, flexible....buy it. Our R&D comuting department is considering the network-licensed version as a standard platform for image cataloging, in place of Microsoft Access and other high-power databases. AND for all you NIH Image users, a Mac version is planned.
Cerious Software can be found on the web at: http://cerious.catalogue.com/index.html
I have no financial or other interest (other than hoping the product continues to improve) in Cerious Software. ------------------------------------------------- Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 Phone: (508) 750-1717 E-mail: crossman-at-osi.sylvania.com
Our web sites: www.sylvania.com www.siemens.com --
Dear all, I have been asked to put the following job announcement on the listserver.
} Electron Microscopy Postdoctoral Position } Rutgers University } } The Ceramics Department at Rutgers University is seeking a postdoctoral } associate with an electron microscopy background. The candidate should be } interested in breaking new ground in the characterization of multicomponent } powder mixtures using a fully digital FE-SEM coupled with position-tagged } spectroscopy (PTS). PTS is a novel EDS method that can provide interaction } volumes as small as 50 nm and full spectrum scans within each pixel. The } candidate will work within a research group focused on understanding the } relationship between powder characteristics and the experimental and } theoretical achievable level of homogeneity in powder mixtures. Microscopy } work will be correlated with a state of the art mixedness simulation model } known as the concentric shell model of mixedness. The candidate should be } able to work within a team that has strong theoretical as well experimental } orientations. Candidates should demonstrate their ability to conduct } cutting-edge research in microscopy as well as effectively communicate with } others. The position is immediately available with a highly competitive } salary dependent on the candidate's qualifications and includes full health } care benefits. Candidates should submit their curriculum vitae, three } letters of reference, relevant publications and their anticipated starting } date by February 15, 1997 to: } } Richard E. Riman } Department of Ceramics } Rutgers, The State University of New Jersey } P.O. Box 909 } Piscataway, NJ 08855-0909 } } 908-445-4946 } 908-445-6264 (fax) } riman-at-erebus.rutgers.edu
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://sun1.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
MICROSCOPY & ANALYSIS is widely circulated among microscopists in the UK, Europe and the USA. Our editorial articles cover a broad spectrum of applications, techniques and instrumentation in all branches of microscopy and related analytical methods.
As I posted a few months back, we intend to start a "Questions and Answers" feature. Originally this was scheduled to appear in our January issue, but for various reasons was delayed. It will now appear in the March issue.
If you have any technical or scientific questions in the field of microscopy and related analysis that you would like to put to a large, expert readership, please e-mail them to me.
Regards,
--------------------------------------------------------------- Dr L. P. Stoter Technical Editor, MICROSCOPY & ANALYSIS Technesis 17, Rocks Park Road email: LPS-at-teknesis.demon.co.uk Uckfield, E. Sussex Phone: +44 (0)1825 766911 TN22 2AT Fax: +44 (0)1825 766911 United Kingdom ---------------------------------------------------------------
Add my vote to everyone elses regarding thumbs plus. Bang for buck, we have not found anything better.
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-bgsm.edu * **************************************************************
I am also in the search for an image archiving solution. I've played with several packages that work nicely as a "local" solution, but need to expand the scope to include accessability (and searchability) via our corporate intranet.
Specifically, is anyone aware of, or have experience with, an ODBC complient application which will run on an NT based server, that has an API to NETSCAPE? The local input application should be PC or MAC, and should handle multimedia as well as still images.
Thanks in advance,
**************************************** Don Steele Steele-at-KRDC.INT.Alcan.Ca Alcan International Kingston Research and Development Centre (613) 541 - 2145 ****************************************
I did do a test of immersion oil autofluor and found Nikon to be the lowest.
Bob Morphology Core U of Washington
On Wed, 15 Jan 1997, simon watkins wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } -- [ From: simon watkins * EMC.Ver #2.5.02 ] -- } } Hi Folks: } } THis is a bit of an old saw but has anyone done a comparison of the } currently available immersion oils wrt autofluorescence. Our light } microscope platform is diverse (Zeiss, Nikon, Olympus) and slides etc travel } from scope to scope which would lead to disastrous contamination problems if } each 'scope had its own oil. We have been hanging on to Zeiss oil up to now } , simply because we had a large bottle of the stuff. I have been a little } unhappy with the levels of autofluorescence recently (prior to contamination } ) and was looking for a change. Any ideas? } } Simon } } -- } ======================================== } Simon C. Watkins Ph.D. } Associate Professor } Director, Structural Biology Imaging Center } Scaife 840 } University of Pittsburgh } Pittsburgh PA 15261 } } tel: 412-648-3051 } fax: 412-648-8330 } ========================================= } } }
I concur with Simon's opinion below. I've been using a couple of versions of ThumbsPlus for over a year. In addition to making image databases, I use it to review my work at the end of the day. In combination with an Epson color ink jet printer (e.g. Stylus Pro or 500) I can perform a "sanity" check on my recent data in a very inexpensive manner.
} Return-Path: {Microscopy-request-at-Sparc5.Microscopy.Com} } Errors-To: Microscopy-request-at-Sparc5.Microscopy.Com } X-Sender: zaluzec-at-microscopy.com } Date: Wed, 15 Jan 1997 22:52:05 -0500 } To: microscopy-at-Sparc5.Microscopy.Com } From: simon watkins {swatkins-at-pop.pitt.edu} (by way of Nestor J. Zaluzec) } Subject: Re: catalog images } Errors-To: Microscopy-request-at-Sparc5.Microscopy.Com } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have had a few requests for a location for thumbs plus, you can download the demo from www.cerious.com
thanks
Simon
-- ------------------------------------------------- Simon C. Watkins Ph.D Associate Professor Director SBIC University of Pittsburgh Pittsburgh PA 15261 tel 412-648-3051 fax 412-648-2004 -----------------------------------------------
Probably a no-brainer :)...does anyone have any suggestions as to how I can stain agar blocks for embedding in LR White? So I can find them again in the final block? So far I've only tried Toluidine Blue - didn't work. I have some Coomassie-agar in the works right now, but it is clearing, too. Multiple suggestions are MORE than encouraged...because I'll ultimately need something that will stain the agar without messing up my antigens. Sigh. Thanks! Tamara CSHL, NY
We would like to know if anyone can suggest methods for preparing liposomes for SEM imaging other than those requiring freeze-fracture or other cold-stage techniques. Specifically, are there other methods for drying/preparation of a liposome suspension that can avoid dissolving the liposomes in the process. ******* ******* ******* A similar request was posted on this listserver by a Donna Turner {dturner-at-bcm.tmc.edu} on 12/20/96. The question was:
"I need information regarding processing for thin-sectioning of liposomes. These samples are used for Cyclosporin A treatment (inhalation) of lung tumors. I will also be using negative staining. Suggestions please!"
Thanks for any help with either of these questions -- Gerald Harrison
Can someone please enlighten me as to the difference in theory and practice between formalin and formaldehyde (presumably made up as a buffered solution) for tissue fixation. Dave
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut U-125 Storrs, CT 06269 Knecht-at-uconnvm.uconn.edu
Hi All, I recently got a software package for pc that is called "PAX IT". I haven't fully investigated all it's capabilities, but it seems like a very user friendly and well organized software. I am not sure about the MAc/Pc transfer, but you could ask them Midwest Information systems - tel. 847 455 0450.
cheers
Lucio
Dr.Lucio Mule'Stagno MEMC Electronic Materials Inc University of Missouri -St.Louis Silicon Materials Research Group Physics Dept., 501 Pearl Dr., 8001 Natural Bridge rd., St.Peters, St.Louis MO 63376 MO 63121 tel: 314 279 5338 314 516 5931/3 fax: 314 279 5363 314 516 6152 email: lmulestagno-at-memc.com luciom-at-newton.umsl.edu
On Wed, 15 Jan 1997, Ed J. Basgall wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } ----- Begin Included Message ----- } } We are looking for a powerful software to catalog, store and reteive } images. We prefer something that is compatible with both PC and the MAC. } Any suggestions? } } } __________________________________________ } Rex A. Hess, Ph.D., Associate Professor, } Director, Center for Microscopy and Imaging } University of Illinois, Veterinary Biosciences, 2001 S. Lincoln, Urbana, } IL 61802-6199 } 217-333-8933; FAX 217-244-1652; email: r-hess-at-uiuc.edu } homepage-- http://www.cvm.uiuc.edu/HomePages/rhess/hesspage.html } } ----- End Included Message ----- } } } Rex, } } I use Aldus Fetch as a cataloging program for my Power PC. The images can } be worked on with Photoshop on either PC or Mac. Image transfer } is usually accomplished by ftp from one computer to another over a network. } Others whom I know use it also like it. } } Ed Basgall, PhD } Res Assoc } Dept. of Chemistry } Surface Analysis Group } Penn State Univ. } 181 Materials Res. Inst. Bldg } University Park, PA 16802 }
POST-DOCTORAL RESEARCH ASSOCIATE Lehigh University
A post doctoral research associate position is available in the Department of Materials Science and Engineering at Lehigh University. The appointment is initially for one year, renewable for a second year. It involves an analytical electron microscopy study of boundary segregation in commercial aluminum alloys. A PhD in materials science and engineering with strong AEM background (X-ray and EELS) is essential: VG experience is highly desirable.
Please send resume and names and addresses of three referees to:
Dr. David B. Williams Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem, PA 18015
Lehigh University is committed to recruiting, retaining, and tenuring women and minorities.
Sharon L. Coe Department of Materials Science & Engineering 5 East Packer Avenue Lehigh University Bethlehem, PA 18015 610/758-5133 e-mail: slc6-at-lehigh.edu
I could use some help on identifying an ultramicrotome recently acquired from our life sciences department. Unfortunately, the previous owners did not have the instruction book for the instrument or any clue on how to use it. It was made by Cambridge and I believe the model is a Huxley ultramicrotome. I do have the serial number (sort of) written on the side of the instrument. Any help with locating the manufacturers or an instruction book would be greatly appreciated.
Any references on techniques for using the microtome with ultra-hard materials and composites would also be appreciated. Am I going to h-e-double-hockey-sticks for using a life sciences microtome in materials science?
Thanks,
Brian Gorman Graduate Research Assistant University of Missouri - Rolla Dept. of Ceramic Engineering 225 McNutt Hall Rolla, MO 65409
Tamara, We have been able to stain agarose (as well as some plant tissue) very well with fast green. Best results are to stain when the tissue is in 100% etoh. You can make a very concentrated solution of fast green (around 7% I think) and add a few ul per ml to the vial containing samples. This staining survives subsequent infiltration into methacrlate resins very well. Trouble is, if you are going through acetone, fast green is really not to solulble in acetone. I have not found a very good stain to use with acetone dehydrations, although Alizarin red isn't bad. Hope this helps, Tobias
Tamara, We have been able to stain agarose (as well as some plant tissue) very well with fast green. Best results are to stain when the tissue is in 100% etoh. You can make a very concentrated solution of fast green (around 7% I think) and add a few ul per ml to the vial containing samples. This staining survives subsequent infiltration into methacrlate resins very well. Trouble is, if you are going through acetone, fast green is really not to solulble in acetone. I have not found a very good stain to use with acetone dehydrations, although Alizarin red isn't bad. Hope this helps, Tobias
"Can someone please enlighten me as to the difference in theory and practice between formalin and formaldehyde (presumably made up as a buffered solution) for tissue fixation. Dave"
formalin is made by bubbling formaldehyde gas thru water until saturation (37% w/w or 40% w/v). Many (all?) commercial formalins contain 6-15% methanol to prevent polymer formation. Even with the methanol, polymers form over time. The different polymers presumably mean "different" fixation reactions are occuring. The formation of formic acid over time can cause the presence of "formalin pigment" due to reaction with hematin in blood rich tissues. Electron microscopists never (or virtually never) use formalin and use instead freshly depolymerized paraformaldehyde (heat granules to 60 C but not hotter, with stirring, then add a drop or two of 1 N NaOH - if you need a detailed protocol, e-mail me and I will send you one). One of the most common questions I get asked is whether it will make any difference in someone's LM immunocytochemical study to use freshly depolymerized formaldehyde as opposed to formalin. I am sure in many cases it will not make a difference but that there are undoubtedly examples where it does make a difference. I always make my formaldehyde fresh the day I use it.
Tom
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
Formaldehyde is a gas that is bubbled through water to give a 37-40% solution (saturated) sold as "formaldehyde". One part of this to nine parts water gives "formalin" or "10% formalin" (which is really 3.7 to 4.0% formaldehyde). When one makes a paraformaldehyde fixative it is often 4g paraformaldehyde per 100 ml of buffer.
Geoff (mcauliff-at-umdnj.edu) -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (908)-235-4583; fax -4029 e-mail: mcauliff-at-.umdnj.edu ***************************************************************
I seem to recall using dilute eosin, slightly acidified to keep it from leeching out, to stain agar blocks prior to embedding. Don't know what it might do to antigens, though.
Geoff (mcauliff-at-umdnj.edu) -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (908)-235-4583; fax -4029 e-mail: mcauliff-at-.umdnj.edu ***************************************************************
I know that this isn't specifically Microscopy but there may still be some strong interest.
Please send any questions or replies directly to john Vankat not myself.
John Vankat wrote: } } Assistant Professor, Botany } Miami University-Middletown } } The Department of Botany at Miami University seeks applicants for a } tenure-track position on the Middletown campus beginning August 1997. } Duties include teaching (occasionally at off-campus facilities) } introductory undergraduate lecture and laboratory courses in Botany } and participating in interdepartmental courses in biology. Other } responsibilities include service appropriate to the Regional Campus } mission and development of a scholarly program that involves } undergraduates. Position requires Ph.D. in Botany or closely related } discipline and experience in and commitment to undergraduate teaching } and advising. Send letter of application, one page statement of } teaching philosophy, description of teaching experience, curriculum } vitae, and three letters of recommendation to Dr. John L. Vankat, } Search Committee Chair, Department of Botany, Miami University, } Oxford, OH 45056. Review of applications begins February 3, 1997. } Phone: (513) 529-4200; Fax: (513) 529-4243; e-mail: } JLVANKAT-at-MIAMIU.MUOHIO.EDU } Miami University does not discriminate on the basis of sex, race, } color, religion, national origin, disability, age or sexual } orientation in its employment policies.
This message has also been posted to the confocal listserver.
Dear fellow microscopists,
Can someone enlighten me on the potential pitfalls of RI mismatch between coverslip and immersion oil. I am working with someone who must grow his cells on Aclar coverslips (RI = 1.435). We are mounting the specimens in Vectashield (RI = 1.4577) and are using Zeiss oil (RI = 1.515). We are trying to do confocal reconstructions of fluorescently tagged (FL and RH) mineral aggregates in these cultures (approx. 10-20 um} thick) to visualize their substructure. To do this we are using 100X oil immersion lens at Zoom 4 on a Leica TCS-NT confocal. What sorts of aberrations could I expect in the reconstructed images?? Should I use an oil with a lower RI? A related question: Most of the fluorescent specimens I work with have glass coverslips and are imaged with oil immersion objective lenses (consistent RI), but are mounted in Vectashield (Lower RI) or similar anti-photobleach medium. What problems does this pose for confocal imaging??
} Can someone please enlighten me as to the difference in theory and practice } between formalin and formaldehyde (presumably made up as a buffered } solution) for tissue fixation. Dave } Formaldehyde is a gas which, when dissolved to a final concentration of 40% in water, is termed formalin. Typically, commercially prepared formalin contains "stabilizers" such as methanol or calcium carbonate (chalk) which slow down the polymerization of the aldehyde groups. Formalin at 4-10% aqueous (buffered or unbuffered) may be suitable for preserving bulk specimens (chunks of liver, whole brains, etc) and may be suitable for preserving tissues for histological studies by light microscopy. For highest quality (ultrastructural studies, for example), it is best to prepare the formaldehyde by depolymerizing paraformaldehyde using a combination of heat and aldehyde (such as KOH). I can provide details, if you need this. Usually, one prepares an 8-10% aqueous solution of formaldehyde and then mixes this with a double strength buffer (phosphate buffer at pH 7.4 for mammalian tissues, for example) to obtain a buffered formaldehyde solution. Many buffers may be used and I can forward more info if you need it.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Southern Illinois University Carbondale, IL 62901-4402 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 ####################################################################
Due to policy and politics, our EM facility does not teach biological TEM "from scratch" (although we do teach SEM from scratch). We require users of most of our instruments to have had a course or prior experience. Right now there is no other place to learn TEM in our island state, and there is some pressure on us to teach several people. I am completely willing and able. However, in order to deal with the situation, we need to be able to 1) estimate about how much it would cost to train someone (or, more likely, a group of 4) to try to cover our actual costs and/or 2) where could these people go to take a short/intensive course (say, 3-5 weeks) and how much would that cost? There have been discussions here before about mixing new EM students with hard-core research interests. Up until now I've managed to keep it from being a problem, but now I am soliciting advice!
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangelo Text coming soon! **************************************************************************** * 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* ****************************************************************************
THis is a bit of an old saw but has anyone done a comparison of the currently available immersion oils wrt autofluorescence. . . . Simon ========================================= We supply the full range of Cargille immersion oils. Cargille know very well the properties of their oils and different grades are available for different purposes. Sufficient details for most people can be found in our on-line catalogue on page I1. The most commonly used oil is type B, which has "ideal" optical properties at average viscosity "low" auto-fluorescence. Type DF has "ideal" optical properties, "very low" auto-fluorescence but is a little more expensive. Type FF has "zero" auto-fluorescence with good (imperfect) optical properties at the same price as the DF. Labs using fluorescent and normal light microscopes and do not have several litre requirements of immersion oils annually may be best advised to use the DF type only. If they, however, require zero auto-fluorescence than it would be best to mark the bottles clearly and keep separate supplies.
As stated, we have a (small) interest in immersion oils. Jim Darley
Probing & Structure (Microscopy Supplies & Accessories) PO Box 111, Thuringowa QLD 4817 Australia
Phone +61 77 740 370 Fax: +61 77 892 313 A great microscopy site: http://www.ultra.net.au/~pns/
I am presuming you have some sort of tissue in the agar blocks. I have been using agar to encapsulate the tissue ( in most cases suspended cells). If you stain the agar with 1% eosin B (prepared in 100% alcohol) for about 5 to 10 mints and then rinse in 100% alcohol and then start the infiltration with LRWhite. I have had no problems so far with tissue loosing the antiginicity. If you need more details contact me.
Fellow microscopists, just an addition to the image archiving discussion: I use a similar program Graphic Workshop from Alchemy Mindworks, Beeton ON Canada LOG 1AD www.mindworkshop.com I had previously looked at the demo. version of Thumbsplus, and found it very similar, having purchased one, found no reason to migrate. Does anyone have a comment on relative capabilities? I am not connected with, or have any interest in either company. greetings to all from 37 oC (100 oF) Rio de Janeiro. Prof.Walter A.Mannheimer Metallurgy and Materials Engineering Federal University of Rio de Janeiro POBox 68505 21945 Rio de Janeiro, Brazil Voice +5521 280-7443 (Dept.office) +5521 590-0579 (direct) Fax +5521 290-6626 Email: wamann-at-metalmat.ufrj.br
} Can someone please enlighten me as to the difference in theory and practice } between formalin and formaldehyde ... } A saturated aqueous solution of formaldehyde contains (nominally) 37% formaldehyde. Such a solution is referred to as a 100% formalin solution. (A 10% formalin solution would contain ~3.7% formaldehyde.)
The presence or absence of other components (such as buffers) is irrelevant, other than affecting the final concentration of formaldehyde. } Cheers.
Don ______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 The College of New Jersey * fax: (609) 771-2674 Trenton, NJ 08650-4700
(* formerly Trenton State College; please note our new name)
We use Osmium to stain culture cells in agar, before we embed, then you can cut out the most concentrated area of cells to put in plastic.
Cheri Owen Detroit Neurotrauma Institute Detroit, Mi (313)577-4648
On Thu, 16 Jan 1997, Tamara Howard wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Probably a no-brainer :)...does anyone have any suggestions as to how I can } stain agar blocks for embedding in LR White? So I can find them again in } the final block? So far I've only tried Toluidine Blue - didn't work. I } have some Coomassie-agar in the works right now, but it is clearing, too. } Multiple suggestions are MORE than encouraged...because I'll ultimately } need something that will stain the agar without messing up my antigens. } Sigh. } Thanks! } Tamara } CSHL, NY } } }
I needed to stain agar several years ago. Rather than stain it, I found it better to incorporate a colored microsuspension into it. Blue dextran works fine...its used to visualize the solvent front in column chromatography and gel filtration. Its a high mw polysacharride, much like agar and is virtually innocuous. Sigma carries it.
-=W.L. Steffens=- Department of Veterinary Pathology College of Veterinary Medicine University of Georgia http://www.vet.uga.edu/wls/steffens.html
} } } A saturated aqueous solution of formaldehyde contains (nominally) 37% } formaldehyde. Such a solution is referred to as a 100% formalin solution. } (A 10% formalin solution would contain ~3.7% formaldehyde.) } } The presence or absence of other components (such as buffers) is } irrelevant, other than affecting the final concentration of formaldehyde. } }
I am afraid I strongly disagree with this statement. Some buffers can precipitate Ca2+ ions (e.g., PO4). The lack of Ca2+ in the fixative is well known to affect preservation. Some buffers with free amine groups can interact with the fixative. The presence of alcohol or other preservatives in commercial formalins could be expected to cause differences in immunoreactivity in some instances. I have seen some commercial formalin stocks that contained fluorescent impurities (tho this may have been contamination by users).
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
I have also used Graphics Workshop. I like to use it more for "manipulating" images. In that respects, use it as competition for commercially available graphics packages like Corel. I use Thumbs Plus primary for image databases (thumbnail subdirectories and contact sheets), archiving and quick looks at my data. I find Thumbs Plus to be much faster and easier to use for those operations.
At 11:21 AM 1/17/97 EST3EDT, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I would like to further the discussion on plasma cleaning, the need to clean specimen holders, and cleaning Be holder components. In the past I have found it extremely beneficial to pre-clean the TEM specimen holder from the vacuum o-ring to the tip. Although no one admits to touching the specimen holder, it does occur. In fact, after a few minutes of plasma cleaning, fingerprints become quite apparent on the specimen holder. I've also seen o-ring grease wind up on the shoulder of the rod.
In addition, specimen holders are often times stored in less than ideal conditions and surface contamination becomes inevitable. A recommended solution is to store the specimen holder under vacuum (oil-free) when not in use.
Another cause of specimen holder contamination is from adhesives which adhere to the specimen holder's clamping mechanism. Plasma cleaning with the clamping mechanism open is quite effective in removing this contamination. With the plasma flow being multi-directional, even hard to access areas of the specimen holder are cleaned.
The Be situation raises a much larger issue when discussing plasma. All types of plasma are not equal. Depending on the plasma generation system, high energy (} 100 eV) ions can be created. At this level, ion impingement results in the sputtering of the specimen, specimen holder, plasma chamber walls, and electrodes, if they too are immersed in the plasma.
The critical need for applying plasma to TEM specimens is to produce a plasma of sufficiently low energy so that it is below the threshold required to break a molecular bond (approximately 35 volts). One acceptable means of generating low energy ions is with a high frequency, inductively coupled plasma, whereby the electrodes are located external to the plasma chamber. As long as the ion energy is sufficiently low, plasma cleaning can occur without the risk of sputtering Be.
We have conducted measurements on the ion energies in our plasma cleaner and found them to be, under given conditions, in the 12-15 eV range, well below the sputtering threshold. In this energy range, a chemical reduction of the carbonaceous material occurs without altering the material's structure.
I hope that this information is helpful. Do not hesitate to contact me directly with any specific questions.
Kind regards,
Paul E. Fischione
E.A. Fischione is the manufacturer of the Model 1400 Plasma Cleaner and Vacuum Storage Containers.
It sounds like you've come across an old gravity-powered Huxley microtome. The specimen arm was lifted (and advanced) by a lever. When released, gravity pulled the arm down its cutting stroke, the speed of which was controlled by a variable oil-filled dashpot. Friction on the block-face had to be kept at a minimum so that it would not overcome the falling arm. You were limited to a very small block face and very thin sections. I suspect that it might have trouble with hard materials. If its all there, it should work, because it only had about 2 moving parts.
-=W.L. Steffens=- Department of Veterinary Pathology College of Veterinary Medicine University of Georgia http://www.vet.uga.edu/wls/steffens.html
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Gerald,
Years ago I processed synaptosomes for thin section EM by "packaging" them in tiny agar tubes; I would try to do the same with your liposome preps.
Start with a 5% agar solution at ~60 degrees; thin down if tube walls are too thick. Dip a glass micropipet (type used for hematocrits) into the agar, cool over ice and slide the agar off the pipet with your fingers onto a cool wax sheet (sitting on ice). Cut the tube into bits ~0.5 cm long. Put a drop of your material onto the wax, pick up the tube and fill by touching to the drop. Seal with just-molten agar; trim off the "dumbbell" ends with a razor and process the filled tubes as you would a piece of tissue. Issues are the finding the right concentration of agar to yield some tension in the walls of the tube but yet thin enough for good permeability. Temperature may be tricky with liposomes.
WANTED: MONOCULAR HEAD or ADAPTER (that will fit INSIDE the binocular eye-piece tube) for CARL ZEISS GFL microscope to enable me to set up for microphotography. Adapter must be able to take a standard T-mount for a 35mm camera. Hopefully, you have an item of this sort gathering dust and can let it go at a very low price (plus shipping). Thank you. E-mail: bobcat54-at-aol.com
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Fellow Microscopists, I'm looking for a full time technician position in a EM lab. I possess an appropiate educational background that match most of requesting in regard to the employment at EM laboratory. I have an EM Technician Certificate (1 yr. postgraduate program at Seneca College, Toronto, Canada)+ M.Eng. degree from Technical Academy of Agriculture, Faculty of Zootechny, Olsztyn,Poland. In addition, the EM program from Seneca College, Toronto gave many students untill closing hands on experience and theoretical knowledge in operation of TEM-SEM including all related techniques of biological specimen preparation essential to subsequent EM observation. My career objective is to become one of you with wide variety of preparation techniques in a biological field of EM. $ Any assistance you might be able to offer will be greatly appreciated or you need the further information about me just replay for this message. FOR ALL OF YOU I WISH A HAPPY & PROSPEROUS 1997! Andrew Kuczynski E-mail: 102137,1277-at-compuserve.com *************************************************************************** "While my name maybe is foreign to you, our language is the same and our backgrounds are similar." ***************************************************************************
I'm about to open up for the first time the diff pump in the carbon coater which I inherited as a going concern some 8 years ago. I suspect that the fluid in it is silicone, possibly DC 704, the one thing I do know is that it'll be pretty dirty. Does anyone know an infallible way to test very used diff pump fluid to ascertain whether it is a silicone or not?
cheers
Ritchie
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
I am trying to find a way to measure and compare the relative intensity or brightness (?) of images (photomicrographs) of fluorescent labeled cells.
I have NIH Image as well as Photoshop and Excel. Would it be possible to make these types of measurements using these programs ? Is there another program you would recommend ?
What I am doing more specifically, is labeling cells at different micromolar concentrations of the fluorescent marker PKH26, then putting the cells into tissue culture flasks. I will then take slide photographs os the the flasks under magnification and fluorescent illumination at different points in time to assess the falloff in flurescence. I have the capability of making scans of the slides.
Thank you very much for your time,
David Fox, MD
************************** David Fox, MD Fellow in Vascular Surgery Loyola University Medical Center
For over 15 years now I have stored stages in slightly over pressurized dry Nitrogen environment using the N2 gas which vents off of LN2 tanks here at ANL. The N2 is directed into a simple plexiglass storage boxes with simple seals on the doors. These can be purchased from all the standard Microscopy Supply Houses. It is my experience that vacuum storage is rarely necessary and I've never seen documented evidence to show vacuum storage of TEM stages reduces contamination.
Having said this I do store a few stages in mild vacuum, but for very different reasons. The exception here, are some old Gatan LN2 & LHe stages we have here at ANL, which use microscope vacuum to achieve insulation. These (very) old models evacuated an outer insulating chamber of the cooling dewar by pumping on it via the microscope column using a small hole judiciously placed on the "column side of the o-ring seal". Leaving these stages in air always increases the pump down time when the stage is inserted into the microscope, presumably due to water vapor collecting in the chamber. Gatan has since redesigned their stages to remove this problem, however, I still use these stages as they continue to work. In this case the stages are stored in a simple chamber pumped down to ~ 10 mT using any RP. They are leaked back to ATM using N2 to make sure H2O vapor does not collect in the insulating dewar. I have never seen evidence of contamination of specimens which can be attributed to the stage being stored in a standard RP system, when the RP is properly operated. Of course, if you screw up and backstream oil into the system then all bets are off, however, that is a simple thing to avoid by good laboratory practice.
Our old Edwards 12E6 from 1966 was overhauled, upgraded etc. in 1981 and charged with 100 ml Edwards Silicone 704/F4. It reaches 1x10-4 torr during a normal day and 1x10-5 torr when cryo activities are going on. It is used for cleaning jobs and carbon filming. Not great maybe but the figures have held constant since the beginning. A while ago I intended opening it up again (it used to be an annual event pre-1976 with my predecessor) but on his retirement, time became more precious.
I intended opening it up a few years ago because a young technician came and said 'Keith, I think something might be wrong with the coating unit because there is smoke coming out the back' !! It turned out that the system was pumping in hi-vac mode and she had opened the air inlet! It was oil mist and I guessedthat the oil charge had by then disappeared. But no, it still runs. You've pricked my conscience! But those old units run and run!
With best wishes
Keith Ryan Plymouth Marine Laboratory, Citadel Hill, Plymouth, Devon PL1 2PB, England
Hi All For teaching purposes to show simple diffraction patterns and tilting thereoff, as well as to demonstrate twinning we chose the old route of preparing thin (20nm) gold crystal by by evaporating firstly Ag then Au on air cleaved NaCl at 400 to 450 degrees C (10-5 torr). We experience a few problems to get reproducible crystals: 1. Sometimes the Ag surface turn whitish in stead of silver. 2. The Ag does not completely dissolve in nitric acid (various concentrations tried...).
Can somebody help or refer me to literature on the subject or maybe suggest other routes for simple preparation of such demonstrating foils?
Thanks in advance
Sara Prins Surface and Structure Analytical Services Division for Material Science and Technology CSIR PO Box 395 Pretoria 0001, South Africa +27+12+8413974 sprins-at-csir.co.za
} I recently received the following request from a non-microscopist } colleague: } } } Do you have any current information about formaldehyde & cancer? } } We're looking for publications/documentation
Here're the most recent refs from a lit. search I did a while ago:
Authors Anonymous. Title Formaldehyde. [Review] Source IARC Monographs on the Evaluation of Carcinogenic Risks to Humans. 62:217-375, 1995.
Authors Hansen J. Olsen JH. Title Formaldehyde and cancer morbidity among male employees in Denmark. Source Cancer Causes & Control. 6(4):354-60, 1995 Jul.
Authors McLaughlin JK. Title Formaldehyde and cancer: a critical review. [Review] Source International Archives of Occupational & Environmental Health. 66(5):295-301, 1994.
Authors Sterling TD. Weinkam JJ. Title Mortality from respiratory cancers (including lung cancer) among workers employed in formaldehyde industries [see comments]. Source American Journal of Industrial Medicine. 25(4):593-602; discussion 603-6, 1994 Apr. Comment in: Am J Ind Med 1995 Feb;27(2):301-5
Authors Marsh GM. Stone RA. Esmen NA. Henderson VL. Title Mortality patterns among chemical plant workers exposed to formaldehyde and other substances [see comments]. Source Journal of the National Cancer Institute. 86(5):384-6, 1994 Mar 2. Comment in: J Natl Cancer Inst 1994 Oct 19;86(20):1556-8
Authors Gardner MJ. Pannett B. Winter PD. Cruddas AM. Title A cohort study of workers exposed to formaldehyde in the British chemical industry: an update. Source British Journal of Industrial Medicine. 50(9):827-34, 1993 Sep.
Authors Partanen T. Title Formaldehyde exposure and respiratory cancer--a meta-analysis of the epidemiologic evidence. Source Scandinavian Journal of Work, Environment & Health. 19(1):8-15, 1993 Feb.
Authors Luce D. Gerin M. Leclerc A. Morcet JF. Brugere J. Goldberg M. Title Sinonasal cancer and occupational exposure to formaldehyde and other substances. Source International Journal of Cancer. 53(2):224-31, 1993 Jan 21.
Authors Marsh GM. Stone RA. Henderson VL. Title Lung cancer mortality among industrial workers exposed to formaldehyde: a Poisson regression analysis of the National Cancer Institute Study. Source American Industrial Hygiene Association Journal. 53(11):681-91, 1992 Nov.
Authors Marsh GM. Stone RA. Henderson VL. Title A reanalysis of the National Cancer Institute study on lung cancer mortality among industrial workers exposed to formaldehyde. Source Journal of Occupational Medicine. 34(1):42-4, 1992 Jan.
Authors Holmstrom M. Lund VJ. Title Malignant melanomas of the nasal cavity after occupational exposure to formaldehyde [see comments]. Source British Journal of Industrial Medicine. 48(1):9-11, 1991 Jan. Comment in: Br J Ind Med 1993 Aug;50(8):767-8
Authors Partanen T. Kauppinen T. Hernberg S. Nickels J. Luukkonen R. Hakulinen T. Pukkala E. Title Formaldehyde exposure and respiratory cancer among woodworkers--an update. Source Scandinavian Journal of Work, Environment & Health. 16(6):394-400, 1990 Dec.
Authors Blair A. Saracci R. Stewart PA. Hayes RB. Shy C. Title Epidemiologic evidence on the relationship between formaldehyde exposure and cancer. [Review] Source Scandinavian Journal of Work, Environment & Health. 16(6):381-93, 1990 Dec.
Authors Blair A. Stewart PA. Hoover RN. Title Mortality from lung cancer among workers employed in formaldehyde industries. Source American Journal of Industrial Medicine. 17(6):683-99, 1990.
Authors Boysen M. Zadig E. Digernes V. Abeler V. Reith A. Title Nasal mucosa in workers exposed to formaldehyde: a pilot study. Source British Journal of Industrial Medicine. 47(2):116-21, 1990 Feb.
I hope these have what your colleague is looking for.
Leon
-- Leon A. Metlay, M.D.,Associate Professor of Pathology and Laboratory Medicine University of Rochester Medical Center Phone: (716) 275-5691 P.O. Box 626 Fax: (716) 273-1027 Rochester, NY 14642 lmetlay-at-acu.pathology.rochester.edu http://www.urmc.rochester.edu/smd/pathres/URPLM.html "Most ass drivers are evil, most camel drivers are decent, most sailors are saintly, the best among physicians is going to Gehenna, and the best of butchers is a partner of Amalek" -R. Judah, in Mish. Kidd. 4:14
Considering the current, and potential future state of Apple computers, I wonder how many people out there would be interested in a PC compatible interface for a Gatan PEELS. I am not trying to sell one, would like one myself; the target is that if a number of other people are interested in might be possible to push harder various companies to market such an interface.
} Hi All } For teaching purposes to show simple diffraction patterns and tilting } thereoff, as well as to demonstrate twinning we chose the old route of } preparing thin (20nm) gold crystal by by evaporating firstly Ag then Au } on air cleaved NaCl at 400 to 450 degrees C (10-5 torr). } We experience a few problems to get reproducible crystals: } 1. Sometimes the Ag surface turn whitish in stead of silver. } 2. The Ag does not completely dissolve in nitric acid (various } concentrations tried...). } } Can somebody help or refer me to literature on the subject or maybe } suggest other routes for simple preparation of such demonstrating foils? } } Thanks in advance } } Sara Prins } Surface and Structure Analytical Services } Division for Material Science and Technology } CSIR } PO Box 395 } Pretoria } 0001, South Africa } +27+12+8413974 } sprins-at-csir.co.za
Can't help with the gold, but I'd recommend molydenum oxide for similar uses, and it's a lot easier to prepare. Slowly heat a molybdenum strip or length of wire in air until a white smoke comes off. Wave a grid through the smoke. That's it - the grid is covered in molydenum oxide crystals, of various sizes, some twinned. Nice, easy single crystal diffraction patterns and you can use the specimen to calibrate the rotation relationship between the image and diffraction pattern.
Before I am completely swamped by emails, I am not talking about a sophisticated and expensive system that controls the microscope as well as the PEELS (i.e. the EMiSPEC system) but something a little more modest that only controls the PEELS, and does not cost $50K ! /
I too would be interested in knowing a simple way to distinguish among the common types of diffusion pump oils. One way to tell if you have a silicone oil might be to smear a bit of it on a metal stub and run an EDX spectrum of it to see if it contains Si (Maybe it would be better to burn some in a platinum or tungsten boat and examine the ash for Si). However, there is undoubtedly someone out there who will know a simpler method.
It is true that diffusion pumps will often perform at an acceptable level, even after thay have been savagely mistreated. However,after a diffusion pump has been run for any significant period of time without proper cooling water,or after exposure to air at high pressures, or after failure of the mechanical backing pump, the oil in it is likely to have been depleted, oxidized, and/or thermally degraded enough so that the pump no longer performs optimally, and is highly likely to exhibit an abnormally high level of backstreaming. (See 'Vacuum Methods in Electron Microscopy', p 214-220 for a detailed discussion.) The silicone and perfluorinated polyphenyl ether oils are the most resistant to such degradation, the polyphenyl ether oils are pretty good, while the synthetic ester and hydrocarbon fluids are much less so (p. 180-188). In any event, it is best to service a pump as soon as possible after such a potentially damaging event has occurred, just to minimize the liklihood that the vacuum system will become seriously contaminated with degraded pump oil.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
I wish to offer a correction and apology for my sin of omission.
In some comments I made over this Listserver a few days on methods for cleaning TEM specimen holders ago I stated that, "The latest method method for these holders is Plasma Discharge Cleaning, and Southbay Technologies markets a device that is specially designed for this purpose". Insofar as I know this statement is correct (except that the name should be 'Southbay Technology'); however, I have been reminded that the method was developed by Nestor Zaluzec and patented by Argonne Nat'l. Lab., and that two other companies also market Plasma Cleaning devices: namely, SPI Supplies and E. A. Fischione. This was an oversight on my part, caused in part by the fact that I was a bit loose in the wording of my comments, but also because I apparently don't keep very up-to-date on the latest developments in the marketplace for EM supplies (with my luck, two or three other companies are probably also in the buisness by now, and so I'll be off again).
In any event, I have no commercial interest in this matter, and was not trying to offer information prejudicially favoring any particular company over any other. I've been involved in the field of electron microscopy for so long that I have friends in most companies associated with the field, and so my intention is to remain as unbiased as possible. I'll try to be more careful henceforth.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
1) We would like to contact Beverly Giammara for some help and information regarding her publications on flat molds for embedding tissue using the microwave. If any one has any information on how to contact her we would greatly appreciate it.
2) To any one else doing microwave embedding
We are a lab that deals with muscle and nerve samples. We would like to use our new microwave to embed the tissue in resin (Spurr's or LR White). We tried Beem capsules but were unable to orient the tissue properly for sectioning with our MT2-B. I have seen reference made to the use of flats molds in the microwave and wonder if any one has any experience or suggestions they could share with us.
Thanks in advance
Christine Brantner and Susan Danielson Muscle/Nerve Lab Froedtert Hospital Milwaukee WI
We are interested in which lazer printers people have had experience using for routine biological EM images. Our images are captured with a Kodak Megaplus 1024x1024. We would also use the printer for LM, anatomical line drawings, AR of gells etc. We are considering the Lexmark Optra R+ 1200 dpi. Thanks for your help.
Rick L. Vaughn EM Research Facility Dept. Cell Biology & Anatomy Univ. Neb. Med. Ctr.
Hi everybody, And specially people working on asbestos. I would like to know which method is used in the other countries (USA UK Canada Germany....) to determine the concentration of asbestos in atmosphere. By method I mean when a laboratory receive a filter from a suspected place do they use a TEM or SEM for counting fibers and how? Do they use electron diffraction or EDS to determine the nature of asbestos? Thank you very much for the answers. ========================================================== Jacky Larnould tel 33 (0)4 67 72 28 26 fax 33 (0)4 67 79 54 90 email larnould-at-mnet.fr
Does anyone have any experience with SEM of reconstituted rat tail collagen matrix? The collagen is laid down onto glass coverslips and then cells are plated on top of the matrix. I am interested in the collagen more than the cells, but can't get any definition of fibers. In some areas there is some definition, but the majority of the surface seems compacted with little or no definition. I have tried a variety of preparation conditions including different fixatives as well as cpd vs. freon for drying. The latter seemed to produce the best results, but there is still little definition of the matrix. I have sputtered for different times, but I still seem to have a problem of charging that eventually burns the sample no matter what kV I use. Does this sound like a preparation problem, an imaging problem, or both? Any help would be greatly appreciated. Thanks. Bill Swaim
Hi, I am looking for any publications which contain micrographs of plant cell necrosis. I am specifically interested in sclerenchyma cells, but any papers on necrosis would be helpful. Thank you. Debby LeBlanc
} } We are interested in which lazer printers people have had experience } using for routine biological EM images. Our images are captured with a } Kodak Megaplus 1024x1024. We would also use the printer for LM, } anatomical line drawings, AR of gells etc. We are considering the } Lexmark Optra R+ 1200 dpi. Thanks for your help. } } Rick L. Vaughn } EM Research Facility } Dept. Cell Biology & Anatomy } Univ. Neb. Med. Ctr.
We use the Lexmark Optra RX printer in our lab for routine image printing and we are very happy with it. We are a materials lab here but I think you would be happy with it for your biological images as well.
Good Luck,
Margaret E. Bisher
NEC Research Institute 4 Independence Way Princeton, NJ 08540. Tel.: (609) 951-2629 Fax: (609) 951-2496 e-mail: peggy-at-research.nj.nec.com
I have a JEOL 840 SEM that was purchased in 1986, and we are considering putting it up for sale. I would like some feed back on what this instrument might be worth or what you a potential buyer would offer if interested.
The scope has both SE and BSE detectors, plus a PGT System III X-Ray detector. The scope is currently in operation and has been under service contract with Jeol for the last 5 years., Overall it is in excellent condition.
Please respond directly to me and not this list either by e-mail, phone or fax.
Thank you.
Joe Goodhouse Confocal / E.M. Core Facility Dept. of Molecular Biology Princeton University
It would seem to me that the simplest way of determining if silicone oil is in the diffusion pump (as opposed to something else?) is to take an Infrared scan - silicone oil has very distinctive absorption bands in the "fingerprint" region of about 1450 - 700 wavenumbers. Examples can be found in just about any IR reference book. One could also monitor the quality of the oil by taking an initial IR and periodically checking subsequent spectra against it.
Regards,
-Bob ********************************** Bob Citron Chiron Vision 555 W. Arrow Hwy Claremont, CA 91711 USA PH: (909)399-1311 Email: Bob_Citron-at-cc.chiron.com **********************************
We have been using a red new fuschin/alkaline phosphatase reaction to tag a cytoplasmic protein. It transmits light maximally at 600 nm, and fluoresces with a rhodamine cube.
We are looking for similar dual-function chromophores to label cytoplasmic proteins and nucleotides.
Susan Danielson writes: } 1) We would like to contact Beverly Giammara: Her email address is } {bgiammara-at-magnum.mco.edu}
} 2) To any one else doing microwave embedding: } We are a lab that deals with muscle and nerve samples. We would like to } use our new microwave to embed the tissue in resin (Spurr's or LR White). } We tried Beem capsules but were unable to orient the tissue properly for } sectioning with our MT2-B. I have seen reference made to the use of } flats molds in the microwave and wonder if any one has any experience or } suggestions they could share with us.
RESPONSE- There are two approaches in the literature for microwave accelerated curing of resins: 1) Giammara's approach for flat embedding molds and 2) Giberson and Demaree's approach for Beam Capsules.
My suggestion for flat embedding molds is to start with 50% power for 15 minutes (100% will definately give you disappointing results). Also when using flat embedding molds, allow your blocks to cool for 15 minutes before removing them from their molds, and vent your microwave oven during curing.
I highly recommend using an appropriately sized water load for your oven during microwave curing in flat embedding molds (otherwise there is simply too much energy in a microwave oven and specimen damage from over heating will result). In addition, microwave curing in an uncalibrated microwave oven is very tricky and usually results in disappointing results (e.g., incomplete curing of blocks). Simple tools that you can make and a detailed description of how to use them to CALIBRATE YOUR MICROWAVE OVEN are published in the The Microwave Tool Book.
When curing resins in BEEM capsules, they are placed IN a water bath- temperature never exceeds 100 =B0C and curing times are ~ 75 minutes. See the reference by Giberson RT, Demaree RS, Jr: Microwave fixation: understanding the variables to achieve rapid reproducible results. Microsc Res Tech 32:246, 1995
Detailed information regarding curing times of various resins in a microwave device can be found in: 1. Giammara B. Microwave embedment for light and electron microscopy using Epoxy resins, LR White, and other polymers. Scanning 1993;15:82-87.
2. Login GR, Dvorak AM. The Microwave Toolbook. A Practical Guide for Microscopists (Beth Israel Hospital, Boston, 1994). The book contains a table of curing times for resins tested.
Please contact me if you have additional questions and I will respond directly to you.
Dr. Gary R. Login Dept. Pathology Beth Israel Deaconess Medical Center 330 Brookline Avenue Boston, MA 02215
Susan Danielson writes: } 1) We would like to contact Beverly Giammara: Her email address is } {bgiammara-at-magnum.mco.edu}
} 2) To any one else doing microwave embedding: } We are a lab that deals with muscle and nerve samples. We would like to } use our new microwave to embed the tissue in resin (Spurr's or LR White). } We tried Beem capsules but were unable to orient the tissue properly for } sectioning with our MT2-B. I have seen reference made to the use of } flats molds in the microwave and wonder if any one has any experience or } suggestions they could share with us.
RESPONSE- There are two approaches in the literature for microwave accelerated curing of resins: 1) Giammara's approach for flat embedding molds and 2) Giberson and Demaree's approach for Beam Capsules.
My suggestion for flat embedding molds is to start with 50% power for 15 minutes (100% will definately give you disappointing results). Also when using flat embedding molds, allow your blocks to cool for 15 minutes before removing them from their molds, and vent your microwave oven during curing.
I highly recommend using an appropriately sized water load for your oven during microwave curing in flat embedding molds (otherwise there is simply too much energy in a microwave oven and specimen damage from over heating will result). In addition, microwave curing in an uncalibrated microwave oven is very tricky and usually results in disappointing results (e.g., incomplete curing of blocks). Simple tools that you can make and a detailed description of how to use them to CALIBRATE YOUR MICROWAVE OVEN are published in the The Microwave Tool Book.
When curing resins in BEEM capsules, they are placed IN a water bath- temperature never exceeds 100 =B0C and curing times are ~ 75 minutes. See the reference by Giberson RT, Demaree RS, Jr: Microwave fixation: understanding the variables to achieve rapid reproducible results. Microsc Res Tech 32:246, 1995
Detailed information regarding curing times of various resins in a microwave device can be found in: 1. Giammara B. Microwave embedment for light and electron microscopy using Epoxy resins, LR White, and other polymers. Scanning 1993;15:82-87.
2. Login GR, Dvorak AM. The Microwave Toolbook. A Practical Guide for Microscopists (Beth Israel Hospital, Boston, 1994). The book contains a table of curing times for resins tested.
Please contact me if you have additional questions and I will respond directly to you.
Dr. Gary R. Login Dept. Pathology Beth Israel Deaconess Medical Center 330 Brookline Avenue Boston, MA 02215
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We have an Hitachi SEM that uses threaded stubs. On the suggestion of Kevin Cronyn (Hitachi Sales) I bought plastic hinged-lid boxes (about 4" x9") from one of the EM suppliers and cut pieces of plexiglass to fit in the bottom. I then drilled and threaded 32 holes in the plexiglass and ran short (~1/2") bolts up through the holes. The thread size is the same as for the Hitachi stubs so you just screw the stubs down on the bolts and set the whole unit in the plastic box. I put one longer (~1") screw in the middle to act as a handle for getting the plexiglass out of the box. I seem to recall that it came to about $20 in supplies for each box as well as two hours of drilling and tapping a bunch of little holes. I can send more details if you are interested. For short-term student use I give them petri plates with double-sided tape in the bottom. Stubs are placed on the tape and stick pretty well. You can get up to 10 or so stubs in one standard Petri dish.
YWIA
Bob
Robert R. Wise Plant Physiologist and Director, UWO Electron Microscope Facility Department of Biology University of Wisconsin Oshkosh Oshkosh, WI 54901 (414) 424-3404 tel (414) 424-1101 fax wise-at-uwosh.edu
Can anyone out there direct me towards a decent monte carlo electron- specimin interaction modeling program? I'm looking for electron path, BSE energy and directional information. Provided that the code is in either fortran or C++ (or basic, I guess), I will be able to make minor modifications if the program(s) are not precisely suited to these uses. thanks, Ben Simkin (simkin-at-egr.msu.edu)
Dear Christine and Susan, and fellow microscopists,
A good introduction to Beverly Giammara's work with microwave embedding can be found in Chapter 23 of The Microwave Cookbook for Microscopists, by Kok and Boon, entitled "Microwave Exposure and Epoxy-resin Embedding for EM." This book is available through Energy Beam Sciences.
We also have a bibliography of papers relating to this subject at our World Wide Web site (http://www.ebsciences.com).
Using the method developed by Giammara, Spurr or LR White resin polymerization can be done in about 30 minutes in a laboratory microwave.
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
} What is the best way to store SEM samples (those already on stubs and } sputter coated)? } Best way we have found is to use storage boxes provided by SPI, EMS or Pella (clear plastic boxes) and place in either a glass desiccator or zip-lock bag with desiccant. The boxes offer the advantages: numerically labeled for specimen ID, hold the specimens tightly so that they will not spill out if inverted or tipped.
A cheaper alternative is to take some 1/2" plexiglass and drill a series of holes that will allow the stubs to fit. A dab of sticky tab will adhere the stub to the plexiglass. If one attaches small legs, the plexiglass panels may be stacked on top of each other. This may then be desiccated.
Cheapest yet, someone (EMS or Pella or SPI) makes some paper boxes (like "pill boxes" of olden days) that you can write on.
Good luck.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Southern Illinois University Carbondale, IL 62901-4402 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 ####################################################################
Dear Ben, The program Electron Flight Simulator will do all that and some other neat things besides. You will find references in any Microscopy Today or the Exhibitors Bulletin of the last MSA meeting and there is a Web site. It costs about $480 and runs on a PC. Other than that, David Joy is the writer of most of those programs, and should be able to help you with a free version. You wrote: } Can anyone out there direct me towards a decent monte carlo electron- } specimin interaction modeling program? I'm looking for electron path, } BSE energy and directional information. Provided that the code is in either } fortran or C++ (or basic, I guess), I will be able to make minor modifications } if the program(s) are not precisely suited to these uses. } thanks, } Ben Simkin (simkin-at-egr.msu.edu) } Regards, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
Here at the museum our SEM samples are frequently from registered specimens from the collections and are often the type specimens of new species. Our stubs are, therefore, stored "in perpetuity" like the rest of the collections. Years ago I asked some questions regarding conditions for permanent storage but there wasn't much info forthcoming apart from the usual methods. For what it's worth, here are some of my observations on stubs 15-20 years old.
The oldest stubs in our museum have been stored in large (~300mm diam.) glass petri dishes, stuck down on double-sided sticky tape. These petri dishes are kept in stacks of three in glass dessicators (with silica gel) which are sealed with petroleum jelly. Most of them are still good for the SEM - the bad ones are attributable to poor preparation (and subsequent degradation) rather than storage conditions. Others have used disposable plastic 110mm dishes, however, this is not so space-efficient.
I guess that low humidity and constant temperature are the important factors. I've thought (comments please) that it might be worth replacing the air with an inert gas as well.
We then looked at perspex cabinets with shelves. None were to our liking (usually too few shelves and too expensive) so we designed our own for a person who wanted to produce them for his supply shop.
Originally we had planned for a stackable perspex cabinet (340mm wide, 280mm deep, 260mm high) with O-ring in the door, full-length side hinge, roller clips to provide pressure on the O-ring, gas exchange taps (pump inert gas in through one and air out the other) and 10 shelves which held 1,760 stubs. Unfortunately, supply of parts and costs for materials and labour, on what was only ever going to be a small run, meant considerable changes and the result can only be called a very efficient dust cabinet (still stores 1,760 stubs!). With monthly changes of silica gel it works as a dessicator.
Any supply houses interested in bringing this design to fruition?
Geoff Avern Microscopy Laboratories Australian Museum Sydney, Australia
P.S. Paula, please say hello to Carole Hickman (Palaeontology) if you see her.
I am presently reassessing the anti-corrosion/anti-microbial growth additive to use in our two EM cooling systems. The questions that I am having difficulty in finding answers to are;
1)What are the properties of water that cause corrosion in the electron microscope? Is it the pH, water contents or both?
2)What is the best pH for EM cooling systems?
3)Has anybody used the Coalite Chemicals product 'Phylatol' in their EM cooling systems?
Thanks in advance.
Allan Mitchell
Please send replies to: allan.mitchell-at-stonebow.otago.ac.nz
Does anyone know of any critiques comparing/contrasting the various commercial & shareware packages for microscopy such as Metamorph, Metafluor, NIH Image, Axon Image Workshop, Global Image, Image Tools (Un. of Texas), etc.?
Is this the appropriate forum for such a comparision, or does it exist elsewhere? Thanks, Sandy Simon
Sanford M. Simon Laboratory of Cellular Biophysics Box 304 Rockefeller University 1230 York Avenue New York, N.Y. 10021 (212) 327-8130 (voice) (212) 327-8022 (fax) simon-at-rockvax.rockefeller.edu (e-mail)
"Well I ain't often right, but I've never been wrong It seldom turns out the way it does in the song, Once in awhile you can get shown the light in the strangest of places if you look at it right..."
} What is the best way to store SEM samples (those already on stubs and } sputter coated)?
My first thought was "In peanut butter jars with dessicant, of course!" because that's what I just told my new SEM class and what I tell everyone else. Our main concern here in Hawaii is humidity, and I insist that all samples be held over dessicant for some time before going into our field emission SEM. Most of us put our pin-style stubs in commercially available boxes and find that, once sputter coated, they will then store *indefinitely* over dissicant. The identification and procurement of suitable jars is a serious subject here, especially now that many brands of peanut butter are now available only in plastic jars with a thin, styrofoam-like seal, driving us to other snacks that come in jars with the requisite rubber ring in the lid. Jellies and pickles and other fluid items frequently come in jars with wide mouths, allowing room for insertion of fingers to retrieve sample storage boxes. Mayonnaise jars, alas, do not have the rubber ring in the lid. Canning jars are popular among our customers with active grants to pay for them. Be warned, however, against using jars which may have strong residual scents; kim chee jars are a no-no. Avoid, too, jars which have contained spaghetti sauce or other tomato-based products as some strange mungy stuff likes to grow in them even after being subjected to multiple runs in the dishwasher. I have not attempted to autoclave them.
Tupperware brand storage boxes work well for a fair period of time; other brands do not seal well enough to keep indicator dessicant from indicating. My Tupperware lady thinks I'm nuts. She may be right.
Jars of all shapes and sizes full of stub boxes pile up in our facility until I run down their long-lost owners or shove them in their campus mail boxes. I can't help you with that problem!
If the question is, rather, how does one keep the stubs upright and undamaged if they are the type that does not fit snugly in comercially available storage boxes, or one can't afford said boxes, I am of less help. I like to encourage my customers and students to be creative (I have an interesting collection of boxes designed to hold glass knives, for example). I have not yet tried to see if 1/8" pin-type stubs fit in the holes of pipettor tip boxes. Hitachi screw-on stubs are such a pain that I made an adaptor to hold pin-type stubs before I took delivery of the 'scope. I remember that you have an ISI DS-130, but I don't remember the stub type.
Last week I looked at some stubs of unknown origin that had been knocking around in a petri dish in a desk drawer for at *least* 12 years. I put them over silica gel for a day and popped them into the 'scope without further coating and found some wonderful cultured cells that looked just great!
I'd rather be in the lab than home with this cold. (It's raining in Hawaii.)
Tina
MicroAngelo soon admitting gender and changing to MicroAngela http://www.pbrc.hawaii.edu/bemf/microangelo **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
What is the best way to store SEM samples (those already on stubs and sputter coated)?
We have chosen the expensive way by storing the samples in a vacuum cabinet. The vacuum is not very good (rough pumping) but sufficient to prevent damages of the sputtered layer, water vapor uptake and/or oxidation of the samples. As our (Philips) stubs have a pin underneath, we have drilled holes in the cabinet's shelves. This way the samples are fairly secured when you move the shelves. This storage is meant for samples we might want/need to put back in the microscope. Once they are no longer current, we have a large drawer with a rubber foam (neoprene) layer in which we've drilled holes too so that the pin stubs will fit. It works very well and if you reference your shelves and drawers, you might even be able to actually find old samples back: -).
Have a nice day
J.-M. Boichat e-mail: jean-marc.boechat-at-chma.mhs.ciba.com EM LABS Ciba Research Center phone:++41264356979 fax:++41264356907 P.O. Box 64 1723 Marly 1 Switzerland Disclaimer: Nobody in this company ever mind what I say why would they start now!
Message-Id: {1.5.4.32.19970122140553.006bd218-at-biotech.ufl.edu} X-Sender: gwe-at-biotech.ufl.edu X-Mailer: Windows Eudora Light Version 1.5.4 (32) Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
For short term storage we have used several of the methods already described, depending on the state of our budget at the time. For extended storage of samples that you really don't expect to ever look at again but someone insists that you archive, we have taken to using the commercial storage box placed in a seal-a-meal bag with some charged silica gel. The bag is evacuated and sealed for storage on a high shelf in the lab. The indicator in the silica gel shows that it is still dry after three years. ******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
What is the best way to store SEM samples (those already on stubs and sputter coated)?
We have chosen the expensive way by storing the samples in a vacuum cabinet.
The vacuum is not very good (rough pumping) but sufficient to prevent damages of the sputtered layer, water vapor uptake and/or oxidation of the samples.
As our (Philips) stubs have a pin underneath, we have drilled holes in the cabinet's shelves. This way the samples are fairly secured when you move the shelves. This storage is meant for samples we might want/need to put back in the microscope. Once they are no longer current, we have a large drawer with a rubber foam (neoprene) layer in which we've drilled holes too so that the pin stubs will fit. It works very well and if you reference your shelves and drawers, you might even be able to actually find old samples back: -).
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
To: Microscopy ListSer {Microscopy-at-MSA.Microscopy.Com}
Yes, we're interested in SW for image capture process and print. Please include image capture cards.
Right now, we're hung up with micro-channel bus computers and old IBM Video Capture Adapter cards. The greatly restricts the choices. We use Perfect Image/2 to capture, but we have to save the images and open them with Paint Shop Pro in order to anotate and print them. Our Lexmark Optra 4049 gives us nice 1200 dpi prints on plain paper. This system is complicated to use, but works. We may try to get ISA bus machines and new capture cards if it looks worthwhile.
Please tell us where there might be comparisons of cards and sw packages for SEM and light optics image work. TIA,
} We are interested in which lazer printers people have had experience } using for routine biological EM images. Our images are captured with a } Kodak Megaplus 1024x1024. We would also use the printer for LM, } anatomical line drawings, AR of gells etc. We are considering the } Lexmark Optra R+ 1200 dpi. Thanks for your help. } } Rick L. Vaughn } EM Research Facility } Dept. Cell Biology & Anatomy } Univ. Neb. Med. Ctr.
Hi Rick and Everyone, We have used the Lexmark Optra R printer in our lab for routine image printing for about 1 1/2 years and we WERE very happy with it. Recently, the high voltage regulator broke down and cooked a couple of toner cartridges. After talking to my refiller guy (been very knowledgable for years), it seems there is a longevity problem with these printers!! Several of his Lexmark clients have have opted (no pun intended) for another brand of printer. At this point he swears at them rather than by them. But, this is one man's opinion, and before I condem them completely, I would like to know if anyone else (or how many) has experienced this.
Cheers :) George Department of Earth and Atmospheric Sciences University of Alberta Edmonton, Alberta T6G 2E3 Canada ph: 403-492-5746 fax: 403-492-2030
We are beginning a series of experiments that will include labeling of = adipocytes for light microscopy, followed up by electron microscopy. = Preliminary experiments show that adipocytes are very autofluorescent, so = I was wondering if it is better to do immunoperoxidase staining when = working with these cells. If anyone is familiar with working with = adipocytes, and can give me any advice on which way is best to label = these cells, or if one block was found to be superior over another, I = would appreciate the information very much.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
To: Microscopy ListSer {Microscopy-at-MSA.Microscopy.Com}
I've got a set of these models on my FTP site, and everyone is welcome to them. I worked with Dave Joy in the mid '80's to convert these from Apple basic to IBM Basic. Later I took them from CGA to VGA resolution. They're compiled Quick Basic. The source code is not included, since I'm concerned about someone changing the code and effecting the validity of the results.
We have a Lexmark Optra Lxi well. It is installed on the network. I don't know how many people use it, but about a year ago, it was plugged in and turned on. The only maintenance I know has been to fill it with consumables.
------------------------------------------------- Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 Phone: (508) 750-1717 E-mail: crossman-at-osi.sylvania.com
Our web sites: www.sylvania.com www.siemens.com --
We have a Lexmark Optra Lxi well. It is installed on the network. I don't know how many people use it, but about a year ago, it was plugged in and turned on. The only maintenance I know has been to fill it with consumables.
------------------------------------------------- Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 Phone: (508) 750-1717 E-mail: crossman-at-osi.sylvania.com
Our web sites: www.sylvania.com www.siemens.com --
Hi, I need to count virus particles/ml of several viral suspensions with the viruses ranging in size from 20nm - 100nm. I need something quick and easy to give me a rough idea of the numbers. I've found some general information using polystyrene latex particles to do this, but I'd like to get a detailed protocol if anyone has one. 1. Do I need to get different sized latex particles for each virus size? The smallest latex I've found so far is 85nm. 2. Does anyone have a source for the latex? The companies I've called use them for mag. calibration and couldn't tell me how many particles/ml they contained. 3. Can I spray my sample on my grid with a nebulizer or should I drop it on? If I can spray it on, will I alter my count because of the extra water and PTA I'll be using?
Thanks for your help. Debbie Cassout TVMDL e-mail : dcassout-at-tamu.edu fax: (409) 862-7047
Can somebody please explain what Charge Contrast Microscopy is? I found this technique mentioned in a paper related to the analysis of ceramics, using a FEG-SEM. I was not successful in finding a description of this technique in my standard textbooks on microscopy or any of the microscopy related web-sites. Thanks for the help,
Hi Everyone, I am interested in attending an ultra microtoming class and I need information about a date, place and cost of an upcomming class if there is one. I work in the materials area, not biological. Thank you very much in advance, Jane Glamp glamp-at-ppg.com
Our larger o-rings usually come packaged with each o-ring having two or more overlapped turns. One of these o-rings must be inserted in a groove on the bottom of a plate in the scope, and I have not found a way to unfold the o-ring into a circle which will lay flat. I have put a book on it overnight and put it around a beaker slightly larger than the ID, but neither of those is satisfactory. I don't want to deform the o-ring, so I haven't been too rough with it. I am trying the beaker again, this time filled with hot water. (If the cold caused the o-ring on Challanger to harden, maybe the hot water will soften one.) Other than using so much grease that it holds the o-ring in place--obviously not good practise--I haven't found a way to keep the deformed o-ring in place. Has anyone out there solved this problem? TIA. Yours, Bill Tivol
Several people have asked me to post the replies I've received to my recent call for help...how to stain agar for acrylic resin embedding? I'm not going to post the complete replies - just a summary (lazy today). SO: *Fast green when the tissue is in 100% EtOH. Add a few ul of a 7% fast green solution to the vial with the samples. Doesn't work well with acetone. *Alizarin red for acetone dehydrations. *A light coat of india ink on the outsides of the agar blocks. *Dilute eosin (nobody ever says how dilute!). Acidified eosin. 1% Eosin B. *Alcian Blue. *2% Safranin O in the 100% EtOH step. *Mix some Dextran Blue in the agar.
There were also suggestions to fix with osmium or picric acid...both will color the material. Won't work for my samples, but...something to keep in mind.
Thanks to everyone who responded - I'm going to give the fast green a whirl today, since I have some on hand.
If anyone wants more info, I have the original messages and can hook you up with the people who sent them.
Hi Everyone, I am interested in attending an ultra microtoming class and I need information about a date, place and cost of an upcomming class if there is one. I work in the materials area, not biological. Thank you very much in advance, Jane Glamp glamp-at-ppg.com
You've probably heard from every vendor and his or her dog by now, and we also sell a variety of SEM specimen storage boxes, but, in my opinion, the *best* storage method is a good desiccator cabinet with shelving having holes to accomodate the specific specimen mounts you are using. At least two of the EM supply companies sell such systems, Energy Beam Sciences being one of them. My second choice, for standard "pin-type" specimen mounts, would be this lovely wooden mount storage box, made in the U.K., and sold by at least the same two vendors.
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
I have heard that organic diffusion pump oils could ignite under certain conditions. Can anyone confirm this or has anyone heard of such an event happening?
Ciao for now, Ken
Kenneth JT Livi Department of Earth and Planetary Sciences 34th and Charles Streets The Johns Hopkins University Baltimore, Maryland 21218
I count regurlarly virus particles with latex beads. You can use latex beads at a size between 90-140 nm at a concentration of about 10^8 beads/mL. You can find this beads with a known concentration at Marivac ltd, Halifax, Nova Scotia. (tl.902-429-0209). My protocol consist to mix 100 uL of viral suspension with 100 uL of latex beads in a 240 uL "Beckman Airfuge" tube. Place a grid in the bottom of tube. Centrifugate at 20 psi (about 120 000g) during 5 min. Take the grid, dry it and stain with phosphotungstic acid (PTA 3%, pH 6). I count on 2 differents grids at least 200 virus or beads in different areas.
Viral particle concentration (part/mL)= virus count / latex beads count X latex bead conc. X 1/dilution of test article
You can find my Airfuge technique in:
Alain R. and al., J.Vir.Methods, 16 (1987): 209-216
I don't have experience with nebulizer but I think it's a good conpromise.
Don't hesitate to write me if you want more details.
Robert Alain Microscopie Electronique Institut Armand-Frappier Laval, Quebec
Robert_alain-at-iaf.uquebec.ca
Hi, I need to count virus particles/ml of several viral suspensions with the viruses ranging in size from 20nm - 100nm. I need something quick and easy to give me a rough idea of the numbers. I've found some general information using polystyrene latex particles to do this, but I'd like to get a detailed protocol if anyone has one. 1. Do I need to get different sized latex particles for each virus size? The smallest latex I've found so far is 85nm. 2. Does anyone have a source for the latex? The companies I've called use them for mag. calibration and couldn't tell me how many particles/ml they contained. 3. Can I spray my sample on my grid with a nebulizer or should I drop it on? If I can spray it on, will I alter my count because of the extra water and PTA I'll be using?
Thanks for your help. Debbie Cassout TVMDL e-mail : dcassout-at-tamu.edu fax: (409) 862-7047
} Can somebody please explain what Charge Contrast Microscopy is? I found } this technique mentioned in a paper related to the analysis of ceramics, } using a FEG-SEM. I was not successful in finding a description of this } technique in my standard textbooks on microscopy or any of the } microscopy related web-sites. Thanks for the help, } } Hasso Weiland } } Alcoa Technical Center } Alcoa Center, PA 15068
Not a term I have come across. My first thought was EBIC - electron beam induced current - one step on from specimen current imaging, but both of these only apply to conductors or semiconductors.
As the specimen is an insulator, I guess the term is being used synonymously with voltage contrast. Again, mainly used for semiconductors but has been applied to ceramics. Non-conducting, or poorly conducting specimens charge if not coated. If everything is balanced out correctly, you can achieve a steady state where the charge input to the specimen equals the charge leakage to ground. Sometimes a very thin carbon coat, as used for EDX, is applied to help establish steady state conditions. In some specimens this is useful and contrast in images relates via the level of charge in different parts of the specimen to, among other things, local conductivity. Not very quantitative but quite sensitive. Check Goldstein et al (1992), Scanning Electron Microscopy and X-ray Microanalysis, 2nd Edition, Plenum Press, pp 557-562.
Regards,
--------------------------------------------------------------- Dr L. P. Stoter Technical Editor, MICROSCOPY & ANALYSIS Technesis 17, Rocks Park Road email: LPS-at-teknesis.demon.co.uk Uckfield, E. Sussex Phone: +44 (0)1825 766911 TN22 2AT Fax: +44 (0)1825 766911 United Kingdom ---------------------------------------------------------------
This is great! Ask and you shall recieve. In regards to George Braybrooks and Scott Whittaker's comments this makes two strikes against Lexmark. I didn't mention in my original message regarding lazer printers that we had tried several years ago using Lazer Master's card to double a HP4's resolution, but it took 50% of the RAM upon loading and was finicky with some hardware. Those of you that mentioned other cards: how are they for RAM usage and cost. Not to put down SEM but they seem to get by with non-photo printing easier than us with TEMs. An SEM print off our HP4 looks much better than the TEM?
I've taken on the responsibility of assembling separate short course and future meeting listings for the revamped journal of MSA, MAS and MSC: "Microscopy and Microanalysis," which will go to over 5,000 scientists.
If you have, or know of anyone who has, information concerning future short courses and/or topical meetings of interest to microscopists internationally, please send e-mail directly to me (not the listserver).
We will publish in date order short paragraphs giving Date and Title of the short course or meeting; Location; Name, phone, mail and e-mail ad- dresses of a contact person(s); and a short (50 word) description of the event.
The Jan/Feb issue is at press. The Mar/Apr issue closes this Friday, 24 January. If you wish to be listed in this issue e-mail (ascii format) me *TODAY* and I'll try my best to fit your contribution in.
Send me a e-mail for closing dates for the balance of the issues for 1997.
---------- } From: Jacky Larnould {larnould-at-mnet.fr} } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Asbestos counting } Date: Tuesday, January 21, 1997 1:35 PM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hi everybody, } And specially people working on asbestos. } I would like to know which method is used in the other countries (USA UK } Canada Germany....) to determine the concentration } of asbestos in atmosphere. } } Jacky Larnould } tel 33 (0)4 67 72 28 26 } fax 33 (0)4 67 79 54 90 } email larnould-at-mnet.fr } The following meyhods are currently in use for measuring airborne asbestos concentrations. 1. Phase contrast microscopy (PCM): U.S. and Britain for most occupational exposures 2. SEM: Germany, all exposure measurements 3. TEM: U.S. testing for airborne concentrations after asbestos removal operations in schools, for certain occupational studies and some public buildings.
PCM really gives total airborne fiber concentration because asbestos cannot be differentiated from any other fiber of similar size. SEM methods rely on EDS for identification but cannot differentiate asbestos from other mineral fibers with the same chemical composition but different crystalline structure. TEM uses both EDS and SAED and is the only method currently acceptable in law courts in the U.S.
The individual protocols for each are too involved to post on this server. If you are interested, contact me by email strangedoc-at-fuse.net and I'll try and help you further. K. A. Brackett, Ph.D.
From Microscopy-request-at-Sparc5.Microscopy.Com Tue Jan 21 18:45:42 1997 Mime-Version: 1.0
I received a lot of replying for my post on problem in zip disk. Many people said they have simlar problem. Finaly, I got my data recovered by Norton utility. It took very long time waiting the Norton to recognize the zip drive (it was keeping spin and click for 10+ minuts) and after recovering the files, the zip drive was completely damaged - any good-new disk will bdcame bad after insert to the drive. I called iomega then and got a return number to repair the drive. I'd like thank all of you who responded my question.
**************************** * Maoxu Qian, Ph.D. * * Dept of MSE, box 352120 * * University of Washington * * mxq-at-u.washington.edu * * (206)543-1514(phone) * * (206)543-3100(fax) * ****************************
} Hi, Nestor. Your ideas on plasma cleaning for specimens and stages are most } welcome -- good for you! The next question is .... can the cleaning be done } in an EM without damaging an EDX window (a thin one)? --------- stuff cut out ---------- } Cheers, } Brian } ==============
Brian etal....
I guess I'm not sure what you want to really do here. Do you want to just clean the EDS window, or if I'm guessing correctly the interior of the column? If it's the column then this is a good concept! Obviously once you clean the stage the next area to consider is the immediate surroundings which are also sources of contamination. Assuming of course your not in the situation where the vacuum system itself is poor and overwhelming the whole process aka vintage 70's microscopes. On the other hand, if we are talking about only cleaning EDS windows I would not recommend this be done with plasmas as the cleaning time will be far too long. Assuming we are talking about cleaning a column, then my comments below apply.
Basically, what you have to remember is that the plasma is acting like a catalyst for a localized (surface) chemical reaction. The energy of the plasma is breaking weak bonds of the hydrocarbon compounds on the surface which then make the species somewhat volatile so that they can further react with the gas in the plasma. The subsequent action of the gas is to provide an additional chemical process which is converting the compound into a gaseous phase which can be easily removed by the mild vacuum conditions. For example here are some reactions which could be used if we were dealing only with pure carbon...
C + 2H2 -} CH4 ; C + CO2 -} 2CO; C + O2 -} 2CO .....
Lots of other possibilities exist depending on the materials and "gas". If the EDS window compound is a strongly linked polymer then you will need a high energy plasma to disassociate the OH bonds to make an appropriate reaction proceed. This will occur only if you are running plasma's on the order of 50eV, or with very chemically reactive gas compounds (i.e. something more than just Ar, O2 ....). In this regime you can easily "etch" polymers and thus also a hydrocarbon based EDS window. If on the other hand you run with only very low energy plasmas ( { 10 eV) this should not be a problem. I would recommend you test the EDS window material first, as in most materials, some compounds are more resistant than others. If you can't do it locally just send some to me and I'll try it here for you in some of my gobs of spare time! ;-) My best guess is that there will be a readily set of conditions which will not cause problems for most hydrocarbon based thin-window detectors, however I have yet to test any here at ANL. The other obvious advantage is that the plasma is nearly a RT process. I've measured temperature rises in a thermocoupled TEM stage of ~ 5 degrees C under cleaning conditions, which is less than one gets using a 150W flood lamp!
If you decide to try this yourself, then I would recommend that you use a 2 step process first pure Ar then O2. The Ar disassociates the most volatile compounds first which then get swept away pretty quickly. The O2 finishes the job on the stuff that is more strongly bound. The presence of O-rings in the column will not be a problem, since the sealing surfaces are protected from the plasma. As an example consider the o-rings on specimen stages which survive multiple treatments in a stand alone table top system without problems.
Hope that this answers your question.
Nestor
BTW, to keep everything on the up and up, I will remind you all that the use of reactive gas plasma for cleaning the interior of an electron-optical column ( SEM, TEM, AEM...) is covered by an patent owned by ANL, who is my employer.
Dear Hasso, I have never heard that term, but last year I was looking at sintered mixtures of SiC and Al2O3. The SiC was dark, because it is conductive and the Al2O3 was white, as it was charging. This only worked if the mixture was more than about 20% SiC, otherwise the Al2O3 charged so much it disrupted the picture. In order to try to keep the charging down I tried doing very short gold-palladium sputter coats of 20 seconds or less, but even 10 seconds of coating completely obscured this contrast. The technique would only work with a well-dispersed mixture of conductive and non-conductive material. The proportion is easily determined by area percent image analysis. You wrote: } Can somebody please explain what Charge Contrast Microscopy is? I found } this technique mentioned in a paper related to the analysis of ceramics, } using a FEG-SEM. I was not successful in finding a description of this } technique in my standard textbooks on microscopy or any of the } microscopy related web-sites. Thanks for the help, } } Hasso Weiland } } Alcoa Technical Center } Alcoa Center, PA 15068
Regards, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
Dear Bill, When our EM sevice engineer wants to "plump up" an o-ring that has been in service for a long time, he puts it into an oven at 50 degrees C for an hour. This seem to relax them and plump them back to round. Perhaps if you put them in an oven with a weight on them for a while, then keep a weight on them as you cool them down, this would flatten them enough. You wrote: } Our larger o-rings usually come packaged with each o-ring having } two or more overlapped turns. One of these o-rings must be inserted in a } groove on the bottom of a plate in the scope, and I have not found a way } to unfold the o-ring into a circle which will lay flat. I have put a book } on it overnight and put it around a beaker slightly larger than the ID, but } neither of those is satisfactory. I don't want to deform the o-ring, so I } haven't been too rough with it. I am trying the beaker again, this time } filled with hot water. (If the cold caused the o-ring on Challanger to } harden, maybe the hot water will soften one.) Other than using so much } grease that it holds the o-ring in place--obviously not good practise--I } haven't found a way to keep the deformed o-ring in place. Has anyone out } there solved this problem? TIA. } Yours, } Bill Tivol } Regards, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
I'm doing an ALCHEMI (Atom Location by Channelling Enhanced MIcroscopy) study on a dopant in cubic zirconia. The literary references I've found to date mostly deal with intermetallics where atoms A and B (and impurity atom X) *can* lie on either the alpha or beta site of a structure (e.g., L1o superstructure).
My problem: I'd like to find a reference or two on an ALCHEMI study of an ionic material where atoms A and B can *not* reside on either of the sites (meaning, for example, that Zr would not be found on the O sites and vice versa). The mathematical treatments in the above references allow for this to occur and often times rely on an iterative convergence to the site occupancies of the X atom.
If you are aware of specific ALCHEMI studies on ionic (and perhaps covalent) materials, I'd appreciate the reference.
} Our larger o-rings usually come packaged with each o-ring having } two or more overlapped turns. One of these o-rings must be inserted in a } groove on the bottom of a plate in the scope, and I have not found a way } to unfold the o-ring into a circle which will lay flat. I have put a book } on it overnight and put it around a beaker slightly larger than the ID, but } neither of those is satisfactory. I don't want to deform the o-ring, so I } haven't been too rough with it. I am trying the beaker again, this time } filled with hot water. (If the cold caused the o-ring on Challanger to } harden, maybe the hot water will soften one.) Other than using so much } grease that it holds the o-ring in place--obviously not good practise--I } haven't found a way to keep the deformed o-ring in place. Has anyone out } there solved this problem? TIA. } Yours, } Bill Tivol
Bill,
Not a problem I've faced, but how about putting it around the beaker, as you've been doing and then putting it in the deep freeze over night. When it comes out nice and hard, you'll probably have a minute or so to get it into place and the bottom plate located, wait a few minutes for it to soften and then tighten the bolts. Worth a try? ... :)
I can confirm that heat for restoring o-rings does work in many instances. For many years I have successfully used hot water to do this rather than an oven.
Regards
Robin Cross EM Unit, Rhodes University, Grahamstown, South Africa
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa eurc-at-giraffe.ru.ac.za - tel: +27 461 318168 - fax: +27 461 24377
Matthew Stough wrote: } I'd like to find a reference or two on an ALCHEMI } study of an ionic material where atoms A and B } can *not* reside on either of the sites (meaning, } for example, that Zr would not be found on the O } sites and vice versa). } .... } If you are aware of specific ALCHEMI studies on } ionic (and perhaps covalent) materials, I'd appreciate } the reference.
Dear Matthew, Much of the early work on ALCHEMI by Spence and Taftoe was done on ionic materials, e.g. with spinel structure. You will find several references in
J.C.H. Spence & J. Taftoe, Journal of Microscopy, vol. 130, pt. 2, May 1983, p. 147 - 154.
The papers by Rossouw et al. on axial channeling ALCHEMI are also concerned with ionic materials of spinel and perovskite structures:
C.J. Rossouw, P.S. Turner & T.J. White, Philos. Mag. B vol. 57, 1988, p. 209 - 225.
C.J. Rossouw, P.S. Turner, T.J. White & A.J. O'Connor, Philos. Mag. Letters 60, 1989, p. 225 - 232.
Personally, I have had good experience with the axial channeling method.
Best wishes, Jorgen.
-at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- J. B. Bilde-Soerensen Materials Research Department Risoe National Laboratory DK-4000 Roskilde Denmark
} I guess that low humidity and constant temperature are the important } factors. I've thought (comments please) that it might be worth } replacing the air with an inert gas as well. } } Geoff Avern
Now that I've got email working again, I'll stick in my $0.02 worth. I agree with Geoff (and several others): humidty is very important. However, I'd rate protection against mechanical shock 2nd, then temperature, etc. Make sure the stubs are firmly mounted, sticky tape is only for temporary storage or for when you can be assured that specimens won't be moved. I've used specimens that I've shipped in a VW bug across country and ones shipped from Antarctica, and sticky tape would *not* have done the job. Stubs firmly mounted in commercial or specially made stub holders do OK. Also, make sure the specimens are firmly mounted on the stub--it can take only a small jar to knock specimens off the stub or de-orient them. Given the rough handling of specimens by many people, long-distance shipping isn't needed to lose samples, just a trip across the lab or campus. For medium to long term storage, vacuum (10-3) or *dry, oil-free* inert gas (N2 or Ar) is good; for archival storage, I'd exchange the air with N2 or Ar and then pump down. O2 can be a long term problem, but I wonder more about the general crud in the lab air. Phil
&&& Illigitimi non carborundum &&&&&&&& Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217)244-3145 days (217)355-3145 evenings oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
To: Microscopy ListSer {Microscopy-at-MSA.Microscopy.Com}
If you have a bulk N2 system as we do, gas boiled off the liquid is super clean. We're plumbed up to it for air tables, sample blow off, and we bleed it slowly into those big plastic boxes with the foam door seals, bolted to the wall. This is convenient, ultra-dry, and close enough to vibration free. We just use any sample box designed for 1/8" stud sample mounts.
I was following the a discussion in the Confocal archives around 3/95 = regarding deconvolution of images. It was said at one point that to = accurately deconvolve an image, the complete system must be known. = Given a simple transmitted light microscope system, ie objective, = coverslip, etc, does anyone know what exactly must be "known", how one = goes about getting that information, and is there software that can take = that information to deblur or deconvolve an image? Also, can it be done = to a digitized image?
Gatan offers the following short courses at $750 each. Tim Hughan Gatan, Inc.
Gatan's 1997 Schools for Electron Microscopy:
Digital Microscopy April 7-10, 1997
EELS Imaging & Analysis April 15-18, 1997
Specimen Preparation April 28-30, 1997
Digital Microscopy
This course introduces TEM, STEM, and SEM users to the capabilities of Digital Microscopy. On-line digital imaging facilitates the immediate viewing of samples and has made quantitative acquisition and analysis of electron micrographs and diffraction patterns possible. The course will explore the parameters necessary to optimize TEM, STEM, and SEM image acquisition, processing, and analysis for biological and physical sciences applications.
EELS Imaging & Analysis
The purpose of this school is to inform potential Gatan PEELS and Imaging Filter (GIF) users of the capabilities of parallel-detection electron energy-loss spectroscopy (EELS) and energy-filtered imaging, and to give current users the necessary theoretical background and hands-on training to get the most out of their PEELS or GIF systems.
TEM Specimen Preparation
This extensive, practical course has been designed to teach materials scientists the art and science of TEM specimen preparation. The course will concentrate on the various ion-milling techniques now available and will show how excellent TEM specimens can be produced from almost any material encountered. Special attention will be given to preparation of cross- sections through surfaces and interfaces. The materials used in the laboratory sessions will include semiconductors, metals, ceramics, and their combinations.
April 20-25, 1998 7th Frontiers of Electron Microscopy in Materials Science Conference Irsee Germany Contact: W. E. King, L-356, LLNL, Livermore, CA 94551 E-mail: weking-at-llnl.gov
Please visit the Frontiers of Electron Microscopy in Materials Science Conference website at
http://multiscale.llnl.gov/femms98
------------------ RFC822 Header Follows ------------------ Received: by quickmail.llnl.gov with ADMIN;22 Jan 1997 13:07:48 -0800 Resent-Date: Wed, 22 Jan 1997 13:07:48 -0800 (PST) Resent-From: king-at-cms1.llnl.gov Resent-To: wayne_king-at-quickmail.llnl.gov Received: from pierce.llnl.gov by cms1.llnl.gov with SMTP; Wed, 22 Jan 1997 13:07:48 -0800 (PST) Received: by pierce.llnl.gov (8.6.10/LLNL-1.18/llnl.gov-03.95) id NAA11053; Wed, 22 Jan 1997 13:07:46 -0800 Received: from popcorn.llnl.gov by pierce.llnl.gov (8.6.10/LLNL-1.18/llnl.gov-03.95) id NAA11043; Wed, 22 Jan 1997 13:07:44 -0800 Received: from 128.115.25.7 by popcorn.llnl.gov (8.6.10/LLNL-2.0) id NAA23371; Wed, 22 Jan 1997 13:07:35 -0800 Message-ID: {32E68199.46B4-at-llnl.gov}
on Wed, 22 Jan 97 18:29:00 Jane Glamp wrote:
} Hi Everyone, } I am interested in attending an ultra microtoming class and I need } information about a date, place and cost of an upcoming class if } there = is one. I work in the materials area, not biological. } Thank you very much in advance, } Jane Glamp } glamp-at-ppg.com
} Jane,
AMC Group has offered intensive hands-on workshops on Materials Ultramicrotomy, at basic and advanced levels, twice a year, on a regular basis since 1993. The schedule of the workshops for 1997 is:
Basic Materials Ultramicrotomy May 19-21, =9197 and Nov. 17-19, =91= 97 (Phoenix, AZ) Advanced Materials Ultramicrotomy May 22-23, =9197 and Nov. 20-21 (Pho= enix, AZ)
To receive detailed information about these workshops, please send me yo= ur complete mailing address. Thank you.
Rene E. Nicholas Marketing Director AMC Group (602) 949-4203 Fax (602) 473-9421 **************
AMC Group offers hands-on workshops on TEM Wedge-polishing, FIB-TEM Cross-sectioning, and Materials Ultramicrotomy on a regular basis. =20
Is anyone willing to share their experience in collecting nasal tissue from cynomolgus monkeys?
Perfusion techniques are not an option.
Exact sites of collection are not known to date. I assume we will be collecting from proximal and distal turbinates, and possibly septum.
I Would be interested in any references to methods of collection, fixation. Also would be interested in nasal comparative anatomy of the Rat vs. Monkey.
I was wondering if anyone has tried air-drying plant samples using Hexamethyldisilazane? If so, how successful was it? Our critical point drier is out of commission and I am searching for alternative methods to CPD. If anyone has any other comments or suggestions for drying plant material, I would greatly appreciate it.
Susan Carbyn Atlantic Food and Horticulture Research Station Kentville, Nova Scotia B4N 3R2 Canada
Many thanks to all those who replied to my recent questions about closed circuit cooling systems for electron microscopes. It has been interesting following the subject as, I believe, something so fundamentally simple shouldnt be as complicated as it would seem. I think part of the problem is many people seem to be using locally available products with tradenames that are unheard of elsewhere. It would seem that many of these products are either anti-corrosion or anti-microorganism but often not both. I believe the supplier of the electron microscope should be able to supply the name of a product that is globally available at a reasonable price, or at least supply the name of a product in your area. The additive we were recommended with our last microscope purchase is available to us (from Germany) at a considerable cost. In addition, the supplier of the product recommends regular replacement of the water in the systems, a job I certainly dont want to do.
My questions, just for interest really, to those with closed circuit cooling systems on their microscopes. - What is the name of the anti-corrosion agent you use? - What ratio do you use it at? - How do you replenish it and how often?
- What is the name of the anti-microorganism agent you use? - What ratio do you use it at? - How do you replenish it and how often?
- What is the name of the manufacturer of these products?
TIA,
Allan Mitchell.
Please send responses to allan.mitchell-at-stonebow.otago.ac.nz
Richard Lander, NZCS South Campus Electron Microscope Unit c/- Pathology Department Otago Medical School P.O. Box 913 Dunedin New Zealand. Tel. National 03 479 7301 Fax. National 03 479 7254
Message on behalf of Zyg Poczwa, Scientific Officer in diagnostic EM;
I am requesting help with trying to diagnose a bone marrow biopsy which shows tumor infiltration. The pathologists query whether or not it is megakaryoblastic and consequently would like Platelet Peroxidase (PPO) performed on it. Unfortunately, the only material I have is a Formalin fixed, Paraffin embedded bone marrow specimen.
Has anyone had any experience with doing platelet peroxidase on paraffin embedded tissue?
TIA
Zyg Poczwa.
Richard Lander, NZCS South Campus Electron Microscope Unit c/- Pathology Department Otago Medical School P.O. Box 913 Dunedin New Zealand. Tel. National 03 479 7301 Fax. National 03 479 7254
We generally get very poor results with plant tissue using HMDS } } } } } } } } } } } } } } } } } } } } } } } } } } } } } }
} } I was wondering if anyone has tried air-drying plant samples using } Hexamethyldisilazane? If so, how successful was it? Our critical point } drier is out of commission and I am searching for alternative methods to } CPD. If anyone has any other comments or suggestions for drying plant } material, I would greatly appreciate it. } } Susan Carbyn } Atlantic Food and Horticulture Research Station } Kentville, Nova Scotia B4N 3R2 } Canada } } Phone: (902) 679-5566 } Fax: (902) 679-2311 } } E-mail: carbyns-at-em.agr.ca } } ******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
} Does anyone have any experience with SEM of reconstituted rat tail collagen
} matrix? The collagen is laid down onto glass coverslips and then cells are
} plated on top of the matrix. I am interested in the collagen more than the
} cells, but can't get any definition of fibers. In some areas there is some
} definition, but the majority of the surface seems compacted with little or
} no definition. I have tried a variety of preparation conditions including
} different fixatives as well as cpd vs. freon for drying. The latter seemed
} to produce the best results, but there is still little definition of the
} matrix.
} I have sputtered for different times, but I still seem to have a problem of
} charging that eventually burns the sample no matter what kV I use. Does
} this
} sound like a preparation problem, an imaging problem, or both? Any help
} would be greatly appreciated.
} Thanks.
} Bill Swaim
Bill,
I have studied the conformation of fibronectin fibrils, which were formed either from fibroblast culture or in cell-free system, using a field emission SEM.
To succeed for high-resolution SEM, you need to preserve structures well in all specimen preparation steps and to reduce beam damage in the FESEM at high magnification. I use cryo techniques: fast-freezing, freeze-drying, cryo-transfer, cryo-coating, and cryo-SEM to achieve the goal.
The images obtained reveal the surface features of fibronectin fibrils and fibronectin molecule at a few nm resolution.
If you have any further questions or want to look at your samples, please feel free to contact me.
Here is a reference list:
Chen, Y., Centonze, V.E., Verkhovsky, A. & Borisy, G.G. (1995) Imaging of cytoskeletal elements by low temperature high resolution scanning electron microscopy. {italic} J.Microsc. {/italic} {bold} 179(1) {/bold} , 67-76.
Chen, Y., Brummel, S. & Peters, D.M.P. (1996) High resolution cryo-scanning electron microscopy in studying of macromolecular structures and assembly of fibronectin fibrils. {italic} Mol.Biol.Cell {/italic} 7(supplement),412a
Chen, Y., Brummel, S. & Peters, D.M.P. (1996) The structure and assembly of fibronectin fibrils studied by high resolution cryo-SEM. {italic} Scanning {/italic} {bold} 18(3) {/bold} , 200-201.
Chen, Y. & Peters, D.M.P. (1997) High resolution cryo-scanning electron microscopy (cryo HRSEM) study of the macromolecular structure of fibronectin fibrils. {italic} Scanning {/italic} , (In Press)
Peters, D.M.P., Chen, Y., Zardi, L. & Brummel, S. (1997) Conformation of fibronectin fibrils varies: discrete globular domains of type III repeats detected. {italic} J.Cell Biol. {/italic} (submitted)
Hope this answers your questions.
Regards,
Ya Chen
Ya Chen
*** My email address has been changed to: ychen14-at-facstaff.wisc.edu ***
We have not been successful in recovering peroxidase activity in parafin embedded tissue. The platelet peroxidase (as apposed to that in neutrophils) is very sensitive. Prolonged fixation will also remove activity. We have been successful at diagnosing M7 (megakaryoblastic leukemia) in peripheral blood using a combination of specific platelet peroxidase staining (using Breton-Gorius's published method) and immunostaining for GPIIb/IIIa (alphaIIb beta3 integrin). This is relatively easy to do and only requires that the peripheral blood have significant numbers of blasts.
Sorry I could not give a more positive answer, but with platelet peroxidase I think you will not be successful with anything other than very fresh tissue.
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-bgsm.edu * **************************************************************
On Fri, 24 Jan 1997, Richard Lander wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Message on behalf of Zyg Poczwa, Scientific Officer in diagnostic EM; } } I am requesting help with trying to diagnose a bone marrow biopsy which } shows tumor infiltration. } The pathologists query whether or not it is megakaryoblastic and } consequently would like Platelet Peroxidase (PPO) performed on it. } Unfortunately, the only material I have is a Formalin fixed, Paraffin } embedded bone marrow specimen. } } Has anyone had any experience with doing platelet peroxidase on paraffin } embedded tissue? } } TIA } } Zyg Poczwa. } } } Richard Lander, NZCS } South Campus Electron Microscope Unit } c/- Pathology Department } Otago Medical School } P.O. Box 913 } Dunedin } New Zealand. } Tel. National 03 479 7301 Fax. National 03 479 7254 } } "Southernmost EM Unit in the World!" } } }
} K. A. Brackett, Ph.D. wrote: } } "PCM really gives total airborne fiber concentration because asbestos cannot } be differentiated from any other fiber of similar size. } SEM methods rely on EDS for identification but cannot differentiate } asbestos from other mineral fibers with the same chemical composition but } different crystalline structure. } TEM uses both EDS and SAED and is the only method currently acceptable in } law courts in the U.S." }
The difference between the techniques is related to resolution and visibility in addition to the identification mentioned above. There are three common commercial types of asbestos: chrysotile, crocidolite, and Amosite. Single crystal chrysotile asbestos fibers (fibrils) are typically about 40nm wide, crocidolite asbestos has a larger width distribution but fibrils are commonly seen down to about 10-20 nm in width. Single crystals of Amosite are typically thicker than the other two commercial abestos types and may be a few tenths of a micrometer. (note: In all cases length is somewhat independent of the width, so that one can easily have very long-thin fibers.)
Chrysotile and crocidolite, therefore, can have a significant portion of the fibers below the resolution of the phase contrast light microscope (PCM). When analyzed directly on filters, the visibility (not resolution) of these thin fibers using a thermionic gun SEM is also very limited and not reliable for analysis. TEM can easily see/analyze the thinnest asbestos fibers. For Amosite, due to its thicker nature, the resolution/visibility case is more ambiguous and may be sufficient, especially in the SEM case.
Thus, PCM not only counts fibers indiscriminately (adding nonasbestos fibers), it also misses fibers indiscriminately (missing thin asbestos and other fibers). Thermionic gun SEMs have biases that are similar but not quite as severe as the PCM.
The reliability/use of the PCM/SEM/TEM also depends on its application. Asbestos occurs naturally in veins or mats of billions of bundled fibers. Since size fractionation of aerosol particles is a well known phenomenon, the sizes, especially the width, of the asbestos fibers/bundles in an aerosol will vary with location and the processes to which the asbestos has been exposed. Thus different locations/samples will have different size distributions. And different techniques (PCM/SEM/TEM) may get useful or totally useless results depending on the fiber sizes and types and the history of that particular aerosol.
To make the choice of method problem more complex yet ... only occupational (asbestos textile plants and the like) air samples analyzed by PCM have been correlated with health effect.
Again, as Brackett mentioned, the asbestos analysis issue can be very complex.
Eric B. Steel, Leader e-mail: eric.steel-at-nist.gov Microanalysis Research Group Office: 301-975-3902 N.I.S.T. FAX: 301-216-1134 Bldg. 222/Rm A113 Gaithersburg MD 20899
Matthew Stough wrote: } I'd like to find a reference or two on an ALCHEMI } study of an ionic material where atoms A and B } can *not* reside on either of the sites (meaning, } for example, that Zr would not be found on the O } sites and vice versa). } .... } If you are aware of specific ALCHEMI studies on } ionic (and perhaps covalent) materials, I'd appreciate } the reference.
Dear Matthew, To add on the reply from Joergen Bilde-Soerensen: There are two nice references from mineral physics:
Max T. Otten,1989, A practical guide to ALCHEMI, Philips Electron Optics Bulletin, 126, 21 - 28 (and refs therein)
Alex C. McLaren, 199, Transmission electron microscopy of minerals and rocks, chapter 7, Cambridge topics in mineral physics and chemistry, Cambridge University press, Cambridge
} Subject: Monte Carlo Models } } I've got a set of these models on my FTP site, and everyone is } welcome to them. I worked with Dave Joy in the mid '80's to } convert these from Apple basic to IBM Basic. Later I took them } from CGA to VGA resolution. They're compiled Quick Basic. The } source code is not included, since I'm concerned about someone } changing the code and effecting the validity of the results. }
David Joy's Monte Carlo models are also available, including source code in Borland Turbo Pascal ver 5, for example at
Many years ago I tried a variety of anti-corrosion, anti-bug additives, none of which were really satisfactory. On my current microscope, a Jeol 2000, I use distilled water without any additives. The water has not been changed in ten years, only topped up. In the absence of any nutrient bug growth is minimal and there have been no corrosion problems. Regards,
Eric Lachowski
---------------------- Dr Eric E. Lachowski University of Aberdeen Department of Chemistry e.lachowski-at-abdn.ac.uk
I am curious. After 28 years of changing tungsten filaments out in the sticks, and with little contact with the alternatives, what do you really think about LaB6 and FEG sources? This is in case we are successful in getting funding again!
Is FEG reliable or do you still get problems these days - I know in its early days there were some scare stories but how about now?
Any off-line/on-line comments would be appreciated.
Keith Ryan
Plymouth Marine Laboratory, Citadel Hill, Plymouth, Devon PL1 2PB, England
e-mail: k.ryan-at-pml.ac.uk
PML web site: http://www.npm.ac.uk/pml ++++++++++++++++++++++++++++++++++++++++++++++++++
} I am curious. After 28 years of changing tungsten filaments out in the } sticks, and with little contact with the alternatives, what do you really } think about LaB6 and FEG sources? This is in case we are successful in } getting funding again!
LaB6 has the great advantage that is has a life much longer than tungsten, therefore the money issue is maybe not so critical. Moreover you would not have to spend so much time in changing cathodes (a LaB6 cathode here lasts from 6 months to one year, to be compared with the 2 weeks for a tungsten one, if the microscope is used full time).
Also it must be emphazised that you get more intensity with LaB6, as well as more coherence of the beam, factors which may be critical for high resolution works.
The main problem may arise if the microscope is used by numerous users, when there is always a risk of someone doing something wrong. And it musr be noted that LaB6 needs "conditioning", meaning that it takes over one day (better a week-end) to have it ready to operate, if nothing gets wrong...
Yves MANIETTE Universitat de Barcelona Serveis Cientifico Tecnics Unitat ESCA TEM Carrer Lluis Sole i Sabaris, 1-3 E-08028 BARCELONA ESPANYA
Hi Susan. I keep an archive of biologic discussions posted to the microscopy list. There are two discussions which you may be interested in. Go to the web address listed at the end of this message and click on the "Tips & Tricks" link. From there go to the SEM section and look for the HMDS and Peldri links. Both should be informative. In answer to your question, HMDS might work. We teach an SEM short course twice a year and we make everyone dry their sample down with both HMDS and CPD to teach them the process and make them aware that different procedures can give different results. So far we have found no rhyme or reason to which plant tissues will work and which won't and we haven't made an attempt to catalog which do. I can't express an opinion on Peldri because we do not use it here. Boss man says it is a pain and that has been good enough for me. Hope this helps.
At 01:50 PM 1/23/97 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 352-392-1295 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
To: Microscopy ListSer {Microscopy-at-MSA.Microscopy.Com}
Difficulties in getting to my models on AOL have prompted me to move them. You should be able to FTP them from:
http://members.aol.com/dking99/mc
The self un-packing file is; MCVGAPAK.EXE Short instructions are in: MCVGAPAK.DOC
Again, these are based on Dave Joy's mid 80's Apple Basic model. Sorry for any inconvenience this has created. Please let me know how this works, what you think of the code, etc.
Message-Id: {2.2.32.19970124163047.006a3c34-at-po9.mit.edu} X-Sender: tonygr-at-po9.mit.edu X-Mailer: Windows Eudora Pro Version 2.2 (32) Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Keith Ryan asks about sources for EM's.
It all depends!!!
The only real advantage of a W source is that it is quite inexpensive to buy. Although changing a W source is relatively quick, I suspect the total down-time compared with LaB6 is comparable, because it has to be done quite often. The W source, of course, cannot begin to touch the performance of the LaB6.
LaB6 and FEG are now mature options. We have 4 LaB6 instruments and 2 FEG's. Premature gun failures are associated only with operator error. (In fact our FEG SEM is 2 years old and has not yet required a new tip, and the HB603 has run for 4 years on the same tip). Some people even question the need to heat LaB6 slowly these days, though we still do it. Our one remaining W microscope will be retired within the next few weeks, and replaced with another LaB6.
I hope this gives a sense of our opinions on the subject!
Tony Garratt-Reed.
**************************************************** **************************************************** ** ** ** Anthony J. Garratt-Reed ** ** Room 13-1027 ** ** Center for Materials Science and Engineering ** ** Massachusetts Institute of Technology ** ** 77 Massachusetts Avenue ** ** Cambridge, Massachsetts 02139-4307 ** ** U. S. A. ** ** ** ** Phone: 617-253-4622 ** ** Fax: 617-258-5286 or 617-258-6478 ** ** ** **************************************************** ****************************************************
We have a venerable LKB III Ultramicrotome in need of repair. The problem resides in the external control unit (swapping out another control unit from another LKB III solves the problem) and has to do with the specimen arm "quivering" rather than rising and falling in an appropriate manner. We have replaced vac tubes to no avail and now believe that professional assistance is warranted. Anyone know of a company or individual who might be of assistance? Thanks.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Southern Illinois University Carbondale, IL 62901-4402 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 ####################################################################
Susan Carbyn I did a small comparative study several years ago using HMDS, the now out of production Peldri, and CPD. I used renal and bronchial tissue with each treatment. The HMDS was as good as CPD for both tissues, but the fine cilia in the bronchial tissue looked better using the Peldri. I wish they had not taken that off the market. Another investigator working with whole fish (minnows) couldn't get any other drying procedure to work other than Peldri. For soft tissues, the HMDS seemed to have a greater chance of shrinkage artifact but this was not consistent. It did help to increase the number of infiltration steps from ETOH into 100% HMDS. I have heard that you can use Freon 113 as a drying agent but people I have talked to say they were not happy with it's results. Hope some of this helps instead of making it more confusing
Auburn University is seeking an electron microscopist who is experienced in biological electron microscopy and has a working knowledge of current techniques. A working knowledge of and experience with advanced light microscopic techniques are desirable. It is also desirable for the candidate to be able to apply electron microscopy to other fields.
Rank and Salary: Appointment is a non-tenure track research fellow position. The position is a 2-year appointment, renewable upon satisfactory performance and available funding. Salary is commensurate with training and experience.
Qualifications: Ph.D. in the biological sciences with an emphasis in electron microscopy.
Responsibilities: This position involves both research and teaching. The appointee is expected to manage and operate an electron microscope facility that has a Zeiss DSM 940 scanning electron microscope that is equipped with an X-ray unit, a Zeiss EM-10 transmission electron microscope, and ancillary equipment critical point dryer, vacuum evaporator, sputter coater, etc. Assist faculty, staff and graduate students in sample preparation and operation of the electron microscopes. Teach and/or assist in teaching courses in electron microscopy. Collaborate with reseachers and actively participate in writing research proposals and performing research. Write competitive research proposals and conduct original research.. Take the lead role in writing equipment proposals for upgrading the electron microscope facility. Interact well with users of the electron microscope facility and provide outreach to members of the community. Establish a working relationship with and attract funds from industry.
Application: Please send resume and names, addresses and telephone numbers of three
references to:
Christine W. Curtis, Chair, Search Committee Office of the Associate Provost and Vice President for Research 202 Samford Hall Auburn University, AL 36849-5112 (334) 844-4784 (Phone) (334) 844-5971 (Fax) curticw-at-mail.auburn.edu
Review of candidates will begin on February 15, 1997.
Auburn University is an Affirmative Action/Equal Opportunity Employer Minorities and Women are Encouraged to Apply
Columbian Chemicals Company has an immediate opening in the Physics Laboratory section of Laboratory Services located in Monroe, Louisiana. The job responsibilities include materials characterization and research in carbon black morphology and its interaction in vehicle systems (mainly elastomers) using scanning electron microscopy and energy dispersive x-ray analysis (SEM/EDS), scanning tunneling/multi-mode atomic force microscopy (STM/AFM), transmission electron microscopy (TEM), light microscopy and image analysis. We are seeking a candidate with a M.S. or Ph.D. in Carbon Science, Materials Science, Chemistry or Physics with a thorough background in one or several of the above microscopy techniques. Interested candidates should contact or send their resume to:
Dr. Alex Dmytraczenko Director, Laboratory Services Columbian Chemicals Company P.O. Box 96, Hwy 139 South Carbon Road Swartz, LA 71281 (318)329-8021
Columbian Chemicals Company is a subsidiary of Phelps Dodge Corporation and is the second leading produce of carbon blacks in the world with plants in North America, Europe, and Asia. Any interested candidates should be aware that Laboratory Services may be relocating to Marietta, Georgia in the near future.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have no idea how many hours are in the two weeks of W filament life but I suspect if it's life is that short you do not have a column vacumn that is low/high? enough for LaB6 or FEG source. Kate Connolly
A fellow researcher has been experiencing possible fixation/dehydration problems using wheat root tissue.
The method being used for our wheat roots is: fix in 2% paraformaldhyde plus 0.8% glutaraldehyde in 0.01 M Na-phosphate buffer overnight, followed by 3 changes of buffer, then 1% osmium for 1 hr, then 3 changes in water, then dehydrated (hold in 70% ETOH overnight), eventually embedding in LR White.
Main problems are:
1. Inconsistent infiltration of tissue
2. Very often get considerable shrinkage and distortion of the cortical tissue
Thank you for any suggestions,
Ginger Baker
Electron Microscopy Lab Manager Department of Anatomy, Pathology, and Pharmacology 250 Veterinary Medicine Oklahoma State University Stillwater, OK 74078 (405) 744-6765 FAX: (405) 744-5275 EMail: lizard-at-okway.okstate.edu
For 4 years RMC and the University of Arizona, Tucson, AZ have sponsored a workshop that has received excellent reviews from the participants, several of whom had attended other similar courses.
The course is typically held in October each year, the dates for this year will be set in February. We will post a second announcement at the time the course dates are set.
This course focuses on actual transfer of skills, not just knowledge. The instructors from each discipline of metals, polymers, thin films and biologicals all work with each student to truly understand the demands of their samples. Knowledge of glass knife making, diamond knife selection and care, types of resins, sectioning variables how to attack special problems are all part of the course.
For a complete prospectus please contact me at RMC, 4400 S. Santa Rita, Tucson, AZ 85714, phone 520-889-7900, fax 520-741-2200, email RMCBTLI-at-AOL.com
Steve Miller Director of Sales, North America Ultramicrotomy Division
At 08:30 24/01/1997 +0000, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America than 4 years with restriction due to a very good vacuum and the necessity to bake the gun once or two time a year. FEG Sem are now used by all people working in electron microscopy with no major problem, the resolution is very good even for low voltage, current is enough to do EDS (not WDS for now or in special condition with sealed counters...) and maintenance rather cheap (just consider the price of filament boxes during 4 of 5 years) FEG Tem are not very common for now because of cost but in term of analytical the machines are fantastic. Hope That helps.
Greetings from soggy California: I would like to know who is still making cryo-ultramicrotomes. With all the mega-mergers these past few years, I've lost track of all the "older" companies except for Reichert-now Leica. Are they still out there under different names? Thanks for your time.
John Hardy E.M. Facility City of Hope Med. Cntr. Duarte, CA (818) 301-8265 jhardy-at-smptlink.coh.org
In message {2E943120.1991-at-okway.okstate.edu} Ginger Baker writes: } To all: } } A fellow researcher has been experiencing possible } fixation/dehydration problems using wheat root tissue. } } The method being used for our wheat roots is: fix in 2% } paraformaldhyde plus 0.8% glutaraldehyde in 0.01 M Na-phosphate } buffer overnight, followed by 3 changes of buffer, then 1% osmium } for 1 hr, then 3 changes in water, then dehydrated (hold in 70% ETOH } overnight), eventually embedding in LR White. } } Main problems are: } } 1. Inconsistent infiltration of tissue } } 2. Very often get considerable shrinkage and distortion of the } cortical tissue } } Thank you for any suggestions, } } Ginger Baker } } Electron Microscopy Lab Manager, Department of Anatomy, Pathology, and } Pharmacology, 250 Veterinary Medicine, Oklahoma State University, Stillwater, } OK 74078, (405) 744-6765, FAX: (405) 744-5275 } EMail: lizard-at-okway.okstate.edu
Ginger,
I'd suggest boosting the glutaraldehyde concentration up to 2-2.5% for better fixation, and increasing the buffer concentratin to 0.1M. Your present buffer of 0.01M is quite low and you don't have much buffering capacity for roots of any appreciable mass. Also, extending dehydration and infiltration times may help. These may help eliminate the shrinkage and poor infiltration.
Under seperate cover, I'll send you technical tips on LR White proceedures that I've gleaned from this forum over the last few years (anybody else want them, please reply privately to my e-mail address).
Good luck!
Gib Ahlstrand, MMS Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
"When the mode of the music changes, the walls of the city will shake." - PLATO "There's a whole lotta shakin' goin' on!" - CHUCK BERRY
At 08:30 AM 1/24/97 +0000, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America The only thing I don't see having been mentioned here is your application. If you typically use 10 to 30kV, then there might not be any advantage of FEG over LaB6 unless you can afford the extra expense and want the ease of use. However, at voltages below 5kV you'll notice markedly improved brightness and improved resolution of the FEG over LaB6, and LaB6 over W.
And of course, by extra expense, we mean it isn't that expensive to add an ion pump to the gun region for LaB6, but for FEG you really ought to consider a different instrument.
cheers, shAf
{\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo ZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
Has anyone heard of "Wang Biomedical" microscopes ? Apparently thay are made in Holland. I am trying to find out about their reliability and quality.
Please send your responses directly to "cakyol-at-cisco.com" since I am not sure whether my membership request to this e-mail alias has been successfull or not.
In my Microscopy Vendors Database is all information about Wang Biomedical (postal address, phone, fax, e-mail and www address): http://www.kaker.com/mvd/vendors.html
At 11:34 AM 1/24/97 -0600, John J. Bozzola wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
John, We have three vintage LKB's (II and III) and would appreciate any info you obtain regarding service/repair. Rosemary Walsh
At 2:42 PM 1/24/97 -0800, Ginger Baker wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Ginger, You might trying leaving the samples under vacuum overnight during the primary fixation and then prolonging the infiltration steps. It depends on the specimen but you also might take the dehydration past 70% to 85% or further.
In response to all who are selling the idea of total digital photography. We are aware of digital technology, we have it, we use it, we love it. Heck if you have the cash digital image technology comes quite close to that of film. However, I still want to offer traditional darkroom capabilities as well, hence the request for a decent enlarger.
I am in agreement with the logic regarding the move from conventional darkroom to a totally digital lab. Going digital not only saves time in darkroom processing is more flexible, and also has environmental impact as well.
Currently our lab is 80% digital having digital capture from SEM, Image analysis, and partially from TEM. Currently we have two Kodak/Codonic 1600 series Dye sublimation printers. We also have decent CCD cameras, even use a Kodak DCS420 digital 35mm camera for other applications. Routinely we use digital images for publications, etc.
No doubt about it, digital photography has a strong future in electron microscopy and image analysis. However, I still believe there are occasions where traditional photography is the best alternative, especially since we have thousands of negatives in archives that on occasion need quality prints for our customers requesting that service.
Whatever the reason I believe we still have a need for darkroom enlargers for several more years.
For those who are fully digital, quite possibly you might be interested in donating your darkroom enlarger:-).
I would ask why you still need darkroom PRINT capability. With a good CCD with macro lens, you should be able to enlarge any area that you could using a photographic enlarger. However, I do not speak from direct experience, as I've never worked in a lab without a darkroom. However, I'm making a strong push to eliminate photopaper processing at my present place of employment. The basic point to be made is that a good dye sublimation printer can faithfully reproduce any captured image with proper grayscale rendition just as well as a photographic reproduction. Two years ago, however, I could not make the same claim. Thus, I would not recommed spending the $5-10K necessary to purchase a good new enlarger. Rather, invest the money in a high end CCD camera, such as the Kodak MegaPlus, and a decent dye-sublimation printer.
Has anyone heard of "Wang Biomedical" microscopes in Holland ?
I am trying to find out about their reliability, quality etc.
I am buying my first scope and any advice anyone offers will be much appreciated.
I tried to subscribe to the listserver but have not got a confirmation, therefore I'm not sure if I am included in the e-mail alias, so I would appreciate it if responses could be e-mailed to me: jakyol-at-pacbell.net
Gib, I would also be interested in receiving a copy of your accumulated LRWhite tips. Thanks in advance, Janet.
Janet K. Dye Ph. D. Graduate Student, Soils Land, Air, and Water Resources University of California Davis, California USA (916) 752-0199 (916) 668-4217 jkdye-at-ucdavis.edu
I am going to start looking at marine bacteria grown on plates and was wondering if someone had a good protocol for the fixation process. I will be doing SEM so I need to get rid of the salts and maintain cell integrity. I have used 2.5% glut in artificial sea water to fix but even with DH20 washes, I still get a lot of salt on the specimen and the specimen seems to be covered with a "slime" like coating. Is there a way to prevent this? I want to look at the alignment of the bacteria. Thanks for any help.
Peace through Christ,
Phil Rutledge, Director e-mail: prutle1-at-gl.umbc.edu Center for Microscopy voice: (410) 455-3582 UMBC Dept. of Biology fax: (410) 455-3875 Catonsville, MD 21228 ///// / / / / /////// /////// / / /////// /////// / / / / / / / / / / / / /////
I do not have experience with FEG (which I would love to have), but with LaB6, I believe that you can count on the following advantages, in order of importance:
1) Less "downtime" - much cleaner emission source. This also equates to less $$ in the long run, if you consider what your time is worth. 2) Higher EDS count rates. This means you can run at lower kV's and still get reasonable statistics and good analyses; almost a requirement for charge sensitive samples. 3) Higher brightness; again, you can run at lower kV's and get better S/N in your image.
BTW, depending upon your microscope, the cost of LaB6 is rather minimal - about $600. Might be worth the investment. I use one LaB6 source at a time, and use tungsten as my "backup". Works pretty well.
************************************ Bob Citron Chiron Vision 555 W. Arrow Hwy Claremont, CA 91711 USA Email: Bob_Citron-at-cc.chiron.com *************************************
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am curious. After 28 years of changing tungsten filaments out in the sticks, and with little contact with the alternatives, what do you really think about LaB6 and FEG sources? This is in case we are successful in getting funding again!
Is FEG reliable or do you still get problems these days - I know in its early days there were some scare stories but how about now?
Any off-line/on-line comments would be appreciated.
Keith Ryan
Plymouth Marine Laboratory, Citadel Hill, Plymouth, Devon PL1 2PB, England
e-mail: k.ryan-at-pml.ac.uk
PML web site: http://www.npm.ac.uk/pml ++++++++++++++++++++++++++++++++++++++++++++++++++
Message-Id: {3.0.32.19970127094222.00711b48-at-darkwing.uoregon.edu} X-Sender: mshaf-at-darkwing.uoregon.edu X-Mailer: Windows Eudora Pro Version 3.0 (32)
At 10:11 AM 1/28/97 +0400, fams-at-holonet.net wrote: } You actually write nothing, but attach 3 documents (30kb) ... I really do wish people wouldn't send attachments to any list. Please understand that we have to download them whether we want them or not, and if we choose to delete the e-mail (for whatever reason) we ^also^ have to find and delete the attachments. For me, this happens to be easy ... I know where attachments are saved ... but this is not generally true of everyone on a list.
Let me suggest you make the list aware of such announcements, and if anyone responds then you can send them the info ... or (better still) you could make us all aware that these announcements are available at a website ...
We have a penetron swirling shaker and over the weekend, something happened to it. The O-ring inside snapped, and this morning I got it temporarily replaced. Now, when I turn it on, it squeaks real loud! Apparently, it was making this noise over the weekend before it snapped. Does anyone have this type of shaker or have any ideas of how I could quieten this down? Perhaps there is something wrong with it! It is not that old, but I believe that the warranty has run out. I need to use it now but am afraid that I may be doing more damage by letting it run.
Any help or suggestions are welcome! Thanks, Susan
P.S. I want to thank everyone who replied to my HMDS question. I find this news group to be incredibly helpful! Thanks again.
Susan Carbyn Atlantic Food and Horticulture Research Station Kentville, Nova Scotia B4N 1J5 CANADA
At 10:10 AM 1/27/97, rutledge phil wrote: } I am going to start looking at marine bacteria grown on plates and was } wondering if someone had a good protocol for the fixation process. I will } be doing SEM so I need to get rid of the salts and maintain cell } integrity. I have used 2.5% glut in artificial sea water to fix but even } with DH20 washes, I still get a lot of salt on the specimen and the } specimen seems to be covered with a "slime" like coating. Is there a way } to prevent this? I want to look at the alignment of the bacteria. Thanks } for any help.
Phil,
We routinely use 2% glut in 0.1-0.2 M sodium cacodylate. With three washes, post-fix in OsO4, three washes, dehydration in ethanol, and CPD we usually don't have a problem with salt precipitation. The slime could be a mucous coat from the bacteria, therefore might need a prefixation treatment. If your preparation includes the plate media then that could be the problem.
A couple of references:
Watson, LP, AE McKee, and BR Merrell. 1980. Preparation of Microbiological Specimens for Scanning Electron Microscopy. Scanning Electron Microscopy. II: 45-56.
Krueger, DM, RG Gustafson, CM Cavanaugh. 1996. Vertical Transmission of Chemoautotrophic Symbionts in the Bivalve Solemya velum (Bivalvia: Protobranchia). Biological Bulletin. 190: 195-202.
Hope that helps, Louie Kerr
Louie Kerr Research and Education Support Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543 508-289-7273 508-540-6902 (FAX)
We have looked at settlement of marine bacteria (and whatever else) on glass plates, and I use my usual marine invertebrate fix:
4% glutaraldehyde in 0.1M cacodylate with 0.35M sucrose (2.5% glut will probably do)
Wash with 0.1M cacodylate with 0.44M sucrose
Postfix with 1% OsO4 in 0.1M cacodylate (sucrose optional)
Dehydrate with 30% - 100% EtOH as usual, then CPD.
Haven't had trouble with salt sticking around. Slime may or may not be removed - in many cases we WANTED to see the slime (but Murphy's law dictates that it will disappear if that's what we wanted to see). De-sliming seems to take place with thorough dehydration with EtOH (?).
Try to minimize the amount of culture medium that gets fixed onto the bacteria! That may be the culprit.
And remember to wear your lucky red shoes.
Tina
On Mon, 27 Jan 1997, rutledge phil wrote:
} Hi! } } I am going to start looking at marine bacteria grown on plates and was } wondering if someone had a good protocol for the fixation process. I will } be doing SEM so I need to get rid of the salts and maintain cell } integrity. I have used 2.5% glut in artificial sea water to fix but even } with DH20 washes, I still get a lot of salt on the specimen and the } specimen seems to be covered with a "slime" like coating. Is there a way } to prevent this? I want to look at the alignment of the bacteria. Thanks } for any help.
http://www.pbrc.hawaii.edu/bemf/microangelo **************************************************************************** * 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* ****************************************************************************
} Hi! } } I am going to start looking at marine bacteria grown on plates and was } wondering if someone had a good protocol for the fixation process. I will } be doing SEM so I need to get rid of the salts and maintain cell } integrity. I have used 2.5% glut in artificial sea water to fix but even } with DH20 washes, I still get a lot of salt on the specimen and the } specimen seems to be covered with a "slime" like coating. Is there a way } to prevent this? I want to look at the alignment of the bacteria. Thanks } for any help.
The slime is normal. Crang & Klomparens _Artifacts in Biological Electron Microscopy_ discusses this, and ways to avoid it. Avoiding sea salts is different. You might try washing with a sucrose solution adjusted to the same osmolarity as the sea water you're using, then washing with DDH2O. If you're not having problems with osmium precipitation after washing with sea water, you could do extended DDH2O washes after the OsO4--osmolarity is no problem (or less of one) after the Os. Phil
&&& Illigitimi non carborundum &&&&&&&& Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217)244-3145 days (217)355-3145 evenings oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
I am surprised that there have been so many comments from folks about different electron sources *in general*, without reference to the specific electron microscope Keith is using. LaB6 requires a better vacuum than tungsten (10 -6 torr or better) and FE requires a *much* better vacuum. Keith's EM must have this capability available to it for him to consider other sources than tungsten. As several folks have pointed out, the benefits also depend a great deal on the sort of work he is doing. The basic "rules of thumb" are that LaB6 is potentially 10 times brighter than tungsten, and that the cost of a source is about US$1.00/hour (these are very broad generalizations; your own "mileage" may vary...).
I would like to call attention to the fact that there is another option to be considered, even on an instrument with relatively poor vacuum: the use of a pointed tungsten filament to increase brightness.
Disclaimer: Energy Beam Sciences manufactures a range of proprietary pointed tunsgten filament tips for most EMs.
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
At 02:48 PM 1/27/97 -0500, Beth Richardson wrote: } Awhile back someone posted a message about a web site that listed used EM } equipment. Does anyone still have the address?
This information is available at the MicroWorld Resources and News web site, which is located at the URL:
http://www.mwrn.com
Best regards, Steven E. Slap ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
Our Website lists over 80 products and is fully hyperlinked to manufacturer's e-mail and websites. It is http://www.shore.net/~catalogs
Call if you would like a copy of the current catalog.
Regards,
Elinor Solit Director of Publications The Microscope Book
On Mon, 27 Jan 1997, Beth Richardson wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hi, } Awhile back someone posted a message about a web site that listed used EM } equipment. Does anyone still have the address? } TIA } Beth } } ************************************** } Beth Richardson } EM Lab Coordinator } Botany Department } University of Georgia } Athens, GA 30602 } } Phone - (706) 542-1790 } FAX - (706) 542-1805 } Email - beth-at-dogwood.botany.uga.edu } ************************************** } }
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
MSA Professional Technical Staff Award
The Microscopy Society of America (MSA) and the MSA Technologists' Forum are the sponsors of the Professional Technical Staff Awards (PTSA) to provide assistance on a competitive basis to full-time professional staff who submit papers for presentation at Microscopy and Microanalysis '97. The PTSA consists of free full registration for the meeting, a copy of the Proceedings and the Sunday evening social event at the Rock and Roll Hall of Fame. In addition, MSA will reimburse awardees up to $600.00 for travel, lodging and other expenses. It is the intent of this award to stimulate attendance for those who ordinarily might not participate, and to encourage employers to support their staff in professional activities. Applicants must have been full paid-up members of MSA for 3 years prior to the time of the meeting. Awards are based upon the quality of the paper submitted for presentation at the meeting. Abstracts will be judged by the MSA Technologists' Forum. The applicant must be the first author of the submitted paper. There will be four awards, two each in the Biological and Physical sciences. Successful applicants must present their papers personally at the Meeting in order to receive the award. Former winners will not be eligible for another award. Applications shall consist of (1) a photocopy of the completed Abstract and Data Form, originals of which can be found in the M&M '97 Registration Bulletin and Call for Papers, to be sent to the Technologists' Forum for judging on or before March 1, 1997. Judgment will be made and awardees notified to submit an original Abstract and Data Form by March 15, 1997. Those not receiving awards will also be notified in time to submit an Abstract and Data Form by March 15, if desired. (2) A supporting letter from the applicant's employer, manager or supervisor, attesting to the applicant's status as a full-time, professional staff member. Send a copy of the completed Abstract and the completed Data Form (copies only, no originals please), along with the supporting letter from your employer, manager or supervisor, to arrive by March 1, 1997, to Beverly E. Maleeff, Chair, MSA Technologists' Forum, SmithKline Beecham Pharmaceuticals, Toxicology-US, Mail Code UE 0462, 709 Swedeland Road, King of Prussia, PA 19406; Phone 610/270-7987; Fax 610/270-7202; E-mail Beverly_E_Maleeff-at-sbphrd.com.
} Subject: W, LaB6 and FEG sources } Author: Keith Ryan {KPR-at-WPO.NERC.AC.UK} at SMTP } Date: 1/24/97 8:30 AM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Keith,
After working with a good FEG, you will really like it. The FEG tip of our Hitachi S-900 in Madison was almost 10 years old. It was very stable, you can easily obtain a resolution test image at magnification of 350kx-400kx. That means that you have no down time for filament change, high brightness, high resolution. The key factor for FEG is a GOOD vacuum. We flash the tip every 2-3 days, that saves the life of FEG a lot. It was just replaced last week due to the extraction voltage went up.
Best regards,
Ya Chen
Ya Chen
========================================================================= \ / Integrated Microscopy Resource (IMR)-- \ / __ an NIH Biomedical Research Resource TEL : 608-263-8481 \/ / / University of Wisconsin-Madison FAX : 608-265-4076 / / / 1675 Observatory Drive #159 Email1:ychen14-at-facstaff.wisc.edu / /__/_ Madison, WI 53706 Email2:chen-at-calshp.cals.wisc.edu ========================================================================= IMR WWW Home Page: http://www.bocklabs.wisc.edu/imr.html
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have about $2300 to buy a writable CD system for a high end Mac = computer. Does anyone have any experience with any of these systems for = things like reliability, longevity, etc. We deal with lots and lots of = images and the Jazz and Zip drives get too expensive for the students = when storing many images. Thanks Judy M
Judy Murphy, PhD Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207
Phone: 209/474-5284 FAX: 209/474-5600 e-mail: murphy-at-sjdccd.cc.ca.us program web page: http://www.sjdccd.cc.ca.us/ElectMicro/sjdc.html
Carolina Workshop on: LIGHT MICROSCOPY FOR THE BIOMEDICAL SCIENCES June 1-6, 1997 University of North Carolina at Chapel Hill
Instructors: John J. Lemasters Edward D. Salmon Brian Herman ------------------------------------------------------------------- LIGHT MICROSCOPY FOR THE BIOMEDICAL SCIENCES is an introduction to applications of light microscopy. Students will have opportunities for extensive hands-on experience with state-of-the-art equipment for optical imaging, digital imaging processing, fluorescence microscopy and confocal microscopy guided by experienced academic and commercial staff. The course is divided into three major sections with lectures and laboratory exercises on: 1) geometric and wave optics of image formation, microscope alignment, phase contrast and reflection interference contrast microscopy; 2) video imaging, including contrast enhancement by analog and digital image processing, fluorescence microscopy, image detectors, fluorescent probes, ion imaging, and green fluorescent protein; and 3) laser scanning confocal microscopy emphasizing live cell imaging and 3-dimensional image reconstruction. Students are encouraged to bring their own specimens for analysis.
The workshop on LIGHT MICROSCOPY FOR THE BIOMEDICAL SCIENCES will cover basic concepts of light microscopy and introduce several advanced techniques relevant to modern cell and molecular biology. A commercial staff representing leading microscopic manufacturers will make available for student use the latest and most advanced instrumentation for light microscopy, image detection and computerized image analysis. Tuition is $950. ------------------------------------------------------------------- APPLICATION FORM Carolina Workshop on LIGHT MICROSCOPY FOR THE BIOMEDICAL SCIENCES
Position:
Address:
Telephone:
Fax:
Please return this form along with a brief letter describing your research interests and a curriculum vitae. Applicants should contact the program as soon as possible. Full consideration will be given to applications received by April 18, 1997.
Send application to: Dr. Wayne Litaker, Director of Workshops University of North Carolina at Chapel Hill Program in Molecular Biology & Biotechnology CB# 7100, 442 Taylor Hall Chapel Hill, North Carolina 27599-7100 Tel: (919) 966-1730 Fax: (919) 966-6821 e-mail: litaker-at-unc.med.edu ------------------------------------------------------------------- About Carolina Workshops: CAROLINA WORKSHOPS are intensive hands-on laboratory courses designed to teach cutting edge methods in molecular biology and biotechnology. Several courses on different topics in molecular biology and biotechnology are offered each year by the Program in Molecular Biology & Biotechnology at the University of North Carolina at Chapel Hill. Most participants in the Carolina Workshops already hold M.D. or Ph.D. degrees or are advanced pre-doctoral students. The courses are designed for novice students as well as for individuals with prior experience. All students benefit from in-depth interaction with instructors. ------------------------------------------------------------------- About the Instructors: John J. Lemasters, M.D., Ph.D. (Course Director): Dr. Lemasters is Professor and Director of Confocal Imaging in the Department of Cell Biology & Anatomy. Dr. Lemasters' research interests center on toxic and hypoxic injury, liver preservation for transplantation and mitochondrial calcium homeostasis, using confocal microscopy to monitor ions, membrane potentials, cell volumes, oxygen radicals and other parameters in single living cells.
Brian Herman, Ph.D: Dr. Herman is Professor and Co-Director of the Digitized Video Microscopy Facility in the Department of Cell Biology & Anatomy. Dr. Herman's research addresses the role of calcium, tumor suppressor genes, and anti-apoptotic proteins on regulation of cell growth and cell death using techniques of digital ion imaging, resonance energy transfer, confocal microscopy and fluorescence life time imaging.
Edward (Ted) D. Salmon, Ph.D: Dr. Salmon is a Professor in the Department of Biology whose interests are cell biology, cell motility, microtubules and mechanisms of mitosis and cell division. Dr. Salmon's research applies high resolution video and digital imaging microscopy towards understanding the molecular mechanisms governing the assembly of spindle microtubules and the segregation of chromosomes during mitosis. ------------------------------------------------------------------ Carolina Workshop on: LIGHT MICROSCOPY FOR THE BIOMEDICAL SCIENCES June 1-6, 1997 University of North Carolina at Chapel Hill
Instructors: John J. Lemasters Edward D. Salmon Brian Herman ------------------------------------------------------------------- {End of Announcement}
Yesterday I posted a message about the fixation of marine bacteria. I guess I should have added that the bacteria is being grown on agar in 9cm petri dishes. What I want to look at is the association of the bacteria to each other without disturbing the colony. There seems to be a particular orientation and I want to observe this by SEM without taking the bacteria off of the medium. Any suggestion? I appreciate all of the help so far, many thanks.
Peace through Christ,
Phil Rutledge, Director e-mail: prutle1-at-gl.umbc.edu Center for Microscopy voice: (410) 455-3582 UMBC Dept. of Biology fax: (410) 455-3875 Catonsville, MD 21228 ///// / / / / /////// /////// / / /////// /////// / / / / / / / / / / / / /////
} Date: Fri, 10 Jan 1997 08:26:57 -0700 } To: geoffa-at-amsg.austmus.gov.au (GeoffA) } From: george.braybrook-at-ualberta.ca (George Braybrook) } Subject: mucus removal } Cc: Microscopy-at-Sparc5.Microscopy.Com } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
I know this is old mail but can you explain why EDTA should not be used to= =20 decalcify specimens with bone?
Bob Schmitz
}
rschmitz-at-uwspmail.uwsp.edu or rschmitz-at-macsrv1.uwsp.edu=20 (note its macsrv"one" not "el") Robert (Bob) J. Schmitz Department of Biology,=20 University of Wisc. Stevens Point. Stevens Point, Wisconsin 54481 ph 715-346-2420
} Yesterday I posted a message about the fixation of marine bacteria. I } guess I should have added that the bacteria is being grown on agar in 9cm } petri dishes. What I want to look at is the association of the bacteria } to each other without disturbing the colony. There seems to be a } particular orientation and I want to observe this by SEM without taking } the bacteria off of the medium.
You should be able to process them _in situ_ by puddling the solutions on the colonies, exchanging them by careful pipetting. After drying, dissect away sample areas and thin the underlaying agar. Leaving the contents of the dishes intact in the dishes during processing should reduce or eliminate distortion/curling during drying. This assumes drying from HMDS. For CPD, dissect under 100% EtOH. Phil
&&& Illigitimi non carborundum &&&&&&&& Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217)244-3145 days (217)355-3145 evenings oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
} I have about $2300 to buy a writable CD system for a high end Mac } computer. Does anyone have any experience with any of these systems for } things like reliability, longevity, etc. We deal with lots and lots of } images and the Jazz and Zip drives get too expensive for the students when } storing many images. } Thanks } Judy M
Judy, This may be a case where you want to wait a year. The DVD discs are starting to come out now, with the 2nd generation late this year or next year. Don't get 1st generation--the standards for F2 is already known and incompatible with F1. DVD will likely replace CD-ROMs in a few years. Phil
&&& Illigitimi non carborundum &&&&&&&& Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217)244-3145 days (217)355-3145 evenings oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
} } I have about $2300 to buy a writable CD system for a high end Mac } } computer. Does anyone have any experience with any of these systems for } } things like reliability, longevity, etc.
{SNIP} } Judy, } This may be a case where you want to wait a year. The DVD discs are } starting to come out now, with the 2nd generation late this year or next } year. Don't get 1st generation--the standards for F2 is already known and } incompatible with F1. DVD will likely replace CD-ROMs in a few years. } Phil
Can you please explain what DVD media is please......are they comparable to mini-discs?
Thanks, Adam. ''~`` ( o o ) _____________________.oooO--(_)--Oooo._______________________________________ Adam Vivian-Smith PhD Student CSIRO/ University of Adelaide Voice: +61 08 8303 8627 Division of Horticulture Fax : +61 08 8303 8601 Urrbrae, Adelaide Email: Adam.Vivian-Smith-at-adl.hort.csiro.au S.A., 5064 AUSTRALIA .oooO ( ) Oooo. ________________________\ (____( )_________________________________________ \_) ) / (_/
Susan Carbyn wrote: ================================== We have a penetron swirling shaker and over the weekend, something happened to it. The O-ring inside snapped, and this morning I got it temporarily replaced. Now, when I turn it on, it squeaks real loud! Apparently, it was making this noise over the weekend before it snapped. Does anyone have this type of shaker or have any ideas of how I could quieten this down? Perhaps there is something wrong with it! It is not that old, but I believe that the warranty has run out. I need to use it now but am afraid that I may be doing more damage by letting it run. ================================================= SPI Supplies has been selling this product for roughly fifteen years and this is the first time I have heard of this (actually any) problem with it. Also, it is hard to diagnose just what the problem is without seeing the unit. However since this is perhaps about the most expensive shaker of this type money can buy, it would certainly be worth fixing. Let us know if you would like us to take a look at it. Unless the unit has been dropped, I am sure it would be more than worth repairing.
Chuck
====================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ =====================================================
} We deal with lots and lots of } } images and the Jazz and Zip drives get too expensive for the students when } } storing many images.
We had the same problem with students and buy a Sony CD-R (2X). The CDs does not cost so much (About $10). The price of the CD-R was $1000 for about 1 year ago.
Gary...
On Tue, 28 Jan 1997, Philip Oshel wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } I have about $2300 to buy a writable CD system for a high end Mac } } computer. Does anyone have any experience with any of these systems for } } things like reliability, longevity, etc. We deal with lots and lots of } } images and the Jazz and Zip drives get too expensive for the students when } } storing many images. } } Thanks } } Judy M } } Judy, } This may be a case where you want to wait a year. The DVD discs are } starting to come out now, with the 2nd generation late this year or next } year. Don't get 1st generation--the standards for F2 is already known and } incompatible with F1. DVD will likely replace CD-ROMs in a few years. } Phil } } &&& Illigitimi non carborundum &&&&&&&& } Philip Oshel } Station A } PO Box 5037 } Champaign, IL 61825-5037 } (217)244-3145 days } (217)355-3145 evenings } oshel-at-ux1.cso.uiuc.edu } *** looking for a job again ****************** } }
To: george.braybrook-at-ualberta.ca Cc: microscopy-at-Sparc5.Microscopy.Com
Phil, regarding your statement...
} This [i.e., buying a writable CD drive] may be a case where you want to } wait a year. The DVD discs are } starting to come out now, with the 2nd generation late this year or next } year. Don't get 1st generation--the standards for F2 is already known and } incompatible with F1. DVD will likely replace CD-ROMs in a few years.
...but will the drives be able to *write* (not just read) the current CD standard? If you want to distribute info widely (rather than just archiving), which Judy apparently wants to do, you don't want to do it on DVD until most people have access to drives that can read them.
On Tue, 28 Jan 1997, Robert Schmitz, Biology wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } EDTA fell out of favour as a decalcifier some time ago. } } I know this is old mail but can you explain why EDTA should not be used to } decalcify specimens with bone? } } Bob Schmitz } I'd like to konw the answer to this too, as we use EDTA to decalcify bone of the otic capsule routinely in inner ear histology. However, we are usually not interested in the bone itself, but the tissues it encapsulates.
Karen Pawlowski Inner Ear Histology UT Dallas/UT Southwestern Med. Ctr.
The successful candidate will be a analytical lab tech in the FAB-12 Materials Lab. Responsibilities will include supporting materials characterization for tool quals, process quals, process troubleshooting, reliability and manufacturing yield improvement. Primary responsibility will be hands-on support for TEM and SEM characterization requirements.
Skills:
Excellent communication/interpersonal skills required. The successful candidate should be able to prioritize multiple tasks to effectively meet customer needs in a changing environment. Candidate should have hands on experience with TEM sample preparation equipment used in dimpling/ion-milling or wedge polishing techniques. Experience with SEM and FIB are also highly desired. Minimum AA/AS degree or equivalent with 3-5 years work experience in materials analysis related to the semiconductor industry.
Intel is an equal opportunity employer. Resumes accepted by fax, email, or snail mail. Interested candidates should contact:
David Howell FAB 12 TEM Engineer Intel Corporation M/S OC2-218 4500 S. Dobson Rd Chandler, AZ 85248-4906 dave_howell-at-ccm.ch.intel.com Fax (602)715-8363
Dear All, Thanx for all the input. Here is a summary of that input, info from Parker and my own observations.
The most often-mentioned method to flatten o-rings was boiling. This method worked very well with viton. The advantages of boiling are that the method is easy to do reproducibly, and "It also drives out a lot of 'glup,' to use the technical term. (J. Pawley)". The disadvantage is that the water must then be removed. For viton, this is pretty easy, since that elastomer can be baked out at ~200 C. Another popular method was heating to 50 C in an oven. This was not quite high enough for viton, which did not lay flat after an overnight heating at that temp. I'm sure that heating at a somewhat higher temp would do it, however. The advantages of the oven method are that one need not remove water, and it is the best-controlled method for those elasto- mers, such as buna-N and fluorocarbons, which cannot be heated above 70 C. Following the heating process, one can cool the o-ring either slowly or rapidly. Slow cooling works for me, but there may be situations where shock cooling would be advantageous. In particular, to shape an o-ring to a particular non-flat or non-round configuration, it might be good to heat, shape, then shock-cool. The opposite suggestion--to put the o-ring around a beaker and freeze it in the round state--would not be applicable for my purpose. The reassembly of the column takes so long that the o-ring would thaw and fall out; furthermore, there would be condensation. I can imagine situa- tions where it could be useful to shape an o-ring, freeze it, and install. Spring clips were also suggested for holding the o-ring in place. This also would not be suitable for my situation, but might be useful to consider. A caution about silicone o-rings was that they are very permeable to He. As a result, leak-checking can give false positives for several days. Both viton and silicone o-rings can be baked out at ~200 C, and that may be a good idea for a standard practise, since it will drive off volatiles in the o-rings. Buna N and fluorocarbon cannot be heated above 70 C, and that for only a few hours. Buna N just melts, but fluorocarbon decomposes. Ethylene-propylene is the most radiation-resistant of the common elastomers. We use it for seals which are close to the beam, and it remains relatively flexible under circumstances where either viton or neoprene harden. The Parker O-Ring Handbook is a useful source of info about many properties and applications of various available elastomers. Since elastomers are treated by crosslinking and with additives, and are, no doubt, optimized for particular applications, the appropriate temps and times for particular treatments should be determined experimen- tally, rather than relying on info from books. Some info--such as decom- position temps--can be obtained reliably from books and used to set upper limits for trial runs, but even in this case, it is probably better to talk to the manufacturer before approaching these limits, since the treat- ments may significantly change them.
As usual, the list was a great source of info. Thanx, Nestor, for establishing and maintaining it. Yours, Bill Tivol
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Phil, regarding your statement...
} This [i.e., buying a writable CD drive] may be a case where you want to } wait a year. The DVD discs are } starting to come out now, with the 2nd generation late this year or next } year. Don't get 1st generation--the standards for F2 is already known and } incompatible with F1. DVD will likely replace CD-ROMs in a few years.
} } ...but will the drives be able to *write* (not just read) the current CD } } standard? If you want to distribute info widely (rather than just } } archiving), which Judy apparently wants to do, you don't want to do it on } } DVD until most people have access to drives that can read them.
} } Alfred
The issue of access to read the disks is why we decided to use CD-R's instead of Zip drives or other storage media. Nearly everyone has a CD player for retrieving data and the one recorder (Optima 650) can be moved around for recording. We are fairly happy with this recorder but it seems to me that I get more recording errors than I would like (2 or 3 image files per 50 need to be individually recorded or otherwise manipulated, but these are mostly Photoshop files and these problems may be confined to that type). Not having wider experience with CD-R's I don't know if it is more or less than typical.
Can anyone help locate a proceedure for staining a whole chick embryo with Alizarin Red? We did find something for sectioned material (Dahl's Method for Calcium) but wonder if there is a specific protocol for whole tissues, perhaps with a tissue clearing step. Thanks, Linda Fox lfox1-at-wpo.it.luc.edu Loyola University Medical Center Maywood Illinois
Concerning the question about ignition of organic diffusion pump oils:
Most organic compounds will ignite if heated to a sufficiently high temperature, and there are specific tests designed to characterize this property. It has been more than 50 years since I worked on such matters, but as I recall the most common test involves heating the fluid in an open dish over a bunsen burner, in a well-specified configuration, and noting the temp at which it catches fire. Most manufacturers provide flash point data in the literature for their DP oils. Here are values I found in my files for a few common DP oils: Convoil-10 190 C; Convoil-20 217 C; Octoil-S, 209 C; DC-704, 221 C; DC-705, 243 C; Alcatel 22, 280 C; Santovac-5, 288 C; Neovac SY, 230 C; the perfluorinated Krytox and Fomblin fluids, completely non flammable. Interestingly, these values are comparable to the boiling temperatures for these fluids at a pressure of about 100 Pa, where most DPs operate (see Vacuum Methods in Electron Microscopy, p. 181); however, I have never heard of the oil in a DP igniting under ordinary (and even some rather extraordinary) conditions of use and misuse. Even if it did, the fire would be relatively well confined and could be easily extinguished by placing something over the throat of the pump to exclude air.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
I have not been aware that corrosion (i.e. the dissolving away of metal in an aqueous medium) was much of a problem in the cooling systems of most electron mocroscopes and related instruments. It was my impression that such suystems are usually constructed of metals such as stainless steel and copper, which are quite resistant to corrosion under ordinary operating conditions. This has certainly appeared to be the case for the dozen or so instruments I have dealt with over the last several decades. That is, all we have done is to use ordinary distilled water (which we usually bought from a local drug store or grocery store, where it is stocked for people to use in steam irons) in our recirculating cooling systems, and we did this mainly to reduce the formation of mineral deposits, such as might result from the use of ordinary tap water. If you are experiencing evidence of corrosion, such as having the water become 'rusty', I would recommend that you check to see if someone has installed an ordinary steel coupling or other fixture somewhere in the system where it is in contact with one of the more inactive metals such as Cu or StSteel. In that case such metal-to-metal contact would form a galvanic cell that would lead to fairly rapid corrosion of the ordinary steel part. All you should need to do is to replace this steel part with a comparable part made of the metal with which it is in contact. This would eliminate the galvanic cell and the corrosion.
Control of the growth of algae is discussed in some detail in 'Vacuum Methods in Electron Microscopy' (p. 216). As noted there, we have had good success using the compound Chloramine-T (the sodium salt of N-chloro-p-toluenesulphonamide) at the level of about 0.25 gram per liter. This compound is not exceedingly expensive and is commonly available from specialty chemical companies such as Polysciences, Aldrich and Sigma. Excluding light from all parts of the circulating system also helps supress algal growth (i.e. don't use transparent plastic tubing). We have only changed the water when it looked dirty or happened to develop noticable amounts of algal growth. Others have recommended controlling algal growth by: adding enough sodium borate to raise the pH to a value of 9 (making the water alkaline in this way would also supress corroison of iron parts); the use of Dichlorophene (2-2-methylenebis-P-parachlorophenol); and the use of a compound called 'Aqua Treat', available from Aqua Labs (508-388-3989). I have had no direct experience with these latter three methods. Good luck,
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
I seem to remember that in December there was a group of replies to someone who asked if it was possible to prepare bone for TEM without decalcifying it. As luck would have it, I now have a P.I. asking about that very subject. From what I remember, my impression was that it was possible to prepare bone samples without any special procedures being needed. Did I remember correctly?
I am also interested in staining for calcium. My subject is Douglas Fir ectomycorrhizal with a basidiomycete fungus, Rhizopogon vinicolor. I am trying to localize calcium in the fungal tissue and in the interface area between the plant and the fungal cells. Tracers for calcium have proven difficult, i.e. flourescent probes for calcium do not cross fungal membranes and are probably too large to pass through cell walls as calcium does. I am interested in references on Alizarin red, or "Dahl's method for Calcium", whether these are applicable to botanical specimens, and what about fixing the calcium in place so that it is not displaced by sectioning, etc., prior to staining? Thanks in advance for any advice.
Janet K. Dye Ph. D. Graduate Student, Soils Land, Air, and Water Resources University of California Davis, California USA (916) 752-0199 (916) 668-4217 jkdye-at-ucdavis.edu
I'd recommend going with a CD-writer too, for the following reasons;
1) Your clients (students, faculty members, etc) are MUCH more likely to have immediate access to a CD reader than a DVD.
2) CD standard won't dissappear in a hurry because the whole music industry is locked in on it. It's probable that DVD's will read 'old' CD's too.
3) Assuming a micrograph is around 1MB in size, a CD will hold approx. 600 micrographs - more than enough already. Why would a student, working on your average sized project, want a DVD that holds 4,000-20,000 micrographs? Of course it's a different matter for archiving within the EM lab.
4) I don't know the cost of a DVD witer, but I bet it's a LOT more than a CD writer. You've got the money for a CD writer now. Why wait til you can afford a DVD?
Geoff Avern Microscopy Labs Australian Museum Sydney, Australia
Philips CDD 522 - we're very happy with it. (Usual disclaimer)
} Can you please explain what DVD media is please......are they comparable to } mini-discs? } } } Thanks, Adam.
DVD is the new standard for multigiga byte storage. CD-ROMs with multiple layers of data. The 1st generation is about 4GB, the 2nd about 9GB. I believe they'll be about the same diameter as CDs. Not comparable to mini-discs. Phil
&&& Illigitimi non carborundum &&&&&&&& Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217)244-3145 days (217)355-3145 evenings oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
At 08:58 29/01/1997, Alfred Kracher wrote: } Phil, regarding your statement... } } } This [i.e., buying a writable CD drive] may be a case where you want to } } wait a year. The DVD discs are } } starting to come out now, with the 2nd generation late this year or next } } year. Don't get 1st generation--the standards for F2 is already known and } } incompatible with F1. DVD will likely replace CD-ROMs in a few years. } } ...but will the drives be able to *write* (not just read) the current CD } standard? If you want to distribute info widely (rather than just } archiving), which Judy apparently wants to do, you don't want to do it on } DVD until most people have access to drives that can read them. } } Alfred
True. As far as I know, the 1st generation of DVDs will be read-only or if writable, then only to 1st gen DVD. 2nd gen DVD will read & write only 2nd gen DVD. Current CD-ROM will only read & write to current CD-ROM. Except probably not to all current CD-ROM; as they go to 12X (maybe even 8X) they're changing what they hold constant: the angular velocity or the data density. Currently CD-ROMs change speed as they read from inside=} out. The "faster" ones hold the rotation speed constant, as I recall. Anyway there is/will be compatibility problems between {6-8X CD-ROMs & (8?) 12X and higher ones. Phil
&&& Illigitimi non carborundum &&&&&&&& Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217)244-3145 days (217)355-3145 evenings oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
} Can anyone help locate a proceedure for staining a whole chick embryo } with Alizarin Red? We did find something for sectioned material } (Dahl's Method for Calcium) but wonder if there is a specific } protocol for whole tissues, perhaps with a tissue clearing step. } Thanks, Linda Fox
Linda, _Staining Procedures_, 3rd ed. Pg.137ff. Biological Stain Commission, Williams &Wilkins Co. Baltimore. 4th ed. has it also, but don't know the page. This work is likely on a higher edition by now. Email me if you need the detailed recipe. Hi to John! Phil
&&& Illigitimi non carborundum &&&&&&&& Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217)244-3145 days (217)355-3145 evenings oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
This is possibly a very dumb question, but anyway:-
If the carbon coating on a sample is imperfect, or if the earthing from said coating is imperfect, so that the sample charges, is the resulting charge positive or negative? My initial assumption was that is would be -ve (buildup of all those bombarding electrons with nowhere to go), but then surely one 15kV electron produces more than one secondary, doesn't it? I've recently had an incident whereby I got EPMA analytical totals which were a bit high (101 to 103), and this went away when I was more liberal with the conductive paint. Could this be because the sample was charging +ve, thus increasing the effective accelerating voltage?
cheers
Ritchie
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
The Alizarin technique for calcium is called "Dawsons" and is quite old. We don't have the original reference, but for bone the proceedure is: fix in 10% NFS, wash then transfer to 0.5% aq KOH to which enough alizarin red S has been added to turn solution to deep purple, leave for 24 hours or till bones are distinctly red, transfer thro KOH-glycerine series,3:1, 1:1, 1:3, to pure glycerine. Store in pure glycerine plus few crystals of thymol. Refs for meth blue plus alizarin red staining of bone may help: Bechtol 1948 Stain Technol 23, p3-9; Burdi and Flecker 1968 Stain Technol 43, p47-48; Lundvall 1927 Anat Anz 62, p353-373. If you still require more info, contact me via e-mail and we will try to help.
Miss A.J.Wilson Electron Microscope Unit St George's Hospital Medical School Cranmer Terrace Tooting London SW17 ORE Tel: 0181 725 5220
We found Dawson's original technique for alizarin preparations:
1. Fix in 95% ALC for 48-72Hrs. Prolonged fix in alcohol renders tissue less liable to maceration.
2. Remove fats in acetone for 2-4 days then return specimens to alcohol for 12-24 hrs.
3. Place in 1% KOH until bones or digits appear thro' muscle Transfer to 0.1% Alizarin Red S in 1% KOH. (Other stains eg alcian blue, or victoria blue can be used to counter-stain cartilage or bone at this stage.)
4. Leave till stained or desired bone colour, change to fresh stain if necessary.
5. CLEARING Place in solution of 1g KOH, 20mls glycerine, 79mls dist water, leave till sample clears.
6. When clear, pass thro' increasing conc of glycerine, 50, 70, 90, 100%
Miss A.J.Wilson Electron Microscope Unit St George's Hospital Medical School Cranmer Terrace Tooting London SW17 ORE Tel: 0181 725 5220
The "Advertisement For Applications Engineers" has an unreadable attachment. It appears to be a Wordpro document which I cannot handle. Please repost the message without using attached files.
} This is possibly a very dumb question, but anyway:- If you don't know the answer and you want to know it, then no question is dumb.
} My initial assumption was that is would be -ve (buildup of all } those bombarding electrons with nowhere to go), but then surely one } 15kV electron produces more than one secondary, doesn't it? } I've recently had an incident whereby I got EPMA analytical } totals which were a bit high (101 to 103), and this went away when I } was more liberal with the conductive paint. } Could this be because the sample was charging +ve, thus } increasing the effective accelerating voltage?
The only way the sample can charge up is if the BSE and SE current is more than the incident current. It is my understanding that for flat non-conducting samples that this situation occurs around 1kV, slightly less than the charge balancing condition for LVSEM. At higher voltages, the sum of the backscatter coefficient and secondary electron coefficients will be significantly less than one. If the sample is tilted to high angles (e.g., 70 deg), then the charge balance condition can be extended to about 3keV. But you wouldn't be tilting your sample to such high angles for analysis.
I think the answer to your increased signal may lie in the cross section for core ionization with overvoltage. If your sample charges negatively, then the overvoltage will decrease. The cross section for ionization increases as the overvoltage decreases. (It has a maximum at about 2.5-3.) Test this. When you have a good sample that is behaving properly, reduce the beam voltage a little and keep the probe current constant and see if the counts go up. - -Scott Walck
Micrion---Moving Beyond Expectations Put your talent to the test and job Micrion in the design, development, and manufacture of focused ion beam technology for semiconductor and disk drive industries. Micrion is recognized as a global leader in focused ion beam wafer modification and mask repair technology. As our business scope broadens to include magnetic head micromachining, we seek candidates who have an interest in leading edge technology and enjoy direct customer relations.
Applications Engineers
To provide technical marketing assistance on design and integration of customer applications relating to Focused Ion Beam systems, perform product demonstrations and technical presentations, research and develop customer applications and represent company at trade shows. BS technical degree, 2 years' related experience. Hands-on experience operating SEM, FIB, or process/analytical equipment desired. Strong communication skills, customer relations and a willingness to travel a must.
Micrion offers a competitive salary and benefits package, including but not limited to medical, dental, life insurance, tuition assistance, ESPP and 401K matching plan.
Reply to: Ms. Cheryl Cheney, Human Resources Manager
Mail: Micrion Corporation, 1 Corporation Way, Peabody, MA 01960-7990 Email: hr-at-micrion.com Web Site: www.micrion.com AA/EOE M/F/D/V
} } ...but will the drives be able to *write* (not just read) the current CD } } standard? If you want to distribute info widely (rather than just } } archiving), which Judy apparently wants to do, you don't want to do it on } } DVD until most people have access to drives that can read them. } } } } Alfred
I read an recent article about the development and marketing of DVD disks and drives. The introduction of writeable DVD drives is planned only for next year and of course it will take some time until they become affordable.
Petra -------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public Centre Universitaire (CRP-CU) Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpcu.lu or 100112.2335-at-compuserve.com
I, too, would be very interested in any first hand information or references available regarding the decalcification of bone - EDTA, ascorbic acid, or other.
Pat Hales Dept. of Anatomy & Cell Biology McGill University E-Mail: hales-at-hippo.medcor.mcgill.ca
Hi, there: One of my friends is going to buy a DVD writer. Does anybody know how much it costs and where to get information? Thanks in advance for your help. ------Weixin Xu------
A word to the wise. If you want people on a list to read your messages- don't send them as attachments- at least unless they are simple ASCII text files (but even that does not work for everyone). Attachments, often being specific to software, must be translated to be readable [this I know is a simplification]. Most of us won't take the time (or won't have the correct software) to accomplish the translation.
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-bgsm.edu * **************************************************************
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
It is a bit more complex than you think. Apart from the "overall" situation the impinging electrons have momentum. they know electronsout of the surface layer, creating a positive volume and are deposited farther in. Truely immense field are thus created if the resistivity is high enough. Eventually breakdown occurs and the electrons trapped inside can actually blast out through the surface into the vacuum (ballistic electrons).
It is a can of worms and makes one realize that EPMA on insulators is an approximate activity at best. There was work on this in terms of looking at frozen biological specimens (ice being an insulator). If memory serves, the fellow involved was Ricke, in Germany about 20 years ago and he found that the electrons didn't penetrate anywhere near as far as they were supposed to because the internal field slowed them down. Note, this was on coated specimens!! They also didn't make as many x-rays as they should becuse much of their initial kinetic energy was still stored in the field of the "virtual capacitor" near the surface.
Jim Pawley
**************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 1235, Engineering Research Building, FAX 608-265-5315 1500 Engineering Dr., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU
} } I've recently had an incident whereby I got EPMA analytical } } totals which were a bit high (101 to 103), and this went away when I } } was more liberal with the conductive paint.
Scott Walck added,
} I think the answer to your increased signal may lie in the cross section for } core ionization with overvoltage. If your sample charges negatively, then the } overvoltage will decrease. The cross section for ionization increases as the } overvoltage decreases. (It has a maximum at about 2.5-3.) Test this. When you } have a good sample that is behaving properly, reduce the beam voltage a little } and keep the probe current constant and see if the counts go up.
My own working definition for charging is that there is an avalanche of secondary electrons emitted from the sample. The practical effects of charging are that the electrons are slowed down going into the sample, because the negative local charge at the sample surface produces a force against the electrons arriving from the gun. The electrons may also be deflected away from the nominal beam spot. In terms of what you think you are analyzing, the beam may be on an adjacent grain, or on a grain boundary, etc. The beam could easily be deflected out of the lateral depth of field for the WDS spectrometers (several tens to hundreds of microns), the result being a lower x-ray intensity and therefore a low analytical total due to spectrometer defocusing.
Three ways to check for sample charging. First, the high energy cutoff of the EDS spectrum (Duane-Hunt limit) should ramp right up to the accelerating voltage. If it is routinely below that acc voltage, the sample is charging, and if it is above the acc voltage, you are seeing some pulse pileup -- that is ok. Secondly, if you are in imaging mode and you switch from very low mag right up to high mag, you may see a fairly rapid shift of the image due to beam deflection. Thirdly, you may see the absorbed current value jump around. Finally, for really bad charging, you see the streaking of the secondary electron image due to the SE avalanche. Of these it is my experience that the Duane-Hunt limit is most sensitive, and shows charging when none of these other features are observed. For this reason, you should acquire an EDS spectrum with each analysis to monitor the cutoff, as well as to check for missed elements.
The effect of electron retardation is to lower the overvoltage. This will reduce the generated x-ray intensity in the sample relative to the standard (assuming that the standard is fully conductive). If you are operating at an accelerating voltage that is 1.5 times the excitation energy of your most energetic line, then the ionization cross section is pretty linear in that range, and a small decrease in overvoltage should not really affect the cross section. Thus, a decrease in overvoltage (and intensity) will give you a low total, not a high one. If you are operating at a voltage that is right down on the excitation energy, then you are in the strongly curved portion of the cross section curve, and you might see an increase in intensity. But you should not be operating in this voltage regime to begin with, because we do not know the exact nature of the ionization cross section function. The same comment applies to a situation where, for example, the instrument is operating at 15KV but, due to charging, the sample is seeing an accelerating voltage more like 7 KV, i.e. really bad charging. This is what you would be talking about to really screw up a silicate analysis where Fe is your highest energy line.
Anyway, I don't really see why you should be getting high totals. You may wish to look beyond the topic of charging, like spectrometer alignment and reproducibility, standards, etc.
Paul
+----------------------------------------------------+ | Paul K. Carpenter paulc-at-gps.caltech.edu | | Division Analytical Facility | | Geological and Planetary Sciences MC 100-23 | | California Institute of Technology | | Pasadena, CA 91125 | | 818-395-6126 (X-ray Lab) 818-568-0935 (Dept. FAX) | +----------------------------------------------------+
I have a Polaron E5400 Sputter Coaterthat needs new gaskets. The gaskets I need go around the top and bottom of the glass sleeve(?). The thing that goes around the specimens and the sputtering thing sits on. Whatever... My old gaskets are getting cracked and that's probably causing it to take longer to pump down.
Does anyone out there know of a vendor on the West coast of the USA where I could buy my gaskets from?
Not only that, but what type of gasket do I need? Cuna? Viton? Neoprene? All I know is that it's black, rubber looking and kinda L shaped.
Thanks ahead of time for all the information.
Paula = ) The gasketless wonder. Electron Microscope Lab UC Berkeley
Thanks to Amanda Wilson, Bruce Wagner, and David Brauer for recommendations and comments. Bruce has recommended (pyro?) antimonate to precipitate Ca in place, localizing it, and I assume followed by TEM and electron probe to confirm the presence of Ca. I have heard of this technique, especially as applied to plants which employ sequestering of excess Ca as oxalate crystals in specialized cells called idioblasts. But I have also heard that your specimen must have very high levels of Ca present in some form, such as Ca crystals, in order for this method to work. So I also would assume that this method won't work for Ca which is held on the ion exchange groups of the cell wall, or as non-crystalline co-ions in the vacuole? Anyway, I will read up on the pyroantimonate method and see if it will track Ca even when it isn't a precipitate. Also will look at Alizarin red and try to translate to botanicals. Thanks, Janet.
Janet K. Dye Ph. D. Graduate Student, Soils Land, Air, and Water Resources University of California Davis, California USA (916) 752-0199 (916) 668-4217 jkdye-at-ucdavis.edu
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I wonder if anyone can help with this one?
I have a colleague who would like to embed a single cell after he has stimulated it with a fine carbon electrode which has been inserted into the cell. His eventual aim is to cut a section through the carbon electrode to see its intracellular orientation. However, to do this, he must keep the cell, with inserted electrode, in place on the microscope stage as he processes and embeds it - including resin polymerization.
This is the question:
Is there an EM embedding resin available that can be polylmerized without heating?
Could the regular Spuur or Epon mixes be modified by adding more DMP-30, or would that make the blocks too brittle to section?
The resins that polymerize under UV illumination would not work because we have no way of excluding oxygen from the resin surface.
If there is no good answer to this I suppose the next question should be: "what happens to an expensive light microscope after heating to 60#161#C for 8hr?"
Thanks in advance.
Paul Webster Center for Cell Imaging Yale School of Medicine http://info.med.yale.edu/cellimg
} It is a can of worms and makes one realize that EPMA on insulators is an } approximate activity at best. There was work on this in terms of looking } at frozen biological specimens (ice being an insulator). If memory serves, } the fellow involved was Ricke, in Germany about 20 years ago and he found } that the electrons didn't penetrate anywhere near as far as they were } supposed to because the internal field slowed them down. Note, this was on } coated specimens!! They also didn't make as many x-rays as they should } becuse much of their initial kinetic energy was still stored in the field } of the "virtual capacitor" near the surface. } } Jim Pawley
Do you have the reference for this? Or someone? Thanks! Phil
&&& Illigitimi non carborundum &&&&&&&& Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217)244-3145 days (217)355-3145 evenings oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
: Ritchie Sims asked, : : } } I've recently had an incident whereby I got EPMA analytical : } } totals which were a bit high (101 to 103), and this went away when I : } } was more liberal with the conductive paint.
and Paul K. Carpenter responded, : : The electrons may also be : deflected away from the nominal beam spot. In terms of what you think you : are analyzing, the beam may be on an adjacent grain, or on a grain : boundary, etc. The beam could easily be deflected out of the lateral depth : of field for the WDS spectrometers (several tens to hundreds of microns), : the result being a lower x-ray intensity and therefore a low analytical : total due to spectrometer defocusing. : : Anyway, I don't really see why you should be getting high totals. You may : wish to look beyond the topic of charging, like spectrometer alignment and : reproducibility, standards, etc. :
Beam deflection because of charging might even give high totals. I have seen beam line scans across a homogeneous sample have a maxima away from the zero deflection point. This occurred even when the spectrometer was "peaked" at the zero deflection point. Presumably this indicates some kind of spectrometer misalignment, but it could give high analytical totals.
Carl
Carl Henderson Electron Microbeam Analysis Laboratory University of Michigan 2005 C.C. Little Bldg. Ann Arbor, MI 48109-1063 USA -------------------------------- (313) 936-1550 (voice) (313) 763-4690 (FAX) chender-at-umich.edu (e-mail) --------------------------------
On 30 Jan 1997, Paul Webster wrote: } } I have a colleague who would like to embed a single cell after he has stimulated } it with a fine carbon electrode which has been inserted into the cell. His } eventual aim is to cut a section through the carbon electrode to see its } intracellular orientation. However, to do this, he must keep the cell, with } inserted electrode, in place on the microscope stage as he processes and embeds } it - including resin polymerization. } } This is the question: } } Is there an EM embedding resin available that can be polylmerized without } heating? }
Paul:
I have had reasonable success with a resin called Epo-Fix, sold by Electron Microscopy Sciences.
Weixin wrote; } } Hi, there: } One of my friends is going to buy a DVD writer. Does anybody know how } much it costs and where to get information? } Thanks in advance for your help.
We, Pioneer has demonstrated a prototype DVD-R at Japan Electronics Show last October. You can get some information from following WWWs. (Some of them are in Japanese with some pictures. Try your exploration!)
Dear Paul, There are several embedding resins that can be polymerized without heating, but I don't know about their sectioning capabitities. I use them to embed and polish bulk samples for SEM. They are Epokwik from Buehler, a two part epoxy that hardens in 0.5 hour and a cheaper alternative, called Jet-Set, that is made in Burnaby, B.C. These do heat themselves up a bit as they harden, but if you keep the volume low it shouldn't be too bad.
You wrote: } Is there an EM embedding resin available that can be polylmerized without } heating? } Regards, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
At last a technical question that doesn't sound like an extract from the parts list or instruction manual - how refreshing!
I can't help, though, unless you would like to come out here where I have a spare gasket and know somewhere to find more of these.
Date sent: Thu, 30 Jan 1997 14:39:26 -0800 (PST) To: microscopy-at-Sparc5.Microscopy.Com
Hey Kids!
I have a Polaron E5400 Sputter Coaterthat needs new gaskets. The gaskets I need go around the top and bottom of the glass sleeve(?). The thing that goes around the specimens and the sputtering thing sits on. Whatever... My old gaskets are getting cracked and that's probably causing it to take longer to pump down.
Does anyone out there know of a vendor on the West coast of the USA where I could buy my gaskets from?
Not only that, but what type of gasket do I need? Cuna? Viton? Neoprene? All I know is that it's black, rubber looking and kinda L shaped.
Thanks ahead of time for all the information.
Paula = ) The gasketless wonder. Electron Microscope Lab UC Berkeley
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa eurc-at-giraffe.ru.ac.za - tel: +27 461 318168 - fax: +27 461 24377
A Postdoctoral Research Assistant is required to provide analytical support for Geology Research using the SEM-EDX system. There may also be the opportunity to contribute to the undergraduate teaching programme if appropriate. The post will be tenable for a period of 13 months from 1st April 1997 to 30th April 1998. The post will commence on a salary of =A315,516 (point 3 on Researcher 'B' Scale). Application forms may be obtained from, and should be returned to, the Personnel Department, Oxford Brookes University, Gipsy Lane Campus, Headington, Oxford, OX3 0BP. The closing date for applications is 18th February 1997. Further details can be obtained from Dr R A Strachan (01865-483609 or e-mail rastrachan-at-brookes.ac.uk).
---------- } From: Paul Webster {paul.webster-at-Yale.edu} } } I have a colleague who would like to embed a single cell after he has stimulated } it with a fine carbon electrode which has been inserted into the cell. His } eventual aim is to cut a section through the carbon electrode to see its } intracellular orientation. However, to do this, he must keep the cell, with } inserted electrode, in place on the microscope stage as he processes and embeds } it - including resin polymerization. } } This is the question: } } Is there an EM embedding resin available that can be polylmerized without } heating? } } Could the regular Spuur or Epon mixes be modified by adding more DMP-30, or } would that make the blocks too brittle to section? } } The resins that polymerize under UV illumination would not work because we have } no way of excluding oxygen from the resin surface. } } If there is no good answer to this I suppose the next question should be: "what } happens to an expensive light microscope after heating to 60#161#C for 8hr?" ************************************** LR White with accelerator added cures at room temperature and below. LR Gold would cure at nasty minus temperatures. White intense light will do the curing. It's easy to keep out oxygen. Just set up a nitrogen cylinder with a two stage regulator and a tube outlet. Insert into the tube a pasteur pipette. Adjust the flow to give about a bubble a second with the pipette held in water. A cylinder at that flow rate will last many weeks. Use a retort clamp to fix the pipette within a few mm of the resin on the microscope slide. I expect that the light from the microscope itself - especially with a blue filter in place would cure the resin. With luck the nitrogen flow would also help to keep the specimen cool. I delightful experimental set-up to play with! I must mention that P&S sells LR White. I hope that this posting is of some help and I do not expect to sell a 44 gallon drum of the stuff to Paul Webster or anybody who is embedding single cells on slides. Cheers Jim Darley
Probing & Structure (Microscopy Supplies & Accessories) PO Box 111, Thuringowa QLD 4817 Australia
Phone +61 77 740 370 Fax: +61 77 892 313 A great microscopy site: http://www.ultra.net.au/~pns/
Although when you see the price, you may feel the gaskets are made from gold rather than rubber, many Polaron parts are available from Energy Beam Sciences, Inc., Agawam MA. The last # I have is (800) 992-9037.
Good Luk! Woody
My other address... woody.white-at-worldnet.att.net http://www.geocities.com/capecanaveral/3722
With a lower incident beam potential, the analysis volume (scatter) is smaller/more shallow. As the center analysis volume moves toward the material surface, resultant x-ray adsorption decreases. For the same energy input to the specimen, this can change not only countrates, but the overall shape of the acquired spectrum. This degree of effect is also a function of (at least) both the sample composition (softer x-rays are affected more than those of higher energy) and the beginning incident beam potential.
Woody
The other addresses... woody.white-at-worldnet.att.net http://www.geocities.com/capecanaveral/3722
{snip} core ionization with overvoltage. If your sample charges negatively, then the overvoltage will decrease. The cross section for ionization increases as the overvoltage decreases. (It has a maximum at about 2.5-3.) Test this. When you have a good sample that is behaving properly, reduce the beam voltage a little and keep the probe current constant and see if the counts go up. - -Scott Walck
Message-Id: {1.5.4.32.19970131141149.0069d310-at-biotech.ufl.edu} X-Sender: gwe-at-biotech.ufl.edu X-Mailer: Windows Eudora Light Version 1.5.4 (32) Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
I believe that Unicryl (Biocryl) resin can be polymerized with UV in the presence of oxygen. It is sold by SPI. } } } } } } } } } } } } } } } } } } } } } } At 05:58 PM 1/30/97 -0500, you wrote: -----------------------------------------. } } } I wonder if anyone can help with this one? } } I have a colleague who would like to embed a single cell after he has stimulated } it with a fine carbon electrode which has been inserted into the cell. His } eventual aim is to cut a section through the carbon electrode to see its } intracellular orientation. However, to do this, he must keep the cell, with } inserted electrode, in place on the microscope stage as he processes and embeds } it - including resin polymerization. } } This is the question: } } Is there an EM embedding resin available that can be polylmerized without } heating? } } Could the regular Spuur or Epon mixes be modified by adding more DMP-30, or } would that make the blocks too brittle to section? } } The resins that polymerize under UV illumination would not work because we have } no way of excluding oxygen from the resin surface. } } If there is no good answer to this I suppose the next question should be: "what } happens to an expensive light microscope after heating to 60#161#C for 8hr?" } } Thanks in advance. } } Paul Webster } Center for Cell Imaging } Yale School of Medicine } http://info.med.yale.edu/cellimg } } } ******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
In answer to Paul Webster's question about polymerization in situ on a microscope stage. We routinely polymerize LR Gold with a halogen headlamp on a ringstand. the lamp is connected to a 12 volt battery charger and works well. If you used this in conjunction with Jim Darley's advise on excluding oxygen, you should/may not to much of a problem. We surround the sample as much as possible with aluminum foil to concentrate the light from the lamp on the specimen. Hank Adams EML New Mexico State University
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello All,
I am trying to get some opinions/data in regards to the relative accuracy of quantitative EDS vs. WDS. The specific case is the analysis of nickel in gold plating cross sections. The plating contains approximately 2.5% nickel (to improve mechanical properties), with no other elements present. The samples are polished mounts. The plating is over 4 microns thick, with a nickel underplate of 3 microns.
We stay away from the underplate region and with this thickness, I don't think there should be any interaction volume size problems.
We are using quantitative EDS with standards. The standards are plating coupons that have been analyzed by AA to have approximately 2.2 and 2.5% nickel. The beam current, mags, spot size etc are kept the same for each sample and analyze both standards before and after the sample. On the sample, we analyze 3 separate areas (500 sec acquires) and average the results.
Our results seem to be very good for this technique. On the standards, we are always within 0 to 0.2 percent of the standard (ie: 2.22% standard is between 2.1 to 2.3 %). On the samples, if we get a reading which is more than 0.2% different than the group, we will throw that data point out and acquire another for the average. If the post analysis standards do not get within the 0.2% tolerance, we will throw out all three data points and re-run the entire test.
For this particular application, will WDS provide better results, and if so, how much better? Also, why would it be better in this case where there is no deconvolution necessary.
With pure element standards, the results were not as good. It seems intuitive that this would be true, is there any reason why pure standards would give better results when you have standards that bracket the composition?
Thanks for any advice or data,
John Giles Senior Materials Engineer Honeywell Space Systems
I know it's hard to keep up with all of the mergers, new company names and so on in this field. The former Polaron (later, BioRad) range of specimen preparation equipment is now manufactured by VG Microtech, in England, again under the "Polaron" trade name.
For those of you in the United States, Energy Beam Sciences has been appointed the exclusive agent for VG Microtech, and we provide spare parts (like gaskets) and consumables (like targets), along with bench service, for this equipment, even obsolete models. For those of you in Canada, the same service is provided by Soquelec.
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
International Symposium on Geology and the Environment Istanbul, Turkey September 1 to 5, 1997
There will be a session on "Materials and Biomedical Applications of Laser Scanning and Tandem Scanning Reflected Light Confocal Microscopy" as part of the above mentioned symposium. The confocal session is scheduled for Thursday, September 4. The symposiom has been organized in celebration of the 50th anniversary of the Geological Congress of Turkey.
The confocal session is being organized by Kenneth C. Moore of the University of Iowa. For additional information on this session he can be contacted at kenneth-moore-at-uiowa.edu. General information on the symposium can be found at
Thomas Moninger (moninger-at-emiris.iaf.uiowa.edu) Central Microscopy Research Facility http://www.uiowa.edu/~cemrf 85 EMRB University of Iowa Iowa City, IA 52242 319-335-8142 FAX 319-335-8049
Is anyone in this group familiar with the philosophical debates about whether the things one sees in a microscope should be regarded as "real" (realism) or simply "useful" (instrumentalism)? Prominent philosophers of science interested in this are Ian Hacking, Bas van Fraassen, etc.
I would like to hear from anyone who has any interest in this area, whether they have heard of the debate before or not. It may some day play a role in teaching.
Alfred
----------------------------------------- Alfred Kracher Geological Sciences Iowa State University Ames, IA 50011-3212 akracher-at-iastate.edu http://www.public.iastate.edu/~akracher vox:515 294 5439 fax:515 294 6049 -----------------------------------------
I've been able to embed isolated cells in LR White between glass slides then polymerize with UV. I keep the slides together using those black spring clamps. To keep the cells from getting squashed I usually Super-glue some coverslips on the ends of one of the slides. The slides need to be treated with a mold-parting compound first so you can get them apart later. The LRW polymerizes well except for the outer edges.
I can provide more details if needed.
--Page Owen Connecticut College Dept. of Botany New London, CT
Dear Paul, Another choice for an embedding resin might be "Unicryl" - made by British BioCell (no - I have absolutely no connection). Although I haven't yet had experience with it I have heard good reports about it with regard to tissue morphology and antigenicity at the EM level. According to the booklet of information about it, it will polymerize anywhere between -50C and 60C - either by heat or UV. They maintain that at lower temperature (4C to -20C) evaporation is minimal and the blocks may not need to be covered. One of my colleagues reports that at -10C the blocks polymerized under UV while still exposed to the air (no cover at all).
Pat Hales Dept. of Anatomy & Cell Biology McGill University
} I've recently had an incident whereby I got EPMA analytical } totals which were a bit high (101 to 103), and this went away when I } was more liberal with the conductive paint.
A possible cause may be the reduced absorption of X-rays in your sample.
The negative charging inside your sample will reduce the penetration depth of the electrons, thus increasing the amount and the energy of SE and BSE electrons, and thus reducing the amount of electron-energy available for X-ray production. So in combination with the lower effective penetration voltage percentages of less than 100 are usually expected.
But if your sample contains elements that emit X-rays that are heavily absorbed in the compound (esp. ultra-light elements) then the reduction of the amount of X-rays generated could be more than compensated by the fact that those X-ray that are still produced get to the surface much more easily, increasing the emitted X-ray intensity.
For further reading: Please see the article from Bastin and Heijligers in "Electron Probe Quantitation" (Ed. Heinrich & Newbury, Plenum 1991, pages 163-175), in which the EPMA of non-conductive specimens is discussed.
Hans Dijkstra
EDAX International European support office Tilburg, the Netherlands hans.dijkstra-at-edax.nl
} } It is a can of worms and makes one realize that EPMA on insulators is an } } approximate activity at best. There was work on this in terms of looking } } at frozen biological specimens (ice being an insulator). If memory serves, } } the fellow involved was Ricke, in Germany about 20 years ago and he found } } that the electrons didn't penetrate anywhere near as far as they were } } supposed to because the internal field slowed them down. Note, this was on } } coated specimens!! They also didn't make as many x-rays as they should } } becuse much of their initial kinetic energy was still stored in the field } } of the "virtual capacitor" near the surface. } } } } Jim Pawley } } Do you have the reference for this? Or someone? Thanks! } Phil
Here you go Phil and others.
Nothing on the Gernam group but the name Dorge comes to mind. Otherwise it is:
Pawley, J.B. (1972) Charging artifacts in the scanning electron microscope. Scanning Electron Microsc. 1972 (I), 153-160
Shaffner, T.H., Hearle, J.W.S. (1976) Recent advances in understanding specimen charging. Scanning Electron Microsc. 1976 (I), 61-70
**************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 1235, Engineering Research Building, FAX 608-265-5315 1500 Engineering Dr., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU
} Bruce has recommended (pyro?) antimonate to precipitate Ca in place,
Yes, pyroantimonate. This can, however, wreck havoc with the ultrastructure; try it and see.
} localizing it, and I assume followed by TEM and electron probe to confirm } the presence of Ca.
You can also locate the antimony by EDS.
} I have heard of this technique, especially as applied } to plants which employ sequestering of excess Ca as oxalate crystals in } specialized cells called idioblasts.
If you have ~ micron-sized CaC2O4 xtals, you can use EDS directly without using pyroantimonate--there is plenty of Ca in them.
} But I have also heard that your } specimen must have very high levels of Ca present in some form, such as Ca } crystals, in order for this method to work. So I also would assume that } this method won't work for Ca which is held on the ion exchange groups of } the cell wall, or as non-crystalline co-ions in the vacuole?
EDS needs a large fraction of a %, or ~mM concentrations (wet) to detect and quantitate an element. It is irrelevant what the chemical form of the element is. These amounts must only be present in the analysed volume; the overall amount can be very much less as long as it is present in small, concentrated regions.
} Anyway, I will read up on the pyroantimonate method and see if it will } track Ca even when it isn't a precipitate.
Any Ca++, and other Ca which has a greater affinity for pyroan- timonate than for its ligands will be precipitated. Good luck. Yours, Bill Tivol
oThe displacement of the idea that facts and evidence matter by the idea that everything boils down to subjective interests and perspectives is -- second only to American political campaigns -- the most prominent and pernicious manifestation of anti-intellectualism in our time.
-- Larry Laudan, Science and Relativism(1990), Quoted by Alan Sokal in article 2, below.
I refer those considering this question seriously to read the articles: 1. "Transgressing the Boundaries: Towards a Transformative Hermeneutics of Quantum Gravity " by Alan D. Sokal (published in Social Text)46/47, pp. 217-252 (spring/summer 1996).
-and the follow-up article:
2. "A Physicist Experiments With Cultural Studies"
also by Alan D. Sokal published in Lingua Franca, May/June 1996, pp. 62-64.
--------------------------------------
This may sound really bizarre to some of, but...
Is anyone in this group familiar with the philosophical debates about whether the things one sees in a microscope should be regarded as "real" (realism) or simply "useful" (instrumentalism)? Prominent philosophers of science interested in this are Ian Hacking, Bas van Fraassen, etc.
I would like to hear from anyone who has any interest in this area, whether they have heard of the debate before or not. It may some day play a role in teaching.
Alfred
----------------------------------------- Alfred Kracher Geological Sciences Iowa State University Ames, IA 50011-3212 akracher-at-iastate.edu http://www.public.iastate.edu/~akracher vox:515 294 5439 fax:515 294 6049 -----------------------------------------
------------------ RFC822 Header Follows ------------------ Received: by qmgate_backup.anl.gov with ADMIN;31 Jan 1997 14:52:18 U Received: by cmtnts3.cmt.anl.gov with SMTP (Microsoft Exchange Server Internet Mail Connector Version 4.0.994.63) id {01BC0F86.33B6BEE0-at-cmtnts3.cmt.anl.gov} ; Fri, 31 Jan 1997 14:51:12 -0600 Received: from sparc5.microscopy.com by cmtnts3.cmt.anl.gov with SMTP (Microsoft Exchange Internet Mail Connector Version 4.0.994.63) id CRMRZAVN; Fri, 31 Jan 1997 14:51:05 -0600 Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id KAA16034 for dist-Microscopy; Fri, 31 Jan 1997 10:24:46 -0600 Received: from mailhub.iastate.edu (mailhub.iastate.edu [129.186.1.102]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id KAA16031 for {microscopy-at-Sparc5.Microscopy.Com} ; Fri, 31 Jan 1997 10:24:44 -0600 Received: from [129.186.121.151] (kracher.geol.iastate.edu [129.186.121.151]) by mailhub.iastate.edu (8.7.3/8.7.3) with SMTP id KAA05379 for {microscopy-at-Sparc5.Microscopy.Com} ; Fri, 31 Jan 1997 10:32:35 -0600 (CST) Message-ID: {c=US%a=_%p=Argonne_National%l=CMTNTS39701312051CRMRZAVN-at-cmtnts3.cmt.anl.gov}
At 05:58 PM 1/30/97 -0500, you wrote: -----------------------------------------. } I wonder if anyone can help with this one? } } I have a colleague who would like to embed a single cell after he has } stimulated } it with a fine carbon electrode which has been inserted into the cell. His } eventual aim is to cut a section through the carbon electrode to see its } intracellular orientation. However, to do this, he must keep the cell, with } inserted electrode, in place on the microscope stage as he processes and } embeds } it - including resin polymerization. } } This is the question: } } Is there an EM embedding resin available that can be polylmerized without } heating?
Paul - You don't say what resolution your friend requires. Would it be possible to do this on the stage of a confocal scope, thus eliminating embedding & sectioning? As a group, the UV-polymerizing resins have miserable cutting characteristics, and I presume that the electrode is a lot harder than the cell. Sectioning this prep will not be easy.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117 Caspar, CA 95420
On Sat, 1 Feb 1997 22:50:32 -0600 (cst) Doug Keene {DRK-at-shcc.org} wrote:
} } Responding to a query of a few days ago, we routinely cut } bone and growth plate in this facility. Our processing } routine includes fixation in 1.5% glut / 1.5% } paraformaldehyde in 0.1 M cacodylate pH 7.4 containing } 6,000 ppm ruthenium hexamine trichloride (fix for 60 } minutes) followed by o.1m cacodylate with 6,000ppm RHT for } 15 minutes, followed by 1% OsO4 in the same buffer with } RHT. The buffer rinse after OsO4 should be 0.1M cacodylate } with no RHT. This is a somewhat modified } procedure of Hunziker et al., J Ultrastruct Res 8:1, 1984. } Dehydrate in 30, 50, 70, 90, 95 % EtOH, Propylene oxide, } infiltrate in Spurrs and polymerize at 70 C for 16 hours. } We face the block and section for LM using a glass knife, } then use a somewhat damaged area of our diamond for } ultrathin sections. We always have two diamond knives } open in the lab, one which is slowly getting trashed } cutting relatively soft tissue and one which is no longer } optimal and is now used to cut calcified samples. The } lifetime of the knives is usually about 7 to 8 months } before resharpening. Our blocks are often larger than 0.5 } x 1 mm. } } Hope this helps, } } Doug Keene } Shriners Hospital for Children } Portland (always raining) Oregon } ---------------------- } Doug Keene } DRK-at-shcc.org }
At 11:17 AM 1/31/97 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} For this particular application, will WDS provide better results, and if so, } how much better? Also, why would it be better in this case where there is no } deconvolution necessary.
John ...
It bothers me sometimes when EDX is compared with WDX, the accuracy for EDX is evaluated relative to having measured a known ... and claiming the result is "good", "excellent", or for that matter "close enuf" ...
My definitition of "quantitative analysis" also includes quantifying the error associated with the analysis. WDX makes this easy ... pure counting stats with regard to the integral and that associated with subtracting the background. It is the statistical evaluation of the background subtraction (let alone the error probagated by de-convoluting) which make evaluation of the counting error almost impossible with EDX.
It is true, that with today's computing power, the analyst wouldn't have to wait very long for an error associated with "modelling" the EDX continuum, but I don't see any EDX vendors delivering this error analysis. It is also true, the error associated with de-convolution, as common as it is needed, is also just as important. I'd certainly be interested in reading others' opinion on applying error analysis to the continuum, and subsequent determination of EDX sensitivities and detection limits.
So, my answer to your question, is there is no comparison ... if you want quantitation with accurate error analysis ... EDX is ^not^ your tool. However, I'm not saying it is inappropriate for Ni in Au at those compositions ... you don't appear to be near any detection limits. But, I would be careful with reporting your accuracies ...
cheers, shAf
{\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo ZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
Responding to a query of a few days ago, we routinely cut bone and growth plate in this facility. Our processing routine includes fixation in 1.5% glut / 1.5% paraformaldehyde in 0.1 M cacodylate pH 7.4 containing 6,000 ppm ruthenium hexamine trichloride (fix for 60 minutes) followed by o.1m cacodylate with 6,000ppm RHT for 15 minutes, followed by 1% OsO4 in the same buffer with RHT. The buffer rinse after OsO4 should be 0.1M cacodylate with no RHT. This is a somewhat modified procedure of Hunziker et al., J Ultrastruct Res 8:1, 1984. Dehydrate in 30, 50, 70, 90, 95 % EtOH, Propylene oxide, infiltrate in Spurrs and polymerize at 70 C for 16 hours. We face the block and section for LM using a glass knife, then use a somewhat damaged area of our diamond for ultrathin sections. We always have two diamond knives open in the lab, one which is slowly getting trashed cutting relatively soft tissue and one which is no longer optimal and is now used to cut calcified samples. The lifetime of the knives is usually about 7 to 8 months before resharpening. Our blocks are often larger than 0.5 x 1 mm.
Hope this helps,
Doug Keene Shriners Hospital for Children Portland (always raining) Oregon ---------------------- Doug Keene DRK-at-shcc.org
The detection limit of averaged (or, when feasible, high dose) Ca measurements with EPMA (=EDS) is about 0.3 mmole/kg dry wt.
On Fri, 31 Jan 1997, William Tivol wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear Janet, } } } Bruce has recommended (pyro?) antimonate to precipitate Ca in place, } } Yes, pyroantimonate. This can, however, wreck havoc with the } ultrastructure; try it and see. } } } localizing it, and I assume followed by TEM and electron probe to confirm } } the presence of Ca. } } You can also locate the antimony by EDS. } } } I have heard of this technique, especially as applied } } to plants which employ sequestering of excess Ca as oxalate crystals in } } specialized cells called idioblasts. } } If you have ~ micron-sized CaC2O4 xtals, you can use EDS directly } without using pyroantimonate--there is plenty of Ca in them. } } } But I have also heard that your } } specimen must have very high levels of Ca present in some form, such as Ca } } crystals, in order for this method to work. So I also would assume that } } this method won't work for Ca which is held on the ion exchange groups of } } the cell wall, or as non-crystalline co-ions in the vacuole? } } EDS needs a large fraction of a %, or ~mM concentrations (wet) to } detect and quantitate an element. It is irrelevant what the chemical form } of the element is. These amounts must only be present in the analysed } volume; the overall amount can be very much less as long as it is present } in small, concentrated regions. } } } Anyway, I will read up on the pyroantimonate method and see if it will } } track Ca even when it isn't a precipitate. } } Any Ca++, and other Ca which has a greater affinity for pyroan- } timonate than for its ligands will be precipitated. Good luck. } Yours, } Bill Tivol }
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
For anyone who is interested, there will be an EMBO Practical Course in Prague, Czech Republic this summer.
The course will focus on the application of immunocytochemical and stereological methods in biomedical research.
Prague 23 June - 2 July 1997
Electron Microscopy and Stereology in Molecular Cell Biology
Details can be found at the following URL:
http://info.med.yale.edu/cellimg/EMBO.html
Or from:
Paul Webster Center for Cell Imaging Yale School of Medicine http://info.med.yale.edu/cellimg Paul.Webster-at-Yale.edu Tel 203 785 3219 Fax 203 785 7226
For resin polymerization without heating, try LRGold resin. This resin polymerises with UV and at -10deg celsius. Be careful, as the distance between the UV light and your resin is very important for good polymerization. If you are interested, i will give you more information later.
Dev. Vaitilingon Free University of Brussels Marine Biology Lab. {dvaitili-at-ulb.ac.be}
Can anyone help with a Unicryl problem? I am using unicryl to fix root pieces for semi-thin sectioning. I am uv polymerising at -20, but the process is taking over 1wk no matter how close the Uv bulbs are to the beam capsules. I am using 6W Sylvania bulbs in an aluminium-foil lined box. The end result is often disappointing also with some samples requiring oven polymerisation to finish them off.
Any suggestions would be gratefully received.
Mark Munro The Soil biology unit SAC E-Mail m.munro-at-ab.sac.ac.uk
} Hello All, } } I am trying to get some opinions/data in regards to the relative accuracy of } quantitative EDS vs. WDS. The specific case is the analysis of nickel in gold } plating cross sections. The plating contains approximately 2.5% nickel (to } improve mechanical properties), with no other elements present. The samples are } polished mounts. The plating is over 4 microns thick, with a nickel underplate } of 3 microns.
snips
} For this particular application, will WDS provide better results, and if so, } how much better? Also, why would it be better in this case where there is no } deconvolution necessary. } } With pure element standards, the results were not as good. It seems intuitive } that this would be true, is there any reason why pure standards would give } better results when you have standards that bracket the composition? } } Thanks for any advice or data, } } John Giles } Senior Materials Engineer } Honeywell Space Systems
As has been mentioned, proper statistical analysis of the errors in EDX spectra (and WDX spectra, for that matter) is not undertaken in any commercial software packages, as far as I'm aware. However, the mechanisms of WDX mean that the user can themselves process the data more easily for proper error analysis.
WDX principally offers lower detection limits, by about an order of magnitude, and better energy resolution, which will improve your results if you are anlysing peaks which overlap in EDX.
I don't think either of these issues is relevant to your particular specimen.
You might find 'Quantitative electron-probe microanalysis' by Scott, Love and Reed, pub Ellis Horwood 1995, ISBN 0-13-104050-2 a useful reference.
Regards,
--------------------------------------------------------------- Dr L. P. Stoter Technical Editor, MICROSCOPY & ANALYSIS Technesis 17, Rocks Park Road email: LPS-at-teknesis.demon.co.uk Uckfield, E. Sussex Phone: +44 (0)1825 766911 TN22 2AT Fax: +44 (0)1825 766911 United Kingdom ---------------------------------------------------------------
I am a college senior who needs help finding resources on microscopy topics. I am doing a research paper on the applications of geology to the transmission/scanning electron microscope. I am particularly interested in soil analysis, fossils, or glacial geology. As of yet, I have had no luck in finding any sort of available resources. I need not only find resources, but narrow my topic. Could you recommend something or at least direct me to research done in this field? Thank you for your help and time.
Sincerely, Amy Linder at the University of Dubuque, Dubuque, IA
You should check the proceedings of the Annual Meetings of both the Microbeam Analysis Society and the Microscopy Society of America. I recall sessions on geological application over the last few years. It will not be alot but it will be a start...
Nestor Your Friendly Neighborhood SysOp
------cut---------- } in soil analysis, fossils, or glacial geology. As of yet, I have had no } luck in finding any sort of available resources. I need not only find } resources, but narrow my topic. Could you recommend something or at } least direct me to research done in this field?
} Amy Linder at the University of Dubuque, Dubuque, IA
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Mark Munro writes: "Can anyone help with a Unicryl problem? I am using unicryl to fix root pieces for semi-thin sectioning. I am uv polymerising at -20, but the process is taking over 1wk no matter how close the Uv bulbs are to the beam capsules. I am using 6W Sylvania bulbs in an aluminium-foil lined box. The end result is often disappointing also with some samples requiring oven polymerisation to finish them off."
Often, poor results with UV polymerization can be traced back to the age of the illumination being used. If the bulbs are old, try the polymerization process with new bulbs.
Paul Webster Center for Cell Imaging Yale School of Medicine http://info.med.yale.edu/cellimg
} I am a college senior who needs help finding resources on microscopy } topics. I am doing a research paper on the applications of geology to the } transmission/scanning electron microscope. I am particularly interested } in soil analysis, fossils, or glacial geology. As of yet, I have had no } luck in finding any sort of available resources. I need not only find } resources, but narrow my topic. Could you recommend something or at } least direct me to research done in this field? Thank you for your help } and time. } } Sincerely, } Amy Linder at the University of Dubuque, Dubuque, IA
I haven't any sources to hand, but the U. library should have the books/journals necessary by interlibrary loan at least. Iowa State will have the relevant materials if you're up for the 3 hour drive. Maybe U of Iowa. I'd limit your topic, depending on your interest, to either micropaleontology or mineralogy. Scanning EM has been used extensively in studying microfossils--foramenifera, conodonts, nannoliths (nannoplankton, a calcareous algae) and so on. If I remember right, there is a journal called "Micropaleontology", there definitely are books by that title (try subject and title searchs in BIP). Mineralogy uses both SEM and transmission EM. SEM is particularly useful because of EDX--energy dispersive x-ray analysis--that allows identification and rough quantitation of elements in a rock. This allows mineral identification; or helps, anyway. It should be discussed in a text on mineralogy. SEM & TEM are used to examine mineral structure as well. This topic should also be easily available in journals--find one recent ref and go nuts with its bibliography. Also try looking in Current Contents: find a likely-looking article or three from the title, and chase from there. Sorry I don't have more specific info., but someone else will. Also, this is enough to get you started--it's how I usually start my literature chases. Philip Oshel oshel-at-ux1.cso.uiuc.edu
I am in need of acquiring the conical part of the final lens of a JSM 35(version C) scanning electron microscope. The cone in question is the part of the lens that hangs down in the chamber. We intend to develop a modification. Please email me with the information so that further negotiations can be carried out. I should welcome also any information on the composition (and commercial specification) of the metal from which the cone is made of. Of course, info on where this metal can be obtained will be invaluable.
Thank you in anticipation of your cooperation and help.
Yours sincerely,
Jitu Shah
Dr. J. S. Shah H. H. Wills Physics Laboratory, University of Bristol Royal Fort, Tyndall Avenue, Bristol BS8 1Tl. UK. email: jss-at-siva.bristol.ac.uk Tel: 44 117 9288719 Fax: 44 117 9255624
I have been asked by a colleague about freeware/shareware which may be used to track the movements of particles or bodies, captured from the light microscope, and viewed on a PC . What he wants is something that can detect direction, distance of travel and/or speed. I think the idea is that he already has the captured images but needs to measure the motion of one or two particles in each of a lot of images.
Apparently there was a piece of software, written for the BBC computer ( a pre IBM PC invention in the UK) which could do this sort of thing for star pictures many years ago.
Have you considered encapsulating the specimen in warmed agar (I don't know what the lowest temperature agars are), acrylamide gel or something similar and then embedding the whole block normally afterwards. I could have misunderstood but I am assuming that once the sample is immobilized and fixed temperature may not be so critical.
Malcolm Haswell University of Sunderland UK
----------
I wonder if anyone can help with this one?
I have a colleague who would like to embed a single cell after he has stimulated. it with a fine carbon electrode which has been inserted into the cell. His eventual aim is to cut a section through the carbon electrode to see its intracellular orientation. However, to do this, he must keep the cell, with inserted electrode, in place on the microscope stage as he processes and embeds it - including resin polymerization.
This is the question:
Is there an EM embedding resin available that can be polylmerized without heating?
Could the regular Spuur or Epon mixes be modified by adding more DMP-30, or would that make the blocks too brittle to section?
The resins that polymerize under UV illumination would not work because we have no way of excluding oxygen from the resin surface.
If there is no good answer to this I suppose the next question should be: "what happens to an expensive light microscope after heating to 60#161#C for 8hr?"
Thanks in advance.
Paul Webster Center for Cell Imaging Yale School of Medicine http://info.med.yale.edu/cellimg
Message-Id: {1.5.4.32.19970203130911.00677b38-at-pop.fast.net} X-Sender: goldmrkr-at-pop.fast.net (Unverified) X-Mailer: Windows Eudora Light Version 1.5.4 (32) Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
At 09:11 AM 1/31/97 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Paul/Greg
Received comments from John Chandler, British BioCell Int Ltd in Cardiff for whom we distribute, regarding UNICRYL, which they manufacture. He says:
Of course you could use UNICRYL at 4C with UV, but all acrylic resins are softer than epoxy resins (the latter have aromatic cross-linking structures). It would be quite difficult to cut carbon inside the cell in a relatively soft resin. The whole thing looks impossible anyway to try and polymerize on the microscope stage with heat. If UV light could be passed down the microscope at the right wavelength then polymerization could take place that way, and a cover slip may be used to prevent evaporation from an open tray or to exclude oxygen, if necessary.
CONCLUSIONS 1. UNICRYL can polymerize under UV in the presence of oxygen. 2. Evaporation is minimal at low temperatures if the specimen can be kept cold on the microscope stage. 3. UNICRYL may or may not be hard enough to allow sectioning with carbon in the cells, depending on the size of the carbon wire.
Good Luck -
Don Cox
******************************************************** Donald P. Cox, Ph.D., M.B.A. GOLDMARK BIOLOGICALS/D.P. COX ASSOCIATES 437 Lock Street, Phillipsburg, NJ 08865-2764 (908) 859-2631 - - (908) 859-2875-FAX E-Mail: goldmrkr-at-fast.net/goldmarker-at-aol.com Web Page: http://members.aol.com/goldmarker
~~~"Goldmarking is everlasting probing!"~~~ ********************************************************
Re: Pure element standards vs. "near match" standards. The correction coefficients are typically larger when using pure element standards compared to standards nearly matching the unknown. Larger corrections can result in larger errors. The degree of this effect will vary, depending on the elements involved.
I apologise if this message arrives at the list twice but the first one seems t have gone to the wrong address.
Malcolm Haswell ----------
I have been asked by a colleague about freeware/shareware which may be used to track the movements of particles or bodies, captured from the light microscope, and viewed on a PC . What he wants is something that can detect direction, distance of travel and/or speed. I think the idea is that he already has the captured images but needs to measure the motion of one or two particles in each of a lot of images.
Apparently there was a piece of software, written for the BBC computer ( a pre IBM PC invention in the UK) which could do this sort of thing for star pictures many years ago.
I have a copy of a preprint for an article by H. Hoch which was published in Staining Technology but I do not have the full reference information (i.e. no title, no pages, no volume number). The paper deals with staining ultrathin sections of fungi with barium permanganate.
Can anyone help me out so that I can give correct credit where credit is due?
Thanks
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513-529-5712 Fax: 513-529-4243 E-mail: edelmare-at-muohio.edu
I'm not familiar with the philosophical debate, but for what it is worth, I always teach my microanatomy students that they never see an actual object in the real world OR in the microscope. What they see is the light reflected, transmitted through, diffracted, refracted, or emitted from an object. The IMAGES on their retinas are "real" because the light exists, but the images are only an approximation of the object, not the object itself.
Gary Radice, Associate Professor gradice-at-richmond.edu Department of Biology 804-289-8107 (voice) University of Richmond VA 23173 804-289-8233 (FAX)
Greetings, I agree that attacts on the rationalist stance of science are growing in frequency are important to rebut. My view of the error that the social constructionists make comes down to this. Consider the color red. When you look at a red object your brain "contructs" a color for that object. There can never be a way to know that the color that your brain paints that object in your mind is the very same color that my brain picks. Color in that sense is a "construct" of the mind. BUT, we can agree that the object in question is the SAME color, and agree that it corresponds to some reference color, obtained for example with a monochrometer. The constructionists argue because there are constucts in the mind that EVERYTHING in the mind is a construct. This can be easily disproved by the fact that we all agree about what color stop signs are.
Our agreement about the color of stop signs means that we can make another object, color it like a stop sign, and every one will agree about that one too. This sounds trivial, but it is at the core of our assurance about the reality of science.
Many people who doubt the reality of objects down the scope have never looked through a microscope. Perhaps the best thing we can do for such sceptics is to invite them into our labs and show then a rotifer, ascorbate crystals in polarized light, or a fly in SEM?
The geological journals are full of studies applying SEM/TEM/EPMA, among other microscopic techniques, to geologic problems. Look up the journals "American Mineralogist", "Journal of Sedimentary Petrology", and any journal on Paleontology/Micropaleontology (I don't know their titles). Also two general references that should point you in the direction are: "Electron Microscopy in Mineralogy" , Springer-Verlag, 1976, H.-R. Wenk, ed. And "Transmission electron microscopy of minerals and rocks", by Alex C. McLaren, Cambridge Univ. Press, 1991.
*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.* James J. McGee (jmcgee-at-sc.edu) Dept. of Geological Sciences University of South Carolina (803) 777-6300 (Office) Columbia, SC 29208 (803) 777-6610 (Fax)
----------
My bias is that reality exists if only by my own definition (the conclusions one jumps to may be only your own) however it is clear to me that by almost all rigorous recent (after 1940 or so) philosophic or "scientific" (misused here for emphasis) definitons, reality is unobservable. The most obvious example is the heisenberg uncertainty principle which states that one cannot observe both position and velocity of any moviing particle. Certainly looking to the night sky for stars is looking at ancient history. So to is looking at light images thru a microscope "ancient history" albeit on a smaller time scale. Point Number two: (are you counting??) My colleagues consider when asked that everything one observes thru a microscope to be "artifact"...but by golly I like my particular reliable artifact (some color stain, phase contrast,SEM,TEM etc). This is an important point when the uninitiated ask you if what they are seeing is "artifact".
jkdye-at-ucdavis.edu (J. K. Dye) wrote on the subject: Localizing Ca in Botanicals.
} Bruce Wagner has recommended (pyro?) antimonate to precipitate Ca in place, } localizing it, and I assume followed by TEM and electron probe to confirm } the presence of Ca. I have heard of this technique, especially as applied } specialized cells called idioblasts. But I have also heard that your } specimen must have very high levels of Ca present in some form, such as Ca } this method won't work for Ca which is held on the ion exchange groups of } Anyway, I will read up on the pyroantimonate method and see if it will } track Ca even when it isn't a precipitate. Also will look at Alizarin red } and try to translate to botanicals. } Thanks, Janet.
} Janet K. Dye Ph. D. Graduate Student, Soils Land, Air, and Water Resources University of California Davis, California USA (916) 752-0199 (916) 668-4217 jkdye-at-ucdavis.edu
Janet,
Charley Smalls and I used the pyroantinomate method years ago to localize Ca2+ in developing muscle at sites where Ca2+ is not normally crystaline. However, it also precipitates Mg2+ if I recall, so you have to do some special controls. The refs are in our paper: Smalls, C.M. & Goode,D. (1977) Ca+2 - accumulating components in developing skeletal muscle. J. Morphol.151: 213-238.
-Dennis
Dr. M. Dennis Goode Phone (301) 405-6917 Department of Zoology Fax (301) 314-9358 University of Maryland e-mail goode-at-zool.umd.edu College Park MD 20742 ************************************************************* "If the Lord Almighty had consulted me before embarking upon the creation, I should have recommended something simpler." -Alphonso X of Castile, 15th Century
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Reply to: Frontiers of Electron Microscopy in Materials Science...
April 20-25, 1998 7th Frontiers of Electron Microscopy in Materials Science Conference Irsee Germany Contact: W. E. King, L-356, LLNL, Livermore, CA 94551 E-mail: weking-at-llnl.gov
I am in need of acquiring the conical part of the final lens of a JSM 35(version C) scanning electron microscope. The cone in question is the part of the lens that hangs down in the chamber. We intend to develop a modification. Please email me with the information so that further negotiations can be carried out. I should welcome also any information on the composition (and commercial specification) of the metal from which the cone is made of. Of course, info on where this metal can be obtained will be invaluable.
Thank you in anticipation of your cooperation and help.
Yours sincerely,
Jitu Shah
Dr. J. S. Shah H. H. Wills Physics Laboratory, University of Bristol Royal Fort, Tyndall Avenue, Bristol BS8 1Tl. UK. email: jss-at-siva.bristol.ac.uk Tel: 44 117 9288719 Fax: 44 117 9255624
Dr.Jitu Shah H.H. Wills Physics Laboratory, University of Bristol, Royal Fort, Tyndall Avenue, Bristol BS8 1TL. UK email: jss-at-siva.bristol.ac.uk Tel: 44 117 9288719 Fax: 44 117 9255624
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I would appreciate the summary of confocal discussion that was posted sometime ago. I am interested on the properties of stand alone instruments since I already know and have the hardware to add digital confocal. 1) Basic stand alone price? 2) Options price 3) Gripes about instrument users have. 4) What made you decide on the instrument you have? 5) Realistic user/project ratio based on present utilization? 6) Does it take a full time percent effort to run the instrument? Thanks. ________ ___________ / ______/ / _________/ Cesar Danilo Fermin, Ph.D. / / / / Professor of Pathology & Otolaryngology / / / /_____ / \______ / ______/ Fax 504 587-7389 & Voice 584-2521 /________/ / / Internet: fermin-at-tmc.tulane.edu /_______ \/_/ | | | | http://www.tmc.tulane.edu/ferminlab | | | | | |______/ | |________ / Disclaimer: Whatever... is not Tulane's opinion!
I was wondering if anyone has tried using sodium borohydride (0.5 - 1.0 mg / ml ddH2O) on sections as a way to unmask antigens for immunolabelling. I have read that it is a good idea so I ordered some and discovered that sodium borohydride may not be a very safe chemical to have around.
Does anyone know of a technique either based on resin fixation or on a cryostage that can be used to image Phytophthora zoospores whilst keeping them intact?
Does anyone have available (especially in Australasia) a SiLi detector, complete with pre-amp and cryostat, and with reasonable resolution (~150eV)? If so, please let me know (incl. price) as we need to put one on our JEOL 35CF SEM (it can be suited to any SEM - we'll make our own vacuum interface).
Thanks,
Peter Smith, Dept. of App. Physics, RMIT, 124 LaTrobe St., Melbourne, Victoria, 3000, AUSTRALIA
After 17 years we have changed our business name from Probing & Structure to ProSciTech (caps are optional) and we have acquired a domain address for email and our on-line catalogue.
Nothing else has changed: management, ownership and or our commitment remain the same. The old internet addresses will continue to function, but please change your records and bookmarks to ProSciTech.
Regards Jim Darley
jim-at-proscitech.com.au
ProSciTech Microscopy Supplies & Accessories PO Box 111, Thuringowa QLD 4817 Australia
Phone +61 77 740 370 Fax: +61 77 892 313 A great microscopy site: http://www.proscitech.com.au
Bill Tivol, Dr. Somlyo, and Dennis Goode, Thank you for the advice. The detection limit for Ca by EPMA=EDS, or electron probe, is suprisingly high. My results with the fungus I work with would indicate a 100 x higher level of Ca held in exchangeable form on the cell walls, after exposure to realistic soil solution levels of Ca. And the literature on the host plant, Douglas Fir, would indicate Ca levels 25-30 x higher than the fungus (making some assumptions there). So, apparently, electron probe will do this, although I understand that asking a yes/no tracer question is a lot easier than asking how much Ca (and how exact?). Ca oxalate crystals have been observed to form in the walls and possibly in the vacuoles/vesicles of the structure I work with, so confirming that is not very interesting. What is interesting is where did the Ca come from is such quantities and where does it go, if anywhere. (The normal habitat for this symbiosis is an acidic, leached, relatively low Ca soil.) Maybe using Sr (stable) as a short term tracer for Ca is more useful. Still reading up and thinking, Janet.
Janet K. Dye Ph. D. Graduate Student, Soils Land, Air, and Water Resources University of California Davis, California USA (916) 752-0199 (916) 668-4217 jkdye-at-ucdavis.edu
} My bias is that reality exists if only by my own definition (the conclusions one } jumps to may be only your own) however it is clear to me that by almost all } rigorous recent (after 1940 or so) philosophic or "scientific" (misused here for } emphasis) definitons, reality is unobservable. The most obvious example is the } heisenberg uncertainty principle which states that one cannot observe both } position and velocity of any moviing particle.
I wouldn't paint such a black picture of quantum mechanics. QM is a theory of OBSERVABLES which means it ONLY makes statements about things that CAN be observed. The uncertainty principle states that e.g. location and momentum cannot be measured (or imposed on a particle) simultaneously with better than a certain precision. As I see it this has nothing to do with how well we can observe reality but is a property of reality itself. If You trap a particle in a potential (e.g. a quantum well) it will always have a certain kinetic energy (which is related to a momentum). If You make the potential well narrower the energy and momentum will increase. This is a direct consequence of the uncertainty principle and really the way a particle behaves. It is not a weakness of our observing powers.
Unfortunately QM is being misused a lot by people who want to prove that 'nothing is real' but it never said anything of the sort, just as Relativity never said that 'everthing is relative' and Goedel never said that 'every theory is either contradictory or incomplete'.
-- Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 93 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www_scem.csb.ki.se/pages/philip.html
I have been asked if there are any vendors of frame capture cards with some specific requirements - and I have come up blank and was hoping someone else be able to point us in the right direction.
RGB Input frame averaging
---} } and plugs into a PCMCIA laptop port. { {----
Or at least the ability to be utilized in a Laptop system.
Thanks.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513-529-5712 Fax: 513-529-4243 E-mail: edelmare-at-muohio.edu
Dear list, We are about to replace a 20 year old Technics Hummer VI sputter coater. Does anyone have opinions, strong or otherwise, as to who makes a descent one for ~$5000 or less (we don't need a Cr coater.) All individual responses will be kept as confidential as all the other government secrets.
Norman Elliott | Los Alamos National Laboratory MST-7 | PO Box 1663 MS E549 | Los Alamos, NM 87545
Philip Koeck wrote: {I wouldn't paint such a black picture of quantum mechanics. QM is a theory of OBSERVABLES which means it ONLY makes statements about things tha CAN be observed. The uncertainty principle states that e.g. location and momentum cannot be measured (or imposed on a particle) simultaneously with better than a certain precision.} Reply: Actually I wish to paint a full spectrum picture (color plus waves above and beyond "color") of quantum mechanics and indeed the changing face of the philosophy of science. To quote Shakespeare you have been "hoisted by your own pitard" (literally blown-up by your own bomb). It is not particularly useful in my opinion to invite philisophical discourse (especially in the future from students) and then totally limit their view of science and reality (or is that realities). You are quite correct in your view of science and reality from a 16th century point of view...indeed meterologists still talk about 20 yr "CYCLES" as if phenomena occured in round circles. "Science" changed forever about 1968 or so when it was realized that chaos theory indeed provides better models to explain natural phenomena than the 16th century "scientism" (which by the way correlates closely with strict religious philosophy of this era). For references I would strongly recommend author Ralph Abrahms esp titles about "Gaia, chaos, and eros"(not exact title). Biologists have been the last group to embrace this "new" (actually very old) way of looking at the universe..indeed they still misuse the phrases "good science" and "bad science" when sometimes all they are noting is the wild joyful ride of variation in all its "colors". PS In my informal poll of 10 oregonians all 10 thought reality(ies) "exist" but that they are not "observable" [substitute observable for measureable in the sentence you quote above] as we strictly understand. I suppose this could be attributed to the fact that it rains here a lot and soaks our brains.
This whole debate, which is more semantic than sensible, can be summed up in the painting of an apple by the surrealist Belgium painter Magritte which has a caption "ce ne pas une pomme"
Patrick Echlin Cambridge On Mon, 3 Feb 1997, Gary Radice wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I'm not familiar with the philosophical debate, but for what it is worth, I } always teach my microanatomy students that they never see an actual object } in the real world OR in the microscope. What they see is the light } reflected, transmitted through, diffracted, refracted, or emitted from an } object. The IMAGES on their retinas are "real" because the light exists, } but the images are only an approximation of the object, not the object } itself. } } Gary Radice, Associate Professor gradice-at-richmond.edu } Department of Biology 804-289-8107 (voice) } University of Richmond VA 23173 804-289-8233 (FAX) } } }
There's a very good book on sample preparation which may set your mind buzzing as to geology applications...
Section "Preparation of Rock, Mineral, Ceramic and Glassy Materials" from "Procedures in Electron Microscopy" Section 13.3 Ed,. AW Robards and AJ Wilson Publ. 1994
Good Luck
Tim Hazeldine ULTRA TEC MFG., INC. _________________________________________________________ *Manufacturing, Sales & Service 1025 E.Chestnut Avenue, Santa Ana, CA 92701-6491, USA Tel. 714 542 0608 Fax. 714 542 0627 Email. info-at-ultratecusa.com
*International Sales PO Box 312, Lincoln LN5 9XW, UK Tel./Fax. +44(0)1522 722833 Email. Ultratecex-at-aol.com __________________________________________________________
Precision Systems for Cutting, Lapping and polishing.......................... ___________________________________________________________
We have folks here who want to upgrade to a dependable cold cathode luminoscope (currently we have an unreliable hybrid setup)
Anyone out there know who makes the following cold cathode luminoscope: Citl CCL 8200 Mk3A (who is Citl?) (we've heard good things about it)
Similarly, we'd be interested in hearing about experiences with any recently purchased CCL systems. . Thanks.
John
John Fournelle Electron Microprobe Lab Internet:johnf-at-geology.wisc.edu Dept of Geology & Geophysics Office: (608) 262-7964 University of Wisconsin Lab: (608) 265-4798 1215 West Dayton Street Fax: (608) 262-0693 Madison, WI 53706 Amateur radio: WA3BTA/9 http://geology.wisc.edu/~johnf/sx51.html
"The first rule of all intelligent tinkering is to save every cog and wheel." Aldo Leopold
Sodium borohydride is indeed difficult to work with. A solution needs to be prepared fresh for each use, but if the container is left open the borohydride takes up moisture rapidly and will cake (no longer powder). When I used it years ago, I made up multiple 10 mg aliquots in capped microtubes, and stored them in a plastic container of silica gel dessicant in a -20 degree C freezer. When I needed a borohydride solution, I added a ml of ddH2O or buffer to an aliquot, and vortexed briefly (open the cap very quickly after vortexing, or the hydrogen gas evolving from the borohydride will pop it open, possibly causing a spill). Those aliquots have satisfied my needs since then.
A. Kent Christensen University of Michigan {akc-at-umich.edu}
--------------------------------------
On Mon, 3 Feb 1997, Beverly Phipps-Todd wrote:
} I was wondering if anyone has tried using sodium borohydride (0.5 - 1.0 mg } / ml ddH2O) on sections as a way to unmask antigens for immunolabelling. } I have read that it is a good idea so I ordered some and discovered that } sodium borohydride may not be a very safe chemical to have around. } } Bev. Phipps Todd } PhippTod-at-em.agr.ca }
Taking care not to be "hoisted by my own petard", I should like to carefully add a few comments on this subject.
Indeed Magritte did paint "Ceci n'est pas une pomme", which is what came to mind when I read the first message of the series.
Indeed what we look at are not cells but 2-dimensional representations of highly modified things that once were cells. (We have no doubt, I hope, that these cells are as round as the world).
However, the highly technical work which examined fully hydrated thin cryosection sections through vitrified biological tissues (eg McDowall et al 1983, J. Microsc. 131:1-9; 1984 J. Mol. Biol. 178:105-111; plus other work from the groups of Dubochet and Muller) do show that our pictures of chemically modified cells may well be on the way to being a representation of "the real thing" as they appear in 2-dimensions.
Our critical self doubt and constant search for new ways of imaging these structures may one day give us the ultimate - to image living processes inside living cells (in fact, this is possible for some intracellular processes).
Until then, we will have to make do with the brief moments in time captured in our 2-dimensional representations. For comfort we can hold onto the idea that collected facts predict unseen events? If we are able to examine our samples with a randomness that will give us a true representation of the whole population then we are lucky.
With this in mind, we now have to take care that we never "sell" the idea that cells consist of clearly defined organelles surrounded by white space all enclosed by two black lines. Not an easy task when the benchmark images are of highly extracted, high contrast images.
Keep Magritte in mind when presenting the micrographs:
"This is not a cell".
The implied message of course is:
"This is a picture of a cell"
Perhaps these comments are worthy only of smoking in Magritte's pipe ("Ceci n'est pas une pipe") but it will not stop my enjoyment of the discussion as it continues.
Regards,
Paul Webster, Ph.D. Center for Cell Imaging Yale School of Medicine http://info.med.yale.edu/cellimg
I have been trying to find some literature on the scanning electron microscopy of sugar cane stalk with particular reference to wax morphology. If anyone has encountered such literature or has personal experience I would appreciate some details. I am not a subscriber so will require a personal response. With thanks maryanne-at-one.com.au
Michael Goheen wrote: } Some folks on our campus have need of a Ralph Knife maker and have had no } success in finding one new or used. Any help would be greatly appreciated.
Energy Beam Sciences manufactures a Ralph Knife maker. We bought the rights to this product as part of the light microscopy specimen preparation product line formerly manufactured by BioRad in the U.K. Details are available on-line at our web site (http://www.ebsciences.com).
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
Here is something irresistable for all you Poe fans out there.
Abort, Retry, Ignore?
Once upon a midnight dreary, fingers cramped and vision bleary, System manuals piled high and wasted paper on the floor, Longing for the warmth of bed sheets, still I sat there doing spreadsheets. Having reached the bottom line I took a floppy from the drawer, I then invoked the SAVE command and waited for the disk to store, Only this and nothing more.
Deep into the monitor peering, long I sat there wond'ring, fearing, Doubting, while the disk kept churning, turning yet to churn some more. But the silence was unbroken, and the stillness gave no token. "Save!" I said, "You cursed mother! Save my data from before!" One thing did the phosphors answer, only this and nothing more, Just, "Abort, Retry, Ignore?"
Was this some occult illusion, some maniacal intrusion? These were choices undesired, ones I'd never faced before. Carefully I weighed the choices as the disk made impish noises. The cursor flashed, insistent, waiting, baiting me to type some more. Clearly I must press a key, choosing one and nothing more, From "Abort, Retry, Ignore?"
With fingers pale and trembling, slowly toward the keyboard bending, Longing for a happy ending, hoping all would be restored, Praying for some guarantee, timidly, I pressed a key. But on the screen there still persisted words appearing as before. Ghastly grim they blinked and taunted, haunted, as my patience wore, Saying "Abort, Retry, Ignore?"
I tried to catch the chips off guard, and pressed again, but twice as hard. I pleaded with the cursed machine: I begged and cried and then I swore. Now in mighty desperation, trying random combinations, Still there came the incantation, just as senseless as before. Cursor blinking, angrily winking, blinking nonsense as before. Reading, "Abort, Retry, Ignore?"
There I sat, distraught, exhausted, by my own machine accosted. Getting up I turned away and paced across the office floor. And then I saw a dreadful sight: a lightning bolt cut through the night. A gasp of horror overtook me, shook me to my very core. The lightning zapped my previous data, lost and gone forevermore. Not even, "Abort, Retry, Ignore?"
To this day I do not know the place to which lost data go. What demonic nether world us wrought where lost data will be stored, Beyond the reach of mortal souls, beyond the ether, into black holes? But sure as there's C, Pascal, Lotus, Ashton-Tate and more, You will be one day be left to wander, lost on some Plutonian shore, Pleading, "Abort, Retry, Ignore?"
My intention has been to ignore this thread entirely, but alas the enthusiasm of some responses has managed to reach out from the rows of words on my computer screen and cause my fingers to key this paragraph and the one below.
Life involves both replicable codes and excitations. Only the former can we put to paper or micrograph. However, the latter we can interact with. In the TEM, for example, we respond with hand on console to photons generated by electrons scattered by very tiny dynamical (albeit seldom living) objects in the path of the microscope's beam. Some signs of this interaction find their way to paper or disk, but never all of it since replicable codes (like pictures or spectra or words) hitch rides on only a miniscule subset of the dynamical excitations (ranging up in size from elementary particles through atoms to us) that we find in the world around. Hence the micrographs (and these words) are mere representations. They may or may not be of help. The interactions, however, are as real as are we.
These fellowships are intended to provide outstanding university faculty access to the SHaRE User Facility at Oak Ridge National Laboratory. This user facility is equipped with state-of-the-art electron microscopes, atom probe field ion microscopes, and mechanical properties microprobes for materials science microanalysis. These appointments are intended to assist faculty by enhancing their materials science research through extended access to SHaRE's state-of-the-art microanalytical facilities, and through collaborations with appropriate researchers at ORNL. It is anticipated that one junior and one senior university faculty will be appointed as fellows.
The duration of each fellowship is expected to be between six and twelve weeks. It is anticipated that fellowships will be taken during the participant's summer semester or quarter terms.
Eligibility Applicants must be full-time permanent faculty members at accredited U.S. colleges or universities.
Stipends and Allowances Stipends paid to participants are based on, but may not exceed, their regular college/university salary. The cost of travel for one round-trip between the academic institution and ORNL will be reimbursed if the distance is greater than 50 miles. Reimbursement is made according to the standard travel policy of the Oak Ridge Institute for Science and Education (ORISE).
Applications Application for fellowships may be made by submitting a written proposal, no more than four pages in length, to the below address. The proposal should:
=95Set forth the scientific or technological significance of the proposed research. =95State the relevance of research to the U.S. Department of Energy, and= ORNL. =95Include a statement of work which describes the major research tasks to= be performed, specifies needed research instrumentation, and indicates any anticipated specimen preparation at ORNL. =95Summarize the applicant's previous work in the field of the proposed research, including relevant expertise on the research equipment being proposed for use in this research. =95Identify a researcher at ORNL who agrees to collaborate in the research. SHaRE can assist faculty personnel in identifying appropriate ORNL collaborators. =95State the intended start date and duration of the appointment. =95Include a one-page professional resume.
Five copies of the proposal should be submitted to: SHaRE Faculty Fellowship Program Education and Training Division Oak Ridge Institute for Science and Education P.O. Box 117 Oak Ridge, TN 37831-0117
or a single copy submitted electronically to Ms. Renetta Godfrey (godfreyrd-at-ornl.gov). Supported file formats are ascii, and Word 6.0, and WordPerfect 7.0 (or earlier versions).
Review Process and Deadline =95Appointments are based on competitive evaluation of the applicants' qualifications, proposed plan of research, and relevance to ORNL/DOE research programs and activities =95Appointments are made by recommendation of the SHaRE Executive Committee =95Proposals for FY 1997 fellowships should be received by March 3, 1997
Additional Information For additional information regarding either SHaRE or these fellowships =95http://www.ms.ornl.gov/share/intro.htm =95contact Neal Evans evansnd-at-ornl.gov 423-576-4427
SHaRE Faculty Fellowships are contingent upon the availability of funds, collaborating personnel, and research facilities.
The Shared Research Equipment User Facility and Program are supported by the Division of Materials Sciences, U.S. Department of Energy, under contract DE-AC05-96OR22464 with Lockheed Martin Energy Research Corp., and through contract DE-AC05-76OR00033 with Oak Ridge Associated Universities.
The New England Society for Microscopy announces first meeting of 1997
PROGRAM WEDNESDAY, FEBRUARY 19, 1997
5 pm-----Registration and Tours of Philips Electroscan
6 pm-----Buffet Dinner
7:15 pm-----"Development of Recombinant Oral Vaccine Against Helicobacter pylori" to be presented by Thomas Ermak from OraVax, Inc.
8:00 pm-----"High Temperature Applications in Environmental SEM" to be presented by Thomas A. Hardt, Applications Development Manager at Philips Electroscan.
NEW MEMBERS WELCOME! Regular membership dues are $15 per calendar year. Registration for this meeting is $5.
To register, contact L. Kirstein at tel: 508-473-9673 or E-mail: 104365.3522-at-compuserve.com. (Mark message: NESM February Meeting Registration)
} I agree that attacts on the rationalist stance of science are } growing in frequency are important to rebut.
Not only that, but there is a long tradition of anti-intellectualism in America which must be counteracted. Otherwise, more rational people will see their economies expand while ours goes down the tubes.
} My view of the error that the } social constructionists make comes down to this. Consider the color red. } When you look at a red object your brain "contructs" a color for that } object. There can never be a way to know that the color that your brain } paints that object in your mind is the very same color that my brain picks.
If, as I think, a brain '"constructs" a color' by modifying synapses etc. which results in a particular neural firing pattern being associated with everything the brain's owner sees of that color, then everyone's brain constructs a different color, since the connectivity of neurons is almost certainly different for each individual brain. That said, however, there are likely to be similarities in the neural firing patterns which are con- structed, since each arises from signals from cones in the retina, which have features common to all individuals.
} Color in that sense is a "construct" of the mind. BUT, we can agree that } the object in question is the SAME color, and agree that it corresponds to } some reference color, obtained for example with a monochrometer. The } constructionists argue because there are constucts in the mind that } EVERYTHING in the mind is a construct. This can be easily disproved by the } fact that we all agree about what color stop signs are. } Not quite. The social constructionists would argue that your mind only constructs the agreement; i.e., I think that you and I agree about the color of stop signs, but you may think that we are agreeing about an entirely different construct.
} Our agreement about the color of stop signs means that we can make } another object, color it like a stop sign, and every one will agree about } that one too. This sounds trivial, but it is at the core of our assurance } about the reality of science. } That depends. You are only talking about people with normal color vision--an unstated assumption. You can easily make a second object which appears red, but whose spectrum of reflected or emitted light is quite dif- ferent from that of a stop sign. In any event, one can, however, specify a set of algorithms for the construction of an object and the measurement of some of its properties, and, if any number of other independent persons follow the same algorithms, they will all make the same observations. By this I mean that the observations will agree within some limits--it is very unlikely that any two measurements will yield *exactly* the same results. This whole subject gets quite complicated when all the details are consi- dered. My belief is that there is an actual reality which underlies each of our perceptions. No one's perceptions correspond exactly to that real- ity, nor even encompass more than an infinitessimal part of reality. By probing reality through making observations, recording our perceptions, correllating those perceptions which agree with those of others (including the magnitude and nature of expected disagreements), and rationalizing away those perceptions which do not agree, we can each and collectively set limits within which actual reality probably lies. I can assure any social constructionists that, if they perceive a boulder falling down a cliff heading directly toward them, they should act as though that boulder were actually real. Reality can always make an im- pact on any one of us whether or not we believe in it.
} Many people who doubt the reality of objects down the scope have } never looked through a microscope. Perhaps the best thing we can do for } such sceptics is to invite them into our labs and show then a rotifer, } ascorbate crystals in polarized light, or a fly in SEM? } These are, indeed, pretty constructs. What we can show sceptics are images of a rotifer (perhaps one which has been modified extensively), etc. What we see--and what they will see--are signals which have been manipulated so as to produce representations from which we can better understand the objects from which these representations arose. My lab does TEM of biological materials, and we rarely look at the biological material itself; we almost invariably look at the results of electrons scattered from a heavy metal stain. Fortunately, there are stains whose properties are well correllated to those of the biological specimen, so we can infer properties of the specimen from observations of the stain. I have no doubt that the biological specimen is real--as are the heavy metal atoms of the stain. I also have no doubt that what I see is only an approximation of some of the true properties of the system in which I am interested. Yours, Bill Tivol
AMRAY, Inc., a manufacturer of Scanning Electron Microscopes, invites you to browse our newly constructed WEB site. We can be found at www.amray.com. The WEB site includes information about AMRAY's 3000 series of scanning electron microscopes, our customer service organization, our customer training schools, and other imporatant information about AMRAY.
I would like to use a staining procedure (Masson's trichrome) that calls for picric acid for nuclear differentiation after staining with hematoxylin. Our safety officer would prefer it if I could avoid using picric acid. Is there a substitute? Is this step necessary?
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I would like to use a staining procedure (Masson's trichrome) that calls for picric acid for nuclear differentiation after staining with hematoxylin. Our safety officer would prefer it if I could avoid using picric acid. Is there a substitute? Is this step necessary?
My lab doesn't do any biological staining, but we do use picric acid solutions for metallographic etching. For years we went round and round with our safety people. They claimed it was not safe IF it was combined with SUFFICIENT amounts of certain metals (notably Pb) AND the subsequent picrate was subjected to a LARGE IMPACT, OR if the solution dried to below approximately 15% water AND the container was subjected to a LARGE SHOCK. Under the RIGHT circumstances it can be dangerous. So can a bottle of root beer.
The iron and steel industry has been using picric acid etchants for half century.
My personal opinion is that small (gram) quantities of wet picric acid, handled, stored, and disposed of properly by qualified personnel would not pose an extraordinary risk sufficient to prohibit its use.
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Diana,
My lab doesn't do any biological staining, but we do use picric acid solutions for metallographic etching. For years we went round and round with our safety people. They claimed it was not safe IF it was combined with SUFFICIENT amounts of certain metals (notably Pb) AND the subsequent picrate was subjected to a LARGE IMPACT, OR if the solution dried to below approximately 15% water AND the container was subjected to a LARGE SHOCK. Under the RIGHT circumstances it can be dangerous. So can a bottle of root beer.
The iron and steel industry has been using picric acid etchants for half century.
My personal opinion is that small (gram) quantities of wet picric acid, handled, stored, and disposed of properly by qualified personnel would not pose an extraordinary risk sufficient to prohibit its use.
Greetings, Many people seem to have liked the poem I sent round; but alas many have suggested that I wrote it. I have to make clear that I just forwarded it. I wish I had the time and skill to have written it. Sorry that wasn't clear in the first post.
i LIKE TO KNOW THE INFORMATION OF VENDORS PROVIDE THE LAMBS OF OLYMPUS BH SERIES OPTICAL MICROSCOPY.
BEST REGARDS,
Tseng-Ming Chou (Alex) Dept. of Materials Science and Engineering Stevens Institute of Technology Castle Point on Hudson, Hoboken, NJ 07030 e-mail: tchou-at-attila.stevens-tech.edu tchou-at-menger.eecs.stevens-tech.edu The Microstructure Group of Stevens
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Here is something irresistable for all you Poe fans out there.
Abort, Retry, Ignore?
Once upon a midnight dreary, fingers cramped and vision bleary, System manuals piled high and wasted paper on the floor, Longing for the warmth of bed sheets, still I sat there doing spreadsheets. Having reached the bottom line I took a floppy from the drawer, I then invoked the SAVE command and waited for the disk to store, Only this and nothing more.
Deep into the monitor peering, long I sat there wond'ring, fearing, Doubting, while the disk kept churning, turning yet to churn some more. But the silence was unbroken, and the stillness gave no token. "Save!" I said, "You cursed mother! Save my data from before!" One thing did the phosphors answer, only this and nothing more, Just, "Abort, Retry, Ignore?"
Was this some occult illusion, some maniacal intrusion? These were choices undesired, ones I'd never faced before. Carefully I weighed the choices as the disk made impish noises. The cursor flashed, insistent, waiting, baiting me to type some more. Clearly I must press a key, choosing one and nothing more, From "Abort, Retry, Ignore?"
With fingers pale and trembling, slowly toward the keyboard bending, Longing for a happy ending, hoping all would be restored, Praying for some guarantee, timidly, I pressed a key. But on the screen there still persisted words appearing as before. Ghastly grim they blinked and taunted, haunted, as my patience wore, Saying "Abort, Retry, Ignore?"
I tried to catch the chips off guard, and pressed again, but twice as hard. I pleaded with the cursed machine: I begged and cried and then I swore. Now in mighty desperation, trying random combinations, Still there came the incantation, just as senseless as before. Cursor blinking, angrily winking, blinking nonsense as before. Reading, "Abort, Retry, Ignore?"
There I sat, distraught, exhausted, by my own machine accosted. Getting up I turned away and paced across the office floor. And then I saw a dreadful sight: a lightning bolt cut through the night. A gasp of horror overtook me, shook me to my very core. The lightning zapped my previous data, lost and gone forevermore. Not even, "Abort, Retry, Ignore?"
To this day I do not know the place to which lost data go. What demonic nether world us wrought where lost data will be stored, Beyond the reach of mortal souls, beyond the ether, into black holes? But sure as there's C, Pascal, Lotus, Ashton-Tate and more, You will be one day be left to wander, lost on some Plutonian shore, Pleading, "Abort, Retry, Ignore?"
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Greetings, Many people seem to have liked the poem I sent round; but alas many have suggested that I wrote it. I have to make clear that I just forwarded it. I wish I had the time and skill to have written it. Sorry that wasn't clear in the first post.
I have had a request regarding suppliers of Utermohl-type sedimentation chambersfor standard quantitative hydrobiological analyses. My ignorance in this area is quite undiminished! All help gratefully received.
Best wishes
Keith Ryan
+++++++++++++++++++++++++++++++++++++++++++++++++Plymouth Marine Laboratory, Citadel Hill, Plymouth, Devon PL1 2PB, England
Can anyone of our subscribers reply to this person? Send the message direct to him as he is not on the Listserver.
Nestor
------------------
Below is the result of your feedback form. It was submitted by (Stankovic-at-fns.uniba.sk) on Thursday, February 6, 1997 at 05:40:07 ---------------------------------------------------------------------------
---------------------------- Forwarded with Changes ---------------------------
The TEM group at Intel Corp. in Santa Clara, CA announces a summer internship available for a graduate-level student in the area of focussed ion beam (FIB) milling. FIB milling is used in the semiconductor industry for specific-site thin sectioning of single-bit device failures for TEM analysis. The Ga+ ion beam typically used during FIB milling is known to amorphize the milled surfaces. The amorphous surface layers on the TEM section limit the scope, quality and utility of the TEM analysis. The goals of this internship are: 1.) to quantify the depth of amorphization imparted during FIB milling as a function of milling parameters (voltage, milling angle, etc.), 2.) to determine a low-damage milling condition and 3.) to develop a method to eliminate residual amorphization damage which remains after FIB milling. Prior experience with preparing and handling TEM samples is highly desirable. This position is for U.S. citizens or permanent residents, 3.0 gpa or higher, enrolled fulltime in school and able to work full time during the internship. Intel Corp. is an equal opportunity employer.
Please forward inquiries and resumes to:
David W. Susnitzky Intel Corporation 3065 Bowers Avenue, M.S. SC2-24 Santa Clara, CA 95052-8119
Destaining can be accomplished with 2% aqueous iron alum (ferric ammonium sulfate) instead of picric acid. Picric acid is said to give better definition of nuclei, though. I do not consider picric acid to be dangerous if stored "wet" (10% water) which is the way we get it from Fisher or whomever. Perhaps your Safety Officer could store the bottle for you and you could keep a saturated aqueous solution in the lab for use as needed.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (908)-235-4583; fax -4029 e-mail: mcauliff-at-.umdnj.edu ***************************************************************
On Wed, 5 Feb 1997, O=PRDSMTP; DDA.TYPE=RFC-822; DDA.VALUE=Microscopy-request(a)Spar wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Diana, } } My lab doesn't do any biological staining, but we do use picric acid } solutions for metallographic etching. For years we went round and round } with our safety people. They claimed it was not safe IF it was combined } with SUFFICIENT amounts of certain metals (notably Pb) AND the } subsequent picrate was subjected to a LARGE IMPACT, OR if the solution } dried to below approximately 15% water AND the container was subjected } to a LARGE SHOCK. Under the RIGHT circumstances it can be dangerous. } So can a bottle of root beer. } } The iron and steel industry has been using picric acid etchants for half } century. } } My personal opinion is that small (gram) quantities of wet picric acid, } handled, stored, and disposed of properly by qualified personnel would } not pose an extraordinary risk sufficient to prohibit its use. } } Harry Crossman } } Harry:
I thought I should reply after reading your message. About 15 years ago, I was working as an EM tech when a message came to the lab from the hazarads manager about a small vial of picric acid which had blown up in one of the labs on campus. It was a very old vial and appearantly had been forgotten. Noone was hurt, but I haven't forgotten the insident, and I make sure any excess picric acid is disoped of after the project requiring it is finished.
Any leads on how one might verify if two documents were written with the same pencil? Actually, the question is if something written in, say, 1948 can be distinguished from something written 10-20 years later (did the formulas for the pencil 'lead' change?). Perhaps pull off some lead particles with scotch tape and then examine with EDS/WDS? Any one know anything, or where to look?
Thanks.
John Fournelle Electron Microprobe Lab Internet:johnf-at-geology.wisc.edu Dept of Geology & Geophysics Office: (608) 262-7964 University of Wisconsin Lab: (608) 265-4798 1215 West Dayton Street Fax: (608) 262-0693 Madison, WI 53706 Amateur radio: WA3BTA/9 http://geology.wisc.edu/~johnf/sx51.html
"The first rule of all intelligent tinkering is to save every cog and wheel." Aldo Leopold
On Wed, 5 Feb 1997, O=PRDSMTP; DDA.TYPE=RFC-822; DDA.VALUE=Microscopy-request( a)Spar wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Diana, } } My lab doesn't do any biological staining, but we do use picric acid } solutions for metallographic etching. For years we went round and round } with our safety people. They claimed it was not safe IF it was combined } with SUFFICIENT amounts of certain metals (notably Pb) AND the } subsequent picrate was subjected to a LARGE IMPACT, OR if the solution } dried to below approximately 15% water AND the container was subjected } to a LARGE SHOCK. Under the RIGHT circumstances it can be dangerous. } So can a bottle of root beer. } } The iron and steel industry has been using picric acid etchants for half } century. } } My personal opinion is that small (gram) quantities of wet picric acid, } handled, stored, and disposed of properly by qualified personnel would } not pose an extraordinary risk sufficient to prohibit its use. } } Harry Crossman } } Harry:
I thought I should reply after reading your message. About 15 years ago, I was working as an EM tech when a message came to the lab from the hazarads manager about a small vial of picric acid which had blown up in one of the labs on campus. It was a very old vial and appearantly had been forgotten. Noone was hurt, but I haven't forgotten the insident, and I make sure any excess picric acid is disoped of after the project requiring it is finished.
The various comments that have appeared on the listserver concerning reality, philosophy, and science remind me of a story my high school teacher told to help distinguish the meaning of the words 'illusion', 'delusion', and 'hallucination'. It goes as follows:
Imagine a young person, who is somewhat superstitous and a bit afraid of the dark, walking alone along a deserted back road on a dark night. While passing an old abandoned house this person thinks he sees, out of the corner of his eye, a ghost in front of the house. Mustering all his courage, however, he stops to check the matter out. If on further investigation, 1. he finds nothing there, he suffered an illusion 2. he finds the remnants of a white sheet hanging from a clothsline, he had a delusion 3. he finds a ghost, he is having an hallucination
While this doesn't have anything to do directly with the questions at hand, I thought you might find it amusing
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
Perhaps someone can explain what I saw earlier today while examining a piece of archeological glass by SEM. When examining the sample at 10, 5 and 3 kV, I saw a distorted reflection of the secondary electron detector at the surface of the sample. The sample was attached to an aluminum stub using double-sided adhesive tape, and was not coated. The SEM is a Cambridge stereoscan 100. I did not observe any reflected image at higher kV.
The Alcoa Technical Center in Pittsburgh, PA announces a post-doc position for an electron microscopist.
The assignment is to characterize several aluminum alloy systems with regard to a) the nucleation mechanisms for recrystallization and the interaction of the growing grains with the microstructure, as well as b) the deformed structure (cell sizes, cell misorientation).
The following equipment is available: Philips 420 with TV camera, JEOL 840 with OIM attachment, FEG-SEMs at near-by universities, XRD, image processors. Besides an excellent background in the operation of TEMs and SEMs, a successful candidate should be familiar with EDS, foil thickness determination, analysis of Kikuchi patterns and quantitative stereology.
This position is limited to a period of two years. Alcoa is an equal opportunity employer.
Please forward inquiries and resumes to:
Hasso Weiland Alcoa Technical Center Alcoa Center, PA 15069
Final details of the 1997 Meeting of the Microscopical Society of Canada (Edmonton, 4 - 7 June) are now available. Lecture symposia and confirmed speakers include:
SCANNING-PROBE MICROSCOPY: Don Eigler, Paul Hansma, Richard Colton, Cynthia Goh, Darka Migus, Peter Grutter.
DYNAMICAL AND CONFOCAL MICROSCOPY: John White, Fred Fay, Lans Taylor, Winfried Denk.
ENERGY-FILTERED TEM: Peter Crozier, Richard Leapman, George Harauz, David Bazett-Jones.
SCANNING ELECTRON MICROSCOPY: David Joy, Arun Kumar, Arvid Lacis.
+ WORKSHOPS on Fundamentals of AFM, FEGSEM, EPMA and Confocal Microscopy.
2-page abstracts of contributed talks or posters are due 15 March. A registration package is being mailed to all MSC members; copies can also be obtained from:
Ray Egerton, Physics Dept, University of Alberta, Edmonton, Canada T6G 2J1 Phone: 403-492-5095, FAX: 403-492-0714, e-mail: egerton-at-phys.ualberta.ca
Further details of the meeting are available from the MSC Web Page: http://www.ualberta.ca/~mmid/msc ------------------------------------------------------------------------
I have an LKB 2178 Kinfemaker II that is having a problem. One of the C2 adjustment things no longer holds its' position. This results in the thing changing how it makes knives every single time you use it. I think that the C2 plastic parts are stretched out. It seems like it gets better if I take the top C2 apart and let it sit in the corner (by itself) for a day or two and then put it back together.
Has anyone else had this problem? Does anybody know where I can get the C2 plastic pieces so I can replace the worn out items?
One of my students tweaked all of the knobs one day and the knifemaker hasn't been the same since. Ah, the joys of a teaching lab!
Any suggestions are gratefully appreciated.
Desperately trying to make knives in Berkeley,
Paula = )
Paula Ssicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
This message is in MIME format. The first part should be readable text, while the remaining parts are likely unreadable without MIME-aware tools. Send mail to mime-at-docserver.cac.washington.edu for more info.
This is a forwarded message requested by Profs. Rodney C. Ewing and Lumin Wang at University of New Mexcio.
VACANCY ANNOUNCEMENT
TRANSMISSION ELECTRON MICROSCOPY ANALYTICAL ELECTRON MICROSCOPY -- LABORATORY MANAGER/RESEARCH SCIENTIST
Applications are invited for the position of laboratory manager/research scientist (Research Scientist III) supporting the transmission electron microscopy facilities in the Department of Earth and Planetary Sciences, University of New Mexico. The laboratory includes a JEOL 2010 high resolution TEM and JEOL 2000FX analytical STEM. Both instruments are equipped with EDS capabilities. More information on the lab may be obtained at HTTP//TEM.UNM.EDU.
We hope to fill this position by 1 May, 1997. The position is full time initially for 15 months and may be continued on a permanent basis. Duties will include supervision of all aspects of the electron microscopy laboratory, maintenance of the instruments, assistance to students, faculty, and research scientists at UNM, and outside users, and instruction of a graduate course in principles of electron-microscopy and use of the instruments. Time will be available for independent or collaborative research involving the use of the microscopes and other analytical facilities in the Department.
Candidates must hold a Ph.D. degree in earth science, materials science or a related field, and have a demonstrated research background in these disciplines, extensive skills in the operation and maintenance of transmission electron microscopes, and a record of published research activity. In addition, the candidate should have strong communication skills and an ability to work with and/or instruct individuals with broad research interests and backgrounds.
JOB TITLE: RESEARCH SCIENTIST III DEPARTMENT: EARTH AND PLANETARY SCIENCES REQUISITION NUMBER: 970231*A CLOSING DATE: 5:00 P.M. ON 3/21/97 GRADE 13
Based on Full-Time Salary: $2,858.42 to $3,801.42 mo. Full-time term fifteen month position with possibility of regular status.
IN ORDER TO BE QUALIFIED YOU MUST HAVE:
Ph.D. Degree in Technical, Scientific, or Engineering. Three (3) years experience directly related to the duties and responsibilities specified.
TO APPLY
Applications must be received by the Human Resources Office at 1717 Roma NE, or Health Sciences Ctr., Med Bldg. 2, Rm. 101, North Campus, Albuquerque, NM 87131 no later than 5:00 p.m. on the closing date, February 21, 1997. Resumes must list employment dates by month/year and must be accompanied by a cover letter. Functional resumes will not be accepted. Indicate the requisition number 970231*A and job title Research Scientist III on the application/cover letter. Application forms may be obtained by calling 505-277-6422.
I usually don't pay much attention to the numerous sample preparation discussions for biologic materials (I mostly deal with inorganics), so pardon me if this has been previously discussed.
I have been asked about the possibility of measuring elements in tree core using the electron probe. How best to prepare the wood for 10 - 20 kV, 20 nA , 30 sec. beam exposure? My only experience with wood usually involves an axe and a fireplace. Thanks in advance.
*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.* James J. McGee (jmcgee-at-sc.edu) Dept. of Geological Sciences University of South Carolina (803) 777-6300 (Office) Columbia, SC 29208 (803) 777-6610 (Fax)
To: Microscopy ListSer {Microscopy-at-MSA.Microscopy.Com}
James Martin wrote: } } Perhaps someone can explain what I saw earlier today while examining a } piece of archeological glass by SEM. When examining the sample at 10, 5 } and 3 kV, I saw a distorted reflection of the secondary electron detector } at the surface of the sample. The sample was attached to an aluminum stub } using double-sided adhesive tape, and was not coated. The SEM is a } Cambridge stereoscan 100. I did not observe any reflected image at higher } kV. } } Any thoughts? } } James Martin } Williamstown Art Conservation Center ................................................. James, You have encountered one of the neater artifacts you can accomplish with a SEM. What happened was that while imaging at 10 keV you established a fairly uniform charge on the surface of the glass. Then when you dropped down in energy, the incident electrons were elastically reflected by this uniform charge field and bounced backwards without ever hitting the specimen. In other words, the uniform charge field created a very nice "mirror" which reflected the electron beam towards your secondary detector so that's what you got an image of. It is also very typical to get a nice image of your final lens pole piece. The field created by this type of charging tends to be hemispherical in shape, so you get a kind of "fisheye" lens effect which at low mag allows you to get a "panoramic" image of the inside of your specimen chamber.
It is very easy to duplicate this phenomenon. I have found that a piece of smooth polysterene works quite nicely (I used a divider from a plastic parts bin). Simply charge the surface by scanning at low mag for a while at a high voltage and fairly high spot size and then switch to a lower keV and you will see the mirroring effect. A saphire bead also works very well. The better the insulator, the longer the effect lasts. You can actually get very nice images of the inards of your SEM. Take a few pictures and see if your microscopist friends can figure out how you did this!
This is really quite a fascinating effect which can really "blow your mind" if you stumble across it accidentally without knowing that such a thing is possible. This is such a neat effect that it just seems that there SHOULD be some good use for it -- Alas -- I don't know of any, other than to amuse yourself and your friends.
} Perhaps someone can explain what I saw earlier today while examining a } piece of archeological glass by SEM. When examining the sample at 10, 5 } and 3 kV, I saw a distorted reflection of the secondary electron detector } at the surface of the sample. The sample was attached to an aluminum stub } using double-sided adhesive tape, and was not coated. The SEM is a } Cambridge stereoscan 100. I did not observe any reflected image at higher } kV. } Some of the jolly service guys from Philips have performed a similar trick with uncoated styrofoam without knowing why it worked. They first bombarded the foam at 25 kV, then turned the acc. voltage down to 3kV or so. Here's my interpretation:
The glass, or styrofoam or whatever, charges up with electrons due to lack of grounding. As more primary electron bombard the sample they begin to be repelled by the like charge that has built up within the sample. When you use low kV the primary electrons are not able to penetrate the cloud of electrons around your sample, and are repelled by it. These electrons begin to hit the detector (what you saw), the final lens (what I saw with the styrofoam), or whatever else in the chamber, eliciting secondary electrons, and forming an image. What you see probably depends on the geometry of the chamber. We were able to look right up the final lens and see the aperture. At higher kVs you don't see the effect. Kinda cool, huh? Take a picture. Enjoy. Coat the sample and remove the coating after viewing.
I thunk this up myself - does this sound right?
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangela
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
I've seen the same when trying to image a minute marine snail that was poorly adfixed to the stub. I presumed that it was charging SO MUCH that the Primary Electrons were being completely repelled from the specimen back to the roof of the chamber. It looked great - a normal background with a shell-shaped "mirror" showing the ceiling of the chamber!
Geoff Avern Microscopy Labs Australian Museum Sydney, Australia
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Perhaps someone can explain what I saw earlier today while examining a piece of archeological glass by SEM. When examining the sample at 10, 5 and 3 kV, I saw a distorted reflection of the secondary electron detector at the surface of the sample. The sample was attached to an aluminum stub using double-sided adhesive tape, and was not coated. The SEM is a Cambridge stereoscan 100. I did not observe any reflected image at higher kV.
As you may know that the digital technology has came to join us in anywhere. It is no doubt about that the digital photography has a strong future in electron microscopy and image analysis. So I am going to purchase a digital camera and high quality laser print as well for our image analysis system. It may need to associate with among TEM, SEM and optical microscopy. Our TEM here is JEOL2000FX and SEM are JEOL 6400 and T300. But the trouble is that we only have got a limit budget which could be less than 10,000 pounds.
I am wondering if any body could give me more suggestion and information for them. Your advice would be very appreciation.
Thanks lot on advance.
Peiyi Wang Department of Engineering Materials University of Southampton Southampton SO17 1BJ UK Tel: 00441703 595101; Fax: 00441703 593016; E-mail: pw2-at-soton.ac.uk
I'm looking for information regarding the availability of colloidal carbon for use as an electron dense tracer in a vascular leakage model. I have found numerous references to Pelikan Ink, in particular Pelikan "Fount" Black India Ink No. 78. Pelikan still makes a No. 78 black ink, but it is not called Black India Ink. Does anyone know: 1) are these equivalent products, 2) are there other sources of colloidal carbon available for our studies, 3) has anyone tried using colloidal gold for these types of studies and would be willing to share their experience?
With regard to my third question, we are considering using BSA conjugated to 20 nm gold particles. At this point however, perfusion times, dilutions, etc. would all have to be empirically derived. I would greatly appreciate any insights from list members that might save us a lot of time and effort. Thanks in advance.
Fred Schamber and Tina Carvalho described a little gizmo which we sell. It's a lucite sphere mounted on carbon which will produce a reflected image. We call it a "Lumisphere".
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
I have been using a couple of diamond scribes sold by: Wale Apparatus Co, Inc. 400 Front Street Hellertown, PA 18055 phone 610-838-7047 FAX 610-838-7440
This is a glassworking and laboratory products company and their scribes work really well and come in several different flavors. They all have a pencil type handle and most have the diamond tip mounted on a 0.028"diameter metal shank. They are also reasonably priced.
I'm just a happy customer.
Hope this helps, Louie Kerr
} Subj: Need diamond scriber recommndations } } Can anybody recommend a good hand-held diamond scriber? } } I need something pretty robust but with small tip radius. I used to use a } scriber made by Fisher Scientific, but they stopped selling them. I mainly } scribe silicon wafers with varying amounts of metal and oxide layers. } } Becky Holdford } Texas Instruments / DMD Failure Analysis Lab } r-holdford-at-ti.com
Louie Kerr Research and Education Support Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543 508-289-7273 508-540-6902 (FAX)
I have two separate issues I'd like some input on. I'm somewhat constrained by how much info. I can provide so here's the bare bones.
1. I have a sample composed primarily of Fe and Cr. I'm characterizing the compositional Cr gradient as a function of location on the sample cross-section by EPMA. " Polished surface". My results for Cr concentration are nearly 20% lower than is expected based on analyses done in other labs by other techniques of which I have little or no knowledge of how it was done. I have reports that make compositional claims but give little or no methodology.
I'm using a 304 SS standard "nominal 18% Cr" for standardization. On my sample at a specific location where it is expected to measure 40 wt % Chromium, I'm getting in the range 30 to 35 Wt % Chromium.
As I said above this is the bare bones of the circumstance. Any input regarding Cr/Fe EPMA analysis will be appreciated.
ON ANOTHER FRONT
2. I'm toying with carbon analysis by EPMA. I've got a set of 6 carbon standards that with the exception of carbon concentration, are basically 52100 material. Their carbon ranges from 0.018 to 0.610 and they are very homogeneous. I'm using them to generate a curve from which I can use to pick off x-ray on peak count levels that relate to count rates from an unknown. I'm having trouble reconciling count differences between my curve generating standards and those of a SRM1225 which contains 0.275 carbon. All the standards have been verified by Spectrographic analysis. I've run as high as 300 points on the 1225 material and it consistently gives me lower average counts than would be expected based on the carbon curve.
Again any thoughts or suggestions will be appreciated " Thanks "
Terry R. McCue Babcock & Wilcox Research Metallurgical Analysis Section 1562 Beeson St. Alliance, Oh 44601 Phone: 330-829-7427 Internet: terry.r.mccue-at-mcdermott.com Fax: 330-829-7831
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
RE} detector reflected in SEM micrograph 2/7/97
I've seen a couple of responses attributing this effect to reflected electrons. Although I've never observed these "reflection" images (we religiously coat our glass samples, although I understand why one may not want to coat an art relic), I think I have a better explanation. The surface of the non-conducting sample is acting as part of a capacitor... the other part being the inside of the chamber. What you are imaging is a surface charge set up by this capacitance, and not reflected electrons. Electrons deflected completely away from the sample are unlikely to image much of anything (try running the EDS simultaneously with this effect...if the electrons are reflected there should be a huge bremstraalung peak, and nothing else.
--------------------------------------
Perhaps someone can explain what I saw earlier today while examining a piece of archeological glass by SEM. When examining the sample at 10, 5 and 3 kV, I saw a distorted reflection of the secondary electron detector at the surface of the sample. The sample was attached to an aluminum stub using double-sided adhesive tape, and was not coated. The SEM is a Cambridge stereoscan 100. I did not observe any reflected image at higher kV.
Any thoughts?
James Martin Williamstown Art Conservation Center
------------------ RFC822 Header Follows ------------------ Received: by qmgate_backup.anl.gov with ADMIN;6 Feb 1997 16:54:03 U Received: by cmtnts3.cmt.anl.gov with SMTP (Microsoft Exchange Server Internet Mail Connector Version 4.0.994.63) id {01BC144E.31278750-at-cmtnts3.cmt.anl.gov} ; Thu, 6 Feb 1997 16:52:52 -0600 Received: from sparc5.microscopy.com by cmtnts3.cmt.anl.gov with SMTP (Microsoft Exchange Internet Mail Connector Version 4.0.994.63) id DBN89C61; Thu, 6 Feb 1997 16:52:46 -0600 Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id MAA15575 for dist-Microscopy; Thu, 6 Feb 1997 12:26:32 -0600 Received: from williams.edu (goshen.williams.edu [137.165.4.3]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id MAA15572 for {microscopy-at-sparc5.microscopy.com} ; Thu, 6 Feb 1997 12:26:28 -0600 Received: from colrain.williams.edu (colrain.williams.edu [137.165.4.4]) by goshen.williams.edu (8.7.3/8.7.3) with SMTP id NAA27061 for {microscopy-at-sparc5.microscopy.com} ; Thu, 6 Feb 1997 13:34:39 -0500 (EST) Received: from localhost by colrain.williams.edu; (5.65v3.2/1.1.8.2/16Jul96-0543PM) id AA10258; Thu, 6 Feb 1997 13:34:39 -0500 Message-ID: {c=US%a=_%p=Argonne_National%l=CMTNTS39702062252DBN89C61-at-cmtnts3.cmt.anl.gov}
} Some of the jolly service guys from Philips have performed a similar trick } with uncoated styrofoam without knowing why it worked. They first } bombarded the foam at 25 kV, then turned the acc. voltage down to } 3kV or so. Here's my interpretation: } } The glass, or styrofoam or whatever, charges up with electrons due to lack } of grounding. As more primary electron bombard the sample they begin to } be repelled by the like charge that has built up within the sample. } When you use low kV the primary electrons are not able to penetrate } the cloud of electrons around your sample, and are repelled by it. These } electrons begin to hit the detector (what you saw), the final } lens (what I saw with the styrofoam), or whatever else in the chamber, } eliciting secondary electrons, and forming an image. What you see } probably depends on the geometry of the chamber. We were able to look } right up the final lens and see the aperture. At higher kVs you } don't see the effect. Kinda cool, huh? Take a picture. Enjoy. Coat the } sample and remove the coating after viewing. } } I thunk this up myself - does this sound right? } } Aloha, } Tina
Tina, Got it in one. Somebody (SPI? EDS?) used to sell a "specimen chamber inspection" stub that was basically half a marble that did this. Charged some outrageous price for it. Phil
P.S. I hope my "not the obvious" was taken as it was meant.
&&& Illigitimi non carborundum &&&&&&&& Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217)244-3145 days (217)355-3145 evenings oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
we need to replace an o-ring in our SEM to solve vacuum problems. If we order it, we'd have to buy the whole set of o-rings. According to our budget assigned for this year, this is not possible for us.
Our SEM is an old JEOL 35C. The o-ring we need is the one located at the bottom of the anode chamber. It's in viton and described by JEOL as G120, the dimensions are: internal diameter: 119,4 +- 0,4 mm thickness or width: 3,1 +- 0,1 mm
If anyone has extra ones and is willing to offer one to us, we would be happy to arrange something to get it (buying it, exchanging it for other part...)
We appreciate very much your attention. Thanks,
Silvia Montoro Centro Regional de Investigacion y Desarrollo de Santa Fe Santa Fe - Argentina csedax-at-arcride.edu.ar
we need to replace an o-ring in our SEM to solve vacuum problems. If we order it, we'd have to buy the whole set of o-rings. According to our budget assigned for this year, this is not possible for us.
Our SEM is an old JEOL 35C. The o-ring we need is the one located at the bottom of the anode chamber. It's in viton and described by JEOL as G120, the dimensions are: internal diameter: 119,4 +- 0,4 mm thickness or width: 3,1 +- 0,1 mm
If anyone has extra ones and is willing to offer one to us, we would be happy to arrange something to get it (buying it, exchanging it for other part...)
We appreciate very much your attention. Thanks,
Silvia Montoro Centro Regional de Investigacion y Desarrollo de Santa Fe Santa Fe - Argentina csedax-at-arcride.edu.ar
A colleague has had difficulty handling Ni grids because of the high degree of static electricity -- one of the side effects of the dry laboratory air during Feb. in the Great White North. He has shunned his woolen sweaters, has been using appropriate tweezers and when staining has beenn wetting the forceps to reduce the problem. The critical step is of course when inserting the grids into the holder for viewing in the TEM. Short of purchasing an anti-static device, are there any ingenious tips to overcome the case of the leaping grids? Thanks.
Carolyn J. Emerson email: cemerson-at-plato.ucs.mun.ca
Biology Department Memorial University St. John's, NF A1B 3X9 Tel: (709) 737-7515 Fax: (709) 737-3018
--IMA.Boundary.883533558 Content-Type: text/plain; charset=US-ASCII Content-Transfer-Encoding: 7bit Content-Description: cc:Mail note part
Fred et al. Actually, this effect does have one useful application: if you suspect something wrong inside the chamber, such as a loose wire, or a cold stage tubing gone amuck, you can check it out this way without having to open up the chamber to atmos. That's the only use I've found; any others?
Damian Neuberger Baxter International neuberd-at-baxter.com
James, You have encountered one of the neater artifacts you can accomplish with a SEM. ..... This is such a neat effect that it just seems that there SHOULD be some good use for it -- Alas -- I don't know of any, other than to amuse yourself and your friends.
Fred Schamber --IMA.Boundary.883533558 Content-Type: text/plain; charset=US-ASCII; name="RFC822 message headers" Content-Transfer-Encoding: 7bit Content-Description: cc:Mail note part Content-Disposition: inline; filename="RFC822 message headers"
Received: from mpg.mcgawpark.baxter.com (159.198.180.50) by ccmailgw.mcgawpark.baxter.com with SMTP (IMA Internet Exchange 2.1 Enterprise) id 0006E398; Fri, 7 Feb 97 00:52:43 -0600 Received: from sparc5.microscopy.com by mpg.mcgawpark.baxter.com id aa26142; 7 Feb 97 0:52 CST Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id VAA17473 for dist-Microscopy; Thu, 6 Feb 1997 21:07:21 -0600 Received: from orion.bv.sgi.net (orion.bv.sgi.net [206.150.193.66]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id VAA17468 for {Microscopy-at-Sparc5.Microscopy.Com} ; Thu, 6 Feb 1997 21:07:17 -0600 Received: from Stargate.sgi.net (dialup65.pgh.sgi.net [206.151.183.67]) by orion.bv.sgi.net (8.8.5/8.8.5) with SMTP id WAA24570 for {Microscopy-at-Sparc5.Microscopy.Com} ; Thu, 6 Feb 1997 22:14:59 -0500 (EST) Message-ID: {32FA9DE2.668D-at-sgi.net}
The authentication of documents is a big deal for the legal profession so try contacting your local/state bar association or look in classified ads in their newsletter/journal. There are companies that specialize in this sort of thing.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (908)-235-4583; fax -4029 e-mail: mcauliff-at-.umdnj.edu ***************************************************************
Are you sure it is static and not magnetic tweezers? Try non-magnetic ones. - -Scott
- ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
A colleague has had difficulty handling Ni grids because of the high degree of static electricity -- one of the side effects of the dry laboratory air during Feb. in the Great White North. He has shunned his woolen sweaters, has been using appropriate tweezers and when staining has beenn wetting the forceps to reduce the problem. The critical step is of course when inserting the grids into the holder for viewing in the TEM. Short of purchasing an anti-static device, are there any ingenious tips to overcome the case of the leaping grids? Thanks.
Carolyn J. Emerson email: cemerson-at-plato.ucs.mun.ca
Biology Department Memorial University St. John's, NF A1B 3X9 Tel: (709) 737-7515 Fax: (709) 737-3018
The authentication of documents is a very big deal in the legal profession and there are companies that specialize in this work. Try your local/state bar association or their journal/newsletter.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (908)-235-4583; fax -4029 e-mail: mcauliff-at-.umdnj.edu ***************************************************************
Hello fellow microscopists, Could anyone provide us with information on upgrading a Zeiss Axiovert 10 inverted fluorescence microscope with confocal capabilities? Are there relatively inexpensive kits available in setting up confocal or do you have to go the route of a manufacture's design? In addition are there any good software programs available for 3-D reconstruction of confocal Z series, etc.?
Thanks in advance. Dennis Kunkel
*********************************************** * Dennis Kunkel Ph.D. * * Pacific Biomedical Research Center * * University of Hawaii * * * * email - kunkel-at-pbrc.hawaii.edu * * www - http://www.pbrc.hawaii.edu/~kunkel/ * ***********************************************
Over the past few years I have been collecting the components of an EDS system for our TEM. I think I have all the parts, and now hope to put them together in the most efficient and cost effective manner (I also want it to work!).
Here is what I have:
Kevex detector to fit our microscope.
Kevex 7000 chassis with 4505P pulse processor, bias power supply is somewhere inside, no Kevex software, dead disk drive and rest of system appears to be dead too.
NIM bin with another 4505P and PGT bias power supply modules.
Link model 1134 bias power supply and PP, newer would like to use this if possible.
Dapple X-Mate MCA and acquisition controller.
A Link detector that does not fit our microscope.
Questions:
If I can figure out the plugs and sockets, could I use the Link PP and bias on the Kevex detector? I would like to do this because the Link box is newer and a better shape than the Kevex 4505P either in a NIM bin or in the old Kevex 7000 chassis.
What is the chance of finding out the pin outs and adjustments needed to mate the weird combination of plugs and pins I will end up with in a hybrid system?
How nuts do you think I am for to try to piece together a system this way as opposed to just getting all the components from a single source?
We are kind of Mac oriented in the lab, so I was thinking of a Mac MCA (maybe 4pi) board to collect and work over spectra once I get some of the detector/bias/PP part worked out.
What are some of the other pitfalls I should watch out for in putting something like this together? We might have as much as $10K to devote to this project, that would have to go for the new 4pi or other board and the modifications to the components.
Any ideas for a better plan? Anybody want the leftovers if it works?
Thanks for your patience.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
REDUCED FEE REGISTRATION DEADLINE April 30, 1997 Workshop on Tripod Polishing
Workshop Objective This course will cover all aspects of pre-thinning and focus on final thinning for TEM via Tripod Polishing. Due to the limited class size and the extensive hands-on opportuinities, this course is well suited to novices as well as advanced Tripodders. Attendees will also learn the latest techniques available in ion milling and in plasma cleaning for TEM samples. The course will include sections on:
How to do it and why should I? What's really going on and what am I really seeing? How to prepare small, specific area cross-sections. The problem of wildly differing materials (eg tungsten). Rapid preparation of TEM cross-sections. Preparation of a wide range of materials: semiconductors, ceramics, metals,...
Hands-on Opportunity This course will be unique in that it will provide a hands-on opportunity for every class participant. Tripod Polishers, Polishing Wheels, and pre-thinning equipment will be made available to all participants and actual samples will be prepared - by the students - as part of the course. This is a great opportunity to get your hands dirty and actually learn by doing. The instructors will walk you through each step of the process and then let you loose on the equipment. This course is designed to teach the Tripod Polishing technique. Silicon samples will be provided to the students and used as the basis for the course teaching.
Workshop Location and Dates South Bay Technology - San Clemente, CA Dates: Friday & Saturday - June 6 & 7, 1997
Previous Participants (partial list) INTEL, AMD, Motorola, LSI Logic, Conner Peripherals, Univ of Maryland, Univ of New Mexico, UNAM (Mexico), LG Electronics (Korea), Battelle, MEMC, MVA Inc., Univ of Michigan, U.S. Bureau of Mines, IBM, Naval Research Lab, Purdue Univ, Univ of Alabama, Univ of Arizona, Univ of Colorado, Univ of Wisconsin.
Class Size Due to the intensive hands-on aspects of this course, class size will be strictly limited to 10 participants.
Registration Fee: $795 (includes lunches and Friday night Dinner) $695 if registration fee paid by April 15, 1997
Registration Deadline: 30 days prior to workshop
For additional Information: Diane Macdonald South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 TEL: 800-728-2233 FAX: 714-492-1499 e-mail: sbt-at-southbaytech.com
ON-LINE Registration available at: http://www.southbaytech.com
Registration Form
To register for the workshop, please fill out this form and send it, with registration fee to:
South Bay Technology, Inc. Workshop on Tripod Polishing 1120 Via Callejon San Clemente, CA 92673 USA
Payment must be made in the form of a check, money order, Visa or MasterCard. Checks must be drawn on a U.S. Bank and made payable to South Bay Technology, Inc. Credit card orders by FAX may be sent to South Bay Technology at 714-492-1499. Please do not send credit card information via e-mail.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Subject: Time:4:58 PM OFFICE MEMO Job Opportunity at NCEM Date:2/7/97
Staff Scientist Opening
The Materials Sciences Division of the E. O. Lawrence Berkeley National Laboratory has an immediate opening for a full time Staff Scientist to work in the National Center for Electron Microscopy (NCEM).
NCEM is a national user facility with several state of the art electron microscopes and advanced image analysis and specimen preparation facilities. We are currently looking for an enthusiastic materials scientist to develop the In Situ Microscopy program at NCEM. The successful candidate will conduct original research in materials science utilizing advanced techniques of electron microscopy with a focus on mechanisms and dynamics of transformations and reactions at internal interfaces. The candidate will lead the development and operation of the In Situ facility which includes a 1.5MeV High Voltage Microscope, a 200keV In Situ microscope, and the facility's specimen preparation laboratory. The position offers a chance to explore a broad range of research opportunities by initiating collaborative projects with other internal and external investigators, conceiving novel experiments, developing new microscopy techniques, sample configurations or instrumentation. The candidate will contribute significantly to the future development of the facility.
The position requires a strong background in transmission electron microscopy and current practical experience in dynamic experimentation, specimen preparation and advanced microscopy techniques such as high resolution imaging, high voltage microscopy, convergent beam diffraction, microanalytical techniques, or computer image analysis/interpretation. An essential requirement is the ability to initiate collaborations and to carry out high quality research using the unique facilities of the NCEM. A Ph.D. in the physical sciences is highly desirable.
Please send resume and cover letter to Lawrence Berkeley National Laboratory, Staffing Office, Job #MSD/4891, One Cyclotron Road, MS938A, Berkeley, California 94720.
For more information, see Current Job Offers at --- http://www.lbl.gov/LBL-Documents/CJOs The job description is at -- http://www.lbl.gov/LBL-Documents/CJOs/sci4891msd.html
} A colleague has had difficulty handling Ni grids because of the } high degree of static electricity -- one of the side effects of } the dry laboratory air during Feb. in the Great White North. } He has shunned his woolen sweaters, has been using appropriate } tweezers and when staining has beenn wetting the forceps to } reduce the problem. The critical step is of course when inserting } the grids into the holder for viewing in the TEM. Short of } purchasing an anti-static device, are there any ingenious tips } to overcome the case of the leaping grids? Thanks. } } Carolyn J. Emerson
Dry air in St. John's?! A major climatic shift. A quick-and-dirty try: wrap a clean wire around the specimen holder (on the outside-of-the-vacuum side of the o-ring), and run the wire to anything grounded--a bit of bare metal on the EM's chassis, a water pipe, whatever's handy where you load the grids into the holder. Have 2 wires, one grounded with a free end--touch the forceps to the wire, grab the grid, touch wire to forceps again (being paranoid, like all good EM people), then load. Phil
&&& Illigitimi non carborundum &&&&&&&& Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217)244-3145 days (217)355-3145 evenings oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
I am in the process of attempting to make serial sections of a heterogeneous rock, and use algorithms in Mathematica to reconstruct 3-D structures. My question: Is someone familiar with a paper or book which can be used to determine the distance necessary between adjacent serial sections, given the size of structures that I am attempting to join up between sections, for a given statistical significance?
Any information would be greatly appreciated.
Phil Piccoli
***************************************************************************** Phil Piccoli Assistant Research Scientist Department of Geology Univ. of Maryland at College Park College Park, MD 20742-4211
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello, fellow microscopists!
Fred Schamber and Tina Carvalho described a little gizmo which we sell. It's a lucite sphere mounted on carbon which will produce a reflected image. We call it a "Lumisphere".
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
Received: from mpg.mcgawpark.baxter.com (159.198.180.50) by ccmailgw.mcgawpark.baxter.com with SMTP (IMA Internet Exchange 2.1 Enterprise) id 0006F10E; Fri, 7 Feb 97 14:10:10 -0600 Received: from sparc5.microscopy.com by mpg.mcgawpark.baxter.com id aa28404; 7 Feb 97 14:10 CST Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id HAA18943 for dist-Microscopy; Fri, 7 Feb 1997 07:51:02 -0600 Received: from THE-SPA.COM (bbs.the-spa.com [204.97.227.2]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id HAA18940 for {microscopy-at-sparc5.microscopy.com} ; Fri, 7 Feb 1997 07:51:01 -0600 with SMTP (IPAD 1.51) id 5043200 ; Fri, 07 Feb 1997 08:59:39 EST X-Sender: ebs-at-ebsciences.com X-Mailer: Windows Eudora Light Version 1.5.2 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii" To: microscopy-at-Sparc5.Microscopy.Com
Dear Terry, In reply to your questions about EPMA: 1. How can it be a "standard" if the composition is "nominal". Get a standard. Use pure element by preference. Also, results from an EPMA will always differ from a bulk technique because they test very different things. Assume they are wrong. Test all the elements in the sample at every point, as there are strong interferences for Cr in Fe. 2. The main problem with carbon analysis in the EPMA is that as you sit on a location trying to get a carbon reading, the microscope is laying down carbon, probably in far greater amounts than are in your standards. I doubt if your detection limit is high enough for the levels you are trying to test. Light element analysis is very sensitive to the matrix, which you do not specify. You wrote: } 1. I have a sample composed primarily of Fe and Cr. I'm } characterizing the compositional Cr gradient as a function of location } on the sample cross-section by EPMA. " Polished surface". My results } for Cr concentration are nearly 20% lower than is expected based on } analyses done in other labs by other techniques of which I have little } or no knowledge of how it was done. I have reports that make } compositional claims but give little or no methodology. } } I'm using a 304 SS standard "nominal 18% Cr" for standardization. On } my sample at a specific location where it is expected to measure 40 } wt % Chromium, I'm getting in the range 30 to 35 Wt % Chromium. } } As I said above this is the bare bones of the circumstance. Any input } regarding Cr/Fe EPMA analysis will be appreciated. } } ON ANOTHER FRONT } } 2. I'm toying with carbon analysis by EPMA. I've got a set of 6 carbon } standards that with the exception of carbon concentration, are } basically 52100 material. Their carbon ranges from 0.018 to 0.610 and } they are very homogeneous. I'm using them to generate a curve from } which I can use to pick off x-ray on peak count levels that relate to } count rates from an unknown. I'm having trouble reconciling count } differences between my curve generating standards and those of a } SRM1225 which contains 0.275 carbon. All the standards have been } verified by Spectrographic analysis. I've run as high as 300 points } on the 1225 material and it consistently gives me lower average counts } than would be expected based on the carbon curve. Regards, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
Dear James, Wood preparation for the SEM is usually by slicing with a razor blade. Make a fairly small block (less than 6 mm. on a face), since all wood outgasses. The face you want to look at should be done last, with a new blade. Some woods are best cut dry, others after soaking or even boiling to soften. My experience is to cut softwoods dry and hardwoods wet. Carbon coat as usual and analyse as usual in the EPMA. The wood is quite beam stable. I have had good luck tracing brominated glues and wood preservatives diffusing into wood. You wrote: } I have been asked about the possibility of measuring elements in tree core } using the electron probe. How best to prepare the wood for 10 - 20 kV, 20 } nA , 30 sec. beam exposure? My only experience with wood usually involves } an axe and a fireplace. Thanks in advance. } } *.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.* } James J. McGee (jmcgee-at-sc.edu) } Dept. of Geological Sciences } University of South Carolina (803) 777-6300 (Office) } Columbia, SC 29208 (803) 777-6610 (Fax) Good luck, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
Dear Jon, I thought only Canadians would be that hard up! The main problem is that the Kevex pre-amp puts out a very different signal than the Link PP is built to receive. Also, different detectors require different bias voltages, so you have to be sure to supply the right one. You can probably come from the Kevex PP out to the Dapple. Do you have the computer and Dapple software? The other solution is IXRF, who are supplying computer systems for working Kevex detectors. You really need a EDX tech type. You wrote: } Over the past few years I have been collecting the components of an EDS } system for our TEM. I think I have all the parts, and now hope to put them } together in the most efficient and cost effective manner (I also want it to } work!). } } Here is what I have: } } Kevex detector to fit our microscope. } } Kevex 7000 chassis with 4505P pulse processor, bias power supply is } somewhere inside, no Kevex software, dead disk drive and rest of system } appears to be dead too. } } NIM bin with another 4505P and PGT bias power supply modules. } } Link model 1134 bias power supply and PP, newer would like to use this if } possible. } } Dapple X-Mate MCA and acquisition controller. } } A Link detector that does not fit our microscope. } } } Questions: } } If I can figure out the plugs and sockets, could I use the Link PP and bias } on the Kevex detector? I would like to do this because the Link box is } newer and a better shape than the Kevex 4505P either in a NIM bin or in the } old Kevex 7000 chassis. } } What is the chance of finding out the pin outs and adjustments needed to } mate the weird combination of plugs and pins I will end up with in a hybrid } system? } } How nuts do you think I am for to try to piece together a system this way } as opposed to just getting all the components from a single source? } } We are kind of Mac oriented in the lab, so I was thinking of a Mac MCA } (maybe 4pi) board to collect and work over spectra once I get some of the } detector/bias/PP part worked out. } } What are some of the other pitfalls I should watch out for in putting } something like this together? We might have as much as $10K to devote to } this project, that would have to go for the new 4pi or other board and the } modifications to the components. } } Any ideas for a better plan? Anybody want the leftovers if it works? Good luck, you'll need it. Regards, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
} A colleague has had difficulty handling Ni grids because of the } high degree of static electricity -- one of the side effects of } the dry laboratory air during Feb. in the Great White North. } He has shunned his woolen sweaters, has been using appropriate } tweezers and when staining has beenn wetting the forceps to } reduce the problem. The critical step is of course when inserting } the grids into the holder for viewing in the TEM. Short of } purchasing an anti-static device, are there any ingenious tips } to overcome the case of the leaping grids? Thanks. } Static? What's that? I've read about it, but we rarely experience it here. HOWEVER, we do frequently have the problem of Ni grids sticking to and otherwise acting funny around forceps. It's magnetism. In fact, I have to run my grids through a demagnetizer before putting them in the TEM or I get horrible astigmatism. Give it a try!
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Becky Holdford wrote: =================================== Can anybody recommend a good hand-held diamond scriber?
I need something pretty robust but with small tip radius. I used to use a scriber made by Fisher Scientific, but they stopped selling them. I mainly scribe silicon wafers with varying amounts of metal and oxide layers. =================================== A nice hand held diamond scribe can be found on our website, given below. It is retractable, "refills" are available, and is inexpensive and in wide use in EM labs.
Disclosure: We believe this is a pretty good choice but then again we are selling them.
Chuck
===================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ =====================================================
This is a forwarded message requested by Profs. Rodney C. Ewing and Lumin Wang at University of New Mexcio.
VACANCY ANNOUNCEMENT
TRANSMISSION ELECTRON MICROSCOPY ANALYTICAL ELECTRON MICROSCOPY -- LABORATORY MANAGER/RESEARCH SCIENTIST
Applications are invited for the position of laboratory manager/research scientist (Research Scientist III) supporting the transmission electron microscopy facilities in the Department of Earth and Planetary Sciences, University of New Mexico. The laboratory includes a JEOL 2010 high resolution TEM and JEOL 2000FX analytical STEM. Both instruments are equipped with EDS capabilities. More information on the lab may be obtained at HTTP//TEM.UNM.EDU.
We hope to fill this position by 1 May, 1997. The position is full time initially for 15 months and may be continued on a permanent basis. Duties will include supervision of all aspects of the electron microscopy laboratory, maintenance of the instruments, assistance to students, faculty, and research scientists at UNM, and outside users, and instruction of a graduate course in principles of electron-microscopy and use of the instruments. Time will be available for independent or collaborative research involving the use of the microscopes and other analytical facilities in the Department.
Candidates must hold a Ph.D. degree in earth science, materials science or a related field, and have a demonstrated research background in these disciplines, extensive skills in the operation and maintenance of transmission electron microscopes, and a record of published research activity. In addition, the candidate should have strong communication skills and an ability to work with and/or instruct individuals with broad research interests and backgrounds.
JOB TITLE: RESEARCH SCIENTIST III DEPARTMENT: EARTH AND PLANETARY SCIENCES REQUISITION NUMBER: 970231*A CLOSING DATE: 5:00 P.M. ON 3/21/97 GRADE 13
Based on Full-Time Salary: $2,858.42 to $3,801.42 mo. Full-time term fifteen month position with possibility of regular status.
IN ORDER TO BE QUALIFIED YOU MUST HAVE:
Ph.D. Degree in Technical, Scientific, or Engineering. Three (3) years experience directly related to the duties and responsibilities specified.
TO APPLY
Applications must be received by the Human Resources Office at 1717 Roma NE, or Health Sciences Ctr., Med Bldg. 2, Rm. 101, North Campus, Albuquerque, NM 87131 no later than 5:00 p.m. on the closing date, February 21, 1997. Resumes must list employment dates by month/year and must be accompanied by a cover letter. Functional resumes will not be accepted. Indicate the requisition number 970231*A and job title Research Scientist III on the application/cover letter. Application forms may be obtained by calling 505-277-6422.
Carolyn J. Emerson wrote: ========================================== A colleague has had difficulty handling Ni grids because of the high degree of static electricity -- one of the side effects of the dry laboratory air during Feb. in the Great White North. He has shunned his woolen sweaters, has been using appropriate tweezers and when staining has beenn wetting the forceps to reduce the problem. The critical step is of course when inserting the grids into the holder for viewing in the TEM. Short of purchasing an anti-static device, are there any ingenious tips to overcome the case of the leaping grids? ========================================== I would like to clarify some misunderstandings about tweezers and their "nonmagnetic" nature. First, the so-called "nonmagnetic" or "anti-magnetic" stainless steel tweezers are not 100% anti-magnetic. It is my understanding that the very best antimagnetic stainless steel (the kind that is used for Dumont, SPI and other major manufacturer's of Swiss tweezers) is 92-95% antimagnetic maximum. And that alone is enough residual magnetism to cause nickel grids to stick to the so-called nonmagnetic tweezer tips. While antistatic devices may or may not reduce the effect, the point is most of the problem is caused by residual magnetism in the tweezer tips.
The SPI "Miracle Tip" and "Gold Plated Miracle" tip tweezers are made of a super allow (it is not stainless steel at all) that really is 100% antimagnetic. Nickel grids will not stick to the tips of these tweezers. The gold plated version of the product keeps the low pH of the typical reaction from reacting with the metal (e.g. electrochemistry), thereby stunting the strength of the reaction. Additional information about these tweezers can be found in the SPI On-Line catalog given below.
Disclosure: SPI has offered for some years tweezers with tips that are 100% antimagnetic and are ideal for working with nickel grids, especially for immunogold work.
Chuck ====================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ =====================================================
Jeff Fortner wrote: } } I've seen a couple of responses attributing this effect to reflected } electrons. Although I've never observed these "reflection" images (we } religiously coat our glass samples, although I understand why one may not want } to coat an art relic), I think I have a better explanation. The surface of the } non-conducting sample is acting as part of a capacitor... the other part being } the inside of the chamber. What you are imaging is a surface charge set up by } this capacitance, and not reflected electrons. Electrons deflected completely } away from the sample are unlikely to image much of anything (try running the } EDS simultaneously with this effect...if the electrons are reflected there } should be a huge bremstraalung peak, and nothing else. } ........................... Gotta disagree with you on this one Jeff,
The idea of a surface image charge is interesting, but doesn't correspond to the characteristics of the phenomenon. Let me state several distinctive characteristics:
1. You can focus these images just like an ordinary specimen image. 2. The topography of the reflecting "mirror" specimen disappears. 3. One can image and differentiate objects which are at a uniform ground potential. 4. One can image objects which are quite some distance away. 5. The effect requires one to first "charge up" the surface at a higher beam voltage.
Let me illustrate by my first encounter with this effect. I was imaging on a swatch of nylon fabric at 1 keV when a local thunderstorm knocked the power out. When power came back on, the SEM came back online at 30 keV and after tuning up my filament settings, I attempted to return to imaging my nylon material at 1 keV (not very bright in retrospect, but hey, it was 3 a.m. and I wasn't too coherent). Instead of seeing the fibrous structures I had seen earlier (at the same relatively low mag) I was seeing unexpected unfocused contours -- some rounded, some linear -- vaguely familiar, but I couldn't put my finger on it. Fiddling with the controls, I noticed that by focusing to longer working distances, the shapes became sharper -- but still in no way recognizable as the material under the beam (I had meanwhile peeked through the glass viewport and verified that the nylon sample really WAS what was under the beam). As the image came into sharper focus, I suddenly realized that what I was seeing was a "fisheye" image of the entire inside of the SEM chamber -- polepiece, detectors, stage, and other internal mechanisms. For a moment I felt like I had entered some sort of "Twilight Zone"!
After I figured out what was going on I became enamored with the effect for a time and perfected my technique -- graduating to a saphire bead with which I could make truly detailed and relatively distortion-free images of the chamber interior. I could easily zoom in on and image parts of the chamber which were 12-15 inches from the specimen (this was a very large chamber).
The point is that although the nylon material had an obviously irregular topography, it produced a quite uniform "mirroring" field. This is not surprising, since the electric potential of an insulator charged up by 30 keV electrons will deflect a 1 keV electron at some distance from the surface. At this distance the contributions from the various surface charge sites integrate into a quite uniform equipotential surface which acts as a nice smooth mirror for the incident electrons. The incident electrons "bounce" off the equipotential surface according to the normal laws of optical reflection and the electron trajectories behave exactly as if one introduced a mirror in the path such that the scanning beam now scans the objects visible in the mirror -- the interior of the specimen chamber. It takes only a very weak field to attract the produced secondaries to the detector so a usable image is produced (though typically weaker, given the longer collection distances.)
If a capacitive "image charge" were responsible, one would need to have a very regular capacitor surface to retain an intelligible image -- clearly not the case with the nylon swatch. Secondly, creation of such an imaging charge would require that the features being imaged would need to be producing strong variations of local field at the capacitor surface -- not the case when I could image features of the chamber which were all at ground potential and at considerable distance. Finally, an image charge would not lend itself to focusing.
I can see the merits of your hypothesis, especially when the original question involved seeing the secondary detector (which is at an elevated potential) on a glass surface (presumably smooth). I can imagine a weak image charge being produced on the glass via the proximity of the SED field. But this would be evidenced by a rather subtle and smooth modulation of the normal image contrast (remember that the SED collection field at the specimen is necessarily weak and uniform, else the incident beam would also be badly deflected). In the case of the effect I have been talking about, the topography of the specimen is REPLACED with a highly detailed reflected image -- exactly as if a mirror were inserted above the specimen.
I don't recall ever attempting to look at the x-ray spectrum produced. I wouldn't expect to see anything much since the electrons are striking objects which are out of the line of sight of the EDS detector. A pure brehmstrahlung spectrum should be produced, as you suggest, and this is an interesting idea.
A final note -- in my earlier posting I commented that I knew of no good use for this effect. In fact, I did once use it to locate a breakdown across an interior insulator. It is also a good way of noting which interior features of the chamber are most strongly producing secondary electron "background" as would normally occur from electron backscattering onto the chamber walls. But its best use IMHO is still its considerable potential for amusement!
Ni grids can be a pain, especially the first time they are used by those who have only used Cu grids, because they are magnetic. Furthermore, I have found that many so-called non-magnetic forceps don't seem to be adequately non-magnetic. In over 10 years experience the ones I recommend are Dumont INOX, in the common tip type and self-closing - N5. I have no financial connection to Dumont (I wish I did!). Bruce Cutler, Microscopy Lab, University of Kansas, Lawrence
Greetings, For a low-tech solution to excess static, we keep a box of "Bounce free" squares in the lab. These squares are made to put in a load of clothing in the dryer and prevent wrinkles. The "free" in the name means that they are free of fragrance. You can put one of these squares on the surface and work over it (i.e., put a dish on the square). You can wipe off areas or objects. Keeps the static charges down. I wipe my computer monitor with one and dust stays away for a long while. We buy these things in our local supermarket. Obviously, this won't do anything for residual magnitism. I have no stake in the Bounce company. Hope this helps, Tobias
I wouldn't have thought the Ni grids are being affected by static as much as magnetic fields due to magnetised forceps tips. To alleviate this try demagnetising by scrambling the domains in a high field strength such as in an old mains transformer with a cut out channeled in the metal pole piece. This works for us and is a cheap solution but I dare say someone will sell you a "degausser" which will work equally well and might look more acceptable to the safety officer ! Ask your physics/electronics people for a supplier.
Regards
Laurence Tetley
At 16:01 07/02/97 -0330, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
A few days ago I put in the following request:I am in need of acquiring the conical part of the final lens of a JSM 35(version C) scanning electron microscope. The cone in question is the part of the lens that hangs down in the chamber. We intend to develop a modification. Please email me with the information so that further negotiations can be carried out. I should welcome also any information on the composition (and commercial specification) of the metal from which the cone is made of. Of course, info on where this metal can be obtained will be invaluable.
Thank you in anticipation of your cooperation and help.
Although I have had two replies, my quest continues. If you have any suggestions/comments, please do not hesitate to get in touch.
Many Thanks.
Jitu Shah
Dr.Jitu Shah H.H. Wills Physics Laboratory, University of Bristol, Royal Fort, Tyndall Avenue, Bristol BS8 1TL. UK email: jss-at-siva.bristol.ac.uk Tel: 44 117 9288719 Fax: 44 117 9255624
Has anyone had experience staining tissues (breast, etc.) for the presence of silicone gel at either EM or Light levels? As far as I know, the silicone does not accept stain but thought I would learn if anyone has had any experience with such specimens.
Could someone give me information concerning where to purchase inexpensive silicon wafers of any diameter and about 0.5 mm thickness. Silicon quality is not important. I need these wafers to sandwich cross-section TEM samples. Thank you in advance.
} Is someone familiar with a paper or book which } can be used to determine the distance necessary between adjacent } serial sections, given the size of structures that I am attempting to } join up between sections, for a given statistical significance? } } Any information would be greatly appreciated.
You might try "Stereological methods" by Ewald R. Weibel, Academic Press, 1980. It is better known to people who do morphometry as "Weibel's Bible". The first volume is practical stuff with examples (mostly from biologic science) the second volume is theorectical stuff. Unfortunately, someone has borrowed my copy so I don't know for sure that what you want is in there, but that's where I'd start.
Leon
-- Leon A. Metlay, M.D.,Associate Professor of Pathology and Laboratory Medicine University of Rochester Medical Center Phone: (716) 275-5691 P.O. Box 626 Fax: (716) 273-1027 Rochester, NY 14642 lmetlay-at-acu.pathology.rochester.edu http://www.urmc.rochester.edu/smd/pathres/URPLM.html "Most ass drivers are evil, most camel drivers are decent, most sailors are saintly, the best among physicians is going to Gehenna, and the best of butchers is a partner of Amalek" -R. Judah, in Mish. Kidd. 4:14
Hi, I hope someone there can give a hand for education of microcopy to our young generation.
*********************************************** * Ming H. Chen, PhD * * Medicine/Dentistry Electron Microscopy Unit * * University Of Alberta. * * Edmonton, Alberta, Canada * * * * Visit My Page At: * * http://www.ualberta.ca/~mingchen * ***********************************************
---------- Forwarded message ----------
We have a few thousand scrap 4" test wafers that we sell for $1 each in quantities of 50 for just this kind of use.
best regards mark
Mark W. Lund, PhD Director } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"367 billion dollars of coal clearly has 367,000 times the value of a million dollar view."
Some time back I posted a request for help. The response I received was GREAT!
Now, maybe, I can be of some help to you. I have created a web page with about 90 (so far) LINKS to companies and other interesting sites. I hope this will be of value to those who are looking for information.
Please, let me know if I have made any errors or omissions. I will be happy to make changes or add new sites.
Oh, yes, the MSA gets top billing!
Again, thank you for being there and for all the help and on-going information.
Best regards,
Bob
E-mail: bobcat54-at-aol.com Home Page: http://members.aol.com/BobCat54/index.html Springfield, MA, USA
Some time ago I jumped on the digitised image bandwagon. In cases where a digital image is not acquired at the start, I scan the negative using an Agfa Arcus II scanner (recommended by folks on this listserver).
My problem is that in some instances, I get a pattern on my images, reminiscent of thickness fringes, which I am interpreting as some sort of Moire effect. I've purchased a few of these scanners now, for different areas, and the problem occurs to varying degrees on all of them. On the unit which is most prone to this, the transparency module is visibly crooked, which may be the cause. I do get the problem on the others, though, and the lid appears quite straight on them.
I've yet to find the appropriate Agfa contact who can help, so if one is out there, or if any one else can shed some light on this for me, I'd appreciate it. I don't want to go back to the darkroom!
**************************************** Don Steele Steele-at-KRDC.INT.Alcan.Ca Alcan International Kingston Research and Development Centre (613) 541 - 2145 ****************************************
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The best way to estimate section thickness is to look at folds in the section. Small enough folds should be x2 the section thinckness.
Alternatively, it is possible to re-embed the grids with sections on to obtain cross sections from which you can directly measure the section thickness. In this case is it interesting to see how variable the section thicknesses are.
An exotic way to estimate section thickness is to trim the block at a pre-determined angle and measure the increasing dimensions of the block as the sections are removed. I lost my high school geometry book so I can't help you more on this one.
Paul Webster, Ph.D. Center for Cell Imaging http://info.med.yale.edu/cellimg
Nestor Zaluzec Your Friendly Neighborhood SysOp --------------
} Fellow microscopists, } } I am trying to locate a copy of the NIH Image Processing Software. on the } net. Can some one please forward the respective address. } } } Thank you } Mitch } Evex Analytical } } } } } Evex is the world's leading independent provider of service and support for } Evex, Link, Kevex, Noran, P.G.T. and Tracor brand EDS systems and related } peripherals. Our Service Engineers are "factory trained" and are located } nationwide.
To alleviate this try } demagnetising by scrambling the domains in a high field strength such as in } an old mains transformer with a cut out channeled in the metal pole piece. } This works for us and is a cheap solution but I dare say someone will sell } you a "degausser" which will work equally well and might look more } acceptable to the safety officer ! Ask your physics/electronics people for } a supplier. } } Regards } } Laurence Tetley } A degausser that you can buy and perhaps use for something else is a soldering gun that has the two leads coming out to form the tip. I think that Sears sells one like that that has a light on it. Put your tweezers slowly in and slowly out and it will degauss them. - -Scott Walck
I have a what would appear to be a simple question. In the case of dislocations in a two beam condition, one gets contrast at dislocations, and of course the reverse contrast in dark field. I understand the imaging conditions regarding the strain field and g as to when contrast should occur. I am curious if there is a simple explanation as to why in the vicinity of dislocations which are after all spatially localized events in real space and hence delocalized in Fourier space as to why dislocations diffract more into the diffraction spot (e.g. why are dislocations bright in bright field (as compared to the background). Sorry for asking what must be obvious to all, but I am self taught from TEM textbooks and if possible would like a simple picture in addition to the math. Is the answer essentially dynamical in that the dislocation causes scattering off other band in the dispersion surface than the two beam case, and if so why then is the two beam dark field bright?
Thanks for an answer to a silly question, Paul Fons
Dr. Paul Fons Senior Scientist Electrotechnical Laboratory Tsukuba, Japan 305
fax: 81-209-58-5615 tel: 81-298-58-5636
email: fons-at-etl.go.jp Eudora Enclosures O.K.
PGP Public Key -----BEGIN PGP PUBLIC KEY BLOCK----- Version: 2.6.3i
I have a what would appear to be a simple question. In the case of dislocations in a two beam condition, one gets contrast at dislocations, and of course the reverse contrast in dark field. I understand the imaging conditions regarding the strain field and g as to when contrast should occur. I am curious if there is a simple explanation as to why in the vicinity of dislocations which are after all spatially localized events in real space and hence delocalized in Fourier space as to why dislocations diffract more into the diffraction spot (e.g. why are dislocations bright in bright field (as compared to the background). Sorry for asking what must be obvious to all, but I am self taught from TEM textbooks and if possible would like a simple picture in addition to the math. Is the answer essentially dynamical in that the dislocation causes scattering off other band in the dispersion surface than the two beam case, and if so why then is the two beam dark field bright?
Thanks for an answer to a silly question, Paul Fons
Dr. Paul Fons Senior Scientist Electrotechnical Laboratory Tsukuba, Japan 305
fax: 81-209-58-5615 tel: 81-298-58-5636
email: fons-at-etl.go.jp Eudora Enclosures O.K.
PGP Public Key -----BEGIN PGP PUBLIC KEY BLOCK----- Version: 2.6.3i
I have a what would appear to be a simple question. In the case of dislocations in a two beam condition, one gets contrast at dislocations, and of course the reverse contrast in dark field. I understand the imaging conditions regarding the strain field and g as to when contrast should occur. I am curious if there is a simple explanation as to why in the vicinity of dislocations which are after all spatially localized events in real space and hence delocalized in Fourier space as to why dislocations diffract more into the diffraction spot (e.g. why are dislocations bright in bright field (as compared to the background). Sorry for asking what must be obvious to all, but I am self taught from TEM textbooks and if possible would like a simple picture in addition to the math. Is the answer essentially dynamical in that the dislocation causes scattering off other band in the dispersion surface than the two beam case, and if so why then is the two beam dark field bright?
Thanks for an answer to a silly question, Paul Fons
Dr. Paul Fons Senior Scientist Electrotechnical Laboratory Tsukuba, Japan 305
fax: 81-209-58-5615 tel: 81-298-58-5636
email: fons-at-etl.go.jp Eudora Enclosures O.K.
PGP Public Key -----BEGIN PGP PUBLIC KEY BLOCK----- Version: 2.6.3i
1997 ADVANCED SPECIMEN PREPARATION WORKSHOPS (for LM, SEM/TEM, and SPM)
The following 8 independent workshops offer an intensive hands-on training program for the application of the most advanced specimen preparation techniques currently available for microscopy of complex material systems. These workshops are intended for R&D personnel involved in microscopy of advanced materials and/or related specimen preparation. Enrollment is limited to 4 students in each workshop and early registration is strongly recommended to ensure admission.
***Site-specific Cross-sectioning and Microthinning Precision Cleaving (May 6 or Nov. 4 in Santa Clara, CA) FIB-milling for SEM Cross-sectioning (May 7 or Nov.5 in Sunnyvale, CA) FIB-milling for TEM Cross-sectioning (May 7-9 or Nov.5-7 in Sunnycale, CA) Precision Lapping for SEM Cross-sectioning (May 14 or Nov. 12 in Phoenix, AZ) TEM Wedge-polishing (May 14-16 or Nov. 12-14 in Phoenix, AZ)
***Materials Ultramicrotomy General Surface Preparation for LM, SEM, or AFM (May 19-20 or Nov. 17-18 in Phoenix, AZ) Thin Section Preparation for TEM (May 19-21 or Nov. 17-19 in Phoenix, AZ) Advanced Ultramicrotomy (May 22-23 or Nov 20-21 in Phoenix, AZ)
A partial list of the past participants in these workshops include: IBM, Motorola, SEMATECH, Texas Instruments, Medtronic, Hewlett-Packard, Lawrence Berkeley Lab, Oak Ridge National Lab, National Renewable Energy Lab, Bell Northern Research (Canada), Honeywell, Martin Marietta, B.F. Goodrich, 3M, MIT Lincoln Lab, United Technologies, Hydro Quebec (Canada), Cabot, Lawrence Livermore National Lab, US Army Research Lab, Kimberly Clark, etc.
For further information and on-line registration, please see our home page hosted on Microscopy Online at http://www.microscopy-online.com/Vendors/AMCGroup/. You may also request a copy of the workshops brochure by providing us with your complete mailing address.
Rene E. Nicholas AMC Group (a Division of Promotech Associates, Inc.) amcgroup2-at-aol.com
} Is someone familiar with a paper or book which } can be used to determine the distance necessary between adjacent } serial sections, given the size of structures that I am attempting to } join up between sections, for a given statistical significance? } } Any information would be greatly appreciated.
You may try Aherne, W.A. & Dunnill, M.S., 1982. Morphometry, Edward Arnold, London.
A number of papers on "stereology" have been published by Gundersen, H.J.G. and co-workers. Take a look in Acta Pathol. Microbiol. Immun. Scand. (APMIS) vol 96.
You are indeed seeing Moire patterns from the scanned images. This is due to the dot pattern in the image and it is a real problem with glossy images taken from journals or textbooks. There are several ways around the problem. One way is to play with the dpi setting of your scanner and find a resolution that minimizes the pattern. Another is to scan the image in at a high resolution and do a 1-2 pixel gaussian blur of the image or to adjust the dpi setting down from a high resolution (600 dpi) to a lower resolution (100-200 dpi). These latter solutions may be performed in an image processing program like Photoshop. You can also use fast-fourier transforms to remove the patterns, but I am not as pleased with the results.
Regards,
John J. Turek, Ph.D. Purdue University Dept. of Basic Medical Sciences Director, Core Laboratory for Image Analysis and Multidimensional Applications (CRISTAL) phone: 317-494-5854 fax: 317-494-0781 email: jjt-at-vet.purdue.edu web: http://vet.purdue.edu/cristal
Postdoctoral Positions in Electron Microscopy Brookhaven National Laboratory
A postdoctoral opening is available in the Materials Science Division, Department of Applied Science at Brookhaven National Laboratory. The appointment is for one year initially with the possibility of renewal for longer terms, and involves the use of BNL's new JEOL 300kV FEG transmission electron microscope in studying high-temperature superconductors, hard magnets, and other materials of interest. This state-of-the-art instrument has a point-to-point resolution of 0.16nm, energy resolution of 0.65eV, equipped with multiscan CCD cameras, a Gatan Imaging Filter, an electron energy-loss spectrometer, an energy-dispersive x-ray spectrometer, and a holography unit. The attached scanning system can provide a {0.2nm probe with a x-ray chemical-mapping resolution of 1nm, and has an annular-dark-field detector with Z-contrast imaging capability. Heating and liquid helium stages will also be available.
The successful candidate will be a recent Ph.D graduate in physics or materials science with a strong background in structural analysis, as well as in electron microscopy. Research experience in crystal structure, structural defects, and interfaces using electron-microscopy imaging, diffraction, including diffuse scattering, spectroscopy, GIF, holography, and computer simulation is desired. Qualified candidates should send their resume and names and addresses of three referees to :
Dr. Yimei Zhu Building 480, Materials Science Division, Brookhaven National Laboratory Upton, Long Island, NY 11973-5000 U.S.A.
phone: (516) 344-3057 fax: (516) 344-4071
BNL is a multipurpose national laboratory managed by Associated University Inc. for the U.S. Department of Energy. BNL is an equal opportunity employer committed to build and maintaining a diverse work force.
******************************** Dr. Yimei Zhu Materials Science Division Brookhaven National Laboratory Upton, Long Island, NY 11973 USA Tel. (516)344-3057 Fax. (516)344-4071 ********************************
If the Windows 95 version (not beta) is not out yet, it should be shortly. It is being developed by Scion Corp. They have more info at their web site, the address of which I don't have handy right now. The NIH image www site also has additional info. ------------------------------------------------ Opinions or statements expressed herein, rational or otherwise, do not necessarily reflect those of my employer.
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 Phone: (508) 750-1717 E-mail: crossman-at-osi.sylvania.com
Our web sites: www.sylvania.com www.siemens.com --
At 11:49 AM 2/8/97 -0500, Fred Schamber wrote: } I don't recall ever attempting to look at the x-ray spectrum produced. } I wouldn't expect to see anything much since the electrons are striking } objects which are out of the line of sight of the EDS detector. A pure } brehmstrahlung spectrum should be produced, as you suggest, and this is } an interesting idea. } Yes, one would hope and expect to see few x-rays then. However, some are detected and they are not all bremsstrahlung:
Unless fast electrons reach the atoms in the sample they cannot generate bremsstrahlung in the sample. The electrons reflected by a strong enough electrostatic field produced by an electrically isolated, electrically charged specimen can generate characteristic x-rays as well as bremsstrahlung from all specimen chamber materials which are visible in the mirror image of the chamber.
If an abnormal background shape is observed in x-ray spectra recorded in the "mirror" condition, a significant source is the reflected electrons themselves -- either entering the x-ray collimator and generating detectable x-rays there or even penetrating the detector window and reaching the detector crystal itself (especially if there is a thin window or if there is no magnetic "electron trap" in the collimator). BTW, this same effect can be observed clearly in TEM or STEM if the collimator's internal geometry is bad and the EM is operated with a very low or zero magnetic lens field at the specimen, as is commonly found in low mag mode.
Best wishes, Brian
Brian W Robertson Office 402 472 8308 Associate Professor Lab 402 472 8762 Department of Mechanical Engineering and FAX 402 472 1465 Center for Materials Research and Analysis, University of Nebraska-Lincoln 255 Walter Scott Engineering Center Lincoln NE 68588-0656 USA
There is a 32 bit Windows 95 version of NIH Image at http://rsb.info.nih.gov/nih-image/download.html The Web page says that it is for Windows NT also, but according to Scion Corp., the NT version is scheduled for release sometime in the Spring. Stanley L. Flegler Center for Electron Optics Michigan State University flegler-at-pilot.msu.edu
Many thanks to all who replied concerning the difficulty in handling Ni grids. Several folk correctly pointed out that in addition to difficulties in handling grids in general if there is static involved (and there is in our lab with the heat cranked up in winter), Ni grids create an extra problem because of their ferromagnetic nature and attraction to non-magnetic forceps tips.
Advice ranged from dealing with the possibility of static by working over an area covered with Bounce Free anti-static laundry sheets, to making minor modifications of specimen loading ports on the TEM. Several people commented that even those forceps described as non-magnetic were not totally so, and could attract Ni grids. There was advice to dip the tips in glacial acetic acid and wipe dry, or rinse in ethanol. Others talked of degaussers or demagnetizing devices available from one's local physics dept or electronics shop to demagnetise forceps and/or grids. A vendor did direct us to truly non-magnetic gold-tipped forceps which are available commercially.
And several microscopists pointed out the additional problem of astigmatism in the microscope while viewing Ni grids and suggested we save ourselves all the grief and use gold grids when doing immuno work.
My colleague is going to purchase some gold grids, we've got a box of Bounce sheets on the lab bench, and we'll also investigate demagnetisers and the truly non-magnetic forceps.
Thanks to all who responded on the listserver and privately.
Carolyn J. Emerson email: cemerson-at-plato.ucs.mun.ca
Biology Department Memorial University St. John's, NF A1B 3X9 Tel: (709) 737-7515 Fax: (709) 737-3018
PPG Glass Technology Center located 14 miles Northeast of Pittsburgh has an immediate opening in its Electron Microscopy Laboratory.
The position interacts with R&D, manufacturing and technical support personnel in designing analytical approaches and providing meaningful solutions to projects relating to glass, glass coatings, paints and many other PPG manufactured or formulated products.
The candidate will be required to operate a JEOL STEM and a Digital scanning probe microscope, supervise a group of 4-5, and manage this well-equipped microscopy laboratory.
The position requires an advanced degree in materials science, physics or chemistry, or the equivalent in relevant experience. The candidate must have 3 or more years of industrial experience. In-depth knowledge of STEM, SEM, and experience in data/image handling by computers is essential. Experience in surface analysis, IR spectroscopy and other analytical techniques is desirable.
The candidate must have technical and organizational skills, must be able to work independently and must have the interpersonal skills to work effectively with others. Official authorization to work permanently in the United States is required.
Please forward resume and salary information to:
PPG Industries P.O. Box 11472 Pittsburgh, PA 15238
PPG Glass Technology Center located 14 miles Northeast of Pittsburgh has an immediate opening in its Electron Microscopy Laboratory.
The position interacts with R&D, manufacturing and technical support personnel in designing analytical approaches and providing meaningful solutions to projects relating to glass, glass coatings, paints and many other PPG manufactured or formulated products.
The candidate will be required to operate a JEOL STEM and a Digital scanning probe microscope, supervise a group of 4-5, and manage this well-equipped microscopy laboratory.
The position requires an advanced degree in materials science, physics or chemistry, or the equivalent in relevant experience. The candidate must have 3 or more years of industrial experience. In-depth knowledge of STEM, SEM, and experience in data/image handling by computers is essential. Experience in surface analysis, IR spectroscopy and other analytical techniques is desirable.
The candidate must have technical and organizational skills, must be able to work independently and must have the interpersonal skills to work effectively with others. Official authorization to work permanently in the United States is required.
Please forward resume and salary information to:
PPG Industries P.O. Box 11472 Pittsburgh, PA 15238
PPG Glass Technology Center located 14 miles Northeast of Pittsburgh has an immediate opening in its Electron Microscopy Laboratory.
The position interacts with R&D, manufacturing and technical support personnel in designing analytical approaches and providing meaningful solutions to projects relating to glass, glass coatings, paints and many other PPG manufactured or formulated products.
The candidate will be required to operate a JEOL STEM and a Digital scanning probe microscope, supervise a group of 4-5, and manage this well-equipped microscopy laboratory.
The position requires an advanced degree in materials science, physics or chemistry, or the equivalent in relevant experience. The candidate must have 3 or more years of industrial experience. In-depth knowledge of STEM, SEM, and experience in data/image handling by computers is essential. Experience in surface analysis, IR spectroscopy and other analytical techniques is desirable.
The candidate must have technical and organizational skills, must be able to work independently and must have the interpersonal skills to work effectively with others. Official authorization to work permanently in the United States is required.
Please forward resume and salary information to:
PPG Industries P.O. Box 11472 Pittsburgh, PA 15238
PPG Glass Technology Center located 14 miles Northeast of Pittsburgh has an immediate opening in its Electron Microscopy Laboratory.
The position interacts with R&D, manufacturing and technical support personnel in designing analytical approaches and providing meaningful solutions to projects relating to glass, glass coatings, paints and many other PPG manufactured or formulated products.
The candidate will be required to operate a JEOL STEM and a Digital scanning probe microscope, supervise a group of 4-5, and manage this well-equipped microscopy laboratory.
The position requires an advanced degree in materials science, physics or chemistry, or the equivalent in relevant experience. The candidate must have 3 or more years of industrial experience. In-depth knowledge of STEM, SEM, and experience in data/image handling by computers is essential. Experience in surface analysis, IR spectroscopy and other analytical techniques is desirable.
The candidate must have technical and organizational skills, must be able to work independently and must have the interpersonal skills to work effectively with others. Official authorization to work permanently in the United States is required.
Please forward resume and salary information to:
PPG Industries P.O. Box 11472 Pittsburgh, PA 15238
I was wondering if someone could recommend one or two commercial labs where we could outsource some of our TEM polymer work. Our samples would require embedding, cryo or RT sectioning, staining (usually phosphotungstic acid ) and a few TEM micrographs.
EM Posting-PPG Please forward resume and salary information to: PPG Industries P.O. Box 11472 Pittsburgh, PA 15238 Att: Supervisor, Personnel AN EQUAL OPPORTUNITY EMPLOYER M/F/D/V
Does anyone out there have information that compares/contrasts the techniques of laser profilometry, contact profilometry, and SEM imaging over wide areas (i.e. 2x2 mm)? The goal is to see if contact or laser profilometry (quant. data) can be substituted for SEM imaging (guesswork and hand-waving) in topographic analysis of layers of 4-10 micrometer generally spherical particles.
The person wants quick and easy turnaround with as little human interpretation as possible.
The things we do for money! ------------------------------------------------ Opinions or statements expressed herein, rational or otherwise, do not necessarily reflect those of my employer.
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 Phone: (508) 750-1717 E-mail: crossman-at-osi.sylvania.com
Our web sites: www.sylvania.com www.siemens.com --
I would be grateful if the email addresses of major microscope manufacturers,optics-eyepiece,objective,phase contrast eqpt,manufacturers dealers could be mailed to me. Regards,
} I went after NIH Image, and found the 0README.txt claims there is } no DOS version, it's only for Mac.
There is an excellent free program for DOS/Win32/Win95 platforms that stands up very well to NIH Image: ImageTool by Don Wilcox, Brent Dove, Doss McDavid and David Greer at the University of Texas Health Science Center in San Antonio:
Find version 1.27 at http://ddsdx.uthscsa.edu/dig/itdesc.html
ChR
_________________________________________________________________________ Christophe Roos Dr.Sc., doc. | Institute of Biotechnology | & Dept. of Biosciences Phone: +358 9 7085 9367 | Division of Genetics Fax: +358 9 7085 9366 | P.O.Box 56, Viksbagen 9 E-mail: Christophe.Roos-at-Helsinki.FI | FIN-00014 Univ. of Helsinki X-400: /G=Christophe/S=Roos/O=Helsinki/ADMD=fumail/C=Fi Finland {A HREF="http://www.helsinki.fi/~roos/index.html"} WWW Home Page: Roos {/A} -------------------------------------------------------------------------
I picked up your e-mail message about getting some microscopy done on polymers.We could do this sort of thing in the multi-Imaging Centre here in Cambridge UK. We have croy-SEM and ED X-ray microanalysis, Cryo-TEM and Cryoultramicrotomes. Collectively we have a 120 years of experience. Our rates are very competative and we can do a fast turn around.
Contact me on e-mail 'Phone +44-1223-333946 or Fax +44-1223-333953.
Patrick Echlin Director, Multi-Imaging Centre. University of Cambridge Cambridge CB2 3EA United Kingdom
PS Happy Lincoln's Birthday
On Tue, 11 Feb 1997, Marti, Jordi wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Hello ! } } I was wondering if someone could recommend one or two commercial labs } where we could outsource some of our TEM polymer work. Our samples } would require embedding, cryo or RT sectioning, staining (usually } phosphotungstic acid ) and a few TEM micrographs. } } Thanks } } Jordi Marti }
To image the inside of my SEM, I first stick a piece of PTFE or a round glass coverslip onto a stub and then glue a small ball bearing (3mm) onto the centre. This gives a much better view of the chamber. You can try using larger ball bearings, also to give a weird effect try sticking two small ball bearings together.
Also try switching on your back scatter detector once you have 'charged' up the stub, you can visualise the sectors - (if + the sector is white and if - the sector is black)
Kevin Mackenzie Tillydrone E.M. Unit University of Aberdeen Tillydrone Avenue Aberdeen AB9 2NT SCOTLAND Tel 01224-272847 Fax 01224-272396
} Hello ! } } I was wondering if someone could recommend one or two commercial labs } where we could outsource some of our TEM polymer work. Our samples } would require embedding, cryo or RT sectioning, staining (usually } phosphotungstic acid ) and a few TEM micrographs. } } Thanks } } Jordi Marti
If you check out the November issue of MICROSCOPY & ANALYSIS, you'll find a listing of some 40 labs in the US offering various microscopy services. If you can't find a copy, contact me and I'll e-mail you the list.
Regards,
--------------------------------------------------------------- Dr L. P. Stoter Technical Editor, MICROSCOPY & ANALYSIS Technesis 17, Rocks Park Road email: LPS-at-teknesis.demon.co.uk Uckfield, E. Sussex Phone: +44 (0)1825 766911 TN22 2AT Fax: +44 (0)1825 766911 United Kingdom ---------------------------------------------------------------
To image the inside of my SEM, I first stick a piece of PTFE or a round glass coverslip onto a stub and then glue a small ball bearing (3mm) onto the centre. This gives a much better view of the chamber. You can try using larger ball bearings, also to give a weird effect try sticking two small ball bearings together.
Also try switching on your back scatter detector once you have 'charged' up the stub, you can visualise the sectors - ( the + sector shows white and the - sector is black)
Kevin Mackenzie Tillydrone E.M. Unit University of Aberdeen Tillydrone Avenue Aberdeen AB9 2NT SCOTLAND Tel 01224-272847 Fax 01224-272396
I wonder if somebody could give me any hints on special embedding and/or staining methods for halite crystals (sedimented from aerosols). This would be helpful for a research project which intends to differentiate halite crystals from other aerosol particles by image analysis.
Thanks in advance Hiltrud Mueller-Sigmund
Hiltrud Mueller-Sigmund (hiltrud-at-ruf.uni-freiburg.de) Institut f. Mineralogie, Petrologie und Geochemie Albertstr. 23b - D 79104 Freiburg i. Br. (Germany) Tel.: (+49)-761-203-6388 / Fax: (+49)-761-203-6407
PPG Glass Technology Center located 14 miles northeast of Pittsburgh has an immediate opening in its Electron Microscopy Laboratory.
The position interacts with R&D, manufacturing and technical support personnel in designing analytical approaches and providing meaningful solutions to projects relating to glass, glass coatings, paints and many other PPG manufactured or formulated products.
The candidate will be required to operate a JEOL STEM and a Digital Scanning
Probe Microscope, supervise a group of 4-5, and manage this well-equipped microscopy laboratory.
The position requires an advanced degree in materials science, physics or chemistry, or the equivalent in relevant experience. The candidate must have 3 or more years of industrial experience. In-depth knowledge of STEM, SEM, and experience in data/image handling by computers is essential. Experience in surface analysis, IR spectroscopy and other analytical techniques is desirable.
The candidate must have technical and organizational skills, must be able to work independently and must have the interpersonal skills to work effectively with others. Official authorization to work permanently in the United States is required.
Please forward resume and salary information to:
PPG Industries, Inc. Glass Technology Center P.O. Box 11472 Pittsburgh, PA 15238
I hope this information is of some help. It's not a complete list, but the information I have is as follows. If there is no e-mail address, just contact the home page. I'm sure all of the manufacturers will have a link for messages.
Leica: No address just for e-mail, but their home page can be found at the URL: http://www.leica.com in the USA. Probably have links to other countries from here.
Zeiss: E-mail to micro-at-zeiss.com Home page is http://www.zeiss.com
Nikon: No e-mail address. Home page is http://www.nikonusa.com If you contact Nikon USA, you will probably find a link to addresses for other countries.
Also, please check out the WWW Directory of Microscopy and Microanalysis Products and Services at: http://www.mwrn.com/product/ You can link directly to the manufacturers of microscopes, cameras, optical components, services, accessories, etc. from this page.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
To: roos-at-operoni.helsinki.fi Microscopy ListSer {Microscopy-at-MSA.Microscopy.Com} cc: VANHART --IBMUSM00 Van Hart, D.C. *** Reply to note of 02/12/97 00:58
I FTP'd these files, and found they need Win95 or WinNT to run. Win-OS/2 can't do the install. The site was;
Our lab has an opportunity to pick up some work doing clinical specimens for $$. Currently we are strictly a research facility, but in today's world some cash can really go a long way to justify our existence. Can anyone advise us as to the "Chicago-style pot holes" that may be looming unforseen on the horizon? Do we need clinical accreditation? How much should we charge for clinical work? Any thoughts about turn around time? QC? Are there things we should get straight and in writing before we begin? Thanks for the input.
p.s. Many Thanks for the Alizarin Red stuff....it's been tried and is working great!!
Linda Fox Loyola University Medical School Maywood Illinois lfox1-at-wpo.it.luc.edu
Dear Friends, I have a thin (1um) al2o3/al film on an aluminum (1mm) substrate. I am trying to mount for the cross section. I need a method to preserve the edge and prevent film spallation and substrate smearing while polishing. I have tried to sandwich film with another Al piece, but can't get intimate contact for good support, so film is badly damaged. One suggestion was electroless plating or electroplating. Any suggestions or resources to pursue? Thanks very much.
} Date: Wed, 12 Feb 1997 09:58:05 -0600 } From: Linda Fox {lfox1-at-wpo.it.luc.edu} } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: research vs clinical work } ANSWER IN CAPS (NOT SHOUTING, JOE) } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Our lab has an opportunity to pick up some work doing clinical } specimens for $$. Currently we are strictly a research facility, but } in today's world some cash can really go a long way to justify our } existence. Can anyone advise us as to the "Chicago-style pot holes" } that may be looming unforseen on the horizon?
WHAT KIND OF SAMPLES? YOU NEED EXPERTISE IN INTERPRETING THE MATERIAL FOR WHATEVER THE PHYSICIANS WANT TO ASK (E.G., IS THERE TUMOR, IS THERE A VIRUS OR OTHER INFECTIOUS AGENT PRESENT, IS THERE AN ABNORMAL CELL STRUCTURE PRESENT?). YOU NEED SOMEONE WHOSE AUTHORITY WILL BE RESPECTED. A BOARD-CERTIFIED PATHOLOGIST IS HANDY, BUT NOT ABSOLUTELY NECESSARY.
Do we need clinical accreditation? IF YOU ARE GOING TO CHARGE PATIENTS/INSURANCE COs FOR YOUR SERVICE, YOU WILL **HAVE** TO BE CERTIFIED BY CAP AND CLIA (COLLEGE OF AMERICAN PATHOLOGISTS AND HEALTH CARE FINANCEING ADMINISTRATION CLINICAL LABORATORY IMPROVEMENT AMENDMENTS). IT IS A TEDIOUS PROCESS, BUT NOT IMPOSSIBLE. YOU HAVE TO HAVE RECORDS FOR **EVERYTHING** FROM HOW YOU MAKE UP YOUR SOLUTIONS, TO WHEN YOU CHECKED THE GROUNDING ON YOUR INSTRUMENTS, ETC.
How much should we charge for clinical work? THIN SECTIONING OR NEGATIVE STAINING? THERE IS A CPT CODE FOR ELECTRON MICROSCOPY THAT SETS THE AMOUNT MEDICARE/INS COs PAY. YOU MIGHT GET AWAY WITH SETTING IT UP AS A CONSULTING CHARGE WITHOUT THE CPT GUIDELINE. FIGURE HOW MUCH TIME AND TROUBLE YOU WILL SPEND AND THEN CALCULATE A CHARGE. NEGATIVE STAINS RUN FROM ABOUT ~$50 (HIGHLY SUBSIDIZED) TO $400, DEPENDING ON THE CONPLEXITY OF THE SPECIMEN CONCENTRATION EFFORTS; THIN SECTIONS RUN FROM ~$300-700 (NOT INCLUDING A PATHOLOGIST'S PROFESSIONAL FEE).
Any thoughts about turn around time? AGAIN: DO YOU MEAN NEGATIVE STAINING OR THIN SECTIONING??? FOR NEGATIVE STAINING 1-8 HR, DEPENDING ON WHETHER YOU HAVE TO CONCENTRATE IT, OR FOR THIN SECTIONNING 2-4 DAYS, DEPENDING ON WHETHER YOU FIND WHAT YOU'RE LOOKING FOR IMMEDIATELY OR HAVE TO HUNT FOR IT.
QC? DEFINITELY, BETTER IF FROM AN OUTSIDE LAB. Are there things we should get } straight and in writing before we begin? ABSOLUTELY. I SUGGEST YOU CONTACT A HOSPITAL EM LAB IN YOUR AREA THAT IS ALREADY CERTIFIED TO HELP YOU GET STARTED.
Thanks for the input. } } p.s. Many Thanks for the Alizarin Red stuff....it's been tried and is } working great!! } } Linda Fox } Loyola University Medical School } Maywood Illinois } lfox1-at-wpo.it.luc.edu }
Sara E. Miller, Ph. D. (DIRECTOR, ELECTRON MICROSCOPY DIAGNOSTIC VIROLOGY LABORATORY and DIRECTOR, SURGICAL PATHOLOGY ELECTRON MICROSCOPY LABORATORY) P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
A couple of things come to mind regarding your Al2O3/Al film, both of them having to do with ultramicrotomy (cutting ultrathin sections).
You may or may not need the thin sections which would be cut from the bulk material. But the nice thing about ultramicrotomy is the bulk material becomes well polished during sectioning.
Do you have access to an EM lab that has an ultramicrotome? If so, it certainly is worth a try.
You could probably adequately polish the material by first trimming a small piece of the substrate/film to a needle-like point, then clamping it in a "vise" type specimen holder for the ultramicrotome, and cutting (sectioning) in cross section at the fine point.
If the film separates from the substrate when you do this, it is possible to also embed a small sliver of the specimen in a plastic resin, and to then cut the plastic-embedded specimen. You may need to pre-treat the specimen with a silanization agent to increase the adhesion between the specimen and the resin. This assumes that surrounding the specimen with plastic is acceptable and won't interfere with whatever you need to do in the SEM.
There are lots of methods for ultramicrotomy in materials science, and the world's experts subscribe to this listserver: Tom Malis? Caroline Schooley? Phil Swab? Are you there?
Whether the material is cut naked or embedded in plastic, your friendly local ultramicrotomist should give it a try first with an old diamond knife, cutting at around 30-60 nm thickness.
A diamond knife would do a *great* job of cutting the aluminum. If your resident ultramicrotomist only cuts biological material.....Well, let's just say you'll have to do some persuading, cajoling, groveling or bribing as necessary. But polishing by ultramicrotomy works very nicely. And provided it's done properly, it will *NOT* destroy the diamond knife.
Group - While I do not keep a file on email addresses for microscopy manufacturers and suppliers, I do so for their web addresses and expect that most include their email addresses. With some 50 plus currently included, I publish the list in Microscopy Today every few months and will do so again in my upcoming issue. If our international friends would like a copy of the updated list, kindly send me an email with BOTH your email address and fax number. If I can not figure out how to send the list to you by email, I will do so by fax. And I will include how to subscribe to our publication. To manufacturers/suppliers, if I do not have your web addresses, kindly advise by return email and I will so include. Best to all - Don Grimes, Microscopy Today
} Priority: normal } Subject: Re:Degausser } From: "Scott D. Walck WL/MLBT" {walcksd-at-ml.wpafb.af.mil} } To: Microscopy ListServer {Microscopy-at-sparc5.microscopy.com} } Date: Mon, 10 Feb 97 21:56:44 -0500
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } To alleviate this try } } demagnetising by scrambling the domains in a high field strength such as in } } an old mains transformer with a cut out channeled in the metal pole piece. } } This works for us and is a cheap solution but I dare say someone will sell } } you a "degausser" which will work equally well and might look more } } acceptable to the safety officer ! Ask your physics/electronics people for } } a supplier. } } } } Regards } } } } Laurence Tetley } } } A degausser that you can buy and perhaps use for something else is a soldering } gun that has the two leads coming out to form the tip. I think that Sears } sells one like that that has a light on it. Put your tweezers slowly in and } slowly out and it will degauss them. } - -Scott Walck } A cheap tape head degauser from Tandys(they used to be about $20) will do a better job and is actially designed to degauss. It can also be used to degauss vials of grids Kerry Gascoigne ***************************************************** Kerry Gascoigne Flinders Microscope and Image Analysis Facility. Ph (08)8204-4858 Fax (08)8277-0085 ***************************************************
A small private lab is seeking an individual fluent in all phases of TEM specimen preparation. This will include all record keeping,chemistry,fixation,processing,thin/thick sectioning and darkroom work as well as accomplishing all daily associated tasks. Familiarity with computers and good laboratory practices is desirable Salary open and commensurate with experience.
Send or fax resume with cover letter to: Dr Marco Chacon Paragon Biotech,Inc Hopkins-Bayview Alpha Center 5210 Eastern Ave Baltimore,Maryland 21224 fax 410 550-2924
Dear Loren, I have had some success with thin films by glueing glass pieces about 6 mm. thick on both sides of the film, using 5 minute epoxy. After the glue is hard you can polish as usual for mounted samples, then carbon or gold coat for SEM. I have used this to analyse by EDX through a 70 micron film. Electroplating is also very effective for edge retention, but may not work on a non-conductive film. You wrote: } Dear Friends, } I have a thin (1um) al2o3/al film on an aluminum (1mm) substrate. I am } trying to mount for the cross section. I need a method to preserve the } edge and prevent film spallation and substrate smearing while polishing. I } have tried to sandwich film with another Al piece, but can't get intimate } contact for good support, so film is badly damaged. One suggestion was } electroless plating or electroplating. Any suggestions or resources } to pursue? Thanks very much. } } Loren Prentice (University of Michigan) Good luck, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
The Microscopy Vendors Database is one of the largest vendors database (company name, address, phone and fax number, e-mail and web adress, product information)and consist more than 1100 companies with about 280 web address and new keyword search engine.
http://www.kaker.com/mvd/vendors.html
Henrik
-- Henrik Kaker SEM-EDS Laboratory, Metal Ravne d.o.o., Slovenia Tel: +386-602-21-131, Fax: +386-602-20-436, E-mail: Henrik.Kaker-at-guest.arnes.si http://www2.arnes.si/guest/sgszmera1/index.html Microscopy Vendors DataBase: http://www2.arnes.si/guest/sgszmera1/vendors.html
Recently I asked for help about chemical etching of Si and received many excellent advises. Thank you for your attention and help.
First of all, why do we use chemical etching? 1)Variouse jet technigues required spesial devises that allow to operate with HF base solutes, unfortunately, we have no such one. 2)Ion etching is very powerful method, but we have found very small radiation damage produced during ion etching of silicon.
We have solve the problem of pitting during Si [111] chemical etching. At the end the best solute is: HF: HNO_3 : ( CH_3COOH + I_2 cryst.) 3 : 9 : 8 2.5g cryst. I_2 per 1100ml CH_3COOH (is a good job to dissolve I_2 in a hot acid)
Average etching rate of virgin solute at room temperature is less than 3 micron/min from one side.
In addition the rest of the message contains all the responses deal with the chemical etching of Silicon from Microscopy Society.
Sorry if took so long to respond. Kirill Prikhodko. Russian Research Center "Kurchatov Institute" Moscow E-mail: kirill-at-nw.oirtorm.net.kiae.su
With regard to Kirill Prikhodko's request, a recommended procedure for preparing Si is by chemical jet etching. The solution used is typically HF based. There are many which are quite sufficient. The following are a few which have been proven satisfactory:
1.) 90% Nitric acid 10% HF 2.) 5 parts Nitric acid, 3 parts Acetic acid, 3 parts HF. Polish at room temperature. 3.) 10.5 grams Potassium Permanganate, 300ml HF, 30ml De-ionized Water. Polish at -20 to -30 degrees C with a high jetting speed.
Depending on the orientation of the Si, the percentage of the acids may need to be altered.
These solutions and conditions have provided excellent results when utilizing the twin-jet electropolishing technique which simultaneously thins both specimen surfaces. If it is desired to back-thin the Si, one side should be protected with Beeswax.
For the electrolytic polishing of metals it is recommended to have both the specimen and the jets submerged in the electrolyte by approximately 3- 4mm. When chemical etching of Si, it is recommended that the specimen be above the chemical solution level. The jet position in relation to the specimen can be varied to provide the necessary configuration of the dimple produced by the chemical etching process.
Kind regards for a Happy Holiday Season,
Paul Fischione email: Paul.Fischione-at-internetmci.com
E.A. Fischione Instrument, Inc. is the manufacturer of the Automatic Twin- Jet Electropolisher.
====================================================================== Froim: Tan-Chen Lee
Reply to: RE} TEM: Si chemical etching
It is not the ratio of acids which caused problem. I used to do chemical etching in Taiwan. It went fine. However, the same recipe resulted in pitting on Si(100) but not on Si(111) when I tried it in New York. It seemed that preferential etching happened. Even when I did it in clean room or change recipes, it did not solve the problem. I suspect the possible reasons are the contents of the acids (concentration, contaminants, etc.), temperature, sample holders (Teflon versus glasses??), wax, etc. You may only need to change the vendors of the chemicals. I would also like to know the real cause though I do not do chemical ethcing any more.
Materials Characterization Lab Motorola, Inc. email: tan-chen_lee-at-mesaqm.sps.mot.com
} Dear all } } Can some one please send me website for downloading the shareware } version of Electron Flight Simmulater.
Dear Stephan,
You can download the shareware version 3.1 for Windows of Electron Flight Simulator from website of Small World Co., http://members.aol.com/smworld100/efs.htm
Vladimir Oleshko *********************************************************** V.P. Oleshko, Ph.D e-mail:oleshko-at-uia.ua.ac.be Micro-and Trace Analysis Centre Tel.:+32-3-820.23.64 Chemistry Department FAX :+32-3-820.23.76 University of Antwerp (UIA) B-2610 Belgium ***********************************************************
Either Buhler or Struers (or both?), metallographic suppliers, sell an electroless Ni plating solution which may help. The host material does not have to be conductive, but some materials plate better than others. Do check that the solution will not attack your sample and note that the plate is really a nickel/phosphorus compound. Woody ______________________________ Reply Separator _________________________________
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Friends, I have a thin (1um) al2o3/al film on an aluminum (1mm) substrate. I am trying to mount for the cross section. I need a method to preserve the edge and prevent film spallation and substrate smearing while polishing. I have tried to sandwich film with another Al piece, but can't get intimate contact
for good support, so film is badly damaged. One suggestion was electroless plating or electroplating. Any suggestions or resources to pursue? Thanks very much.
A postdoctoral position is available for a candidate with expertese in transmission electron microscopy to work on a project involving intracellular trafficking. Some expertese in immunohistochemistry/cytochemistry would be desirable. For further information please contact Dr. Henry Hoff, Department of Cell Biology, Cleveland Clinic Foundation, 9500 Euclid Ave., Cleveland, Ohio 44195 Tel: (216) 444-2248 or e-mail: {hoffh-at-cesmtp.ccf.org }
I am a student of a doctoral program in Biological Sciences of the Universidad Autonoma de Nuevo Leon (Autonomous University of Nuevo Leon) at Monterrey, Mexico. I am interested in a Visiting Scientist Program of an institution, in order to participate in a research, as a doctoral thesis. I have a Bachelor degree in Biological Sciences of the Faculty of Biological Sciences, and a Master degree in Morphology of th Faculty of Medicine, both of them of the university I mentioned. My main experience is in histotechnology for light and transmision electron microscopy, ultraestructure of mitochondria, cell culture and genetic of insects. I send this message to this forum to ask if anyone could send me information or some tips that could be of help to me in contacting an opportunity. My data are: Ricardo Acosta Nueva Independencia 308 Colonia Independencia 64720 Monterrey, Nuevo Leon Mexico.
Does anyone have any experience with and/or opinions/comments about the Linker software, written by a Finnish company called PICOMEGA, for the manipulation of images exported to DOS from older LINK/Oxford computers with DEMON operating systems? My interest arises because we have a LINK QX2000 system which can produce reasonable element maps, BSE images, and linescan images onscreen, but is extremely limited in its ability to produce hard copy.
An email or fax address of the company would also come in handy (or, as last resort, their phone number)
thanks
Ritchie
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
I am looking for a materials scientist with extensive TEM experience to fill the following position at the IBM Almaden Research Center. Please contact me directly.
A one-year position is available at the IBM Almaden Research Center to perform TEM studies on magnetic recording materials. The work would involve extensive collaboration with the groups at Almaden and in the IBM Storage Systems Division that are developing next- generation heads and disks. Candidates should have an M.S. or Ph.D. degree in materials science (or a related field), extensive TEM experience, and preferably some background in magnetic recording.
The position is available immediately. Funding beyond the first year is probable, but cannot be guaranteed at this time.
Applicants should submit their resumes to:
Robby Beyers K19/D1 IBM Almaden Research Center 650 Harry Road San Jose, CA 95120-6099
Following my recent posting about lab services, I had a number of requests for details. There is a lot of data, so it would not be appropriate to post it to the Microscopy List. For those who contacted me directly, I have attached, as ASCII text files, both the USA and UK list which we maintain.
Currently, we do not have a wider European list, since this has not seemed appropriate for a paper publication. However, we are in the process of establishing a web site, which will include all the data we currently hold, and will be extended in the future to cover the rest of Europe.
When our web site is up, I'll post details to the Microscopy list.
Regards,
--------------------------------------------------------------- Dr L. P. Stoter Technical Editor, MICROSCOPY & ANALYSIS Technesis 17, Rocks Park Road email: LPS-at-teknesis.demon.co.uk Uckfield, E. Sussex Phone: +44 (0)1825 766911 TN22 2AT Fax: +44 (0)1825 766911 United Kingdom ---------------------------------------------------------------
Does anybody have any advice on aligning a microscope? I have an afocal pair to relay the exit pupil and a tube lens with an infinity corrected objective and would like to know how to align them all as well at the eppi illumionation path very precisely since I am doing subresolution (1% of diffractuion limit) measurements in phase contrast mode.
} } Does anyone have any experience with and/or opinions/comments about } the Linker software, written by a Finnish company called PICOMEGA, } for the manipulation of images exported to DOS from older LINK/Oxford } computers with DEMON operating systems? } My interest arises because we have a LINK QX2000 system which can } produce reasonable element maps, BSE images, and linescan images } onscreen, but is extremely limited in its ability to produce hard } copy.
I can't help with the direct question, but I have written software to do the following:
Read AN10000/QX2000/eX/L disks on a PC, and copy their contents to the PC hard disk.
Read the spectrum files (-.SP) and convert them to ASCII spreadsheet format
Read studies (-.SY) and extract the individual images and save them as TIFF files
Format Demon/Demon Plus/Genie disks on the PC
These programs only run in plain vanilla DOS (not DOS within Windows) but (for us, at least,) are a whole lot more convenient than using the LINK computers. They are free to anybody who asks. I will set up an FTP site with them - I will post the address here when it is done.
Tony Garratt-Reed }
**************************************************** **************************************************** ** ** ** Anthony J. Garratt-Reed ** ** Room 13-1027 ** ** Center for Materials Science and Engineering ** ** Massachusetts Institute of Technology ** ** 77 Massachusetts Avenue ** ** Cambridge, Massachsetts 02139-4307 ** ** U. S. A. ** ** ** ** Phone: 617-253-4622 ** ** Fax: 617-258-5286 or 617-258-6478 ** ** ** **************************************************** ****************************************************
Dear Friends, Thanks for all of your suggestions for mounting and polishing a thin film for cross-section examination. The advice fell generally into several areas. One group of suggestions involved encapsulating the film/substrate, either with electroless Nickel plating, a glass support, a hard resin, and the like. A number of people also suggested microtoming either the naked sample or embedding the sample in resin and then sectioning to obtain a good polish in the process. Microtoming looks like the easiest route for now, since we have the facilities here at UM. Thanks again for the help and if anyone has questions, I could point you to some of the sources of my advice. Best regards, Loren Prentice
Our lab has been asked to characterize lyophilized human lung tissue by SEM and XRF analysis. Not being pathologists and having no prior experience with biological tissue samples, we are concerned about potential health risks from handling this material. The samples were collected and lyophilized at least 20 years ago and have been in storage all this time. Sample preparation for the XRF analysis will require pulverizing the material with mortar and pestle and depositing this fine dust onto filter subtrates for analysis with the potential for exposure to or inhalation of the dust. Our safety officer doesn't know whether any viruses or bacteria could still be viable in any of these samples, and is not sure what level of safety precautions are required: e.g., Should the work be done in a hood certified for biohazard work or is this overkill? Should the lab technician be inoculated against hepatitis B? Moon suits and hazard pay? (I'm joking, but maybe I shouldn't be). The histories of the tissue donors are available if that would be a determining factor but I don't think many died of an infectious disease.
Thanks for any comments and suggestions.
Bob Willis ManTech Environmental email: Willis.robert-at-epamail.epa.gov
If I were you, I would take every precaution that you would with fresh tissue. If stored properly , lyophilized microbes can survive a long time. When I was a postdoc we got live cultures from lyophilized samples that were under vacuum and stored at 4 degrees C for over 40 years. This was not an exception but rather the rule. So if these samples have not been fixed or other denatured or sterilized, I would be careful. } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } ..
At 12:04 PM 2/14/97 -0500, you wrote:
} Our lab has been asked to characterize lyophilized human lung tissue by ******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
John Winter is visiting my EPMA facility and has asked about PC platformed hardware and software which would interface with an existing SEM/EDX detector. All of my info is several years old ... and I think John would prefer anyway to hear from satisfied users as well as vendors. John has not expressed any preference for PC vs Macintosh. Replies should be preferably sent to John {winterj-at-whitman.edu} or myself ... TIA ...
cheers, shAf
{\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo ZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We have an immediate opening for a jr. level scanning electron microscopist at our Bedford Ma. Facility. Please send all correspondence to the address found below the job posting.
Millipore Corporation's Scanning Electron Microscopy Lab is seeking an individual to assist in the evaluation of customer complaints, perform failure analysis and/or troubleshooting on existing products and characterize experimental polymer membranes structures through the use of scanning electron microscopy, light microscopy, energy dispersive spectroscopy and other imaging techniques. You will be responsible for the generation, interpretation and written/verbal communication of both routine and non-routine micrographic and spectral results to both in-house clients and external Millipore customers. You will also be responsible for the maintenance of lab equipment (SEM's, LM's, PC's, AV equipment etc.). You may also be asked to conduct customer tours and present overviews of lab capabilities to sales training groups. REQUIREMENTS: BS/BA degree in a technical field or equivalent and a minimum of 2 years experience in scanning electron microscopy with a focus on materials, preferably in a support lab environment. Must be extremely flexible and able to work in a high pressure environment. Must possess a fundamental understanding of electron beam interactions, electron optics and vacuum systems. Experience with high pressure SEMs a plus. Must also have a working knowledge of personal computing hardware/software systems. Strong written and verbal communication skills are essential.
To apply mail your resume to:
Employment Manager Millipore Corporation 80 Ashby Rd. Bedford, MA 01730 OR Send your resume via Email to: careers-at-millipore.com. Please, NO FAXES. We Scan all resumes into a Database and Faxed copies do not scan well.
What are some of the ways you can get rid of waste osmium? We already add it to excess corn oil but our safety people say it's still toxic. I thought I read once a technique that neutralizes it allowing it to be flushed down the sink or at the minimum placed in with regular disposal products? Our radiation people would like to see a similar thing with my uranyl acetate. Thanks for any advice.
What are some of the ways you can get rid of waste osmium? We already add it to excess corn oil but our safety people say it's still toxic. I thought I read once a technique that neutralizes it allowing it to be flushed down the sink or at the minimum placed in with regular disposal products? Our radiation people would like to see a similar thing with my uranyl acetate. Thanks for any advice.
What are some of the ways you can get rid of waste osmium? We already add it to excess corn oil but our safety people say it's still toxic. I thought I read once a technique that neutralizes it allowing it to be flushed down the sink or at a minimum placed in with regular disposal products? Our radiation people would like to see a similar thing with my uranyl acetate. Thanks for any advice.
The School of Materials Science and Engineering at Georgia Institute of Technology is looking for a side-entry TEM (100 or 120 kV) for conventional research and teaching. The TEM must have a high degree double tilting specimen stage. Please reply to this e-mail if you have such a microscope in your lab for sale. Thanks.
} Date: Fri, 14 Feb 1997 12:04:47 -0500 } From: ROBERT WILLIS {WILLIS.ROBERT-at-EPAMAIL.EPA.GOV} } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Health risks of lyophilized lung tissue? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Our lab has been asked to characterize lyophilized human lung tissue by } SEM and XRF analysis. Not being pathologists and having no prior } experience with biological tissue samples, we are concerned about } potential health risks from handling this material. The samples were } collected and lyophilized at least 20 years ago and have been in storage } all this time. Sample preparation for the XRF analysis will require } pulverizing the material with mortar and pestle and depositing this fine } dust onto filter subtrates for analysis with the potential for exposure to } or inhalation of the dust. Our safety officer doesn't know whether any } viruses or bacteria could still be viable in any of these samples,
POSSIBLY
and is not sure what level of safety precautions are required: e.g., Should the } work be done in a hood certified for biohazard work or is this overkill?
DEFINITELY. NOT OVERKILL.
} Should the lab technician be inoculated against hepatitis B?
JUST DON'T STAB YOURSELF WITH CONTAMINATED FORCEPS.
Moon suits } and hazard pay? (I'm joking, but maybe I shouldn't be). The histories of } the tissue donors are available if that would be a determining factor but I } don't think many died of an infectious disease. } I'D WORRY MORE ABOUT TB THAN HEPATITIS. TB IS VERY INFECTIOUS IN AEROSOLS AND DUST. I'D MAKE SURE NONE OF THE DUST ESCAPES, OR FIX IT SOMEHOW, IF YOU CAN KEEP FROM CONTAMINATING YOUR ANALYSIS WITH SOMETHING THAT WOULD RUIN YOUR TEST. HOW ABOUT OSMIUM VAPOR??? JUST DON'T TAKE ANY CHANCES. REGULATIONS NOW REQUIRE THAT YOU TREAT ALL TISSUES AND BODLIY FLUIDS AS THOUGH THEY MAY BE INFECTIOUS.
} Thanks for any comments and suggestions. } } Bob Willis } ManTech Environmental } email: Willis.robert-at-epamail.epa.gov }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Space is getting tight and we are thinking of moving some computers into the same room as our vacuum evaporator. Although we have an oil mist filter on the VE, we get some smell from the rotary pump when roughing out the bell jar. Any suggestions on how to eliminate this problem, the computer users have objected to the smell and possible health concerns.
Are some filters better than others? How realistic is it to expect a filter to eliminate all odor? I have considered venting the pumps to another room and putting a big industrial filter there, but am worried about oil condensing out in the vent pipe on its way to the filter. The campus facilities folks are reluctant to vent the pumps into the building exhaust because it will involve rebalancing the whole building etc. But they might be up for putting in a vent to another room if we can figure out how to do it without creating other problems.
Any ideas?
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
Dear Jon, There are several ways of dealing with rotary pump vapours. One pump salesman told me that the best filter was a giant, movie-house bag of popcorn! Great oil absorber and cheap to replace. I have vented most of my pumps out the window or into the fume cupboard. I'm very surprised that the building people were concerned about the building exhaust, since the vent pipe is so small (one inch or less) and there is almost no movement of air through the vent pipe, and that only for a few seconds until a bit of vacuum is established. If you lead a long pipe into another room, there is no harm in the vapours condensing in the pipe, that eliminates some of the problem. The computer users are correct, oil vapours are a health hazard if inhaled. You wrote: } Space is getting tight and we are thinking of moving some computers into } the same room as our vacuum evaporator. Although we have an oil mist filter } on the VE, we get some smell from the rotary pump when roughing out the } bell jar. Any suggestions on how to eliminate this problem, the computer } users have objected to the smell and possible health concerns. } } Are some filters better than others? How realistic is it to expect a filter } to eliminate all odor? I have considered venting the pumps to another room } and putting a big industrial filter there, but am worried about oil } condensing out in the vent pipe on its way to the filter. The campus } facilities folks are reluctant to vent the pumps into the building exhaust } because it will involve rebalancing the whole building etc. But they might } be up for putting in a vent to another room if we can figure out how to do } it without creating other problems. } } Any ideas? } } } } Jonathan Krupp } Microscopy and Imaging Lab } University of California } Santa Cruz, CA 95064 } (408) 459-2477 } FAX (408) 429-0146 } jmkrupp-at-cats.ucsc.edu } } } } Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
} Our lab has been asked to characterize lyophilized human lung tissue by } SEM and XRF analysis. Not being pathologists and having no prior
snips
} Should the lab technician be inoculated against hepatitis B? Moon suits } and hazard pay? (I'm joking, but maybe I shouldn't be). The histories of } the tissue donors are available if that would be a determining factor but I } don't think many died of an infectious disease. } } Thanks for any comments and suggestions. } } Bob Willis } ManTech Environmental } email: Willis.robert-at-epamail.epa.gov
This is not my area but I've done quite a lot of EM on viruses for others. Several have commented to me that putting a virus in the electron beam, which has similarities to putting it at the centre of a small nuclear explosion, is probably the only sure way to kill a virus. And until that point, they always regard a virus as 'alive', whatever chemicals it may have gone through.
} Space is getting tight and we are thinking of moving some computers into } the same room as our vacuum evaporator. Although we have an oil mist filter } on the VE, we get some smell from the rotary pump when roughing out the } bell jar. Any suggestions on how to eliminate this problem, the computer } users have objected to the smell and possible health concerns. } } Are some filters better than others? How realistic is it to expect a filter } to eliminate all odor? I have considered venting the pumps to another room } and putting a big industrial filter there, but am worried about oil } condensing out in the vent pipe on its way to the filter. The campus } facilities folks are reluctant to vent the pumps into the building exhaust } because it will involve rebalancing the whole building etc. But they might } be up for putting in a vent to another room if we can figure out how to do } it without creating other problems. } } Any ideas? } } } } Jonathan Krupp } Microscopy and Imaging Lab } University of California } Santa Cruz, CA 95064 } (408) 459-2477 } FAX (408) 429-0146 } jmkrupp-at-cats.ucsc.edu
If you can smell oil, then oil is present!
When new, I feel that some oil mist filters are quite effective, but over time I'm sure their efficiency drops significantly. Personally, in any installation where a rotary pump is frequently moving large volumes, such as evaporators and SEMs, I would recommend venting the rotary pump to outside the building.
We too sometimes have Ni grids moving around tweezers tips. To find if tweezers are magnetised we use a small compass - a 1 inch toy borrowed to some kid - that is sensitive enough to diagnose the magnetisation. To solve the problem, and as it was proposed in another message, the use of a demagnetise is really most useful.That we use to demagnetise the tweezer and not the grids. We use a small one, priced about USD 150, and bought from Agar Scientific Ld, Essex CM24 8D4, England (the FAX was - some time ago - the (0279) 815106) It is a very small and very effective equipment that can be used for demagnetise tweezers, screw drivers and other small metallic tools.
{DT} We too sometimes have Ni grids moving around tweezers tips. To find if tweezers are magnetised we use a small compass - a 1 inch toy borrowed to some kid - that is sensitive enough to diagnose the magnetisation. To solve the problem, and as it was proposed in another message, the use of a demagnetise is really most useful.That we use to demagnetise the tweezer and not the grids. We use a small one, priced about USD 150, and bought from Agar Scientific Ld, Essex CM24 8D4, England (the FAX was - some time ago - the (0279) 815106) It is a very small and very effective equipment that can be used for demagnetise tweezers, screw drivers and other small metallic tools. {/DT}
Following my message yesterday, I have put my collection of utilities to convert and read Link format files (AN1000, QX2000 and eX/L) on IMAGES.MIT.EDU
You can FTP to there (anonymous works, it wants your e-mail address as a password) and have a look at them. Have fun!
Let me know if you have problems.
Tony Garratt-Reed
**************************************************** **************************************************** ** ** ** Anthony J. Garratt-Reed ** ** Room 13-1027 ** ** Center for Materials Science and Engineering ** ** Massachusetts Institute of Technology ** ** 77 Massachusetts Avenue ** ** Cambridge, Massachsetts 02139-4307 ** ** U. S. A. ** ** ** ** Phone: 617-253-4622 ** ** Fax: 617-258-5286 or 617-258-6478 ** ** ** **************************************************** ****************************************************
} Date: Sat, 15 Feb 1997 09:22:36 +0000 } From: Larry Stoter {LPS-at-teknesis.demon.co.uk} } To: ROBERT WILLIS {WILLIS.ROBERT-at-EPAMAIL.EPA.GOV} , } Microscopy-at-Sparc5.Microscopy.Com } Subject: Re: Health risks of lyophilized lung tissue? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Our lab has been asked to characterize lyophilized human lung tissue by } } SEM and XRF analysis. Not being pathologists and having no prior } } snips } } } Should the lab technician be inoculated against hepatitis B? Moon suits } } and hazard pay? (I'm joking, but maybe I shouldn't be). The histories of } } the tissue donors are available if that would be a determining factor but I } } don't think many died of an infectious disease. } } } } Thanks for any comments and suggestions. } } } } Bob Willis } } ManTech Environmental } } email: Willis.robert-at-epamail.epa.gov } } This is not my area but I've done quite a lot of EM on viruses for others. } Several have commented to me that putting a virus in the electron beam, } which has similarities to putting it at the centre of a small nuclear } explosion, is probably the only sure way to kill a virus. And until that } point, they always regard a virus as 'alive', whatever chemicals it may } have gone through. } } Regards, } Larry Stoter } Larry and Bob,
There are many ways of killing viruses, besides electron beams; however, I wouldn't assume that the electron beam hits every nm of space on the sample, and hence, kills everything that went into the scope!!} Furthermore, the dust from grinding specimen flying around while you're preparing it for EM could be infectious. See my earlier comment on TB.
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
ROBERT WILLIS {WILLIS.ROBERT-at-EPAMAIL.EPA.GOV} wrote:
} Our lab has been asked to characterize lyophilized human lung tissue by } SEM and XRF analysis. Not being pathologists and having no prior } experience with biological tissue samples, we are concerned about } potential health risks from handling this material. The samples were } collected and lyophilized at least 20 years ago and have been in storage } all this time.
Ah nostalgia! 20 years ago I was starting my Path residency. Pathogens in the specimens may very well still be viable. At least you don't have to worry too much about HIV- but I wouldn't have been worried about that anyways- it's not known to be transmissible by inhalation. My major concern would be tuberculosis. Most of the bacteria and viruses that might lurk in old lungs are unlikely to cause serious disease in a person with an intact immune system. TB can cause a serious infection despite a good immune system and infection can be established with a very small dose.
} Sample preparation for the XRF analysis will require } pulverizing the material with mortar and pestle and depositing this fine } dust onto filter subtrates for analysis with the potential for exposure to } or inhalation of the dust.
Now I'm really concerned about TB. The pulverization is a perfect way to get aerosols into your lungs.
} Our safety officer doesn't know whether any } viruses or bacteria could still be viable in any of these samples, and is } not sure what level of safety precautions are required: e.g., Should the } work be done in a hood certified for biohazard work or is this overkill? } Should the lab technician be inoculated against hepatitis B? Moon suits } and hazard pay? (I'm joking, but maybe I shouldn't be).
Doing the work in a hood is not a bad idea. You could probably get away with having everyone in the room wear a respirator with a filter fine enough to filter out TB ("N95" respirator). These are available as powered positive air pressure units or as disposable non-powered units. A surgical face mask would not be sufficient. Hepatitis B vaccination shouldn't be necessary, it's another bug that probably isn't transmitted by inhalation, but the vaccine is low risk and inexpensive so why not?
} The histories of } the tissue donors are available if that would be a determining factor but I } don't think many died of an infectious disease.
You might want to look at occupational histories. Some groups, e.g. miners, had higher incidence of TB.
I hope this helps.
Leon
-- Leon A. Metlay, M.D.,Associate Professor of Pathology and Laboratory Medicine University of Rochester Medical Center Phone: (716) 275-5691 P.O. Box 626 Fax: (716) 273-1027 Rochester, NY 14642 lmetlay-at-acu.pathology.rochester.edu http://www.urmc.rochester.edu/smd/pathres/URPLM.html "Most ass drivers are evil, most camel drivers are decent, most sailors are saintly, the best among physicians is going to Gehenna, and the best of butchers is a partner of Amalek" -R. Judah, in Mish. Kidd. 4:14
If at all possible try to vent your RP into a fume hood or outside. You can also double filter the exhaust by making a 3-4" dia PVC sealed tube about 10-12" long with screw cap plugs on each end. I've done this in the past with good results.
You have to drill a hole in the top and botton and thread in a pipe nipple the same size as the pump outlet at the bottom of the cylinder. Adapt this to fit the threads ofa Balston disposable vapor filter which will sit on the inside of the cylinder. Drill another hole in the top cap and tap it to fit another Balston filter. I can fax you a drawing if you are interested. Just emailme your FAX #.
cheers
Ed Basgall, PhD Penn State Univ Dept of Chem University Park, PA 16801
I am a pathology resident at Johns Hopkins. We currently use the Roche(now autocyte) digital camera to capture high resolution/publication quality photomicroscopic images of pathology slide specimens. My question is what is your experience with other vendors? Are there high quality cameras out there that can easily be linked to a Zeiss or other microscope? We have learned a bit about pixera and kodak cameras, but have been less than fully impressed with the technical help we have received.
Angelo M. De Marzo MD/PhD ademarz-at-welchlink.welch.jhu.edu
To: Microscopy ListSer {Microscopy-at-MSA.Microscopy.Com} *** Reply to note of 02/15/97 02:27
Bob Chiovetti, I'm doing the "Microscopy 101" column in Microscopy Today, and I'd like to use your answer in the column, or an edited version of it. Please let me know. Thanks! Phil
&&& Illigitimi non carborundum &&&&&&&& Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217)244-3145 days (217)355-3145 evenings oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
Loren Prentice, I'm doing the "Microscopy 101" column in Microscopy Today, and I'd like to use your question in the column, or an edited version of it. Please let me know. Thanks! Phil
****be famous! send in a tech tip or question*** Philip Oshel Technical Editor Microscopy Today Station A PO Box 5037 Champaign, IL 61825-5037
} From: J.F.Moura Nunes {vmnunes-at-mail.telepac.pt} } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Re: TEM - Ni grids & magnetized tweezers } Date: Sunday, 16 February 1997 5:29 } We too sometimes have Ni grids moving around tweezers tips. To find if tweezers are magnetised we use a small compass - . . . . . cut J.F.Moura Nunes {vmnunes-at-mail.telepac.pt} **************************** The most commonly used steel in "EM" tweezers is "Inox" which in the Dumont range indicates surgical steel- which is magnetic. When using nickel grids or other materials which may be subject to magnetism it's simplest to use tweezers made from non-magnetic steel. Dumont use two: Dumoxel and Dumostar. The latter is by far the best steel for fine tweezers, it is highly acid, alkali and seawater resistant and has terrific hardness and elasticity properties. They are the most expensive, but in the hands of a skilled operator they are the most economic tweezers too. Read up about that relatively new steel. Note: ProSciTech supplies Dumont (and other) tweezers. To save other suppliers the hassle, please note that all major EM suppliers stock tweezers too.
Jim Darley ProSciTech Microscopy Supplies & Accessories PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, links, MSDS ****************** http://www.proscitech.com.au
I am looking to purchase a CCD camera to obtain images of liquid samples flowing through a slit die, to study birefringance and flow patterns. I am currently considering a 3-chip colour CCD as this will apparently give us better resolution than out current single chip colour CCD.
The main issues with our experiments is that we have good spacial resolution and can see fluid strucuture at high flow rates, colour is also reasonably important. So in an ideal case we want a high speed, high resolution colour camera, however we will also want to use a zoom lens with the camera and this is apparently problematic with 3-chip CCD's.
Larry Storter posted a message as follows: ================================================= Following my recent posting about lab services, I had a number of requests for details. There is a lot of data, so it would not be appropriate to post it to the Microscopy List. For those who contacted me directly, I have attached, as ASCII text files, both the USA and UK list which we maintain. Currently, we do not have a wider European list, since this has not seemed appropriate for a paper publication. However, we are in the process of establishing a web site, which will include all the data we currently hold, and will be extended in the future to cover the rest of Europe. ================================================= From the perspective of one who has ended up on any number of such listings over the years, I would like to suggest that any such listing would be of far greater value to prospective clients (e.g. users of such laboratory services) if information about a laboratory's accreditations and certifications accompanied such listings. For example, some laboratories are accredited to preform TEM air samples for asbestos, others are accredited to the standard of ISO Guide 25, something of great importance to any organization supporting an ISO 9000 type certification.
The best way to ensure the greatest possible accuracy, if not also honesty, in the way one represents their laboratory's credentials, is to require that when the information is submitted, it be accompanied with some kind of signed and notarized certification, attesting to the accuracy of the information being submitted.
With the growing attention being paid to quality, quality systems generally, and ISO 9000 certifications systems specifically, any such listing of laboratories at the very least should indicate those laboratories currently accredited to the standard of ISO Guide 25. The main organization doing such accrediting in North America is A2LA or American Association for Laboratory Accreditation. You can find their website at {http://members. aol.com/a2la/index.html} . In other countries, other agencies do the accrediting.
There is a huge difference between an accredited laboratory being run as a legitimate laboratory business entity as indicated above vs. the all-too- often situation when someone "runs" commercial samples for commercial clients out of a university or some other laboratory when no one is looking . It is my belief that any listing of laboratories doing EM work as a commercial service should in some way differentiate between such situations.
Disclaimer: Structure Probe, Inc. is an independent analytical laboratory offering electron microscopy services for clients, for a fee, since 1970.
Chuck
====================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ =====================================================
Can anyone tell me how to oxidize surface of a tantalum disk in order to perform the alignment of the guns in an ion mill. It must be something simple but I do not know it.
Thanks,
Yves MANIETTE Universitat de Barcelona Serveis Cientifico Tecnics Unitat ESCA TEM Carrer Lluis Sole i Sabaris, 1-3 E-08028 BARCELONA ESPANYA
I have a reference that may help you. Apparently, a 2% solution of OS4 can be neutralized by the very technique that you are using (i.e. corn oil). You should use twice as much corn oil as OS4. The reference is:
Cooper, K. Neutralization of OS4 in case of accidental spillage and for disposal. Bulletin of the Microscopical Society of Canada. 1988. 8:24-28.
Regards,
-Bob ********************************** Bob Citron Chiron Vision 555 W. Arrow Hwy Claremont, CA 91711 USA ph: (909)399-1311 email: Bob_Citron-at-cc.chiron.com **********************************
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
What are some of the ways you can get rid of waste osmium? We already add it to excess corn oil but our safety people say it's still toxic. I thought I read once a technique that neutralizes it allowing it to be flushed down the sink or at the minimum placed in with regular disposal products? Our radiation people would like to see a similar thing with my uranyl acetate. Thanks for any advice.
Someone, a week or so ago (I deleted the message), asked for info about staining of silicone in tissue. I happened to come across the following reference which may be of interest: Raso D et al. 1994. Light microscopy techniques for the demonstration of silicone gel. Arch. Pathol. Lab. Med. 118: 984-987. There also was an accompanying editorial in the same issue by Roggli et al.
In message {199702150110.RAA20593-at-cats-po-1} Jon Krupp writes: } snip } Although we have an oil mist filter } on the VE, we get some smell from the rotary pump when roughing out the } bell jar. Any suggestions on how to eliminate this problem, the computer } users have objected to the smell and possible health concerns. } } Are some filters better than others? How realistic is it to expect a filter } to eliminate all odor? } snip } Any ideas? } } Jonathan Krupp } Microscopy and Imaging Lab } University of California } Santa Cruz, CA 95064 } (408) 459-2477 } FAX (408) 429-0146 } jmkrupp-at-cats.ucsc.edu
Jonathan,
I use Balston, Inc. filter Type 9955-12, Grade 371H on my vacuum evaporator mechanical pump, two Sargent-Welch mechanical pumps, and a Marvac Z-30 direct drive pump I use for pumping my vacuum desiccator for TEM film. They have a threaded mount that fits many mechanical pump oil mist eliminator ports directly. In the case of the Sargent-Welch, I ordered an adaptor from big supplier of those pumps. like Fisher, etc., or adapt your own.
The metal oil mist "eliminators" that come with many mechanical pumps are often totally useless. This Balston filter eliminates 99% of oil vapor mist, plus they have a small tube that sticks out the bottom side for oil to drain out of. I put a short length of plastic lab tubing from it to a small 50cc bottle to collect filtered oil and eventually just put it back into the pump once per year. They are inexpensive.
Balston is in Lexington, MA. Try these filters. Good luck!
--
Gib Ahlstrand, MMS Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
My understanding is that vegetable oil reduces the osmium teroxide to less reactive oxides and elemental osmium. These tend to be far less harzardous but are still toxic, as are many heavy metals. So although you reduce the hazard, you do not eliminate it. I would assume that reduced osmium would be in the same class with lead paint and photographic silver waste. I am not really sure that there are any standards out there for disposal. AT least my safety people couldn't find them. At one time they wanted to take my blackened refrigerator, seal it in a box with vermiculite and transport to some super toxic dump site in Georgia. I refused to let them do it until they could show me the regulations on reduced osmium tetroxide and they could find none. In the meantime I painted the inside of the frig. Our safety people take our reduced osmium waste and do something with it.
Our uranyl acetate and lead citrate are combined and precipitaed with phosphate. The liquid is decanted adn sent down the drain. The solids are then turned over to safety people. This, at least, reduces the volume of waste. ******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Please forward information to me, if you have, about paper developer processors users have in their lab. I need to replace our Kodak paper developer. At least one hundred 8x10 sheets of resing coated paper each day will be processed. Price and repair history will be much appreciated!
X-Authentication-Warning: unixg.ubc.ca: lesley owned process doing -bs
Wearing a respirator is a lot better than nothing. But surely, pulverising the tissue out on the open bench will cause fine particles to fly all over the lab with every air current, contaminating everything and everybody in the lab. I would use a biohazard hood.
Lesley Weston Oral Biology University of British Columbia Vancouver, B.C., Canada
On Sat, 15 Feb 1997, Leon A. Metlay wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } ROBERT WILLIS {WILLIS.ROBERT-at-EPAMAIL.EPA.GOV} wrote: } } } Our lab has been asked to characterize lyophilized human lung tissue by } } SEM and XRF analysis. Not being pathologists and having no prior } } experience with biological tissue samples, we are concerned about } } potential health risks from handling this material. The samples were } } collected and lyophilized at least 20 years ago and have been in storage } } all this time. } } Ah nostalgia! 20 years ago I was starting my Path residency. Pathogens in } the specimens may very well still be viable. At least you don't have to } worry too much about HIV- but I wouldn't have been worried about that } anyways- it's not known to be transmissible by inhalation. My major concern } would be tuberculosis. Most of the bacteria and viruses that might lurk in } old lungs are unlikely to cause serious disease in a person with an intact } immune system. TB can cause a serious infection despite a good immune } system and infection can be established with a very small dose. } } } Sample preparation for the XRF analysis will require } } pulverizing the material with mortar and pestle and depositing this fine } } dust onto filter subtrates for analysis with the potential for exposure to } } or inhalation of the dust. } } Now I'm really concerned about TB. The pulverization is a perfect way to } get aerosols into your lungs. } } } Our safety officer doesn't know whether any } } viruses or bacteria could still be viable in any of these samples, and is } } not sure what level of safety precautions are required: e.g., Should the } } work be done in a hood certified for biohazard work or is this overkill? } } Should the lab technician be inoculated against hepatitis B? Moon suits } } and hazard pay? (I'm joking, but maybe I shouldn't be). } } Doing the work in a hood is not a bad idea. You could probably get away } with having everyone in the room wear a respirator with a filter fine } enough to filter out TB ("N95" respirator). These are available as powered } positive air pressure units or as disposable non-powered units. A surgical } face mask would not be sufficient. Hepatitis B vaccination shouldn't be } necessary, it's another bug that probably isn't transmitted by inhalation, } but the vaccine is low risk and inexpensive so why not? } } } The histories of } } the tissue donors are available if that would be a determining factor but I } } don't think many died of an infectious disease. } } You might want to look at occupational histories. Some groups, e.g. miners, } had higher incidence of TB. } } I hope this helps. } } Leon } } } -- } Leon A. Metlay, M.D.,Associate Professor of Pathology and Laboratory Medicine } University of Rochester Medical Center Phone: (716) 275-5691 } P.O. Box 626 Fax: (716) 273-1027 } Rochester, NY 14642 lmetlay-at-acu.pathology.rochester.edu } http://www.urmc.rochester.edu/smd/pathres/URPLM.html } "Most ass drivers are evil, most camel drivers are decent, most sailors are } saintly, the best among physicians is going to Gehenna, and the best of } butchers is a partner of Amalek" -R. Judah, in Mish. Kidd. 4:14 } } }
Could you please delete my subscription please. I apologise for not using the listserver address but due to circumstances beyond my control I lost the address.
Thanks for the replies on Electron flight simulator. Even a local vendor replied and I will be communicating with him. The demo version is a bit limited.
Thanks again ## [########] ## ## ## ## ## Stephan H Coeztee Electron Microscope Unit Private Bag 3 Wits 2050 South Africa
Dear Yves, I use a piece of glass, actually a cover slip, in place of the specimen to align my ion guns. The glass fluoresces under the ion beams and you can align the bottom gun and see it with the top gun off, then turn off the bottom gun and do the top. With the tantalum oxide, how will you see the bottom gun? The usual way to oxidize a metal is to heat it bright red (in a bunsen butner flame) then cool it in air. You wrote: } Dear all, } } Can anyone tell me how to oxidize surface of a tantalum disk in order to } perform the alignment of the guns in an ion mill. It must be something } simple but I do not know it. } } Thanks, } } Yves MANIETTE Regards, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
You will find attached an information for a postdoctoral position available in Laboratoire de pathologie Vegetale (Institut National Agronomique de Paris). Direct inquiries to Brigitte Vian or Yves Bertheau
Brigitte Vian INRA INA P-G, Pathologie Vegetale, 16 rue Claude Bernard, 75231 Paris cedex 05, FRANCE. Tel: +33 (1) 44.08.18.83 Fax: +33 (1) 44.08.16.31 E-mail: vian-at-inapv.inapg.inra.fr http://inapv.inapg.inra.fr/Welcome.html
Dear Yves, I use tantalum foil to align my Gatan guns. I oxidise the foil in 10% sulphuric acid, 90% water at 20Vdc at 20C in an electropolishing bath with the foil positive and a platinum strip negative. I stop when the foil is fairly dark purple (depends on the thickness of oxide you prefer). Regards Mike
Michael J Witcomb PhD Electron Microscope Unit University of the Witwatersrand Private Bag 3 WITS 2050 South Africa
Thank you to John McCaffrey, Jon Hangas, Mary Mager, Mike Witcomb, barbara foster, Brian G. Demczyk for their answers. Here is a synthesis.
The question was:
Can anyone tell me how to oxidize surface of a tantalum disk in order to perform the alignment of the guns in an ion mill? It must be something simple but I do not know it.
The answers are: **********************
A very effective way to align guns in an ion mill is to replace the tantalum disks with a thin glass coverslip. The glass fluoresces, showing the location of the ion beam(s). Make sure to make a mark on the glass where the center of the tantalum disks (i.e., your sample) would be.
You did not state what model ion mill you are aligning. Does tantalum oxide fluoresce? If not I would find it inconvenient to use for aligning ion mills.
It's been some time since I aligned a Commonwealth Scientific ion mill. I used to use zirconia, which fluoresced under the beam. I could adjust the screws holding the guns while watching the sample fluoresce, and try to get maximum brightness. On one home-built ion mill in graduate school, the sample stage was out of alignment and had to be shimmed.
If you are unable to obtain or make a zirconia foil, or other fluorescing material, try a heavy carbon coat on some material. It may be easier than oxidizing Ta.
A Gatan Dual Ion mill (which I use now) should not need alignment. However, you do not switch gun parts when you clean a Gatan Dual ion mill, otherwise the index marks on the epoxy end pieces of the gun assembly will not be properly aligned, and your cathodes will be off-center. Also, if the Whisperlock is designed as a cold stage, do not use it at room temperature without using the shorter room temperature platform. Otherwise your sample will be too high.
Regards, Jon Hangas jhangas-at-ford.com **********************
I use a piece of glass, actually a cover slip, in place of the specimen to align my ion guns. The glass fluoresces under the ion beams and you can align the bottom gun and see it with the top gun off, then turn off the bottom gun and do the top. With the tantalum oxide, how will you see the bottom gun? The usual way to oxidize a metal is to heat it bright red (in a bunsen butner flame) then cool it in air.
I use tantalum foil to align my Gatan guns. I oxidise the foil in 10% sulphuric acid, 90% water at 20Vdc at 20C in an electropolishing bath with the foil positive and a platinum strip negative. I stop when the foil is fairly dark purple (depends on the thickness of oxide you prefer).
E-mail: mikew-at-gecko.biol.wits.ac.za
[COMMENT from Ludo Rossou {ludo_gertie-at-ruca.ua.ac.be} : also works with 0.5g Na2SO4 in 300 cc water, increasing voltage up to 200 V DC. The green oxide layer will appear in a few minutes.] ********************** You must have an "Ion-Tech" mill! I've never found this procedure necessary in Gatan DuoMills, but dis have to go through this procedure (largely at the urging of the Ion Tech company rep.). They'll send you a pair of Ta plates for this purpose upon request.
"Brian G. Demczyk" {demczyk-at-erxindy.rl.plh.af.mil} **********************
Contact Bill Miller at Microbill-at-aol.com. He used to be product manager at both Zeiss for EM and President at Bal-Tec (ion mills). He knows lots of great tricks.
barbara foster {mme-at-map.com} **********************
I am interested in SEM JEOL 840 and have a question about scanning coil (cost, price etc.). because our scanning coil is damaged (burned and not work). however rest of SEM JEOL 840 is all right. Required scanning coil may be already used but still applicable (functional).
J o z e f Stankovic Faculty of Natural Sciences Comenius University Central Laboratory of Electron - Optical Methods Mlynska dolina 842 15 Bratislava Slovak Republic Europe
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am interested in SEM JEOL 840 and have a question about scanning coil (cost, price etc.), because our scanning coil is damaged (burned and not work), however rest of SEM JEOL 840 is all right. Required scanning coil may be already used but still applicable (functional).
J o z e f Stankovic
Faculty of Natural Sciences Comenius University Central Laboratory of Electron - Optical Methods Mlynska dolina 842 15 Bratislava Slovak Republic Europe
} Wearing a respirator is a lot better than nothing. But surely, pulverising } the tissue out on the open bench will cause fine particles to fly all over } the lab with every air current, contaminating everything and everybody in } the lab. I would use a biohazard hood.
I agree that using a hood will prevent particles from getting onto environmental surfaces. If you have a hood use it. On the other hand, I wouldn't be really concerned about infectious particles on environmental surfaces. The bugs I'm more concerned about spread by inhalation, not by skin contact. If you don't want to risk bringing something home with you, a surgical gown or similar smock can be used to protect your clothing.
Leon
-- Leon A. Metlay, M.D.,Associate Professor of Pathology and Laboratory Medicine University of Rochester Medical Center Phone: (716) 275-5691 P.O. Box 626 Fax: (716) 273-1027 Rochester, NY 14642 lmetlay-at-acu.pathology.rochester.edu http://www.urmc.rochester.edu/smd/pathres/URPLM.html "Most ass drivers are evil, most camel drivers are decent, most sailors are saintly, the best among physicians is going to Gehenna, and the best of butchers is a partner of Amalek" -R. Judah, in Mish. Kidd. 4:14
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Yves,
I use tantalum oxide disks to align gatan ion millers. I usually "punch" a large disk of tantalum foil, the size of the bottom plate/cover plate of the stage, and "punch" the three holes for mounting. I believe that a large sample is good for showing the profile of the ion beam as well as the location.
To oxidize, I make a salt solution (with any salt) and deionized water. Next, you need a dc power supply capable of rather high voltages, say 150 or 200 volts. I use a stainless steel rod to connect to the negative side of the power supply with an aligator clip. The tantalum disk goes to the positive side. Put the rod in the solution, set a voltage between 40 and 150, and dip the disk in. Be careful not to touch the disk and the rod. Also, avoid getting the aligator clip in the solution. The oxide layer thickness will saturate depending on the voltage and will reach a corrosponding color. Very dramatic colors can be acheived. We have actually made a chart of different voltages and their representative colors. As the oxide is removed it leaves behind the characteristic color you can look up on the chart. You can now get a measure of the milling rate with very short milling times, 10's of seconds. You can re-electroplate a "used" disk to get the original color or increase the voltage a little and re-plate the entire disk.
The colors you get depend also on the initial surface condition and thickness of the tantalum. We use 250 micron thickness.
Have fun, mike.
Michael T. Marshall Research Engineer, Electron Microscopy University of Illinois at Urbana-Champaign Frederick Seitz Materials Research Laboratory 104 South Goodwin avenue Urbana, IL 61801-2985 (217) 244-8193 fax: (217) 244-2278
There has been much discussion on this topic in the last year or so and I have archived it all at the web address at the end of this message. Go to the "Tips & Tricks" link and in there find the "Photography" section. I believe there are a couple of links you will find interesting.
In answer to your question we have had an Ilford 2150 which has been up and down since we have owned it with little minor repairs. As far as daily maintance goes, there is little. Simply turn it on and go. Chemicals need relacement every 3 weeks or 1000 8x10 prints, whichever comes first. It just seems to need gears, bearings and bushings replaced every so often. Cheap but a hassle. Hope this helps.
At 03:15 PM 2/17/97 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 352-392-1295 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
You indicated that you would be grinding up the tissues. Why not embed in paraffin and then cut slices which could be attached to an inert substrate, deparaffinized and examined in the SEM. This would keep aerosols to a minimum - if not eliminate them entirely by encapsulation in paraffin during the critical cutting stage.
} } } Our lab has been asked to characterize lyophilized human lung tissue by } } } SEM and XRF analysis. Not being pathologists and having no prior } } } experience with biological tissue samples, we are concerned about } } } potential health risks from handling this material. The samples were } } } collected and lyophilized at least 20 years ago and have been in storage } } } all this time.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 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've received a number of questions/comments regarding my postings about my LINK conversion software. Because of time constraints, and on the asssumption that the answers may be of interest to others too, I'm replying to everyone via this posting. Apologies to other subscribers, who are clear about or uninterested in the information. This is fairly long, so hit "Delete" now if you are not interested.
Just to be clear, I am a University user with no connection, other than as a customer, to any equipment manufacturer. The comments below about Link's products were gleaned from documentation, experience and conversations with Link's employees. Obviously the comments offered are correct to the best of my knowledge, but they are offered on an "as is" basis, for what use you care to make of them.
Before the ISIS system, Link analyzers were based on proprietary computers which owed a lot to the Data General Nova systems (the Link 290 was actually built around a Nova, I believe). It is my understanding that the backplane pin configuration was the same as the Nova (but the timing may have been different), and the disk directory and file format was the same as the Nova. The compatability may, for all I know, have been compromised as Link developed their products, but I was able to write very simple machine code programs using my old Nova 3 and it's assembler, which would execute on the AN10000.
I know very little about Link products before the AN10000. The early AN10000's had a disk operating system called "Demon". Later this was upgraded to become "Demon-Plus" which would only execute on hard-disk versions of the AN10000 (our first AN10000 had two 8" floppy drives - it still has one!). The operating system of the eX/L was called "Genie".
All of these wrote files onto disk using the Data General format, which Link thoughtfully described in detail in the AN10000 system manager's guide. Link have always also described the details of their proprietary file formats (i.e. which byte stores what information) in their software manuals.
From the early days of the AN10000 customers have been frustrated by the difficulty of exporting data from their Link analysers, and the company developed an range of products designed to transfer data to other computers. These were not particularly sophisticated for the AN10000, but on the later eX/L's they do a fine job. I have used Mac-Link, for example, and it transfers images, linescans, etc. to a Mac while preserving colour information. The later AN10000's and eX/L's also had a program which would copy Link's files to a MS-DOS 3.5" floppy disk, but this works very slowly (I think because it has extensive error checking built in).
Having got on to colour, I can only speak for the AN10000 (I have an eX/L but haven't looked into it in such detail). A linescan is stored as a one-dimensional image - i.e. a sequence of numbers representing, for example, the number of x-ray counts or the intensity of the electron signal. When you run the image analysis program (Digipad, in the case of the AN1000) the linescans are plotted on the screen according to the colour capabilities. Our AN10000 can only display 16 colours, so Digipad converts everything to 16 colours. It uses an indexing and lookup table scheme to generate the image. If you export the image from Digipad and look at it with another program using a different lookup table, you will get different colours. For example, colour coded linescans, when exported and viewed, let us say, with Photoshop with a 256-bit grey-scale lookup table will appear to have the plots in differing shades of (very dark) grey.
What my software allows is:
1: exporting spectra so thay can be read by any software that understands them
2: conversion of the formats of the Link files so that images are stored as TIFF files and spectra and linescans can be stored as ASCII files. They can then be read into software such as image presentation or analysis packages or spreadsheets, running on fast modern computers, so I don't tie up the microscope while I am processing my data.
3: extraction of the individual images from studies (I don't know what would happen if you tried to extract a single linescan from a linescan study - I've not tried it).
If I want colour then I do all the processing on the PC. I don't try to export Link's files. I use Photoshop or a similar programme for images, and a spreadsheet or graphing programme for linescans. I import my spectra to DTSA on the MAC, but there are other packages too that will import Link binary spectrum files. Alternatively, if I want a well-plotted spectrum with no analysis, I use the ascii formatted output from LKCV and read it into a graphing package.
Everything I have is on the FTP server, including all the source, comments, etc. Time constraints prevent me from offering any more support, although I don't mind answering simple questions by e-mail (I'd prefer to do this on the server, as then I won't get asked the same thing 20 times over!). As you can see from the file dates, some of the code is several years old.
Hope this all helps!
Enjoy!
Tony Garratt-Reed
**************************************************** **************************************************** ** ** ** Anthony J. Garratt-Reed ** ** Room 13-1027 ** ** Center for Materials Science and Engineering ** ** Massachusetts Institute of Technology ** ** 77 Massachusetts Avenue ** ** Cambridge, Massachsetts 02139-4307 ** ** U. S. A. ** ** ** ** Phone: 617-253-4622 ** ** Fax: 617-258-5286 or 617-258-6478 ** ** ** **************************************************** ****************************************************
Dear Jonathan: It may be that the filter insert in your existing oil mist filter has reached saturation. The need to change the insert will depend your frequency of use. The amount of oil vapor passing through a good quality clean filter should be minimal and the health problem probably not serious if entering a room with adequate air turnover. I agree that if you can smell the oil it's undesirable. Depending on how much you run the evaporator, I would be as concerned about my mental state. The noise from those things can be nerve-racking!
Don Gantz Boston Univ Med School gantz-at-med-biophd.bu.edu
I want to ask if anyone works paraffin shrimp processing and microtomy. I have troubles in shrimp paraffin sectioning, because of the hard skeleton of the shrimp. How could I soften the exoskeleton of the body of the shrimp ?. How could I soften the chitinous exoskeleton of the shrimp to make paraffin sections ?.
The glass target suggested by Mary Mager sounds like the superior method. Here are a few reasons for thinking so:
1. It allows aligning both guns without removing the target.
2. One glass target can be used repeatedly without wearing out. The anodized Ta foil is essentially a one-shot method because it works by wearing the oxide surface layer off.
3. The glass target allows the ion guns to be adjusted dynamically, whereas the Ta foil is used quasi-statically. i.e., you expose the Ta foil to the ion beam, raise it to the viewing position (in the Duo-Mill), adjust the gun, sometimes have to replace the target, and reiterate until satisfied (or frustrated). The erosion pattern can be hard to see through the viewport, and the foil targets sometimes need to be replaced one or more times during a complete gun alignment.
4. It requires no special equipment, no chemical handling facilities, and no HV power source to make.
Ion mill manufacturers I know of abandoned the anodized foil target in favor of a scintillator target long ago.
Larry Thomas Mechanical and Materials Engineering Washington State Univ. thomas-at-mme.wsu.edu ------------------------------------------------------------------------------ REPLY FROM: Thomas, Larry Return-Path: {Microscopy-request-at-Sparc5.Microscopy.Com} Message-Id: {199702181528.JAA01758-at-Sparc5.Microscopy.Com} X-Sender: marshall-at-uimrl3.mrl.uiuc.edu Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Yves,
I use tantalum oxide disks to align gatan ion millers. I usually "punch" a large disk of tantalum foil, the size of the bottom plate/cover plate of the stage, and "punch" the three holes for mounting. I believe that a large sample is good for showing the profile of the ion beam as well as the location.
To oxidize, I make a salt solution (with any salt) and deionized water. Next, you need a dc power supply capable of rather high voltages, say 150 or 200 volts. I use a stainless steel rod to connect to the negative side of the power supply with an aligator clip. The tantalum disk goes to the positive side. Put the rod in the solution, set a voltage between 40 and 150, and dip the disk in. Be careful not to touch the disk and the rod. Also, avoid getting the aligator clip in the solution. The oxide layer thickness will saturate depending on the voltage and will reach a corrosponding color. Very dramatic colors can be acheived. We have actually made a chart of different voltages and their representative colors. As the oxide is removed it leaves behind the characteristic color you can look up on the chart. You can now get a measure of the milling rate with very short milling times, 10's of seconds. You can re-electroplate a "used" disk to get the original color or increase the voltage a little and re-plate the entire disk.
The colors you get depend also on the initial surface condition and thickness of the tantalum. We use 250 micron thickness.
Have fun, mike.
Michael T. Marshall Research Engineer, Electron Microscopy University of Illinois at Urbana-Champaign Frederick Seitz Materials Research Laboratory 104 South Goodwin avenue Urbana, IL 61801-2985 (217) 244-8193 fax: (217) 244-2278
we used to order, for our customers who have TEM with low level camera, the Dage MTI DSP 200 enhancer video signal. This product is now obsolet. Does someone know the same kind of product to increase brightness and contrast and make real time averaging on video signals ?
I concur with the others that using a microscope cover slip is an excellent means of aligning ion sources. Aside from it being easily cut with an ultrasonic disk cutter and fluorescing rather nicely, its thickness is similar to that of a TEM specimen. The cover slips that we use are about 75 microns in thickness. It is important to use as thin a slip as possible. Because you are aligning the beams based on the fluorescence from the surfaces of the cover slip, the relative beam position to the center of the specimen will change slightly as the specimen approaches perforation. For the most part this shift is negligible, however, should real thick (} 200-300 micron) pieces of glass be used, the alignment could be adversely effected.
Regards,
Paul Fischione
E.A. Fischione Instruments, Inc. produces both an ultrasonic disk cutter and ion mill used for TEM specimen preparation.
Can anyone tell me how to oxidize surface of a tantalum disk in order to perform the alignment of the guns in an ion mill? It must be something simple but I do not know it.
I was wondering if there are other JEOL 100CX users out there who also have a problem with their shutter system. 70% of my service calls involve the stage not comming up to 90 degrees, hence cutting the bottom third of my negatives. I have had the stage motor replaced on two occasions and just yesterday two of the screws holding the motor in were out. According to the JEOL service manager this is a bad design....which is wonderful to find out after you have taken fifty useless negatives. Please post me privately if you are uncomfortable with a general broadcast.
John Grazul Rutgers University Electron Microscope Facility
National Institutes of Health Bethesda, Maryland Biomedical Engineering & Instrumentation Program Analytical Electron Microscopy Laboratory
Physical /life scientist (Biologist GS9-GS11) at BS or MS level with solid experience in thin-section transmission electron microscopy. Experience in advanced techniques in analytical electron microscopy and structural biology (cryosectioning, freeze fracture, and macromolecule preparation) is desirable, although on-the-job training is possible for an experienced and meticulous microscopist. PROOF OF U.S. CITIZENSHIP IS REQUIRED for this appointment.
For further information please contact:
Dr. Richard Leapman Bldg. 13, Rm. 3N17 National Institutes of Health Bethesda, MD 20892 Tel: (301) 496-2599 FAX: (301) 496-6608 E-mail: leapman-at-helix.nih.gov
Applications (with curriculum vitae or federal employment form SF-171) should include the reference number RR-97-0008A, and should be post-marked by 3/10/97 and sent to: Mr. Eugene McDougal Bldg. 31, Rm. 3B38 National Institutes of Health Bethesda, MD 20892-2130 Tel: (301) 496-1524 (for detailed instructions about application)
You can retrieve more information via FAX by calling 301-594-2953 (local) or 1-800-728-JOBS (long distance) -- FAX-ID# 5808. General information is also available at http://www.nih.gov/news/jobs/
In re-reading my post regarding my screen problems on my JEOL 100CX I made a erroneous statement {I did not edit my posting before it was sent}. I think that this is a bad design because of the cronic problems that we have had over the eight years that we have owned the scope...unfortunately I added a false statement regarding our service manager. This is a result of a quick send finger and faulty editing on my part. I apologize to JEOL for my misstatement, but the stage drive still acts up cronically
John Grazul Rutgers University Electron Microscope Facility
The School of Materials Science and Engineering at Georgia Tech is seeking to obtain a double tilt holder for a Phillips 400 TEM. If you have such a holder for sale or trade or donation in your lab, please contact me by e-mail or phone. Regards,
Yolande Berta School of Materials Science and Engineering Georgia Institute of Technology 778 Atlantic Drive Atlanta, GA 30332-0245 (404) 894-2545 (404) 894-9140 FAX yolande.berta-at-mse.gatech.edu
Greetings! I have a Reichert-Jung Ultracut that still functions well after 20 years. However, it seems that one of the prisms in the American Optical Model 570 binocular head has become dislodged, making simultaneous focus impossible. It is not so noticable at low mag, but at high mag becomes very apparent. The person who services the instrument is reluctant to take the head apart. I'd like to know if someone could recommend a company or individual that could make the necessary repairs, without having to trade my son for the service. Thanks in advance!
Dwight Beebe E-mail: beebed-at-ere.umontreal.ca Institut de recherche en biologie vegetale Voice: 514-872-4563 Universite de Montreal FAX: 514-872-9406 4101, rue Sherbrooke est Montreal, Quebec H1X 2B2 Canada
OOPS! The server cut off the contact phone number and e-mail address, for that double tilt holder which you may have sitting around in your lab, for a Phillips 400. Let's try again. Yolande Berta School of Materials Science and Engineering Georgia Institute of Technology 778 Atlantic Drive Atlanta, GA 30332-0245 (404) 894-2545 (404) 894-9140 FAX yolande.berta-at-mse.gatech.edu
(1) I am interested in any references on scanning EM of polyacrylamide gels of any type. My main interest is in the structure of these gels. I would like to know what the "pores" look like. A medline search from 1966-present was negative.
(2) If anyone out there has looked at polyacrylamide gels of any type, I would be interested in unpublished results, if you are willing to disclose your findings.
(3) Finally, if anyone is interested in a colloboration to look at these, please contact me at the address below. I am currently making my own tube gels, but I also use commercially prepared 10-20% tricine gels. Concerning the "home made" gels, I am particularly interested in effects of acrylamide concentration, crosslinker concentration and temperature on the structure of the gel.
Thanks! ------------------------------------------------------------------------ Sheryl K. Brining, Ph.D. Bldg. 10/Room 6C-103 National Institutes of Health Bethesda MD 20892-1582 National Institute on Aging e-mail: skb-at-helix.nih.gov 10 Center Drive, MSC 1582 Phone/Fax: (301) 594-3982
"Whatever you can do, or dream you can, begin it. Boldness has genius, power and magic in it." Goethe
We use DVS-3000 by HAMAMATSU photonics. This product features integration, enhancing. You may need to make sure that the averaging function is as strong as you wish. They also have a new model DVS-20 which is controlled with SCSI.
Takanori Maeda Pioneer Electronic Corp.
} Hi all, } } we used to order, for our customers who have TEM with low level camera, the } Dage MTI DSP 200 enhancer video signal. This product is now obsolet. Does } someone know the same kind of product to increase brightness and contrast } and make real time averaging on video signals ? } } Elisabeth LECA } NEWTEC/Assistance } }
} Can anyone point me to a software vendor from whom I can buy a copy } of the ImagePro image processing program?
The homepage of Mediacybernetics should help You: http://www.mediacy.com/ -- Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 93 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www_scem.csb.ki.se/pages/philip.html
Calling all manufacturers/major distributors of microscopes/related products.
Indian economy has recieved a boost by libralisation of imports and globalisation today.Protective tade measures and other hurdles have been removed making imports easier.
There is a good ready market for the following areas of microscopy.
1.Light microscopy. 2.Image analysis 3.EM/SEM 4.aCCESSORIES LIKE Optics-objectives,eyepieces,condensors,phase contrast eqpt.,polarising equipt.attachments. 5.CCD Cameras,video printers,Image grabber cards 6} Training courses
Organisations associated in the above areas interested in exloiting the Indian market by way of appointment of distributors/representation/ technical collaborations for manufacturing/outsourcing/sub contracting may kindly send their product catalogues,views on this possibility immediately to the undersigned.
An early response shall be highly appreciated.
Best regards, Soneja A.K. ************************************************************************* For further details please contact: Soneja A.K. Director METZER BIOMEDICAL & ELECTRONICS PVT.LTD. 327 Wadala Udyog Bhavan,Wadala,MUMBAI(BOMBAY )400 031.INDIA Tel:91 22 4145057/4165650 Fax 91 22 4168757
What is the advantage of a cold FEG vs. a Thermal FEG for a 200 keV TEM? Is the stability of a cold FEG a real issue? Does the thermal FEG have an edge for operation in TEM imaging? Is the Cold FEG superior for small probe work? Are there any other compelling issues in deciding about a Hitachi vs. a Jeol/Philips TEM?
These are the only papers I know of that show EM of polyacrylamide:
R=FCchel, R. and M.D. Brager. 1975. Scanning electron microscopic observations of polyacrylamide gels. Anal. Biochem. 68:415-428.
R=FCchel, R., R.L. Steere, and E.F. Erbe. 1978. Transmission-elect= ron microscopic observations of freeze-etched polyacrylamide gels. J. Chromatogr. 166:563-575.
We are interested in the exclusion of macromolecules from gels, and I= would be interested in your findings on the ultrastructure of your gels.
Jim Williams.
/////////////////////////////////////////////////////////////////////= ////// / James C. Williams, Jr. williams-at-anatomy.iupui.e= du / / Department of Anatomy = / / Indiana University School of Medicine (317)274-3423 = / / 635 Barnhill Drive (317)278-2040 fax = / / Indianapolis, IN 46202-5120 = / /////////////////////////////////////////////////////////////////////= ////// Great are the works of the LORD, studied by all who have pleasure in them. Psalm 111:2
Sheryl K. Brining wrote:
} (1) I am interested in any references on scanning EM of polyacrylam= ide } gels of any type. My main interest is in the structure of these gels= . I } would like to know what the "pores" look like. A medline search fro= m } 1966-present was negative. } {snip}
Has anyone used Green Fluorescent Protein? I would like to use it for = determining cell proliferation (because of the experimental set up, it = is difficult to measure cell population directly).
Also, does anyone know of any dyes that do not affect cell viability but = also will be retained by the cell long term (~2-3 weeks) without being = metabolized or compartmentalized?
Eric, Molecular Probes has a great web site - http://www.probes.com that you may find helpful or call them for technical help at 541-465-8353 if you don't have access to the www.
Beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
The Electron Phase Problem: 1) Obtain an Image/Diffraction Pattern, probably of the sample 2) Obtain a probable electron wave 3) Obtain a probable structure 4) Probably the referee will accept it
Abstract Due by March 15 See http://www.msa.microscopy.com/MSAMeetings/MM97link.htm
Preliminary list of invited speakers: J. E. Bonevich "Electron Holography of Electromagnetic Fields" C. Barry Carter "Fresnel-Fringe Contrast from Interfaces" D. L. Dorset "The pseudo-atom approximation in direct determination of protein structures" C. Gilmore "The Maximum Entropy Method for Solving the Phase Problem" B. K. Jap "3D structure of a water channel at 6.5 A resolution as determined by electron crystallography" S. Hovmoeller and X. Zou: "The relation between the phase of the electron wave (which is lost when an EM image is recorded) and the crystallographic structure factor phase (which is present in the EM image)" C. A. Mannella "3D structure of a mitochondrial ion channel by electron crystallography" L. D. Marks "Is it really that easy to solve Surface Structures using Direct Methods?" I. Voigt-Martin "The use of electron crystallography to solve old problems in non-linear optics" N. Tanaka "Possibility of coherent CBED of bi-crystals and its multi-slice simulation" H. W. Zandbergen "The use of the phase and and amplitude of exit waves for accurate structure determination"
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University Evanston, IL 60208-3108 tel: (847) 491-3996 fax: (847) 491-7820 email: l-marks-at-nwu.edu http: //www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
} Date: Wed, 19 Feb 1997 14:51:21 -0400 } From: Dwight Beebe {beebed-at-ere.umontreal.ca} } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Service for Ultracut binoculars } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Greetings! } I have a Reichert-Jung Ultracut that still functions well after 20 } years. However, it seems that one of the prisms in the American Optical } Model 570 binocular head has become dislodged, making simultaneous focus } impossible. It is not so noticable at low mag, but at high mag becomes } very apparent. The person who services the instrument is reluctant to take } the head apart. I'd like to know if someone could recommend a company or } individual that could make the necessary repairs, without having to trade } my son for the service. } Thanks in advance! } } } Dwight Beebe E-mail: beebed-at-ere.umontreal.ca } Institut de recherche en biologie vegetale Voice: 514-872-4563 } Universite de Montreal FAX: 514-872-9406 } 4101, rue Sherbrooke est } Montreal, Quebec H1X 2B2 } Canada } I have had good service on both the microtome and the binocs from Jon Petz (VP) at
I am looking for TEM related job. If you can provide imformation about this or show me the way to get the imformation, I will preciate it. Thank you very much.
Organized on behalf of the Royal Microscopical Society by: Prof Anthony G Cullis (a.g.cullis-at-sheffield.ac.uk) Dr John L Hutchison (john.hutchison-at-materials.ox.ac.uk)
Co-sponsored by the Institute of Physics (EMAG) and Endorsed by the Materials Research Society
CENTENARY OF ELECTRON DISCOVERY ------------------------------------------------------------
The conference will feature a special Symposium to celebrate the centenary of the discovery of the electron. Key presentations will be given by: Dr W F Brinkman (Vice-President for Physical Sciences Research, Bell Labs, Murray Hill) The Materials Behind the Telecommunications Revolution Dr T Matsuo (Managing Director of Semiconductor Equipment Division, JEOL Ltd, Tokyo) The Evolution of Semiconductor E-Beam Lithography and Metrology Prof K van der Mast(Philips Electron Optics, Eindhoven) The Development ofElectron-Optical Imaging and Diffraction Systems
MAIN CONFERENCE SCIENTIFIC SESSIONS
These will focus on the state-of-the-art in studies of the structural, electronic and optical properties of as-grown and processed semiconductors by all forms of microscopy. More than 160 papers will be presented and the full scientific programme is available at the RMS Web Site http://www.rms.org.uk. The proceedings of the conference will be published.
INVITED SPEAKERS -----------------------------
Prof P J Goodhew (University of Liverpool) Dislocation Behaviour in Strained Layer interfaces Prof R J Hamers (University of Wisconsin-Madison) STM Studies of CVD Processes on Si Surfaces Dr D C Houghton (Canadian National Research Centre, Ottawa) Advances in Epitaxial Strained Layer Devices Dr D E Jesson (Oak Ridge National Laboratory, Tennessee) Exploring Instabilities and Metastabilities in Semiconductor Growth Dr J-L Rouvi=E8re (CEN, Grenoble) GaN Growth: Influence of Polarity and Strain Prof J C H Spence (Arizona State University) Dislocation Kink Behaviour in Semiconductors Prof H P Strunk (University of Erlangen) Self-Organization and Defect Mechanisms in Heteroepitaxial Growth Prof S Takeda (University of Osaka) The Structures of Extended Defects in Si and Ge Analysed by HRTEM Dr R T Tung (Bell Laboratories, Murray Hill) Control of Silicide Layers in ULSI Devices: Simple Principles at Work Dr J Vanhellemont (IMEC, Leuven) TEM Studies of Processed Si Device Materials Dr P R Wilshaw (University of Oxford) Developments in SEM:EBIC Studies of Semiconducting Materials
********************************
Further details, including registration information, can be obtained from: The Administrator, The Royal Microscopical Society, 37/38 St Clements, Oxford OX4 1AJ, UK Tel: +44-(0)1865-248768 Fax:+44-(0)1865-791237 E-mail: meetings-at-rms.org.uk WWW: http://www.rms.org.uk
GFP is good, but it knocks out most of the green dyes, if you want to double label anything, since it has a broad spectra. PKH from sigma is another choice for tagging, etc., but I think that goes only to 10 days. Good luck.
Someone just asked about the nature of the Pentavac-5 DP oil that has just been listed as a new item in the catalog of the Duniway Stockroom Corp. Here is a comparison of the properties published for it by Duniway with those published by Monsanto for Santovac-5: Pentavac Santovac Vapor Press -at- 25 C 4x10-10 Torr 4x10-10 Torr BP at 0.5 Torr 275 C approx. 260 C Viscosity -at- 40 C 279 -at- 38 C 360 Viscosity -at- 100 C 12.6 -at- 99 C 13
Although all values are not identical, you must remember that it is very difficult to measure the properties of fluids such as these with a high degree of precision, and so my suspicion is that Pentavac-5 is essentially the equivalent of Santovac-5, but probably produced by a company other than Monsanto. My understanding is that Monsanto recently announced that they intended to stop manufacturing Santovac-5, and that they were looking for another company to take over the operation. My guess is that this transfer has now occurred, and that Pentavac-5 is the result. Incidentally, when first introduced in the early 1960s the prices quoted for Santovac-5 by Monsanto were: $11.50 for a 100 ml bottle and $43.00 for a 500 ml bottle. Current prices for Santovac-5 are $150 and $560, and for Pentavac-5, $130 and $510 - how times have changed!
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
We are planning a study of airway permeability in response to tobacco smoke in the lung; that is, fluid transport from blood vessel through endothelium to interstitium through epithelium to lumenal airspace. We would like to use a marker that we will be able to identify with the high-resolution low-temperature SEM in frozen hydrated fractured specimens of hydrated airways. The pore size for this pathway is greater than 50 nanometers diameter.
Would colloidal gold work? It is available in an appropriate range of sizes but is it sticky, will it attach along the way? How about ferritin? Can it be identified at reasonable magnification in frozen hydrated specimens with backscattered electrons? Does it agglutinate into too large complexes?
Any suggestions from you will be appreciated.
Jacob
Jacob Bastacky, M.D. Room 116 Donner Laboratory Lawrence Berkeley Laboratory EMail: sjbastacky-at-lbl.gov University of California Phone: (510) 486-4606 Berkeley, California 94720 Fax: (510) 486-4750
Thanks to all for the quick responses about my search for Image Pro! From the many notes I received, I should be able to get all the information I need.
Rob Plano Staff Analyst, SPM Services Charles Evans & Associates Sunnyvale, CA (408) 739-3867, ext.294 rplano-at-cea.com
Hello all. I know I've seen this topic come up here before, but now that I need it, I can't find the information.
Can anyone recommend some solvents, other than alcohols and acetone, suitable for freeze-substitution. I am having some difficulty obtaining adequate substitution in some cells of my material (plant bits) and want to try solvents which may penetrate more effectively.
Tetrahydrofuran is a great one to try. it gives very different results than acetone or ethanol. But I doubt penetration is the problem. Why do you think you are getting bad penetration?Are you sure your solvents are totally dry? I recommend using a fresh unopened bottle each time. Avoid adding molecular sieves which can make matters worse.
} } Can anyone recommend some solvents, other than alcohols and acetone, } suitable for freeze-substitution. I am having some difficulty obtaining } adequate substitution in some cells of my material (plant bits) and want to } try solvents which may penetrate more effectively. } } Thanks in advance. } } Kim Rensing
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
} I am looking for TEM related job. If you can provide imformation about } this or show me the way to get the imformation, I will preciate it. Thank } you very much.
A good site is Microworld Resources http://www.mwrn.com/ The of course the job adds in Science, Nature etc. and the postings on this list. -- Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 93 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www_scem.csb.ki.se/pages/philip.html
Scanalytics, the biological imaging division of CSPI, is looking to add to our Technical Support team. The position is for our microscopy product line which includes software-based instruments for performing high-resolution 3D fluorescence microscopy.
The position requires that you interact with potential and current customers to answer their questions on how the Scanalytics products may be configured to be most appropriate for their work and to perform benchmark studies. You will be called upon to perform demonstrations, and installations of the instruments, to deliver technical presentations to interested audiences, and to travel with sales representatives (domestic and international) to answer technical questions. Your experience will be needed to help in the design of new products and to write technical bulletins regarding current products. In addition to a competitive base salary, health/life insurance, and tuition reimbursement, compensation includes a commission on product line sales.
While this position does not require direct experience in the field of microscopy, candidates will possess strong interpersonal abilities, a desire to learn new skills in a dynamic industry, a willingness to travel, and a problem-solving attitude. Ideal candidates will bring several years of hands-on experience with biological fluorescence microscopy, especially computer-based (PC) instruments as applied in this work, a good working knowledge of research cameras and microscope automation equipment is also desired. The position will involve approximately 50% travel away from our Boston-area office. A BS in the life sciences or biomedical engineering is also highly desirable.
Persons interested in exploring this exciting opportunity are encouraged to send their resume to:
Rose Doyon Scanalytics 40 Linnell Circle Billerica, MA 01821
Larry Stoter posted a list of parameter for thermal vs cold FE emitters, including the following:
Energy Spread ev 0.2 - 0.4 0.3
These numbers are commonly seen in the literature, but they are only good for very low beam currents, say at 1 microamp emission, which is not useful for "typical" microscopy. At more realistic currents encountered e.g. for high resolution imaging (say 30 microA), the cold FE source will operate at 0.5-0.6 ev (200kV), and the Schottkey source significantly higher. Don't believe everything you read.
Several folks have asked for the response to my query about staining silicone residues in tissue. Thanks so much for all the help and please excuse my tardiness in getting this out to the members.
Regards, Don Cox
- - - - - - -RESPONSES- - - - -
Someone, a week or so ago (I deleted the message), asked for info about staining of silicone in tissue. I happened to come across the following reference which may be of interest: Raso D et al. 1994. Light microscopy techniques for the demonstration of silicone gel. Arch. Pathol. Lab. Med. 118: 984-987. There also was an accompanying editorial in the same issue by Roggli et al. - Daniel Luchtel {dluchtel-at-u.washington.edu}
- - - - -
Reply to: RE} Staining Silicone-Containing Tiss We have been staining breast tissues from silicone implants for the past 5 years. We receive frozen tissue and cut slides for an immunofluorescent panel. We do IgA, IgM, IgG, C3 and Fibrinogen. The sections are cut at 4 microns and there have not been any cutting problems. Carol Ann Bobrowitz {Carol_Bobrowitz.PATHOLOGY-at-qmail.path.mcw.edu} Medical College of Wisconsin
- - - - -
Dear Don and Histonetters: There is an excellent reference entitled " Light Microscopy Techniques for the Demonstration of Silicone Gel" in the October, 1994 Volume 118 Issue of The Archives of Pathology and Laboratory Medicine. I had the opportunity to hear the author, Dr. Dominic Raso, speak on this subject. Bottom line -- Non-koehler, phase contrast and darkfield illumination greatly enhance detection of silicone gel. He also mentions a negative staining technique which is a 1 : 1 mixture of Aqua-mount and black stamp pad ink. Oil red O is not very consistent. Electron probe microanalysis also confirms the presence of silicon. Also, sections need to be cut at 10 micron. My own experience with silicone gel has been that it is highly refractile, not polarizable and no special stain was needed. If you have difficulties locating the reference, I will be glad to send you a copy. Linda Jenkins {jlinda-at-ces.clemson.edu} MUSC
- - - - -
I once had to look for Si in the breast tissue of a woman who thought she had been "poisoned" by her implants. I didn't find anything in the breast tissue itself.
Almost forgot..... I did this with EDAX in the SEM, looking at a section cut onto a plastic (Thermanox) coverslip, dried and carbon coated. My explanation would be a bit fuller but I'm in the middle of a conference. As I remember, a few relevant papers came up when I did a lit. search using keywords like breast, silicon, etc.
Get back to me if you want more, I'll be freer next week.
Stephen Edgar Electron Microscope Unit, Pathology Department School of Medicine University of Auckland
some time ago I posted a query about not being able to run Iomega ZIP parallel port drive under Novell DOS, only under MS-DOS We now found a solution which is perhaps useful to those who were interested: - instead of the recommended installation from floppy diskette using setup.exe, use following procedure: - boot machine from floppy drive into MS-DOS ( this must be available for the first installation); install ZIP drive, and run guest program. - once machine recognizes the ZIP drive, run install.exe from dosstuff on tools disk even though the drive is parallel port, the program will install some SCSI drivers; - machine can now be booted normally into Novell DOS, and will recognize ZIP Prof.Walter A.Mannheimer Metallurgy and Materials Engineering Federal University of Rio de Janeiro POBox 68505 21945 Rio de Janeiro, Brazil Voice +5521 280-7443 (Dept.office) +5521 590-0579 (direct) Fax +5521 290-6626 Email: wamann-at-metalmat.ufrj.br
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Here is a table of cold v Schottky FEG parameters
Cold Schottky Vacuum {10^-10 {10^-8 Tip flashing? 6-8 hr no Beam Noise 6-10% 1% Max Brightness 10^9 10^8 (A/cm2/sr) Current 1pA-300pA 1pA-5nA Lifetime } 1000 hr } 2000 hr I Stability/hr % {5 {1 Energy Spread ev 0.2 - 0.4 0.3 Source dia 5 - 10 nm 15 nm Work function 4.5 eV 2.8 eV Operating temp 300 K 1800 K
Data from "The Principles & Practise of Electron Microscopy" by Ian Watt.
I am looking for studies performed on reflection contrast microscopy (sometimes also named: interference reflection contrast microscopy). More than 200 citations can be found in literature but I am uncertain whether this technique is used somewhere routinely. Is anybody doing research work (or has done) with this equipment? And - if so - I would appreciate information by which manufacturer it has been produced.
-- Mit freundlichen Gruessen Yours sincerely ***************************************************************** * Dr. T. J. Filler * specialist in anatomy * * Westfaelische Wilhelms-Univ. * phone:*49 251 83 55226 * * Institute of Anatomy * FAX.: *49 251 83 55241 * * Image Analysis Division * e-Mail: filler-at- * * Vesaliusweg 2-4 * e-Mail: image.analysis-at- * * D-48149 Muenster * e-Mail: Institute.of.Anatomy-at- * * Germany * domain: uni-muenster.de * * http://medweb.uni-muenster.de/institute/anat/ * *****************************************************************
Loren Prentise wrote: ============================ I have a thin (1um) al2o3/al film on an aluminum (1mm) substrate. I am trying to mount for the cross section. I need a method to preserve the edge and prevent film spallation and substrate smearing while polishing. I have tried to sandwich film with another Al piece, but can't get intimate contact for good support, so film is badly damaged. One suggestion was electroless plating or electroplating. Any suggestions or resources to pursue? Thanks very much. ============================= If the object is to look by SEM, then it is correct that if diamond knife thin sectioned, throwing away the sections, but saving the "faced-off-piece" for examination, that surface from the "faced-off-piece" is going to be better than any surface that could be obtained by "polishing". The sample for this kind of work is generally embedded, and if the pore size is especially large, then we use vacuum embedding methods for even better results.
If you think (as we do) that it is a shame to be throwing out perfectly good sections, take a look at them by TEM. However, in that case, we always sputter coat a layer of gold on top of the anodized layer first. It does not interfere with the SEM examination of the faced-off-piece but it does help in the interpretation of the TEM results. If your interest is primariy in the top most surface of the anodized layer, then by all means embed. There might be some separation at the oxide/substrate interface, but there is excellent preservation at the interface. If a pice of the oxide is missing, you can readily tell from the presence of the gold layer whether that is real (e.g. oxide really was missing) or an artifact (it was pulled out during the sectioning). If the interest is more over all relative to the anodized layer, then sometimes it is better not to embed. Just gold coat and section. The presence or absence of the gold layer again helps to discriminate between fact from artifact, that is, whether a "hole" in the anodized layer is "real" or one caused by the diamond knife during sectioning.
Disclaimer: SPI Supplies offers materials science diamond knives and therefore we have a vested interest in seeing this kind of work done by diamond knife thin sectioning than by any other way!
Chuck
==================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ =====================================================
We have used an Agfa DD3700 processor for many years with excellent results; almost no down-time.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (908)-235-4583; fax -4029 e-mail: mcauliff-at-.umdnj.edu ***************************************************************
I also have had very good service from Tek-Net in New Jersey; 908-905-5530.
Geoff *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (908)-235-4583; fax -4029 e-mail: mcauliff-at-.umdnj.edu ***************************************************************
Regarding rotary pump oil vapors in areas where one cannot vent the pumps outside. I had posted a reply to J. Krupp about a double filter set-up I had constructed while serving time at U of Ill. This design has worked well for such an application.
Several others had inquired as to the construction of such a unit. I have a drawing available for the do-it-yourselfer or would be willing to construct one (or several) and send it out for a modest price to anyone interested. Please contact me via email: edb-at-chem.psu.edu if you would like further information on such a unit. If there is enough interest I may just quit my day job and produce these full time.
cheers Ed Basgall, PhD Penn State Univ Dept of Chem State College, PA 16802 Ph: 814-865-0493 FAX 814-863-0618
} In message {199702150110.RAA20593-at-cats-po-1} Jon Krupp writes: } snip } Although we have an oil mist filter } on the VE, we get some smell from the rotary pump when roughing out the } bell jar. Any suggestions on how to eliminate this problem, the computer } users have objected to the smell and possible health concerns. } } Are some filters better than others? How realistic is it to expect a filter } to eliminate all odor? } snip } Any ideas? } } Jonathan Krupp } Microscopy and Imaging Lab } University of California } Santa Cruz, CA 95064 } (408) 459-2477 } FAX (408) 429-0146 } jmkrupp-at-cats.ucsc.edu
I am looking for a person working in materials science and/or physics who would be interested in helping me determine the direction in which to expand our current facilities.
If you are interested, please read on:
I am a biologists who works with light and electron microscopy. I have no idea what else is needed for other fields of study outside of biology.
We have acquired an Hitachi H-7000 with SEM/STEM. We plan to add an EDS system to the scope in order to do elemental analysis. In the process of seeking creative funding (cost sharing with other departments), I proposed the idea of us offering a materials science or physics course to use this instrument.
We are a four-year college with a high caliber student body. We have a small physics department (5 faculty) that the college would like to double in size over the next 5-10 years. As part of a curriculum development group, I proposed that we develop a specialty in physics that typically is not met by other "standard" departments. We currently have an excellent astronomy offering with research-quality telescopes and a planetarium. We also have a well-equipped optics lab. My suggestion was to add a "materials science" component to the department. Not only could we prepare students for graduate work in material science, but we could provide training that would prepare students for direct entry into semi-conductor industry, metallurgy, or whatever.
I currently train a contingent of our students to become microscopists/ cytotechnolgist (we have the TEM, an Hitachi S-510 SEM, vacuum evaporator, two research grade light microscopes with fluorescence, DIC, and image analysis, and complete histology lab, including a cryostat). My idea was to add some breadth to this background by training them in other areas of microscopy (EDS, and other items more relevant to materials science research or industry). Because I am a biologist, I do not know what oher skills this would entail.
And this is where I am asking for help. What else would we want to consider adding? I asked a chemistry colleague about NMR, but apparently the NMR that we have will not do the things that a materials science/physicist would need. It takes the "other kind" of NMR (I believe she said it was a solid state or solid phase NMR that we would need). We have an engineering school in the college. Is there something that would be attractive to them as well?
I am not looking for a place to spend money, but rather am looking for a suggestion of what to include in this proposed program. (I.e., Ideally, what would your include in the range of training for materials science?) It does not have to be entirely microscopy. Our funds are limited, but if we could develop a program/course that could make some of our graduates more useful to industry (particularly industry in New Jersey), there are special grants in the state to support these type of programs.
Please respond with ideas of instruments, their rough cost, and the applications that they would be used for. Please feel free to suggest other forms of training/skills that would be good for us to teach.
Please respond directly to me.
Thank you.
______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 The College of New Jersey fax: (609) 771-2674 Trenton, NJ 08650-4700
Kim, Our Hazardous Waste folks don't like to deal with the substitution fluids we use so check with your waste disposal people before you use the following method:
Make stock solutions of 4% Osmium tetroxide (1g in 25 ml HPLC grade Acetone*) and 0.1% Uranyl acetate (0.025g in 25 ml HPLC grade acetone). Mix stock solutions 1:1 for your substitution fluid. Note: Uranyl acetate takes several hours to go into solution so I make it the night before I need it (keep it in the dark at room temp). Then chill it to -80. *The acetone must be prechilled to -80 before you add the osmium. Keep solutions on dry ice or use a cold block when you bring them out of the -80. Mix the solutions into prechilled sample vials.
References: Hoch HC (1986) Freeze-substitution of fungi. In: Aldrich HC, Todd WJ (eds) Ultrastructure techniques for microorganisms. Plenum, NY
Howard RJ and O'Donnell KL (1987) Freeze substitution of fungi for ctyological analysis. Experimental Mycology 11: 250-269
Best regards, Beth
} Hello all. I know I've seen this topic come up here before, but now that I } need it, I can't find the information. } } Can anyone recommend some solvents, other than alcohols and acetone, } suitable for freeze-substitution. I am having some difficulty obtaining } adequate substitution in some cells of my material (plant bits) and want to } try solvents which may penetrate more effectively. } } Thanks in advance. } } Kim Rensing
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
Two training workshops this summer organised by The Center for Cell Imaging, Yale School of Medicine.
July 3-5: Immunocytochemistry and Cryosections Invited Instructor: Jan W. Slot, Utrecht University, The Netherlands.
A practical course aimed at biomedical researchers interested in learning how to cryosection samples and label them with antibodies. This is a true hands-on workshop with the focus on technique transfer. For this reason registration is limited to 10 participants. Registration fee: $500
July 30- August 2: Microwave Workshop. Biological specimen preparation for TEM using a microwave oven. Hands-on workshop to teach rapid (2-4 hour) specimen processing and embedding. This workshop will also include a demonstration of cryoultramicrotomy and digital image aquisition. Registration fee (including accomodation): $950
All workshops will be held at the Center for Cell Imaging in the Yale School of Medicine.
For more information either contact Paul Webster directly (e-mail : Paul.Webster-at-Yale.edu) or visit our web site: http://info.med.yale.edu/cellimg/CCItraining.html
Best Wishes
Paul Webster. Center for Cell Imaging Yale School of Medicine http://info.med.yale.edu/cellimg
Summer Internship in the Materials Technology Department at Intel Corporation, Santa Clara, California
The TEM group at Intel Corp. in Santa Clara, CA announces a summer internship available for a graduate student in the area of specimen preparation technology. The primary goal of this project will be to help construct an argon ion mill inside the specimen chamber of a field emission scanning electron microscope. The SEM will then be used as an ultra-high precision endpoint detector for controlling the termination of ion milled TEM cross-sections of ULSI semiconductor specimens with very small feature size.
The student should have prior experience in SEM, TEM, and TEM specimen preparation of semiconductor materials, particularly the dimple and ion mill technique.
Requirements: U.S. citizenship or permanent residency, 3.0 gpa or higher, enrolled fulltime in school and able to work full time during the internship.
Intel Corp. is an equal opportunity employer.
Please forward inquiries and resumes to:
Dr. John Mardinly Intel Corporation 3065 Bowers Avenue, Mail Stop SC2-24 Santa Clara, CA 95052-8119
The following summarizes the responses I've received in the past couple of weeks regarding sources of colloidal carbon as an electron dense tracer for vascular leakage:
1. Stephen Edgar at the Univ. of Auckland replied that they have used nuclear track emulsion (NTE) from Amersham to examine capillary functionality in heart. The silver grains were the e-dense tracer. The reference for this is: Choong YS, Gavin JB, Cottier DS, & Edgar SG. (1995) Microvascular incompetence and the failure of hearts to recover contractile function after cardioplegia. European Heart Journal 16(8):1140-1146.
2. Jan Leunissen suggested using ultra small gold/BSA conjugates as "latent" tracers, which are then visualized with silver enhancement on sections. He suggested that 20 nm gold conjugated to BSA would have too great a hydrodynamic radius (with adhering protein plus bound water) to be a suitable tracer.
3. A few respondents also suggested that colloidal carbon would not or does not have sufficient electron density to be a suitable tracer and/or that the particle size distribution would be too heterogenous to make them a suitable tracer. To those respondents I can only refer them to an existing body of literature where colloidal carbon was used for this purpose. I obtained the following references from the MEDLINE database (1986-present):
Beck IT, Morris GP & Buell MG. (1986). Ethanol-induced vascular permeability changes in the jejunal mucosa of the dog. Gastroenterology. 90(5 Pt 1):1137-1145.
Dvorak HF, Nagy JA, Dvorak JT, & Dvorak HM. (1988). Identification and characterization of the blood vessels of solid tumors that are leaky to circulating macromolecules. Am. J Path. 133(1):95-109.
Gouveia MA. (1988). The testes in cadmium intoxication: morphological and vascular aspects. Andrologia. 20(3):225-231.
Gerdes U, Gafvels M, Bergh A & Cajander S. (1992). Localized increases in ovarian vascular permeability and leucocyte accumulation after induced ovulation in rabbits. J Reprod. Fert. 95(2):539-550.
Feng D, Nagy JA, Hipp J, Dvorak HF & Dvorak AM. (1996) Vesiculo-vacuolar organelles and the regulation of venule permeability to macromolecules by vascular permeability factor, histamine, and serotonin. J Exp. Med. 183(5):1981-1986.
I will probably try one or more of the techniques suggested or referenced above and will be happy to share results with those that are interested.
The attached text is an excerpt from a Philips Electron Optics document:
Schottky versus Cold Field Emission
Although Field Emission Guns (FEGs) were already mounted on TEMs a long time ago - the first commercial FEG instrument, the Philips EM400 FEG was launched in 1977 - they did not become very popular until the beginning of the 1990's. This was for a number of reasons. First of all, the early instruments operated only up to 120 kV (by many seen as too low for practical Materials Science work) and suffered from teething troubles (which were all solved eventually). The major problem was, however, related to the type of FEG used (at that time there was only one choice: the Cold FEG). Although the CFEG provides a very high brightness and low energy spread, the total area of the tip emitting electrons is very small, so the total current that can be extracted remains low (somewhere between 10 to 25 nA). This is no problem for small-probe work, where the current is typically around 1 nA, all concentrated in a probe that can be as small as 1 nm (at those currents; with lower currents much smaller probes can be achieved). But it is a severe problem for normal TEM operation. A typical working condition with a strongly diffracting (that is, normal) Materials Science specimen, with a small objective aperture inserted, requires at least 100 nA for convenient operation. The typical working method on the CFEG is therefore usually described as eethe operator having his eye glued to the binocular', with the beam focussed on only a small area of the screen to get enough intensity to work by. That changed in 1990 when Philips Electron Optics introduced the CM20 FEG. Instead of a CFEG this instrument (now superseded by the CM200 FEG) uses a Schottky FEG. The tip of the Schottky FEG has a much larger emitting area than the CFEG (though has very similar high brightness and energy spread), so it is possible to extract a much higher total current out of the tip. In fact, several hundreds of nanoAmp res are easily achieved as emission from the tip. Thus the low-intensity problem no longer exists and nearly 100 Schottky FEG instruments have now been sold, indicating its popularity. Nevertheless persistent rumours have been spread wherein it is stated that the CFEG is superior to the Schottky FEG and misconceptions still exist. This note compares the CFEG with the Schottky FEG and demonstrates that in many respects the Schottky FEG is superior in performance to the CFEG and where it isn't, it is equal.
What are the advantages of field emission over thermionic emission?
In essence what field emission provides is two things: a higher brightness and a lower energy spread. Higher brightness (which is defined as the electrons current emitted per unit area and unit angle of the tip, or A/cm2 srad) translates into two things. For HR-TEM imaging higher brightness means a smaller incidence angle and therefore a much improved spatial coherence. For small-probe work, higher brightness means much more current in a probe of the same size (or still high current in much smaller probes). Lower energy spread gives a much improved temporal coherence for HR-TEM imaging and, of course, a better resolution in Electron Energy-Loss Spectroscopy (EELS). There are also some disadvantages to field emission. These have mainly to do with the fact that FEGs require much better vacuum in the gun, are more expensive to make and thus more expensive to run. Also, due to their much improved performance, FEG instruments are more easily affected by vibrations and stray fields (they are not necessarily more sensitive, it's just that the negative effects of vibrations or stray fields wouldn't be noted on equivalent LaB6 instruments: if you cannot resolve 1 anyway, the fact that it isn't there because of vibrations isn't going to make a lot of difference). The table below gives an overview of the most important characteristics of the difference types of electron emitters, the thermionic emitters tungsten (W) and lanthanum-hexaboride (LaB6) and the CFEG and Schottky FEG. One other type of FEG is frequently mentioned: the thermally stabilised CFEG (TCFEG). It is essentially a CFEG heated to about 1800 K where it will emit more stably than a CFEG. Other than that, it isn't dramatically different from a CFEG and is not considered here any further.
Electron source comparison for TEM Characteristic Tungsten LaB6 CFEG Schottky FEG Brightness {105 ~106 107-109 5x 108 Current in 1 nm spot 0.1 pA 1 pA0 0.1-1 nA 0.5 nA Maximum beam current 1 uA 1uA 10-20 nA } 200 nA Energy spread (lowest) 1.5 eV 0.8 eV 0.3 eV 0.6 eV Energy spread (-at- 3 nA current) 2.0 eV 1.0 eV 0.7 eV 0.7 eV Operating temperature 2700 K 2000 K 300 K 1800 K Requires flashing Never Never Every few hrs Never Current stability (noise/short term) {1% {1% } 5% {1% Current stability (long term) Stable Stable } 10%/hr {1%/hr Life time 60-200 hrs 1000 hrs } 2000 hrs } 2000 hrs Vacuum required (Torr) 10-4 10-6 {-10-10 10-8 to 10-9 Sensitivity to external influence Low Low High Moderate Suitable for conventional TEM Yes Yes No Yes Suitable for nano-analysis No Moderate Excellent Excellent Suitable for HR-TEM Weak Moderate Excellent Excellent
How is field emission achieved?
Field emission takes place when a suitable emitter is placed in an extraction field (it's like the static electricity discharges that you can have when your body approaches something that is charged very differently - except that in the case of field emission, the emission is continuous and not a single bang). In the case of the CFEG, the extraction field is the only cause for emission and the emitter works at room temperature. The tip must be very sharp in order to have emission. In the case of the Schottky FEG, it is not only the field that is important. The tip must also be heated to about 1800 K. Realistically speaking, Schottky emission is therefore more like a combination of field and thermionic emission. The Schottky tip is coated with zirconium-oxide and, with its so-called work function lowered at higher temperature, can achieve emission from much less sharp tips.
Brightness
The brightness of an electron source is the primary performance parameter. It is expressed in A/cm2 srad. The presence of the cm2 (the emitting area) and the srad (the emission angle) in the denominator cause brightness to deviate from what the eye perceives as brightness which is in fact an intensity. Intensity is related to total current and is thus typically high for thermionic emitters. The brightness of these emitter is, however, low because they do emit a lot of electrons but from a very large area and under large emission angles. Perhaps the easiest way to perceive brightness is to relate it to the current in a 1 nm spot. In order to create such small spots for thermionic emitter, we have use a very strong first condenser lens (spot size) and a small condenser aperture. By doing so, we throw away almost all the current emitted (if we included it the spot would become larger) and so end up with very little current. For a FEG we can use a much weaker first condenser lens (the spot is already very small so it doesn't need to get much smaller) and so we only throw away a minor portion and end up with a high current. Although the CFEG in principle can achieve slightly higher brightness than the Schottky FEG, this advantage is commonly lost because the high brightness only exists for a short period after flashing. Under more normal working conditions, the brightness has fallen to that of the Schottky FEG or below. A demonstration of the high brightness achieved by the Schottky FEG is the attainment of a spot of 0.24 nm in size, which had a beam current of 50 pA.
Energy resolution
Ultimately the energy resolution of a CFEG is better than that of the Schottky FEG because it doesn't have such a high operating temperature. In practice, however, the very low resolutions (0.3 eV) are rarely achieved and on very few instruments. They require highly stable guns and high-tension generators, otherwise the inherent small energy spread of the tip is lost through instabilities. Also, when operating under normal HR-TEM imaging conditions, the energy spread is not as low any more because the emission current is higher and both CFEG and Schottky FEG typically have energy spreads of 0.7 to 0.8 eV under those circumstances.
Stability
For some applications such as the standardless EDX analysis normally applied in the TEM, where the total result is time-integrated, the stability of the emission is not important. In other applications, such as scanning or Parallel Electron Energy-Loss Spectroscopy (PEELS) analysis, where several spectra at different energies are recorded to allow collection of data for all relevant elements, unstable emission can range from annoying to unusable (if the PEELS spectra were taken under different currents, one cannot integrate the data together into a single analysis). Much of the short-term (in)stability of the CFEG is caused by the effects of molecules from the vacuum attaching temporarily to the tip, then evaporating again. Since the emitting area is so small, a single molecule can affect the emission by several percent. The typical changes in emission of the CFEG are around 10%. As a consequence, CFEG are usually notorious for their flickering images (sometimes so bad that TV-rate cameras become almost unusable because they try to correct for the changes in intensity). In scanning this typically results in horizontal streaks or bands of varying intensity that disfigure the images. The long-term stability of the CFEG is also poor, once again due to the adhesion of molecules from the vacuum. Over time the evaporation of these molecules is less than the adhesion so over a period of hours the emission tends to go down. A typical behaviour of a CFEG is a rapid decline in emission just after flashing (for a description of flashing, see further below), then a period of up to a few hours of stable emission and then a continuous further decline (at which point one typically flashes the tip again). In the case of the Schottky FEG short-term and long-term stability are much higher. Even though the vacuum around the Schottky emitter is normally at least a factor 10 worse than what is acceptable for a CFEG, the emission is very stable. In part this is because the tip appears to be self-cleaning and doesn't show any degradation over time, so the long-term stability of the tip is around 1% change in emission per hour. The much better short-term stability derives partly from the self-cleaning character of the tip and partly from the fact that the emitting area is much larger than that of the CFEG so adhesion of a molecule has much less of an effect percentagewise. As a consequence the Schottky FEG is far superior to the CFEG in applications where beam-current stability is important.
Source size and spot size
A common source of confusion is the fact that the virtual source size of the CFEG is around 3 nm and that of the Schottky FEG is around 20 nm. Often this is misconstrued to mean that the CFEG can generate smaller spot sizes. Nothing is further from the truth. In fact, source size and spot size are not directly related. This is very easy to realise from the following. On a typical LaB6 filament the source size is several micrometres. Yet TEMs equipped with such filaments are capable of achieving spot sizes of 1 nm or smaller. Evidently the condenser-lens system of the microscope can reduce the source size (called demagnifying) from say 2 m down to 1 nm, a reduction of 2000x. The FEG instruments are similarly equipped with a condenser system. Thus going from a virtual source size of 20 nm to a spot size of 0.2 nm is only a reduction of 100x. In fact, the condenser system of the CM200 FEG is used only over part of its range in comparison with the equivalent LaB6 microscopes. Incidentally, source size has very little to do with the emitting area. In FEGs the source size is called virtual because the electron trajectories from tip appear to emanate from a small area inside the tip itself. Thus the real emission area on a CFEG is larger than 3 nm and that of the Schottky FEG also larger than 20 nm (in fact it is several hundreds of nanometres is diameter). Thermionic filaments in contrast do not have a virtual source size. In their case the source is the size of the first cross-over after the filament.
Maintenance
In the case of the Schottky emitter, maintenance is quite a bit easier than for a CFEG. The vacuum in the gun doesn't need to be as good as for a CFEG. The baking of the gun after installing a new tip is therefore much shorter (typical tip exchange times are just a few days, including 12 to 24 hours of baking of the gun). This contrasts with at least several days of gun baking for a CFEG. Also, since the vacuum is less critical, the construction of the vacuum system is easier, making maintenance easier and more rapid.
Flashing
Finally, what is flashing? Flashing is heating the tip of the CFEG up to very close to its melting point. This causes the evaporation of all molecules that were deposited. Also the tip reshapes itself slightly (normally this is benign; however, sometimes the reshaping goes wrong and the tip becomes too blunt for good field emission so it must be changed). Although baking itself takes only a few minutes, the gun is usually quite unstable after that, with a lot of drift of the gun alignment, so typically one half hour goes by in which it is difficult to work. As mentioned before, the Schottky FEG tip is self-cleaning. It is therefore is never flashed. In the morning you turn up the extraction voltage to the value where you want to work and it will emit stably. And at night the last user can switch the gun to standby. And in between it can be used continuously and with very high stability.
Conclusion
The CFEG has a few minor advantages relative to the Schottky FEG but in most cases these do not adequately compensate the CFEG's deficiencies. As a result the Schottky FEG is the field emitter of choice over the whole range of TEM techniques, from eenormal' TEM imaging, HR-TEM imaging, small-probe analysis, diffraction to scanning.
Dr. Max T. Otten TEM Application Software Manager / TEM Application Specialist Philips Electron Optics Applications Laboratory Bldg. AAE 5600 MD Eindhoven The Netherlands tel. +31-40-2766106 fax +31-40-2766102 ________________________________________________________
I had trouble coating gold particles with ricin, a plant lectin. After centrifugation the pellet contained mostly aggregates not usable for further experiments. I would appreciate some advice on which pH to choose for conjugation and if one should add lactose in order to blokk binding sites of ricin. Any helpful comments are greatly appreciated.
Thanks Andreas
Andreas Brech Electron Microscopical Unit for Biological Sciences Department of Biology, University of Oslo. P.O.Box 1062 Blindern N-0316 Oslo 3 Norway Tel.: + 47-22 85 61 89 (work) + 47-22 43 83 23 (privat) Fax.: + 47-22 85 47 26 e-mail.: abrech-at-bio.uio.no
Dear Don, I run an EM lab in the Metals and Materials Engineering department at UBC and by far the most useful equipment for Materials Engineers, all Engineers and Physicists is the combination of SEM and EDX. I have two SEM+EDX's and one TEM with STEM, SEM and EDX. Specimen preparation for materials TEM is complicated and very different from biological TEM, although some ultramicrotomy is used for specialized techniques. You must use electro-polishing with explosive acid mixtures with currents running through them to thin metals. Most of the techniques are black art and you need an expert to teach you. The non-conducting samples (ceramics, rocks, semi-conductors) or multi-phase metals must be thinned with an ion-beam thinner (~$50,000US). First the 3 mm. sample is cut with a slurry drill (~$500), then thinned to about 100 microns with a disc-thinner (~$100), then dimpled with a Dimpler (~$15,000), then ion-beam-thinned to perforation. My first recommendation would be a good light-element EDX for your SEM and a course on quantitative analysis. This will satisfy most of your materials EM requirements. Next would be a back-scattered detector. You must carbon-coat all non-conductors for EDX. Epoxy-mounting capacities and a grinding and polishing setup will be necessary, but the engineering or geology departments may have this if they do light microscopy. Always use diamond final polish for EDX. Other things on a wish list would be: FEG SEM, AFM, SIMS. It depends on the direction of your engineering and physics departments. You wrote:
} I am looking for a person working in materials science and/or physics who } would be interested in helping me determine the direction in which to } expand our current facilities. ...snip } And this is where I am asking for help. What else would we want to } consider adding? I asked a chemistry colleague about NMR, but apparently } the NMR that we have will not do the things that a materials } science/physicist would need. ..snip } I am not looking for a place to spend money, but rather am looking for a } suggestion of what to include in this proposed program. (I.e., Ideally, } what would your include in the range of training for materials science?) } It does not have to be entirely microscopy. Our funds are limited, but if } we could develop a program/course that could make some of our graduates } more useful to industry (particularly industry in New Jersey), there are } special grants in the state to support these type of programs. } } Please respond with ideas of instruments, their rough cost, and the } applications that they would be used for. Please feel free to suggest } other forms of training/skills that would be good for us to teach. This is just a quick overview of the emphasis of materials EM. ("Tell me all you know in 25 words or less"). Please feel free to contact me for more specific questions. Regards, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
Dear All, I have read that paper by Phillips about cold and thermal FEG, but when I had a deomonstration of a cold FEG, 200 kV TEM (Hitachi HF-2000) at the Seattle ICEM '90, it worked just fine for conventioal TEM on a show floor, with bright overhead lights and some interference from the security guards' walkie-talkies. We did low-mag, high-mag and roamed all over the lattice images of the crystal. I never used the binoculars and the TV camera was showing everyone else what I was seeing on the screen. I also saw a EELS spectra that showed 0.1ev energy spread, as measured as the FWHM of the zero-loss peak. Don't believe everything the company man tells you. Regards, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
"I had trouble coating gold particles with ricin, a plant lectin. After centrifugation the pellet contained mostly aggregates not usable for further experiments. I would appreciate some advice on which pH to choose for conjugation and if one should add lactose in order to blokk binding sites of ricin. Any helpful comments are greatly appreciated."
Dear Andreas, If you really need to couple the ricin to the gold then there are a few things to look out for. Firstly, the protein is best coupled at a pH whose value is just lower than the isoelectric point of the protein (there is a study by Horisberger and Clerc somewhere in the literature on this).
Secondly, the coupling of the gold and protein can be performed in either a weak salt solution or even water if the ricin is stable in this medium.
Fianlly, if there is no aggregation prior to centrifugation and the correct amount of protein has been added (determined by inhibition of electrolyte-induced aggregation), your problem with aggregation may be a result of a too high centrifugation speed. Try spinning the preparations at slower speeds to see if you can stop the aggregates from forming.
However, if your wish is to localize ricin on tissue sections and the conjugation process is not working, then why not try looking for unconjugated, bound ricin on your samples using anti-ricin antibodies? This may be a much simpler solution to your problem.
Best regards,
Paul Webster Center for Cell Imaging Yale School of Medicine http://info.med.yale.edu/cellimg
Visit our colloidal gold pages at http://info.med.yale.edu/cellimg/gold.html
"I had trouble coating gold particles with ricin, a plant lectin. After centrifugation the pellet contained mostly aggregates not usable for further experiments. I would appreciate some advice on which pH to choose for conjugation and if one should add lactose in order to blokk binding sites of ricin. Any helpful comments are greatly appreciated."
Dear Andreas, If you really need to couple the ricin to the gold then there are a few things to look out for. Firstly, the protein is best coupled at a pH whose value is just lower than the isoelectric point of the protein (there is a study by Horisberger and Clerc somewhere in the literature on this).
Secondly, the coupling of the gold and protein can be performed in either a weak salt solution or even water if the ricin is stable in this medium.
Fianlly, if there is no aggregation prior to centrifugation and the correct amount of protein has been added (determined by inhibition of electrolyte-induced aggregation), your problem with aggregation may be a result of a too high centrifugation speed. Try spinning the preparations at slower speeds to see if you can stop the aggregates from forming.
However, if your wish is to localize ricin on tissue sections and the conjugation process is not working, then why not try looking for unconjugated, bound ricin on your samples using anti-ricin antibodies? This may be a much simpler solution to your problem.
Best regards,
Paul Webster Center for Cell Imaging Yale School of Medicine http://info.med.yale.edu/cellimg
Visit our colloidal gold pages at http://info.med.yale.edu/cellimg/gold.html
} Date: Sat, 22 Feb 1997 10:46:11 +0100 } From: Andreas Brech {andreas.brech-at-bio.uio.no} } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: colloidal gold } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hi } } I had trouble coating gold particles with ricin, a plant lectin. After } centrifugation the pellet contained mostly aggregates not usable for further } experiments. I would appreciate some advice on which pH to choose for } conjugation and if one should add lactose in order to blokk binding sites of } ricin. Any helpful comments are greatly appreciated. } } Thanks Andreas } } Hope to be able to help
First you should adjust the Ph of the colloidal gold to the exact PI(isoelectric point) of your lectin. Very important, also you should add the minimum amount of lectin that will allow a good stable complex. Once the protein or lectin is added you can try the Nacl test on a few drops of the complex before centrifugation, the solution should not change colour, then the complex is probably stable.
Good luck Sylvia
} } Andreas Brech } Electron Microscopical Unit for Biological Sciences } Department of Biology, University of Oslo. } P.O.Box 1062 Blindern } N-0316 Oslo 3 } Norway } Tel.: + 47-22 85 61 89 (work) } + 47-22 43 83 23 (privat) } Fax.: + 47-22 85 47 26 } e-mail.: abrech-at-bio.uio.no }
A collegue (entomolgist) has asked for good textbooks on preparation of arthropods for a) Histology, and b) SEM. I'm presuming he's looking at the former for soft tissues and the latter for skeletal morphology. All I have is lots of bits and pieces from the Methods sections of papers. Does anyone have any suggestions?
Many thanks,
Geoff Avern Manager Microscopy Labs Australian Museum Sydney, Australia
We have been recently trying Picric Acid in some of our primary fixative recipes along with Glutaraldehyde and Paraformaldehyde (with Brain Tissue). We have been experiencing some precipitation on the membranes in the sections and we have established that the problem isn't through section staining.
Having done some reading we have found it stated in one EM textbook that using Picric acid in the primary fix and then doing buffer washes (any buffer or any aqueous solution) will cause insoluble picrates to be deposited. It is suggested in this text that the tissue should be placed directly into 50% EtOH to dissolve away the picrates and to wash away the glut. (Text; Resin Microscopy and on-section Immunocytochemistry pp 57).
Has anyone got any comments about the use of Picric Acid in the primary fixative and in particular the problem of buffer washes after the primary fixation (which includes Picric Acid) causing precipitation?
If we are to osmicate after primary fix, what should the washes between each step be in ?
I would appreciate any comments regarding this query,
TIA
Rich.
----------------------------------------------------------------------- Richard Lander, NZCS South Campus Electron Microscope Unit c/- Pathology Department Otago Medical School P.O. Box 913 Dunedin New Zealand. Tel. National 03 479 7301 Fax. National 03 479 7254
"Southernmost EM Unit in the World!" ------------------------------------------------------------------------
Geoff Avern wrote: ====================================================== A collegue (entomolgist) has asked for good textbooks on preparation of arthropods for a) Histology, and b) SEM. I'm presuming he's looking at the former for soft tissues and the latter for skeletal morphology. All I have is lots of bits and pieces from the Methods sections of papers. Does anyone have any suggestions? ====================================================== The book Scanning Electron Microscopy of Medically Important Arthropods by Viqar Zaman, Ph. D., 1983, 175 pages while a bit dated is still one of the best books I have seen on this subject. Maybe there are others but Prof. Zaman was quite an experimentalist and developed quite a few novel methods of preparation. The last I heard he was still teaching and doing research in Pakistan.
Disclaimer: SPI offers this book in its library section!
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
------------------------------------- I have been hunting the net to get info on the old NOVA Demon-Plus systems. Any suggestions? I'm busy developing user-friendly software for converting AN10000 to PC and need this info. Craig
Snip=8A"Don't believe everything the company man tells you."
Right, but=8A
1. We are the owner and a user of both a Hitachi HF-2000 cold FEG TEM/STEM and a Philips 300 kV FEG/Schottky. As such I can tell you that you will not get 1 meV energy spread on an EELS spectra on the Hitachi HF-2000 under practical condition for microanalysis if ever.
2. Moreover, you will not be able to work "with bright overhead lights" in low mag on our instrument.
To my opinion, the gun brightness is not the only point to consider. The size of the apertures in the illumination section of the microscope is also very important, in particular the presence of fixed apertures that may be added to achieve thin and clean probes or STEM resolution and may dramaticaly reduce the intensity. I am wondering if the instrument she have seen in Seattle didn't have a special set up for exhibition which may be different from the research one (customer configuration).
Summer Internship in the Materials Technology Department at Intel Corporation, Santa Clara, California
The TEM group at Intel Corp. in Santa Clara, CA announces a summer internship available for a graduate student in the area of specimen preparation technology. The primary goal of this project will be to help construct an argon ion mill inside the specimen chamber of a field emission scanning electron microscope. The SEM will then be used as an ultra-high precision endpoint detector for controlling the termination of ion milled TEM cross-sections of ULSI semiconductor specimens with very small feature size.
The student should have prior experience in SEM, TEM, and TEM specimen preparation of semiconductor materials, particularly the dimple and ion mill technique.
Requirements: U.S. citizenship or permanent residency, 3.0 gpa or higher, enrolled fulltime in school and able to work full time during the internship.
Intel Corp. is an equal opportunity employer.
Please forward inquiries and resumes to:
Dr. John Mardinly Intel Corporation 3065 Bowers Avenue, Mail Stop SC2-24 Santa Clara, CA 95052-8119
We are desperately needing to buy two used heads (chuck holder which fits into microtome cutting arm) for our aged Ultracut E ultramicrotomes. Does anyone have any lying around not being used?
We have been experienceing a 50% loss of Epon-Araldite embedded thin sections from nickel grids during immunostaining (Au). We have tried coating mesh grids with dilute formvar or butvar solutions to no avail. Can anyone help?
A colleague, Dr. Goverdina Fahraeus-Van Ree, is in urgent need of 25-50 g of low viscosity PolyPep P5115 manufactured by Sigma.
It has been back-ordered and will not be shipped before she needs it. The item is a mixture of polypeptides used as a stabilizer for histochemistry work. If you have some you could 'loan' her and have it replaced when her order arrives, could you email her directlyy at gvanree-at-morgan.ucs.mun.ca
Thanks
Carolyn J. Emerson email: cemerson-at-plato.ucs.mun.ca
Biology Department Memorial University St. John's, NF A1B 3X9 Tel: (709) 737-7515 Fax: (709) 737-3018
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Question: are you using Tween in your washing steps? We had the same problem and came to the conclusion that there was too much Tween. It is very viscous and can cling to the outside of the pipette tip increasing the concentration then acting like washing-up liquid, making squeaky clean grids! Try a grid without and see what happens. Elaine
Dr. Elaine Humphrey Biosciences Electron Microscopy Facility University of British Columbia 6270 University Blvd Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-unixg.ubc.ca
At 11:11 AM 24-02-97 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Put 100 ml of chloroform in a brown bottle (reduces photolysis) and somehow poke the tape down the bottleneck. The tape should not change in appearance but the glue dissolves into the chloroform if you leave it for 48 hrs. If the tape goes milky of shrivels up try a different kind. You dont want to use tape solution, just glue.
The grids as supplied are hydrophobic. I make them hydrophilic by whisking them through a small (spirit lamp) flame. They flash red and change colour. If they shrivel, whisk faster. Place flamed grids on filter paper, put a drop of glue in chloroform on each grid, leave to dry, use right away. OR dip each grid in the glue bottle.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} I am wondering if the instrument she have } seen in Seattle didn't have a special set up for exhibition which may be } different from the research one (customer configuration). } } "Don't believe everything microscopists tells you!" } } Best regards } } Philippe-A. Buffat
I would add my voice to that caution. I've worked as a demonstrator for EM manufacturers, and while what you see at an exhibition is very unlikely to have been specially 'modified' - I've certainly never done it, and have never heard of it being done - a skilled demonstrator can operate an instrument to show it in the best possible light, and very quickly!
In the example quoted, I would guess the demonstrator was doing some quick work with condenser apertures. The lesson is to use what you see at an exhibition as a taster for what might be possible. If you have a serious interest, you need a proper demonstration where you can really check how the instrument operates.
I am involved in a project which requires the localization of an antibody made by a dog with an autoimmune disease. Due to the density of the structure to which this antibody is likely to bind, I would like to first diffuse the antibody thru the tissue to the site of binding, then to follow it with the diffusion of a 1-nm gold secondary conjugate so that it may be localized in the EM after silver enhancement. I have not been able to find a canine-specific 1 nm gold secondary conjugate. I understand, though not thru personal experience, that protein-A binds canine antibody with high affinity. My question is: Has anyone had a favorable experience with a particular brand of a PA-1nm gold conjugate? I just wasted a week of time using a defective conjugate from a manufacturer with whom I had no prior experience, and hope not to fall into the same trap again. Better yet, does anyone know of a manufacturer of a anti-dog 1 nm conjugate?
Many thanks in advance!
Doug Keene Portland Research Unit Shriners Hospital for Children
Dear all, We have JEM4000 and JEM2010 here and now are going to change from dark room to digital imaging. The problem is (if forget about funding:) to choose what to buy. Reading manufactures information is not the best way to make the choice because all are the best:). Would the people who directly work with digital imaging express their opinion of different types of apparatus and soft? Here are topics I'm exactly interested in but any user information will be welcomed: 1. CCD cameras for HR low dose work (manufacturers, properties etc.) 2. Solutions to combine high quality slow scan images with fast scan (} =5fr/sec) to search dynamics. 3. PC interfaces (or strong arguments for other platform), convenience in use, possibilities, online loop-back. 4. Software to process and archive images. 5. Storing media (MO disks, CD writable, DVD(?), strimmers, etc.(?)). 6. Output devices (dye sublimation vs. laser-printers, other(?)). 7. Did anyone make comparative cost estimations for digital imaging vs. dark room?
I hope the discussion will be of common interest. TIA Andrew Chuvilin Siberian Center for Electron Microscopy Novosibirsk Russia
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Subject: Time:8:30 AM OFFICE MEMO etch Date:2/25/97
Does anyone have recipes for "etching" the surface of LR White in order to expose antigenic sites for surface immunogold locaizations? Also, a recipe or reference to the procedure for etching Epon or Araldite thin sections for immunogold. We have used it several years ago but perhaps there is a better method than NaMethoxide. thanks jim jamieson
I would like to inquire if anyone has had experiences, favorable or unfavorable, with color CCD cameras for light microscopy. I would like to purchase one for recording pictures from histology slides but recognize a wider range of applications.
Are there any that are particularly good buys, easy to interface with a host computer, easy to use? Any that are nightmares?
Anyone have any experience with Microlumina that they would like to pass on?
Kenneth A. Taylor, Ph.D. Phone: 904-644-3357 Institute of Molecular Biophysics Fax: 904-561-1406 Florida State University E-mail: taylor-at-sb.fsu.edu Tallahassee, FL 32306-3015 Home pages: http://www.sb.fsu.edu/~taylor/ http://www.fsu.edu/~biology/faculty/kat.html
} A collegue (entomolgist) has asked for good textbooks on preparation of } arthropods for a) Histology, and b) SEM. I'm presuming he's looking at the } former for soft tissues and the latter for skeletal morphology. All I have } is lots of bits and pieces from the Methods sections of papers. Does } anyone have any suggestions? } } Many thanks, } Geoff, You already have the best references. Arthropod histological methods are mostly scattered through the primary literature, and in graduate theses. General histo-technique books, such as Kiernan's _Histological and Histochemical Methods_ are as good as general arthropod books. Every group and structure will have its own problems; the solutions would be best found in the lab, or the primary literature. But there are some general sources, mostly as histology references. I haven't seen the book Garber recommended, but it sounds interesting. Some others to check: For methods-- _Histology of the Blue Crab, _Callinectes sapidus_: A Model for the Decapoda_, 1980, by Phyllis T. Johnson pub. by Praeger. _Zooplankton Fixation and Preservation_, Monographs on Oceanographic Methodology 4, 1976, H. F. Steedman, ed., The UNESCO press. _A Colour Atlas of Insect Tissues via the Flea_,1986, Miriam Rothschild, Yosef Schlein, and Susumo Ito, Wolfe Publishing Ltd. For histology--- _Comparative Animal Cytology & Histology_ trans. 1976, Ulrich Welsch and Volker Storch. U. Washington Press. _Microscopic Anatomy of Invertebrates_, vol. 9 _Crustacea_ and vol. 10 _Decapod Crustacea, both 1992, Frederick W. Harrison and Arthur G. Humes, eds. Wiley-Liss (Jon Wiley & Sons). _Biology of the Arthropod Cuticle_ Zoophysiology and Ecology 4/5, 1975, A.C. Neville, Springer-Verlag. SEM techniques may also be found in David Scharf's books of SEMs, and the various coffee-table books of SEMs. The range of SEM methods is very broad--I've used everything from just ripping off the bit I was interested in (yes, the critters were dead), and throwing it in the SEM unprocessed and uncoated, to live animals, to formally fixed, dehydrated, and dried samples. It all depends on what you want. The only general hint is that arthropod cuticle takes up osmium very poorly, but OsO4 can still be useful for SEM by providing greater conductivity and reducing charging, especially of setae. I found that *in general*, this can take 4% OsO4 for 4 or more hours. And don't use a phosphate buffer, or there will be precipitates formed. Phil
&&& Illigitimi non carborundum &&&&&&&& Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217)355-1143 evenings oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
} Date: Mon, 24 Feb 1997 12:32:28 -0800 (PST) } From: Elaine Humphrey {ech-at-unixg.ubc.ca} } To: HILDEGARD CROWLEY {hcrowley-at-odin.cair.du.edu} } Cc: Microscopy-at-Sparc5.Microscopy.Com } Subject: Re: TEM:Help! Loosing immuno thin sections from grids } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } -----------------------------------------------------------------------. } } } } We have been experienceing a 50% loss of Epon-Araldite embedded thin } } sections from nickel grids during immunostaining (Au). We have tried } } coating mesh grids with dilute formvar or butvar solutions to no avail. } } Can anyone help? } } Question: are you using Tween in your washing steps? } We had the same problem and came to the conclusion that there was too much } Tween. It is very viscous and can cling to the outside of the pipette tip } increasing the concentration then acting like washing-up liquid, making } squeaky clean grids! Try a grid without and see what happens. } Elaine } } Dr. Elaine Humphrey } Biosciences Electron Microscopy Facility } University of British Columbia } 6270 University Blvd } Vancouver, BC } CANADA, V6T 1Z4 } Phone: 604-822-3354 } FAX: 604-822-6089 } e-mail: ech-at-unixg.ubc.ca } Are you putting the sections on the dull side of the grid? (They should be.)
Another thing to try is to dip the grids in 10% acetic acid, and then rinse in water before collecting the sections. 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
If you can't find any Ultracut E specimen holders, you might try contacting RMC.
I no longer work there, but if memory serves me right there are some versions of RMC specimen holders that are interchangeable with Ultracut holders.
Anyway, it's worth a try. Contact RMC at: RMCBTLI-at-aol.com or call them in the U.S. at: (520) 889-7900. Your contact person there is Dr. Greg Becker.
The mailing list is Confocal Microscopy List {CONFOCAL-at-UBVM.CC.BUFFALO.EDU} but don't use that to subscribe. Instead, send a message to: listserv-at-ubvm.cc.buffalo.edu
on the first line of the message (ignore subject) type:
subscribe confocal Your Full Name
the subscription must be carried out from your account because the listserver will parse the address from your header and you will be added to the list.
Here's a confocal web page I ran across while searching my old mail for the above, I haven't looked at it recently.
We've recently bought the MicroLumina but don't have it running yet. Silly me thought I'd go with the best computer I could afford, which included Win NT v4.0. Unfortunately, neither the Leaf MicroLumina nor the Primera Pro dye sub printer have drivers for NT v4.0 yet. Looks like I'll have to wipe NT v4.0 and install Win 95 (for joy!*#-at-!!) until the drivers come along. This will upset our network administrator!
Lesson: whatever you consider, check for compatibility with OS's (and with your net admin, too).
Geoff Avern
Microscopy Labs Australian Museum Sydney, Australia
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I would like to inquire if anyone has had experiences, favorable or unfavorable, with color CCD cameras for light microscopy. I would like to purchase one for recording pictures from histology slides but recognize a wider range of applications.
Are there any that are particularly good buys, easy to interface with a host computer, easy to use? Any that are nightmares?
Anyone have any experience with Microlumina that they would like to pass on?
Kenneth A. Taylor, Ph.D. Phone: 904-644-3357 Institute of Molecular Biophysics Fax: 904-561-1406 Florida State University E-mail: taylor-at-sb.fsu.edu Tallahassee, FL 32306-3015 Home pages: http://www.sb.fsu.edu/~taylor/ http://www.fsu.edu/~biology/faculty/kat.html
} I am involved in a project which requires the localization } of an antibody made by a dog with an autoimmune } disease. Due to the density of the structure to which this } antibody is likely to bind, I would like to first diffuse } the antibody thru the tissue to the site of } binding, then to follow it with the diffusion of a 1-nm } gold secondary conjugate so that it may be localized in the } EM after silver enhancement.
I can't help with an anti dog, but I have had experience diffusing antibody through tissue (fixed). I found that even 1nm gold-antibody complex didn't get in and had to turn to 1nm gold conjugated to Fab fragments. And saponin in all solutions was a must. Good luck and Email me if you want to.
Diana van Driel Dept Ophthalmology Sydney University C09 AUSTRALIA 2006
A quick question to those involved in multi-user environments,
We are currently reviewing our policy on access and useage to our multi-user EM unit. The issue of security came up. What we are wanting to find out from various people in multi-user situations, what precautions are taken for security in the lab.
In particular, we (staff of the unit) may be in the TEM/SEM/darkroom/prep area, which are all away from the main door. The question is, how can we prevent someone coming into the office area, and taking off with stuff?
Are people keeping the door locked and only allowing users with keys in, or are 'swipe cards' used, or combination locks of some sort used? At the moment, our users are issued with a key, which generally they have to use when we are out of the lab (lunch, smoko, home). Should we be locking the door while we are IN the lab?
What has prompted this sort of query, is that in the past few months there has been some thefts in a dept along from ours, which operates on a similar system.
We would be interested to hear from all those in multi-user situations as their policies/ideas on security.
Thanks for your time,
Rich Lander.
----------------------------------------------------------------------- Richard Lander, NZCS South Campus Electron Microscope Unit c/- Pathology Department Otago Medical School P.O. Box 913 Dunedin New Zealand. Tel. National 03 479 7301 Fax. National 03 479 7254
"Southernmost EM Unit in the World!" ------------------------------------------------------------------------
Reply to Richard Lander's problem with picric acid precipiatates in tissues:
I have used picric acid in my primary fixative for many years. The fixative I use is 3% paraformaldehyde, 3% glutaraldehyde in cacodylate buffer with 0.1% picric acid. I have also used phosphate buffer to get away from the arsenic. After making up the fixative I always filter it using a 0.2 micron in-line filter.
This fixative is used routinely in whole body perfusions for neurotoxic studies but I have used it as a routine fix. I usually process the peripheral nerves, dorsal and ventral roots from the spinal cord both cervical and lumbar in plastic and the brain and spinal cord in paraffin without any problems with precipitates.
Cheryl Rehfeld Manager Anatomic Pathology Texas Children's Hospital Department of Pathology Houston, TX ---------- -----------------------------------------------------------------------.
Hi there,
We have been recently trying Picric Acid in some of our primary fixative recipes along with Glutaraldehyde and Paraformaldehyde (with Brain Tissue). We have been experiencing some precipitation on the membranes in the sections and we have established that the problem isn't through section staining.
Having done some reading we have found it stated in one EM textbook that using Picric acid in the primary fix and then doing buffer washes (any buffer or any aqueous solution) will cause insoluble picrates to be deposited. It is suggested in this text that the tissue should be placed directly into 50% EtOH to dissolve away the picrates and to wash away the glut. (Text; Resin Microscopy and on-section Immunocytochemistry pp 57).
Has anyone got any comments about the use of Picric Acid in the primary fixative and in particular the problem of buffer washes after the primary fixation (which includes Picric Acid) causing precipitation?
If we are to osmicate after primary fix, what should the washes between each step be in ?
I would appreciate any comments regarding this query,
TIA
Rich.
----------------------------------------------------------------------- Richard Lander, NZCS South Campus Electron Microscope Unit c/- Pathology Department Otago Medical School P.O. Box 913 Dunedin New Zealand. Tel. National 03 479 7301 Fax. National 03 479 7254
"Southernmost EM Unit in the World!" ------------------------------------------------------------------------
} I want to ask if anyone works paraffin shrimp processing and microtomy. } I have troubles in shrimp paraffin sectioning, because of the hard } skeleton of the shrimp. } How could I soften the exoskeleton of the body of the shrimp ?. } How could I soften the chitinous exoskeleton of the shrimp to make paraffin } sections ?. } racosta-at-ccr.dsi.uanl.mx
You will have the best luck if you plastic embed. If you need to use paraffin, use the hardest grade that you can get. If the cuticle is heavily calcified, decalcify with EDTA, unbuffered formalin, or some of the gentler decalcifying methods (which I forget--this was addressed recently in this group). A trick if you're using paraffin is to put some dry ice on top of the block to get it cold, then cut. You won't get ribbons, but you might get sections. Phil
&&& Illigitimi non carborundum &&&&&&&& Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217)355-1143 evenings oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
I just had a demonstration of the Pixera Professional color camera and, although it is not as nice as a true 3CCD RGB camera, for the price (approx. $1200 US) it seems like a good buy. Our lab is also interested in histology slides; we want not only to record them but to quantify image features. I found the Pixera provided better resolution than our existing 1chip CCD B/W camera, even when using the 640x480 resolution mode. But it will go up to 1260x960 (very slow). It is not real time (achieves resolution by combining frames somehow), and the color image files don't always behave like 24 bit true color, but visually they are quite nice. I have been able to open them in Adobe Photoshop and separate out colored features (yellow vs. red) to create different grey images for thresholding and analysis. This camera does require a computer for its card, and (I think) one does not have the option of a video signal.
I am thinking of buying one of these myself. What do others think of this camera??
-Karen P.S. I have no connections with Pixera other than that of a potential customer
Kenneth Taylor wrote: } } I would like to inquire if anyone has had experiences, favorable or } unfavorable, with color CCD cameras for light microscopy. I would like to } purchase one for recording pictures from histology slides but recognize a } wider range of applications. } } Are there any that are particularly good buys, easy to interface with a } host computer, easy to use? Any that are nightmares? } } Anyone have any experience with Microlumina that they would like to pass on? } } Thanks for any insight you care to pass on. } } Cheers -- Ken } } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { } } Kenneth A. Taylor, Ph.D. Phone: 904-644-3357 } Institute of Molecular Biophysics Fax: 904-561-1406 } Florida State University E-mail: taylor-at-sb.fsu.edu } Tallahassee, FL 32306-3015 } Home pages: http://www.sb.fsu.edu/~taylor/ } http://www.fsu.edu/~biology/faculty/kat.html } } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { {} } { } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Life Sciences Sector Lab Reply: kszaruba-at-mmm.com 3M Company 3M Center 270-1S-01 Phone: 612-737-2971 St. Paul, MN 55144-1000 Fax: 736-1519
These opinions are my own and may not represent those of 3M.
Have a look in our links, there is a section on confocal microscopy and another section "Societies and Fora" which has a link to the confocal server. Currently they still have a message up "Seasons Greetings"; perhaps that is for Easter. Jim Darley
ProSciTech Microscopy Supplies & Accessories PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, links, MSDS ************************ http://www.proscitech.com.au
---------- } From: Anja Nenninger {nenning-at-uft.uni-bremen.de} } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: confocal laser scanning microscopy } Date: Wednesday, 26 February 1997 3:16 } } -----------------------------------------------------------------------. } } Dear microscopists, } } Does anyone know wether there is a special listserver for confocal laser } scanning microscopy? } } A. Nenninger } University of Bremen } Physiological Plant Anatomy } Workgroup Heyser } Leobener Strasse UFT } D-28359 Bremen } Nenning-at-uft.uni-bremen.de } Phone: +49/421-218-2954 }
I have a SEM machine , model Cambridge Stereoscan100 and trying to get a frame grabber with a digital imaging analysis software. I look around for a video output from the machine but couldn't find any. Just what type of signal they are using ( Pal, NTSC, SECAM.etc) and where do I need to get that signal. This is a very old machine and I wonder if anyone out there have any experience in doing so?
Reply-To: leeman-at-VOEDING.TNO.NL Transduction Laboratories sells anti-E-cadherin In the Netherlands the distributors are: Braunschwig chemie and Thamer Diagnostica. This antibody has been referenced in papers since 1988! Larry D. Ackerman (415) 476-8751 Howard Hughes Medical Institute FAX (415) 476-5774 UCSF, Box 0724, Rm U426 533 Parnassus Ave. mishot-at-itsa.ucsf.edu San Francisco, CA 94143
I am trying to compile a list of what type of disposable gloves should be used with what chemicals found in our EM unit. I have compared information from various sources and there seems to be a lack of clear data for some chemicals (although no shortage of opinions from some users). As a result of this survey I have got an idea of what is the general consensus for each chemical, this appears below. I would appreciate any comments, especially if anyone considers me to be definately 'off the mark' on any item:
Some, as you can see, I haven't been able to decide on.I want to limit the gloves to disposables where possible.
Chemical:Glove polymer type (latex, neoprene, nitrile, PVA or polyethylene)
acrylic resins=DD (Lowicryl, LR Gold, Unicryl): Neoprene, nitrile(?)
Thanks in advance to anyone who bothers reading this and responding!
Richard Easingwood
----------------------------------------------------------------------- Richard Lander, NZCS South Campus Electron Microscope Unit c/- Pathology Department Otago Medical School P.O. Box 913 Dunedin New Zealand. Tel. National 03 479 7301 Fax. National 03 479 7254
"Southernmost EM Unit in the World!" ------------------------------------------------------------------------
In a message dated 97-02-26 03:14:35 EST, hhlim-at-qes.po.my (Lim Hian Ho) writes:
} } I have a SEM machine , model Cambridge Stereoscan100 and trying to get a } frame grabber with a digital imaging analysis software. I look around for a } video output from the machine but couldn't find any. Just what type of } signal they are using ( Pal, NTSC, SECAM.etc) and where do I need to get } that signal. This is a very old machine and I wonder if anyone out there } have any experience in doing so?
There's more to digital imaing than attaching a frame grabber to the EM. I dont believe Cambridge supports this machine anylonger, but a company such as
Another thing to consider about disposable gloves.... Depending on the service they will see, it may be important to specify "powder free" gloves. Many use talc, cornstarch, or other lubricant. We had some that seemed to use sodium
hydroxide :-) as much as they irritated my hands! The powder can cover an SEM sample like snow -causing charging and x-ray analysis artifacts.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Message on behalf of Richard Easingwood:
I am trying to compile a list of what type of disposable gloves should be used with what chemicals found in our EM unit. I have compared information from various sources and there seems to be a lack of clear data for some chemicals (although no shortage of opinions from some users). As a result of this survey I have got an idea of what is the general consensus for each chemical, this appears below. I would appreciate any comments, especially if anyone considers me to be definately 'off the mark' on any item:
Some, as you can see, I haven't been able to decide on.I want to limit the gloves to disposables where possible.
Chemical:Glove polymer type (latex, neoprene, nitrile, PVA or polyethylene)
acrylic resinsY' (Lowicryl, LR Gold, Unicryl): Neoprene, nitrile(?)
Thanks in advance to anyone who bothers reading this and responding!
Richard Easingwood
----------------------------------------------------------------------- Richard Lander, NZCS South Campus Electron Microscope Unit c/- Pathology Department Otago Medical School P.O. Box 913 Dunedin New Zealand. Tel. National 03 479 7301 Fax. National 03 479 7254
"Southernmost EM Unit in the World!" ------------------------------------------------------------------------
Message-Id: {2.2.32.19970226141317.006b2c58-at-pop.wwa.com} X-Sender: spb-at-pop.wwa.com X-Mailer: Windows Eudora Pro Version 2.2 (32) Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
If you can help this person, please send a message directly to him at "Richard S. Perla" {RPERLA61-at-mail.caps.maine.edu}
He is not on this listserver.
Dir Sir or Madam,
I am a graduate student in microbiology interested in microscope repair. Do you know of a company of school that has a shot course in repair or a person I may contact to find out further information. Thank you.
Sincerely,
Richard S. Perla 151 West Elm St. Yarmouth, ME 04096
Susanne Pignolet Brandom, Ph.D. MC Services 6-D North Commons Lincoln MA 01773
617-259-0292 617-259-3376
MicroWorld Resources and News http://www.mwrn.com/
Having designed both types of field emission systems in the past, I feel I should add some comments to this discussion. The key to stable field emission is basically good vacuum at the source. The CFEG is particularly sensitive in this respect, but with good UHV design a stability of 2-3% (short-term) and drift of {2%/hr. can be achieved in practice. The Schottky FEG is certainly more stable, but, unlike the Philips report posted by Max Otten, I would not conclude that CFEG is unsuitable for TEM imaging in general. Regarding the energy spread for CFEG, there are two advantages over the Schottky FEG: first, the low temperature of the source results in a smaller energy spread ( {0.3eV); second, the emission current is typically a factor of 10-20 lower than for Schottky sources, so that the energy broadening due to the Boersch effect is less significant, depending on the electron optical design of the illumination system. The Philips data is surprisingly pessimistic (0.7eV at 3nA current), by comparison the VG STEM at 100kV gives typically {0.4 eV energy spread at that probe current. For EELS work requiring high energy resolution the CFEG should be the better choice. Concerning brightness, the CFEG is 5-10 times brighter over a period of several hours between 'tip flashing ' (assuming good UHV conditions). That means more current can be obtained in a given probe diameter, assuming otherwise identical illumination systems, and hence better spatial resolution in EDX and EELS analysis. My conclusion is that both types of FEG have significant advantages and the choice depends on what type of microscopy you want to do. Sebastian von Harrach VG Scientific
Having designed both types of field emission systems in the past, I feel I should add some comments to this discussion. The key to stable field emission is basically good vacuum at the source. The CFEG is particularly sensitive in this respect, but with good UHV design a stability of 2-3% (short-term) and drift of {2%/hr. can be achieved in practice. The Schottky FEG is certainly more stable, but, unlike the Philips report posted by Max Otten, I would not conclude that CFEG is unsuitable for TEM imaging in general. Regarding the energy spread for CFEG, there are two advantages over the Schottky FEG: first, the low temperature of the source results in a smaller energy spread ( {0.3eV); second, the emission current is typically a factor of 10-20 lower than for Schottky sources, so that the energy broadening due to the Boersch effect is less significant, depending on the electron optical design of the illumination system. The Philips data is surprisingly pessimistic (0.7eV at 3nA current), by comparison the VG STEM at 100kV gives typically {0.4 eV energy spread at that probe current. For EELS work requiring high energy resolution the CFEG should be the better choice. Concerning brightness, the CFEG is 5-10 times brighter over a period of several hours between 'tip flashing ' (assuming good UHV conditions). That means more current can be obtained in a given probe diameter, assuming otherwise identical illumination systems, and hence better spatial resolution in EDX and EELS analysis. My conclusion is that both types of FEG have significant advantages and the choice depends on what type of microscopy you want to do. Sebastian von Harrach VG Scientific
I've noted a lot of traffic lately regarding antibodies raised in various animals, secondary antibodies (conjugated and unconjugated), etc. Unfortunately, I have also recently cleaned out my mailbox, so I can't respond to specific persons' questions!
However, I'd like to pass the following information to anyone who works with antibodies/immuno. A great place to visit or to start your search for details on vendors, techniques, monoclonals/polyclonals etc. is the Antibody Resource Page. The URL is:
Whether you're a beginner or an accomplished immuno person, if you have a question this web site either has the answer or can guide you via links to someone who has the answer.
Happy labeling!
Bob Chiovetti
*****The opinions expressed above are my own and are not necessarily shared by any other form of life in the known universe!*****
The CIE Characterization Facility at the University of Minnesota will be sponsoring a Master Class:
"Transmission Electron Microscopy for Materials Science."
Topics
Basic Principles of TEM Imaging and Diffraction Mode Basic Principles of Analytical Elecron Microscopy Basic Concepts of High-Resolution Microscopy Computer Analysis of EELS and EDS Spectra Image Processing and Analysis of Digital TEM Images
May 15-16, 1997
Instructors
John Bruley, PhD, Senior Engineer, IBM. His main research interests involve the study of grain boundaries and interfaces between metals and ceramics, using the analytical microscopy techniques of electron energy loss spectroscopy (EELS) and energy dispersive X-ray analysis (EDX).
Stuart McKernan, PhD, Senior Research Associate, CIE, University of Minnesota. His research involves the study of grain boundaries and interfaces in ceramics.
The Master Class will provide a non-mathematical description of the basic principles of transmission and analytical electron microscopy. The course will be directed towards materials scientists, advanced technicians and technical managers who are required to use or interpret data obtained from the TEM, or who wish to determine whether the TEM is a suitable tool for their requirements.
Lectures in the mornings will introduce the topics and hands-on labs in the afternoons will allow the participants to put their knowledge to use.
Enrollment is limited. Please contact us by return email or phone (612) 626-7594 for additional information (deatailed description, tuition, agenda, etc).
_______________________________________________________________________ Beth Trend btrend-at-tc.umn.edu http://resolution.umn.edu Coordinator, Characterization Facility University of Minnesota Center for Interfacial Engineering 100 Union St SE Room 187 phone 612-624-1365 fax 612-626-7530 Minneapolis, MN 55455
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have found that grids lose some of their "stickiness" for sections within a week of gridwashing. I wash my grids right before I do ultrathin sectioning, and I also add a wash with acetic acid after the acetone and water washes.
Second of all, if you can float your grids on the immunostaining solutions instead of immersing them, your odds will improve. Triton-X or Tween 20 detergents seem quite effective in removing sections from the grids when any grids sink, or are immersed in solutions that contain them.
Gregg Sobocinski Parke-Davis Pharmaceutical Research Division Ann Arbor, Michigan USA Sobocig-at-aa.wl.com
} We have been experienceing a 50% loss of Epon-Araldite embedded thin } sections from nickel grids during immunostaining (Au). We have tried } coating mesh grids with dilute formvar or butvar solutions to no avail. } Can anyone help?
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Subject: Time:8:30 AM } OFFICE MEMO etch Date:2/25/97 } } Does anyone have recipes for "etching" the surface of LR White in order to } expose antigenic sites for surface immunogold locaizations? Also, a recipe or } reference to the procedure for etching Epon or Araldite thin sections for } immunogold. We have used it several years ago but perhaps there is a better } method than NaMethoxide. } thanks } jim jamieson } } Hi, Etching is not thought necessary for LR White thin sections since it is an acrylic with much less propensity to crosslink and bind up antigens than the epoxides. There may even be a reason not to do it. The last issue of J. of Histochem. Cytochem. states that etching may render some antigens less capabable of detection. Should you want a protocol for etching with Naperiodate (preferable to methoxide), let me know by direct e-mail, and I would be happy to supply it.
However, if your wish is to localize ricin on tissue sections and the } conjugation process is not working, then why not try looking for unconjugated, } bound ricin on your samples using anti-ricin antibodies? This may be a much } simpler solution to your problem.
With the extreme toxicity of Ricin, is it even possible to make bunny bodies?
I never recommend methods I haven't tried before. If I do, I make sure it is clearly a speculation. So I guess it must be possible to make anti-ricin antibodies, I have used them before.
Regards,
Paul Webster Center for Cell Imaging Yale School of Medicine http://info.med.yale.edu/cellimg
Jeol Australia tells us that mercury reference batteries are no longer available so we must design and manufacture power supplies for the high voltage and focussing reference voltages.
Has anyone been down this track already? Are there circuit diagrams available?
Any help would be greatly appreciated.
Eric Hines Microscopy Centre CSIRO Entomology Canberra Australia
I have followed the discussion about cold and thermal FEG=B4s at high KV in TEM units. But are the advantages or disadvantages of thermal Field emitters at low KV in a SEM.
Thanks
Heike Buecking Dr. Heike Buecking Universit=E4t Bremen, UFT Physiologische Pflanzenanatomie Leobener Str. 28359 Bremen Tel: +49-0421-218-2954 -7283 Fax: +49-0421-218-3737
Years (or decades) ago we had to do something similar for a Siemens IA although we were lucky because Siemens had their own circuit and installed it for us.
The system worked fine, but of course it was not as sensitive a beast as a JEOL 100.
Malcolm Haswell University of Sunderland UK ----------
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America -----------------------------------------------------------------------.
Dear all,
Jeol Australia tells us that mercury reference batteries are no longer available so we must design and manufacture power supplies for the high voltage and focussing reference voltages.
Has anyone been down this track already? Are there circuit diagrams available?
Any help would be greatly appreciated.
Eric Hines Microscopy Centre CSIRO Entomology Canberra Australia
A friend of a friend is looking for a morphometric system with Scope, stylus, PC etc. They saw some at the recent Neurosci meetings, but didn't keep the brochures. Any info would be greatly appreciated.
Thanks
Cheri Owen Detroit Neurotrauma Institute Wayne State University Detroit, MI
We have acquired a limited quantity of factory-new Mitsubishi CP-10U color video printers; if you are interested, the closeout price is under $500.00/ea.
Contact:
Shawn Oliver Vision Quest, Inc. 1-800-284-4140 ext. 304, or 1-417-862-1967 ext. 304 www.visionquest-cctv.com
A while back I read a paper in which a test specimen was prepared for nearfield scanning microscopy by spin coating beads mixed in polyvinyl alcohol onto a coverslip. They gave times and RPM's, and quoted a resulting sample thickness of tens of nanometers.
Does this mean the coverslip is placed on the bottom of a swinging bucket centrafuge rotor, sample placed on the coverslip, and spin?
Or is the coverslip in some other orientation? Is there a special "spin coating" apparatus?
The many requests for information and protocols on etching of thin sections prior to post-embedding colloidal gold procedures has prompted me to try to answer all of you at once.
1. Etching as a preliminary treatment of thin sections destined for pot-embedding colloidal Au is indicated for expxide sections ONLY. Epoxy resins react with tissue components, acrylics (LR White) do not, but crosslink through the tissue and not with it (Newman, Resin Microscopy and On-Section Innumocytochemistry, Springer Verlag, pa. 132). Thus, etching of acrylics may loosen the tissue from its support. Further, acrylics are far more hydrophylic than epoxides.
2. Strong oxidizing agents may temporarily render the surface of the normally hydrophobic epoxides hydrophylic, and they remove osmium bonds thus increasing the chances for labelling of exposed antigens. Bendayan and Zollinger (J. Histochem. Cytochem. 31:101, 1983) have shown that the optimum agent for increasing labelling of antigens on the surface of expoxide sections is saturated sodium periodate. It unmasks antigens by removing osmium. Please note: Contrast will be reduced.
3. In our laboratory we have found that sodium m-periodate is superior to methoxide. Methoxide is too difficult to standardize, is likely to punch holes into the sections, and otherwise be detrimental to the process causing section instability in the TEM. We use sodium m-periodate (Sigma, Cat# S-1878).
4. Every embedding formulation requires different "etching" time. We use a very soft (low crosslinkage) formulation using Araldite-Epon-DDSA, which is relatively easily swelled by water based liquids. Our gold thin sections require only 5 minutes in periodate. Other formulations may require up to 15 minutes or more. To detect the optimum times for a specific embedding medium, trial thin sections should be exposed to saturated periodate for 2,5,10,15,20 min, and then examined by TEM before a large number of grids are committed to the procedure.
A. Make a saturated solution of sodium m-periodate using 1g of perodiate in 5 ml of ddH2O. B. Float section (on nickel grids) on 50microliter drops of periodate C. Rinse WELL. Pass grids through 6 changes of ddH2O 2 min each.
CONSIDER NOT ETCHING EPOXY SECTIONS AT ALL: Use the method of Phend, which totally eliminates osmium in the fixation process. We like this the best, and have gotten an increase in labelling of our sparse antigen with this procedure. (Phend et al., J.Histochem. Cytochem., 43:283-292, 1995)
Our Kevex 8000 EDS system has horrible 8 inch 10MByte Bernoulli drives. For the first time in a year or so we need to buy some new disks. Does anyone know where to buy them? Our local vendors won't carry them anymore. Kevex (in typical fashion) is being completely uncooperative.
By the way, sometime in the future we will need to purchase a new EDS system. We will never buy another Kevex system (see above). Does anyone have any opinions about what system gives both the best performance and the best service?
} A while back I read a paper in which a test specimen was prepared } for nearfield scanning microscopy by spin coating beads mixed in } polyvinyl alcohol onto a coverslip. They gave times and RPM's, } and quoted a resulting sample thickness of tens of nanometers. } } Does this mean the coverslip is placed on the bottom of a swinging } bucket centrafuge rotor, sample placed on the coverslip, and spin? } } Or is the coverslip in some other orientation? Is there a special } "spin coating" apparatus? } }
Spin coating is very common in microlithography, all photoresists are put onto wafers in this way, so there is a "spin coating apparatus." It consists of a flat chuck onto which you would place the slide flat. A vacuum holds the slide down onto the chuck. You pour some liquid onto it and spin at at as constant an RPM as possible for about 30 seconds. 3000-5000 RPM is normal. The more viscous the fluid the thinner the resulting layer. This is probably the procedure they used.
Spin coaters are common in the semiconductor industry. Basically, they are a high speed motor with a vacuum chuck attachment on the top of the shaft. The substrate is held flat to the top of the shaft by the vacuum. Doped PVA films may be spun to thicknesses of ~} 10 nm easily. Depending upon the viscosity of the media you might try values of 5,000 RPM for ~ 30 seconds.
-------------------- At 11:42 AM 2/27/97 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Edward Spin coating is more like setting a cover slip on the center of a record player and applying a liquid while it is rotating. We use a spin coater manufactured by Headway Research from Garland, Texas.
cheers
Ed Basgall Penn State Univ. Dept. of Chemistry University Park, PA
----- Begin Included Message -----
From Microscopy-request-at-Sparc5.Microscopy.Com Thu Feb 27 16:10:27 1997
On Thu, 27 Feb 1997, Cheri Owen wrote:
} A friend of a friend is looking for a morphometric system with Scope, } stylus, PC etc. They saw some at the recent Neurosci meetings, but didn't } keep the brochures. Any info would be greatly appreciated.
A more detailed explanation of the intended tasks would help.
I need to buy a vibratome and it has been a while since I used one. Does anyone have a brand they are happy with and where it was purchased.
Thanks in advance.
Ruth Yamawaki ******************************************************* Ruth Yamawaki Department of Comparative Medicine Stanford University (415) 723-3457 ***************************************************************
Does anyone want , for freight costs only , a complete , but currently not working Quantimet 900 Image analysis system ? The dedicated Newvicon video camera is inoperative and will have to be repaired / replaced.
If interested please contact me directly : griffiths.michael.mj-at-bhp.com.au
Thanks from B.H.P. Steel , Newcastle , New South Wales , Australia
I would appreciate it if anyone who has a specification for the EMSA ASCII eds spectrum file format could send me a copy. ************************************************* * Bill Hardy, President * * American Nuclear Systems, Inc. * * 12633 Red canyon Road * * Knoxville, TN 37922 * * (423) 671-0292 FAX: (423) 671-0293 * * WWW.qtmsys.com Email: bhardy-at-qtmsys.com * *************************************************
We published a short technical note on alternative power supplies for the JEOL 100CX in our South African EM Conference Proceedings back in 1988. Although I believe that more recent developments in electronics may improve on this design, the simple unit described in our paper has worked just fine for us since 1988 and still going strong. If lots of folk would like the design I could scan it, otherwise just send me your fax number.
Tony Bruton Head, Centre for Electron Microscopy University of Natal, Pietermaritzburg Private Bag X01, Scottsville. 3209 KwaZulu-Natal, South Africa Tel +27 331 2605155 Fax +27 331 2605776 Email: bruton-at-emu.unp.ac.za
} } } HASWELL Malcolm {es0mhs-at-environment.sunderland.ac.uk} 27/February/1997 02:37pm } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com -----------------------------------------------------------------------.
Years (or decades) ago we had to do something similar for a Siemens IA although we were lucky because Siemens had their own circuit and installed it for us.
The system worked fine, but of course it was not as sensitive a beast as a JEOL 100.
Malcolm Haswell University of Sunderland UK ----------
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America -----------------------------------------------------------------------.
Dear all,
Jeol Australia tells us that mercury reference batteries are no longer available so we must design and manufacture power supplies for the high voltage and focussing reference voltages.
Has anyone been down this track already? Are there circuit diagrams available?
Any help would be greatly appreciated.
Eric Hines Microscopy Centre CSIRO Entomology Canberra Australia
} I need to buy a vibratome and it has been a while since I used one. Does } anyone have a brand they are happy with and where it was purchased.
"Vibratome" is the registered trademark of Technical Products International, and the name refers to a specific instrument (once made by Oxford Instruments, then Sherwood Medical, and now, TPI). There are three authorized United States distributors for the instrument: Energy Beam Sciences, Ted Pella, Polysciences, Kramer Scientific and Baxter (or whatever they're calling themselves these days). (Technical Products International does not sell the instrument direct, only through distibutors.)
Several companies manufacture *other* vibrating blade microtomes (please, not "vibratomes"), including Energy Beam Sciences (the "MicroCut"), Dosaka (the "MicroSlicer"), Camden Instruments (the "Vibroslice"), Leica, and EMS.
I assume my biases are obvious {grin} .
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
I have a couple of questions concerning LR White, and know that this topic was discussed in some length a while back.
We would like to use some traditional botanical stains to stain non-osmicated LR White sections of plant material. These light microscopy stains that we would like to use, are designed for use with deparaffinized sections.
The questions are:
1) Can these stains be used directly on the LR White sections, or do they have to be modified in some way?
2) Does the LR White have to be removed before the stains are applied?
I know that people were having some problems with their sections floating away when they applied some stains, and Thomas Phillips posted a solution to this problem by using aminopropy-triethoxysilane. Has anyone else tried this technique or have some other good suggestions?
Please respond to me directly if this topic is redundant.
Thanks in advance,
Susan
Susan Carbyn Atlantic Food and Horticulture Research Station Kentville, Nova Scotia B4N 1J5 Canada
Thanks for all of the responses. The correct reference to the two versions of the EMSA file format can be found at:
The first reference is to an FTP directory where the original specifications are located. The DOC files contain the info. For PC users note the file format is not standard PC but can be read in Microsoft Word. You will need this because the second is not a complete specification.
ftp://ftp.msa.microscopy.com/pub/4-MMSLib/XEDS/EMMFF/ for version 1.0
The second directory has additional information.
ftp://ftp.msa.microscopy.com/pub/4-MMSLib/MISC/RWEMMPDL/ for version 1.1
Once again thanks everyone.
Bill Hardy ************************************************* * Bill Hardy, President * * American Nuclear Systems, Inc. * * 12633 Red canyon Road * * Knoxville, TN 37922 * * (423) 671-0292 FAX: (423) 671-0293 * * WWW.qtmsys.com Email: bhardy-at-qtmsys.com * *************************************************
We recently moved our TEM and SEM from a building built in 1930 to one built in 1995. In other words, to a building with sprinklers. Now each microscope has a sprinkler directly overhead and I've been wondering if I should cover the scopes at night -- just in case. Do other people have this concern/problem? I haven't read anything about sprinklers malfunctioning and going off but it is a bit unnerving seeing a shower head directly over the scopes! I haven't seen covers in any of the catalogs so assume a tarp is the way to go....Or is it?
Thanks for any feedback....
Peggy
Peggy Brannigan Electron Microscopy Floral and Nursery Plants Research Unit National Arboretum
} I am trying to compile a list of what type of disposable gloves should be } used with what chemicals found in our EM unit. [snip] } } Organic solvents: } } acetone: butyl? or latex } ethanol: latex } methanol: latex } chloroform: (PVA if avail) or nitrile (double glove) } [snip]
Dear Richard, I'd be very surprized if latex gloves would hold up to acetone; when I've exposed them, they get tacky. Polyethylene gloves should hold up to the organic solvents you listed (and many others). Besides, they smell better than latex gloves. Yours, Bill Tivol
In message {v03007803af3c74e4972a-at-[198.77.169.26]} Peggy Brannigan writes:
} Now each } microscope has a sprinkler directly overhead and I've been wondering if I } should cover the scopes at night -- just in case. Do other people have } this concern/problem? I haven't read anything about sprinklers } malfunctioning and going off but it is a bit unnerving seeing a shower head } directly over the scopes! I haven't seen covers in any of the catalogs } so assume a tarp is the way to go....Or is it? } } Thanks for any feedback.... } } Peggy
I strongly advise you to put some kind of water deflector over your scopes. We put up plexiglass "roofs" over our TEM and SEM because of occasional water dripping through cracks in the concrete ceiling due to mishaps in floors above us. The roofs are usidedown V-shaped, /\ , so shed any water off to the sides of the scopes to the floor. They have saved us a few times since they were installed. They also prevent some dust from accumulating on the scopes. They are pivoted at the vertex and adjustable to allow for changing light bulbs. Our carpenter shop at physical plant put them up.
In your case, better check with the building saftey inspectors vis a vis the fire issue.
Gib
--
Gib Ahlstrand, MMS Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
} We recently moved our TEM and SEM from a building built in 1930 to one } built in 1995. In other words, to a building with sprinklers. Now each } microscope has a sprinkler directly overhead and I've been wondering if I } should cover the scopes at night -- just in case. Do other people have } this concern/problem? I haven't read anything about sprinklers } malfunctioning and going off but it is a bit unnerving seeing a shower head } directly over the scopes! I haven't seen covers in any of the catalogs } so assume a tarp is the way to go....Or is it? } Dear Peggy, If the scopes have diffusion pumps, or any other heat source, which might start a fire, do *not* cover them in such a way as to block ventil- lation. Maybe you could get the sprinklers replaced by halon units or some other extinguishing system better suited for electrical fires. A fire would likely involve electrical systems even if it was not started that way, and water is not the right extinguisher for this kind of fire. If the law says that sprinklers are necessary even if they will do only harm, then you could be stuck. This would be another place where the "obvious stupidity exception" would be a good thing for the law. Good luck. Yours, Bill Tivol
Sprinklers can and do go wrong. We are on the second floor and the sprinklers went off on the sixth floor. The water made it all the way down here. Luckily we had enough warning to scrounge up enough plastic to cover down the scopes, computers etc. Folks on the fifth floor were not so lucky. Their ceilings collapsed and they we ankle deep in water. By the way some laser printers will float.
My microscope service man suggests buying matress bags from a moving company. They will slip right over the column.
An additional worry might be the iron pipes from the sprinklers giving you field problems with your scope. All depends on how sensitive your instrument is. When we had sprinklers installed we had them placed on the wall about 4 ft. from the column, just for that reason . In another room where that was not possible we had them use plastic pipe for the part that crossed over the column. } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } At 11:40 AM 2/28/97 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
In a continued effort to support all of our 8000/Delta users, Kevex has just released a NEW disk drive upgrade kit. This will replace the old Bernoulli drive system that many customers are still using. The new drives will replace the present drive system and still use the same interface board, thus saving the customer that added expense. The problem with the Bernoulli drive systems (both the 10Meg and 44Meg systems) is that the media and drives are no longer available. Iomega Corporation will no longer make or support these drives or media. This is beyond Kevex Instruments control.
Kevex will always do EVERYTHING possible to support older instruments that we manufactured for as long as possible. Kevex still supports the 7000 systems that where manufactured 19 years ago. In this day of computer technology, this is no easy feat.
Thanks for all the responses on where to get 8 inch, 10 MByte Bernoulli disks. I also got an immediate response from the Kevex rep. when he read my request on the list server. We must have just gotten hold of the wrong people at Kevex.
I would be careful about covering the scopes with a tarp on a routine basis since this could generate other problems. For example, heat from the consoles could cause damage to the electronics or if somebody pulls on the tarp they could inadvertently damage a holder or an aperture control. I would first consider moving the sprinkler. Check with your safety department.
By the way, we do not have a sprinkler in our lab, but a few years ago there was one night a water leak in the lab above ours and in the morning we had a nice cascade coming down (it barely missed the scope). On another occasion the same lab (above ours) had a fire and we were again flooded , this time by the fire department. In both instances power to the lab was turned off and the scope survived just fine ! I am worrying about the third strike !!!.
One of the faculty members in my department is looking for information on software for image analysis of animal chromosomes. The program she was most interested in is being discontinued. She would appreciate any advice on "reasonably priced" (less than $10K) software for FISH and Karyotyping, recommended models of ccd cameras, and filter wheels to fit her Zeiss Universal. Ideal situation would be a vendor capable of supplying all components, installing them and supplying software training. Information from persons doing this type of work would be especially appreciated.
Please send information to Ruth Phillips, rp-at-csd.uwm.edu.
Thank you,
Heather Owen
Heather A. Owen, Director Electron Microscope Laboratory Department of Biological Sciences University of Wisconsin - Milwaukee (414)229-6816
NOTE: please reply directly to me at wrightr-at-zoology.washingtonl.edu
I am interested in purchasing a new diamond knife and was really suprised at the great prices available from Micro Star. If you have used a Micro Star knife for ultramicrotomy, I would appreciate hearing PRIVATELY about your experience.
Robin Wright University of Washington Department of Zoology, Box 351800 Seattle, WA 98195 Phone: 206-685-3651 FAX: 206-543-3041
Gentlemen: I publish a medical newsletter authored by Thomas A. Dorman, M.D. Our readership can best be described as "educated laymen" (although there are a number of physicians who subscribe.)
A recent flyer I received in the mail stated that some 12 blood disorders can be identified, including free-radical damage, liver stress, heavy metals, etc. I could not tell how accurate the information was; in fact, it invited the public to come for a free demonstration, which was followed by sell-job (misc. pills) What struck me was that those who came had not been told to fast, which if I remember right is suggested. (Maybe I'm wrong on this?)
Long story short, we would like to publish an authoritative article that provides the interested reader a clear overview of what Darkfield microscopy is, and what practical value it has for him/her. Would you be interested in supplying an article dealing with this fascinating subject, along with photos of actual blood cells and the various blood disorders that can be identified?
Thank you for your kind attention to this request.
My EM unit is located in an old building therefore we do not have the "sprinkle system". However, the water may leak out from the very old pipes, so we hang up a 4 x 5 meters canopy made of woodframe and poly sheet right above the EM. So far we never worry about any downpour of water on our scopes.
Regards,
On Fri, 28 Feb 1997, Peggy Brannigan wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } We recently moved our TEM and SEM from a building built in 1930 to one } built in 1995. In other words, to a building with sprinklers. Now each } microscope has a sprinkler directly overhead and I've been wondering if I } should cover the scopes at night -- just in case. Do other people have } this concern/problem? I haven't read anything about sprinklers } malfunctioning and going off but it is a bit unnerving seeing a shower head } directly over the scopes! I haven't seen covers in any of the catalogs } so assume a tarp is the way to go....Or is it? } } Thanks for any feedback.... } } Peggy } } Peggy Brannigan } Electron Microscopy } Floral and Nursery Plants Research Unit } National Arboretum } } Bldg. 010A R.238 } 10300 Baltimore Avenue } Beltsville, MD USA20705 } } Phone: (301) 504-6097 } Fax : (301) 504-5096 } Email: brannign-at-asrr.arsusda. gov } } } }
*********************************************** * Ming H. Chen, PhD * * Medicine/Dentistry Electron Microscopy Unit * * University Of Alberta. * * Edmonton, Alberta, Canada * * * * Visit My Page At: * * http://www.ualberta.ca/~mingchen * ***********************************************
The software for a conversion of AN10000 to Windows has already been written by a Toronto based company called Mektech. They also make a hardware to interface the AN10000 pulse processor to a PC. You can find them at mektech-at-visionol.net The Microscopy Imaging Laboratory at the University of Toronto Dept. of Medicine. Has purchased the interface for the AN10000 to PC and the software from Mektech. The software is easy to operate and we have found it to be very reliable. Mektech has allowed us to upgrade our system at a fraction of the cost of that of a new system.
Battista Calvieri Manager of Microscopy Imaging Labarotory University of Toronto
Not even the suggested sophistication of a "halon" fire protection system will render your EMs immune from the dreaded "water shower" experience. Overnight flooding of the lab floor above us caused water to find its way to (and then through) the smoke/ thermal detectors that we had smugly positioned directly over the instrument for our halon gas quench system, cascading water onto a powered TEM/STEM/EDS facility. By the time I was called in, the TEM was lit up like a Xmas tree, accompanied by the delicate aroma only smoldering electronics can generate.
Unfortunately, similar water showers in the adjacent SEM room had missed the console, and the instrument was too heavy to push under the water stream (we were trying to get a new SEM at the time!).
Note: Our fire detectors have since been relocated, and the "halon" has been replaced (by Govt decree) with environmentally more friendly (less unfriendly?) FM200.
Eric Bradley BHP Steel Port Kembla NSW Australia bradley.eric.eg-at-bhp.com.au
---------------------------------------
Some months ago a gate valve went in the cooling pipes for our building, which created a spectacular waterfall in the corridor. Since an insurance claim is the easiest way of getting new equipment here, we were quite hopeful, but like Eric Bradley, we found that three strong people were unable to push either our 17 year old TEM, 25 year old Auger kit, or even the little 18 year old SEM under the water.
I have a problem about cell number count on agrregated neurons with an enzyme treatment. I want to check a viability after the enzyme treatment. It is usually available to count a cell number under phase contrast microscopy, but light was so reflected that each neuron could not be identified. I would, therfore, try to apply a fluorescence dye,for example, Hoechst33342, that bind to a minor grove of DNA and is heared to be applicable to a native cell. But I wonder it will work well. I would like to get more detailed information about image analyze of cell number under a microscopy. Thanks a lot.
Masahito Morita Laboratory of Compararive physiology Faculty of Science Osaka University
} I am trying to compile a list of what type of disposable gloves should be } used with what chemicals found in our EM unit. [snip] } } Organic solvents: } } acetone: butyl? or latex } ethanol: latex } methanol: latex } chloroform: (PVA if avail) or nitrile (double glove) } [snip]
Dear Richard, I'd be very surprized if latex gloves would hold up to acetone; when I've exposed them, they get tacky. Polyethylene gloves should hold up to the organic solvents you listed (and many others). Besides, they smell better than latex gloves. Yours, Bill Tivol
Richard, and others interested;
We use 100% Nitrile gloves as they stand up to all the chemicals we use in the EM lab, We use the product available from Best Manufacturing Company, Menlo, GA 800-241-0323. I'm sure there are others just as suitable.
Disclaimer: We have no financial or other interest in the company.
But I've been on the road for awhile and hence did not have time to catch up on things. Here's the info that a few people have asked for as I did not see anyone else post an answer.
The MSA/MAS Spectrum File Format Information & Examples can be downloaded from the following FTP Site.
Host: ftp.amc.anl.gov UserID: anonymous Password: Your Email Address
Everything is in the EMMFF ( "E"MSA MAS File Format) directory, including examples and documentation.
Version 1.0 is the current revision level. We (The MSA/MAS Standards Committee) have not had a need to update it beyond that point.
I have not gotten around to creating a WWW page for this. If there is enough interest I will do it. However if you have access to the WWW you can get it via FTP. It is also routinely available in the Computer Workshop at the annual Microscopy & Microanalysis Meeting . ( see http://www.msa.microscopy.com for this years meeting information )
Cheers... Nestor Your Friendly Neighborhood SysOp
Here is the Abstract File for that directory.
-------- Title :EMMFF V. 1.0 Keywords :XEDS,EELS,AES,WDS,CLS,GAM,XRF,PES Computer :IBM, MAC, DEC Operating System :ALL Programming Language :Fortran 77 Hardware Requirements :None Author(s) :EMSA/MAS TASK FORCE Ray Egerton ,Charles E. Fiori ,John A. Hunt, Mike S. Isaacson,Earl J. Kirkland ,Nestor J. Zaluzec Correspondence Address :R.F. EGERTON-CHAIRMAN University of Alberta Dept. of Physics Edmonton, Alberta, Canada, T6G2J1 Abstract:
A simple format for the exchange of digital spectral data is presented, and proposed as an EMSA/MAS standard. This format is readable by both humans and computers and is suitable for transmission through various electronic networks (BITNET, ARPANET), the phone system (with modems) or on physical computer storage devices (such as floppy disks). The format is not tied to any one computer, programming language or computer operating system. The adoption of a standard format would enable different laboratories to freely exchange spectral data, and would help to standarize data analysis software. If equipment manufacturers were to support a common format, the microscopy and microanalysis community would avoid duplicated effort in writing data-analysis software. This version of EMSAMASFF contains two subroutines which read and write spectral data files Version 1.0 data format. The data are stored as simple ASCII characters at a user defined number of columns per line for the length of the data file. The spectral data is preceeded by a series of header lines, which tell the user about the parameters of the spectrum. The header lines are identified by the first character in the line being the symbol (#) followed by a descriptor and if appropriate its units. An example of a data file format can be found in the EMSAMASFF.DOC file. ------------------------------------------------------------------------------
You never know what you will find in your e-mail "in" basket. If any of you know the derivation of the word "Wehnelt", please respond to this gentleman directly at:
} The subscriber is obviously a relative of this "wehnelt" you mention. I } belong to the Brazilian side of the family, hence the different graphy of } the name. Can you be kind enough to give more info on this man? I knoe there } was an Arthur Waehneldt who dedicated himself to physics. Are we } talking about the same person? Pls. send all the details you may have. I`m } trying to trace member oof the family ald your cooperation will be very } wellcome. Thanking you in advance, I remain. } Gustavo } gustavo waehneldt } guswae-at-tricom.net ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
Good morning, I would like to thank everyone who so helpfully provided information on servicing the stereo pod on my Ultracut:
Lou Ann Miller, Julian Smith III, Normand Laurier, Allan Mitchell, Kevin Hlacrow, Sara Miller, Dan Focht, Geoff McAuliffe, and Alexander Greene.
I appreciate, as always, having access to such a generous group of people, who find the time to provide useful information. Thanks again!
Dwight Beebe E-mail: beebed-at-ere.umontreal.ca Institut de recherche en biologie vegetale Voice: 514-872-4563 Universite de Montreal FAX: 514-872-9406 4101, rue Sherbrooke est Montreal, Quebec H1X 2B2 Canada
So it takes more than "three strong people" to push them under? I'll bear that in mind!
We are still running an Edwards 12E6 coating unit coming up to 32 years old (can anybody beat that?), it goes like a Trojan (great for ploughing, farming joke!).
Regards - Keith Ryan Plymouth Marine Laboratory, UK
Perhaps I state the obvious, but beware of (powder) lubricated gloves. When I started SEM work (long, long ago), only powdered gloves were used for specimen prep. - what a mess! It seemed to always be "snowing" on samples sent to me from our failure analysis lab! Although somewhat difficult to locate at the time, I managed to convince all to go with powder-free. No more problem. Powder free are now easy to find.....
I know you have received alot of replies about this subject, but I can't resist adding my experiences. I compare having a water sprinkler system in an EM lab to gambling in Vegas; sometimes you win, sometimes you lose. We have a sprinkler system directly over our Cambridge 200 SEM, and fortunately we have not had to use it in 10 years - doesn't mean it won't happen tomorrow, though. We DID, however, have our ceiling cave in on our scope during a heavy rain. The soggy composite tiles were like chewed up spitwads all over the unit, and a steady river of water was running from the roof directly into the keyboard and console. Figuring that it was probably dead, I called the manufacturer for advise. We fixed the leak, then opened the SEM up and dried everything the best we could with blasts from a nitrogen tank. We allowed it to dry for 3 more days and fired it up. No problems other than some sticky keys in the keyboard - pretty amazing.
(BTW, this is not an advertisement for Cambridge/LEO. It's a real experience)
Regards,
-Bob ******************************** Bob Citron Chiron Vision 555 W. Arrow Hwy Claremont, CA 91711 ph: (909)399-1311 email: Bob_Citron-at-cc.chiron.com ******************************** ______________________________ Reply Separator _________________________________
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We recently moved our TEM and SEM from a building built in 1930 to one built in 1995. In other words, to a building with sprinklers. Now each microscope has a sprinkler directly overhead and I've been wondering if I should cover the scopes at night -- just in case. Do other people have this concern/problem? I haven't read anything about sprinklers malfunctioning and going off but it is a bit unnerving seeing a shower head directly over the scopes! I haven't seen covers in any of the catalogs so assume a tarp is the way to go....Or is it?
Thanks for any feedback....
Peggy
Peggy Brannigan Electron Microscopy Floral and Nursery Plants Research Unit National Arboretum
This world wide E-mail provides important information for everyone involved in biomedical research. We hope no one feels bothered by this mail. Spamming is not indented.
Dear researcher,
Carl Zeiss is very pleased to announce it's new high performance confocal
Laser Scanning Microscope LSM 510,
designed for your best results in biomedical research.
(L)et's (S)ee (M)ore for applications, technical data and world wide expert contacts
at http://www.zeiss.de/mi/limi_e/p4/lsm510_entry.htm
This world wide E-mail provides important information for everyone involved in biomedical research. We hope no one feels bothered by this mail. Spamming is not indented.
Dear researcher,
Carl Zeiss is very pleased to announce it's new high performance confocal
Laser Scanning Microscope LSM 510,
designed for your best results in biomedical research.
(L)et's (S)ee (M)ore for applications, technical data and world wide expert contacts
at http://www.zeiss.de/mi/limi_e/p4/lsm510_entry.htm
As an anthropological electron probe microanalytical (EPMA) project we are analyzing ancient and diagenetic bone for strontium, barium and other trace elements. We are seeing an unexpected (so what else is new) correlation of Sr with phosphate and the i nverse with apatite replaced by carbonate. We are trying to eliminate analytical errors, and would like to throw this problem at the EPMA community for feedback ... my standard apatite with no Sr measures at detection, and we have double-checked for backg round interferences, but only for elements suspected. We are suspicious of not analyzing for a element which might be causing this problem ... TIA
cheerios, shaf
{\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/epmahome/
Does anyone know of a good preferential etch for P+ silicon? I've been using a 20:1 HNO3:HF stain. This stain seems to delineate the N+ areas very well, but the P+ areas are not easily seen.
I would like to decorate the P+ with as little attack to the N+ areas as possible.
Any help is greatly appreciated.
Regards, Bob Heiman ------------------------------------- E-mail: heiman-at-gmtme.com Voice: 610-666-7950 x2855, FA Lab x2533
Message-Id: {199703031915.OAA14276-at-post-ofc04.srv.cis.pitt.edu} To: microscopy {microscopy-at-Sparc5.Microscopy.Com}
-- [ From: Simon Watkins * EMC.Ver #2.5.02 ] --
As another tale of woe to follow up on Bob Citrons comment on soggy ceiling tiles forming spit wads all over his SEM.
A few years back while I was working in Boston we tried to drown a 100CX by backing up several gallons of water down an air handling chute directly on top of the column (don't ask why or how, but it was one of the side effects of being in the basement). Being on the ball, the operator recognised the problem due to the high moisture content in the air and the splishy splashy sound of water falling from the ceiling. He hit the magic red button to shut the instrument down, and ran for help shouting as he ran "the microscopes drowning, the micrscopes drowning".
As was the case with Bob we called the manufacturer, in this case the ever helpful folks at JEOL, who agreed this was something of a problem but had no immediate solution beyond "errrr, huh, interesting", so we dried off what we could and left the thing off for a week. We turned it on and away it went, as if nothing had happened. The only long term effect was some rusting of the Allen bolts used to hold the column together, you would think that they would be painted or stainless with all these "environmental" problems of late!
my 2 cents
Simon
-- ------------------------------------------------- Simon C. Watkins Ph.D Associate Professor Director Center for Biologic Imaging University of Pittsburgh Pittsburgh PA 15261 tel 412-648-3051 fax 412-648-2004 -----------------------------------------------
The book, Handbook of Metal Etchants, has about a hundred pages of etchant recipes for Si wafers, crystals. Good discussions on practices/ applications as well. Includes references.
Handbook of Metal Etchants Walker, Perrin & Tarn, William H. 1991 CRC Press, Boca Raton, FL 33531 ISBN 0-8493-3623-6
Happy mixing! ------------------------------------------------ Opinions or statements expressed herein, rational or otherwise, do not necessarily reflect those of my employer.
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 Phone: (508) 750-1717 E-mail: crossman-at-osi.sylvania.com
Our web sites: www.sylvania.com www.siemens.com --
Thanks to all who answered my recent question about covering EM's against sprinkler damage! A lot of good points were made, including:
1. The threat is real (some real horror stories out there!) 2. It's not just sprinklers that threaten, it's floods from burst plumbing, storms etc... 3. Tarps are inconvenient and possibly hazardous, 4. Curved plexiglass or fiberglass structures attached to the ceilings over the microscopes were widely recommended (like salad bars) 5. Many suggested dismantling or plugging the sprinkler heads while 6. others were concerned about fire safety 7. A surprising number of drenched scopes fully recovered 8 which you should keep in mind if you actually push your scope under the downpour in hopes of obtaining a new one. 9. but if you try anyway, it takes more than three people.
Thanks again!
Peggy
Peggy Brannigan Electron Microscopy Floral and Nursery Plants Research Unit National Arboretum
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Subject: Time:1:27 PM OFFICE MEMO Why am I not surprised? Date:3/3/97
Last week I received a message from Microscopy about "Little Jessica Mydeck". A check at http://www.cancer.org/acs.html revealed:
Fraudulent Chain Letter (This statement may be copied or reprinted by online users)
The American Cancer Society is greatly disturbed by reports of a fraudulent chain letter circulating on the internet which lists the American Cancer Society as a "corporate sponsor" but which has in no way been endorsed by the American Cancer Society.
This letter appears to have started on America Online but has now spread well beyond the online service. There are several variations of this letter in circulation. The text of the original message reads as follows:
LITTLE JESSICA MYDEK IS SEVEN YEARS OLD AND IS SUFFERING FROM AN ACUTE AND VERY RARE CASE OF CEREBRAL CARCINOMA. THIS CONDITION CAUSES SEVERE MALIGNANT BRAIN TUMORS AND IS A TERMINAL ILLNESS. THE DOCTORS HAVE GIVEN HER SIX MONTHS TO LIVE.
AS PART OF HER DYING WISH, SHE WANTED TO START A CHAIN LETTER TO INFORM PEOPLE OF THIS CONDITION AND TO SEND PEOPLE THE MESSAGE TO LIVE LIFE TO THE FULLEST AND ENJOY EVERY MOMENT, A CHANCE THAT SHE WILL NEVER HAVE. FURTHERMORE, THE AMERICAN CANCER SOCIETY AND SEVERAL CORPORATE SPONSORS HAVE AGREED TO DONATE THREE CENTS TOWARD CONTINUING CANCER RESEARCH FOR EVERY NEW PERSON THAT GETS FORWARDED THIS MESSAGE. PLEASE GIVE JESSICA AND ALL CANCER VICTIMS A CHANCE.
IF THERE ARE ANY QUESTIONS, SEND THEM TO THE AMERICAN CANCER SOCIETY AT ACS-at-AOL.COM
As far as the American Cancer Society can determine, the story of Jessica Mydek is completely unsubstantiated. No fundraising efforts are being made by the American Cancer Society in her name or by the use of chain letters. Furthermore, the email address ACS-at-AOL.COM is inactive. Any messages to the American Cancer Society should be instead sent through the American Cancer Society website at http://www.cancer.org.
I would like to get an explanation for the following observation of quantitative analysis by EDS.
I am looking at a Si wafer with 100nm of SiO2 layer on top. from a SEM. Electron beam (~8 kev) hit the surface along the surface normal (0 deg tilt). As I increase the sample tilt (upto 45 deg or so) EDS quantitative analysis shows a decrease of O2%. How this is possible? I expected to see O2% goes up with the tilt as electron will see more of the surface than bulk.
Could someone explain why?
Thanks
Fran.
--------------------------------------------------------- Get Your *Web-Based* Free Email at http://www.hotmail.com ---------------------------------------------------------
Many thanks for the many recent replies to my earlier queries on Disposable gloves and multi user security. We now have a few more ideas on where to go regarding security in our Unit. The gloves issue is very complex too, mainly due the wide variety of chemicals and conditions they are used for in the lab!
Thanks again,
Rich.
----------------------------------------------------------------------- Richard Lander, NZCS South Campus Electron Microscope Unit c/- Pathology Department Otago Medical School P.O. Box 913 Dunedin New Zealand. Tel. National 03 479 7301 Fax. National 03 479 7254
"Southernmost EM Unit in the World!" ------------------------------------------------------------------------
fransisco black wrote: } Dear Microscopists: } I would like to get an explanation for the following observation } of quantitative analysis by EDS. } } I am looking at a Si wafer with 100nm of SiO2 layer on top. } from a SEM. Electron beam (~8 kev) hit the surface along the } surface normal (0 deg tilt). As I increase the sample tilt } (upto 45 deg or so) EDS quantitative analysis shows a decrease } of O2%. How this is possible? I expected to see O2% goes up } with the tilt as electron will see more of the surface than } bulk. } Could someone explain why? } Thanks } Fran.
Fran, My initial guess would be the change in the sample-detector geometry. Although one would expect the O2 counts to go up, the detector may no longer be in as good a position (relative to the sample) to pick them up. You might simply adjust the insertion of the EDS detector to compensate.
One could use an interpolative process: adjust the tilt a bit, then adjust the detector insertion to maximize counts. Then adjust the tilt a little more, and readjust the detector insertion.
You might get to an optimum geometry faster if you do a rough calculation of the angle of incidence, reflection and best detector insertion to intersect.
Regards, John Best -- ELMDAS Co. http://www.vicon.net/~jbest
} I am looking at a Si wafer with 100nm of SiO2 layer on top. } from a SEM. Electron beam (~8 kev) hit the surface along the } surface normal (0 deg tilt). As I increase the sample tilt } (upto 45 deg or so) EDS quantitative analysis shows a decrease } of O2%. How this is possible? I expected to see O2% goes up } with the tilt as electron will see more of the surface than } bulk. } Dear Fran, What is the geometry (take-off angle, direction of tilt, etc.)? It may be that some O x-rays are taking a longer path and are absorbed, or the Si x-rays can take a shorter path and be absorbed less. Have you used David Joy's Monte Carlo program to calculate the volume in which the x-rays are generated? Yours, Bill Tivol
All else being the same, I (as it sounds you would) would expect an increase in the relative intensity of the oxygen (raw counts). Do you see this on the graphical result? If so, it is most likely that the ZAF/PhiRhoZ/etc. quant.
algorithm is the source of the problem. Woody ______________________________ Reply Separator _________________________________
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Microscopists:
I would like to get an explanation for the following observation of quantitative analysis by EDS.
I am looking at a Si wafer with 100nm of SiO2 layer on top. from a SEM. Electron beam (~8 kev) hit the surface along the surface normal (0 deg tilt). As I increase the sample tilt (upto 45 deg or so) EDS quantitative analysis shows a decrease of O2%. How this is possible? I expected to see O2% goes up with the tilt as electron will see more of the surface than bulk.
Could someone explain why?
Thanks
Fran.
--------------------------------------------------------- Get Your *Web-Based* Free Email at http://www.hotmail.com ---------------------------------------------------------
I teach electron microscopy (both TEM and SEM) as a one semester course. The first couple of times I had 3 to 8 students but last fall I had 14. Students used photographic film and paper out of a common supply. It seemed that every time I turned around, we were out of paper and film or some unidentified person had exposed a portion of our common supply to light. In order to better monitor usage and I am considering giving each student his/her own allotment of paper and film at the beginning of the semester. I require them to turn in 10 TEMs and 10 SEMs at the end of the semester. What do those of you who teach EM (and old fashioned darkroom techniques) require for your students to turn in and how do you budget and/or distribute supplies?
Bob
Robert R. Wise Plant Physiologist and Director, UWO Electron Microscope Facility Department of Biology University of Wisconsin Oshkosh Oshkosh, WI 54901 (414) 424-3404 tel (414) 424-1101 fax wise-at-uwosh.edu
} Dear Microscopists: } } I would like to get an explanation for the following observation } of quantitative analysis by EDS. } } I am looking at a Si wafer with 100nm of SiO2 layer on top. } from a SEM. Electron beam (~8 kev) hit the surface along the } surface normal (0 deg tilt). As I increase the sample tilt } (upto 45 deg or so) EDS quantitative analysis shows a decrease } of O2%. How this is possible? I expected to see O2% goes up } with the tilt as electron will see more of the surface than } bulk. } } Could someone explain why? } } Thanks } } Fran. }
The problem observed here is most likely related to simple rules of geometry. If the EDS detector is not perpendicular to the tilt axis you will find that the path through the sample that the xrays must use to get to the EDS detector becomes longer as the tilt axis increases. (Lower observed take off angle). This results in more absorption. The lower energy peaks will be abosrbed at a greater rate than the higher energy lines. If you are not taking the azimuth angle into your take off angle correction you should.
If you want to limit your xray excitation volume to give you a better idea of the film try also using a lower accelerating voltage. Normally an excitation energy of 2.5 times your xray energy should work.
BTW: I have a DTSA binary -} EMMFF ascii file converter written in Python if anyone needs one. It actually writes a somewhat loose version -- the spec was ambiguous in several places and I loosened some restrictions intentionally for my own purposes. It's written in Python, and it's easily modifiable to your own customizations. ( Specifically, I use longer user defined keyword fields to save the original DTSA header information using the names in the DTSA file interface definition. The DTSA EMMFF plugin seems to read these files back in properly -- the standard tags that is. It ignores the original saved DTSA information. )
I wrote this code before there was a plug-in for DTSA, however I still require it to do batch conversions of folders full of files.
The current version is specifically for the Mac. The initial version was originally run on unix, and most of it should be portable. The main mac specific feature is that it supports drop launching of a whole folder of DTSA files. When I have the final, portable version ( hopefully with a stand-alone Mac version that doesn't require Python installed. ) it will be posted on the Web. If anyone needs it right away, I can email you a copy of the sources.
---| Steven D. Majewski (804-982-0831) {sdm7g-at-Virginia.EDU} |--- ---| Department of Molecular Physiology and Biological Physics |--- ---| University of Virginia Health Sciences Center |--- ---| P.O. Box 10011 Charlottesville, VA 22906-0011 |--- By doing just a little every day, you can gradually let the task completely overwhelm you.
I am a student of EM at Madison Area Tech. We had to buy our own paper. I am now a lot better at making prints !
Leah
---------- } From: wise-at-vaxa.cis.uwosh.edu } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Supplies and teaching } Date: Tuesday, March 04, 1997 9:13 AM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } To all, } } I teach electron microscopy (both TEM and SEM) as a one semester } course. The first couple of times I had 3 to 8 students but last fall I } had 14. Students used photographic film and paper out of a common supply. } It seemed that every time I turned around, we were out of paper and film or } some unidentified person had exposed a portion of our common supply to } light. In order to better monitor usage and I am considering giving each } student his/her own allotment of paper and film at the beginning of the } semester. I require them to turn in 10 TEMs and 10 SEMs at the end of the } semester. } What do those of you who teach EM (and old fashioned darkroom } techniques) require for your students to turn in and how do you budget } and/or distribute supplies? } } Bob } } } Robert R. Wise } Plant Physiologist and Director, UWO Electron Microscope Facility } Department of Biology } University of Wisconsin Oshkosh } Oshkosh, WI 54901 } (414) 424-3404 tel } (414) 424-1101 fax } wise-at-uwosh.edu } }
Greetings, I have a user of our EM Unit who wishes to use Fluoronanogold (Nanoprobes) with Silver enhancement for pre-embedding immuno EM. We will be using frozen 8 micron sections which will then be PLP fixed (parform + Lysine + periodate) prior to labelling. After labelling, we want to post-fix them in OsO4 (as membranes are important), dehydrate and embed in normal epoxy resin. I have heard that the Flouronanogold is not stable at 60oC, the normal curing temperature of the resin.
Has anybody had experience with pre-embedding with Fluoronanogold? Will Fluoronanogold survive the epoxy resin curing process at 60oC? If it wont survive, which resin and curing procedure would be the choice to use in this case? (Quetol 651, Lowicryl?)
I am interested to replace our SEM by ESEM or low vacuum SEM, but I would like to have more information about EDX microanalysis with these SEM (what are the quantitative and qualitative differences when I change the pressure? what's happen with cryo system .....?). Thanks in advance for your help (comments, references ...)
Best regards to all Didier
-------------------------------------------- Dr. Didier Le Thiec I.N.R.A. Centre de Recherches Forestieres Unite d'Ecophysiologie Forestiere Laboratoire de Pollution Atmospherique 54280 Champenoux - France
First, my apologies to the Microscopy listserver subscribers if this posting isn't appropriate. We need a first rate science teacher with experience, and my colleagues here at the microscopy listserver sprung to mind immediately.
Secondly, If your not interested in accepting a position as a high school science teacher, and know of a listserver that may produce some interested candidates, please forward this request. Thank You.
And finally: The Juniata Valley School District is seeking candidates for the position of Science Teacher beginning in the fall of 1997. Responsibilities are: fundamentals of science and physical sciences for students in grades 7-12.
We're looking for candidates who bring a number of years of experience from industry or a university research environment to the classroom. Pennsylvania teaching certification is required, or must be readily obtainable. The ability to motivate and educate "typical" high school students is a must.
With a BS the starting salarary is $28,041, an MS starts at $28,712. Starting salaries are dependant on teaching experience within the Pennsylvania public school system.
The Juniata Valley School District has approximately 500 students in grades 7-12 and is situated in a beautiful rural area approximately 25 miles southwest of the main campus of The Pennsylvania State University. JVSD is an equal opportunity employer.
Please forward your resume to our superintendant, Dr. David Leckvarcik at The Juniata Valley School District, P.O. Box 318, Alexandria, PA 16611. If you would like me to follow up on your resume, please forward it to me at: jbest-at-vicon.net.
I am wondering what solutions people have found to the stage temperature/gas/humidity control problem for tissue culture microscopy of live cells. I have tried an open chamber approach (20/20 Technologies) and evaporation onto the condenser or lid was a major problem as was even heating. Are there any commercial stage temperature controllers that rely on hooding the microscope or stage that work well? Has anyone constructed something simple that can be reproduced easily? What are the down sides of this approach? Has anyone used the new Bioptechs system and does it actually do the job and solve any or all of these problems? Thanks- Dave
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut U-125 Storrs, CT 06269 Knecht-at-uconnvm.uconn.edu
I am looking for a PC based software (preferably on Windows) that can perform profilometry type measurements from an SEM pair of images (grabbed into 'TIF' files). Measurements should include: - Line profiles - Height measurements - Contour maps.
Any help is appreciated.
Regards,
Dr. Rafik Allem Pulp and Paper Research Institute of Canada. 570, St. John's Boulevard, Pointe-Claire, Qc, H9R 3J9, Canada. tel: (514)630-4101 ext. 2661, fax:(514)630-4134 e-mail: allem-at-paprican.ca
Department of Mining and Metallurgical Engineering Professional Development Seminars
"MATERIALS CHARACTERIZATION FOR THE METAL, MINERAL AND MICROELECTRONICS INDUSTRY" May 12-15, 1997
Seminar Leader: Neil Rowlands
This course is intended to benefit users who are concerned with the analysis of fine structures, fine particles and may also have the need to image surfaces down to the atomic level. These techniques will be of special interest to those involved in the microelectronics, polymer science, and ceramics industries, as well as those involved in metallurgical analysis.
The course will be of a practical nature and will be of interest to operators and managers of analytical laboratories R&D engineers and applied scientists. Examples of the applications of each technique will be presented and on the last day of the course there will be an optional visit to the Materials Technology Laboratory, CANMET in Ottawa to see many of these techniques in operation. Additional facilities (e.g. TEM, AFM) are available on the McGill campus.
I already replied privately before I got down our in-box to the MSA posting. For anyone interested in this, here are our ruminations...
Generally, we recommend low-temperature polymerization resins such as Lowicryl, LR White or any similar medium for embedding Nanogold=81 or =46luoroNanogold-labeled specimens. This is because some early experiments with Nanogold=81-labeled specimens suggested that the higher-temperature embedding (60=B0C) might cause the gold particles to be displaced from their binding sites. However, subsequent experiments with Nanogold=81 in solution showed that it could be heated even up to 100=B0C for 1 h with minimal decrease in optical density; generally, avoiding low pH values (below 7) or high ionic strengths (0.2 M NaCl or higher) helps ensure its stability. Because it is smaller than most colloidal gold and has no tendency to stick electrostatically to proteins or cell components, it may be more free to move: fixing with glutaraldehyde helps counteract this, and with Nanogold=81 we also recommend silver enhancement before embedding if it is practical.
There is a section on this in Hainfeld and Furuya's chapter on the silver enhancement of Nanogold=81 and undecagold in M. A. Hayat's recent book, "Immunogold-Silver Staining: Principles, Methods and Applications" (CRC Press, Boca Raton, 1995; pp. 71-96. Check pages 92-92 for the effects of heating Nanogold=81. From this section, heating at 60=B0C for 250 minutes resulted in a reduction of the optical density to 80% of its initial value - suggesting that Nanogold=81 can survive most 60=B0C embedding procedures.
The gold particle in FluoroNanogold is Nanogold, and exactly the same applies to this probe. However, silver enhancement quickly removes the fluorescence, so only do the pre-embedding silver enhancement if you have completed the fluorescence microscopy.
We keep a list of answers and suggestions to frequently-asked questions in the Technical Help section of our web site:
http://www.nanoprobes.com/Tech.html
All the suggestions made there about Nanogold=81 use and stability also appl= y to Fluoronanogold, since these probes use the same gold particle.
Hope this is helpful,
Rick Powell
****************************************************************** * NANOPROBES, Incorporated | Tel: (516) 444-8815 * * 25 East Loop Road, Suite 124 | Fax: (516) 444-8816 * * Stony Brook, NY 11790-3350, USA | nano-at-mail.lihti.org * * * * NOW EASY TO FIND ON THE WEB: http://www.nanoprobes.com * ******************************************************************
In response to Allan Mitchell's inquiry. If you are speaking of whether the Fluorescent signal of Fluoro-Nanogoold survives the answer is no, but not because of the 60 degree heat. The fluorescence is quenched or covered up by the silver enhancement process. If you want to see the label prior to silver enhancement that is no problem. I often do this prior to running the silver enhancement to see that the reagent has gotten in to the sample and gives the expected signal. Your other choice of resins are lowicryl which you can polymerize at -35 degrees under UV. The FLUOR tag survives this just fine. In regard to osmication, this has to be done with 0.1% osmium at 4 degrees or on ice for about 30 - 45 minutes. Standard osmication procedures will reduce the silver shell that forms around the nanogold particle. This appears as a much less electron dense cloud or ghost. I have used these reagents with spurr, epox 812, lowicryl and LR White without any problem.
Joe Goodhouse Confocal / E.M. Lab Molecular Biology Princeton University
} From: wise-at-vaxa.cis.uwosh.edu } Date: Tue, 04 Mar 1997 09:13:27 +0000 } Subject: Supplies and teaching } 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 } -----------------------------------------------------------------------. } } To all, } } I teach electron microscopy (both TEM and SEM) as a one semester } course. The first couple of times I had 3 to 8 students but last fall I } had 14. Students used photographic film and paper out of a common supply. } It seemed that every time I turned around, we were out of paper and film or } some unidentified person had exposed a portion of our common supply to } light. In order to better monitor usage and I am considering giving each } student his/her own allotment of paper and film at the beginning of the } semester. I require them to turn in 10 TEMs and 10 SEMs at the end of the } semester. } What do those of you who teach EM (and old fashioned darkroom } techniques) require for your students to turn in and how do you budget } and/or distribute supplies? } } Bob } } } Robert R. Wise } Plant Physiologist and Director, UWO Electron Microscope Facility } Department of Biology } University of Wisconsin Oshkosh } Oshkosh, WI 54901 } (414) 424-3404 tel } (414) 424-1101 fax } wise-at-uwosh.edu } } } Bob,
We here at BYU teach a course in SEM and in TEM. We require our students to submit two types of assignments to complete the course. First is a portfolio which may or may not use traditional darkroom techniques (digitized, laser printed images are an option from our SEMs). We also have the students turn in a poster representing a project they have worked on all semester. The poster must have at least 5 high quality photographs which the student has produced, as well as any text and graphics the student wants to display. The student purchases all of the photographic materials they use. We do keep a stock supply in the lab for the students to purchase from. Students are responsible for all of their own photographic supplies so if thier paper becomes light struck, that is their problem. This has worked quite successfully for the past several years.
I hope this helps
Michael D. Standing e-mail: MDStandi-at-bioag.byu.edu Phone: (801)378-4011
I have noted several users of this listserver refer to the Ilford RC2150 print processor. We have used one for several years without trouble but Ilford has apparently switched to a new "environment friendly" cleaning solution that is unable to remove the deposits from the rollers. All of our prints have terrible streaks of silver residue. Has anybody else had this problem - more importantly, has any been else solved it? TIA
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
Way back when, the TEM class I took, it was required for me to purschase my own film, paper, tweezers, embedding mold and a text book. All this was setup by the proffessor and sold thru the bookstore on campus. Yes it was/is expensive but it taught me to be conservative with these items. Most of the prints I made for the class were 4x5. The class was ten weeks long and I did not run out of supplies.
Hopes this helps,
Ed Calomeni Dept. Pathology Medical College of Ohio Toledo, Ohio emlab-at-opus.mco.edu
I am trying to use stains discribed in Histochemistry: Theoretical and Applied Volume 2 by AGE Pearse to locate metallic ions with in tissue. Any suggestions for other methods will be appreciated. The following are the stains, the metal I'm trying to detect, and the problem I am having. I trying to stain cells not sections.
Napthochrome Green B Beryllium (1)unable to find acridine red Aldridge says is was discontinued 12 years ago
Rubeanic Acid Nickel (1) the stain is very light and is not staining cells
Chrome Azurol S Chromium (1)library is unable to get referenced paper and therefore I am unable to find protocol
If anyone has any information that might help please responed.
Regina Messer Graduate Student Biomedical Engineering UAB
For staining metals look in "Histopathologic Technique and Practical Histochemistry" by Ralph Lillie and Harold Fullmer; McGraw Hill, 1976. Out of print for years but probable available in your library or via interlibrary loan. Also "Selected Histopathological Methods" or something like that by Thompson is a massive tome with just about everything in it. Call or e-mail if necessary.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (908)-235-4583; fax -4029 e-mail: mcauliff-at-.umdnj.edu ***************************************************************
We have observed that spatial x-ray resolution suffers, even at 40 Pa with a 25 mm working distance. We can get decent x-ray maps, but quantitative analyses can easily pick up tenths of a percent of an element from a nearby phase. We have yet to quantify the effects for ourselves. I recall there were a number of papers at the Minneapolis meeting on this effect. One has to be careful.
At 10:30 AM 3/5/97 -0500, you wrote:
} Dear all, } } I am interested to replace our SEM by ESEM or low vacuum SEM, but I } would like to have more information about EDX microanalysis with these SEM } (what are the quantitative and qualitative differences when I change the } pressure? what's happen with cryo system .....?). } Thanks in advance for your help (comments, references ...) } } Best regards to all } Didier } } -------------------------------------------- } Dr. Didier Le Thiec ---------------------------------------------------- Warren E. Straszheim 270 Metals Development, Ames Lab/ISU, Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-3091
Hello: I am looking for a used SEM with low Voltage capability, about 1 KV. Let me know if you know of anybody who would like to sell. Please E-mail or call me at 909 694-1839. Thanx, Peter Jordan
Didier Le Thiec wrote: } I am interested to replace our SEM by ESEM or low vacuum SEM, but I } would like to have more information about EDX microanalysis with } these SEM (what are the quantitative and qualitative differences when } I change the pressure? what's happen with cryo system.....?)
Dear Didier,
The presence of gas in the specimen chamber influences the spatial resolution of EDX microanalysis in ESEM and LVSEM. The primary electrons will scatter on the gas and give rise to a skirt of scattered electrons that will excite X-rays at places pretty far away from the point you want to analyze. The scattered intensity is approximately given by Is/Io = 1 - exp(-psL/kT) where p is the pressure, s the total scattering cross section for electron scattering on the gas used, L the distance between the last pressure limiting aperture and the sample, k the Boltzmann constant and T the absolute temperature. Examples of skirt shapes are given in:
D. A. Moncrieff et al., J. Phys. D: Appl. Phys. vol. 12 (1979) 481-88. D. C. Joy, Microscopy and Microanalysis ' 96, Proc. Annual Meeting MSA, Minneapolis, Minnesota, 11 - 15 August 1996
It is to a large degree possible to correct for the beam skirt effects:
1) You can extrapolate from spectral measurements made at several different pressures to the result that would have been found without scattering provided that the measurements are made in the single scattering regime (e.g. pL { approx. 1.6 Pa.m for measurements in water vapour). 2) If there is plural scattering, you can take two spectra, one with a fine needle (of the kind used for field ion microscopy or scanning tunneling microscopy) inserted over the point of interest, and the other with the needle slightly retracted. Subtraction of the first from the second spectrum will approximately give the spectrum from the point of interest.
Neither method will give you as exact an analysis as you will get under high vacuum, but you can get rid of most of the skirt effects. The pressure variation method in particular yields pretty good results if carefully performed. The methods are described in:
J. B. Bilde-Soerensen and C. C. Appel, Proc. 48th Annual Meeting of the Scandinavian Society for Electron Microscopy, Aarhus, 2 - 5 June 1996, pp. 4 - 5. J. B. Bilde-Soerensen and C. C. Appel, Proc. 11th European Congress on Microscopy EUREM' 96, Dublin, 26-30 August 1996. Session T6.
ESEMs and LVSEMs can also be operated as high vacuum SEMs - so you still have the possibility of going to high vacuum for microanalysis.
Best wishes, Jorgen. -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- J. B. Bilde-Soerensen Materials Research Department Risoe National Laboratory DK-4000 Roskilde Denmark
We have published a couple of papers on ESEM EDX. Might be worth a look. :) I would like to add that if you wish to look at hydrated samples then the chamber pressures required are such that scattering of the primary beam results in a probe at the sample of at least 1mm diameter!! Also correction methods based on taking spectra at different pressures are fraught with difficulty if you need to avoid drying out your sample. Also be aware that with low Z detectors a contribution from the chamber gas will be encountered.
Chris
} I am interested to replace our SEM by ESEM or low vacuum SEM, but I } would like to have more information about EDX microanalysis with these SEM } (what are the quantitative and qualitative differences when I change the } pressure? what's happen with cryo system .....?). } Thanks in advance for your help (comments, references ...) } } Chris Gilpin Biological Sciences Electron Microscope Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 161 275 5170 fax +44 161 275 5171
At 02:43 PM 3/5/97, Hildagonda van der Merwe wrote: } Can anyone please tell me all the uses of the Immuno-Bed Kit?
Immunobed is an embedding medium for immunohistochemistry which allows the rapid penetration of large immunoglobulins through the section for demonstration of antigenic sites.
For more detailed information, please request our A2050 data sheet.
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
I wish to thank those people (Scott Whittaker, Gary Chinga, Christian Mellen, Melvyn Dickson, John Mardinly, Hans Tietz) who replied me, through there were a few. (Sprinkling water seems to be much more interesting to talk about.:) Short summary is: The main software for image analysis is NIH (FTP from zippy.nimh.nih.gov), through analySIS also exists and Photoshop, Paintshop, Coreldraw and Wincatpro can be used to manipulate images.
CD-Rs mainly used to store, transport and distribute image files.
1200dpi LP (mainly Lexmark) used for draft and dye-sub printers for quality prints. So one will need both...
Useful references obtained: http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks " site supported by Scott Whittaker - many discussions from this Listserver and Confocal Listserver archived there. Extremely useful!
http://www.soft-imaging-web.de home of Soft-Imaging Software information about analiSIS software package, etc.
Stewart, M.G. and Davies, H.A. (1995) Digital image processing (DIP) in the TEM: is it viable in biological morphometry?. Eur Microsc Anal (1995) 35, pp 21-23.
Special opinion: "Andrej! Do not change!..."
Full text of replies can be obtained on request. A. Chuvilin
I saw your concern about the EDX at higher pressures. As is suggested by others there are inaccuracies in the quantitative estimation. This is because the incident beam coming through the gaseous region spreads before it hits the specimen surface. The electrons outside the original gaussian profile of the beam also create x-rays. You are probably aware of David Joy's work which was presented at MSA meeting in the US. We are enormously interested in this problem and one of my students is doing Monte carlo calculations on this very problem. You also inquired about cryoSEM. Normally, you should not have any problem about the beam scattering as the equilibrium vapour pressure is below the chamber pressure. So you can assume that all of your x-rays are coming from the original focussed beam spot. This of course assumes that you are not using a very intense beam which evaporates water or other constituents in your specimen and upsetting the inherent composition of the specimen. Incidentally,at Bristol we convert conventional SEMs in to High pressure SEMs exceeding the performance of the commercially avilable instruments. If you need any help on this please do not hesitate to get in touch with me
Yours sincerely
Jitu Shah
Dr.Jitu Shah H.H. Wills Physics Laboratory, University of Bristol, Royal Fort, Tyndall Avenue, Bristol BS8 1TL. UK email: jss-at-siva.bristol.ac.uk Tel: 44 117 9288719 Fax: 44 117 9255624
When I perform an EDX analysis on uncoated fused silica, 10 20 or 30 KeV, and the sample begins to charge, often I get a small aluminum peak, and it doesn't go away as I move around the sample, it comes back. The amount of aluminum in the quartz is typically less than 20 ppm, so I don't think that the Al peak is coming from my sample. Could it be that the Al peak is coming from my sample stage or walls of the sample chamber, due to any effect caused by the charge that has developed on the uncoated quartz? Also, for EDX work, what are the negative effects that sample charging has on an analysis? My work is done on an Amray 1600 with a Noran detector, one that has two windows, Be and the ultra thin window.
Recently I got a problem in LVSEM-EDS system. The SEM is JSM-5400LV with Kevex EDS. We use a thin B-window with expect to determine elements down to Z=5. The SEM does not have pre-vacuum chamber which means whenever we change sample, the chamber and whole column are exposured to atmosphere. The window has been used for 10 months with good performance. Therefore, recently once we change sample as usual, the window got brocken. We have no idea what is the reason.
My question is: How thick window should I use to prevent the window from brocken, and what is the common technical hints to prevent the window from brocken? Does the frequently presure change on window surface (when change sample) affect the life time of window?
Thank you very much
Zhiyu Wang Electron Microscope Facility Western Kentucky University Bowling Green KY 42103 USA wangz-at-pulsar.cs.wku.edu
I am trying to do trace Carbon analysis in Nb material. The Be window of my EDX can be opened in order to detect C, N, O, but the quantification seems impossible. Anybody has similar experience?
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello Everyone,
I have been tasked with bringing another technician into our SEM lab and would like to get info on the courses which are available for a person who is familiar with light microscopy but knows almost nothing about a SEM.
The desired course would be 5 days or less and be applicable to the level of knowledge described above. We deal with strictly materials analysis, so biological based courses are not applicable. The desired outcome would be someone who has a rudimentary knowledge of SEM theory and capable of taking basic SEM pictures and acquiring EDS spectra on non-challenging samples. We will be doing some training of the new tech, but due to heavy workloads, we can't devote the time to the training that we would normally do.
Geographically, I would like courses in the Southeastern USA, but will take info on courses anywhere in the US.
John Giles Senior Materials Engineer Honeywell Space Systems Clearwater, FL
The EM facility at EPA-Cincinnati is hopefully going to replace a 20 year old critical point dryer in the near future. If anyone has any recommendations, or horror stories, let us know before we spend your tax dollars. You can reply to us directly at clark.patrick-at-epamail.epa.gov Thanks
This happened to me once, and it turned out to be stray x-rays from the EDX detector collimator sleeve. If your detector has one of these, slide it off and paint the inside surfaces with carbon dag and try it again; it may solve your problem.
Regards,
-Bob ********************************** Bob Citron Chiron Vision 555 W. Arrow Hwy Claremont, CA 91711 ph: (909)399-1311 email: Bob_Citron-at-cc.chiron.com **********************************
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
When I perform an EDX analysis on uncoated fused silica, 10 20 or 30 KeV, and the sample begins to charge, often I get a small aluminum peak, and it doesn't go away as I move around the sample, it comes back. The amount of aluminum in the quartz is typically less than 20 ppm, so I don't think that the Al peak is coming from my sample. Could it be that the Al peak is coming from my sample stage or walls of the sample chamber, due to any effect caused by the charge that has developed on the uncoated quartz? Also, for EDX work, what are the negative effects that sample charging has on an analysis? My work is done on an Amray 1600 with a Noran detector, one that has two windows, Be and the ultra thin window.
Would you be so kind as to cite your literature references, so we can read them. They sound useful for a number of us doing EDX in a variable pressure SEM. Thanks.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 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 ####################################################################
Looking for image analysis software/solutions for the following:
1} sperm analysis,cell motility etc. 2} textile analysis,rayon fibre analysis,broken fibre etc.
Could anyone point out some good manufacturers/sources with address/email/fax wwith name of source.?////// I would be grateful if someone could help me in this context.
Thanks,
Best regards,
Anish
************************************************************************* For further details please contact: Soneja A.K. Director METZER BIOMEDICAL & ELECTRONICS PVT.LTD. 327 Wadala Udyog Bhavan,Wadala,MUMBAI(BOMBAY )400 031.INDIA Tel:91 22 4145057/4165650 Fax 91 22 4168757
There are a couple of reasons why windows can have a limited lifetime when used in LV-SEM & ESEM.
Firstly, when the chamber is pumped to and from atmosphere any loose particles in the chamber are disturbed, and can damage the window by hitting it. This can be reduced by fitting some kind of loose cover (we use a plastic bag!) over the x-ray probe whenever using the microscope for non x-ray work.
Secondly, certain windows when used in lo-vac are susceptible to ice formation on the surface and actually in the window.
Regarding probe-beam spreading, we, like Dr. Shah, are carrying out monte-carlo simulations, in conjunction with x-ray measurements and will hopefully be presenting some results at this years MSA meeting
Ian Bache Cavendish Laboratory Cambridge University Madingley Road Cambridge ENGLAND (+44)-1223-337229
} Would you be so kind as to cite your literature references, so we can read } them. } They sound useful for a number of us doing EDX in a variable pressure SEM. } Thanks. } Sorry I should have included them on the original post!
Sigee, D.C. and Gilpin, C.J. X-ray microanalysis with the environmental scanning electron micrasocpe: Interpretation of data obtained under different atmospheric conditions. Scanning microscopy supplemnet 8 219-229 1994
X-ray microanalysis of wet biological specimens in the environmental scanning electron microscope: 1 Reduction of specimen distance under different atmospheric conditions. Journal of Microscopy 179 (1) 22-28 1995
More work in progress as they say!
Chris
Chris Gilpin Biological Sciences Electron Microscope Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 161 275 5170 fax +44 161 275 5171
} Would you be so kind as to cite your literature references, so we can read } them. } They sound useful for a number of us doing EDX in a variable pressure SEM. } Thanks. } Sorry I should have included them on the original post!
Sigee, D.C. and Gilpin, C.J. X-ray microanalysis with the environmental scanning electron micrasocpe: Interpretation of data obtained under different atmospheric conditions. Scanning microscopy supplemnet 8 219-229 1994
X-ray microanalysis of wet biological specimens in the environmental scanning electron microscope: 1 Reduction of specimen distance under different atmospheric conditions. Journal of Microscopy 179 (1) 22-28 1995
More work in progress as they say!
Chris
Chris Gilpin Biological Sciences Electron Microscope Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 161 275 5170 fax +44 161 275 5171
John Giles wrote: ===============================================
I have been tasked with bringing another technician into our SEM lab and would like to get info on the courses which are available for a person who is familiar with light microscopy but knows almost nothing about a SEM.
The desired course would be 5 days or less and be applicable to the level of knowledge described above. We deal with strictly materials analysis, so biological based courses are not applicable. The desired outcome would be someone who has a rudimentary knowledge of SEM theory and capable of taking basic SEM pictures and acquiring EDS spectra on non-challenging samples. We will be doing some training of the new tech, but due to heavy workloads, we can't devote the time to the training that we would normally do. ================================================= We would give extremely high marks to the "Lehigh" courses which are given at Lehigh University, Bethlehem, PA every year. You can get information about their program from their website at the following:
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
We anticipated window problems from a detector left in the wet environment 24/7 and cycled for every sample change. We initially had a hard time finding an EDS vendor to build us a detector that was fully retractable behind a gate valve outside the 2020 chamber so that the detector was protected not only from projectiles and cycles but from harsh environments sometimes used in environmental scopes. We did eventually get such a detector and had window problems initially but that seems to have been solved with a slightly thicker window. My advice is that everyone should let their EDS reps know that this type of detector is desirable and maybe one day they will be widely available.
I also wonder if the commercially available window materials are water tight. As we all know one of the EDS vendors uses a heater to remove ice from its crystal. Helium leak tests are often quoted when asked about water permeability but I would like to see water leak test data. If water is geting into and through windows it could be responsible for lost windows in wet systems.
I am looking forward to these monte carlo papers and lively discussions at the M&M meeting in Clevland.
Scott Wight
} There are a couple of reasons why windows can have a limited lifetime when } used in LV-SEM & ESEM. } } Firstly, when the chamber is pumped to and from atmosphere any loose } particles in the chamber are disturbed, and can damage the window by } hitting it. This can be reduced by fitting some kind of loose cover (we } use a plastic bag!) over the x-ray probe whenever using the microscope for } non x-ray work. } } Secondly, certain windows when used in lo-vac are susceptible to ice } formation on the surface and actually in the window. } } Regarding probe-beam spreading, we, like Dr. Shah, are carrying out } monte-carlo simulations, in conjunction with x-ray measurements and will } hopefully be presenting some results at this years MSA meeting } } } } Ian Bache } Cavendish Laboratory } Cambridge University } Madingley Road } Cambridge } ENGLAND } (+44)-1223-337229
------------------------------------------------------------------ Scott Wight e-mail: SCOTT.WIGHT-at-NIST.GOV NIST - Microanalysis Group W voice: 301-975-3949 Bld 222, Rm A113 | fax:301-216-1134/301-417-1321 Gaithersburg, MD 20899 \|/ disclaimer: Any opinion expressed is my own and does not represent those of my employer.
Try Visilog from Noesis Vision Inc, we have a turn key system available for sperm analysis and or fibre analysis. You can reach us at http://www.noesisvision.com
At 12:51 PM 3/7/97 +0530, SONEJA A K wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
---------------------------------------------------------------------------- --------------------- Luc Nocente Tel: 514 345 1400 Noesis Vision Inc. Fax: 514 345 1575 e-mail: ln-at-noesisvision.com 6800 Cote de Liesse, Suite 200 St-Laurent, PQ H4T 2A7,Canada
Visit our new web site at http://www.noesisvision.com ---------------------------------------------------------------------------- ---------------------
The Royal Microscopical Society Computers in Microscopy ,September 1997, Cambridge CALL FOR PAPERS
A Colloquium on various aspects of the application of computers to microscopes and microscopical techniques is being planned by the Royal Microscopical Society, to be held in the University Engineering Department, Trumpington Street, Cambridge, on Thursday 25th September 1997.
The colloquium is intended to cover recent progress made in the use of computers for the assessment, interpretation, restoration and enhancement of microscopical images, including, but not restricted to applications in optical and all kinds of electron microscopy. Of particular interest will be contributions concerning new approaches and techniques including 3D measurements and profiling, wavelets and fractals. It is hoped that papers will cover some of the comparatively new fields of application, including multi-channel imaging, instrumental control and remote microscopy. It is proposed also to organise a small exhibition of products and materials by local organisations involved in this area, as well as a visit to laboratories in the University engaged in this kind of work.
The colloquium takes place immediately after a three-day course, also entitled 'Computers in Microscopy', and it is intended that the two events be complementary. ( Please note that it is not necessary to attend the course in order to register for the colloquium.)
If you would like further information about the Colloquium and/or the Course please contact Rebecca Morden either by email: info-at-rms.org.uk, by telephone: +44 (0)1865 248768 or by fax: +44 (0)1865 791237.
Prospective contributors are invited to submit a synopsis of approximately 150-200 words, before 30 April 1997, to Dr D M Holburn, University Engineering Department, Trumpington Street, Cambridge CB2 1PZ, U.K. Enquiries may be made by electronic mail, to dmh-at-eng.cam.ac.uk, by facsimile to +44 1223 332662, or by telephone to +44 1223 332775.
******************************************************************************* Rebecca Morden Course Organiser Royal Microscopical Society, 37/38 St Clements, Oxford, OX4 1AJ, UK. Tel (0)1865 248768, Fax (0)1865 791237, email info-at-rms.org.uk *******************************************************************************
Thank you very much for providing information and experience in window breackage problem. I got windows replace service with the price of $6500 (does not including freight charges). We do not know how to handle this problem if the window does not stand in a reasonable lifetime or unlucky, got several window breackages. It must be a big meney gone! Is it possible to find a cheaper service? Any suggestion?
I have been tasked with bringing another technician into our SEM lab and would like to get info on the courses which are available for a person who is familiar with light microscopy but knows almost nothing about a SEM.
The desired course would be 5 days or less and be applicable to the level of knowledge described above. We deal with strictly materials analysis, so biological based courses are not applicable. The desired outcome would be someone who has a rudimentary knowledge of SEM theory and capable of taking basic SEM pictures and acquiring EDS spectra on non-challenging samples. We will be doing some training of the new tech, but due to heavy workloads, we can't devote the time to the training that we would normally do.
Geographically, I would like courses in the Southeastern USA, but will take info on courses anywhere in the US.
John Giles Senior Materials Engineer Honeywell Space Systems Clearwater, FL
} John Giles wrote:-----------------------------------------------. } } Hello Everyone, } } I have been tasked with bringing another technician into our SEM lab and would } like to get info on the courses which are available for a person who is } familiar with light microscopy but knows almost nothing about a SEM. } } The desired course would be 5 days or less and be applicable to the level of } knowledge described above. We deal with strictly materials analysis, so } biological based courses are not applicable. The desired outcome would be } someone who has a rudimentary knowledge of SEM theory and capable of taking } basic SEM pictures and acquiring EDS spectra on non-challenging samples. We } will be doing some training of the new tech, but due to heavy workloads, we } can't devote the time to the training that we would normally do. } } Geographically, I would like courses in the Southeastern USA, but will take } info on courses anywhere in the US. } } John Giles } Senior Materials Engineer } Honeywell Space Systems } Clearwater, FL
John:
A number of us run an SEM course at the University of Maryland called 'Practicle Aspects of SEM. This course is a 4 1/2 day course covering the basic theory and practice of SEM (including basic x-ray microanalysis) with direct hands-on experience for all students. In addition, students are encouraged to bring samples of their own. This year the courses are set for May 19-23 and May 26-30. If you would like more information please feel free to contact either me ( 315-859-4715) or Tim Maugel (301-405-6898)
Ken Bart
Kenneth M. Bart Director, Electron Microscopy Facility Hamilton College Clinton, New York 13323 USA kbart-at-hamilton.edu (315) 859-4715
We are in need of a vibration isolation pad for an Auger spectrometer, which has the dimensions of a standard SEM. Can anybody recommend a manufacturer or can recommend homemade solutions?
We have a WDS system for sale. The system consists of the computer, software, spectrometer controls and stage controls, NO DETECTORS or SPECTROMETERS are included.
It is a Kevex Delta 4 / Sesame system (type 8006) with a 5724-004 console, containing 4 spectrometers controls, 3 stage motors, and 4 spectrometer pre-amps and ratemeters, beam meter. A Tectronix plotter and a matrix printer a part of the package.
The system was configured to run on a JEOL JXA 8600 superprobe.
Price: $ 5000, you pay freight.
Hasso Weiland Alcoa Technical Center Alcoa Center, PA 15069
I hope this doesn't seem out of palce on this list, but it is equipment I need to print spectra and images from ny TEM work. Has anyone had to replace the printhead on an Apple Color StyleWriter Pro and gotten a better price than the going Apple rate of $250! (new printer time?) Alternately, has anyone got a unit that was junked for other reasons that might have a usable printhead that they would be willing to sell/donate? The one that I use in my electron microscope lab had a malfunction which forced ink on top of the printhead and corroded out the contacts. Thanks in advance for any help/advice you can offer.
Sincerely, Andy Andy Buechele The Catholic University of America 409 Hannan Hall Washington, D.C. 20064 (202) 319-4995 FAX: (202) 319-4469
Y'all are an encyclopedia of knowledge, and I want to check out a volume... I have just recently been learning SEM and all the associated procedures that precede going on the scope. I have had to learn this all on my own, which is why I'm having this problem. When I do a cpd on a sample they always come out looking wrinkled. Am I going wrong in the cpd or is it some other step. I've heard that some people do a few purges of the system and some do a lot. I've just purged the ETOH until I see CO2 snowballs & then let it sit for 15 min. & then repeat the cycle 2 more times. Is this wrong? Can someone out there give me some pointers? I'm getting pretty tired of my stuff looking like raisins all the time. = (
Thanks in advance!
Paula = )
Paula Ssicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
This seems to be a question of general interest. As the manufacturer of the majority of ultrathin windows, MOXTEK can offer the following suggestions to improve window lifetimes.
1) Slow down the chamber venting. The most common field failure is what we call "bullet holes." These occur when particulate in the chamber is "swirled up" during venting and impacts the surface of the window.
2) Retract the detector before you vent, if you have this capability. (See #1 for the reason.) I saw an earlier suggestion to cover the detector with a plastic bag when it is not in use to minimize the exposure to this flying particulate. Good suggestion!
3) Never pump down your specimen chamber with a warm x-ray detector attached. Make sure you cool down the detector before pumpdown. Most of the detector systems have a molecular sieve or other getter material in the dewar vacuum to trap any residual gases. When you warm up the detector this material can outgas the trapped gases. If you then pump down the specimen chamber you reverse pressure the window. These windows are not designed to tolerate any back pressure.
4) Never overheat the window by exposure to a hot stage. Your instrument manufacturer should be able to recommend how to operate the system to minimize heat transfer to the detector window.
5) Consult with your EDX manufacturer for specific instructions for your system. Every detector/microscope combination is unique, and they should have some experience that will help you.
There are various thicknesses of windows available, but the above suggestions apply regardless of the thickness. If you continue to have failures with the thinnest windows, a thicker option is probably available through your EDX manufacturer.
(Please note that the Kevex B windows are unique to Kevex, so you should probably talk to them about the problem you had. With a silicon support grid like the MOXTEK ultrathin windows there is no problem in frequent pressure changes breaking the window.)
Hope this helps.
D. Clark Turner Director MOXTEK, Inc. 452 West 1260 North Orem, Utah 84057 phone (801) 225-0930 fax (801) 221-1121 email moxtek-at-moxtek.win.net
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
REGARDING Job Posting
POSITION OPEN: Postdoctoral Scholar or Research Associate
Area: Materials Science/Analytical Electron Microscopy/Lorentz Imaging
Qualification: A PhD in Materials Science/Physics or related area. Very strong hands-on experience in various TEM techniques and their application to microstructural characterization at high resolution is required. Experience in Lorentz Imaging and TEM specimen preparation is desirable. The suitable candidate will be comfortable using the wide range of TEM instrumentation available at the NCEM. This post-doctoral fellow will spend a major fraction of the time working on the microstructural characterization of plastically deformed samples. The goal is to explore the correlation between the local physical/chemical microstructure in plastically deformed regions with local changes in magnetic structure(measured independently by SQUID imaging techniques). The remaining time will be devoted to the development of Lorentz Imaging techniques on our new Philips 200KV/FEG instrument using a variety of thin film samples.
The position is open immediately for at least one year, renewable upon mutual agreement for longer period, provided funds are available. Salary and benefits will commensurate with experience and skills.
For more information on the scientific/technical aspects of the position please contact :
Kannan Krishnan 72-209, NCEM Lawrence Berkeley National Laboratory Berkeley, CA 94720
Lehigh University, Bethlehem, PA has beginner as well as advanced SEM/EDS (+other) courses. These run for a week, and are well done. Sorry, I tossed my mailing about the course so I am missing the details. Believe it is scheduled for June. I am sure someone else will jump in with the details.
There are other (similar) courses available, but I have not experienced those....
Quantitative (even standardized) EDS analysis for carbon in Nb is difficult to impossible, no matter what the EDS manufacurers may imply. The same applies for carbon in Zr. Several things..... 1) Very light element (soft x-rays) in a high Z matrix. 2) Nb is a strong absorber of carbon x-rays. 3) Since the matrix carbon signal is weak, any surface carbon (contamination) can be a major contributor to the carbon peak. Together, these make for very large absorption correction coefficients and magnify surface contamination contributions. Translate that: error.
To maximize your chances....
Don't rely on standardless (semi) quant.
A clean, high vacuum is required to minimize surface (carbon) during analysis.
Specimen surface preparation is crucial. A flat, well polished, and very clean specimen surface must be achieved.
Do not coat the specimen (NbC is sufficiently conductive anyway).
Minimize analysis volume and depth. - (A) Use as low a beam voltage as practical. (B) High tilt angles will skew the volume to near surface. Caveats: (A) Nb, L-line (family) response less predictable than Ka line. I have run across some quant software which won't use L-lines- at least for stdless quant. (B) Less than perfect quant compensation for high tilt may add some error...(especially for stdless).
For stdless quant, I have seen demos produce errors } 300 percent.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am trying to do trace Carbon analysis in Nb material. The Be window of my EDX can be opened in order to detect C, N, O, but the quantification seems impossible. Anybody has similar experience?
Message-Id: {2.2.32.19970308041320.006ac864-at-pop.unixg.ubc.ca} X-Sender: mager-at-pop.unixg.ubc.ca X-Mailer: Windows Eudora Pro Version 2.2 (32) Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Dear Hasso, I have had some success reducing the vibration evident on my SEM by making four pads from Sorbathane. I used a fairly stiff number of Sorbathane about 3/8 inch thick and cut four pads about four inches square. The shop made me eight steel plates to go on either side of the Sorbathane and I put these under the corners of the column section. The vibration on my SEM (from being on the fourth floor) went down about half. It may not be enough for Auger but it is inexpensive. TMC, who advertises in most JMSA journals, has a full set of platforms. Phone:508-532-6330, fax:508-531-8682. I have no experience with their service. You wrote:
} We are in need of a vibration isolation pad for an Auger spectrometer, } which has the dimensions of a standard SEM. Can anybody recommend a } manufacturer or can recommend homemade solutions? } Best of luck, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
} Y'all are an encyclopedia of knowledge, and I want to check out a } volume... I have just recently been learning SEM and all the associated } procedures that precede going on the scope. I have had to learn this all } on my own, which is why I'm having this problem. } When I do a cpd on a sample they always come out looking wrinkled. } Am I going wrong in the cpd or is it some other step. I've heard that some } people do a few purges of the system and some do a lot. I've just purged } the ETOH until I see CO2 snowballs & then let it sit for 15 min. & then } repeat the cycle 2 more times. Is this wrong? } Can someone out there give me some pointers? I'm getting pretty } tired of my stuff looking like raisins all the time. = ( } } } Thanks in advance! } } Paula = ) } } Paula Ssicurello } UC Berkeley } Electron Microscope Lab } psic-at-uclink4.berkeley.edu
The main problem particularly newcomers to CPD have is trying to do everything too quickly. I'm no expert at CPD having only done it a few times, but I have been told:
a) when flushing, it need to be done SLOWLY - if the CO2 boils (or even simmers gently), then the specimen will be wrecked.
b) It is important to leave your specimen in the liquid CO2 for long enough for proper infiltration, and that this needs doing at least two or three times to make sure all the acetone/alcohol is displaced - even for small specimens (1mm), this can take 3-4 hours. Larger specimens might require 24 hrs.
c) At the end, when releasing the CO2 gas, again it must be done slowly - 15-20 mins. Sudden pressure releases will blow the specimen apart.
Regards,
--------------------------------------------------------------- Dr L. P. Stoter Technical Editor, MICROSCOPY & ANALYSIS Technesis 17, Rocks Park Road email: LPS-at-teknesis.demon.co.uk Uckfield, E. Sussex Phone: +44 (0)1825 766911 TN22 2AT Fax: +44 (0)1825 766911 United Kingdom ---------------------------------------------------------------
Thanks to all who responded to my question about teaching supplies. I received about 20 responses. They fell into three basic types 1) give each student an allotment at the beginning of the semester and require them to buy more if they use it up; 2) charge each student a flat fee and let them use at will; 3) require them to but their own supplies at a local camera store. Some labs separated EM film (which can be hard to purchase locally) from paper (which is stocked locally).
I'm leaning towards using options #1 or #3.
Thanks again.
Bob
Robert R. Wise Plant Physiologist and Director, UWO Electron Microscope Facility Department of Biology University of Wisconsin Oshkosh Oshkosh, WI 54901 (414) 424-3404 tel (414) 424-1101 fax wise-at-uwosh.edu
You should look into Neurolucida by MicroBrightField. It is a 3D morphometric analysis system. MBF also is also active in stereology analysis and was an exhibitor at the Soc. Neurosci meeting. You can get more info from their web site at www.microbrightfield.com/microb/ or via email at info-at-microbrightfield.com.
On Thu, 27 Feb 1997, Cheri Owen wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } A friend of a friend is looking for a morphometric system with Scope, } stylus, PC etc. They saw some at the recent Neurosci meetings, but didn't } keep the brochures. Any info would be greatly appreciated. } } Thanks } } Cheri Owen } Detroit Neurotrauma Institute } Wayne State University } Detroit, MI } } }
Edmund Glaser, D. Eng. Dept. Physiol. Univ. Md. School. Med. Baltimore, MD 21201 USA Ph: (410) 706-5041 Fax: (410) 706-8341
We are trying to photograph bacteria using phase contrast optics. We are having trouble getting a thin enough layer to obtain 'fully focused" micrographs at 100x under oil. We are using a fixed suspension mixed with "aquatex" mounting medium. Any suggestions? Thanks David Wild
I am sorry that this is a little late. I will be trying to get this message out to the individual authors who will be presenting who do not receive it through the Listserver. If you are the principal author or a coauthor on a paper being presented at Symposium Z at the Spring '97 MRS meeting, please let me know that you received this anouncement ASAP.
The papers that you are submitting to the "Workshop on TEM Specimen Preparation-IV", Symposium Z should be tutorial in nature. They should be written in a "how I did it and here's how you can do it" mode with all the tidbits and tricks-of-the-trade that make your technique work. Follow the authors guidline for the paper for the MRS proceedings except for length. Use as many pages as you need to tell your story. Use as many pictures and diagrams that you need to get the information across. If you have any questions, contact me or Ron Anderson.
- -Scott Walck
Ron Anderson, Chair IBM, ZIP E-70 East Fishkill Facility Hopewell Jct., NY 12533 *(914)892-2225 (-2003 FAX) ron-anderson-at-vnet.ibm.com
Scott D. Walck, Co-Chair Materials Directorate, Wright Laboratory Wright Patterson AFB, OH 45433-7750 *(513)255-5791 (-2176 FAX) walcksd-at-ml.wpafb.af.mil
Can anyone please tell me the name of staining books in the field of Immuno-bed tissues as well as the companies from which I can order it. We are interested in - Histology and Pathology - Immunofluorescence - Immunohistochemistry - Histofluorescence - Enzymehistochemistry
Thanks Hildagonda van der Merwe
University of Pretoria Faculty of Veterinary Science Dept. of Pathology Onderstepoort 0110
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello All, } } } Y'all are an encyclopedia of knowledge, and I want to check out a } volume... I have just recently been learning SEM and all the associated } procedures that precede going on the scope. I have had to learn this all } on my own, which is why I'm having this problem. } When I do a cpd on a sample they always come out looking wrinkled. } Am I going wrong in the cpd or is it some other step. I've heard that some } people do a few purges of the system and some do a lot. I've just purged } the ETOH until I see CO2 snowballs & then let it sit for 15 min. & then } repeat the cycle 2 more times. Is this wrong? } Can someone out there give me some pointers? I'm getting pretty } tired of my stuff looking like raisins all the time. = ( } } } Thanks in advance! } } Paula = ) } } Paula Ssicurello } UC Berkeley } Electron Microscope Lab } psic-at-uclink4.berkeley.edu
Paula, It sounds like your samples are still "wet". Make sure that your are dehydrating your samples long enough. The ETOH or acetone must be 100% pure. The time you are alloting in the CPD is too short. This will also leave your sample wet with acetone or ETOH. When you drain the ETOH do it slowly and place a dry Kimwipe close and infront of the vent. When it no longer gets wet after a few purges the ETOH is gone. Then Let it sit for 30 min. more. Bring the temp. of the chamber up to about 37C slowly. Vent the CO2 over 10 min until the pressure is zero. The above is all sample dependent and must be worked out by trial and error. Plant material will take longer than a cell cultuer on a cover slip.
Greg Rudomen University Microscopy Imaging Center S.U.N.Y. Stony Brook Greg-at-umic.sunysb.edu
This is to remind everyone that extended abstracts for Microscopy & Microanalysis '97 need to be subimtted by MARCH 15! That is this Saturday! Papers received after that date will not be accepted.
If you have misplaced or have not received a bulletin or need more information, you can contact
Microscopy & Microanalysis '97 4 Barlows Landing Rd., Suite 8 Pocasset, MA 02559 phone: 508-563-1155 Toll-free: 800-538-3672 FAX: 508-563-1211
Email: BusinessOffice-at-MSA.Microscopy.com or WWW: http://www.MSA.Microscopy.com
Meeting information is also available at http://www.bright.net/~strecker/msno/mm97.html
Poster instructions can be found at the MRS web site, www.mrs.org
You should have recieved author instructions. Please contact MRS if you have not. The Materials Research Society, 9800 McKnight Road, Pittsburgh, PA 15237-6006 Phone: 412/367-3004, Fax: 412/367-4373, Email: info-at-mrs.org
If you haven't, this should get you started.
For camera-ready text, on an 8-1/2 x 11 inch paper, the margins are left: 1 inches right 1 inches top: 1/2 inch bottom: 1 inch Page number at 1/2 inch from bottom of page.
That gives 6.5 inch wide x 10 inch box to put your text, single spaced, 12 point font Times Roman. Use the AIP reference style or the MRS refernce style. The manuscripts will be reduced by 26%. Scale markers on all micrographs. Check out other proceedings for style if you haven't received your instructions.
Bring the original manuscript and (3) photocopies to the meeting. You will need to have the MRS copyright form filled out. Check the bulletin boards on where to take your paper. Ron or myself will be collecting them from you and will post an announcement.
The Royal Microscopical Society Computers in Microscopy ,September 1997, Cambridge CALL FOR PAPERS
A Colloquium on various aspects of the application of computers to microscopes and microscopical techniques is being planned by the Royal Microscopical Society, to be held in the University Engineering Department, Trumpington Street, Cambridge, on Thursday 25th September 1997.
The colloquium is intended to cover recent progress made in the use of computers for the assessment, interpretation, restoration and enhancement of microscopical images, including, but not restricted to applications in optical and all kinds of electron microscopy. Of particular interest will be contributions concerning new approaches and techniques including 3D measurements and profiling, wavelets and fractals. It is hoped that papers will cover some of the comparatively new fields of application, including multi-channel imaging, instrumental control and remote microscopy. It is proposed also to organise a small exhibition of products and materials by local organisations involved in this area, as well as a visit to laboratories in the University engaged in this kind of work.
The colloquium takes place immediately after a three-day course, also entitled 'Computers in Microscopy', and it is intended that the two events be complementary. ( Please note that it is not necessary to attend the course in order to register for the colloquium.)
If you would like further information about the Colloquium and/or the Course please contact Rebecca Morden either by email: info-at-rms.org.uk, by telephone: +44 (0)1865 248768 or by fax: +44 (0)1865 791237.
Prospective contributors are invited to submit a synopsis of approximately 150-200 words, before 30 April 1997, to Dr D M Holburn, University Engineering Department, Trumpington Street, Cambridge CB2 1PZ, U.K. Enquiries may be made by electronic mail, to dmh-at-eng.cam.ac.uk, by facsimile to +44 1223 332662, or by telephone to +44 1223 332775.
******************************************************************************* Rebecca Morden Course Organiser Royal Microscopical Society, 37/38 St Clements, Oxford, OX4 1AJ, UK. Tel (0)1865 248768, Fax (0)1865 791237, email info-at-rms.org.uk *******************************************************************************
Location : Carolinas HealthCare System Charlotte, North Carolina, USA
Shift: Full-time, M-F, 8a-4:30p.
Essentials: Fix, dehydrate, and embed biological tissue specimen for transmission e.m. Thick and thin sectioning, Basic operation of Philips CM10 electron microscope. Develop and print photomicrographs. Maintain logs and records of work,
Requirements: BA/BS degree in related area. Minimum 1.5 year relatively recent experience in electron microscopy, preferably in a work setting. Good "people skills."
} We are in need of a vibration isolation pad for an Auger spectrometer, } which has the dimensions of a standard SEM. Can anybody recommend a } manufacturer or can recommend homemade solutions?
There are some low cost pneumatic mounts available from Barry Controls in a wide range of load handling capacities. The trade name is Stabl-Levl. I used these to successfully isolate a heavy FTIR unit that was sensitive to vibration. Unless you can attach them directly to your instrument, however, you will still need a stiff plate to set it on with the pneumatic mounts between the plate and the floor. The address and phone # I have for Barry Controls is: 700 Pleasant Street Watertown, Mass. 02172 617-923-1150 Haven't ordered from them in a few years, so you might need to check it if you can't connect. Disclaimer: I have no interest in Barry Controls other than being a satisfied customer.
Andy Buechele The Catholic University of America 409 Hannan Hall Washington, D.C. 20064 (202) 319-4995 FAX: (202) 319-4469
Our Auger with scanning tunnel microscope (with console)hangs from ceiling beams on three springs if you need details you may contact Prof. Achete (achete-at-metalmat.ufrj.br) in this department Best regards Prof.Walter A.Mannheimer Metallurgy and Materials Engineering Federal University of Rio de Janeiro POBox 68505 21945 Rio de Janeiro, Brazil Voice +5521 280-7443 (Dept.office) +5521 590-0579 (direct) Fax +5521 290-6626 Email: wamann-at-metalmat.ufrj.br
Can anyone tell me the best way to mount a sample for hot stage work. We have a cross section of a silicon nitride sample that we want to examine at elevated temperatures (up to about 1000 deg C, if our hot stage will get us there!) The sample is a cross section made by tripod polishing and it must be mounted on a Mo or Ta washer for insertion into the scope, how are we going to "glue" it to the ring so that the "glue" wont decay and the sample fall off or move about at high temp? What we need is a very low vapor pressure adhesive that can withstand high temperatures!
Any help appreciated. Thanks.
Jfm.
John Mansfield North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (313) 936-3352 FAX (313) 936-3352 Cellular Phone: (313) 715-2510 (Leaving a phone message at 936-3352 is preferable to 715-2510) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html
Due to a projected streamlining of the review and production process, abstracts for the June meeting of the Microscopy Society of Canada can be accepted up to 2 weeks later than previously advertised, i.e. they must be received in Edmonton by 1 April.
The deadline for conference pre-registration is 1 May 1997, as generally advertised (but NOT 1 March as stated on the registration form!)
Ray Egerton, Physics Dept, University of Alberta, Edmonton, Canada T6G 2J1 Phone: 403-492-5095, FAX: 403-492-0714, e-mail: egerton-at-phys.ualberta.ca ------------------------------------------------------------------------
Afew weeks ago, there was someone who was doing a survey of the best types of gloves to use in an EM lab. Of course I didn't save the information passed on, and of course now we're interested in those results here in our lab. If anyone has those results, please let me know. Also, we work with a lot of the low-temperature resins (Lowicryls, LR Gold), if anyone knows of the best gloves to work with that stuff let me know that too.
Thanks oodles!
Paula = )
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
I would like tofind someone in the S.F. Bay area that has a freeze-fracture machine that would be available for doing samples. I have a colleague in the area who is looking for a facility where she might be able to do some samples. We would be very grateful for any leads. Thanks in advance. ML
Mei Lie Wong Department of Biochemistry HHMI-UCSF Ph. 415-476-4441 Fax 415-476-1902 email wong-at-msg.ucsf.edu
I am about to replace the sputter ion pump elements in our two JEOL TEM's. JEOL used to be able to recondition the elements used in their 2000FX microscopes but no longer provide this service. Can anyone suggest another company that does this sort of work? How long can I expect a reconditioned unit to last compared with a new one and what sort of cost would be reasonable given that JEOL are charging $2.5k and $4.5k (Australian dollars) for new units? Thanks in advance for any advice you can give.
Mark Blackford TEM Group Materials Division, ANSTO PMB 1, Menai, N.S.W. Australia 2234 Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
The Oklahoma Microscopy Society (OMS) will be hosting its 19th Annual Spring Workshop on Friday, April 4, 1997.
Topic: FORENSIC MICROSCOPY
MORNING SESSION: 129 George Cross Hall, University of Oklahoma in Norman, OK.
8 - 9 am: Registration
9 - 9:05: Greetings
9:05 - 9:50: Guest speaker: Mark Betts (from Oxford Instruments) will speak on "Microanalysis in the Forensic Laboratory"
9:50 - 10: Break
10 - 12: Guest speaker: Keith Ferrell (from the Oklahoma State Bureau of Investigation (OSBI)) will speak on "Overview of Forensic Microscopy"
12 - 1:30pm: Lunch/Executive Meeting/Open Meeting
AFTERNOON SESSION: OSBI Central Laboratory, 2132 N.E. 36th St. in Oklahoma City
1:30 - 2: Travel to the OSBI
2 - on: Tours of the OSBI Facility
REGISTRATION (includes lunch):
You can download a copy of the registration form by visiting the OMS website at http://www.ou.edu/research/electron/oms/ maintained by Dr. Scott Russell, Nobel Electron Microscopy Lab at the University of Oklahoma.
OMS members: the Newsletter was sent out Monday, March 10, 1997 so you should be receiving it this week. A registration form/envelope has been included in the newsletter. Please return it as soon as possible so as to estimate numbers for lunch and parking permits.
Student and Professional Members: $5 Student Non-Members: $10 Professional Non-Members: $15
Pre-register until April 2, 1997 with Phoebe Doss OMS President Elect Dept. of Plant Pathology 110 NRC Oklahoma State University Stillwater, OK 74078 (405) 744-7995 Email: pjdoss-at-okway.okstate.edu
Your question, "how to stain enzymes for EM" is so broad as to be unanswerable, I think. Which enzymes?, Plant tissue or animal? Cells grown in culture? I am sure someone on the list can help if your question is more specific.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (908)-235-4583; fax -4029 e-mail: mcauliff-at-.umdnj.edu ***************************************************************
On 03/10/97 17:21:17 you wrote: } I would like tofind someone in the S.F. Bay area that has a freeze-fracture } machine that would be available for doing samples. I have a colleague in } the area who is looking for a facility where she might be able to do some } samples. } We would be very grateful for any leads. Thanks in advance. ML } } Mei Lie Wong } Department of Biochemistry } HHMI-UCSF } Ph. 415-476-4441 Fax 415-476-1902 } email wong-at-msg.ucsf.edu
Dear Mei, Please call Nancy Smith at Cal State Hayward (main number 510-885-3000). I am sure she cal help you. Regards
Igor C. Ivanov, SIMS,Auger,TEM,SEM Senior Scientist Analytical Services RJ LeeGroup, Inc. Contract Research 530 McCormick Str. (510)-567-0480 phone San Leandro, CA 94577 (510)-567-0488 FAX
} Return-Path: {das3-at-Lehigh.EDU} } X-Sender: das3-at-mail.Lehigh.EDU } Date: Tue, 11 Mar 1997 16:26:22 +0100 } To: Donna Jacobs {jacobs-at-uimrl7.mrl.uiuc.edu} } From: das3-at-Lehigh.EDU (David A. Smith) } Subject: Re: Open Position - Research Electron Microscopist } } DR. DAVID SMITH PASSED AWAY SEPTEMBER 1996. PLEASE REMOVE FROM ALL E-MAIL } AND MAIL LISTINGS. THANK YOU. } } MAXINE MATTIE } } David A. Smith, Department of Materials Science and Engineering, Lehigh } University, 5. E. Packer Ave., Bethelehem, PA 18015, Ph 610 758 4231, Fax } 610 758 4244 } } } Donna Jacobs MRL Administration University of Illinois 104 South Goodwin Avenue Urbana, Illinois 61801 Phone (217) 244-2944 Fax (217) 244-2946 email - jacobs-at-uimrl7.mrl.uiuc.edu
Good Afternoon (Central Std Time USA) all Microscopists:
I have just had a request to determine the refractive index of zinc pyrithione, a crystalline, colorless, transparent powder. Does anyone know what the RI is? It's not in McCrone Atlas. I will be getting a sample Monday and with a set of Cargille RI liquids, I will determine the RI.
Alternatively, I have a B & L Abbe type refractometer used to determine refractive index of liquids. However, I thought I read somewhere that it could be used to determine the RI of solids. If this is in fact possible, does anyone out there know how?
Thanks to all who responded to my cpd inquiry. The consensus seems to be that fresh, dry 100% ETOH is very important and that I need to keep my samples longer in the liquid CO2 to get a better exchange with the ETOH.
I'll try out all the suggestions. Hopefully, the only raisins I'll get from now on will be in my breakfast cereal. = )
Thanks again.
Paula = )
p.s. I'm going to phone some vendors of the resins & such to see if they've done studies as to which type of gloves work best. I'll keep you posted.
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
I am acquiring an ISI SX-30E and need a copy of the manual for this instrument without paying the hundreds of dollars Topcon wants for such a manual. Any suggestions where I might be able to get a xerox of the manual or borrow a copy to xerox. Any help would be appreciated.
Duniway Stockroom Corp. (800-446-8811; info-at-duniway.com) advertise a service for rebuilding sputter ion pumps (p. 25 of their catalog), and state that a properly rebuilt pump should "have the same performance as a new pump". The cost of rebuilding varies from about $400 to about $3000, depending on the size and type of pump No commercial interest - just trying to be helpful.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
Does anyone who performs vascular casting know where to obtain a product called Mercox CL2? A fellow electron microscopist has heard that it is the latest thing in vascular casting when mixed with methyl methacrylate but hasn't been able to get any further info. If anyone knows who makes it and where it can be bought we would like to hear.
Thanks in advance
Richard Easingwood South Campus Electron Microscope Unit School of Medical Sciences University of Otago PO Box 913 Dunedin NEW ZEALAND
For the theory you need to look up Snell's Law and Molecular refractivity. The RI relates to the velocity of light through various substances. The difference between air and water RI results in a stick appearing angled at the interface. Transparent specimen become invisible if immersed in a liquid of identical RI. This is extensively used in police scientific work were glass fragments can be identified to come from a crime scene. Very simple, put a sample onto a microscope slide. Add a drop of RI solution. If the submersed specimen becomes invisible under the microscope, than the refractive index of the specimen is that of the RI test solutions. Some experimenting to find the right solution is required. Good luck. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 300+ Links, MSDS ************************ http://www.proscitech.com.au ---------------------------------------------------------------------------- ----- } From: Damian_Neuberger_at_RLT013-at-ccmailgw.mcgawpark.baxter.com } To: microscopy-at-Sparc5.Microscopy.Com } Subject: Refractive Index } Date: Wednesday, 12 March 1997 6:49 } } I have just had a request to determine the refractive index of zinc } pyrithione, a crystalline, colorless, transparent powder. Does anyone } know what the RI is? It's not in McCrone Atlas. I will be getting a } sample Monday and with a set of Cargille RI liquids, I will determine } the RI.
} Transparent specimen become invisible if immersed in a liquid of identical } RI. This is extensively used in police scientific work were glass fragments } can be identified to come from a crime scene. Very simple, put a sample } onto a microscope slide. Add a drop of RI solution. If the submersed } specimen becomes invisible under the microscope, than the refractive index } of the specimen is that of the RI test solutions.
Unfortunately it is not quite that simple. If your material is a glass and/or has one refractive index this procedure will work fine. Don't forget to correct for temperature and wavelength!
But-- If your sample has two or more refractive indexes the procedure becomes a lot more complicated. You need to separate one refractive index from another and this can be done with the polarized light microscope with a lot of practice. I suggest "Handbook of Chemical Microscopy, Vol One" by Chamot and Mason, if you can find a copy, as a reference. The procedures and techniques are too detailed for a short note.
Best wishes.... Frank
PS i was unable to find any optical data in Winchell's "The Optical Properties of Organic Compounds"
It is possible to determine the RI of SOME solids with the B&L Abbe refractometer. But I don't know if it is possible to do this on a powder; I have never done it. The more critical requirements for doing a solid (if you are still interested) are:
1) You must have very intimate contact between the solid and the lower prism. This is normally accomplished by polishing the sample such that it is extremely flat, and then a contact fluid (1-bromonapthalene) is used to form the intimate contact. Use as little contact fluid as necessary (usually a very small drop).
2) It is important to have a 90 degree angle between the bottom prism and one edge of your sample. This edge also needs to be highly polished, like the contact surface.
Good luck with the powder; if anyone suggests a way to do this with the refractometer, I would also be interested.
Regards,
-Bob ************************************ Bob Citron Chiron Vision 555 W. Arrow Hwy Claremont, CA 91711 ph: (909) 399-1311 email: Bob_Citron-at-cc.chiron.com ************************************
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Good Afternoon (Central Std Time USA) all Microscopists:
I have just had a request to determine the refractive index of zinc pyrithione, a crystalline, colorless, transparent powder. Does anyone know what the RI is? It's not in McCrone Atlas. I will be getting a sample Monday and with a set of Cargille RI liquids, I will determine the RI.
Alternatively, I have a B & L Abbe type refractometer used to determine refractive index of liquids. However, I thought I read somewhere that it could be used to determine the RI of solids. If this is in fact possible, does anyone out there know how?
Osmolarity is an easily measurable characteristic, by e.g. freezing point depression. In simple terms it is the total concentration of solutes (including ions) in a solution, and this does not depend on whether the solutes can cross cell membranes or not. Tonicity relates to the osmotic GRADIENT due to solutes that affects a semi-permeable membrane ( i.e. a membrane that is permeable to water); ONLY solutes that do not cross the membrane contribute to this effect. The two characteristics are different and obviously have vastly different consequences on membranes. Weak bases, even though ionized, have some measurable permeability and so their contribution to tonicity must be much less than their contribution to osmolarity. Osmium is hydrophobic and quite permeable through membranes, so contributes *nothing* to an osmotic gradient that can disrupt the membranes (even assuming it doesn't alter their permeability to tonic agents like sucrose). So osmium contributes nothing to tonicity, but certainly does contribute to osmolarity.
I personally wonder how to express the tonicity of compounds that are somewhat permeable, such as weak bases or short carboxilic acids & aldehydes. I guess the concept of tonicity either doesn't apply or is operational in such cases, depending on the time scale of interest. Can anyone comment or give a reference to a good detailed textbook? Something about reflection coefficients?
Richard
} } } Gary Dietrich Chinga {garyc-at-stud.ntnu.no} 03/12/97 09:31am } } } Hi!
Can someone explain the relation between the tonicity and osmolarity of a fixation solution?
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Gary,
Tonicity is a relative, unitless comparison of one solution to another in which the first solution (presumably the water in your tissue sample) is hypertonic, hypotonic or isotonic to the second solution (presumably your fixative). Osmolarity is an absolute scale (usually in some type of pressure unit) which describes the concentration of osmolytes in a single solution. So you need to know the osmolarity of your tissue so you can set the osmolarity of your fixative to be isotonic with respect to the tissue.
Cheers
Bob
Robert R. Wise Plant Physiologist and Director, UWO Electron Microscope Facility Department of Biology University of Wisconsin Oshkosh Oshkosh, WI 54901 (414) 424-3404 tel (414) 424-1101 fax wise-at-uwosh.edu
I have a user trying to follow a new protocol which uses "an antistatic solution of ... Denkil-SEM (Hodogaya Chemical)" but I have no idea what "Denkil-SEM" is can anyone out there give us a hint? (Before we start trying other antistatic solutions, i.e. Static Guard, or Cling Free!).
Thanks.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513-529-5712 Fax: 513-529-4243 E-mail: edelmare-at-muohio.edu
I have a question which has undoubtedly been answered before...! We are going to buy a flat bed scanner, probably a Microtek with a transparency adapter. We would primarily use it for scanning in TEM and SEM (polaroid) negatives. Should we go for the 24-bit color, 300 x 600 dpi version, or the more expensive (ouch) 30 bit color, 600 x 1200 dpi version? Our resident computer guru says that we should go with the cheaper version because we are unlikely to have an output source with better than 600 dpi for most stuff. It's true; the printers around here are mostly 300 - 600 dpi. Is there a compelling reason for the better model? Will I be really sorry in a year if I don't?
Thanks you all for all the expertise and advice on so many different topics!
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
I've been checking around on the glove issue. It seems that there is no one glove that works for everything (darn!). Usually if it's good for one thing, it's lousy for another. We work with Lowicryl resins a lot and the best glove to use is........
Nitrile, those blue smelly things that don't stretch worth a darn.
I also received some info. from a paper (thanks, Bill = ) ), that told of an actual test done with various resins & such on different makes of gloves. The paper is "Glove Material For Handling Epoxy Resins" by David L. Ringo, Douglas R. Read and Eugene H. Cote-Robles in the Journal of Electron Microscopy Techniques 1:417-418 (1984). Pretty interesting stuff!
Safety in the lab is an important thing. So each lab must make it's own mind as to which type of glove they want to use. The important thing is that you get your students to glove up and work in the fume hoods and be aware that almost all of the chemicals in an EM lab are hazardous.
Happy Scoping!
Paula = )
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
********************************************* * Metropolitan Microscopy Society Meeting * *********************************************
Date: Wednesday, March 26, 1997
Time: 10:00 AM
Place: Howard Johnson Lodge, 393 Route 17, Paramus, NJ
Directions: The Howard Johnson Lodge is on the southbound lane of Route 17. From the Garden State Parkway, use Exit 163 if you are northbound, and Exit 165 if you are southbound.
10:00 - 10:30 am Registration ($5.00), Coffee and Danish.
10:30 - 10:45 am Introductory remarks and society announcements -- Philip Flaitz.
10:45 - 11:30 am HIGH RESOLUTION LOW VOLTAGE SCANNING ELECTRON MICROSCOPY, Dr. Frederic Cosandey, Dept. of Ceramic and Materials Science, Rutgers, The State University of New Jersey, Piscataway, NJ.
Scanning Electron Microscopy at low voltages (0.2 - 5 keV) has many advantages over more conventional electron energies such as reduced electron range, increased secondary electron yield, reduced radiation damage and charging artifacts. However, in order to maintain high resolution at the lowest voltages, special lens designs are required to correct for the chromatic aberration of the probe forming objective lens. In addition, improved secondary electron detection systems must be implemented. In this talk, the relative merits of various objective lens designs will be discussed with special emphasis on the electrostatic lens with retarding field concept. Also, the unique advantages of low voltage SEM for secondary and backscattered electron imaging modes will be discussed. A few specific examples will be presented including domain size measurements in block co-polymers, oxide surface structure determination and phase identification in nanocomposites.
11:30 - 12:15 pm MICROBEAM ANALYSIS OF POLYMERS, DRUGS AND INOR- GANIC MOLECULES BY INFRARED MICROSPECTROSCOPY, Dr. John A. Reffner, Spectra-Tech Inc., Shelton, CT 06484
Infrared microspectroscopy (IMS) is a growing technology for microbeam analysis of organic and molecular materials. As the fusion of scanning electron microscopy with x-ray emission spectroscopy created an exciting way for microscopists to investigate elemental composition, so has IMS impacted the way we now investigate the molecular chemistry of materials. IMS is essentially a photon probe --- producing spectral data by the absorption or reflection of infrared radiation. Applications to polymers, pharmaceuticals, electronics and forensics illustrate the wide range of uses of IMS. In reviewing IMS technology, both the strengths and limitations of current instruments are presented with a look at directions for future developments.
12:30 - 1:30 pm Lunch.
For additional information, please contact:
Phil Flaitz, IBM Analytical Services pflaitz-at-vnet.ibm.com, 914-892-3094
I know this question has been asked here before, so bear with me. Where can I get one of those red antistatic guns, now that they are no longer handled by Fullam? Their representative told me that there is supposedly a hefty supply of these units still in the U.K., and that Fullam doesn't handle them anymore because they'd have to buy more than they could sell. I know Discwasher used to distribute them. Does anyone know of the English company that has this huge supply? Are there any distributors? Is there any similar product? Appreciate any leads.
} Can someone explain the relation between the tonicity and osmolarity } of a fixation solution?
= = = = = = = = = = = = = = = =
Osmolarity is a measure of one of the colligative properties (osmotic pressure) of a solution. In a rough sense it is the measure of the amount of solute in a solution, but the degree of dissociation of the solute affects its osmolarity, so osmolarity is not necessarily a direct measure of molar concentration. (For example, a 1 molar solution of NaCl will have *approximately* twice the osmotic pressure [osmolarity] of a 1 molar solution of sucrose, since the NaCl dissociates into 1 molar Na+ and 1 molar Cl-.)
Two solutions (not necessarily of the same solute or of a single solute) are isosmotic if they have the same osmolarity (osmotic pressure). If two isosmotic solutions are placed on opposite sides of a semipermeable membrane, the osmotic pressure on each side is the same and there is no *net* movement of water across the membrane. However, if the osmolarity of the two solutions is not the same, then water will move across the membrane from the hypo-osmotic solution (more dilute) to the hyperosmotic solution (less dilute) until the osmotic pressure on each side is equal (a number of other factors affect this as well, but let's ignore them here).
Tonicity refers to the response of *cells or tissues* to the solutions in which they are immersed. If cells are placed in a hypertonic solution, net movement of water will be out of the cell, causing the cell to shrivel. If cells are placed in a hypotonic solution, net movement of water will be into the cell, causing the cell to swell or burst. Tonicity is useful only in reference to a particular cell or tissue.
Thus, the microscopist wishes to add sufficient solute(s) to the fixation solution so that the solution has the correct osmolarity (measured in milliosmols) so that it will have the desired tonicity with respect to the cells that are being exposed to the fixative. [There is debate whether the solution should be slightly hypertonic or hypotonic, but then that is another subject about which we may choose to debate.]
To summarize, osmolarity is a measurement of solute concentration (measurement can be made in a beaker). Tonicity is a comparison of osmolarities between a cell and the solution to which it is exposed.
I hope that this does not leave readers more confused than they were before. }
______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 The College of New Jersey fax: (609) 771-2674 Trenton, NJ 08650-4700
The 30 bits will give you additional shadow and highlight detail that 24 bits will not provide. Now, you may ask 'my software can only handle 24 bits so what good does 30 bits do when I will have to convert it down?'. When you convert the file to 24 bits, the system will choose from the best 24 bits to produce the converted file. I would equate it to a survey or poll - the larger your sample, the more accurate your results.
John D. Warren Area Sales Manager Digital Products Polaroid Corporation
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have a question which has undoubtedly been answered before...! We are going to buy a flat bed scanner, probably a Microtek with a transparency adapter. We would primarily use it for scanning in TEM and SEM (polaroid) negatives. Should we go for the 24-bit color, 300 x 600 dpi version, or the more expensive (ouch) 30 bit color, 600 x 1200 dpi version? Our resident computer guru says that we should go with the cheaper version because we are unlikely to have an output source with better than 600 dpi for most stuff. It's true; the printers around here are mostly 300 - 600 dpi. Is there a compelling reason for the better model? Will I be really sorry in a year if I don't?
Thanks you all for all the expertise and advice on so many different topics!
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
I would suggest to check out the speed as well as the resolution. We have an old HP-IIcx here that easily beats a newer cheaper scanner hands down. The software is also not as convenient as HP's deskscan package.
Regarding resolution, remember it will take a few of those printer pixels to dither up the gray scale for each of your digitized pixels. Thus, a 600 dpi printer may only be able to handle about a 200 dpi image. Of course if you are scanning 35 mm slides and enlarging them, you will need the high res scanning.
At 11:26 AM 3/12/97 -1000, Tina wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Donald Lovett's explanation quoted below is very good except that it perpetuates the myth that tonicity is related to the difference between two OSMOLARITIES on opposite sides of the membrane. That is ONLY true if the membrane involved is impermeable to ALL of the solutes contributing to the osmolarity. Since formaldehyde is soluble in both benzene and chloroform I expect it to be quite permeable across cell membranes. Thus it will contribute to osmolarity, but should contribute little to tonicity. The tonicity of your fixative should be adjusted with other components, and the final osmolarity of an isotonic fixative will be approx 300 mOsm PLUS the value of osmolarity contributed by the formaldehyde present.
To reiterate, IF a compound is permeable across the membrane it contributes to osmolarity but does not contribute to the osmotic GRADIENT. Thus it does NOT contribute to tonicity.
Caveats: Practically speaking, the rate at which a permeable solute crosses the membrane will affect the transient forces on the membrane. I assume tonicity is ill-defined unless the system is at equilibrium. The situation becomes even more complicated if the compounds added (such as aldehydes or alcohols) alter the permeability of the membrane for other buffer components present. And if differences in rates of diffusion through a chunk of tissue are involved, are we really still talking about tonicity?? That calls for empirical, not theoretical, optimization of the recipe used !!
On that note, how many of you microscopists have actually compared e.g. confocal images of cell cultures fixed with buffers of different tonicity with equivalent images of live cultured cells? The time to diffuse through a monolayer of cells should be negligible. What do you find is optimum?
Richard
} } } Donald Lovett {lovett-at-tcnj.edu} 03/12/97 05:03pm } } } Osmolarity is a measure of one of the colligative properties (osmotic pressure) of a solution. In a rough sense it is the measure of the amount of solute in a solution, but the degree of dissociation of the solute affects its osmolarity, so osmolarity is not necessarily a direct measure of molar concentration. (For example, a 1 molar solution of NaCl will have *approximately* twice the osmotic pressure [osmolarity] of a 1 molar solution of sucrose, since the NaCl dissociates into 1 molar Na+ and 1 molar Cl-.)
Two solutions (not necessarily of the same solute or of a single solute) are isosmotic if they have the same osmolarity (osmotic pressure). If two isosmotic solutions are placed on opposite sides of a semipermeable membrane, the osmotic pressure on each side is the same and there is no *net* movement of water across the membrane. However, if the osmolarity of the two solutions is not the same, then water will move across the membrane from the hypo-osmotic solution (more dilute) to the hyperosmotic solution (less dilute) until the osmotic pressure on each side is equal (a number of other factors affect this as well, but let's ignore them here).
Tonicity refers to the response of *cells or tissues* to the solutions in which they are immersed. If cells are placed in a hypertonic solution, net movement of water will be out of the cell, causing the cell to shrivel. If cells are placed in a hypotonic solution, net movement of water will be into the cell, causing the cell to swell or burst. Tonicity is useful only in reference to a particular cell or tissue.
Thus, the microscopist wishes to add sufficient solute(s) to the fixation solution so that the solution has the correct osmolarity (measured in milliosmols) so that it will have the desired tonicity with respect to the cells that are being exposed to the fixative. [There is debate whether the solution should be slightly hypertonic or hypotonic, but then that is another subject about which we may choose to debate.]
To summarize, osmolarity is a measurement of solute concentration (measurement can be made in a beaker). Tonicity is a comparison of osmolarities between a cell and the solution to which it is exposed.
I hope that this does not leave readers more confused than they were before. }
______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 The College of New Jersey fax: (609) 771-2674 Trenton, NJ 08650-4700
I'd like to forward this announcment of an open post-doc position to the list:
} The Laboratoire d'Analyse des Materiaux (LAM), of the "Centre de Recherche } Public-Centre Universitaire" in Luxembourg, has an immediate opening for a } two years post-doc position. } The core of the subject to be covered deals with analytical TEM (EELS and } EDX) : sensitivity, quantification, artifacts,... on complex samples. } As a side subject, this person will have to help in optimizing the } preparation of cross sectional samples: ion dimpling/microtome sectioning. } } If you have the necessary qualifications, please send your resume to : } Dr. H.N. Migeon } Director of LAM } 162a, Avenue de la Faiencerie } L-1511 Luxembourg } After a first selection, interviews will take place in Luxembourg. No } funding will be provided for overseas trips. } } Henri-Noel Migeon
Best regards,
Petra
-------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public Centre Universitaire (CRP-CU) Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpcu.lu or 100112.2335-at-compuserve.com
On Wed, 12 Mar 1997 WARRENJ1-at-cliffy.polaroid.com wrote:
} The 30 bits will give you additional shadow and highlight detail that } 24 bits will not provide. Now, you may ask 'my software can only } handle 24 bits so what good does 30 bits do when I will have to } convert it down?'. When you convert the file to 24 bits, the system } will choose from the best 24 bits to produce the converted file. I } would equate it to a survey or poll - the larger your sample, the more } accurate your results.
This statement seems a little bit tricky. 24 color bits gives one 256 grey shadows. If one want only to store and reproduce images - this is more than enough (128 is O'K for viewing). If one want to quantify grey levels for processing, 256 levels is also usually enough because of narrow linear range of film, grain, etc. Interpolation will give better results. If one want to waste money - I have no reasons not to this :)
Need to find a floor model lab vacuum coating systemwith : Defussion pump, HV(2000 Volt) 200 ma. feed thru, low voltage high current feed thru, cryotrap,etc. Also looking for an older Balzers 360 Freeze Fracture system(any condition) and an old hummer or a sputtering head for Au-Pd. By the way does anyone keep up with the "Particle-Protein" Freeze Fracture theories and can you send me some references? Been a while.
Jeff Day 3208 Statler Mesquite, Texas 75150 Day Phone:972-975-4338
Hello all, I have been looking at very thin (~10-30 A) multilayers of silicon oxide and nitride. I have very simplistically assumed that the position of the interface lies half-way between the first bright and dark fringe on each side of the interface. Does anyone know whether this is a valid assumption - and if there's any software that I can use to simulate the image? (Or, failing this, where I can get hold of the theory to let me do this myself).
Many thanks in advance,
Richard Beanland, GMMT Ltd., Caswell, Towcester, Northants NN12 8EQ UK
Tina Carvalho wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I have a question which has undoubtedly been answered before...! We are } going to buy a flat bed scanner, probably a Microtek with a transparency } adapter. We would primarily use it for scanning in TEM and SEM (polaroid) } negatives. Should we go for the 24-bit color, 300 x 600 dpi version, or } the more expensive (ouch) 30 bit color, 600 x 1200 dpi version? Our } resident computer guru says that we should go with the cheaper version } because we are unlikely to have an output source with better than 600 dpi } for most stuff. It's true; the printers around here are mostly 300 - 600 } dpi. Is there a compelling reason for the better model? Will I be really } sorry in a year if I don't? } } Thanks you all for all the expertise and advice on so many different } topics! } } Aloha, } Tina } } http://www.pbrc.hawaii.edu/bemf/microangela } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } **************************************************************************** Tina - buy the most true optical resolution and pixel depth you can afford - not so much for the SEM stuff but for the TEM negatives. The scanner should have enough optical (not interpolated) resolution to over sample the film resolution by at least a factor of 2.5 to 3 (the Nyquist sampling limit you know) otherwise you will not be able to treat the digitized images like the original negative - i.e. information will be lost! The company that makes tha scanner probably also makes a 1k x 2k version which would be even better.
} } Donald Lovett's explanation quoted below is very good except that it } perpetuates the myth that tonicity is related to the difference between } two OSMOLARITIES on opposite sides of the membrane. That is ONLY } true if the membrane involved is impermeable to ALL of the solutes } contributing to the osmolarity. ......
} Richard:
Thanks for your extended clarification of the subject. I agree entirely with your comments. I had tried to focus on the difference between the two terms and simplify my response with the caveat: } } "(a number of other factors affect this as well, but let's ignore them here)." } I also agree that the response of the cell to the solution (as evaluated by trial and error) is the most important aspect to microscopists, irrespective of how one names or measures the composition of the solution.
Don } } ______________________________________________________________________ } Donald L. Lovett e-mail: lovett-at-tcnj.edu } Assoc. Professor, Dept. of Biology voice: (609) 771-2876 } The College of New Jersey fax: (609) 771-2674 } Trenton, NJ 08650-4700 } } } } } }
______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 The College of New Jersey fax: (609) 771-2674 Trenton, NJ 08650-4700
I have found that what is 'adequate' for one opportunity may not be adequate for another. In any event, for those who are unsure of a decision between digital products based on varying specifications, I suggest you look at the end result. Take an image and scan it in on a 24 bit scanner and a 30 bit scanner, print them out on the printer(s) you would typically use and compare the results. If the 24 bit image is sufficient for your application, then buy the thing. The street price should be about $220. If you like the 30 bit converted to 24 better, then buy it. Its street price should be around $525.
As far as the comment regarding scanning 35 mm slides, I would not recommend using a flatbed scanner to scan 35mm images that will be enlarged more than 1x the original size, otherwise the image gets soft even with a 600 dpi scanner. If you are scanning 35mm on a regular basis, you should use a film scanner.
John D. Warren Area Sales Manager Digital Products Polaroid Corporation
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
On Wed, 12 Mar 1997 WARRENJ1-at-cliffy.polaroid.com wrote:
} The 30 bits will give you additional shadow and highlight detail that } 24 bits will not provide. Now, you may ask 'my software can only } handle 24 bits so what good does 30 bits do when I will have to } convert it down?'. When you convert the file to 24 bits, the system } will choose from the best 24 bits to produce the converted file. I } would equate it to a survey or poll - the larger your sample, the more } accurate your results.
This statement seems a little bit tricky. 24 color bits gives one 256 grey shadows. If one want only to store and reproduce images - this is more than enough (128 is O'K for viewing). If one want to quantify grey levels for processing, 256 levels is also usually enough because of narrow linear range of film, grain, etc. Interpolation will give better results. If one want to waste money - I have no reasons not to this :)
Hey, y'all Just wanted to thank everyone who replied to my request for recent job postings. Lots of people went digging through their trash and came up with the information that I needed. Thanks very much, Beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
RE: David R. Stadden's inquiry (Where can I get one of those red antistatic guns, now that they are no longer handled by Fullam? Does anyone know of the English company that has this huge supply? Are there any distributors? Is there any similar product?)
I hope this is useful: Sigma Chemical Company (St. Louis, Missouri) carries the Zerostat 3 (approx $55; US catalog #Z10,881-2). Their UK contact info (from the list in the US catalog) is as follows:
In reguards to the question of tonicity, does anyone know the best percentage of sucrose to have in a primary Zamboni fixative(2% para and 15% picric acid in sorensons) to fix cell cultures in order to minimize any cell distortion.
Thanks in advance
Bob Underwood Morphology core University of Washington
In reguards to the question of tonicity, does anyone know the best percentage of sucrose to have in a primary Zamboni fixative(2% para and 15% picric acid in sorensons) to fix cell cultures in order to minimize any cell distortion.
Thanks in advance
Bob Underwood Morphology core University of Washington
} Our resident computer guru says that we should go with the cheaper } version because we are unlikely to have an output source with better } than 600 dpi for most stuff. It's true; the printers around here are } mostly 300 - 600 dpi. Is there a compelling reason for the better } model? Will I be really sorry in a year if I don't?
One reason to go with the more expensive one is that the 30bit depth will get you more contrast discrimination. Another reason is that, even with 300-600dpi printers, you might want to select a subregion of the scan and expand it to full page. In that case, the extra resolution of the better scanner will show.
At 05:04 PM 3/12/97 +1200, Richard Easingwood wrote: } Does anyone who performs vascular casting know where to obtain a product } called Mercox CL2? A fellow electron microscopist has heard that it is the } latest thing in vascular casting when mixed with methyl methacrylate but } hasn't been able to get any further info. } If anyone knows who makes it and where it can be bought we would like to hear.
There is a "Mercox Resin" in the Ladd catalog, but it doesn't say whether it's CL2.
Best regards, Steven Slap ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
Can formvar powder absorb water and cause holes in grid coatings, or is it just the solvent in which it is made that causes the problems? We have some rather old powder here and I wonder if it is still useable.
We recently purchased a film scanner to digitize images and diffraction patterns from 3.25"x4" negatives. Prior to this we had an old Microtek flatbed with a transparency adapter (300 dpi). It was ok as long as we didn't try to enlarge the image much.
I don't know how you will be using the digitized images. If all you want to do is digitize the negatives and then print the images without enlarging them, you don't need a high-resolution scanner. But I would recommend that you get a scanner with a larger bit-depth.
We often need to do some significant image analysis, so we needed a scanner which would give us very high optical resolutions and more than the usual 256 gray scale (24-bit color). We wanted at least 12 bits per color and 2400 dpi optical resolution. Our requirements may be more than you need.
I think John Warren's recommendation is good: try before you buy.
Russell E. Cook Scientific Associate Electron Microscopy Center for Materials Research Argonne National Laboratory Building 212 9700 South Cass Avenue Argonne, IL 60439 (630)252-7194 FAX: (630)252-4798
thanks to all who answered (either directly or via the listserver) my request for info on SIP refurbishment. I'll summarise these responses and make it available for anyone who contacts me directly. If there is enough interest I'll post the summary on the listserver. Cheers,
Mark Blackford TEM Group Materials Division, ANSTO PMB 1, Menai, N.S.W. Australia 2234 Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
Always get the best you can afford. And, yes, you will be sorry if you don't! One of the uses I have found for the high resolution end of the scanner is to check small details (eg is that cell junction really tight?) in an EM negative without the hassle of a large print - it's amazing just how much it is possible to see on the computer in a fraction of the time. I have the ScanMaker 3 and am very happy with it. BUT, it is no good for 35mm, only prints and large format negatives.
Diana van Driel Dept Ophthalmology Sydney University C09 AUSTRALIA 2006
} In reguards to the question of tonicity, does anyone know the best } percentage of sucrose to have in a primary Zamboni fixative(2% para and } 15% picric acid in sorensons) to fix cell cultures in order to minimize } any cell distortion.
I can't give a specific answer, but check out Maser MD et al: Relationships among pH, osmolality and concentration of fixative solutions, Stain Technology 42:175-182 (1967). You need to know what osmolality you want in the fixative solution. Around 300 milliosmols (range 200-400) seemed to be most common when I had to work it out too many years ago. In general the contribution of the fixative can be ignored. So, work out the osmolality of the buffer and add sucrose to bring the osmolality up to the total. For instance, Sorensen's is approx 105 at 0.05M, approx 210 at 0.1M. Sucrose is obvious (eg 0.2M = 200). It may be an idea to add some CaCl2 or MgCl2.
Diana van Driel Dept Ophthalmology Sydney University C09 AUSTRALIA 2006
Best practice is to dry the powder at 60 deg.C for half an hour just before mixing it into the solvent. As for the solvent you ought to fractionate it first. Redistil it and monitor the temperature of the vapour at the stillhead and discard the (watery) fraction that comes off before the correct boiling point of the solvent is reached. Stop the still when there is around an (oily) tenth remaining of the original solvent. }
We supply the Zerostat, but in the US you could get one from Sigma. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 300+ Links, MSDS ************************ http://www.proscitech.com.au }
I know this question has been asked here before, so bear with me. Where } can I get one of those red antistatic guns, now that they are no longer } handled by Fullam? Their representative told me that there is supposedly } a hefty supply of these units still in the U.K., and that Fullam doesn't } handle them anymore because they'd have to buy more than they could sell. } I know Discwasher used to distribute them. Does anyone know of the } English company that has this huge supply? Are there any distributors? } Is there any similar product? Appreciate any leads. } } Dave Stadden } DRStad-at-Juno.com
Peggy Bisher wrote: ================================================= I am looking for 3.05mm rings that I can use as a TEM grid made of ceramic material. Does anybody know where I can find such thing? ================================================== About the closest thing about which I have any awareness would be our diamond grids and rings, 3 mm diameter. They are sized to fit into any TEM that takes a 3 mm grid. More information can be found about them on our website.
Disclaimer: These are products offered by SPI Supplies and we would stand to benefit if more people were using these diamond products.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
As several people have expressed an interest in a summary of responses to my SIP query I decided to post it to the listserver. My original posting was:
} I am about to replace the sputter ion pump elements in our two JEOL TEM's. } JEOL used to be able to recondition the elements used in their 2000FX } microscopes but no longer provide this service. Can anyone suggest another } company that does this sort of work? How long can I expect a reconditioned } unit to last compared with a new one and what sort of cost would be } reasonable given that JEOL are charging $2.5k and $4.5k (Australian } dollars) for new units? Thanks in advance for any advice you can give.
Several people recommended contacting the following company:
DUNIWAY STOCKROOM Corp. 800-446-8811 also 415-969-8811 fax 415-965-0764 1600 N. Shoreline Blvd. Mountain View CA 94043 also 1305 Space Park Way,Mountain View,CA 94043 info-at-duniway.com
Vacuum Scientific Services 44 Ellesmere St, manchester M15 4JY UK Tel. 44 161 833 9108 fax. 44 161 835 1443
Some of the comments received were:
A rebuilt pump is as good as a new one and should last as long. They open up the pump, clean it out, replace all the elements, and re-weld the case together. Prices are significantly less than buying new.
Duniway has a rebuilding service and also sell their own brand of pumps. They state that rebuilt pumps have the same performance as a new pump. If the pump has to be cut open to rebuild it, it can normally can be rebuilt 2 or 3 times. They also say that their pumps can normally be rebuilt 5-10 times. Being cut open is normally the case for small pumps. On large pumps the titanium elements can often be removed through the inlet port, allowing them to be rebuilt an unlimited number of times.
I emailed Duniway Stockroom Corporation and the following is an extract from the reply by Eroc Inman:
We do rebuild not only the elements for JEOL pumps, but JEOL actually sends all of their ion pumps to us for a complete overhaul/rebuild.
It your elements are just a standard size and configuration, like that of Varian ion pumps, then the pricing is quite easy to determine. The 150 l/s pump elements are generally US$390.00/each for rebuilding. This is the price for diode or triode configuration. I would be able to give you an absolute price once we had the opportunity to take a look at them. For now, I am quite certain that your elements are not any different than what we generally see.
I have not yet contacted the other companies. If anyone can supply an email address for them I would greatly appreciate it. I don't pretend that this is a comprehensive list of companies providing SIP refurbishment, nor do I recommend or have any affiliations with any one of them. Cheers,
Mark Blackford TEM Group Materials Division, ANSTO PMB 1, Menai, N.S.W. Australia 2234 Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
For the theory you need to look up Snell's Law and Molecular refractivity. The RI relates to the velocity of light through various substances. The difference between air and water RI results in a stick appearing angled at the interface. Transparent specimen become invisible if immersed in a liquid of identical RI. This is extensively used in police scientific work were glass fragments can be identified to come from a crime scene. Very simple, put a sample onto a microscope slide. Add a drop of RI solution. If the submersed specimen becomes invisible under the microscope, than the refractive index of the specimen is that of the RI test solutions. Some experimenting to find the right solution is required. Good luck. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 300+ Links, MSDS ************************ http://www.proscitech.com.au ---------------------------------------------------------------------------- ----- } From: Damian_Neuberger_at_RLT013-at-ccmailgw.mcgawpark.baxter.com } To: microscopy-at-Sparc5.Microscopy.Com } Subject: Refractive Index } Date: Wednesday, 12 March 1997 6:49 } } I have just had a request to determine the refractive index of zinc } pyrithione, a crystalline, colorless, transparent powder. Does anyone } know what the RI is? It's not in McCrone Atlas. I will be getting a } sample Monday and with a set of Cargille RI liquids, I will determine } the RI.
############################# Notice:
This message was found in a dead-letter box and appears to be for you. If you have already gotten a copy of this message, we beg your tolerance.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
On 03/10/97 17:21:17 you wrote: } I would like tofind someone in the S.F. Bay area that has a freeze-fracture } machine that would be available for doing samples. I have a colleague in } the area who is looking for a facility where she might be able to do some } samples. } We would be very grateful for any leads. Thanks in advance. ML } } Mei Lie Wong } Department of Biochemistry } HHMI-UCSF } Ph. 415-476-4441 Fax 415-476-1902 } email wong-at-msg.ucsf.edu
Dear Mei, Please call Nancy Smith at Cal State Hayward (main number 510-885-3000). I am sure she cal help you. Regards
Igor C. Ivanov, SIMS,Auger,TEM,SEM Senior Scientist Analytical Services RJ LeeGroup, Inc. Contract Research 530 McCormick Str. (510)-567-0480 phone San Leandro, CA 94577 (510)-567-0488 FAX
############################# Notice:
This message was found in a dead-letter box and appears to be for you. If you have already gotten a copy of this message, we beg your tolerance.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thanks to all who responded to my cpd inquiry. The consensus seems to be that fresh, dry 100% ETOH is very important and that I need to keep my samples longer in the liquid CO2 to get a better exchange with the ETOH.
I'll try out all the suggestions. Hopefully, the only raisins I'll get from now on will be in my breakfast cereal. = )
Thanks again.
Paula = )
p.s. I'm going to phone some vendors of the resins & such to see if they've done studies as to which type of gloves work best. I'll keep you posted.
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
############################# Notice:
This message was found in a dead-letter box and appears to be for you. If you have already gotten a copy of this message, we beg your tolerance.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Duniway Stockroom Corp. (800-446-8811; info-at-duniway.com) advertise a service for rebuilding sputter ion pumps (p. 25 of their catalog), and state that a properly rebuilt pump should "have the same performance as a new pump". The cost of rebuilding varies from about $400 to about $3000, depending on the size and type of pump No commercial interest - just trying to be helpful.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
############################# Notice:
This message was found in a dead-letter box and appears to be for you. If you have already gotten a copy of this message, we beg your tolerance.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Dr. Nunes:
Your question, "how to stain enzymes for EM" is so broad as to be unanswerable, I think. Which enzymes?, Plant tissue or animal? Cells grown in culture? I am sure someone on the list can help if your question is more specific.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (908)-235-4583; fax -4029 e-mail: mcauliff-at-.umdnj.edu *************************************************************** ############################# Notice:
This message was found in a dead-letter box and appears to be for you. If you have already gotten a copy of this message, we beg your tolerance.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Does anyone who performs vascular casting know where to obtain a product called Mercox CL2? A fellow electron microscopist has heard that it is the latest thing in vascular casting when mixed with methyl methacrylate but hasn't been able to get any further info. If anyone knows who makes it and where it can be bought we would like to hear.
Thanks in advance
Richard Easingwood South Campus Electron Microscope Unit School of Medical Sciences University of Otago PO Box 913 Dunedin NEW ZEALAND
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Duniway Stockroom Corp. (800-446-8811; info-at-duniway.com) advertise a service for rebuilding sputter ion pumps (p. 25 of their catalog), and state that a properly rebuilt pump should "have the same performance as a new pump". The cost of rebuilding varies from about $400 to about $3000, depending on the size and type of pump No commercial interest - just trying to be helpful.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
############################# Notice:
This message was found in a dead-letter box and appears to be for you. If you have already gotten a copy of this message, we beg your tolerance.
In the theme of Diana van Driel's post check out Arborgh et al. "The osmotic effect of glutaraldehyde during fixation". J. Ultrastr. Res. 56:339-350, 1976. A very interesting and through study.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (908)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
Thanks to all for your replies concerning measuring the refractive index of a crystalline powder using a refractometer. The replies indicated that it is not easy and has very specific requirement that I can not obtain. So I will use a set of standard refractive index liquids and a polarized light microscope which is the way I had originally planned to go.
Thanks again for the comments, you folks are a veritable deep well of knowledge.
Some time back, the Snappy framegrabber board, from Play, Inc. was recommended as an economical way to obtain digitized EM images. Well, acting on this recommendation, I purchased a Snappy board. However, I have been unable to obtain an acceptable stored image file, using the Snappy board (and it's associated software) in conjunction with a CCD camera and macro lens mounted above a light box on which the EM negative is mounted. Previously, this technique has worked well, when using another grabber board (costing around five times as much!) and NIH Image software. Has anyone obtained reasonable captured images of this type using the Snappy board? I fear that this application is beyond the resolution capabilities of the Snappy board, but there is always the possibility that I'm not setting something properly in the Snappy capture software.
Over the years I've found that uranyl acetate (dry powder/crystals) seem to degrade with time and find the staining greatly attenuated. It becomes less soluble and keeps for a shorter time in solution. Does anyone know the mechanism(s) of this change and how to perhaps prevent it? I do store dark, and have had it happen in both plastic and glass containers. I make my staining solutions up in small volumes. I've ended up having more picked up by EH&S than I've ever really used. If it's inevitable, maybe we could ask our vendors to offer smaller quantities as an alternative......Grace Kennedy
Please do not reply via eMail but direct your resume to: M R Jobe Salaried Personnel THE GOODYEAR TIRE & RUBBER COMPANY 1144 E. Market Street Akron, Ohio 44316
The Goodyear Tire & Rubber Company, a world leader in tire development and manufacturing, has an excellent opportunity for a professional skilled in Microscopy in its Analytical Sciences Department, Corporate Research Division, located in Akron, Ohio. This position involves the analysis and characterization of polymers and materials used in the tire and rubber industry.
Qualified candidates must have a PhD or MS with 3 years experience in light, electron and atomic force microscopy with a strong background in image analysis. Current experience should include techniques employed in polarized light, brightfield, phase contrast, fluorescence, interference microscopy and photomicrography. Knowledge of transmission and scanning electron microscopy and EDS systems is also required. Familiarity with failure analysis and mixing studies of rubber, plastics and composites is desirable.
This position with a Fortune 100 industry leader offers excellent benefits, relocation assistance and competitive salary commensurate with education and experience. If you have the qualifications and desire to meet the rewarding challenges presented by Goodyear, direct your resume to:
M R Jobe Salaried Personnel THE GOODYEAR TIRE & RUBBER COMPANY 1144 E. Market Street Akron, Ohio 44316
An Equal Opportunity Employer, M/F/D/V Applicants must be lawfully authorized to work in the U.S.
L E Porter Phone (330) 796-1620 Head of Microscopy Fax (330) 796-3304 The Goodyear Tire & Rubber Company EMail LPORTER-at-GOODYEAR.COM Dept 415A 142 Goodyear Blvd Akron, OH 44305 USA
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Good Afternoon (Central Std Time USA) all Microscopists:
I have just had a request to determine the refractive index of zinc pyrithione, a crystalline, colorless, transparent powder. Does anyone know what the RI is? It's not in McCrone Atlas. I will be getting a sample Monday and with a set of Cargille RI liquids, I will determine the RI.
Alternatively, I have a B & L Abbe type refractometer used to determine refractive index of liquids. However, I thought I read somewhere that it could be used to determine the RI of solids. If this is in fact possible, does anyone out there know how?
My wife is having to do a poll of scientist for one of her certification classes. She has taught science in middle and high school. For the philosophers in the crowd I hope that you can have fun with this one, if you could help out with her survey it would be much appreciated. For all others, I ask your indulgence. Please forward you replys to me and I will deliver them. The survey follows.
Thank you, Chuck
SURVEY QUESTIONS
Name: Degree/Profession:
1. How would you describe the Nature of Science?
2. Describe the ideal classroom and curriculum for scientific learning.
3. What role do you believe the History of Science should play in teaching science in the classroom?
END SURVEY ------------------------------------- Name: Charles Gilbert VOC:(704)355-5261 Carolinas Medical Center FAX:(704)355-7996 Dept of Pediatric Research PO Box 32861 Charlotte, NC 28232-2861
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am acquiring an ISI SX-30E and need a copy of the manual for this instrument without paying the hundreds of dollars Topcon wants for such a manual. Any suggestions where I might be able to get a xerox of the manual or borrow a copy to xerox. Any help would be appreciated.
Jim Ekstrom Phillips Exeter Academy
############################# Notice:
This message was found in a dead-letter box and appears to be for you. If you have already gotten a copy of this message, we beg your tolerance.
Hello Fellow Listserv Members, For the past 15 or so years I have been using a wonderful film by Kodak which makes very nice black and white slides of continous tone and high contrast copy. This is a positive film which requires no reversal processing. It is called Kodak Direct Duplicating Microfilm 2468. I have been buying it from a supplier in Florida who has apparently gone out of business. Now here is the catch. Kodak, according to my long time photography needs supplier, is not willing to sell this film in anything less than a case. There are 50 100ft rolls in a case at about $40.00 per 100ft roll. My question is....are there any of you out there who might be familiar with this film and know of suppliers other than Brandon's in Jacksonville, FLA.? Brandon's is the supplier from whom I have ordered this film in the past. Or another question which has just come to mind...are there any EM Suppliers out there who might be interested in this film as one of their catalogue items? I would appreciate hearing from any of you who might have information on where to find this film or someone who might be interested in purchasing a case for distribution. I trust this is an appropriate post. Since we all make slides from time to time, I felt that it might be. Perhaps it would be better to reply to me directly and if there are others interested, let me know and I will pass on any information I receive. Thank you very much, Sandra Zane Sandra F. Zane, EM Tech. sfzane-at-email.uncc.edu Dept. of Biology, UNCC Ph.(704)547-4051 9201 University City Blvd. Fax (704)547-3128 Charlotte, NC 28223
We just received a Polaroid Sprintscan 45 for digitizing negatives from our microscopes. Preliminary trials look good. Unfortunately, no negative holder is available for TEM negatives (3 1/4 x 4 inch). This is really unfortunate since we have hundreds of such negs to scan and using the 4x5 holder isn't very user friendly - although it can be done. Has anyone had experience with this system? Any workarounds?
I suggested to Polaroid that if they would provide me with a couple of their standard 2 1/4 x 2 3/4 inch adapters, that I could have our shop mill out the proper size frame. I was shocked to find that not only were no plans in place to make the TEM holders but that Polaroid would not provide even the stock holders ( or even the metal) for modification. Caveat emptor.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 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 ####################################################################
About 6 months ago I asked the group a similar question about the Snappy and got a number of good responses concerning quality of cable, sources of noise, etc.
I have sent a rather long summary of all these replies to the originator of this post, and just wanted to make everyone else aware it is available. Just E-mail me and I'll forward it on to you individually (didn't want to bother the whole listserve with such a long message).
-Karen
On March 14, 1997, demczyk-at-erxindy.rl.plh.af.mil wrote:
} Some time back, the Snappy framegrabber board, from Play, Inc. } was recommended as an economical way to obtain digitized EM images. } Well, acting on this recommendation, I purchased a Snappy board. However, } I have been unable to obtain an acceptable stored image file, using } the Snappy board (and it's associated software) in conjunction } with a CCD camera and macro lens mounted above a light box on which the } EM negative is mounted. Previously, this technique has worked well, } when using another grabber board (costing around five times as much!) } and NIH Image software. Has anyone obtained reasonable captured images of } this type using the Snappy board? I fear that this application is beyond } the resolution capabilities of the Snappy board, but there is always the } possibility that I'm not setting something properly in the Snappy capture } software. } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Life Sciences Sector Lab Reply: kszaruba-at-mmm.com 3M Company 3M Center 270-1S-01 Phone: 612-737-2971 St. Paul, MN 55144-1000 Fax: 736-1519
These opinions are my own and may not represent those of 3M.
I am sorry that you were given the information that you were given. I recommended earlier this year that we offer a carrier for TEM negatives due to the lack of other currently available options. The last word I heard was that we are working on a carrier that will be adjustable for various film sizes. They also are working on creating a carrier dedicated to the TEM format you mentioned. As a temporary solution, cutting a mask that would fit in the 4x5 holder with the proper opening should work.
I am going to forward your message to the Worldwide Product & Technical Managers for Polaroid Scanners.
I'll keep you posted.
John D. Warren Area Sales Manager Digital Products Polaroid Corporation
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We just received a Polaroid Sprintscan 45 for digitizing negatives from our microscopes. Preliminary trials look good. Unfortunately, no negative holder is available for TEM negatives (3 1/4 x 4 inch). This is really unfortunate since we have hundreds of such negs to scan and using the 4x5 holder isn't very user friendly - although it can be done. Has anyone had experience with this system? Any workarounds?
I suggested to Polaroid that if they would provide me with a couple of their standard 2 1/4 x 2 3/4 inch adapters, that I could have our shop mill out the proper size frame. I was shocked to find that not only were no plans in place to make the TEM holders but that Polaroid would not provide even the stock holders ( or even the metal) for modification. Caveat emptor.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 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 ####################################################################
Have you tried Polaroid's 35mm Polagraph Instant Slide Film? It is a high contrast b&w film that you can process & mount at your desk. The processing does require our processor - either power or manual. The film comes with the development chemistry. You just have to buy the reusable slide mounts separately. It is a little denser than wet processed film, but it may not be as apparent with a high contrast image.
John Warren Area Sales Manager Digital Products Polaroid Corporation
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello Fellow Listserv Members, For the past 15 or so years I have been using a wonderful film by Kodak which makes very nice black and white slides of continous tone and high contrast copy. This is a positive film which requires no reversal processing. It is called Kodak Direct Duplicating Microfilm 2468. I have been buying it from a supplier in Florida who has apparently gone out of business. Now here is the catch. Kodak, according to my long time photography needs supplier, is not willing to sell this film in anything less than a case. There are 50 100ft rolls in a case at about $40.00 per 100ft roll. My question is....are there any of you out there who might be familiar with this film and know of suppliers other than Brandon's in Jacksonville, FLA.? Brandon's is the supplier from whom I have ordered this film in the past. Or another question which has just come to mind...are there any EM Suppliers out there who might be interested in this film as one of their catalogue items? I would appreciate hearing from any of you who might have information on where to find this film or someone who might be interested in purchasing a case for distribution. I trust this is an appropriate post. Since we all make slides from time to time, I felt that it might be. Perhaps it would be better to reply to me directly and if there are others interested, let me know and I will pass on any information I receive. Thank you very much, Sandra Zane Sandra F. Zane, EM Tech. sfzane-at-email.uncc.edu Dept. of Biology, UNCC Ph.(704)547-4051 9201 University City Blvd. Fax (704)547-3128 Charlotte, NC 28223
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Does anyone who performs vascular casting know where to obtain a product called Mercox CL2? A fellow electron microscopist has heard that it is the latest thing in vascular casting when mixed with methyl methacrylate but hasn't been able to get any further info. If anyone knows who makes it and where it can be bought we would like to hear.
Thanks in advance
Richard Easingwood South Campus Electron Microscope Unit School of Medical Sciences University of Otago PO Box 913 Dunedin NEW ZEALAND
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thanks to all who responded to my cpd inquiry. The consensus seems to be that fresh, dry 100% ETOH is very important and that I need to keep my samples longer in the liquid CO2 to get a better exchange with the ETOH.
I'll try out all the suggestions. Hopefully, the only raisins I'll get from now on will be in my breakfast cereal. = )
Thanks again.
Paula = )
p.s. I'm going to phone some vendors of the resins & such to see if they've done studies as to which type of gloves work best. I'll keep you posted.
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
############################# Notice:
This message was found in a dead-letter box and appears to be for you. If you have already gotten a copy of this message, we beg your tolerance.
Does anyone have any experience with this microscope. Is this considered a fine microscope for the microscopy hobby? Is this scope lacking any of the essentials or necessities for the amateur microscopist? I was surfing the net under microscopes and this one really caught my attention. Any comments would be greatly appreciated.
A few days ago someone asked about having sputter ion pumps rebuilt. It occurs to me to comment that sometimes it is possible to squeeze a few months of extra use out of a pump before rebuilding becomes absolutely necessary. Frequently, pumps become unstable because a low impedence path develops between the anode and cathode structures, usually because a flake of Ti breaks off the anode and lodges between it and the cathode, or because of the growth of a Ti whisker between the two electrodes. Sometimes such a condition can be relieved temporairly by turning the power to the pump on for a few seconds (ONLY a FEW) when the pressure in the pump is just above 1 Pa (0.01 Torr), repeating the process three or four times, if necessary. (See Vac. Methods in EM, p 295) If this treatment is successful, the pump's performance will stabalize, and the pump will perform satisfactorily again, but only for a limited time - ultimately, of course, it will need to be reconditioned.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
Hi, Help! I just started to use the NIH-image to analyze the distribution of gold particles on the EM negatives. The software was downloaded from the web site and it is NIH 1.61. My problem at this point is that I can not open my scanned image to the proper size I wanted. I tried to acquire, open or import the image, the result is the same -- a very small picture show up on the screen. When this image was magnified, the pixels came out which make particle counting impossible. Apparently, it is not the problem with the scanner since the image could be processed nicely with Adobe. But if I open the Adobe processed image through NIH, the same thing happens as described above. I also tried to scan the negatives using higher ppi, approximately 1000 ppi to incease the resolution. That did not help. Any suggestions and help will be greatly appreciated.
When I label LR White thin sections of decalcified bone with immunogold I get a fine contamination - dirt - that is not present on my conventional EM/epon sections. The contaminant is a fine floccular stuff made up of filament- like strands, each less than 10 nm in diameter, that clump togethr into irregular shapes of widely ranging size. The contaminant is not seen on the phosphorescent viewing screen but definitely shows up in the negative and print. All diluents and wash solutions are filtered with 0.2 micron Gelman Acrodisc syringe filters. Antibodies are diluted either in Tris- saline or BioMeda Primary Antibody Diluent; blockers are serum, fish gelatin, BSA. Wash solutons ar tris-saline. All solutions have Triton X. Final staining is with methanolic or aqueous uranyl acetate. Deleting uranyl acetate staining still leaves contamination. Deleting primary antibody still leaves contamination. LR White alone without processing for immuno and stained with uranyl acetate is clean. So, something in the gold, or in the secondary antibody may be doing it, but I haven't checked this out yet. Any ideas on what this dirt could be and how to get rid of it would be appreciated. Sorry for the long-windedness...and Thanks.
Pat Masarachia Bone Biology Merck Research Labs West Point, PA tel:215-652-7999 e-mail: pat_masarachia-at-merck.com
The Central Microscopy Research Facility at the University of Iowa will host an immunocytochemistry workshop given by Jan Leunissen and Peter Van der Blass. It is to be held May 9-10, 1997 in Iowa City. For more information, visit our website at: http://www.uiowa.edu/~cemrf/cemrf/jan_workshop.html or contact Kenneth Moore (319)-335-8143, kenneth-moore-at-uiowa.edu
Randy Nessler rnessler-at-emiris.iaf.uiowa.edu Views expressed are my own.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
NIH Image is extensively used in the Integrated Microscopy Resource (IMR). We never have this kind of problem. If you want, I can FTP you some of my images. In this way you can easily find where the problem is. Hope that will help you.
Ya Chen
Ya Chen
========================================================================= \ / Integrated Microscopy Resource (IMR)-- \ / __ an NIH Biomedical Research Resource TEL : 608-263-8481 \/ / / University of Wisconsin-Madison FAX : 608-265-4076 / / / 1675 Observatory Drive #159 Email1:ychen14-at-facstaff.wisc.edu / /__/_ Madison, WI 53706 Email2:chen-at-calshp.cals.wisc.edu ========================================================================= IMR WWW Home Page: http://www.bocklabs.wisc.edu/imr.html
Does anyone have available (new or used) a double tilt holder, a double tilt heating holder, a double tilt cooling holder and a selective aperture set for the LEO/Zeiss 910 TEM? If so, please let me know (incl. price).
Sandra Zane was wondering where she could get Kodak Direct Duplicating Microfilm 2468. We use Eastman 5360 which sounds like the same thing; 35 mm, gives a positive without a complicated kit, developes in Dektol. We order it locally (I think) or you can get it from Freestyle Sales Co. 5124 Sunset Blvd. Los Angeles, CA 90027, www.freestylesalesco.com. In their catalogue it is called "Kodak B&W duplicating film 5360" and is available in 50 or 100 foot rolls or 4x5 sheets.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (908)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
POSTDOCTORAL POSITION is available immediately to develop three dimensional image analysis algorithms for quantitative analysis of cells in tumor specimens. Experience in image analysis and UNIX workstations, and a strong math background are essential. Candidate should have a recent Ph.D. in the physical sciences, or Computer Science and should be an expert in developing code in C/C++. Send curriculum vitae and the names of three referees to:
Dr. Ravi Malladi, MS 50A-2152 Lawrence Berkeley National Laboratory University of California, Berkeley, CA 94720.
Forgive my inexperiece and knowledge on this new hobby of mine but can someone briefly explain the differece between semi-plan objectives and plan objectives. Under what applications would one type be more advantageous over the other type. Thanks for any information you may have on this subject.
Hope I'm not duplicating an earlier topic but couldn't find anything in postings kept over the last few months so, here goes...
What factor (s) in an immungold labeling protocol has (have) the greatest impact on background, e.g.. antibody dilution, buffer, etc.? I'm looking at the immunogold protocol itself so I can use valuable tissue already embedded.
(I'm labeling virus-infected plant leaf tissue with monoclonal antibodies to viral proteins - samples are embedded in LR White and fixed in 3% paraformaldehyde/0.2% glutaraldehyde. We block before primary antibody incubation with a TRIS-HCl buffer containing bovine serum albumin and goat normal serum. Our secondary antibody is a commercially prepared goat anti-mouse gold conjugate. Usually we have excessive background on chloroplasts, mitochondria and cell walls; if we get rid of background by increasing dilutions of antibodies, we lose specific reactions as well).
One interesting point....we don't have this problem with polyclonal antibodies, only monoclonals.
Thanks for any tips -- this is really frustrating!
Peggy
Peggy Brannigan Electron Microscopy Floral and Nursery Plants Research Unit National Arboretum
} Date: Mon, 17 Mar 1997 16:33:06 -0800 } From: Geoff McAuliffe {mcauliff-at-UMDNJ.EDU} } To: MSA Listserver {Microscopy-at-Sparc5.Microscopy.Com} } Subject: re: positive film } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear List: } } Sandra Zane was wondering where she could get Kodak Direct } Duplicating Microfilm 2468. We use Eastman 5360 which sounds like the 0} same thing; 35 mm, gives a positive without a complicated kit, developes } in Dektol. We order it locally (I think) or you can get it from Freestyle } Sales Co. 5124 Sunset Blvd. Los Angeles, CA 90027, } www.freestylesalesco.com. In their catalogue it is called "Kodak B&W } duplicating film 5360" and is available in 50 or 100 foot rolls or 4x5 } sheets. } } Geoff } -- } *************************************************************** } Geoff McAuliffe, Ph.D. } Neuroscience and Cell Biology } Robert Wood Johnson Medical School } 675 Hoes Lane Piscataway, NJ 08854 } voice: (908)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu } *************************************************************** } We have good success with 5360 also, and it's easy to get. Here is a copy of information I sent directly to Sandra:
The Direct MP Film 5360 is cat # 24611 from Ted Pella, $22.90/100 ft roll. 800 237-3526 or tedpel-at-aql.com or Fax 916 243-3761 I put the camera on B and manually work the shutter for 2, 2.5, 3 sec, depending on the size (height of camera). It's so cheap, I usually bracket and shoot 3 shots for each to make sure I don't have to repeat shooting. Most of the time, any of the shots could be used, but I pick the ones that are most closely matched for use in a single presentation. I develop in D-19 4 min at 20 oC. There is another film, Kodak Contrast Copy Film, that works about the same, but gives a bluer tint. I only keep the 5360 now.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
I have done a lot of LR White immunostaining and have had many kinds of dirt problems. It sounds like you are doing nearly everything right but you didn't mention doing a final rinse in filtered distilled water after your gold secondary. I rinse 2-3x in my buffer after secondary then do a fairly vigorous rinse in the beaker of distilled filtered water, then dry the grids, then go to staining.
You may also check your water and photoflo source and see if it is actually on your negatives.
Bob Morphology Core U.of Washington Seattle
On Mon, 17 Mar 1997, Pat Masarachia wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } When I label LR White thin sections of decalcified bone with } immunogold I get a fine contamination - dirt - that is not present on } my conventional EM/epon sections. } The contaminant is a fine floccular stuff made up of filament- } like strands, each less than 10 nm in diameter, that clump togethr } into irregular shapes of widely ranging size. The contaminant is not } seen on the phosphorescent viewing screen but definitely shows up } in the negative and print. } All diluents and wash solutions are filtered with 0.2 micron } Gelman Acrodisc syringe filters. Antibodies are diluted either in Tris- } saline or BioMeda Primary Antibody Diluent; blockers are serum, fish } gelatin, BSA. Wash solutons ar tris-saline. All solutions have } Triton X. Final staining is with methanolic or aqueous uranyl acetate. } Deleting uranyl acetate staining still leaves contamination. } Deleting primary antibody still leaves contamination. LR White alone } without processing for immuno and stained with uranyl acetate is clean. } So, something in the gold, or in the secondary antibody may be } doing it, but I haven't checked this out yet. Any ideas on what this } dirt could be and how to get rid of it would be appreciated. } Sorry for the long-windedness...and Thanks. } } Pat Masarachia } Bone Biology } Merck Research Labs } West Point, PA } tel:215-652-7999 } e-mail: pat_masarachia-at-merck.com } } }
I would think if your minus primary control are clean then your primary antibody is non-specificly binding to something in the plant tissue. Usually the primary has the most effect on backround. Is your antisera purified?
Bob Morphology Core U of Washington Seattle
On Tue, 18 Mar 1997, Peggy Brannigan wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello immunogold experts! } } Hope I'm not duplicating an earlier topic but couldn't find anything in } postings kept over the last few months so, here goes... } } What factor (s) in an immungold labeling protocol has (have) the greatest } impact on background, e.g.. antibody dilution, buffer, etc.? I'm } looking at the immunogold protocol itself so I can use valuable tissue } already embedded. } } (I'm labeling virus-infected plant leaf tissue with monoclonal antibodies } to viral proteins - samples are embedded in LR White and fixed in 3% } paraformaldehyde/0.2% glutaraldehyde. We block before primary antibody } incubation with a TRIS-HCl buffer containing bovine serum albumin and goat } normal serum. Our secondary antibody is a commercially prepared goat } anti-mouse gold conjugate. Usually we have excessive background on } chloroplasts, mitochondria and cell walls; if we get rid of background by } increasing dilutions of antibodies, we lose specific reactions as well). } } } One interesting point....we don't have this problem with polyclonal } antibodies, only monoclonals. } } Thanks for any tips -- this is really frustrating! } } } Peggy } } Peggy Brannigan } Electron Microscopy } Floral and Nursery Plants Research Unit } National Arboretum } } Bldg. 010A R.238 } 10300 Baltimore Avenue } Beltsville, MD USA20705 } } Phone: (301) 504-6097 } Fax : (301) 504-5096 } Email: brannign-at-asrr.arsusda. gov } } } }
We also find that immuno-EM is problematic with MAbs. Are you using ascites or an IgG prep? I would use IgG if possible (the EZ Sep products from Pharmacia Biotech are handy for isolating IgG from ascites, MHO). You may want to try adding 0.05% Tween-20 to your blocking and antibody soln's. This often helps bring down the background. The downside (there's always a downside) is that the tween also tends to wash a lot of the contrast out of the sections -- at least it does for our LRGold embedded material. You may need to adjust the length of time you let the grids float on the antibody sol'n to give good label without too much loss of contrast. 1-4 hours on the primary is a good range for starters.
Hope this helps,
Greg Martin Dept. of Cell Biology and Anatomy Johns Hopkins School of Medicine
On Tue, 18 Mar 1997, Peggy Brannigan wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello immunogold experts! } } Hope I'm not duplicating an earlier topic but couldn't find anything in } postings kept over the last few months so, here goes... } } What factor (s) in an immungold labeling protocol has (have) the greatest } impact on background, e.g.. antibody dilution, buffer, etc.? I'm } looking at the immunogold protocol itself so I can use valuable tissue } already embedded. } } (I'm labeling virus-infected plant leaf tissue with monoclonal antibodies } to viral proteins - samples are embedded in LR White and fixed in 3% } paraformaldehyde/0.2% glutaraldehyde. We block before primary antibody } incubation with a TRIS-HCl buffer containing bovine serum albumin and goat } normal serum. Our secondary antibody is a commercially prepared goat } anti-mouse gold conjugate. Usually we have excessive background on } chloroplasts, mitochondria and cell walls; if we get rid of background by } increasing dilutions of antibodies, we lose specific reactions as well). } } } One interesting point....we don't have this problem with polyclonal } antibodies, only monoclonals. } } Thanks for any tips -- this is really frustrating! } } } Peggy } } Peggy Brannigan } Electron Microscopy } Floral and Nursery Plants Research Unit } National Arboretum } } Bldg. 010A R.238 } 10300 Baltimore Avenue } Beltsville, MD USA20705 } } Phone: (301) 504-6097 } Fax : (301) 504-5096 } Email: brannign-at-asrr.arsusda. gov } } } }
I am looking for a person to work on an exciting new project.
The appointment could be at the post-doctoral level or at other levels according to the background of the person appointed. In any case the post will be for three years.
The job is at the Materials Research Laboratory of the University of Illinois at Urbana.
The project is a joint enterprise involving Argonne National Lab, Oak Ridge National Lab, the Lawrence Berkeley Lab and NIST as well as the University of Illinois. The Project has the aim of developing a new kind of environment for electron microscopy and related techniques, in which the instruments can be operated remotely with the same effectiveness as they can be operated in the instrument room. More details of the project can be found at http://tpm.amc.anl.gov/MMC MMC is the abbreviation of the project name.
I am looking for someone who has familiarity with electron microscopy (preferably TEM) or a closely related technique - and who has well developed interests and experience in computing, particularly the interfacing of instruments for computer control and/or the networking of images.
Will any one interested please contact me right away. We would like the job to be started as soon as possible. ** Alwyn Eades Center for Microanalysis of Materials University of Illinois at Urbana-Champaign Phone 217 333 8396 Fax 217 244 2278 eades-at-uimrl7.mrl.uiuc.edu (NB those are letter l not ones) **
You are invited to attend the 1997 Annual Spring meeting of the Arizona Imaging and Microanalysis Society. The meeting will be Thursday March 27th and Friday March 28, 1997. Meetings will be held in the Senior Ballroom of the University of Arizona's Student Union Building. Admission is free to interested participants.
Please pass this information on to interested colleagues.
========================================================================= DIGITAL IMAGING =========================================================================
Digital imaging has found its way into many scientific fields (biology, engineering, astronomy, radiology, etc.). As a wider range of researchers begin to use these techniques, they are often unsure of where to begin and are unaware of the pitfalls involved in the field of digital imaging. This workshop will introduce you to some important concepts and issues relevant to digital imaging.
Thursday, March 27, 1997
9:00 Welcome and overview of the meeting Doug Cromey, President, AIMS
9:15 Image acquisition: General approaches, common problems and issues relating to CCD cameras. Scott Sternberg, Photometrics
10:15 Coffee break
10:30 Data output devices John Spenseri, Micrographix West
11:15 Data storage, movement and archiving Doug Cromey, President, AIMS
6:00 Buffet dinner in the Student Union's Sr. Ballroom (Mexican-style entrees, Preregister with Patty Jansma, cost $11.00)
Friday, March 28, 1997
9:00 Ethics and image analysis: Panel discussion. (Questions from the audience are welcome.)
Panel members: John Gilkey, UA, Pharmacology Marvin Landis, UA, CCIT- User Support David Ring, Dir. Photography, Biomedical Communications, UA Mary Rykowski, Assist. Prof., Cell Biology and Anatomy, UA Scott Sternberg, Photometrics
10:30 Coffee Break
10:45 Student platform presentations
11:45 Lunch. AIMS Business Meeting New officers installation.
1:00 Student platform presentations
3:00 Presentation of student awards and closing remarks
A histochemistry query: In the course of our work, we examine many different vertebrate tissues (brain, skin, liver, kidney, etc) embedded in plastic resin (Eponate or LR White). We currently do a lot of LM viewing of 2 micron sections, using toluidine blue as a counterstain. Some tissues have been fixed with aldehydes and osmium, for subsequent EM thin section studies. For immunocytochemistry, other tissues are only lightly fixed with aldehydes.
Our question: Can you suggest other useful counterstains, besides toluidine blue, that are compatible with plastic sections. If you have a favorite recipe, please send to us off-line, so that we don't clog the listserver. We'll compile a list of the answers we receive, for those who are interested.
Thanks in advance
David H. Hall hall-at-aecom.yu.edu Dept of Neuroscience Albert Einstein Col Medicine, Bronx, NY 10461 (718) 430-2195 (718) 430-8821 FAX
A formal proposal to create a new group tentatively called sci.bio.immunocytochem has just been posted to news.announce.newgroups. This is a reposting of that proposal.
REQUEST FOR DISCUSSION (RFD) unmoderated group sci.bio.immunocytochem
This is the 2nd Request For Discussion (RFD) for the creation of a world-wide unmoderated Usenet newsgroup sci.bio.immunocytochem, currently being discussed in news.groups
Suggestions for improvements to this proposal are welcome. Discussion about it should take place in news.groups. A vote is expected to be held in about three to four weeks.
This is not a Call for Votes (CFV); you cannot vote at this time. Procedural details are below.
CHANGES from previous RFD:
2nd RFD posted because more than 60 days have elapsed since 1st RFD. There are (minor changes in Distribution and Newsgroup line.
Newsgroup line: sci.bio.immunocytochem Immuno-labelling of biological material.
RATIONALE: sci.bio.immunocytochem
Immunohistochemists and immunocytochemists already enjoy the benefits of online communication, utilizing e-mail, accessing web sites, and subscribing to specialised mailing lists. Usenet newsgroups are also popular, but this is less obvious because articles with immunocytochemical/immunohistochemical content get posted to many different newsgroups. Most articles are posted to a favourite five or six newsgroups including bionet.cellbiol, sci.med.immunology and sci.techniques.microscopy, but often articles get posted to any one of fourteen or fifteen newsgroups in the sci. and bionet. heirarchies. Some of these are listed in the distribution list at the end of this proposal.
In my view, no existing newsgroup fulfils the criteria necessary to attract all the various immunocytochemistry postings. I do not wish to draw users away from other newsgroups, only to encourage scientists to share their knowledge and expertise on immunocytochemistry in the most effective manner. In response to my proposal to create a newsgroup dedicated to the discussion of immunocytochemistry and immunohistochemistry, I have received e-mail and faxes from researchers all over the world offering their support and encouragment.
Immunocytochemistry and immunohistochemistry are not subdivisions of immunology, molecular biology or chemistry. Microscopy, although essential, is only a small part of the story. Immunocytochemistry and immunohistochemistry are multi- disciplinary, therefore discussions are destined to stay distributed amongst the different newsgroups until they are all brought together under one umbrella. This would then act as a focus point for all the immunocytochemists who are already Internet users, and encourage new subscribers to Usenet.
CHARTER: sci.bio.immunocytochem
This is a newsgroup for the exchange of information relating to immunocytochemistry and immunohistochemistry. This unique research tool is used to locate and identify specific molecules in biological material, at the microscopical level.
Articles posted to this group must be relevant to one or more aspects of the above. The kind of subjects that may be discussed include techniques, theory, presentation of results, requests for collaboration, history, equipment, publication references, notice of events, tips and trouble-shooting, jobs offered andwanted, jokes, stories and new ideas, so long as the posting bears a direct relevance to the central theme. There will be a list of Frequently Asked Questions (FAQs) to help newcomers.
A relevant posting could just be a simple question or answer, for example "Has anyone got any experience with this reagent ?"or "Which course could I attend to learn more about immunogold labelling?". There will be articles reminding people to read the list of FAQs prior to posting their own article. Usenet readers may get involved in complex discussions about, for example, multiple labelling, proper use of control experiments, microwave antigen retrieval or quantitative measurements. Remember that articles posted to a newsgroup are intended for a wide readership, so if you have information which concerns only one or two people then please don't use this newsgroup, use e-mail.
Commercial advertisements for services, equipment or reagents violate the charter unless one or more of the following apply: (a)The advertisement is part of a comprehensive article designed specifically to address issues raised in earlier articles posted to the group (b)A general reference to the type of product does not suffice for technical reasons and it is necessary to specify the exact commercial product (c) The information is offered primarily for the benefit of the readers (d)The advertisement is for second-hand equipment specific to immunocytochemistry (e) Requests or offers for free products are acceptable if they are not part of a sales promotion.
END CHARTER.
PROCEDURE:
This is a request for discussion, not a call for votes. In this phase of the process, any potential problems with the proposed newsgroups should be raised and resolved. The discussion period will continue for a minimum of 21 days (starting from when the 2nd RFD for this proposal is posted to news.announce.newgroups), after which a Call For Votes (CFV) may be posted by a neutral vote taker if the discussion warrants it. Please do not attempt to vote until this happens.
All discussion of this proposal should be posted to news.groups. This RFD attempts to comply fully with the Usenet newsgroup creation guidelines outlined in "How to Create a New Usenet Newsgroup" and "How to Format and Submit a New Group Proposal". Please refer to these documents (available in news.announce.newgroups) if you have any questions about the process.
DISTRIBUTION:
This RFD has been posted to the following newsgroups: news.announce.newgroups,news.groups,bionet.cellbiol, bionet.diagnostics,bionet.immunology,bionet.microbiology, bionet.molbio.methds-reagnts,sci.bio.misc,sci.med.immunology, sci.techniques.microscopy
This RFD will be reposted to the following newsgroups after its posting in news.announce.newgroups: bionet.molbio.proteins,bionet.neuroscience,bionet.plants, sci.bio.microbiology,sci.med,sci.med.laboratory,sci.misc, sci.nanotech
This RFD will also be reposted to the following mailing lists after its posting in news.announce.newgroups:
Histonet mailing list: {histonet-at-pathology.swmed.edu} Information pertaining to the technical aspects of histology and histopathology such as tissue fixatives and processing, routine histology, special stains, immunohistochemistry, in-situ hybridization etc. To subscribe type "subscribe digest" into the subject box and leave the text box empty, or to subscribe to the full service just type "subscribe". For more info access web site http://www.mwrn.com/subject/histonet.htms
Microscopy Society of America listserver: Questions/comments/answers in the various fields of Microscopy Currently over 3000 subscribers. To subscribe send the message "subscribe" to {Listserver-at-MSA.Microscopy.Com} then send messages in plain text to {Microscopy-at-MSA.Microscopy.Com} For more info access web site http://www.amc.anl.gov/ Docs/anl/Nestor/Software/telecommList.html
Stanford University list server To subscribe, send a message to {majordomo-at-pathology.stanford.edu} with "subscribe ipox-l" in the body of your message. This list helps pathologists and other laboratory professionals to exchange information about immunoperoxidase methods.
This RFD will also be reposted to the following web-sites after its posting in news.announce.newgroups:
Royal Microscope Society http://www.rms.org.uk Web Master Dr R. A. D. Mackenzie {r.a.mackenzie-at-open.ac.uk}
Center for Cell Imaging Department of Cell Biology Yale University School of Medicine Introduction to Immunocytochemistry http://info.med.yale.edu/cellimg/CCIimmuno.html Web Master Paul Webster { paul_webster-at-yale.edu}
Proponent: Amanda Wilson {awilson-at-aw.u-net.com} Proponent: Paul Monaghan { monaghan-at-icr.ac.uk} Mentor: Jonathan Grobe {grobe-at-netins.net}
Amanda Wilson Deputy Manager, E.M. Unit St George's Hospital Medical School, S.W.London, UK Tel: 0181 725 5220 (work) e-mail {awilson-at-aw.u-net.com}
I am in need of an X-unit (EHT regulator) for our Philips Model 300. Does anyone happen to know where I might find a working unit? Please respond directly to me. Thanks in advance!
Scott Schwinge University of Washington Friday Harbor Labs 360-378-2165 schwinge-at-fhl.washington.edu
In our experience the following are the most important issues for good antibody staining-
quality of primary ab } fixative } buffer } dilution } blocking agents.
Most of our background problems can be corrected by getting a better antibody. Some out there work great on gels but once in among all the cellular proteins they start binding to everything.
The proper fixation (which of course varies for every different antibody antigen mix we try) is the next most critical.
We have also found differing the buffers can increase the signal to noise ratio. Although we normally use Gey's salts (it seems to works well with a whole range of antigens) sometimes cacodylate is superior.
The bottom line, however, is that with any new antigen or antibody it takes us a lot of trial and error to find the correct match.
Although we have not yet discovered a magic potion, I hope this limited answer is helpful-
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-bgsm.edu * **************************************************************
On Tue, 18 Mar 1997, Peggy Brannigan wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello immunogold experts! } } Hope I'm not duplicating an earlier topic but couldn't find anything in } postings kept over the last few months so, here goes... } } What factor (s) in an immungold labeling protocol has (have) the greatest } impact on background, e.g.. antibody dilution, buffer, etc.? I'm } looking at the immunogold protocol itself so I can use valuable tissue } already embedded. } } (I'm labeling virus-infected plant leaf tissue with monoclonal antibodies } to viral proteins - samples are embedded in LR White and fixed in 3% } paraformaldehyde/0.2% glutaraldehyde. We block before primary antibody } incubation with a TRIS-HCl buffer containing bovine serum albumin and goat } normal serum. Our secondary antibody is a commercially prepared goat } anti-mouse gold conjugate. Usually we have excessive background on } chloroplasts, mitochondria and cell walls; if we get rid of background by } increasing dilutions of antibodies, we lose specific reactions as well). } } } One interesting point....we don't have this problem with polyclonal } antibodies, only monoclonals. } } Thanks for any tips -- this is really frustrating! } } } Peggy } } Peggy Brannigan } Electron Microscopy } Floral and Nursery Plants Research Unit } National Arboretum } } Bldg. 010A R.238 } 10300 Baltimore Avenue } Beltsville, MD USA20705 } } Phone: (301) 504-6097 } Fax : (301) 504-5096 } Email: brannign-at-asrr.arsusda. gov } } } }
Im looking for a functioning TN5500 with a Dec PC preferable an 1123 or better without the Detector for a project I'm working on. Has any of you recently upgraded thier probes and is looking to sell or donate it. I will gladly pay all associated expenses. Please contact me directly at the following: Regards,
Joe Barney Micro-Analytical Service Center Inc. Phone: 717-299-0599 Fax: 717-299-2022 E-mail: Jbarjr-at-aol.com
One thing which I've found to be helpful for getting rid of non-specific background labeling is 2-3 washes with a high-salt buffer immediately following the primary Ab. I've done quite a bit of work in the past with biotinylated primaries followed by streptavidin-gold, and have seen cases of high background on occasion; it somehow seems to be primary Ab-dependent...very bizarre.
Anyway, I'm wondering if something similar might be happening in your system. If you want to give it a try, I would suggest the following:
1. Make a buffer (TRIS HCl should be fine) containing 5 normal saline. Regular normal saline is 0.9% by weight, or about 150 mM, so add enough NaCl to make it 4.5% by weight (about 750 mM). I know this sounds like a whopping dose of salt, and I can't give a complete rationale for why it works, but it *does* decrease the background!
2. Do 2-3 washes w/ the high salt following the primary, then a couple of washes w/ the normal TRIS buffer to get the ionic concentration back where it should be for the gold conjugate.
3. One other thing: Do all of your diluents contain normal goat serum (even the diluent for the gold)? If not, you might want to try this as well.
Peggy Brannigan writes: } } (I'm labeling virus-infected plant leaf tissue with monoclonal antibodies } We block before primary antibody } incubation with a TRIS-HCl buffer containing bovine serum albumin and goat } normal serum.
Peggy, we had similar problems using some home grown antisera. Our solution was to block in 2%BSA, 5% Normal X serum and 1% Skim milk powder in TBS for 1 hr. Didnt completely solve the cell wall binding but was a *huge* improvment, didnt affect the specific binding at all.
Worth a try at least --
Daryl Webb (dwebb-at-waite.adelaide.edu.au) Dept. of Plant Science, Waite Institute University of Adelaide, Glen Osmond S.A. 5064 Australia. Voice:61_8 8303 7426 Fax:61_8 8303 7102
Many thanks to everybody who replied to my query about flatbed scanners. You are all a font of all kinds of information! To summarize:
1) (Obviously) get the highest res scanner with the most pixel depth you can afford - it's hard to overdo.
2) Printers that are 300-600 dpi with dither and print grey levels at 100-200 dpi; for continuous tome they need 3x3, 4x4, or 9x9 points (depending on the type of printer) for each pixel. So the higher the resolution of the image scanned in the better the print (also obvious).
3) Opinions vary - some say that 24 bit color depth gives 256 grey levels which is "more than enough", while others say that 30 or 36 bit depth gives more contrast enhancement, even when your software only handles 24 bit color. The best bet is to try out 24 and 30 bit scanners and see which does the job for you.
4) The big problem lies in enlarging images. A 4x5 SEM negative scanned in at 300 dpi can really only become a 4x5 print. A TEM negative scanned in at 600 dpi can be enlarged {3x, whereas a TEM negative can be enlarged 10x photographically. This brings up the discussion at MSA '96 - learn to take your micrographs at the magnification at which you will want to use and reproduce them rather than enlarging and therefore gaining only "empty magnification".
5) Remember to check for the true resolution, not the interpolated res. of a scanner.
6) Be careful of getting ghosts from glossy Polaroids and Newton rings from TEM negatives on some scanners.
7) None of the flatbed scanners seem good enough for 35mm negatives - buy a film scanner instead.
8) Remember that higher res images mean larger files. WIll you be able to store them? You will soon be needing ZIPs, JAZs, and CD-Rs. (Our SEM students are alloted 10 MB storage space on their department accounts. The full-size, full-res, colorized Mexican Ant on my web site is over 8 MB by itself!)
9) See "Tips & Tricks" at http://www.biotech.ufl.edu/~emcl/ for a previous discussion on this topic.
As you can see from my "cheap frame grabber" and "cheap scanner" posts, we are trying to operate within an embarrassingly small budget this year. ALthough I am still fairly happy to prepare everything photographically for myself, when I talk about image presentation and look at the bright shining faces of my current SEM students, I know that I am not going to be teaching them darkroom techniques. They start with Polaroid 55 pos/neg film. One needs to make slides right away; another wants to know how to make camera ready illustrations for a journal by next week. Most of these students ultimately have access to some decent hardware and software, and I need to point them in the right direction. I need about $10K to bring my lab up to speed. Too bad I can't invite you all to a huli-huli chicken sale!
What brought this up? Microtek 300 dpi scanners are down to $195, which I could buy out of my own pocket. It was tempting.
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
What we need now is some advice for coating ceramic samples When to use Carbon, Gold, thickness etc.
Is there any FAQ about this available?
Any help highy appreciated.
______________________________________________________________ Andreas Loewe Tel: +49-228-734-180 University of Bonn Fax: +49-228-734-205 Insitute for Inorganic Chemistry email: loewe-at-uni-bonn.de Inorganic Material Research Roemerstr. 164 53117 Bonn Germany http://www.elmi.uni-bonn.de/ ______________________________________________________________
your problem might be quite fundamental. It's several years since I've been seriously involved with immunogold labelling but I recall that there is a basic problem with monoclonal antibodies.
If the antigenic site, which the monoclonal antibody is specific to, is particularly sensitive to fixation then it will not label, hence the lack of specific labelling. You mentioned that a polyclonal works and this could simply be because it can 'find' a less sensitive part of the antigen. Have you had a chance to try cryo or less fixation?
I apologise if my comments are out of date or irrelevant.
Malcolm Haswell University of Sunderland UK ----------
Hello immunogold experts!
Hope I'm not duplicating an earlier topic but couldn't find anything in postings kept over the last few months so, here goes...
What factor (s) in an immungold labeling protocol has (have) the greatest impact on background, e.g.. antibody dilution, buffer, etc.? I'm looking at the immunogold protocol itself so I can use valuable tissue already embedded.
(I'm labeling virus-infected plant leaf tissue with monoclonal antibodies to viral proteins - samples are embedded in LR White and fixed in 3% paraformaldehyde/0.2% glutaraldehyde. We block before primary antibody incubation with a TRIS-HCl buffer containing bovine serum albumin and goat normal serum. Our secondary antibody is a commercially prepared goat anti-mouse gold conjugate. Usually we have excessive background on chloroplasts, mitochondria and cell walls; if we get rid of background by increasing dilutions of antibodies, we lose specific reactions as well).
One interesting point....we don't have this problem with polyclonal antibodies, only monoclonals.
Thanks for any tips -- this is really frustrating!
Peggy
Peggy Brannigan Electron Microscopy Floral and Nursery Plants Research Unit National Arboretum
For x-ray analysis work use carbon (evaporate). It will (typically) attenuate less than Au and will not generate interfering x-ray lines. Because of the low Z coating and usual low z of ceramics, imaging of carbon coated ceramics should be done with as low a beam voltages as practical.
For the best imaging, use Au (sputter). It has better stopping power (= better resolution) and will provide a higher SE yield than carbon. Lower beam voltages (ie: 5-10 kv) still helpful.
Thickness: Always use the thinest coating which will prevent charging. This can sometimes be a problem on low density, friable ceramic fractures or fiberous material as a continuously conductive films can be difficult to achieve. One way to mimimize this is to paint as much of the specimen as possible (those areas not viewed) with carbon paint. Wicking can be a problem - viscosity and volitility of the paint will make quite a difference.
Alternatives (which I do not have available with my system)....
Very low kV imaging....in the area of approx. 500 to 1500 beam volts, some materials will be at an equilibrium state where energy in = energy out and thus dont't charge.
E-SEM - High chamber pressure discharges specimen and coating is not
needed. It has been my observation that this will work well for some
applications, but is surely no "cure-all". High mag, high resolution, low kV exams usually suffer.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear folks,
we recently got a new SEM.
What we need now is some advice for coating ceramic samples When to use Carbon, Gold, thickness etc.
Is there any FAQ about this available?
Any help highy appreciated.
______________________________________________________________ Andreas Loewe Tel: +49-228-734-180 University of Bonn Fax: +49-228-734-205 Insitute for Inorganic Chemistry email: loewe-at-uni-bonn.de Inorganic Material Research Roemerstr. 164 53117 Bonn Germany http://www.elmi.uni-bonn.de/ ______________________________________________________________
What we need now is some advice for coating ceramic samples When to use Carbon, Gold, thickness etc.
Andreas, it will depend ,to some extent, on what you are after. If you intend to do EDS for example, then you might want to deposit C since Au could interfere with your results. Also keep in mind that Au will deposit as relatively large islands and this will limit your resolution. You will be better off using Au-Pd for example as opposed to just Au. How thick a film you need will also depend , among other things, on the voltage that you are using in the SEM and on how flat your sample is (i.e. are you looking at a fracture surface or a polished surface ?). In most cases, at 10 or 20KeV , 10-30 nm of Au-Pd should be enough provided the sample is grounded properly (i.e. use a conductive paint, such as gold or carbon, around your sample). If you operate at lower voltages , a thinner coating might be O.K. You might want to read some standard texts on SEM and experiment a bit with your samples.
Jordi Marti ______________________________________________________________ Andreas Loewe Tel: +49-228-734-180 University of Bonn Fax: +49-228-734-205 Insitute for Inorganic Chemistry email: loewe-at-uni-bonn.de Inorganic Material Research Roemerstr. 164 53117 Bonn Germany http://www.elmi.uni-bonn.de/ ______________________________________________________________
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear folks, } } we recently got a new SEM. } } What we need now is some advice for coating ceramic samples } When to use Carbon, Gold, thickness etc. } } Is there any FAQ about this available? } } Any help highy appreciated. } } ______________________________________________________________ } Andreas Loewe Tel: +49-228-734-180 } University of Bonn Fax: +49-228-734-205 } Insitute for Inorganic Chemistry email: loewe-at-uni-bonn.de } Inorganic Material Research } Roemerstr. 164 } 53117 Bonn } Germany http://www.elmi.uni-bonn.de/ } ______________________________________________________________ } } Andreas, First make sure that the sample is well grounded. Then it will be trial and error as to how thick to coat the sample. Gold usually gives the best results.--Good Luck
} Gregory Rudomen Greg-at-UMIC.SUNYSB.EDU University Microscopy Imaging Center S.U.N.Y. Stony Brook
The undesired signals are present only when you include your monclonal antibody, if I understand correctly. I don't think that it is just the fact that you have a monoclonal, after all, antiserum is more or less merely a collection of monoclonals.
If this undesired signal is caused by an unspecific reaction at the primary antibody level, as often found e.g. with IgM type antibodies (the bigger the stickier), you would expect to be able to get rid of it in the firat place by improving the block step. This takes care of background caused by hydrophobic interactions. Secondly, the incubation buffer should have additives (like BSA, cold water fish gelatin and there is a lot more!) which compete with primary antibodies for unspecific binding. In that case the protein additive should prevent or at least reduce background. Sometimes Tween does help, but since it is a detergent I would only use it on embedded material, not on cryosections. If you have to use detergent the best results are obtained at concentrations slightly higher than the critical micelle concentration.
On the other hand, from your description I get the impression that with increasing the antibody dilution both your specific and undesired signal decrease, which may indicate that "both types" of reaction have similar affinities. The question now is: is this undesired signal background or a more or less specific signal? Your specific reaction should at least have a lower Kd than the background reactions. If that is not the case, only a different antibody with higher affinity will help.
BTW: The topic of specificity and background will be extensively dealt with at the immunocytochemistry workshop in Iowa City, announced earlier on this Listserver.
Good luck!
============================= Jan Leunissen, Ph.D. AURION ImmunoGold Reagents & Accessories Costerweg 5, 6702 AA Wageningen The Netherlands
In 1992 Lindley (Microsc. Res. & Techn. 21:355) published a technique for using a primer (Z-6040, a silane) to allow for good adherence of biological materials with outer surfaces that do not normally stick well to embedding plastics. The plastic used was LRWhite. I am interested in using this primer with Epon 812 substitutes. Has anyone used this primer with the epoxies and is there a protocol.
In a similar vein, does anyone have a good protocol for embedding cell cultures on transwell membranes with small pores, such that when sectioning the cells + membrane the membrane does not rip away from the cells tearing off the basal layer of the bottom cells. Different membrane polymer types differ have different effects, as well as different pore sizes. If a very thick collagen layer is laid on top of the membrane this helps, but for experimental work this is often undesirable.
I just had to do a job for an investigator that wanted their prints made into slides. I have usually let them them take care of that and found out that our Biomed's graphic people use color slide film only which heard gives a blue cast to the EM prints. I decided to try Kodak's RPC # 175-3151 film on our copy stand (the people I got it from use it for taking slides of X-rays). It turned out Great! The draw back.... 10 and 15 second exposure brackets, at f4 f-stop, with four 500 watt photofloods. I developed it in our Xomat x-ray processor but was told you can use D-19 also. I'm going to reshoot the prints with Fugichrome 100 and that Direct MP 5360 someone else mentioned to compare. Has anyone else done this comparison? My next trick is to find out how to get the Poloroid Digital Pallete we have to use these long exposure films, if possible.
Thanks to all who responded to my cry for help recently re: what I mistakenly was calling moire patterns on images scanned using my Agfa Arcus II scanners.
Turns out, as was recently mentioned in a posting by Tina Calvaho (sorry if I got that wrong Tina), that the the effect is attributed to "Newton Rings" by those in the biz. The problem shows up when scanning negs which don't have very much detail.
The primary culprit in my case was a misalignment of the transparency module (ie the lid was on crooked). The effect was most prominent on the scanner with the greatest misalignment. This is not something which you can adjust unfortunately (holes in the chasis which accept the mounting brackets were slightly off), but Agfa was quick to send me another unit with it's head on straight.
None of my units are "perfect", but I'm told that something called "Newton Ring Spray" or matte spray solves any remaining problem. I'm hesitant to spray something on the glass, but if it's removable, I'll likely give it a try.
Cheers,
**************************************** Don Steele Steele-at-KRDC.INT.Alcan.Ca Alcan International Kingston Research and Development Centre (613) 541 - 2145 ****************************************
} comparison? My next trick is to find out how to get the Poloroid Digital } Pallete we have to use these long exposure films, if possible.
Rick brings up a point that should be considered when evaluating a scanner. Not only should you consider the pixel depth (8, 10 or 12 bits), but you should also consider the optical density range of the scanner. I don't think the recent thread on scanners touched on this.
The digitization depth basically indicates the *output* range of the scanner. A 10 bit scanner will output ~1000 gray levels. The optical density range, though, indicates the *input* range of your scanner. Optical density (OD) is the log(10) of the fraction of transmitted light. Since most scanners start at roughly an OD of 0, a scanner with an OD range of 2 will be able to "see" down to about 1% light transmission. An OD range of 3 provides you with a range of 1000 in the light transmission. With the same number of bits, each step will be 10 times coarser. You will, however, be able to pull information from those dark regions of the negative. Consider what kind of negatives you will be scanning. If they are relatively flat, an OD range of 2 may be sufficient. If you have high contrast negatives, check whether you need the extra input range.
The Umax scanner I've used has an optical density range of ~2.0. Agfa indicates that their Arcus II (US$1500-2000) has a density range of 3.0 (0.2 - 3.2). Their DuoScan ($4000-4500) has an OD range of 3.3 (0.2 - 3.5?). Polaroid's Sprintscan 45 apparently has an OD range of 3.4 with a $10k list price (ouch!).
So... think about your OD needs as well as pixel density, digitization bits, and price!
Cheers, Henk
Henk Colijn colijn.1-at-osu.edu OSU Campus Electron Optics Facility ------------------------------------------------------------------- Prosperity is the blessing of the Old Testament; Adversity is the blessing of the New. Francis Bacon, "Of Adversity."
Other than toluidine blue, as you mentioned, we also use a variation on the hematoxylin theme for our epon sections. We use Heidenhain hematoxylin-iron which comes in two solutions (I have the recipes if you want to make your own) - an iron alum solution and a hematoxylin solution. Slides are preheated and stained on a hot plate that is maintained between 80 and 90 degrees C. Coat the sections with the iron alum solution for 2-10 min depending on the tissue type, thickness, etc. Rinse VERY WELL with distilled water. Repeat the process on the hot plate with the hematoxylin staining solution using the same time interval as the iron alum. Because the "staining" only appears at this step, you may need to test a few slides to determine ideal timing. After rinsing well again with distilled water, flood the slides on the hot plate with TAP water to differentiate and let sit for 3 min. Rinse briefly in distilled water and dry. Prestaining like this allows the slides to then go through autoradiography without interfering with the emulsion and has worked for us irregardless of the fixative used.
Pat Hales McGill University Dept. of Anatomy & Cell Biology hales-at-hippo.medcor.mcgill.ca
I am looking for some one who as any information on something called antistatic "Denkil SEM" (Hodogaya Chemical Co. Ltd.). It was cited in a short article in J. Electron Microsc, Vol 28, No. 4 P 312-313.
Hoping to spark some ideas I have included the article, sic. (I apologize for any inconvience for members who are charged by file size basis, but its quite short:). Please, excuse any typos as I am not a professional typist, but I haven't altered any wording. Sorry I haven'y included the figures (but they do look impressive enough to bother trying to follow up)
----+----+----+----+----+----+----+----
J. Electron Microscopy, 1979, Vol. 28, No.4, 312-313.
Scanning Electron Microscopy of Lily Pollen Grains treated with Antistatic Solution
Masaru Katoh
(Nissei Sangyo Co. Ltd.)
Pollen grains are generally observed by SEM after drying in air and coating with metal. in some special pollens or in studies with higher resolution and magnification, however, routine procedures for preparation of general biological materials may be necessary.
In this letter, effects of acohol-soluble antistatic agent (1) upon electroconductivity of the specimen of lily pollen grains are introduced. in this method, specimens do not need troublesome metal coating and preserve much more natural shapes.
Figure 1a shows pollen grains of lily simply dispersed on a small piece of adhesive tape without following procedure of metal coating. in this micrograph, a trouble of charging up of the specimen is recognized. figure 1b is a micrograph from the same area as shown in Fig. 1a, but was taken after the treatment that the specimen drawn out from the microscope was immersed in several drops of 3% alcohol solution of the antistatic "Denkil-SEM" (Hodogaya Chemical Co. Ltd., Japan), and throughly dried in air. by this treatment, charging up of the specimen was completely eliminated. Another important effect of this method is that the pollen grains are expanded and the net pattern of the surface structure is restored. This may be a big advantage of this method keeping the pollen grains closer to the natural shape. This effect is probably due to a strong affinity of the antistatic agent with water restoring the orginal shapes of the pollen grains. Effective water may be derived from the dissolving component in alcohol and humid atmosphere. the absorbed water is probably retained firmly in the pollen grains even after exposuring to electron beam in a high vacuum.
Such an electroconductive treatment using water-retainable antistatic agent is considered very useful not only for prevention of charging up of specimens, but also, to some extent, for keeping some specimens moist even in a high vacuum.
Further examples of application of this antistatic agent will be reported in the near future together with many other interesting results.
Reference:
(1) Katoh, M. : J. Electron Microsc., 28, 51 (1979)
-----+------+------+-------+-------+-------
That's it thats the entire paper. Does anyone out there have any ideas to point us in the right direction?
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513-529-5712 Fax: 513-529-4243 E-mail: edelmare-at-muohio.edu
I've noticed a great deal of discussion regarding films for slides, and given how important this topic is to microscopists, I thought I'd add my experience to the list. My original film was also the MP 5360. The film works well, is of high resolution, and exposure times of one second or less using a MP-4 copy stand are common. You can process it in your own lab. The drawback, in my opinion, is a yellowish-brown hugh in the background. I am told this can be cured with fresh bulbs in the MP4 copy stand, and that they should be replaced "routinely", but I really do not want to be bothered with that responsibility and expense. I have also used the polaroid instant slide film, but I find the resolution unacceptable, and the slides I have made and archived do not seem to hold up to the test of time. Our current solution? We use Kodak Ektachrome 64T ("T" for tungsten lighting), EPY 135-36 film. The resolution is top shelf, and exposure times are on the order of 1 second or less. Standard E-6 processing at the local film processing stand does a nice job, so you don't even have to get your hands wet. On a 35mm camera set up on the MP4 copy stand, we use a film speed of "50", and f-stop of 2.8, and automatic exposure. I do not know the specifics about the lens which we use, but the distance to the camera is from about 8 inches to 2.5 feet, depending on the enlargement desired. With this film you will also experience a brownish background. Using a SLIGHTLY blue filter over the camera lens (available at the camera shop), a pleasing grey background can be gained. As a darker blue lens is used, the slides will become increasingly blue, but I for one prefer blue over brown.
Hope this helps,
Doug Keene Shriners Hospital for Children Portland, Oregon ---------------------- Doug Keene DRK-at-shcc.org
I recently purchased the Tektronics Phaser 550 colour laserwriter with extended features (1200 dpi) at great cost - ~$12K (OZ$). It arrived last week to great joy but test prints showed a regular fine horizontal line repeated at a spacing of 26.1mm when printing at premium grade. The spacing is 35mm at all other grades. The line is ~0.1-2 mm in width. It occurs in all images, ie colour, B/W and on test colour stripe images, ie it is independant of image type or source.
Complaint led to a rapid visit by the service engineer who confirmed it as a problem and that a second complaint had been lodged from a similar new installation several days earlier.
The Company response is now - that's laserprinters! IE the Tektronics Phaser 550 will always produce this regular and quality destroying line. I agree it is not strong but once noticed it catches the eye.
My query is:
has anyone else observed this regular lining?
Has anyone got a Phaser 550 which does not produce the lines?
I feel that this is totally unacceptable, needless to say these lines were not evident in sample prints, but the overall quality is good and the media print pricing is also good.
Note I am aware of dye-sub etc, I was after a unit with a lower media unit cost.
Thank you
Brendon J. Griffin Centre for Microscopy and Microanalysis The University of Western Australia Nedlands, WA, AUSTRALIA 6907 ph 61-9-380-2739 fax 61-9-380-1087
*Please note change to email address: bjg-at-cyllene.uwa.edu.au
Dear Andreas, The simple answer is: 1.) Use carbon evapoative coating when you want to do an EDX analysis of the ceramic. The thickness of carbon when a polished brass disk beside your sample just turns blue is 25 nm and is adequate for most samples. 2.) Use sputtered gold or sputtered 60%gold - 40%palladium alloy when imaging is your goal. Images are better when the sample is gold sputtered. A 10 nm coat is good. Your sputter coater should have a time vs. coating thickness chart. You wrote: } we recently got a new SEM. } } What we need now is some advice for coating ceramic samples } When to use Carbon, Gold, thickness etc. } } Is there any FAQ about this available? } } Any help highy appreciated. Good luck with your new SEM. Regards, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
Job vcancy for PhD student / Research Cooperation within area of biomimetic chemistry
We are looking for a PhD student to work on a project focusing on " Chemical reactions at microstructured surfaces ". The project is in cooperation with an industrial partner and the Central German Research Agency ( Leitz Electron Optics , LEO ) and focuses on the - chemical surface modification by microprinting techniques as published by Whitesites et. al. - study of surface reactions (for example formation of inorganic phases at patterned surfaces) within patterned surfaces in order to create spatially defined product areas. The work follows the ideas of biomimetic reactions at organic (polymeric) surface layers.
The structural part of the work focuses on the structural characterisation of surfaces and the reaction products formed at the surface. This should be done by microscopic methods inlcuding high resolution field-emisson scanning electron microscopy as well as Energy spectroscopic imaging and EELS analysis.
Experience in the surface modification by micropatterning is already available.
The project lasts till 31.12.1999
A cooperation with groups engaged in surface reactions, biomimetical reactions could also be possible. (Exchange of PhD students within the frame of this project).
For further information contact :
Dr. H.-G. Braun Institute of Polymer Research Dresden Microstructure Groups D-01069 Dresden Hohe Str. 6 Germany
We have experienced the same with a Lexmark Optra R at the higher dpi settings. Company says that it will always be a problem because the manufacturing process allows a rather high tolerance in the production of the gearing leading to chatter. They have given us two other machines which show the same problems. Needless to say we are quite dissatisfied with the product.
At 11:37 AM 3/20/97 +0800, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 352-392-1295 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
Hello Everyone, There has been a tremendous response to my plea for information about the DPM 2468. At the request of a couple of you, I am posting my findings to the list. Here is basically what I have learned from you folks, Kodak, and a couple of dealers with whom I have spoken.
1. Kodak is still making the DPM 2468. They won't produce the film, however, until they have an order for it.
2. Kodak will not sell this film in quantities less than 50 100ft. rolls. This makes dealers reluctant to stock it.
3. Neelima Shah informed me that Mid City Camera in Philadelphia carried this film and I was able to order 2 rolls from them. However, they are not sure that they will continue to stock it and are going to call me later this week to let me know what they decide. The 2 rolls they sold to me were the last they had in stock.
Now for the good news: Several of you have informed me of another film which seems to be very similar to this one in ease of use, exposure times and development. It is called Direct MP Film 5360 and is available from Ted Pella. You can find it on page 186 of their catalog. Those of you who have used this film seem to be pleased with the product. I would like to thank all of you who responded to my post on this listserver and for the information you shared with me. I attempted to answer each of you individually; but because the response became so great, I will have to thank you as a group. Once again you were there and willing to help. I am not affiliated with any of the organizations who make or sell any of the products discussed here. Again, thanks to all who shared. Sandra
Sandra F. Zane, EM Tech. sfzane-at-email.uncc.edu Dept. of Biology, UNCC Ph.(704)547-4051 9201 University City Blvd. Fax (704)547-3128 Charlotte, NC 28223
I am currently trying to switch from Epon embedding media to one which polymerizes by UV at lower temperatures. In order to orient my tissue properly I would like to continue using flat embedding molds. My problem is in making these molds "airtight" in order to prevent shrinkage/evaporation while it polymerizes. Has anyone had this problem or found a solution to it?
Pat Hales McGill University Dept. of Anatomy & Cell Biology hales-at-hippo.medcor.mcgill.ca
I have a Voyager EDX system. I want to perform a standardless semi-quant analysis on a solid solution. The solid solution is a combination of Barium, Strontium and Calcium carbonates, (BaCO3, SrCO3 & CaCO3). Can I have the Voyager look at the Ba, Sr & Ca and tell it that these exist as carbonates and have it calculate the weight percentage of each from me entering in the stoichiometry of the solution? Are there other suggestions for me to go about measuring the weight percentages of Barium carbonate, strontium carbonate and calcium carbonate in a solid solution?
} Date: Thu, 20 Mar 1997 16:32:07 } To: pat hales {hales-at-medcor.mcgill.ca} } From: Scott Whittaker {sdw-at-biotech.ufl.edu} } Subject: Re: UV Embedding in Flat Molds } } } } A procedure we use around here successfully is a piece of cleared film ( or aclar) punched with a hole puncher and placed on a pre-polymerized resin surface in a centrifuge tube. Just drop in the film, add tissue, add the resin, and polymerize as you would normally. } After curing, remove the tube and break the sample at the disk level. The resin doesn't stick to the film (or aclar) and you are left with the tissue at the surface of the block and ready to section in any orientation. Good luck } } } } } } } At 02:50 PM 3/20/97 -0800, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 352-392-1295 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
The Pacific Northwest Electron Microscopy Society wishes to extend a warm welcome to all who wish to attend the Spring '97 Meeting at the V.A.Medical Center in Portland, Oregon Friday May 2nd, 1997. There are still a few spots for the up and coming young investigators/ graduate students to present papers on their current work to an enthusiastic Northwest audience of colleagues. Please send an e-mail to Charlie Meshul, Program chairman at meshulc-at-ohsu.edu with your topic if you wish to added on. Alternatively send e-mail to Bob Kayton kayton-at-ohsu.edu or Bob Mixon mixonr-at-osu.edu
Has anyone hired a commercial enterprise to conjugate their primary antibody to a gold particulate? We have a couple of mouse monoclonals that we would like to simultaneously localize in tissue, but do not have the time or experience to do the conjugations ourselves.
Many thanks,
Doug Keene Associate Investigator Shriners Hospital for Children
Greetings, For flat embedding under uv light without air, we use flat bottom capsules. These are made by TAAB in UK and distributed by several USA em suppliers. If you have really big samples, I beieve the little plastic widgets that em grids come in will work and these too are available from suppliers (EMS at least, and no doubt others too). Hope this helps,
I have hundreds of 35mm color negs which I wish to make proof sheets of. I do not want to do anything with those images other than print proof sheets. Can anyone suggest a good scanner for this job.
I have a PhD student from Brazil who is studying Catolaccus grandis. He places a female on top of a beeswax cell which contains boll weevil larva. The female deposits an egg on the larva and 2 weeks later, it matures and burrows its way out of the beeswax creating a hole. He would like to study and measure these holes.
However, from past experience I know that beeswax melts when placed in my Au/Pd sputter coater. Is there any way to coat the beeswax and examine them with an SEM? Or should he try another route?
Thank you in advance,
Ginger Baker EM Lab Manager Dept. Anatomy, Pathology, and Pharmacology 250 Veterinary Medicine Oklahoma State University Stillwater, OK 74078 (405) 744-6765 FAX: (405) 744-5275 Email: lizard-at-okway.okstate.edu
Regarding your question about flat embedding samples in media which polymerize under U.V.. We have used both LR White and Lowicryl resins successfully for flat embedding. We use Beem flat embedding molds, which are quite stiff and seem to contain less free oxygen than standard molds. Our trick to exclude atmospheric oxygen is to use the plastic pipette tip box that "Multi-Flex" tips are packaged in. It is a three part box which consists of a UV light penetrable top, which fits over a multi-holed mid section (normally supporting the tips) which fits over a lower compartment. I collected a bunch of these from another lab, but I do not know where to get them. We fill the lower compartment with dry ice (solid CO2), and drill a couple of holes in the top (letting CO2 gas escape as the solid sublimes). The samples in the beem flat embedding molds are placed on the perforated "shelf" above the CO2 chamber and exposed to a constantly replenished environment of gaseous CO2, excluding incoming oxygen. We put the whole thing, including a UV light, in the -20 portion of our freezer. The polymerization proceeds fully enough by the time the dry ice is gone to prevent oxygen inhibition. You might also try using a "heat sink" type of mold. Ted Pella sells one using a teflon mold which slides over aluminum ( I have no commercial interest in Pella).
You didn't mention what kind of UV-polymerizing resin you'll be using, but if it is Lowicryl or a similar methacrylate, here's how to do it:
1. Use polyethylene flat embedding molds. These molds are made from the same type of material as BEEM (R) capsules. The polyethylene molds are transparent to the UV light, making the polymerization much more even. Get back to me if you can't find a vendor for the polyethylene molds, I can't remember at this moment who sells them -- Ted Pella, I think...but I can look it up for you.
2. Place the mold on a piece of cardboard that is wrapped in aluminum foil. The cardboard should be larger than the mold. This makes moving and transferring the mold much easier and less messy (see below).
3. Overfill all of the cavities in the mold to give a "positive meniscus" at each position. If some of the cavities don't have specimens in them, go ahead and fill the cavities anyway.
4. Cut a piece of Parafilm (R) that is slightly larger than the mold. Then, beginning at one end of the mold, gently lay the Parafilm down on top of the mold. As you do this, the excess resin will run out of the mold cavities and get trapped under the Parafilm. Gradually lower the Parafilm down toward the other end of the mold, allowing the resin to fill the space between the mold and the Parafilm. This seals the complete top of the mold. The resin won't dissolve the Parafilm, but it will make it soft.
5. You can use tissues to absorb the excess resin as it runs down the outside of the mold. (**CAUTION:** Be sure to wear gloves at this step, and at all steps when working with methacrylate resins -- or any resin, for that matter!).
6. Transfer the filled molds to your polymerization apparatus. I don't know what you will be using, but I make polymerization chambers out of cardboard boxes. Just make two cutouts: one on the top for the UV lamp, and one on the side for a door. **To make the polymerization even, cover all of the inner surfaces of the box with aluminum foil.**
7. **NOTE**: You can also use Thermanox (plastic) cover slips instead of Parafilm in Step #4. Just lay the coverslips on top of the cavities, letting the excess resin run out along the edges.
8. Let everything equilibrate at low temperature for 15-20 minutes, turn on the UV lamp, and you're on your way to polymerized resin!
9. **NOTE**: If you see "bubbles" or vortex-like "swirls" in the polymerized blocks, this is usually a sign that the polymerization was too rapid. Increase the distance between the lamp and the mold to cut down on the UV intensity. If this isn't possible, place a piece of frosted glass between the lamp and the molds.
I've never asked anyone to conjugate a monoclonal Ab directly to colloidal gold, mainly because of the added cost (a similar concern of yours, I take it!).
Are you up for a slightly different approach, which you can do with commercially available reagents? If so, you could do the following:
1. On one side of the grid (maybe a fairly high hexagonal mesh, uncoated nickel grid?) incubate with one of your mouse monoclonals.
2. Follow with biotinylated goat anti-mouse Ig-whatever (is it an IgG?).
3. Follow with streptavidin-gold (available in many different sizes).
4. Flip the grid over and repeat the labeling with the second monoclonal. Only this time, use a different size of streptavidin-gold.
5. Double sided labeling works quite nicely for co-localizations, and it eliminates a lot of cross-reactivity problems you'd have to fight if you worked on only one surface of the section.
6. One thing to watch out for: If you add normal goat serum and maybe a little TWEEN or other additives to the reagents, the grids will tend to sink if they're incubated on drops of the reagents. Then you're labeling *both* sides with the *same* reagents. This, of course, is to be avoided! You have to keep the sides separated.
The best way to avoid this problem is to anchor the grid by its edge on a piece of double-stick tape on a microscope slide. You can then use a micropipette to deliver *very small* volumes (maybe 2-3 microliters) of each reagent. To change reagents or to wash, simply add the reagent to one edge of the grid and wick it off with filter paper from the opposite edge.
Following the first gold incubation you can remove the grid and wash well in a gentle stream or w/ repeated drops of buffer and finally water. Allow to dry, flip the grid over, stick it down to the tape and do the second run-through.
An added bonus of this procedure is a fairly good signal amplification. If you conjugated your primary directly to the gold, you'd only have a one-step reaction with minimal gold particles (probably 1) binding at each site. Would that be enough to detect above background?
Unless, of course, you *want* the most specific localization possible...In that case, forget everything I've just written!
I've never asked anyone to conjugate a monoclonal Ab directly to colloidal gold, mainly because of the added cost (a similar concern of yours, I take it!).
Are you up for a slightly different approach, which you can do with commercially available reagents? If so, you could do the following:
1. On one side of the grid (maybe a fairly high hexagonal mesh, uncoated nickel grid?) incubate with one of your mouse monoclonals.
2. Follow with biotinylated goat anti-mouse Ig-whatever (is it an IgG?).
3. Follow with streptavidin-gold (available in many different sizes).
4. Flip the grid over and repeat the labeling with the second monoclonal. Only this time, use a different size of streptavidin-gold.
5. Double sided labeling works quite nicely for co-localizations, and it eliminates a lot of cross-reactivity problems you'd have to fight if you worked on only one surface of the section.
6. One thing to watch out for: If you add normal goat serum and maybe a little TWEEN or other additives to the reagents, the grids will tend to sink if they're incubated on drops of the reagents. Then you're labeling *both* sides with the *same* reagents. This, of course, is to be avoided! You have to keep the sides separated.
The best way to avoid this problem is to anchor the grid by its edge on a piece of double-stick tape on a microscope slide. You can then use a micropipette to deliver *very small* volumes (maybe 2-3 microliters) of each reagent. To change reagents or to wash, simply add the reagent to one edge of the grid and wick it off with filter paper from the opposite edge.
Following the first gold incubation you can remove the grid and wash well in a gentle stream or w/ repeated drops of buffer and finally water. Allow to dry, flip the grid over, stick it down to the tape and do the second run-through.
An added bonus of this procedure is a fairly good signal amplification. If you conjugated your primary directly to the gold, you'd only have a one-step reaction with minimal gold particles (probably 1) binding at each site. Would that be enough to detect above background?
Unless, of course, you *want* the most specific localization possible...In that case, forget everything I've just written!
Pat Hale wrote: ================================================== I am currently trying to switch from Epon embedding media to one which polymerizes by UV at lower temperatures. In order to orient my tissue properly I would like to continue using flat embedding molds. My problem is in making these molds "airtight" in order to prevent shrinkage/evaporation while it polymerizes. Has anyone had this problem or found a solution to it? ================================================== There is an alternative to the rigid polyethylene based (e. g. BEEM manufactured) flat embedding molds, and that is the flexible and reusable UV transparent silicone flat embedding molds. These silicone molds work just fine and seem to have a bit higher transparency for the UV. One would follow the method described by Bob Chiovetti, that is, to overfill the cavities but instead of using Parafilm, just place the bottom of another one of the UV transparent silicone molds on top. Capillary action quickly seals out any entrapped air, creating the desired oxygen free environment.
UV transparent embedding molds come in varous sizes and shapes and are shown on the SPI website given below. Similar, but not idential molds are also offered by Ladd and possibly others.
Disclaimer: SPI Supplies manufactures UV transparent silicone embedding molds and has a vested interest in seeing more persons using them!
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
The Polaroid Sprint Scan 35LE will scan your negatives in at resolutions up to 1950 dpi -at- 10 bits per channel with an optical density of 3.0 in under 60 seconds. It will scan 35 mm negatives or slides(mounted or unmounted)as well as the SuperSize format. It has an option called the PathScan Enabler that allow you to scan a standard glass slide at the referenced resolution and color depth. It has a list price of $995US.
John D. Warren Area Sales Manager Digital Products Polaroid Corporation
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have hundreds of 35mm color negs which I wish to make proof sheets of. I do not want to do anything with those images other than print proof sheets. Can anyone suggest a good scanner for this job.
POSITION AVAILABLE!!!! At the Institute for Neurobiology, University of Amsterdam, The Netherlands a post-doc position is available, for 3 years. The project concerns plasticity in brain cells, in relation to epilepsy and hormonal influences. It will make use of (fast) calcium imaging techniques with confocal microscopy, in combination with patch clamp recording. We are looking for someone with experience in confocal microscopy, preferably in brain tissue. The position is available as of now. For more information: wadman-at-bio.uva.nl or joels-at-bio.uva.nl or call 31205257641 / 31205257626.
----------------- P.C.Diegenbach dept. Biology, University of Amsterdam Kruislaan 320 1098 SM Amsterdam, Netherlands phone (31) 20 5257631 fax (31) 20 5257709 The following email address is mangled to prevent automated unsolicited junk mail. Replace the '_AT_' with an '-at-': email paulcd_AT_bio.uva.nl -----------------
Pat: I use UV polymerization of LR white or Unicryl at 4C for immunolabelling studies. I use small aluminum weighing dish layered with a sheet of Aclar film at the bottom. The embedding and tissue are placed on top of this film. Cut a sheet of closely fit Aclar film to place over it. All the weighing dishes are placed in a stainless tray which is sealed with Saran Wrap. The aluminum will reflect the Uv and provide good even polymerization. The embedding medium between the sheet will polymerize while some excess media flow over and under the sheet will not. The Aclar sheets are easily peeled off from the polymerized material. Sheets of embedding material can be scanned under LM to chose for ideal orientationed sample. Hope this will help you.
Licy Yin Microscopy Center U.Mass , Amherst, MA 01003t MiclOn Thu, 20 Mar 1997, pat hales wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I am currently trying to switch from Epon embedding media to one which } polymerizes by UV at lower temperatures. In order to orient my tissue } properly I would like to continue using flat embedding molds. My problem is } in making these molds "airtight" in order to prevent shrinkage/evaporation } while it polymerizes. Has anyone had this problem or found a solution to it? } } Pat Hales } McGill University } Dept. of Anatomy & Cell Biology } hales-at-hippo.medcor.mcgill.ca }
We have had superb results with Fuji's Velvia. It is a daylight film, so you will need blue filtration when used with tungsten halogen sources (82B)and it is slow (50ASA) but it produces exquisite color rendition and depth. It is also a standard E-6, for easy development. Good luck... and if you take an especially beautiful shot which you would like to share, let us know. We are always looking for interesting images for our short courses (with credit, of course).
Barbara Foster Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108 (413)746-6931 fax: (413)746-9311 email:mme-at-map.com
Pat Hales asks about airtight seals for acrylic embedding medium.
Unicryl, an acrylic embedding medium, advertises that excluding oxygen during UV polym is not necessary. Unicryl is available from a number of vendors. (I have NO interest in the commercial uses of Unicryl). Using an acrylic which bypasses the entire problem of oxygen interference might be a good solution. Bye, Hildy
MIDWEST MICROSCOPY AND MICROANALYSIS SOCIETY affiliate of the Microscopy Society of America and the Microbeam Analysis Society
presents
WORKSHOP AND SYMPOSIUM ON IN SITU HYBRIDIZATION
The program has been arranged by and will be held at Madison Area Technical College (MATC), WI., Friday, April 11, 1996 from 8:00 - 5:00 p.m. This meeting will consist of two concurrently held workshops and an afternoon related symposium. The general emphasis will be in situ hybridization. The two morning workshops will be on detection of hybridization with radioactive and non radioactive probes.
The meeting has several purposes:
1. To draw the attention of the scientific community to emerging developments in the practical and basic research aspects of excitingnew fields.
2. To bring people together from diverse disciplines in order to discuss how innovative techniques will be relevant to the future direction of microscopy and microbeam analysis.
WORKSHOPS WILL BE HELD IN THE MORNING FROM 8:30 -12:30.
1. Radioactive in situ hybridization Patricia L. Allen/Dr. Lyn Thet Department of Pulmonary Medicine, School of Medicine UW-Madison
2. Nonradioactive in situ hybridization Boehringer Mannheim, Indianapolis, IN
You may choose to attend one workshop from above. Registration information is enclosed. Register early because we will have to limit the number of participants.
AFTERNOON SYMPOSIUM SCHEDULE 1:30 - 5:00 p.m.
Patricia Thomas, M.D. Department of Pathology, School of Medicine, University of Iowa, Iowa City, IA, "Diagnostic applications of fluorescence in situ hybridization"
Kathrine Staskus, Ph.D. Department of Microbiology, University of Minnesota, Minneapolis, MN, "In situ hybridization detection of Simian Immunodeficiency virus (SIV) proteins and nucleic acids"
Gary Lyons, Ph.D. Department of Anatomy and Cellular and Molecular Biology Program, School of Medicine, University of Wisconsin, Madison, "In situ hybridization studies of murine embryonic skeletal muscle development"
Alan Smith, PhD. Department of Horticulture, College of Agriculture, University of Minnesota, Minneapolis, MN, "Localization of floral specific gene products during pollen development"
Gwen V. Childs, Ph.D. Vice Chair, Department of Anatomy and Neuroscience, Program Director. Cell Biology Graduate Program, University of Texas Medical Branch, Galveston, TX, "Regulation of multipotential cells in the pituitary by neuroendrocrine peptide:an in situ hybridization and immunocytochemical study"
Kevin Roth, M.D., Ph.D Department of Pathology, Washington University, St. Louis, MO. "Application of tyromide-signal amplification to in situ and immunocytochemistry"
WORKSHOP AND SYMPOSIUM ON IN SITU HYBRIDIZATION
GENERAL INFORMATION SHEET
DATE: Friday, April 11, 1997
LOCATION: Madison Area Technical College (Truax Campus) Electron Microscopy Program (Check-in, Rm375A) 3550 Anderson St., Madison, WI. 53704
Workshops and Symposium locations will be provided at the Check-in room (Rm375A)
DIRECTIONS PARKING: A map is provided on request. Please park in the large lot on the side of the campus building.
MEALS: Refreshments will be provided in the morning and for afternoon break. Lunch is a sandwich buffet, and the cost is $4.00/person. Following the afternoon symposium there will be a wine, soft drink, and cheese reception provided at no cost to participants. An Italian dinner buffet is being organized for after the wine and cheese reception. The cost is $7.00/person.
REGISTRATION: If you plan to attend either one of the workshops or the symposium you must register by phone or EMAIL to one of the contacts listed below. Registration is free to all Society members, student, regular, and corporate. If you are not a member of the Society, registration cost will be the price of membership. (Regular = $10, Student = $5). When you register you must provide us with the information listed below. Name tags and Fees for registration and meals will be collected at the Check-in site.
DEADLINE DATE FOR REGISTRATION IS APRIL 8.
1. Name 2. Affiliation 3. Phone # 4. Choice of one Workshop 5. Are you attending the Afternoon Symposium? 6. Do you want to participate in the Lunch or Dinner?
REGISTRATION CONTACTS:
James P. DiOrio (847) 270-4676 Fax: (847) 270-4414 EMAIL: diorio-at-baxter.com
Joanne M. Crudele (847) 734-3712 Fax: (847) 734-3686 EMAIL: Joanne.Crudele-at-Unilever. Com
If you need information on hotels, contact Michael Kostrna (608)246-6762 or Kenneth Muse (608)243-4309.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello all, } } I have a PhD student from Brazil who is studying Catolaccus } grandis. He places a female on top of a beeswax cell which contains } boll weevil larva. The female deposits an egg on the larva and 2 } weeks later, it matures and burrows its way out of the beeswax } creating a hole. He would like to study and measure these holes. } } However, from past experience I know that beeswax melts when placed } in my Au/Pd sputter coater. Is there any way to coat the beeswax and } examine them with an SEM? Or should he try another route? } } Thank you in advance,
} Email: lizard-at-okway.okstate.edu } Do you have cooling system on the sputter coater ? If your answer is yes, let the beeswas sit on the cooling stage for 20 min then coat it for 5-10 sec and pause for about 20 sec. Repeat it for 3-4 cycles. My experience for fibric material the above tricks would work fine. Also you may lower your sample far from the gold target to avoid heat.
Hope this will help.
*********************************************** * Ming H. Chen, PhD * * Medicine/Dentistry Electron Microscopy Unit * * University Of Alberta. * * Edmonton, Alberta, Canada * * * * Visit My Page At: * * http://www.ualberta.ca/~mingchen * ***********************************************
With a Special Lecture and Optional Demonstration Workshop on in situ Hybridization on Sunday, April 27 presented by Dr. L=E1szl=F3 K=F6m=FCves of UC San Francisco
Business meeting for CSU delegates, Friday, April 25, 1997 at 4:00 PM in Duncan Hall Rm. 249
Proceedings will be published in Microscopy Research and Technique
The Colloquium provides a forum for the exchange of research results and experiences in all fields of light and electron microscopy in the biological, geological, and materials sciences. Participants include students and scientists from academia and industry.
Both platform and poster presentations are invited. Student presentations are strongly encouraged.
Registration Fees $35 Regular, $10 Student, $50 Vendor by March 15; $25 Optional Workshop
Late Registration $45 Regular, $10 Student, $60 Vendor, after March 15; $25 Optional Workshop
Make checks payable to San Jose State University Foundation
Send to: Dr. David K. Bruck CSU Microscopy Colloquium Department of Biological Sciences San Jose State University San Jose, CA 95192-0100
=46or additional information, flyers, and abstract forms, contact David Bruck at phone: (408) 924-4837 or fax (408) 924-4840 bruck-at-biomail.sjsu.edu
Program
=46riday, April 25
4:00 PM Business meeting for CSU delegates in Duncan Hall Rm. 249, San Jose State University
Saturday, April 26
8:00 AM Registration, coffee, and breakfast snacks in lobby of Duncan Hall, San Jose State University 9:00 AM Opening remarks in Duncan Hall Rm. 135 9:15 AM Platform presentations 10:30 AM Coffee break 11:00 AM Platform presentations 12:00 PM Buffet lunch 1:30 PM Platform presentations 2:30 PM Coffee break 3:00 PM Platform presentations 4:00 PM Lecture on in situ hybridization by Dr. Laszlo Komuves 5:00 PM Closing Remarks and Poster session in Duncan Hall Rm. 250 6:30 PM Banquet
Sunday, April 27
9:00 AM Demonstration and workshop on in situ hybridization conducted by Dr. Laszlo Komuves in Duncan Hall Rm. 344
Housing
Best Western Inn 455 S. Second St. San Jose, CA 95113 phone (800) 528-1234 or (408) 298-3500 singles $52, doubles $62
Please make your own reservations for housing at one of the phone numbers above. Specify that you will be attending the Microscopy Colloquium to receive the conference discount. A map and other hotels in the vicinity will be included in the packet of information mailed to you following receipt of your registration information.
Transportation
=46ree parking on campus is available in the 7th St. Garage on the corner of 7th St. and San Salvador St. A permit will be mailed to you following receipt of your registration form.
Public transportation from the San Jose International Airport to downtown San Jose is limited to cabs or the Light Rail system. For the latter, a free shuttle bus (labeled Metro) runs every 10-15 minutes from the airport to the Metro Light Rail Station. For $1.10, the train runs downtown to the Paseo de San Antonio Station, about 3 blocks from the university and 2 blocks from the Best Western Inn.
Registration
Please complete the registration form below whether or not you will be presenting a paper or poster.
After March 15 $45 regular, $10 student, $60 vendor
_____________________________ Name
_____________________________ Address
_____________________________ City State Zip
_____________________________ Phone
_____________________________ e-mail
Workshop Registration (for an additional fee of $25) 0 Workshop
Note that the workshop's occurrence will depend on an adequate number of registrants.
Meal Preference 0 Chicken 0 Fish 0 Vegetarian
Presentation 0 Platform 0 Poster 0 None
Information and Mailing
Make registration checks payable to the San Jose State University =46oundation. Mail registration forms, abstract forms, and checks to
Dr. David K. Bruck CSU Microscopy Colloquium Department of Biological Sciences San Jose State University 1 Washington Square San Jose, CA 95192-0100
(408) 924-4837 bruck-at-biomail.sjsu.edu
Platform and Poster Presentations
Papers discussing any aspect of microscopy in cell, developmental, and structural biology or in the material or geological sciences are welcome. Microscopy techniques' papers are particularly encouraged.
Platform presentations (15 min.) will be held Saturday, April 26. Posters will be displayed all day Saturday, with an afternoon session designated for viewing with authors present.
An Abstract Guidelines form is included for those presenting papers or posters. The abstracts will be compiled into the "Proceedings of the Fifth Annual CSU Microscopy Colloquium" and will be published in Microscopy Research and Technique.
Dr. Steven Barlow EM Facility/Biology Department 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/EM_Facility
Carolina Workshop on: LIGHT MICROSCOPY FOR THE BIOMEDICAL SCIENCES June 1-6, 1997 University of North Carolina at Chapel Hill
Instructors: John J. Lemasters Edward D. Salmon Brian Herman ------------------------------------------------------------------- LIGHT MICROSCOPY FOR THE BIOMEDICAL SCIENCES is an introduction to applications of light microscopy. Students will have opportunities for extensive hands-on experience with state-of-the-art equipment for optical imaging, digital imaging processing, fluorescence microscopy and confocal microscopy guided by experienced academic and commercial staff. The course is divided into three major sections with lectures and laboratory exercises on: 1) geometric and wave optics of image formation, microscope alignment, phase contrast and reflection interference contrast microscopy; 2) video imaging, including contrast enhancement by analog and digital image processing, fluorescence microscopy, image detectors, fluorescent probes, ion imaging, and green fluorescent protein; and 3) laser scanning confocal microscopy emphasizing live cell imaging and 3-dimensional image reconstruction. Students are encouraged to bring their own specimens for analysis. A commercial staff representing leading microscopic manufacturers will make available for student use the latest and most advanced instrumentation for light microscopy, image detection and computerized image analysis.
Tuition is $950.
------------------------------------------------------------------- APPLICATION FORM Carolina Workshop on LIGHT MICROSCOPY FOR THE BIOMEDICAL SCIENCES
Position:
Address:
Telephone:
Fax:
Please return this form along with a brief letter describing your research interests and a curriculum vitae. Applicants should contact the program as soon as possible. Full consideration will be given to applications received by April 18, 1997.
Send application to: Dr. Wayne Litaker, Director of Workshops University of North Carolina at Chapel Hill Program in Molecular Biology & Biotechnology CB# 7100, 442 Taylor Hall Chapel Hill, North Carolina 27599-7100 Tel: (919) 966-1730 Fax: (919) 966-6821 e-mail: litaker-at-med.unc.edu ------------------------------------------------------------------- {End of Announcement}
I have one single question for the wealth of knowledge out there in the world of Microscopy....
On a Nikon Diaphot inverted with a universal Con.A Achr-Apl 1.4 condenser using a 10X objective with DIC do you have to oil the condenser????
Also how do you do the alignment properly, since noone in the lab I work knows... I think I have everything correct but when I twist polarizer by the objective lens I do not get the rainbow of colors or even the black background when the polarizers are ligned up....
Any Suggestions \\|// (o o) ~~~~~~~~~~~~oOOo~(_)~oOOo~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
{ { {This message is made of 100% recycled electrons} } }
Cheers ;o) :o) %o) Eric {Mesa Arizona} http://www.goodnet.com/~earosen (Note the tilde before earosen)
An alternative may be replication using Reprosil, a Polyvinyl Siloxane from Caulk. It makes a negative which can be transfered to a positive by using Spurr low viscosity embedding medium. Detail is there albeit a little softened.
I am a student now using Hitachi-8100 TEM to study submicrometer Cu-Zn alloy particals. The particals are mostly six-fold symmetry plates. When the beam focused on a partical at a high magnification, some dark stripes appear in the partical's bright-field image and change their shapes until a snow-flake-like pattern is formed. The pattern comprised of six center-crossed dark stripes. Select-area diffracton shows a single crystal diffracton pattern with six-fold symmetry. But dark-field image shows that one single diff spot is not coming from the whole partical but from ONE corresponding dark stripe.
My questions are: 1)Do anyone out there ever find similar phenomenon? 2)How can I know the temperature of the particals under the brightening of the electron beam? I think temperature is a reason for the formation of the snow-flake-like pattern.
I hope I have described my question clear enough for you to reply. Thanks!
Sun Haiping National Lab of Superhard Materials Jilin University Changchun 130023, P.R.China sunqa-at-mail.jlu.edu.cn
Aclar is a thin plastic sheet material suitable for cell culture and may be sectioned. A full description is in our catalogue on page L7. We distribute Aclar as does Ted Pella (Pelco) in the USA. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 300+ Links, MSDS ************************ http://www.proscitech.com.au
} Several of you have recommending using Aclar film in your UV polymerization } protocols. What is Aclar film, and where do you purchase it? } } Gary Radice, Associate Professor gradice-at-richmond.edu } Department of Biology 804-289-8107 (voice) } University of Richmond VA 23173 804-289-8233 (FAX) } }
Hello, I am having no luck attempting to stain the glycocalyx? of a species of sulfate-reducing bacteria using ConA-conjugated gold (5nm). Using 4%PF+0.1%Glute in Cac buffer fixation followed by LRW embedding I have tried 1:10 to 1:100 in P04 buffer pH 7.2. I have also tried ConA-gold in Tris-saline with and without 1mM CaCl2 and 1mM MgCl2 at pH 6.9,7.2 and 8.0. diluted 1:10-1:100. Next I tried taking a fresh culture and washing several times in Tris-saline followed by 1:10 ConA-gold,then fixing and embedding. My incubations have been around 1 hr at room temp. Nary a gold particle do Isee under any of the conditions mentioned. Apparently, using fluorescently labelled ConA these little buggers lightup like the Hale-Bopp comet, so Iam at a lost. Can anybody out there give me some suggestions. I also have some material that I fixed in 4% PF alone, embedded in LRW that I have not done anything yet with. Thanks in advance
Hank Adams Electron Microscopy Laboratory New Mexico State University Las Cruces, NM 88003
I have an American Optical Micro-Star Illuminator model 1872. It has 10XB&L WF eyepieces and a third place for camera/eyepiece. The primary voltage is 115 and secondary 3 to 6 volts using GE lamp 1974 It has no objectives, but spaces for 4. It has a large mechanical base. It is illuminated thru the lenses.
I know nothing else about the scope except it was used in a lab at Fairchild semiconductor in South.
Iam looking for some good quality/inexpensive objectives for amatuer use and any manuals/info on using this scope.
If you know anything of this scope or the objectives that I need, please e-mail me
Thanks so much for your time
David Chase dchase-at-gwi.net
ps I had researched this all once before but a fatal HD crash destroyed all my data.
Hello, I am having no luck attempting to stain the glycocalyx? of a species of sulfate-reducing bacteria using ConA-conjugated gold (5nm). Using 4%PF+0.1%Glute in Cac buffer fixation followed by LRW embedding I have tried 1:10 to 1:100 in P04 buffer pH 7.2. I have also tried ConA-gold in Tris-saline with and without 1mM CaCl2 and 1mM MgCl2 at pH 6.9,7.2 and 8.0. diluted 1:10-1:100. Next I tried taking a fresh culture and washing several times in Tris-saline followed by 1:10 ConA-gold. My incubations have been around 1 hr at room temp. Nary a gold particle do I see under any of the conditions mentioned. Apparently, using fluorescently labelled ConA these little buggers light up like the Hale-Bopp comet, so I am at a lost. Can anybody out there give me some suggestions. I also have some material fixed in 4% PF alone, embedded in LRW that I have not done anything yet with. Thanks in advance,
Hank Adams Electron Microscopy Laboratory New Mexico State University Las Cruces, NM 88003
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hank Adams writes: "Hello, I am having no luck attempting to stain the glycocalyx? of a species of sulfate-reducing bacteria using ConA-conjugated gold (5nm)." (trimmed).
Dear Hank, Your problem may lie in the ConA-conjugated gold. Often, these probes very seldom store well. If most of the protein has dissociated from the gold particles, you will have a large amount of very specific ConA binding but no gold to show you it is there. It is easy to test at the LM level by probing sections treated with ConA-god with anti-ConA and fluorescent antibody.
Once again, the best visualization method for lectins (re:ricin) may be to use antibodies instead of the directly conjugated gold probes. If you prefer to use ConA-gold then I recommend that you make your own. A slightly less satisfactory approach would be for you to centrifuge down your ConA-gold (be careful not to spin it so hard that is aggregates into the bottom of the tube) and resuspend it in fresh PBS. This will remove most of the free protein.
As an aside and in reference to a previous posting, streptavidin-gold also suffers from this storage problem. When using the probe at the LM level, with silver enhancement, or with multiple antibody layers the loss of labeling efficiency is not so noticeable. However, attempting to visualize biotinylated antibodies directly with streptavidin-gold will only work well for the first week after the probe has been made. For situations where there are large numbers of bound antibody, this effect may not be noticeable, but for situations where there are only low numbers of antigens, it is very obvious.
Making colloidal gold and coupling it to proteins or antibodies is easy and anyone with even limited experience can do it.
Check out my WWW "gold page" to see how easily it can be done. Be warned, if you want to do this you usually need large supplies of protein (a point anyone coupling antibodies should be aware of).
Paul Webster Center for Cell Imaging Yale School of Medicine http://info.med.yale.edu/cellimg
Message-Id: {199703230345.UAA07299-at-mailque.goodnet.com} Comments: Authenticated sender is {earosen-at-pop.goodnet.com}
You may also want to treat the sections with a very low concentration of sodium periodate which will destroy the section if you are not careful.. This will expose some of the bnding sites fo the Con-A.. or on the other hand the Lctins are too old, and the gold has fallen off of the lectin which occured to me also when I was using gold labeled lectins that were stored in the fridge........ So I got very scattered labeling or no labelling at all but when used in teh LM with Cy3 they light up like a chrsitmas tree....
lAlso you may want to try a lower comcentration of fixatives in Formaldehyde and Glut.
--------------------------------------------------------------------- --- The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com ---------------------------------------------------------------------- -.
Hello, I am having no luck attempting to stain the glycocalyx? of a species of sulfate-reducing bacteria using ConA-conjugated gold (5nm). Using 4%PF+0.1%Glute in Cac buffer fixation followed by LRW embedding I have tried 1:10 to 1:100 in P04 buffer pH 7.2. I have also tried ConA-gold in Tris-saline with and without 1mM CaCl2 and 1mM MgCl2 at pH 6.9,7.2 and 8.0. diluted 1:10-1:100. Next I tried taking a fresh culture and washing several times in Tris-saline followed by 1:10 ConA-gold. My incubations have been around 1 hr at room temp. Nary a gold particle do I see under any of the conditions mentioned. Apparently, using fluorescently labelled ConA these little buggers light up like the Hale-Bopp comet, so I am at a lost. Can anybody out there give me some suggestions. I also have some material fixed in 4% PF alone, embedded in LRW that I have not done anything yet with. Thanks in advance,
Hank Adams Electron Microscopy Laboratory New Mexico State University Las Cruces, NM 88003
Can any one tell me of a Microscope news group taylored toward the newbie into this hobby? Also, are there any good magazines on the market specializing in MIcroscopes and the Microscope hobby?
} Date: Sat, 22 Mar 1997 15:20:12 -0500 } From: H. ADAMS {hadams-at-nmsu.edu} } To: microscopy-at-Sparc5.Microscopy.Com } Subject: Lectin staining } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello, I am having no luck attempting to stain the glycocalyx? of a } species of sulfate-reducing bacteria using ConA-conjugated gold (5nm). } Using 4%PF+0.1%Glute in Cac buffer fixation followed by LRW embedding I } have tried 1:10 to 1:100 in P04 buffer pH 7.2. I have also tried ConA-gold } in Tris-saline with and without 1mM CaCl2 and 1mM MgCl2 at pH 6.9,7.2 } and 8.0. diluted 1:10-1:100. Next I tried taking a fresh culture and } washing several times in Tris-saline followed by 1:10 ConA-gold. My } incubations have been around 1 hr at room temp. Nary a gold particle do I } see under any of the conditions mentioned. Apparently, using fluorescently } labelled ConA these little buggers light up like the Hale-Bopp comet, so I } am at a lost. Can anybody out there give me some suggestions. I also have } some material fixed in 4% PF alone, embedded in LRW that I have not done } anything yet with. } Thanks in advance, } } Hank Adams } Electron Microscopy Laboratory } New Mexico State University } Las Cruces, NM 88003 } } http://www.nmsu.edu/Research/artsci/public_html/eml/ } } 505-6463600 } Since the glycocalyx is on the outside of the bacteria and you don't have to worry about sectioning to expose the conA to the glycocalyx, why don't you stain with con A-Au, wash thoroughly (centrifugations), and then embed normally?
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
} Date: 22 Mar 1997 18:39:10 -0500 } From: Paul Webster {paul.webster-at-Yale.edu} } To: "Microscopy -at-MSA.Microscopy.Com" {Microscopy-at-Sparc5.Microscopy.Com} } Subject: Re: Lectin Staining } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hank Adams writes: } "Hello, I am having no luck attempting to stain the glycocalyx? of a } species of sulfate-reducing bacteria using ConA-conjugated gold (5nm)." } (trimmed). } } Dear Hank, } Your problem may lie in the ConA-conjugated gold. Often, these probes very } seldom store well. If most of the protein has dissociated from the gold } particles, you will have a large amount of very specific ConA binding but no } gold to show you it is there. It is easy to test at the LM level by probing } sections treated with ConA-god with anti-ConA and fluorescent antibody. } } Once again, the best visualization method for lectins (re:ricin) may be to use } antibodies instead of the directly conjugated gold probes. If you prefer to use } ConA-gold then I recommend that you make your own. A slightly less satisfactory } approach would be for you to centrifuge down your ConA-gold (be careful not to } spin it so hard that is aggregates into the bottom of the tube) and resuspend it } in fresh PBS. This will remove most of the free protein. } } As an aside and in reference to a previous posting, streptavidin-gold also } suffers from this storage problem. When using the probe at the LM level, with } silver enhancement, or with multiple antibody layers the loss of labeling } efficiency is not so noticeable. However, attempting to visualize biotinylated } antibodies directly with streptavidin-gold will only work well for the first } week after the probe has been made. For situations where there are large } numbers of bound antibody, this effect may not be noticeable, but for situations } where there are only low numbers of antigens, it is very obvious. } } Making colloidal gold and coupling it to proteins or antibodies is easy and } anyone with even limited experience can do it. } } Check out my WWW "gold page" to see how easily it can be done. Be warned, if } you want to do this you usually need large supplies of protein (a point anyone } coupling antibodies should be aware of). } } Paul Webster } Center for Cell Imaging } Yale School of Medicine } http://info.med.yale.edu/cellimg } Also, if you're going to spin out your con A-Au from the uncongugated con A, keep the pH high (probably above 8-8.2, or whatever pH the original stuff was shipped in). Otherwise, you might have one big blob at the bottom of your tube. When we suspect unconjugated protein, we dilute the antibody-Au to the concentration we want to use with buffer at a pH above the pI (pH } 8), and then spin in a Beckman Airfuge 5 min at 30 lb (~100,000 g), discard the sup and resuspend the pellet in the same vol(pH 7) as we started with. Haven't tried con A; I assume it might work the same way???
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
} Date: Sat, 22 Mar 1997 20:46:17 -0800 } From: Eric {earosen-at-pop.goodnet.com} } To: microscopy-at-Sparc5.Microscopy.Com } Subject: Re: Lectin staining } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } You may also want to treat the sections with a very low concentration } of sodium periodate which will destroy the section if you are not } careful.. This will expose some of the bnding sites fo the Con-A.. or } on the other hand the Lctins are too old, and the gold has fallen off } of the lectin which occured to me also when I was using gold labeled } lectins that were stored in the fridge........ So I got very } scattered labeling or no labelling at all but when used in teh LM } with Cy3 they light up like a chrsitmas tree.... } } lAlso you may want to try a lower comcentration of fixatives in } Formaldehyde and Glut. } } } --------------------------------------------------------------------- } --- The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } ---------------------------------------------------------------------- } -. } } Hello, I am having no luck attempting to stain the glycocalyx? of a } species of sulfate-reducing bacteria using ConA-conjugated gold (5nm). } Using 4%PF+0.1%Glute in Cac buffer fixation followed by LRW embedding } I have tried 1:10 to 1:100 in P04 buffer pH 7.2. I have also tried } ConA-gold in Tris-saline with and without 1mM CaCl2 and 1mM MgCl2 at } pH 6.9,7.2 and 8.0. diluted 1:10-1:100. Next I tried taking a fresh } culture and washing several times in Tris-saline followed by 1:10 } ConA-gold. My incubations have been around 1 hr at room temp. Nary a } gold particle do I see under any of the conditions mentioned. } Apparently, using fluorescently labelled ConA these little buggers } light up like the Hale-Bopp comet, so I am at a lost. Can anybody out } there give me some suggestions. I also have some material fixed in 4% } PF alone, embedded in LRW that I have not done anything yet with. } Thanks in advance, } } Hank Adams } Electron Microscopy Laboratory } New Mexico State University } Las Cruces, NM 88003 } } http://www.nmsu.edu/Research/artsci/public_html/eml/ } } 505-6463600 } } } } \\|// } (o o) } ~~~~~~~~~~~~oOOo~(_)~oOOo~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } { { {This message is made of 100% recycled electrons} } } } } Cheers ;o) :o) %o) } Eric {Mesa Arizona} } http://www.goodnet.com/~earosen (Note the tilde before earosen) }
Etching agents are not usually required for porous acrylics.
If you want to test your fixation, try various ones and then fluorescent labeling--saves a lot of work doing it at the EM level.
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 Microscopy ListServer -- Sponsor: The Microscopy Society of America
Reply to: RE} Sun's Question
What you saw probably were bend extinction contour. (See Hersch et al's book "Electron microscopy of thin crystals", page 203) The change of the pattern may be caused by beam heating.
-------------------------------------- You wrote:
I am a student now using Hitachi-8100 TEM to study submicrometer Cu-Zn alloy particals. The particals are mostly six-fold symmetry plates. When the beam focused on a partical at a high magnification, some dark stripes appear in the partical's bright-field image and change their shapes until a snow-flake-like pattern is formed. The pattern comprised of six center-crossed dark stripes. Select-area diffracton shows a single crystal diffracton pattern with six-fold symmetry. But dark-field image shows that one single diff spot is not coming from the whole partical but from ONE corresponding dark stripe.
My questions are: 1)Do anyone out there ever find similar phenomenon? 2)How can I know the temperature of the particals under the brightening of the electron beam? I think temperature is a reason for the formation of the snow-flake-like pattern.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Subject: Time:4:09 PM OFFICE MEMO Position Available at NCEM Date:3/23/97
Staff Materials Scientist
Materials Sciences Division National Center for Electron Microscopy (NCEM) Job MSD/5001
Essential -- The NCEM is a national user facility with several state-of-the-art electron microscopes and advanced image analysis and specimen preparation facilities, dedicated to the electron-optical microcharacterization of materials. The Center has an opening for a Staff Scientist to lead its research effort in high-resolution imaging of materials. Conduct original research into the development of high-resolution techniques and their application to significant materials problems. Responsible for the operation and development of several high- resolution microscopes, including a new 300kV field emission instrument and the 1MeV ARM. Lead a program to extend instrument resolution toward the 1A information limit.
As an essential member of a national user facility, the candidate will have the opportunity to collaborate with a broad range of investigators from the national and international scientific community. The position also involves close collaboration with other staff members on the development of computer analysis, and control and supervision of postdoctoral and technical staff.
The candidate will be able to contribute substantially to the successful operation and future development of the facility.
QUALIFICATIONS: Essential -- Outstanding track record in development and application of quantitative high-resolution transmission electron microscopy. Extensive practical experience with advanced high-resolution imaging and analysis techniques, such as series reconstruction, holography, and image interpretation. Demonstrated ability to initiate collaborations and carry out high- quality research, using the unique facilities of the NCEM. Marginal -- Ph.D. in physical sciences.
POSTING DATE: March 6, 1997. CLOSING DATE: Open until filled.
MORE INFORMATION:
1. See -- http://www.lbl.gov/LBL-Documents/CJOs/ 2. Go to SCIENTIFIC 3. Go to Job MSD/5001
APPLICATION INSTRUCTIONS (http://www.lbl.gov/LBL-Documents/CJOs/) You may forward your resume or curriculum vitae to: Berkeley Lab Staffing Office One Cyclotron Road, MS 938A Berkeley, CA 94720
Refer to job number MSD/5001 in your cover letter.
E-Mail to: employment-at-LBL.gov
1. Send as plain text (ASCII) in the body of your message. (Attachments or encoded files cannot be read by our automated applicant processing system.) 2. Reference the job number(s) in the SUBJECT of your message. 3. You will not receive an electronic response but will receive a letter of confirmation via the U.S. Postal Service.
Your labelling problem may simply be that you are using resin sections which are usually not permeable to gold labels (ie you only present a very small edge of the bacteria in e.m. whereas I assume the light microscopy that you mention was done on whole cells which present a much larger surface to the fluorescent label. One test might be to try and use the fluorescent label on LR White sections and examine that under the fluorescent light microscope.
Malcolm Haswell University of Sunderland UK ----------
Dear Sun, As someone else stated, the dark lines you saw in bright field are bend contours. You saw three crossed bend contours, each which corresponds to a particular diffraction condition. Thus where all three bend contours cross, all three diffraction conditions are operating simultaneously and you are looking down the zone axis of the crystal. When you work in dark field you will be imaging with only one of the diffraction spots so you will only see one bend contour (which will now be bright). As for why the pattern changes on observation, this is quite common and you will find that many things deform or tilt slightly under the heat of the beam. I don't know about your system but in some systems, the heating can induce crystallisation which would obviously change the contrast pattern. Lastly, I can't think of a simple way of measuring the temperature of your particles.
Yours sincerely
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
If you have full World Wide Web access you could try using a search engine such as Lycos or Yahoo (eg microscope, microscopy, amateur, hobby). It should turn up some useful web pages which could lead to further links.
One really excellent on-line organization & magazine is: Microscopy-uk http://www.microscopy-uk.org.uk/#top which has the online magazine 'Micscape'. It has had excellent articles on everything from hand lenses to electron microscopes and makes few assumptions that it's readership are pro microscopists. There are also lots of links.
Other places to try would be the home pages of the Microscopy Society of America (US) and Royal Microscopy Society (UK) and again look for useful links.
I hope this helps
Malcolm Haswell University of Sunderland UK
----------
Can any one tell me of a Microscope news group taylored toward the newbie into this hobby? Also, are there any good magazines on the market specializing in MIcroscopes and the Microscope hobby?
I'd like to thank Marks, Dorai, Garber, Yi Huang and Ian MacLaren for their suggestions to my questions.
Bend contours and multiply twins are possible reasons for the formation of the pattern described in my questions. I will find more details on these matters.
A commercial source for Aclar is ProPlastics, Inc, P.O. Box 679, Linden, NJ 07036 201-925-5555. They have a minimum order. Several years ago we bought two pounds of 5mil for $75.00. It will be a lifetime supply.
At 03:55 PM 3/21/97 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
Hi Gary, Aclar is great! Read the paper in Journal of Electron Microscopy Technique 10:77-85 (1988), by Kingsley and Cole. I have used it in epon embedding protocols. There is nothing like it. You can get it from Electron Microscopy Sciences. Call them since it may not be listed in their catalogue. Ask to speak to Stacie Kirsch.
Aclar is a sturdy thin film, transparent as glass, no detectable autofluorescence, chemically inert, sections beautifully, and so on and so on.
Have fun.
Sally
On Fri, 21 Mar 1997, Gary Radice wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Several of you have recommending using Aclar film in your UV polymerization } protocols. What is Aclar film, and where do you purchase it? } } Gary Radice, Associate Professor gradice-at-richmond.edu } Department of Biology 804-289-8107 (voice) } University of Richmond VA 23173 804-289-8233 (FAX) } } }
(I am posting this for a colleague in France who is not on this list. Contact him, not me, please.) ==============================
My laboratory has bought recently a SX-100, so I should be happy to find somebody who might be interested in purchasing our Camebax Electron Microprobe, which is in good condition.
Here is a short description : Camebax with 3 WDS and 1 EDS for fully compatible coupled analysis WDS-EDS (Quantitative analysis on fixed points or lines, X-ray mapping) WDS : all usual crystals, plus PC1 and PC2 for light elements EDS : Tracor model, with Be window Equiped with Back-Scattered Electron detector, and anticontamination system. Software : all the software provided by Tracor (Noran) for TN-5500 : - to control spectrometers, sample holder, and faraday cup - to perform quantitative analysis (ZAF and PRZ) - to make standardless analysis (EDS) - Image analysis (by digitalisation)
The column is from 1976, but everything else is from 1988 (including spectrometers)
Price = $30 000 USD. (shipping to be paid by purchaser)
For more information, contact:
Michel LAHAYE ICMCB, Universiti Bordeaux I, Pessac (FRANCE) Tel 33 5 56 84 62 93 Fax 33 5 56 84 27 61 e-mail : lahaye-at-icmcb.u-bordeaux.fr ==================================================================
Jussi: We could only find Swift point counters when we were in the market for a new counter for mineralogical applications. They were still in business 5 years ago when we bought our newer electronic model. There may be other companies selling similar products. We had an ancient one (} 35 years)that with maintenance the stage worked, but the old electric pushbuttons were in bad shape. It is dependable and a very adaptable system. If you get all the gear sets, you can count at any optimum step size typical of optical microscope use. We used it primarily for opague ore minerals in polished briquettes. The U.S. office is in San Jose CA. Fax 408-292-7967. I'm sure there is a closer one to you in Europe, but I don't have any info on it. You might try looking on the WWW for a webpage.
I am presently unemployed and have no financial interest in Swift or anything at present, but would be glad to have some! ================================================ Michael L. Boucher Sr. mboucher-at-isd.net 13345 Foliage Avenue Apple Valley, MN 55124-5603 Ph 612-432-8836 ================================================
On Thu, 20 Mar 1997, Ginger Baker wrote: } } Hello all, } } I have a PhD student from Brazil who is studying Catolaccus } grandis. He places a female on top of a beeswax cell which contains } boll weevil larva. The female deposits an egg on the larva and 2 } weeks later, it matures and burrows its way out of the beeswax } creating a hole. He would like to study and measure these holes. } } However, from past experience I know that beeswax melts when placed } in my Au/Pd sputter coater. Is there any way to coat the beeswax and } examine them with an SEM? Or should he try another route? } } Thank you in advance,
} Email: lizard-at-okway.okstate.edu } Hi Ginger, I guess the coater you used is a diode type of sputter coater. Heat is generated during coating. In order to solve your problem, you should to use a kind of "cooled sputter coater"-- planar magnetron sputter (PMS) coater. A permanent magnet is posstioned at the center of the cathode to deflect the electrons away from the specimen.
On Thu, 20 Mar 1997, Ginger Baker wrote: } } Hello all, } } I have a PhD student from Brazil who is studying Catolaccus } grandis. He places a female on top of a beeswax cell which contains } boll weevil larva. The female deposits an egg on the larva and 2 } weeks later, it matures and burrows its way out of the beeswax } creating a hole. He would like to study and measure these holes. } } However, from past experience I know that beeswax melts when placed } in my Au/Pd sputter coater. Is there any way to coat the beeswax and } examine them with an SEM? Or should he try another route? } } Thank you in advance,
} Email: lizard-at-okway.okstate.edu } Hi Ginger, I guess the coater you used is a diode type of sputter coater. Heat is generated during coating. In order to solve your problem, you should to use a kind of "cooled sputter coater"-- planar magnetron sputter (PMS) coater. A permanent magnet is positioned at the center of the cathode to deflect the electrons away from the specimen.
Ya Chen
Ya Chen
========================================================================= \ / Integrated Microscopy Resource (IMR)-- \ / __ an NIH Biomedical Research Resource TEL : 608-263-8481 \/ / / University of Wisconsin-Madison FAX : 608-265-4076 / / / 1675 Observatory Drive #159 Email1:ychen14-at-facstaff.wisc.edu / /__/_ Madison, WI 53706 Email2:chen-at-calshp.cals.wisc.edu ========================================================================= IMR WWW Home Page: http://www.bocklabs.wisc.edu/imr.html
I am looking for a source of adapters to convert the old short barrel Leitz objectives to the newer long barrel type (170mm tube length).
I also need a set of PLAN APO's for a Zeiss Photomicroscope in 160mm tube length. Need 4X, 10X, 20X, 40X and 100X O.I. or anything close in Plan Apo's.
Also need the PHACO 2 and PHACO 3 inserts for a Leitz Laborlux-11 (type 55 condenser).
Any leads would be most appreciated.
Cordially, Gil Groehn (313) 884-1139
-- ============================================================= ULTRAMED, INC. Research Div. ad408-at-detroit.freenet.org Grosse Pointe Farms, MICH 48236 USA PHONE: 313-884-1139 ===============================================================
Reprosil is good, and so are any of the other polyvinyl siloxane impression materials, available from any dental supplier. The resin can be Epon, or 302-1 from Epo-Tek or anything from the EM world.
Lesley Weston
On Fri, 21 Mar 1997, Ronald LHerault wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } An alternative may be replication using Reprosil, a Polyvinyl Siloxane } from Caulk. It makes a negative which can be transfered to a positive by } using Spurr low viscosity embedding medium. Detail is there albeit a } little softened. } } Ron } }
I was looking at your page and hoped you might help me. I am looking for a Windows based SEM capture board. We do quite a bit of optical microscope image capture, but nothing yet on SEMs.
I have heard there are a couple out there, but haven't found them yet!
Good afternoon, I was wondering if those of you who successfully make your own holey grids for stigmation would care to share the method. I've been using glcerol and 0.25% Formvar (old bottle) and while I can produce small, nice looking "holes", on close inspection they're film. There are some true holes, but few and far between. Any suggestions? Many thanks in advance.
Dwight Beebe E-mail: beebed-at-ere.umontreal.ca Institut de recherche en biologie vegetale Voice: 514-872-4563 Universite de Montreal FAX: 514-872-9406 4101, rue Sherbrooke est Montreal, Quebec H1X 2B2 Canada
We historically have made our own holey films, because we could not buy good holey support films from any manufacturer. Without question, it is an art, and most of the time a major pain in a posterior region. I would be happy to send you the detailed instructions we have found to be most reproducible.
Recently, however, we purchased some holey films from SPI, which have been uniformly *gorgeous*. They are thin, clean, a large fraction of holes etc. Probably equivalent to the best I have ever made. I don't recall the price, but my impression is that I can't make them as cheaply, so why bother.
Highly recommended.
Larry
PS I have no particular connection to SPI.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Dwight, I know of two methods of preparing holey films. The first is to shake up a mixture of liquid soap and water and Collodion until it froths, then drop a drop or two on the surface of distilled water. Lay the grids on top of the Collodion layer, pick up with a filter paper, dry and carbon coat. Put the grids in a Jaffe washer with cloroform for 48 hours to remove the Collodion. The other method, if you have Nucleopore type filters, is to carbon coat a strip of Nucleopore filter film, place a square of the coated filter on the grid sitting on a nickel mesh on the surface of the Jaffe washer full of cloroform, wait 48 hours to dissolve the Nucleopore, dry grids. The holes in the Nucleopore will be now be holes in the carbon coat. I must admit I have not tried the first method myself, but someone in my lab did, years ago. You wrote: } I was wondering if those of you who successfully make your own } holey grids for stigmation would care to share the method. I've been using } glcerol and 0.25% Formvar (old bottle) and while I can produce small, nice } looking "holes", on close inspection they're film. There are some true } holes, but few and far between. Any suggestions? Many thanks in advance. } } } Dwight Beebe E-mail: beebed-at-ere.umontreal.ca } Institut de recherche en biologie vegetale Voice: 514-872-4563 } Universite de Montreal FAX: 514-872-9406 } 4101, rue Sherbrooke est } Montreal, Quebec H1X 2B2 } Canada Good luck, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
I am looking for any information on the fluorescent dye: Commercial name; 'Geranine G', C.I. name; Direct Red 48, C.I. number; 14930. I know that it is a monoazo dye with good affinity for certain proteins, so my question is does anyone have any further information on Geranine G, such as Excitation and Emission wavelengths, or even manufacturers or distributors that carry this dye. Failing this would anyone know of a substitute of similar properties.
{fontfamily} {param} Times_New_Roman {/param} {smaller} One Post-Doctoral Position and one Graduate Assistantships Available at the {/smaller} {/fontfamily} {smaller} {fontfamily} {param} Times {/param}
{/fontfamily} {fontfamily} {param} Times_New_Roman {/param} University of Michigan. {/fontfamily} {fontfamily} {param} Times {/param}
{/fontfamily} {fontfamily} {param} Times_New_Roman {/param} We are in search of a Post-Doctoral Scholar to carry out research {/fontfamily} {fontfamily} {param} Times {/param}
{/fontfamily} {fontfamily} {param} Times_New_Roman {/param} in the areas of processing and TEM characterization of functional ceramics, {/fontfamily} {fontfamily} {param} Times {/param}
{/fontfamily} {fontfamily} {param} Times_New_Roman {/param} including thin films, thick films and bulk materials. Candidates should have {/fontfamily} {fontfamily} {param} Times {/param}
{/fontfamily} {fontfamily} {param} Times_New_Roman {/param} a Ph.D. in a physics- or materials-related field, with outstanding academic and {/fontfamily} {fontfamily} {param} Times {/param}
{/fontfamily} {fontfamily} {param} Times_New_Roman {/param} publication records. Hands-on experience in the areas of thin thin processing and {/fontfamily} {fontfamily} {param} Times {/param}
{/fontfamily} {fontfamily} {param} Times_New_Roman {/param} TEM (HRTEM and/or AEM) characterization of crystal defects and interfaces in {/fontfamily} {fontfamily} {param} Times {/param}
{/fontfamily} {fontfamily} {param} Times_New_Roman {/param} the material is highly desirable. This position is available immediately for one year, {/fontfamily} {fontfamily} {param} Times {/param}
{/fontfamily} {fontfamily} {param} Times_New_Roman {/param} with the possibility of being renewed for an additional two years. The salary level {/fontfamily} {fontfamily} {param} Times {/param}
{/fontfamily} {fontfamily} {param} Times_New_Roman {/param} will be commensurate with qualifications and experience. {/fontfamily} {fontfamily} {param} Times {/param}
{/fontfamily} {fontfamily} {param} Times_New_Roman {/param} We are also in search of one Graduate Research Associate to pursue Ph.D. in {/fontfamily} {fontfamily} {param} Times {/param}
{/fontfamily} {fontfamily} {param} Times_New_Roman {/param} the general field of functional ceramics, starting Summer/Fall 1997. The {/fontfamily} {fontfamily} {param} Times {/param}
{/fontfamily} {fontfamily} {param} Times_New_Roman {/param} candidates should have a M.S. in a materials-related field, with an excellent {/fontfamily} {fontfamily} {param} Times {/param}
{/fontfamily} {fontfamily} {param} Times_New_Roman {/param} academic=20 record. The assistantship carries with an attractive stipend (~16,3k), {/fontfamily} {fontfamily} {param} Times {/param}
{/fontfamily} {fontfamily} {param} Times_New_Roman {/param} a full-payment of tuition, and benefits. {/fontfamily} {fontfamily} {param} Times {/param}
{/fontfamily} {fontfamily} {param} Times_New_Roman {/param} =20 Interested candidates should contact Prof. Pan, preferably by Email: {/fontfamily} {fontfamily} {param} Times {/param}
{/fontfamily} {fontfamily} {param} Times_New_Roman {/param} Prof. Xiaoqing Pan {/fontfamily} {fontfamily} {param} Times {/param}
{/fontfamily} {fontfamily} {param} Times_New_Roman {/param} Department of Materials Science and Engineering {/fontfamily} {fontfamily} {param} Times {/param}
{/fontfamily} {fontfamily} {param} Times_New_Roman {/param} University of Michigan {/fontfamily} {fontfamily} {param} Times {/param}
{/fontfamily} {fontfamily} {param} Times_New_Roman {/param} Ann Arbor, MI-48109-2300 {/fontfamily} {fontfamily} {param} Times {/param}
Ron Herault wrote: ----------------------------------------- An alternative may be replication using Reprosil, a Polyvinyl Siloxane from Caulk. It makes a negative which can be transfered to a positive by using Spurr low viscosity embedding medium. Detail is there albeit a little softened. ----------------------------------------- Another alternative, possibly an even better choice, would be our own "Wet Surface Replica Kit", it too is silicone based, but it has been more optimized for SEM applications. It can be cured into a "negative" much more quickly, and when converted, the "positive" replicating system is much easier to use than the "Spurr" approach and on a "per replica basis" is a lot lower in cost. A demo set of micrographs made from such positive replicas, on human skin, can be seen in our electronic catalog at the website mentioned below.
Disclaimer: SPI Supplies manufactures the above mentioned "Wet Replica Kit" and would have a vested interest in seeing more persons using it.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
In message {199703241633_MC2-1336-770C-at-compuserve.com} , "Seth J. Grotelueschen" {sethg-at-CompuServe.COM} writes
Dear Seth
Is is not obvious which country you are from but if you are interested in someone in the UK try Deben Research who will be able to help you. If you want the full address and contact no let me know.
They also support our imaging software.
Best regards
Dominique } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
---------------------------------------------------------------------------- |From: Dominique Miller | | |Marketing Executive | | |Foster Findlay Associates | Phone: National (0191) 201 2180 | |Newcastle Technopole | International +44 191 201 2180 | |Kings Manor | Fax: National (0191) 201 2190 | |Newcastle upon Tyne | International +44 191 201 2190 | |NE1 6PA |E-Mail: Dominique-at-ffaltd.demon.co.uk | |UK | WWW: http://www.demon.co.uk/ffaltd/ | ----------------------------------------------------------------------------
We are studying some ultrastructural features of a new type of epidermal trichomes on plants. Trying to define and quantify ulstrastructural changes during the cell development (from meristematic stage to fully differentiated trichome) we have to deal with the following questions. How to start analysing TEM micrographs in order to reach general conclusions on variations of organelle numbers ? Are there easy methods to start quantitative morphological analysis ? Do we have to work with image analysing softwares or not ? How do you deal with this kind of problem ? Do you know about books or papers on similar studies that can be used as references?
Any solutions are welcome... Thanks !
************************************ Pascal VEYS Laboratory of Plant Biology Catholic University of Louvain Place Croix du Sud 5 (bte 14) B 1348 Louvain-la-Neuve Belgium Phone : 0032 10473004 Fax : 0032 10473471 Email : Veys-at-bota.ucl.ac.be ************************************
ULTRAMICROTOMY ANY CRYO METHODS FOR MATERIALS SCIENCE
PLACE: UNIVERSITY OF PENNSYLVANIA LABORATORY FOR THE RESEARCH ON THE STRUCTURE OF MATTER PHILADELPHIA, PA 19104 DATE: JUNE 11-13,1997 SPONSORS: LEICA INC., DIATOME U.S., ELECTRON MICROSCOPY SCIENCES
COURSE SPEAKERS AND INSTRUCTORS: Dr. Tom Malis, Phil Swab, Helmut Gnagi
OVERVIEW:
This workshop will cover the use of Room Temperature and CryoUltramicrotomy techniques as it applies to Material Science. Included in the course will be all of the preparation leading up to sectioning, such as, embedding, trimming and staining on different types of industrial specimens(polymers, metals, plastics, fibers, rubbers, powders, foils, etc.). From all of the topics to be covered and in depth review of all of the different techniques will take place and there will be an emphasis on "Hands-On" lab time. Participants are encouraged to bring their own samples as well. Our goal, upon completion of the course, is that each of the participants will be able to return to there own lab with a better understanding of the theory and principles behind ultramicrotomy, and the ability to successfully perform all of the techniques which we shall cover.
COURSE COST: $1,500.00 The price includes: Hotel accommodations(3 nights), lunch daily, continental breakfast daily, 1 group dinner, all course supplies and full lab time.
FIRST COURSE DEADLINE: April 18, 1997
Parties that are interested may either respond by E-Mail to SGKCCK-at-AOL.COM or Leica's seminar voice mail to 1-800-248-0665 EXT:5010 Contacts are Stacie Kirsch or Ann Korsen. To confirm your space in the course below is a form that may be filled out and either E-Mailed to SGKCCK-at-aol.com or faxed to 215-646-8931 ATT: Stacie Kirsch. All checks should be mailed to Diatome U.S. P.O. Box 125, Fort Washington, Pa 19034 and made out to Leica, Inc.
WORKSHOP APPLICATION FORM: Company Name:________________________ Participant's Nmae:______________________ Full Address:____________________________ City, State, Zip:__________________________ Tel #:___________________________________ Fax #:___________________________________ E-Mail:__________________________________ Best Time To Reach:______________________ Area of Interest: Cryo or Room Temperature Type of Material Working With:______________ Will you be bringing your own sample?___________________What:________
Paying by PO#:________________________(Please mail to P.O. Box) Check#:_______________________________
If paying by Purchase order number please be sure to include your full billing address.
The normal replicas of ruled gratings allow SEM magnification calibration up to about 50 000x mag. At magnifications higher than this it is not so easy to find good accurate SEM calibration specimens.
What SEM calibration specimens are available for use at 50 000 to 1 000 000 x mag in an In-lens FEGSEM?
Regards,
Dr Jan Coetzee Head: Unit for Electron Microscopy Tel:+27-12-420-2075 University of Pretoria Fax:+27-12-342-1738 Pretoria 0002 Internet:janc-at-ccnet.up.ac.za South Africa http://www.up.ac.za/academic/electron/emunit1.htm
Hello, A recent upgrade in EDS instrumentation and detectors has given us the ability to detect low elements (C, N, O). I have been detecting a peak that corresponds to nitrogen in all the titanium spectra that I have acquired. This has occurred using pure titanium (T18) or alloyed titanium (beta ti). The nitrogen peak ranges from 5-10 weight percent of the spectra. I know that certain elements can implant in Ti, including C and N, but these samples have not undergone the heating conditions that would favor ion implantation. The N peaks do not increase under long e-beam bombardment. The spectra are collected under routine conditions (15KeV, recommended WD, 25% deadtime, 100 sec, standardless analysis but the calibration seems correct). I would appreciate it if anyone could shed some light on this problem.
Wallace Ambrose Dental Research Center Univ. of North Carolina Chapel Hill, NC
The 9th ed of Conn's Biological Stains offers little on Geranine G, only a few references. Try the Biological Stain Commission, Dept. of Pathology, Univ. of Rochester Medical Center, Rochester, NY (716)-275-2751.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (908)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
Ambrose, Wallace wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello, } A recent upgrade in EDS instrumentation and detectors has given us the } ability to detect low elements (C, N, O). I have been detecting a peak } that corresponds to nitrogen in all the titanium spectra that I have } acquired. This has occurred using pure titanium (T18) or alloyed titanium } (beta ti). The nitrogen peak ranges from 5-10 weight percent of the } spectra. I know that certain elements can implant in Ti, including C and } N, but these samples have not undergone the heating conditions that would } favor ion implantation. The N peaks do not increase under long e-beam } bombardment. The spectra are collected under routine conditions (15KeV, } recommended WD, 25% deadtime, 100 sec, standardless analysis but the } calibration seems correct). I would appreciate it if anyone could shed } some light on this problem. } } Wallace Ambrose } Dental Research Center } Univ. of North Carolina } Chapel Hill, NC
Just a quick thought: are you sure that it is a nitrogen peak and not Ti L, or Ti L Escape peak? --
Naresh Shah Associate Research Professor, University of Kentucky Department of Chemical and Materials Engineering (CME) Consortium for Fossil Fuel Liquefaction Science (CFFLS) 533 South Limestone Street, Room 111 Lexington, KY 40508-4005 Phone: (606) 257-5119; FAX: (606) 257-7215; e-mail: naresh-at-pop.uky.edu
} A recent upgrade in EDS instrumentation and detectors has given us the } ability to detect low elements (C, N, O). I have been detecting a peak } that corresponds to nitrogen in all the titanium spectra that I have } acquired.
This is the Ti L-l line (the transition of the M-I to the L-III subshell), a throroughly nasty interference, even in WDS. Contact me if you have problems with it.
Alfred
----------------------------------------- Alfred Kracher akracher-at-iastate.edu http://www.public.iastate.edu/~akracher -----------------------------------------
If your SEM screen accepts a Polaroid camera, Polaroid makes a "MicroCam" and "PhotoPad" color scanner. Together they retail for about $1000. I have only seen the ad, and I have no first hand information (Polaroid: 1-800-662-8337). I have no connection to or investment in Polaroid.
There are other manufacturers of frame-grabbers and videoboards for SEMs, but alas, I discarded the information.
Scott Schwinge Friday Harbor Labs University of Washington
} Hi, } } I was looking at your page and hoped you might help me. } I am looking for a Windows based SEM capture board. We } do quite a bit of optical microscope image capture, but } nothing yet on SEMs. } } I have heard there are a couple out there, but haven't } found them yet! } } Thanks } Seth Grotelueschen
Can anyone suggest names of service engineers with experience on vintage Cambridge electron microscopes? I have a 1971 Cambridge Stereoscan S4 SEM in need of service. I'd like to locate an experienced technician in the metro Atlanta area or the southeastern US, but I would appreciate all references and information. Thanks,
Michael Knotts ------------------------------------------------------------- Michael E. Knotts, Ph.D. E-mail: ph281mk-at-prism.gatech.edu Contributing Editor {The Light Touch} OPTICS & PHOTONICS NEWS Georgia Tech / School of Physics / Atlanta, GA 30332-0430 Tel: (404) 894-3422 FAX: (404) 894-9958
Although titanium readily attracts nitrogen and oxygen, I think your problem is related to peak overlap.
A quick look at x-ray tables reveals:
El/line keV
N Ka1&2 0.392
Ti Ll 0.395
Ti La1&2 0.452
O Ka1&2 0.523
The N Ka and Ti Ll peaks cannot be resolved by typical EDS spectrometers, since you would need a resolution of ~2 ev. To resovle the N Ka and Ti La peaks you would need a detector resolution of ~50 ev.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
electron-sample interaction programs Hello, I am in need of a Monte Carlo based program (PC or Mac) that models the interaction of an electron beam with a user defined sample. We have a 200 keV TEM and an SEM capable of imaging at low voltage; hence, the program should handle electron transparent samples up to bulk materials and a ~1 to 200 keV electron beam of variable probe size. I am not only interested in the electron interaction volume but also in the volume from which many other signals can be collected, e.g., backscattered electrons, x-rays, secondary electrons, Auger electrons, etc.
I have seen electron flight simulator for sale ($500), but have little info on what it provides. What about quality shareware?
Thank you
Brad Storey Materials Scientist Argonne National Lab 208-533-7685
} A recent upgrade in EDS instrumentation and detectors has given us the } ability to detect low elements (C, N, O). I have been detecting a peak } that corresponds to nitrogen in all the titanium spectra that I have } acquired. This has occurred using pure titanium (T18) or alloyed titanium } (beta ti). The nitrogen peak ranges from 5-10 weight percent of the
This is not a nitrogen peak. You are seeing the Ti L-series lines, which exhibit a strong overlap with the Nitrogen K-series lines.
This overlap is bad enough that it cannot really be easily resolved even using a wavelength-dispersive spectrometer. It is one of the classic overlaps that makes analysis of Ti alloys for nitrogen problematic.
Nitrogen detection is hampered by strong absorption of Ni Ka by any carbon that is in the x-ray path, be it in the sample, the carbon coat on the specimen, oil on the EDS detector window, window material (like diamond, for example), etc.
These factors make nitrogen measurement a challenge, since one does not expect to find it in any real concentration unless it is a distinct phase (like TiN).
Paul
+----------------------------------------------------+ | Paul K. Carpenter paulc-at-gps.caltech.edu | | Division Analytical Facility | | Geological and Planetary Sciences MC 100-23 | | California Institute of Technology | | Pasadena, CA 91125 | | 818-395-6126 (X-ray Lab) 818-568-0935 (Dept. FAX) | +----------------------------------------------------+
I use a method by Kuga and Brown from Philips Electron Optics Bulletin v126, p.19 (1989) to make lacy formvar grids. First a note of caution. I use dichloroethane as my solvent for the formvar resin. After having much difficulty in preparing my films, I was told that when exposed to light, dichloroethane slowly forms HCl in the solution. The HCl destroys the integrity of the formvar polymer. Solution: store the formvar solution in a brown bottle in a dark cabinet.
Method:
- I use 0.25% formvar solution in dichloroethane and freshly cleaved mica sheets as a substrate. The formvar lifts off the mica much easier than from glass microscope slides.
- Dip the mica into the formvar solution, wick off excess on a paper towel, then breathe heavily on the mica for about 5 seconds while it is still wet. The moisture in your breath condenses in the solution. Caution: Don't inhale!
- When the mica has dried, score the edges and float off on water.
- Place 200 mesh TEM grids on the floating film. The area with the best holes will appear milky.
- I then take saran wrap, stretch it tightly across the mouth of a small (~100ml beaker), and press it down at a slight angle onto the floating formvar film. The film will stick to saran wrap and you can easily pick it up off the surface of the water.
- After the film has mostly dried, pick up the grids from the saran wrap "drumhead" and place them on filter paper.
- The film will have many "pseudoholes" that have a thin residual film across them.
- Following the method of Kuga and Brown, by heating the film to about the T(g) temperature you can break these holes open. The time and temperature are critical. I place the bare filter paper in a small lab oven (not in a petri dish; it has too much thermal mass) at 110C for 12 minutes.
- I then coat both sides with carbon to stabilize the formvar lace.
I can easily make 50-100 grids in an hour (exclusive of the carbon coating). These grids make wonderful supports for looking at fine particulate dispersions.
Cheers, Henk
} } Good afternoon, } I was wondering if those of you who successfully make your own } holey grids for stigmation would care to share the method. I've been using } glcerol and 0.25% Formvar (old bottle) and while I can produce small, nice } looking "holes", on close inspection they're film. There are some true } holes, but few and far between. Any suggestions? Many thanks in advance. } } } Dwight Beebe E-mail: beebed-at-ere.umontreal.ca } Institut de recherche en biologie vegetale Voice: 514-872-4563 } Universite de Montreal FAX: 514-872-9406 } 4101, rue Sherbrooke est } Montreal, Quebec H1X 2B2 } Canada
Henk Colijn colijn.1-at-osu.edu OSU Campus Electron Optics Facility ------------------------------------------------------------------- Prosperity is the blessing of the Old Testament; Adversity is the blessing of the New. Francis Bacon, "Of Adversity."
Does anyone know of a source for teflon o-rings with a thickness of {100 uM?
Thanks, Sandy Simon
Sanford M. Simon Laboratory of Cellular Biophysics Box 304 Rockefeller University 1230 York Avenue New York, N.Y. 10021 212-327-8130 (voice) 212-327-8022 (fax) simon-at-rockvax.rockefeller.edu (e-mail)
Does anyone have a current phone number for Energy Beam Sciences or whichever company has taken over Polaron? I have a question about our Polaron film thickness monitor. Thanks for your help.
} Dear All, } } The normal replicas of ruled gratings allow SEM magnification } calibration up to about 50 000x mag. At magnifications higher than } this it is not so easy to find good accurate SEM calibration } specimens. } } What SEM calibration specimens are available for use at } 50 000 to 1 000 000 x mag in an In-lens FEGSEM? } } Regards, } } } } Dr Jan Coetzee
In biological TEM, colloidal gold in a variety of immunological labeling techniques. This gold has fairly well defined size distributions - the smallest available has a mean size of 1nm. If you check the EM supplies catalogues (Agar, SPI, Ted Pella, etc), you should find it - personally, I've never tried it, but I don't see any reason why it couldn't be used (although you might need to remove some antibodies).
The other approach is to use the same techniques that is used in TEM, where ther is a bit of a gap between gratings and lattice resolution specimens. Basically, you work up step by step, going from a lower mag, where the grating gives you a calibration, identify a number of obvious features, then go up in mag and relocate the same features - they got to be the same distance apart. A bit tedious and not statistically very accurate, +/- 5% at best, but its better than nothing.
Brad Storey wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } electron-sample interaction programs } Hello, } I am in need of a Monte Carlo based program (PC or Mac) that models the } interaction of an electron beam with a user defined sample. We have a 200 keV } TEM and an SEM capable of imaging at low voltage; hence, the program should } handle electron transparent samples up to bulk materials and a ~1 to 200 keV } electron beam of variable probe size. I am not only interested in the electron } interaction volume but also in the volume from which many other signals can be } collected, e.g., backscattered electrons, x-rays, secondary electrons, Auger } electrons, etc. } } I have seen electron flight simulator for sale ($500), but have little info on } what it provides. What about quality shareware? } } Thank you } } Brad Storey } Materials Scientist } Argonne National Lab } 208-533-7685
On my laboratory web site are informations about almost all public domain and commercial Monte Carlo programs for PC and Mac's.
} Hello, } A recent upgrade in EDS instrumentation and detectors has given us the } ability to detect low elements (C, N, O). I have been detecting a peak } that corresponds to nitrogen in all the titanium spectra that I have
snips
} calibration seems correct). I would appreciate it if anyone could shed } some light on this problem. } } Wallace Ambrose } Dental Research Center } Univ. of North Carolina } Chapel Hill, NC
Can anyone refer me to a basic reference on electron channeling = contrast, or failing that, give me a few hints on optimal conditions for = observing this type of contrast ?=20
TIA=20
Glenn
Glenn Poirier Phone (514) 398 6774 Electron Microprobe Laboratory Fax (514) 398 4680 Earth and planetary Science=20 Mcgill University
THERE ARE THREE SIDES TO EVERY STORY: yOURS, MINE AND THE TRUTH
Talking of peak overlaps, is anyone out there capable of reliable accurate analysis of V in the presence of large amounts of Ti, using:
a WDS
b EDS?
I ask because I concluded recently that I couldn't, with my EDS-only system, analyse for V in titanomagnetites, and that it probably wasn't possible even with WDS. If I ask my system to look for V in my standard Rutile (TiO2), it always finds a few tenths of a %, even though my software (LINK ZAF4/FLS) strips out Ti Kb along with the Ka. I have no way of telling whether this V is genuine or not, but I suspect that it isn't, as my supposedly-pure Ti metal standard also gets credited with a small amount of V, which I doubt. I was recently given another rutile standard, and, blow me down, it is reputed to have 0.4% V by WDS analysis. Anyone got any feeling for whether this is likely to be true?
cheers
Ritchie
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
Wallace: This is most likely the Ti Ll line overlapping N Ka. Beware of overlaps in the light element energy region!
Jim *.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.*.* James J. McGee (jmcgee-at-sc.edu) Dept. of Geological Sciences University of South Carolina (803) 777-6300 (Office) Columbia, SC 29208 (803) 777-6610 (Fax)
----------
Hello, A recent upgrade in EDS instrumentation and detectors has given us the ability to detect low elements (C, N, O). I have been detecting a peak that corresponds to nitrogen in all the titanium spectra that I have acquired. This has occurred using pure titanium (T18) or alloyed titanium (beta ti). The nitrogen peak ranges from 5-10 weight percent of the spectra. I know that certain elements can implant in Ti, including C and N, but these samples have not undergone the heating conditions that would favor ion implantation. The N peaks do not increase under long e-beam bombardment. The spectra are collected under routine conditions (15KeV, recommended WD, 25% deadtime, 100 sec, standardless analysis but the calibration seems correct). I would appreciate it if anyone could shed some light on this problem.
Wallace Ambrose Dental Research Center Univ. of North Carolina Chapel Hill, NC
The question of how to make holey carbon films came up on the list server two years ago. At that time John Gabrovsek (gabrovj-at-ccsmtp.ccf.org) gave the following reference: Baumeister & Seredynsky, Micron 1976, Vol 7, p. 49, and Jane Fagerland (fagerland.jane-at-igate.abbott.com)gave this one: Elsner, Proceedings, 29th EMSA Meeting, p. 460. Both methods were said to work satisfactorily. You might try contacting these people for more details.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
Dear pascal veys: Here are some basic references that may help you get started: Weibel, E.R. 1979. Stereological methods, vol 1. Practical Methods for biological morphometry. Academic Press, New York.
Weibel, E.R., "Stereological Techniques for Electron Microscopic Morphometry". In Principles and Techniques of Electron Microscopy, Vol. 3, M.A. Hayat, ed. 1973. pp 239-296.
Loud, A.V., 1968. A quantitative stereological description of the ultrastructure of normal rat liver parenchymal cells. Journal of Cell Biology, Vol 37:27-46.
Don Gantz Boston Univ Med School gantz-at-med-biophd,bu.edu
Hi Seth, JEOL in Scandinavia market a system called Semafore, their Web page may be worth a look as it has sample software to download. email me if you would like more information.
all the best Pete Lander (JEOL UK)
} -----------------------. } } } } Hi, } } } } I was looking at your page and hoped you might help me. I am looking for a } } Windows based SEM capture board. We do quite a bit of optical microscope } } image capture, but nothing yet on SEMs. } } } } I have heard there are a couple out there, but haven't found them yet! } } } } Thanks } } Seth Grotelueschen }
Return messages: Home Work email pal-at-fenland.demon.co.uk pal-at-jeolsys.demon.co.uk phone 01354 661413 01707 377117 fax phone first 01707 373255
The MicroCam slides onto the eyepiece of a optical microscope. It doesn't work with a SEM camera port. John D. Warren Area Sales Manager Digital Products Polaroid Corporation
4525 Leonard Parkway Richmond, Virginia 23221-1809
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
If your SEM screen accepts a Polaroid camera, Polaroid makes a "MicroCam" and "PhotoPad" color scanner. Together they retail for about $1000. I have only seen the ad, and I have no first hand information (Polaroid: 1-800-662-8337). I have no connection to or investment in Polaroid.
There are other manufacturers of frame-grabbers and videoboards for SEMs, but alas, I discarded the information.
Scott Schwinge Friday Harbor Labs University of Washington
} Hi, } } I was looking at your page and hoped you might help me. } I am looking for a Windows based SEM capture board. We } do quite a bit of optical microscope image capture, but } nothing yet on SEMs. } } I have heard there are a couple out there, but haven't } found them yet! } } Thanks } Seth Grotelueschen
Don't feel alone. Last year, I tried to do a quantitative analysis of 6/4 titanium/vanadium by EDS. The responses you have received so far explain the problem - peak overlaps. I had the additional factor of a V L-alpha emission at 0.51 keV. I opted for emission spectroscopy at an outside lab.
Regards,
-Bob **************************** Bob Citron Chiron Vision Claremont, CA Bob_Citron-at-cc.chiron.com ****************************
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello, A recent upgrade in EDS instrumentation and detectors has given us the ability to detect low elements (C, N, O). I have been detecting a peak that corresponds to nitrogen in all the titanium spectra that I have acquired. This has occurred using pure titanium (T18) or alloyed titanium (beta ti). The nitrogen peak ranges from 5-10 weight percent of the spectra. I know that certain elements can implant in Ti, including C and N, but these samples have not undergone the heating conditions that would favor ion implantation. The N peaks do not increase under long e-beam bombardment. The spectra are collected under routine conditions (15KeV, recommended WD, 25% deadtime, 100 sec, standardless analysis but the calibration seems correct). I would appreciate it if anyone could shed some light on this problem.
Wallace Ambrose Dental Research Center Univ. of North Carolina Chapel Hill, NC
Does anyone have a current phone number for Energy Beam Sciences or whichever company has taken over Polaron? I have a question about our Polaron film thickness monitor. Thanks for your help.
Hi there: My name is Miguel Sanchez my problem is the following As part of my business I receive micron rated textiles, that is nylon mesh that has a determined distance between threads. Sometimes is very difficult to determine that the textile is inside tolerances or not. I use an optical microscope to do measure them my question is: What do I need to measure the textiles with the use of a camera and a computer? I want to use a camera so we can agree on what we are measuring. The solution has to be cost effective. Very cost effective. Your comments will be highly appreciated Thanks
while studing ZrO2 thin film on sapphire substrate, I found a kind of modulated structure like a wave near the interface region (It's not a morie fringe). This kind of structure is very similia to the pre-martensitic sructure found in Ni-Al system. The lattice spacing in the modulated region is the same as (111) d-spacing, while in the uper region d=d(002). The film has a (001)//(0001) prefer orientation with sapphire. What I want to know is: 1. How the modulated structure formed, and how (111)//(0001) orientation can change to (001)//(0001)? Can the modulated region comes from a martensitic transformation? 2. Is there any report about modulated structure happend in ZrO2?
We have a Specimen Injection Rod and 4, PW 6145/00 3 mm. sample holders for an Philips EM 300 Biological Microscope available. Are far as I know, these have never been used because there is no evidence of contamination or wear on the rod or holders.
If you are interested, make an offer.
Fred Pearson McMaster University Brockhouse Institute for Materials Research Hamilton Ontario Canada L8S 4M1
We have a Specimen Injection Rod and 4, PW 6145/00 3 mm. sample holders tips for a Philips EM300 Biological Microscope for sale.
Are far as I know these have never been used because there is no evidence of contamination or wear on the rod or holder.
If you are interested, make an offer.
email: eoptics-at-mcmail.CIS.McMaster.CA
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
I own an old Leitz Wetzlar bi-microscope, and have no manual for maintenance, instructions, etc. Anyone here able to provide some pointers or reference? Many thanks. Frank G Anderson 745 Sipsiri Soi 3 Myang, Korat 30000 Thailand
Miguel Angel S=E1nchez wrote: } =20 } -----------------------------------------------------------------------= - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Co= m } -----------------------------------------------------------------------. } =20 } Hi there: } My name is Miguel Sanchez } my problem is the following } As part of my business I receive } micron rated textiles, that is nylon mesh } that has a determined distance between } threads. } Sometimes is very difficult to determine } that the textile is inside tolerances or } not. } I use an optical microscope to do measure them } my question is: } What do I need to measure the textiles } with the use of a camera and a computer? } I want to use a camera so we can agree } on what we are measuring. } The solution has to be cost effective. Very cost } effective. } Your comments will be highly appreciated } Thanks You need a reticule and a stage micrometer. The reticule inserts into an eyepiece and provides a grid. The stage micrometer allows for measurement of each increment.=20
You can obtain from Klarmenn Ruling, Inc. Manchester NH (603) 424-2401.
You'll need to know the ID of the eyepiece and what measurement scale you want to use.
Seth; Ted Pella offers Printerface for Windows, an image capture system that generates digital images from analog SEM images (I think the board is manufactured by Gatan). I haven't tried it yet, but I've heard it works well. (I have no financial interest in Pella or Gatan). Pella's phone # is (800) 637-3526 (USA).
Leslie Eibest Zoology Dept., Box 90325 Duke University (919) 684-2547 leibest-at-duke.edu
Sanford Simon wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Does anyone know of a source for teflon o-rings with a thickness of {100 uM? } } Thanks, } Sandy Simon } } Sanford M. Simon } Laboratory of Cellular Biophysics } Box 304 } Rockefeller University } 1230 York Avenue } New York, N.Y. 10021 } 212-327-8130 (voice) } 212-327-8022 (fax) } simon-at-rockvax.rockefeller.edu (e-mail) You can try Roger Zatkoff Company -at- (810) 478-2400, Farmington Hills, MI. They carry nothing but O Rings.
Bob Baier at the Univ of Buffalo apparently saw my name on this list, and sent me a personal message about a week ago. I have since tried several times to reply to him using the address: baier-at-ubvms.cc.buffalo.edu - (which I copied directly from the original message I received from him), but keep getting rejections due to 'local delivery' problems. Does anyone happen to know a better address for him, or have suggestions on how to get through to him with this one?
TIA
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
Chapter 3 of the book "Advanced Scanning Electron Microscopy & X-ray Micro Analysis" by Newberry, Joy, et. al, Plenum Press 1986 is entitled "Electron Channeling Contrast in the SEM", and ought to provide the information you are looking for.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
I'd be pleased to hear from people who are working with the BAL-TEC BAF-060 freeze-etch system, if nothing else, to swap stories, experiences and ideas. I'm thinking about modifying or redesigning the double replica specimen table, because the BAL-TEC (Technotrade) model looks too difficult to load without pre-fracturing the sample. Has anyone had good luck with the off-the-shelf model? or come up with a better design? The holders with 3 individual sample slots used on older Balzers systems seemed to work well, but to my knowledge that design is not available for the BAF 060.
Thanks for any feedback, Tom
Thomas H. Giddings Tel: (303) 492-8402 MCDB Electron Microscopy Service Fax: (303) 492-7744 University of Colorado giddings-at-spot.colorado.edu Boulder, CO 80309-0347
Dear Glenn, From what I can remember, the few times I have see channeling contrast on the SEM, it is what shows up when other sources of contrast are missing. The sample must be electropolished (to eliminate surface deformation), smooth (to eliminate SE contrast), homogeneous (to eliminate BS contrast), then crank the contrast way up. I think the BS detector works best. High voltage helps. Albert Curzon of the Physics Department of Simon Fraser University in Burnaby, B.C. Canada has done SEM studies of magnetic domain, which is imaged in a similar way. You wrote:
} } Can anyone refer me to a basic reference on electron channeling contrast, or failing that, give me a few hints on optimal conditions for observing this type of contrast ? } } TIA } } Glenn } } Glenn Poirier Phone (514) 398 6774 } Electron Microprobe Laboratory Fax (514) 398 4680 } Earth and planetary Science } Mcgill University Regards, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
I picked up the following message which was posted to another newsgroup (sci.techniques.microscopy). Anyone who can help, please contact J.L. Beauchamp directly at Caltech.
If you haven't noticed it the Microscopy Listserver was off-line for most of the day today.
This note is just to head off a host of messages asking why people were not able to send/receive Email and/or login to various MSA WWW sites.
Network Operations Center and Ameritek Communications had a major fault in the lines to this site. We finally came back up on-line ~ 11 PM CST . A technical genius at the main switching center basically didn't know what he was doing and incorrectly reconfigured a central router about 11 AM this morning. It took lots of phone calls to get it sorted out.
All the local systems here were running, they just could not talk to anyone on the outside world. Well not quite true, I still managed to reconfigure my backup modem connections and send a couple of messages...
AccuMed International, Inc., has an immediate opening for a technical = manager to oversee scientific and technical projects involved in the = development, scientific and clinical testing, and production of = computerized medical laboratory instruments and software-based = applications. The successful candidate will carry out day-to-day = activities involved in shepherding new instruments from R&D to finished = product. The position requires a minimum of a Bachelor's degree and a = strong scientific/technical background, with at least 3-5 years = experience in developing, marketing or supporting imaging workstations = and software-based applications for light microscopy. Specific areas of responsibility will include: - managing the development of new hardware and software products - establishing and maintaining timelines for delivery of prototype = instruments and finished products, including software-based applications - coordinating activities in various departments including R&D, = engineering, software development and manufacturing - assisting with applications to regulatory agencies
AccuMed International, headquartered in Chicago, is a global laboratory = diagnostic products company and an emerging leader in the field of = automated cytopathology. EOE.
Please respond via mail, FAX or email (NOT telephone) to:
Marc M. Friedman, Ph.D. Director, Scientifc and Technical Affairs AccuMed International, Inc. 900 N. Franklin, Suite 401 Chicago, IL 60610 Tel: (312) 642-9200 FAX: (312) 642-8684 email: marc.friedman-at-accumed.com
It is not uncommon for me to employ electron channelling contrast. Almost all of my experience is with stainless steels, nickel superalloys (Inconels), Zirconium alloys, and uranium compounds. The ecc is a weak signal and is not always easy to produce.
A few ideas....
Use BSE imaging with large beam currents and high BSE signal gain (contrast). It may be necessary let any response from areas of higher or lower Z than the matrix of interest go saturated black and/or white to achieve this.
Use a "normal" incident beam (0 degrees tilt)
Contrary to some advice I have recieved, I find that lower beam voltages (10 kv) work better than higher (20-30 kV). I have no proof, but suspect that the lower penetration depth images the surface grains without "confusing and diluting" the image with BSE returns from sub-surface grains with different orientations.
Surface preparation is VERY important. A very well polished surface, free from surface damage is required or the signal will be obscured. Some materials are easier than others to prepare. On occation, I have had to send samples back to our met-lab several times before a satisfactory surface is available. A very light "attack" polish (not really an etch) can be helpful during final polish for some materials. I have wanted to try a light ion beam cleaning, but don't have one.
Incident beam angle (with specimen at 90 nominal) changes resulting from the
raster can strongly affect contrast. Experiment with working distance vs. magnification to see what works best for you. Along this line... Don't expect to be able to generate multiple image mosaics which match well. A grain that is dark on one edge of an image may will be light when the stage is translated one frame over because the incident angle is nolonger the same.
Hope this helps...
Woody White
http://www.geocities.com/capecanaveral/3722
BTW something happened to my email pgm half way through??? Hope this dosen't "run over" line length too much!
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Good afternoon everyone.
Can anyone refer me to a basic reference on electron channeling contrast, or failing that, give me a few hints on optimal conditions for observing this type of contrast ?
TIA
Glenn
Glenn Poirier Phone (514) 398 6774 Electron Microprobe Laboratory Fax (514) 398 4680 Earth and planetary Science Mcgill University
THERE ARE THREE SIDES TO EVERY STORY: yOURS, MINE AND THE TRUTH
} Date: Tue, 25 Mar 1997 11:46:20 -0500 (EST) } From: Michael Knotts {ph281mk-at-prism.gatech.edu} } To: microscopy-at-Sparc5.Microscopy.Com } Subject: Looking for veteran SEM service engineers } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello Microscopists, } } Can anyone suggest names of service engineers with experience on } vintage Cambridge electron microscopes? I have a 1971 Cambridge } Stereoscan S4 SEM in need of service. I'd like to locate an } experienced technician in the metro Atlanta area or the southeastern } US, but I would appreciate all references and information. Thanks, } } Michael Knotts } ------------------------------------------------------------- } Michael E. Knotts, Ph.D. E-mail: ph281mk-at-prism.gatech.edu } Contributing Editor {The Light Touch} OPTICS & PHOTONICS NEWS } Georgia Tech / School of Physics / Atlanta, GA 30332-0430 } Tel: (404) 894-3422 FAX: (404) 894-9958 }
Yes, Hugh Whitaker (WHIT) used to keep our ancient Cambridge Stereoscan going with corks and paper clips from the local electronics store when he couldn't get parts from Cambridge. I think he is retired now, but he is a wealth of information, and being retired, may be available to help you out. He used to live in Raleigh, but I think he may have moved to the coast of NC. His daughter, Sharon Drew, works at Medical Univ. Hosp. in Charleston 803 792 4157. Perhaps she can get in touch with him for you. 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
} Talking of peak overlaps, is anyone out there capable of reliable } accurate analysis of V in the presence of large amounts of Ti, using: } } a WDS } } b EDS? } } I ask because I concluded recently that I couldn't, with my EDS-only } system, analyse for V in titanomagnetites, and that it probably } wasn't possible even with WDS. } Ritchie } } Ritchie Sims phone: 64 9 3737599 ext 7713 } Department of Geology fax: 64 9 3737435 } University of Auckland } Private Bag 92019 } Auckland } New Zealand
EELS on a TEM, with energy resolution around 1-2 eV, pretty easily separates Ti and V, using the L edges, and there is little problem of confusion with N K edge. Accuracy of quantitation won't be very good but it might allow you to decide if you really do have a small amount of V in Ti.
However, what is known as 'Sod's Law' ensures that the O-K edge sits almost exactly at the energy of the V-L edge! If you have strongish V (or O) edges, then you can distinguish O-K from V-L on the basis of edge shape, but if you've got that level of V, you can probably separate V and Ti by EDX anyway!!
Seth, for your optical microscope image capture, we have a program called "Image Central" that will archive and database all of your digital images and let your acquire them directly from a variety of sources. Please contact me fro further info and I will send a package of literature out to you.
Scott E. Berman Advanced Imaging Concepts, Inc. Princeton, NJ Phone: (609) 921-3629 x26 Fax: (609) 924-3010 e-mail : Scott E57-at-aol.com
I have just read an artical that includes in its protocol for fluorescent dyeing, blot drying of the samples, then drying under a stream of nitrogen.
My question was what is the benefit of drying under a stream of nitrogen as opposed to air drying? My first thought was perhaps to reduce any oxidation. Can anyone shed any light on this matter.
Thanks,
Chris. ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Chris Johns Queensland University of Technology 2 George St. Brisbane QLD 4001. Email: {c.johns-at-student.qut.edu.au} ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Please find hereafter the major characteristics of our grabbing system for SEMs: Orion 4.2 for Windows.
=B7 Digitizes in any size between 16x16 and 4096x4096 pixels (depending on your SEM) =B7 Square pixels in any mode =B7 Connects to any existing SEM or STEM =B7 100% reliable for your SEM and lowest noise level due to unique, optical isolation on board =B7 Grabs images in slow scan and full photo resolution =B7 Can grab 2 simultaneous images (for example SE and BSE fields) =B7 Unique, programmable oversampling factor dramatically reduces image= noise in real time =B7 The grabbed images are immediately available to the user in the PC RAM =B7 They are stored in full resolution, 256 grey levels per pixel (frame integration uses 16 bits per pixel during grabbing) =B7 Easy transfer to any other application (OLE 2 or clipboard) =B7 Distance measurement, extraction, filtering, line / rectangle drawings, line scan function =B7 Selectable, enhanced image compression engine reduces image size on disk by 90% with little image degradation =B7 Full compatible with Windows 3.1x, Windows 95 and Windows NT.
Options =B7 EDX mapping grabbing mode mixes elemental maps with SE or BSE image =B7 Ultra high resolution photo replay allows image to be photographed later from disk =B7 HP Laserjet enhancement card allows photographic quality in 10 seconds =B7 Powerful yet simple programming language makes the user=92s job easier
See also our WEB site http://www.microscopy-uk.org.uk
At 16:33 24/03/1997 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
Best regards,
Paul Vanderlinden. Sales Manager.
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D See our web site: http://www.microscopy-uk.org.uk =20
} } Does anyone know of a source for teflon o-rings with a thickness of {100 uM? } } Thanks, } Sandy Simon } } Sanford M. Simon } Laboratory of Cellular Biophysics } Box 304 } Rockefeller University } 1230 York Avenue } New York, N.Y. 10021 } 212-327-8130 (voice) } 212-327-8022 (fax) } simon-at-rockvax.rockefeller.edu (e-mail)
We purchased teflon o-ring 9 years ago from Bal Seal Engineerig Co., 620 W. Warner Ave. Santa Ana, CA 92707-3398. Tel: 714-557-5192.
Ya Chen
Ya Chen
========================================================================= \ / Integrated Microscopy Resource (IMR)-- \ / __ an NIH Biomedical Research Resource TEL : 608-263-8481 \/ / / University of Wisconsin-Madison FAX : 608-265-4076 / / / 1675 Observatory Drive #159 Email1:ychen14-at-facstaff.wisc.edu / /__/_ Madison, WI 53706 Email2:chen-at-calshp.cals.wisc.edu ========================================================================= IMR WWW Home Page: http://www.bocklabs.wisc.edu/imr.html
I request your help in locating a publisher or bookseller who carries the book "Practical electron microscopy in Material Science" by J.W.Edington. It was originally published as a 4 vol. series by Philips. Van Nostrand who published the combined volume in 1976 say the book is out of print.
Thanks for your help.
Mohan Kalyanaraman Sr. Staff Material Scientist Mobil Technology Company Paulsboro, NJ 08066 609-224-3989
Chapter 3 of the book "Advanced Scanning Electron Microscopy & X-ray Micro Analysis" by Newberry, Joy, et. al, Plenum Press 1986 is entitled "Electron Channeling Contrast in the SEM", and ought to provide the information you are looking for.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
Bob Baier at the Univ of Buffalo apparently saw my name on this list, and sent me a personal message about a week ago. I have since tried several times to reply to him using the address: baier-at-ubvms.cc.buffalo.edu - (which I copied directly from the original message I received from him), but keep getting rejections due to 'local delivery' problems. Does anyone happen to know a better address for him, or have suggestions on how to get through to him with this one?
TIA
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
There are a few vendor that offer Image capturing for the SEM. Most of the system we have installed on our refurbished SEM are supplied by Evex Analytical. There number is 609-252-9192.
I am trying to find out what is the resolution of a Confocal Scanning Laser Microscope. Does it depend on the type of laser used?
I also have another question concerning SEM microscopes. Why is the column of the SEM shorter than the TEM. It is because of a simpler electron optic system in the SEM?.
I know that these are basic questions, but answers to questions like these are impossible to find in textbooks
} Dear All, } } I request your help in locating a publisher or bookseller who carries } the book "Practical electron microscopy in Material Science" by } J.W.Edington. It was originally published as a 4 vol. series by } Philips. Van Nostrand who published the combined volume in 1976 say } the book is out of print. } } Thanks for your help. } } Mohan Kalyanaraman } Sr. Staff Material Scientist } Mobil Technology Company } Paulsboro, NJ 08066 } 609-224-3989
Henk Colijn colijn.1-at-osu.edu OSU Campus Electron Optics Facility ------------------------------------------------------------------- Pereant qui ante nos nostra dexerunt.
Hope I am not double posting... The orginal didn't show up and may have been lost is cyberspace...??? _W_
It is not uncommon for me to employ electron channelling contrast. Almost all of my experience is with stainless steels, nickel superalloys (Inconels), Zirconium alloys, and uranium compounds. The ecc is a weak signal and is not always easy to produce.
A few ideas....
Use BSE imaging with large beam currents and high BSE signal gain (contrast). It may be necessary let any response from areas of higher or lower Z than the matrix of interest go saturated black and/or white to achieve this.
Use a "normal" incident beam (0 degrees tilt)
Contrary to some advice I have recieved, I find that lower beam voltages (10 kv) work better than higher (20-30 kV). I have no proof, but suspect that the lower penetration depth images the surface grains without "confusing and diluting" the image with BSE returns from sub-surface grains with different orientations.
Surface preparation is VERY important. A very well polished surface, free from surface damage is required or the signal will be obscured. Some materials are easier than others to prepare. On occation, I have had to send samples back to our met-lab several times before a satisfactory surface is available. A very light "attack" polish (not really an etch) can be helpful during final polish for some materials. I have wanted to try a light ion beam cleaning, but don't have one.
Incident beam angle (with specimen at 90 nominal) changes resulting from the
raster can strongly affect contrast. Experiment with working distance vs. magnification to see what works best for you. Along this line... Don't expect to be able to generate multiple image mosaics which match well. A grain that is dark on one edge of an image may will be light when the stage is translated one frame over because the incident angle is nolonger the same.
Hope this helps...
Woody White
http://www.geocities.com/capecanaveral/3722
BTW something happened to my email pgm half way through??? Hope this dosen't "run over" line length too much!
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Good afternoon everyone.
Can anyone refer me to a basic reference on electron channeling contrast, or failing that, give me a few hints on optimal conditions for observing this type of contrast ?
TIA
Glenn
Glenn Poirier Phone (514) 398 6774 Electron Microprobe Laboratory Fax (514) 398 4680 Earth and planetary Science Mcgill University
THERE ARE THREE SIDES TO EVERY STORY: yOURS, MINE AND THE TRUTH
} } I also have another question concerning SEM microscopes. Why is the } column of the SEM shorter than the TEM. It is because of a simpler } electron optic system in the SEM?. } } I know that these are basic questions, but answers to questions like } these are impossible to find in textbooks } } Thanks. } } GCH.
Hi Gary,
Basicly, a TEM has condenser, objective, intermediate, and projection lenses, plus a fluorescent screen and a film chamber. In contrast, SEM has only condenser and objective lenses, plus a speciemen chamber. That is obviously why SEM has a short column.
Ya Chen
Ya Chen
========================================================================= \ / Integrated Microscopy Resource (IMR)-- \ / __ an NIH Biomedical Research Resource TEL : 608-263-8481 \/ / / University of Wisconsin-Madison FAX : 608-265-4076 / / / 1675 Observatory Drive #159 Email1:ychen14-at-facstaff.wisc.edu / /__/_ Madison, WI 53706 Email2:chen-at-calshp.cals.wisc.edu ========================================================================= IMR WWW Home Page: http://www.bocklabs.wisc.edu/imr.html
The Integrated Microscopy Resource and Carnegie Mellon University will be sponsoring a symposium and short course on multi-photon excitation imaging, August 9-10, 1997, in Cleveland Ohio.
} I ask because I concluded recently that I couldn't, with my EDS-only } system, analyse for V in titanomagnetites, and that it probably } wasn't possible even with WDS. } If I ask my system to look for V in my standard Rutile (TiO2), it } always finds a few tenths of a %, even though my software } (LINK ZAF4/FLS) strips out Ti Kb along with the Ka. I have no way of } telling whether this V is genuine or not, but I suspect that it } isn't, as my supposedly-pure Ti metal standard also gets credited } with a small amount of V, which I doubt. } I was recently given another rutile standard, and, blow me down, it } is reputed to have 0.4% V by WDS analysis. } Anyone got any feeling for whether this is likely to be true?
The general case for interference with the transition elements is that the Z-1 element Kb peak overlaps the Z element Ka peak. Ti Kb on V Ka is what you bring up.
For EDS analysis, assuming that linear least squares deconvolution is used, the fit for Ti should handle both Ti Ka and Kb if they are grouped together as your EDS reference. Any residual could then appear as a positive value for V. This may reflect a calibration difference between your sample and reference spectra (I'm assuming we are not talking standardless EDS here).
For WDS measurement one has the choice of PET or LIF; the latter has better resolution and should be used to reduce the magnitude of the overlap to begin with. I find that (using LIF) the measured V Ka k-ratio on synthetic (pure) TiO2 is typically less than 0.5%, which compares with:
} I was recently given another rutile standard, and, blow me down, it } is reputed to have 0.4% V by WDS analysis.
My guess is that the analyst did not correct for the overlap. In the absence of any more sophisticated software, you can make a correction for a simple overlap like this via:
corrected V K-ratio = measured V K-ratio - AK * measured Ti K-ratio
where AK is the apparent V K-ratio measured on pure TiO2. Thus, when analyzing TiO2 the resulting V concentration is zero, and when analyzing a phase containing no Ti the correction is zero. This method corrects the K-ratio before passing to the ZAF program, but you could just as well make a small correction like this on the weight percent data. In practice I find that this linear correction reduces the apparent V in pure TiO2 by an order of magnitude, i.e. from 0.X% to 0.0X%, so especially several hundred ppm V in rutile should be viewed with scepticism.
You can also do a wavelength scan on your rutile looking for the presence of the V *Kb* peak, since this is not overlapped. If you see this peak, then you know there is V in the sample.
This is not a bad overlap. Consider Pb La on As Ka using LIF. This is a total overlap with no real possible solution via this linear correction. The solution in a case like this is to use As Kb and either Pb Lb or Pb Ma as your lines (the application is for sulfides or arsenides). As long as there are no absorption edges between the Ka and Kb lines, or La and Lb lines, the mass absorption coeffiecients are usable; one really has no other choice.
Have fun,
Paul
+----------------------------------------------------+ | Paul K. Carpenter paulc-at-gps.caltech.edu | | Division Analytical Facility | | Geological and Planetary Sciences MC 100-23 | | California Institute of Technology | | Pasadena, CA 91125 | | 818-395-6126 (X-ray Lab) 818-568-0935 (Dept. FAX) | +----------------------------------------------------+
} I have just read an artical that includes in its protocol for fluorescent } dyeing, blot drying of the samples, then drying under a stream of nitrogen. } } My question was what is the benefit of drying under a stream of nitrogen as } opposed to air drying? My first thought was perhaps to reduce any oxidation.
Oxidation is one consideration. The lack of water vapor in the N2 stream is another, and lack of dust, oil droplets, etc. in N2--these are often present in compressed air--is a third.
} Can anyone shed any light on this matter. } If I did, would you just send it back longer? ;-) Yours, Bill Tivol
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
My LEO service engineer recommended that I contact you to get an e-mail address for Chris Martinic at the University of New South Wales. Chris has information about image grabbers that I need. Please send me his e-mail address. My address is:
tom.cox-at-lmco.com
Thank you,
Thomas J. Cox Lockheed Martin Missiles & Space Co.
I would like to know what goes on inside the chamber of my electron microprobe when the focused, moderate to high current (10 - 200 nA) electron beam hits epoxy. Visually, a crater forms. What are the solid/gaseous by-products of that reaction? Any ideas, educated guesses, and/or experimental or theoretical results (or suggestions where to look) would be appreciated. Thanks.
John
John Fournelle Electron Microprobe Lab office: (608) 262-7964 Dept of Geology & Geophysics fax: (608) 262-0693 University of Wisconsin home: (608) 274-2245 1215 West Dayton St. email: johnf-at-geology.wisc.edu Madison, WI 53706 amateur radio: WA3BTA/9 Personal http://geology.wisc.edu/~johnf/ Probe lab http://geology.wisc.edu/~johnf/sx51.html
"The first rule of all intelligent tinkering is to save every cog and wheel." Aldo Leopold
} I am trying to find out what is the resolution of a Confocal Scanning } Laser Microscope. Does it depend on the type of laser used? } Dear Gary, The resolution of a CSLM is ~1/3 to ~1/2 the wavelength of the ilumination in the x-y plane and about 2x worse in the z-direction. Jim Pawley's Handbook of Confocal Microscopy has a very good description of the theory. (I hope I got the title right--the book is at my desk & I'm not.) It will depend on the wavelength of light used, but not on whether the laser is pulsed or continuous wave, so whether it depends on the "type of laser used" depends on what you mean. There may be CSLM's which use frequency-doubled light, so a pulsed laser is neces- sary to provide sufficient intensity to get the non-linear optics to work, so if this is what you mean, then whether the CSLM works at all can depend on the type of laser used, but once you get light of a par- ticular wavelength, the resolution won't depend on where that light came from. Since I'm not an expert on this, you should probably check the book. Yours, Bill Tivol
} } Hello Microscopists, } } } } Can anyone suggest names of service engineers with experience on } } vintage Cambridge electron microscopes? I have a 1971 Cambridge } } Stereoscan S4 SEM in need of service. I'd like to locate an } } experienced technician in the metro Atlanta area or the southeastern } } US, but I would appreciate all references and information. Thanks,
Clark Houghton at Secondary Images is a real wizard with the old Cambridges. He is in Ohio (but was willing to come all the way to Hawaii... what a hardship!) at (513) 927-5373.
Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Hello! We have a Cambridge Stereoscan and for service use Scanners Corporation in Eldersburg, MD - tel. 1-410-549-3800 I believe they cover the East Coast and possibly a bit further out as well. They take great care of our SEM. Nancy
In response to the question as to why SEM colums are so much smaller than those of TEMs, I offer the following:
The colums of SEMs basically contain only two lenses: a 'first' lens (often incorrectly called the condenser lens) which produces demagnification of the spot from the electron gun, and a 'second' lens (usually incorrectly called the objective lens) which is used to focus the spot image produced by the first lens onto the specimen forming the electron probe which generates the signals that are emitted from the sample. Sometimes the first lens is a compound lens, actually giving a total of three lenses; nonetheless, they function as a two-lens system overall.
This entire electron optical system of an SEM corresponds in general character and function to the 'condenser lens system' (i.e. the system of lenses that is above the specimen chamber) of a TEM. That is, the condenser lens system of a TEM is basically a two-lens system which functions to control the illumination that strikes the specomen (just as the two lens system of the SEM does).
In a TEM; however,there is an additional image-forming lens system below the specimen consisting of the objective lens, which produces the primary electron image from the specimen, and then a series of from 3 to 6 additional lenses which produce the wide range of magnifications we expect from TEMs now-a-days, plus providing the variety of additional imaging functions commonly available (selected area diffraction, convergent beam diffraction, etc.). No lens is directly involved in the image-forming process in an SEM, and so these additional lenses are not needed.
Thus there are three or four times as many lenses in a TEM as there are in an SEM. In addition, whereas most TEMs are designed to operate at electron accelerating voltages from 80 kV to several hundred kilovolts, most SEMs do not use accelerating voltages much above 30 kV. Since it is much easier to deflect these lower energy electrons, the magnetic fields in SEM lenses do not need to be as strong as those in TEMs; consequently they require fewer turns of wire in their excitation coils, and so can be physically much smaller.
I hope this answers your question satisfactorily. If not, don't hesitate to let me know. I can send you column diagrams which illustrate the above points, if needed.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
In response to the question as to why SEM colums are so much smaller than those of TEMs, I offer the following:
The colums of SEMs basically contain only two lenses: a 'first' lens (often incorrectly called the condenser lens) which produces demagnification of the spot from the electron gun, and a 'second' lens (usually incorrectly called the objective lens) which is used to focus the spot image produced by the first lens onto the specimen forming the electron probe which generates the signals that are emitted from the sample. Sometimes the first lens is a compound lens, actually giving a total of three lenses; nonetheless, they function as a two-lens system overall.
This entire electron optical system of an SEM corresponds in general character and function to the 'condenser lens system' (i.e. the system of lenses that is above the specimen chamber) of a TEM. That is, the condenser lens system of a TEM is basically a two-lens system which functions to control the illumination that strikes the specomen (just as the two lens system of the SEM does).
In a TEM; however,there is an additional image-forming lens system below the specimen consisting of the objective lens, which produces the primary electron image from the specimen, and then a series of from 3 to 6 additional lenses which produce the wide range of magnifications we expect from TEMs now-a-days, plus providing the variety of additional imaging functions commonly available (selected area diffraction, convergent beam diffraction, etc.). No lens is directly involved in the image-forming process in an SEM, and so these additional lenses are not needed.
Thus there are three or four times as many lenses in a TEM as there are in an SEM. In addition, whereas most TEMs are designed to operate at electron accelerating voltages from 80 kV to several hundred kilovolts, most SEMs do not use accelerating voltages much above 30 kV. Since it is much easier to deflect these lower energy electrons, the magnetic fields in SEM lenses do not need to be as strong as those in TEMs; consequently they require fewer turns of wire in their excitation coils, and so can be physically much smaller.
I hope this answers your question satisfactorily. If not, don't hesitate to let me know. I can send you column diagrams which illustrate the above points, if needed.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Not my field, but wavelength of illumination will be one determining factor.
} I also have another question concerning SEM microscopes. Why is the } column of the SEM shorter than the TEM. It is because of a simpler } electron optic system in the SEM?.
Two answers:
1. Voltage - generally, SEMs work at a maximum of 30kV while TEMs typically work from a minimum of 100kV. Higher voltage electron have much greater kinetic energy and are thus more diffcult to deflect. Even with somewhat stronger lenses, the additional energy of the electron requires a little more distance to have the required effect.
2. More significantly, an SEM column is only equivalent to the part of the TEM colum ABOVE the specimen. In a TEM, additional lenses are needed after the specimen to focus the beam into an image. In a SEM, the electron beam is aleady focused to a probe at the specimen, and only needs 'intensity' detectors. Indeed, if you do secondary electron imaging on a bulk specimen in a TEM/STEM type if instrument, the bottom half of the column is not used.
} I know that these are basic questions, but answers to questions like } these are impossible to find in textbooks } } Thanks. } } GCH.
Regards,
--------------------------------------------------------------- Dr L. P. Stoter Technical Editor, MICROSCOPY & ANALYSIS Technesis 17, Rocks Park Road email: LPS-at-teknesis.demon.co.uk Uckfield, E. Sussex Phone: +44 (0)1825 766911 TN22 2AT Fax: +44 (0)1825 766911 United Kingdom ---------------------------------------------------------------
Seth: we built an SEM image-capture system based on a National Instruments (AT-MIO-16E-10) data-acquisition card, controlled by a program written in Visual Basic. Hardware cost was about $US 3000 including Pentium computer and 1200 dpi laser printer. Brief details of the system will be presented at the June MSC conference, 2-page abstract appearing soon on the conference web page: http://www.ualberta.ca/~mmid/msc
Ray Egerton, Physics Dept, University of Alberta, Edmonton, Canada T6G 2J1 Phone: 403-492-5095, FAX: 403-492-0714, e-mail: egerton-at-phys.ualberta.ca ------------------------------------------------------------------------
I have not had much experience with scanners, but it seem to me the answer to why there columns are not as long as the TEM's are maybe in the fact that you do not have the objective len and stage or the projector or diffraction lens to deal with. Thats my best guess. As for Confocal Scanning I have had no experience at all with this device. sorry!
Dear John, When I did a careful study of SEM sample contamination, using a polished copper sample and carefully regulated kV, current and magnification, the deciding factor seemed to be how recently someone had hit epoxy with the beam. One "hit" seemed to result in much worse contamination for about three days. In high current situations you can see the epoxy "boil". I think the vapourised hydrocarbons fly about your chamber for quite a while. Cleaning with a N2 purge, like the SEMclean system (I use a home-built one), does seem to help. I seem to recall that someone did a study of contamination, air-jets and such in Microbeam Analysis in the early '80s. May have been Bastin. You wrote:
} I would like to know what goes on inside the chamber of my electron microprobe } when the focused, moderate to high current (10 - 200 nA) electron beam hits } epoxy. Visually, a crater forms. What are the solid/gaseous by-products of } that reaction? Any ideas, educated guesses, and/or experimental or } theoretical results (or suggestions where to look) would be appreciated. } Thanks. } } John Hope this helps, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
You might want to check out 4Pi Analysis. they have a a Mac-based image capture system for SEMs that is priced very competitively. There boards and software offer total control over the SEM, with image acquistion by NIH Image, Photoshop, IPLab Spectrum, etc.
Regards, Glen MacDonald Virginia Bloedel Hearing Research Center Box 35-7923 University of Washington Seattle, WA 98195-7923 (206) 616-4156 glenmac-at-u.washington.edu
/*---------------------------------------------------------------------- -*/ / The box said "Requires Windows 95 or better.", so I bought a Macintosh. / /*---------------------------------------------------------------------- -*/
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Subject: Time:9:50 AM OFFICE MEMO In Memoriam - Mark Fendorf Date:3/28/97
In Memoriam - Mark Fendorf
It is with great sadness that we at NCEM announce the death of Mark Fendorf.
Mark John Fendorf died on March 14, 1997 at the age of 36. As a collaborative post-doctoral scientist at LBNL's National Center for Electron Microscopy, Mark played a central role in NCEM's outreach to external facility users. He was an outstanding microscopist, a resourceful materials scientist and a delightful colleague to all members of NCEM. In his role of resident expert microscopist he collaborated with many NCEM users and helped countless visitors of the facility. He earned his Ph.D. in materials science from UC Berkeley in 1992. His scientific work focused on electron beam micro-characterization of a number of different materials, including high-temperature superconductors, catalysts, fullerines and most recently the adsorption of heavy metals on soil minerals. He was an active member of several scientific societies, including the Microscopy Society of America and the American Ceramic Society. In 1991, he co-founded the UC Berkeley chapter of the Materials Research Society. Outside of LBNL, he was a dedicated and enthusiastic volunteer for the Boy Scouts of America where he was a leader for 16 years, served on the Monterey Bay Area Council, and acted as Director of Troop Leadership Training for a number of years.
During his progressively debilitating illness, Mark continued his work at NCEM with remarkable dedication and tenacity. His strength of will and his determination to overcome his handicap caused by the illness were admired by all those who knew him. Mark's untimely death has left the Center permanently diminished. He will be greatly missed by his NCEM colleagues and the many users and visiting scientists at the facility.
Mark Fendorf is survived by his parents, Ken and Virginia Fendorf and his brothers Scott and Dale. In recognition of his sustaining enthusiasm for the science of electron microscopy, and in an effort to help others continue where Mark had to leave off, his family has established a memorial fund in support of NCEM's outreach program.
Donations to the Mark Fendorf memorial fund can be made to: World Savings, Aptos Branch, Mark Fendorf Memorial Fund, 7970 Soquel Drive, Aptos, CA 95003
} I would like to know what goes on inside the chamber of my electron microprobe } when the focused, moderate to high current (10 - 200 nA) electron beam hits } epoxy.
When an electron passes through matter, the energy of the electron is transferred to the material in the form of ionizations and excitations. About 30 eV is transferred for each ionization, and most of the energy transferred goes into primary or secondary ionizations. The products, ob- viously far from thermodynamic equilibrium, react with other components of the material causing chemical bond breaking and free radical formation. Finally (a few microseconds later) these reactive species form more stable products, which are often small organic molecules in the case of electrons incident on epoxy resin. There is a statistical distribution of products which depends on the nature of the resin; since many of these are volatile, they will travel throughout the chamber (they also gain some kinetic energy from the energy transferred from the initial electron).
} Visually, a crater forms.
This should make sense in light of the above.
} What are the solid/gaseous by-products of } that reaction? Any ideas, educated guesses,
The above are educated guesses...
} and/or experimental or } theoretical results
and theoretical results.
} (or suggestions where to look) would be appreciated.
My info comes from Friedlander, et al., Nuclear and Radiochemistry; I do *not* recommend this book. A good book on radiation chemistry would be a good place to start, but I don't know any titles. Yours, Bill Tivol
Jeol sells a high resolution standard (gold on carbon) that we would like to purchase. Does anyone have info: (part number, price, where to order)? Many thanks.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
A formal proposal to create a new group tentatively called sci.bio.immunocytochem was posted to news.announce.newgroups on 17.3.97. This is a reposting of that proposal. Please have a read and let me know what you think.
REQUEST FOR DISCUSSION (RFD) unmoderated group sci.bio.immunocytochem
This is the 2nd Request For Discussion (RFD) for the creation of a world-wide unmoderated Usenet newsgroup sci.bio.immunocytochem, currently being discussed in news.groups
Suggestions for improvements to this proposal are welcome. Discussion about it should take place in news.groups. A vote is expected to be held in about three to four weeks.
This is not a Call for Votes (CFV); you cannot vote at this time. Procedural details are below.
CHANGES from previous RFD:
2nd RFD posted because more than 60 days have elapsed since 1st RFD. There are (minor changes in Distribution and Newsgroup line.
Newsgroup line: sci.bio.immunocytochem Immuno-labelling of biological material.
RATIONALE: sci.bio.immunocytochem
Immunohistochemists and immunocytochemists already enjoy the benefits of online communication, utilizing e-mail, accessing web sites, and subscribing to specialised mailing lists. Usenet newsgroups are also popular, but this is less obvious because articles with immunocytochemical/immunohistochemical content get posted to many different newsgroups. Most articles are posted to a favourite five or six newsgroups including bionet.cellbiol, sci.med.immunology and sci.techniques.microscopy, but often articles get posted to any one of fourteen or fifteen newsgroups in the sci. and bionet. heirarchies. Some of these are listed in the distribution list at the end of this proposal.
In my view, no existing newsgroup fulfils the criteria necessary to attract all the various immunocytochemistry postings. I do not wish to draw users away from other newsgroups, only to encourage scientists to share their knowledge and expertise on immunocytochemistry in the most effective manner. In response to my proposal to create a newsgroup dedicated to the discussion of immunocytochemistry and immunohistochemistry, I have received e-mail and faxes from researchers all over the world offering their support and encouragment.
Immunocytochemistry and immunohistochemistry are not subdivisions of immunology, molecular biology or chemistry. Microscopy, although essential, is only a small part of the story. Immunocytochemistry and immunohistochemistry are multi- disciplinary, therefore discussions are destined to stay distributed amongst the different newsgroups until they are all brought together under one umbrella. This would then act as a focus point for all the immunocytochemists who are already Internet users, and encourage new subscribers to Usenet.
CHARTER: sci.bio.immunocytochem
This is a newsgroup for the exchange of information relating to immunocytochemistry and immunohistochemistry. This unique research tool is used to locate and identify specific molecules in biological material, at the microscopical level.
Articles posted to this group must be relevant to one or more aspects of the above. The kind of subjects that may be discussed include techniques, theory, presentation of results, requests for collaboration, history, equipment, publication references, notice of events, tips and trouble-shooting, jobs offered andwanted, jokes, stories and new ideas, so long as the posting bears a direct relevance to the central theme. There will be a list of Frequently Asked Questions (FAQs) to help newcomers.
A relevant posting could just be a simple question or answer, for example "Has anyone got any experience with this reagent ?"or "Which course could I attend to learn more about immunogold labelling?". There will be articles reminding people to read the list of FAQs prior to posting their own article. Usenet readers may get involved in complex discussions about, for example, multiple labelling, proper use of control experiments, microwave antigen retrieval or quantitative measurements. Remember that articles posted to a newsgroup are intended for a wide readership, so if you have information which concerns only one or two people then please don't use this newsgroup, use e-mail.
Commercial advertisements for services, equipment or reagents violate the charter unless one or more of the following apply: (a)The advertisement is part of a comprehensive article designed specifically to address issues raised in earlier articles posted to the group (b)A general reference to the type of product does not suffice for technical reasons and it is necessary to specify the exact commercial product (c) The information is offered primarily for the benefit of the readers (d)The advertisement is for second-hand equipment specific to immunocytochemistry (e) Requests or offers for free products are acceptable if they are not part of a sales promotion.
END CHARTER.
PROCEDURE:
This is a request for discussion, not a call for votes. In this phase of the process, any potential problems with the proposed newsgroups should be raised and resolved. The discussion period will continue for a minimum of 21 days (starting from when the 2nd RFD for this proposal is posted to news.announce.newgroups), after which a Call For Votes (CFV) may be posted by a neutral vote taker if the discussion warrants it. Please do not attempt to vote until this happens.
All discussion of this proposal should be posted to news.groups. This RFD attempts to comply fully with the Usenet newsgroup creation guidelines outlined in "How to Create a New Usenet Newsgroup" and "How to Format and Submit a New Group Proposal". Please refer to these documents (available in news.announce.newgroups) if you have any questions about the process.
DISTRIBUTION:
This RFD has been posted to the following newsgroups: news.announce.newgroups,news.groups,bionet.cellbiol, bionet.diagnostics,bionet.immunology,bionet.microbiology, bionet.molbio.methds-reagnts,sci.bio.misc,sci.med.immunology, sci.techniques.microscopy
This RFD will be reposted to the following newsgroups after its posting in news.announce.newgroups: bionet.molbio.proteins,bionet.neuroscience,bionet.plants, sci.bio.microbiology,sci.med,sci.med.laboratory,sci.misc, sci.nanotech
This RFD will also be reposted to the following mailing lists after its posting in news.announce.newgroups:
Histonet mailing list: {histonet-at-pathology.swmed.edu} Information pertaining to the technical aspects of histology and histopathology such as tissue fixatives and processing, routine histology, special stains, immunohistochemistry, in-situ hybridization etc. To subscribe type "subscribe digest" into the subject box and leave the text box empty, or to subscribe to the full service just type "subscribe". For more info access web site http://www.mwrn.com/subject/histonet.htms
Microscopy Society of America listserver: Questions/comments/answers in the various fields of Microscopy Currently over 3000 subscribers. To subscribe send the message "subscribe" to {Listserver-at-MSA.Microscopy.Com} then send messages in plain text to {Microscopy-at-MSA.Microscopy.Com} For more info access web site http://www.amc.anl.gov/ Docs/anl/Nestor/Software/telecommList.html
Stanford University list server To subscribe, send a message to {majordomo-at-pathology.stanford.edu} with "subscribe ipox-l" in the body of your message. This list helps pathologists and other laboratory professionals to exchange information about immunoperoxidase methods.
This RFD will also be reposted to the following web-sites after its posting in news.announce.newgroups:
Royal Microscope Society http://www.rms.org.uk Web Master Dr R. A. D. Mackenzie {r.a.mackenzie-at-open.ac.uk}
Center for Cell Imaging Department of Cell Biology Yale University School of Medicine Introduction to Immunocytochemistry http://info.med.yale.edu/cellimg/CCIimmuno.html Web Master Paul Webster { paul_webster-at-yale.edu}
Proponent: Amanda Wilson {awilson-at-aw.u-net.com} Proponent: Paul Monaghan { monaghan-at-icr.ac.uk} Mentor: Jonathan Grobe {grobe-at-netins.net}
simple question, when recharging a balzers gun with platinum, where should the tip of the carbon rod holding the platinum pellet be with repect to the tungsten coil?
Tx
simon
-- ------------------------------------------------- Simon C. Watkins Ph.D Associate Professor Director Center for Biologic Imaging University of Pittsburgh Pittsburgh PA 15261 tel 412-648-3051 fax 412-648-2004 -----------------------------------------------
I'm about to attempt UV polymerization of Unicryl for the first time and need advice about lamps, embedding molds which are UV penetrable, etc., etc. Virtually no specific advice is given in the circular I received or in the two papers referenced in this literature. I would appreciate any tips, and/or references from any experienced user. Thank you. Grace Kennedy
Luc Nocente wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Try Visilog from Noesis Vision Inc, we have a turn key system available for } sperm analysis and or fibre analysis. You can reach us at } http://www.noesisvision.com } } At 12:51 PM 3/7/97 +0530, SONEJA A K wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } -----------------------------------------------------------------------. } } } } } } Hello guys, } } } } Looking for image analysis software/solutions for the following: } } } } 1} sperm analysis,cell motility etc. } } 2} textile analysis,rayon fibre analysis,broken fibre etc. } } } } Could anyone point out some good manufacturers/sources with } } address/email/fax wwith name of source.?////// } } I would be grateful if someone could help me in this context. } } } } Thanks, } } } } Best regards, } } } } Anish } } } } ************************************************************************* } } For further details please contact: } } Soneja A.K. } } Director } } METZER BIOMEDICAL & ELECTRONICS PVT.LTD. } } 327 Wadala Udyog Bhavan,Wadala,MUMBAI(BOMBAY )400 031.INDIA } } Tel:91 22 4145057/4165650 } } Fax 91 22 4168757 } } } } Email:soneja-at-giasbma.vsnl.net.in } } ************************************************************************* } } } } } } ---------------------------------------------------------------------------- } --------------------- } Luc Nocente Tel: 514 345 1400 } Noesis Vision Inc. Fax: 514 345 1575 } e-mail: ln-at-noesisvision.com } 6800 Cote de Liesse, Suite 200 } St-Laurent, PQ } H4T 2A7,Canada } } Visit our new web site at http://www.noesisvision.com } ---------------------------------------------------------------------------- } ---------------------Dear Anish,
A belated response to your message of 3/7 re: image analysis for sperm motility/analysis:
We did some work recently with Hamilton-Thorne Research. They have several interesting, built-for-purpose systems. Contact: Dr. Dairmid-Douglas Hamilton 100 Cummings Place, Suite 102C 181 Elliott Street Beverly, MA 01915 Phone: (508)921-2050 Fax: (508)921-0250 Please feel free to mention my name.
John- My JEOL parts catalog lists a gold on carbon standard. The part # is 613149. Orders can be sent to JEOL (USA) Inc., Parts Dept., 11 Dearborn Road, Peabody, MA 01960, or call (508) 535-5900 and ask for parts. I don't have a recent price list, so I can't help you there. Leslie
Leslie Eibest Zoology Dept., Box 90325 Duke University (919) 684-2547 leibest-at-duke.edu
We have used a turbo pumped SEM since 1986. Our experience with this system has actually been quite good - we have only had to replace it once. It was fortunate that we are under contract because the pump cost is rather high. The greatest benefits that I see from this system are 1) pump-down time (usually only a few minutes) and 2) cleanliness. One other thing to consider; on our system, the turbo hangs down from a metal bellows connected to the base of the chamber. At one time, we had a very small crack in this bellows, which was not detected for over a year. We replaced numerous ion pumps and other components until it was finally located. Again, fortunately we have always had the system under contract.
-Bob ******************************** Bob Citron Chiron Vision 555 W. Arrow Hwy Claremont, CA 91711 PH: (909)399-1311 Email: Bob_Citron-at-cc.chiron.com ********************************
------------------------------------------------------------------------=20 The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20 John Bozzola wrote: =20 Jeol sells a high resolution standard (gold on carbon) that we would like=20 to purchase. Does anyone have info: (part number, price, where to order)?=20 Many thanks. =20 ####################################################################=20 John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 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=20 #################################################################### =20 John; You can contact JEOL parts directly in Peabody, MA or if you have the=20 right lab equipment you can make your own. A vacuum evaporator and a=20 30-40 amp. external power supply that can be connected to a second set= =20 of evaporator electrodes is all the equipment that is required. The=20 supplies are standard EM supplies that can ordered from any of the=20 usual sources. If you are interested in making your own I'd be glad=20 to send more details on/off line. =20 John Humenansky Braun Intertec Corp. 6875 Washington Ave. So. Minneapolis, MN 55439 (612) 942-4844 =20
If you have a sputter coater or evaporator, why not make your own. I've had good luck making calibration standards like this (I use them for camera constant calculation, resolution, demos, in TEM). It's good practice in thin film making for students, too ;-)
1. Make a thin carbon film by carbon coating a formvar, butvar etc. coated grid. (how thin the carbon should be depends on the final use; if you want to see the gold lattice in TEM it should be painfully thin). Remove the plastic backing by placing it in a (glass) Petri dish lined with a filter paper pad soaked in CHCl3 (hood).
2. After an overnight stint in the Petri dish, you should have mostly carbon left on the grids.
3. Pop them in your sputter-coater and give them a short blast of Au (we've got a pure Au target) if you use Au/Pd in yours, I'm not sure what you'll get... Well, if you've got a gold target about 1/12 of your normal sputtering time to coat an average SEM sample should work. You'll have a collection of isolated islets of Au. Carbon tape it to a stub and have a look.
Hope this helps, even though it doesn't answer the question.
cheers,
John Heckman TEM supervisor/Center for Electron Optics Michigan State University
Disclaimer: The preceding technique works in my reality; follow my instructions at your own risk!
} -----------------------------------------------------------------------. } } Jeol sells a high resolution standard (gold on carbon) that we would like } to purchase. Does anyone have info: (part number, price, where to order)? } Many thanks. } } #################################################################### } John J. Bozzola, Ph.D., Director } Center for Electron Microscopy } Neckers Building, Room 146 - B Wing } Southern Illinois University } Carbondale, IL 62901-4402 } U.S.A. } Phone: 618-453-3730 } Fax: 618-453-2665 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/departments/shops/cem.html } ####################################################################
Hi folks We have a em300 that is about to join the great trashcan in the sky. Before it meets its demise I was wondering whether anyone wants parts from it,
let me know
simon
-- ------------------------------------------------- Simon C. Watkins Ph.D Associate Professor Director Center for Biologic Imaging University of Pittsburgh Pittsburgh PA 15261 tel 412-648-3051 fax 412-648-2004 -----------------------------------------------
I would appriciate hearing from anyone on/off line with experience=20 with turbo pumped SEM's (benefits, horror stories, etc.) =20 Thank you =20 John Humenansky Braun Interec Corp. 6875 Washington Ave. So. Minneapolis, MN 55439 (612) 942-4822
Jeol sells a high resolution standard (gold on carbon) that we would like to purchase. Does anyone have info: (part number, price, where to order)? Many thanks.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 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 resolution standard can be purchased from the JEOL USA parts department. Phone # (508)535-5900. The part number is 613149. I believe this is what you are asking for but verify that with them.
Disclaimer: I have no interest in JEOL
Michael D. Standing e-mail: MDStandi-at-bioag.byu.edu Phone: (801)378-4011
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
PHILADELPHIA SOCIETY FOR MICROSCOPY APRIL 1997 MEETING NOTICE
DATE: Tuesday, April 15, 1997
PLACE: Laboratory for Research on the Structure of Matter (LRSM) Building, 33rd and Walnut Street. Parking is available behind the LRSM Building after 5:00 PM.
TIME: 5:30 PM Social hour, hosted by our meeting sponsor
6:30 PM Dinner (see menu at end of message) Members $12.00 Student members $6.00 Non-members $15.00 RESERVATIONS REQUIRED!!!
7:30 PM Speaker
Chemistry in the Crime Lab Richard Saferstein, Ph.D.
Forensic science in its broadest definition is the application of science to law. As our society has grown more complex, it has become more dependent on rules of law to regulate the activities of its members. Forensic science offers the knowledge and technology of science for the definition and enforcement of such laws. The analytical techniques that can be used in forensic science are numerous and diverse. In general, they must be sensitive enough to cope with minute samples of physical evidence, but they must also be reliable and reproducible to withstand scrutiny by fellow experts in and out of the courtroom. Speed and economy have to be considered too, for the typical forensic scientist must analyze hundreds of cases each year. In this talk a variety of analytical procedures that are applicable to solving forensic science problems will be presented. Included in the discussion will be the application of microscopic analysis to forensic hair and fiber. Also discussed will be the role of visible and infrared microspectrophotometric techniques in forensic problem solving. The speaker will review significant achievements that have been made in utilizing DNA typing for the purposes of linking blood and semen evidence to a single individual. A number of actual case discussions will be included in the talk in order to exemplify the relevancy of forensic science to criminal investigation.
Dinner Menu: Beer, wine, assorted soda and bottled water Munchies
Mini wontons with dipping sauce Chicken stir fry Vegetarian stir fry Fried rice Spinach salad
Cake du jour Coffee, decaf or tea
NEWS FLASH: Reservations can now be sent by e-mail by writing to PSM-RESERVATIONS-at-INAME.COM. Be sure to include your name and affiliation in the body of the message. PSM prefers that you use this convenient method to RSVP.
If you don't have access to e-mail, phone reservations will be taken by Ms. Pat Overend at the University of Pennsylvania, 215/898-8337. Deadline for reservations will be Friday, April 11. If you have any questions regarding the meeting please feel free to contact Rollin Lakis of the Executive Council. Cancellations must be received no later than 5:00 PM, April 14, 1997. RESERVATIONS ARE REQUIRED FOR DINNER. We cannot guarantee you a meal if you do not make a reservation by the deadline listed above.
Message-Id: {1.5.4.32.19970401004330.00680654-at-pop3.unsw.edu.au} X-Sender: s7001031-at-pop3.unsw.edu.au X-Mailer: Windows Eudora Light Version 1.5.4 (32) Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
We have two turbo pumped SEMs - a Leo/Cambridge S-360 (1988) and an Hitachi S-900 (1991). We have never had a problem with either turbo. Local Leo service had a policy of reconditioning the (Pfeiffer) turbos every 3 years for bearing replacement just in case. Their policy now is to leave it until there is a sign of imbalance (audible noise). They are confident now that turbos are no more trouble than any other part so take no special precautionary measures apart from re-oiling the bearing every 6 months. The S-900 has a mag lev. Seiko Turbo. Hitachi advice is that so long as you change the backup batteries every 12 months to ensure full charge (in case of power failure the backup keeps the turbine levitated until it runs down) - the pump will last forever.
Advantage is no fuss clean vacuum. The source of contamination has to be your specimen (or some backstreaming from the rotary, though oils oughtn't get through the turbo if its runnning). AND no problem with water supply/cooling and very little problem with power failure. (Everything coasts to a stop).
BUT a well designed oil diffusion pump system can be NEARLY as good. For that reason I did not insist on the turbo option with our latest Hitachi S-4500. How sure are you of design? Some well known SEMs were plagued with a badly designed diff pump system which stalled and backstreamed with nearly every specimen change!
I am interested in HREM image simulations of III-V semiconductors and I am looking for any references where I can find the values of the Debye-Waller temperature factors for such atoms as P, Ga, IN, As etc. I would be extremely grateful for any helpful information
Sincerely yours
Rafal Spirydon
Dept. of Materials Science and Engineering Kwangju Institute of Science and Technology South Korea
} Jeol sells a high resolution standard (gold on carbon) that we would like } to purchase. Does anyone have info: (part number, price, where to order)? } Many thanks. } } #################################################################### } John J. Bozzola, Ph.D., Director } Center for Electron Microscopy } Neckers Building, Room 146 - B Wing } Southern Illinois University } Carbondale, IL 62901-4402 } 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 } ####################################################################
Most (all) of the EM supplies companies sell these types of specimens - Agar, SPI, Ted Pella, etc. I'd check their prices as well, since you might find them cheaper than EM manufacturers.
More generally, how do others find this specimen performs for resolution checks, particularly at very high resolution and/or low kV? Isn't the carbon a potential/real source of contaminations? I've seen tin spheres on aluminium being promoted as a resolution test specimen without the contamination problem. Is this a good substitute?
Regards,
--------------------------------------------------------------- Dr L. P. Stoter Technical Editor, MICROSCOPY & ANALYSIS Technesis 17, Rocks Park Road email: LPS-at-teknesis.demon.co.uk Uckfield, E. Sussex Phone: +44 (0)1825 766911 TN22 2AT Fax: +44 (0)1825 766911 United Kingdom ---------------------------------------------------------------
I have a turbopump horror story with a happy ending! A number of years ago, we decided to upgrade the diff pump which came on our Etec with a turbo. At that time greased bearing turbos were just appearing on the market. We ordered and installed a 360 l/s pump in place of the diff unit. The Etec e-optics system was quite susceptible to vibration which proved to be a problem, limiting effective maximum mag to between 5-10kX. Worse yet was the reliability. I have forgotten exactly how many (warranty and otherwise) pumps we went through. Must have been more than 5-6 in just a year or two. Bearing failure was the culprit. Some pumps would last for months, others failed only a few hours after installation. { {Greased bearing pumps have since improved, I hear} } Then the maglev pump hit the market! We replaced the ball bearing unit with a Leybold 340 l/s maglev turbo. That was somewhere over eight years ago. The system has been crashed to atm several times (valve failure) and been shut down quite a
number of times. It is still going strong with no problems. ...Love it. Must admit the Etec vacuum valving arrangement allows the pump to run continuously - it is not shut down for sample changes.... As far as the vibration problem, I can't tell I have a turbo pump on the system. The only drawback was the cost of the maglev pump. Given the maintenance free, vibration free, clean vacuum, it was worth it.
Good morning: Once again, I have a bit of a puzzle. I'd like to know _specifically_ the mechanism that generates continuum X-rays or "Bremstralung". There is variation among my text references as to the precise nature of the event that produces this signal. Several texts state that the interaction is inelastic and results from the slowing of electrons as they pass near the nucleus (electron-nucleus interaction), while others state that it is, again, inelastic, and is an primary electron-outer shell electron interaction (specific references are not given to avoid finger-pointing). My understanding of the term "inelastic" is that it refers to electron scattering of primary (beam), backscattered, and secondary electrons, and not to electron-atomic nucleus interaction. If someone could take the time to explain in gory detail, I'd appreciate it very much. I have a list of people to thank and a summary of my Holey grid responses to post, but that will come later. Many thanks for enlightenment. Interactively yours, Dwight
Dwight Beebe E-mail: beebed-at-ere.umontreal.ca Institut de recherche en biologie vegetale Voice: 514-872-4563 Universite de Montreal FAX: 514-872-9406 4101, rue Sherbrooke est Montreal, Quebec H1X 2B2 Canada
At 10:04 AM 4/1/97 -0400, Dwight Beebe wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I don't believe we'll ever know the specific mechanism but the macroscopic model we have is apparently built upon an "apparent" bi-modal distribution of "events" we describe as "elastic" and "inelastic", and the (simple) definitions as I interpret them are: "If energy is transferred, then call it inelastic" ... "If very little energy is tranferred, then call it elastic". Another definition of "inelastic" would claim that characteristic information should be the result of an inelastic event.
(... an example of characteristic info gained would be a characteristic x-ray or a auger electron, whereas a backscattered electron, while informative, is not characteristic and is the result of an elastic event ...)
Since the x-ray contiuum is obviously the result of energy transfer it comes under the inelastic event catagory ... yet while it offers no characteristic information (like BSE, at least macroscopically) it has to be attributed to a "continuum" of energy transfers and while we can't "see" what actually goes on (orbital vs. nuclear), why claim one or the other???
... my $0.02 ...
cheerios, shAf
{\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo ZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
} I am interested in HREM image simulations of III-V semiconductors and I am } looking for any references where I can find the values of the Debye-Waller } temperature factors for such atoms as P, Ga, IN, As etc. } I would be extremely grateful for any helpful information
} Sincerely yours
} Rafal Spirydon
} Dept. of Materials Science and Engineering } Kwangju Institute of Science and Technology } South Korea
A good source of information is 'Debye-Waller Factors of Zinc-Blende-Structure Materials -- A Lattice Dynamical Comparison' by John S. Reid, Acta Cryst. (1983) A 39, 1-13. This contains calculated Debye-Waller Factors for the two atom species for various materials at a range of temperatures.
Another good place to find calculated ELEMENTAL Debye-Waller factors is 'Debye-Waller Factor for Elemental Crystals' by V. F. Sears and S. A. Shelley, Acta Cryst. (1991) A 47, 441-446. Alternatively, Sears and Shelley's results have been tabulated in 'Debye-waller Factors and Absorptive Scattering Factors of Elemental Crystals' by L.-M. Peng, G. Ren, S. L. Dudarev and M. J. Whelan, Acta Cryst. (1996) A 52, 456-470.
My own experience suggests that all of these sources are reasonably accurate (where, for Debye-Waller factors, 'reasonably' probably means less than ~10%-20% error). However, it still leaves you with the problem of deciding exactly what the temperature of your sample is under the electron beam (especially if you've cooled the sample to liquid nitrogen temperatures)!!
Regards,
Martin Saunders, Center for Materials Science and Engineering, Department of Mechanical Engineering, Naval Postgraduate School, Monterey, CA 93943, USA.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Colleagues: Is anyone familiar with Sequenza staining racks? These are used to hold microscope slides for staining, and I'm not sure how they are different from your basic, run-of-the-mill slide racks. I haven't been able to find them in any catalogs.
TIA, Bev Maleeff
SmithKline Beecham Pharmaceuticals Toxicology-US, UE 0462 709 Swedeland Road King of Prussia, PA 19406 610/270-7987 610/270-7202 fax Beverly_E_Maleeff-at-sbphrd.com
} Good morning: } Once again, I have a bit of a puzzle. I'd like to know } _specifically_ the mechanism that generates continuum X-rays or } "Bremstralung". There is variation among my text references as to the } precise nature of the event that produces this signal. Several texts state } that the interaction is inelastic and results from the slowing of electrons } as they pass near the nucleus (electron-nucleus interaction), while others } state that it is, again, inelastic, and is an primary electron-outer shell } electron interaction (specific references are not given to avoid } finger-pointing). My understanding of the term "inelastic" is that it } refers to electron scattering of primary (beam), backscattered, and } secondary electrons, and not to electron-atomic nucleus interaction. If } someone could take the time to explain in gory detail, I'd appreciate it } very much. } I have a list of people to thank and a summary of my Holey grid } responses to post, but that will come later. Many thanks for } enlightenment. } Interactively yours, } Dwight } } } Dwight Beebe E-mail: } beebed-at-ere.umontreal.ca } Institut de recherche en biologie vegetale Voice: 514-872-4563 } Universite de Montreal FAX: 514-872-9406 } 4101, rue Sherbrooke est } Montreal, Quebec H1X 2B2 } Canada
This is going back a few years, but .....
The "Bremstralung" originates ONLY because the electrons are accelerated (or deaccelerated). The mechanism that causes the change in velocity is irrelevant. The cause comes from Special Relativity, as applied to charged particles - change the velocity of a charged particle and it will radiate.
The wavelength(?) and intensity(?) of the radiation will depend on factors like the change in velocity, mass and velocity of particle. The effect is only significant at near-relativistic velocities. Note also, it is specifically a change in VELOCITY, a vector quantity, not speed. You see a similar effect in sychrotrons, where the speed of the charge particle is constant but its direction is changed, so resulting in sychrotron radiation.
Additionally, don't mistake the background you see from an EDX/WDX detector on an electron microscope. Some of its characteristics are related to Bremstralung, but, especially at the low energy end, the overall response of the detector and amplifier are much more significant.
Regards, Larry Stoter
--------------------------------------------------------------- Dr L. P. Stoter Technical Editor, MICROSCOPY & ANALYSIS Technesis 17, Rocks Park Road email: LPS-at-teknesis.demon.co.uk Uckfield, E. Sussex Phone: +44 (0)1825 766911 TN22 2AT Fax: +44 (0)1825 766911 United Kingdom ---------------------------------------------------------------
MSA Recipients; I have a LEKTRA "Densi-timer" model PTM-4A which is in need of repair or replacement, but the company is no longer in business. The Densi-timer is a three-piece light meter for black and white only photo enlarging which reads an exposed area on the easel/paper to determine exposure time based on paper characteristics, rheostat setting, and magic. Any assistance in repair or suggestions for replacement will be greatly appreciated. Thanks. Merci. Shukran. Gracias. Danke. ------------------------------------- Name: Winston Wiggins Carolinas HealthCare System Charlotte, NC USA E-mail: wwiggins-at-carolinas.org Fax: 704-355-7648 Voice:704-355-7220
Message-Id: {199704012200.RAA27986-at-umic.sunysb.edu} Comments: Authenticated sender is {greg-at-mail.umic.sunysb.edu}
Hi all, I am looking for a special centrifuge tube that allows you to place a TEM grid in it. When the tube is spun the sample settles on the grid. Does anyone know who carries this tube.--Thanks in advance Gregory Rudomen Greg-at-UMIC.SUNYSB.EDU 516-444-3126 University Microscopy Imaging Center S.U.N.Y. Stony Brook
} Once again, I have a bit of a puzzle. I'd like to know } _specifically_ the mechanism that generates continuum X-rays or } "Bremstralung". There is variation among my text references as to the } precise nature of the event that produces this signal. Several texts state } that the interaction is inelastic and results from the slowing of electrons } as they pass near the nucleus (electron-nucleus interaction), while others } state that it is, again, inelastic, and is an primary electron-outer shell } electron interaction (specific references are not given to avoid } finger-pointing). My understanding of the term "inelastic" is that it } refers to electron scattering of primary (beam), backscattered, and } secondary electrons, and not to electron-atomic nucleus interaction. If } someone could take the time to explain in gory detail, I'd appreciate it } very much.
Dear Dwight, The simple explanation of bremsstrahlung is that accelerated charge produces electromagnetic radiation. The mechanism is that when an electron encounters an electromagnetic field, it is accelerated, thus it can radiate. There is no difference whether the field is due to other electrons, nuclei or "wiggler" magnets. Most bremsstrahlung produced by the interaction of electrons with matter is produced by the interaction with nuclei. The field near a nucleus is greater than that near an elec- tron by a factor of Z. Furthermore, from considerations of momentum and energy conservation, there is a higher probability of the reaction
e + M -} e + M + photon
if M is a large mass. Synchrotron light sources use "wiggler" magnets-- an arrangement of magnets which produce fields directed alternately up and down--to direct fast electrons on a sinuous path with a particular frequency. The electrons are accelerated and radiate photons which are more monochromatic than those produced by interactions with nuclei. As was pointed out, any scattering which results in the kinetic energies of the initial particles being greater than those of the same particles in the final state is inelastic. Photon production, changes in internal energy of the target nuclei and nuclear reactions are all examples of inelastic scattering. Yours, Bill Tivol
Very simply, elastic interactions refer to those where energy is not lost (i.e., transformed) during the process. Inelastic interactions involve a "loss" or transformation of energy. Since a portion of the energy of the electron is lost/converted into a bremstrallung photon, the event is by definition inelastic. Now as to where that interation takes place, I will let someone else comment. I thought it was by close encounters with the nucleus.
At 10:04 AM 4/1/97 -0400, you wrote: My understanding of the term "inelastic" is that it } refers to electron scattering of primary (beam), backscattered, and } secondary electrons, and not to electron-atomic nucleus interaction. If } someone could take the time to explain in gory detail, I'd appreciate it } very much. ---------------------------------------------------- Warren E. Straszheim 270 Metals Development, Ames Lab/ISU, Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-3091
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
RE} } Continuum X-ray generation? 4/1/97
Whenever a charged particle is accelerated, it emits radiation. Bremsstrahlung is the term used for the radiation emitted during atomic collisions, where a charged particle is accelerated by the charges it "sees" in its approach. You can find more than you would ever care to know in Jackson's "Classical Electrodynamics," although a more readable (and actually enjoyable) version of the basic physics (quantum electrodynamics) appears in Richard Feynman's "QED."
--------------------------------------
At 10:04 AM 4/1/97 -0400, Dwight Beebe wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I don't believe we'll ever know the specific mechanism but the macroscopic model we have is apparently built upon an "apparent" bi-modal distribution of "events" we describe as "elastic" and "inelastic", and the (simple) definitions as I interpret them are: "If energy is transferred, then call it inelastic" ... "If very little energy is tranferred, then call it elastic". Another definition of "inelastic" would claim that characteristic information should be the result of an inelastic event.
(... an example of characteristic info gained would be a characteristic x-ray or a auger electron, whereas a backscattered electron, while informative, is not characteristic and is the result of an elastic event ...)
Since the x-ray contiuum is obviously the result of energy transfer it comes under the inelastic event catagory ... yet while it offers no characteristic information (like BSE, at least macroscopically) it has to be attributed to a "continuum" of energy transfers and while we can't "see" what actually goes on (orbital vs. nuclear), why claim one or the other???
... my $0.02 ...
cheerios, shAf
{\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo ZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
------------------ RFC822 Header Follows ------------------ Received: by qmgate_backup.anl.gov with ADMIN;1 Apr 1997 16:11:23 U Received: by cmtnts3.cmt.anl.gov with SMTP (Microsoft Exchange Server Internet Mail Connector Version 4.0.994.63) id {01BC3EB7.6F911E50-at-cmtnts3.cmt.anl.gov} ; Tue, 1 Apr 1997 16:12:03 -0600 Received: from sparc5.microscopy.com by cmtnts3.cmt.anl.gov with SMTP (Microsoft Exchange Internet Mail Connector Version 4.0.994.63) id HPXZPMMF; Tue, 1 Apr 1997 16:11:59 -0600 Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id MAA16270 for dist-Microscopy; Tue, 1 Apr 1997 12:45:34 -0600 Received: from darkwing.uoregon.edu (darkwing.uoregon.edu [128.223.142.13]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id MAA16267 for {Microscopy-at-Sparc5.Microscopy.Com} ; Tue, 1 Apr 1997 12:45:30 -0600 Received: from shaf.uoregon.edu (shaf.uoregon.edu [128.223.94.20]) by darkwing.uoregon.edu (8.8.5/8.8.5) with SMTP id KAA00370 for {Microscopy-at-Sparc5.Microscopy.Com} ; Tue, 1 Apr 1997 10:56:44 -0800 (PST) Message-ID: {c=US%a=_%p=Argonne_National%l=CMTNTS39704012212HPXZPMMF-at-cmtnts3.cmt.anl.gov}
There are very good discussions of "Continum X-ray Production" on p. 117 of the Book 'Scanning Electron Microscopy and X-ray Micro Analysis' by Goldsterin, et. al. 2nd Ed, Plenum Press, 1992 (a copy of which should be in every SEM & EMPA lab); and on p. 157-161 of the book 'Electron Beam X-ray Microanalysis' bu Kurt Heinrich, Van Nostrand Reinhold, 1981. Both sources imply that Bremsstrahlung photons are produce by the deacceleration of beam electrons by an interaction with the nuclear field of the atom. In the classical sense such interactions would be considered to be inelastic since they involve a measureble change in energy of the incident electron.
I believe that backscattered electrons are usually considered to be produced by inelastic interactions between the beam electrons and the positive charge of the atoms' core (i.e. the nuclear charge moderated by the tightly-bound inner-shell electrons). This process is described in some detail, both from the classical and the quantum mechanical point of view, by Reimer in his book 'Scanning Electron Microscopy', Springer Verlag, 1985, p. 57-73. Reimer is a pretty well grounded physicist, and so I would think his opinion would be reliable. Reimer treats inelastic scattering processes in similar detail on pages 73-81. The question of the relative magnitudes of the energy transfer involved in elastic and inelastic scattering processes is also discussed on p. 73.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
Rafal Spirydon wrote: } } I am interested in HREM image simulations of III-V semiconductors and I am } looking for any references where I can find the values of the Debye-Waller } temperature factors for such atoms as P, Ga, In, As etc.
Have a look at:
JS Reid, "Debye-Waller Factors of Zinc-Blende-Structure Materials - A Lattice Dynamical Comparison", Acta Cryst A 39 (1983) 1-13.
This reference has Debye-Waller factors for seventeen materials over the temperature range 1 to 1000 K (where appropriate), including GaP, GaAs, GaSb, InP, InAs, and InSb.
Stephen. ............................................................... : Stephen Anderson Australian Key Centre for : : Microscopy and Microanalysis : : Email stephen-at-emu.usyd.edu.au The University of Sydney : : Telephone (+61)-2-9351 7552 NSW 2006 : : Facsimile (+61)-2-9351 7682 Australia : :.............................................................:
We are examining the bacteria Pseudomonas fluorescens (gram-negative soil bacteria) with an SEM. We usually fix with glut, adhere the bacteria to coverslips, dehydrate, CPD, Au/PD coat, and examine. In this case, however, we are having problems with charging and focusing. I haven't had this problem with other bacterial samples. I am going to try osmicating the bacteria but does anyone have any other suggestions?
Thank you in advance,
Ginger Baker EM Lab Manager Dept. APP 250 Vet Med Oklahoma State University Stillwater, OK 74078 (405) 744-6765 FAX: (405) 744-5275 Email: lizard-at-okway.okstate.edu
Rafal Spirydon wrote: } } I am interested in HREM image simulations of III-V semiconductors and I am } looking for any references where I can find the values of the Debye-Waller } temperature factors for such atoms as P, Ga, In, As etc.
Have a look at:
JS Reid, "Debye-Waller Factors of Zinc-Blende-Structure Materials - A Lattice Dynamical Comparison", Acta Cryst A 39 (1983) 1-13.
This reference has Debye-Waller factors for seventeen materials over the temperature range 1 to 1000 K (where appropriate), including GaP, GaAs, GaSb, InP, InAs, and InSb.
Stephen. ............................................................... : Stephen Anderson Australian Key Centre for : : Microscopy and Microanalysis : : Email stephen-at-emu.usyd.edu.au The University of Sydney : : Telephone (+61)-2-9351 7552 NSW 2006 : : Facsimile (+61)-2-9351 7682 Australia : :.............................................................:
The site features a discussion area, links to ESEM sites and images on the net, meeting information, microscopy job list, and an archive of all ESEM related posts to the microcopy listserver. Check it out and send me any feedback you might have.
Scott
------------------ Scott A. Wight email: swight-at-erols.com Homepage: http://www.geocities.com/CapeCanaveral/3429
Slovenian Society for Electron Microscopy Austrian Society for Electron Microscopy Croatian Society for Electron Microscopy Czechoslovak Society for Electron Microscopy Microscopy Society of Hungaria Italian Society for Electron Microscopy
TIME SCHEDULE
May, 30 1997: Submission of Papers Registration
June 30, 1997: Notification of acceptance
August 30, 1997: Payment Accommodation reservation
September 15, 1997: Preliminary Program
October 5-8, 1997: Congress
For MORE INFORMATION, such as
List of invited speakers List of Exhibitors and Sponsors Instruction for preparation of papers Congress fee and Hotel prices Registration and Accommodation forms Ground plan of the Congress Centre and much more
please visit Congress Home Page: http://www2.ijs.si/~k5www/MCEM97/index.html
or send us an email and we will forward you complete information.
ADDRESS OF THE CONGRESS SECRETARIAT:
MCEM'97 Ceramics Department "Jozef Stefan" Institute Jamova 39, 1000 Ljubljana Slovenia
Most likely the gold coating is sufficient on top of the bacteria but the bacteria are little "umbrellas" which prevent the gold from forming a continuos coating, connecting the under side of the bacteria and the substrate. It helps if the specimen is osmicated. Infusion of silver nitrate has been used to make the specimen more conductive. The carbon tabs or carbon coated mica solve part of the problem because the substrate is conductive. With an evaporator one could rotate the specimen and evaporate gold from two sources. One source should be at a very shallow 6-10 degrees to the specimen. When sputter coating try placing the specimen at about 45 degrees, give it a somewhat lighter coating and then lift the other side and apply a second coating. Using this method a better coating can form under the specimen. Sometimes people forget to paint a conducting path when the coverslip overhangs the specimen stub, but that is not so much a technical problem, rather it's a self-inflicted wound. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 300+ Links, MSDS ************************ http://www.proscitech.com.au
} } We are examining the bacteria Pseudomonas fluorescens } (gram-negative soil bacteria) with an SEM. We usually fix with glut, } adhere the bacteria to coverslips, dehydrate, CPD, Au/PD coat, and } examine. In this case, however, we are having problems with charging } and focusing. I haven't had this problem with other bacterial samples. } I am going to try osmicating the bacteria but does anyone have any } other suggestions? } } Thank you in advance, } } Ginger Baker } EM Lab Manager } Dept. APP } 250 Vet Med } Oklahoma State University } Stillwater, OK 74078 } (405) 744-6765 } FAX: (405) 744-5275 } Email: lizard-at-okway.okstate.edu }
It is probably your substrate. Try putting them on a 0.2=B5m nucleopore filter. Background is not as pretty but charging is not as much of a= problem. Another alternative (standard practice around here) is to paint a continuous line of conductive carbon or silver paint from the top of the coverslip around the edge to the stub to provide a better grounding path. Do this in a couple of obscure spots to your existing samples and see if it works.=20 You may have to adhere the bact. to the coverslips better as well. Go to the web address at the end of this message and find the "Tips & Tricks link. In the TEM section are a bunch of links called "Stickey" something or other which may be useful. Good luck
At 04:43 PM 4/1/97 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 352-392-1295 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
It is probably your substrate. Try putting them on a 0.2=B5m nucleopore filter. Background is not as pretty but charging is not as much of a= problem. Another alternative (standard practice around here) is to paint a continuous line of conductive carbon or silver paint from the top of the coverslip around the edge to the stub to provide a better grounding path. Do this in a couple of obscure spots to your existing samples and see if it works.=20 You may have to adhere the bact. to the coverslips better as well. Go to the web address at the end of this message and find the "Tips & Tricks link. In the TEM section are a bunch of links called "Stickey" something or other which may be useful. Good luck
At 04:43 PM 4/1/97 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 352-392-1295 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
My manager wishes to understand my work. Is there anyone who offers a short course in the fundamentals and basics of TEM analysis for materials applications. Any help will be greatly appreciated.
Thanks, Kim Christensen
Kim Christensen White Oak Semiconductor 600 East Main Street, Suite 800 Richmond, VA 23219 Ph: 804-698-7307 Fax: 804-698-7316
We have two devices for sedimenting material onto a TEM grid. The first is a Beckman Airfuge with the EM90 rotor. The grid is placed on a 5 mm square of 0.025 um nitrocellulose and covered with a film of parlodian. This sandwich is placed in the rotor w/ the grid facing the sample prior to sedimentation.
The second device makes use of the 3 mm tubes for a Beckman ultracentrifuge. The tubes have round bottoms but we make semi- spherical inserts out of epoxy that fit in the bottom and leave a flat surface for the grid to rest on. The inserts are made by pouring a drop of epoxy in a tube, allowing it to polymerize and cutting it out of the tube. We then sand them smooth to reduce their size slightly so they will easily slide into another tube. The inserts are reusable and sterilizable.
Regards, Joe Neilly Microscopy and Microanalysis Abbott Laboratories North Chicago, IL 60064
Does anyone know of a selective etch that will etch away silicon but not silicon nitride? I'm looking at a multilayer sample in plan view for TEM and hope to back etch the si substrate and use the nitride as a stop layer.
Buy your manager the books "Transmission Electron Microscopy", by David B. Williams and C. Barry Carter, Plenum Press, NY. ISBN 0-306-45324-X. And upon receiving it you will want another copy for yourself as well.
On Wed, 2 Apr 1997, Christensen, Kim wrote:
} Dear colleagues, } } My manager wishes to understand my work. Is there anyone who offers a short } course in the fundamentals and basics of TEM analysis for materials } applications. Any help will be greatly appreciated. } } Thanks, } Kim Christensen } Yves MANIETTE Universitat de Barcelona Serveis Cientifico Tecnics Unitat ESCA TEM Carrer Lluis Sole i Sabaris, 1-3 E-08028 BARCELONA ESPANYA
Centralized microscope facilities have become increasingly concerned with cost recovery. Most of us that oversee such facilities are aware that Federal regulations require equal charges for in-house users. This was the subject of a number of recent inquires addressed to this newsgroup. My question is how classes are handled. Do you charge the University department or unit whose class uses the facility? If so, how do you figure the charge? Please reply to this newsgroup or to my e-mail or FAX.
Thanking you in advance for your time
Lewis Coons, Ph.D., Director Integrated Microscopy Center Life Sciences Bldg. Campus Box 526040 University of Memphis Memphis TN 38152-6040 FAX 901 678 4457j
} We are examining the bacteria Pseudomonas fluorescens } (gram-negative soil bacteria) with an SEM. We usually fix with glut, } adhere the bacteria to coverslips, dehydrate, CPD, Au/PD coat, and } examine. In this case, however, we are having problems with charging } and focusing. I haven't had this problem with other bacterial samples. } I am going to try osmicating the bacteria but does anyone have any } other suggestions? } } Thank you in advance, } } Ginger Baker
Ginger, Two suggestions that I've found helpful, both having to do with the non-conductuve nature of glass coverslips: 1) Connect the top surface of the coverslip to the stub with silver paint and cover as much of the top surface of the coverslip as you can sacrifice with silver paint. Leave only the area(s) bare that need to be left uncovered to examine the bacteria. 2) Don't use glass coverslips. Coat both sides of membrane filters in the sputtercoater (careful venting! they like to fly around), mount the coated filters on stubs using carbon-conductive double-sticky discs (Pella, and maybe others). Dry the bacteria from air, alcohol, or HMDS; if from fluid, the final change in drying fluid with bacteria in suspension is dropped directly onto the filter pieces on the stubs, then allowed to dry. Be sure to use filters that have nice holes punched in them (Nucleopore, Poretics, that kind), *not* torturous-path filters like, say, Millipore, or you'll have a hard time telling the difference between bacteria and filter. After drying, sputter coat as usual. Phil
&&& Illigitimi non carborundum &&&&&&&& Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217) 355-1143 oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
Well folks, I have just finished the cost analysis for the EM service lab as well as 13 other services labs that are part of our Biotechnology Center. I am not sure it would stand up to a federal audit, but I generated enough numbers to keep the local auditors and accountants baffled for quite some time.
Essentially we put together all of the cost incurred in running each lab, including building services, operations and maintenance, equipment depreciation, salaries and fringe benefits, and a portion of the administartive costs of our center office. Our grants office was able to come up with those numbers for each of the rooms we occupy. After I had a total figure for each lab I deducted a certain percentage based on the activities in each lab NOT devoted to providing services for which we recover costs. That would include teaching, methods development, student advising etc. We have a generous suplement to our budget from the university so we make no charge for those activities. It was then this adjusted cost of operation upon which I based "true cost" of each service. This cost was then used for a maximum rate determination that we would charge "in house" users. All of our "in house " charges are well below the "true cost" of the service, so I expect we would not have any trouble defending the charges we make to federal grantees, should the feds ever show up at our door. ANyone interested in how I cost out each service should contact me since it is rather complicated and lengthy for this forum.
Good luck to all of you who have to go thru this exercise. } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } At 09:43 AM 4/2/97 -0600, you wrote:
} Dear Colleagues: } } Centralized microscope facilities have become increasingly concerned with } cost recovery. Most of us that oversee such facilities are aware that } Federal regulations require equal charges for in-house users. This was the } subject of a number of recent inquires addressed to this newsgroup. My } question is how classes are handled. Do you charge the University } department or unit whose class uses the facility? If so, how do you figure } the charge? Please reply to this newsgroup or to my e-mail or FAX. } } Thanking you in advance for your time } } } Lewis Coons, Ph.D., Director } Integrated Microscopy Center } Life Sciences Bldg. } Campus Box 526040 } University of Memphis } Memphis TN 38152-6040 } FAX 901 678 4457j } } } } ******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
Hello, I am a student of the engineering school of material EEIGM of Nancy (france) realizing at the present time an engineering training period in the material laboratory of the UPC (University of Barcelona-spain). I am working on asbestos characterization by TEM but I have some problems to obtain good sample preparation using the "METHOD 7402-NIOSH":
Sample : Filter with a cellulose ester membrane fibers of asbestos
Sample preparation : 1-remove a section of the filter 2-affix the filter section to a clean glass slide 3-place the slide in a petri dish wish contains several paper filters soaked with 2 to 3 ml acetone. Cover the dish and wait 2 to 4 min for the sample filter to fuse and clear. 4-transfer the slide to a rotating stage inside the bell jar ofa vacuum evaporator. Evaporate a 1 by 5 mm section of a graphite rod 5-place the filter, carbon side down, on a grid (200mesh) in a acetone satured petri dish during hours to disolve the filtre.
the problem is that the carbon film is torn on the TEM grid. Could you help me to resolve this problem??
Thank you in advance
Laurent STEINMETZ ----------------------------------------------------------------------- | Jose M Manero E-mail: manero-at-cmem.upc.es | | Electronic Microscopy Lab | | Department of Materials Science and Metallurgical Engineering, UPC | -----------------------------------------------------------------------
RE: } } We are examining the bacteria Pseudomonas fluorescens } (gram-negative soil bacteria) with an SEM. We usually fix with glut, } adhere the bacteria to coverslips, dehydrate, CPD, Au/PD coat, and } examine. In this case, however, we are having problems with charging } and focusing. I haven't had this problem with other bacterial samples. } I am going to try osmicating the bacteria but does anyone have any } other suggestions? } } Thank you in advance, } } Ginger Baker } EM Lab Manager } Dept. APP } 250 Vet Med } Oklahoma State University } Stillwater, OK 74078 } (405) 744-6765 } FAX: (405) 744-5275 } Email: lizard-at-okway.okstate.edu
I have found it very useful to "pre-coat" either cover slips or Nuclepore polycarbonate filters with either Au or Au/Pd prior to applying samples. This provides a conducting substrate beneath the sample. In the case of Nuclepore filters I coat both sides with ~15-30 nm of metal in a sputter coater unit. With coverslips I always "paint" a thin band of colloidal Ag connecting the upper surface to the underlying SEM stub.
This technique also works for Low Voltage cryo SEM on uncoated specimens. I mix ~1:10 ratio of colloidal Ag with a cryoglue (TBS, OCT, Tissue Tek...) and store this mixture in a 1ml tuberculin syringe without a needle. I apply a drop or two of this, spread it around and use it to affix pieces of my precoated polycarbonate membranes with fresh cells to a Si chip just prior to plunge freezing a sample.
Additionally, the use of Si chips (Ted Pella) instead of cover slips provides a nice smooth conducting surface that cultured cells can be grown on.
Ed Basgall, PhD Penn State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618
The Appalachian REgional Microscopy Society announces the
AREMS SPRING 1997 MEETING, Thursday & Friday, April 17 & 18
For registration and information contact: Susan Read, BASF Corporation, Sand Hill Road, Enca, NC 28728 Telephone: (704) 667-6353 Fax: (704) 667-6903 E-mail: reads-at-basf.com
MEETING EVENTS AND PROGRAM FOR THURSDAY 17 APRIL:
Noon to 5:00 p.m.: Registration, Comfort Suites, NC 191 at Biltmore Square Mall
1:00 p.m. to 5:00 p.m.: Workshops at Comfort Suites and BASF
Workshop 1: Tours of the BASF Corp. Fiber Products Research & Development Microscopy Laboratories (each tour limited to 15 participants)
Workshop 2: Scanning Electron Microscopy: Philips XL30/EDAX DX4 Integrated System - Philips Electronic Instruments, SEM Laboratory at BASF Fiber Products R & D (limit: 10 participants; If there is enough interest, another session may be added.)
Workshop 3: Basic PC Image Capture in Light Microscopy - Martin Microscope Company, Comfort Suites
Workshop 4: Internet Workshop - Mike Webber of LEO Electron Microscopy, Comfort Suites
6:00 p.m. to 7:00 p.m.: AREMS Social Hour, Enka Lake Club, the Patio (Weather Permitting), Wine tasting: wines from the Biltmore Estate Winery, Asheville, North Carolina Beer Tasting: beers from Highland Brewery, Asheville, North Carolina Light hors d'oeuvres
7:00 p.m. to 9:00 p.m.: AREMS Banquet & Keynote Address, Enka Lake Club, Buffet dinner of prime rib, grilled chicken, vegetables, tossed salad, fruit, dessert, and beverage. Dinner Music by Harpist Carroll Owenby
Early 19th Century Country Fashions: History and Evaluation of Mast Suit - a joint address by: Kathleen Wilson, Research Associate, East Tennessee State University, and curator of the Kings Mountain Cultural Center, Johnson City, Tennessee Patricia Ewer, Textile Conservator, The Biltmore Estate, Asheville, North Carolina Susan Read, Technologist, BASF Corporation, Fiber Products Research & Development, Enka, North Carolina
MEETING EVENTS AND PROGRAM FOR FRIDAY 18 APRIL:
8:00 a.m. to 9:30 a.m.: Registration & Coffee, Enka Lake Club (coffee, juice, fruit & danish will be served)
8:30 a.m. to 9:00 a.m.: Overview of BASF Fiber Products Research and Development - Otto Ilg, Vice-President Technology, BASF Corporation, Fiber Products Division
9:00 a.m. to 9:30 a.m.: The Funny and the Grim Aspects of Microscopy - Don Felty, MicroSolutions
9:30 a.m. to 10:00 a.m.: Examining DNA with a Fullerene Imaging Agent - Alan Cassell, Graduate Research Assistant, Department of Chemistry and Biochemistry, University of South Carolina
10:00 a.m. to 10:30 a.m.: Security and Antiforgery Features in Currency and Security Documents - Dr. Matt Hoyt, Senior Research Chemist, BASF Corporation, Fiber Products R&D
10:30 a.m. to 11:00 a.m.: Break for Visiting with Exhibitors
11:00 a.m. to 11:45 a.m.: AREMS Business Meeting
11:45 a.m. to 12:15 p.m.: Design and Implementation of a Laboratory-Wide Image Management System Using Microscoft Windows NT and Magneto-Optical Storage Technology - Rick McGill, Microscopy and Morphology Research, Eastman Chemical Company, Kingsport, Tennessee
12:15 p.m. to 1:00 p.m.: Snails, Eggs, and Microscopy: Light, SEM, and TEM - Stan C. Kunigelis, Associate Professor of Zoology, Clinch Valley College of the University of Virginia, Wise, Virginia
1:00 p.m.: Closing Remarks & Lunch, Enka Lake Club (Assorted salads and sandwich fixings)
Remainder of afternoon and evening free for visiting Asheville. Visits to the Biltmore Estate are recommended
Hello, We are attempting to immunolabel a ribo- nucleocapsid. We have been successful employing negative staining. This is a single-stranded RNA molecule complexed to many copies of the same protein. We have an antibody that stains westerns very well at 1:10000. Should we try incubating this complex with the antibody at say 1:100 and then apply to a grid or after. I guess it would be easy to try both, but if anyone has experience with this we would appreciate any suggestions. Cheers, Hank Adams Electron Microscopy Laboratory New Mexico State University Las Cruces, NM 88003
Good suggestion! I bought one myself and it is an excellent resource. I also understand that Ron Anderson will be editing a companion to that book which will be a comprehensive look at TEM Specimen Preparation. I think that will be a "must buy" also.
David Henriks TEL: 800-728-2233 (toll-free in USA) South Bay Technology, Inc. 714-492-2600 1120 Via Callejon FAX: 714-492-1499 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 Yves Maniette } Kim,
Buy your manager the books "Transmission Electron Microscopy", by David B. Williams and C. Barry Carter, Plenum Press, NY. ISBN 0-306-45324-X. And upon receiving it you will want another copy for yourself as well. {
Hi all, I am looking for a special centrifuge tube that allows you to place a TEM grid in it. When the tube is spun the sample settles on the grid. Does anyone know who carries this tube.--Thanks in advance Gregory Rudomen Greg-at-UMIC.SUNYSB.EDU 516-444-3126 University Microscopy Imaging Center S.U.N.Y. Stony Brook
The only one that I am familar with is the Aifuge centrifuge made by Beckman. This airfuge is designed especially for small volume samples and they have a really nicely designed rotor just for putting a grid in the bottom and pelleting your sample right onto your grid
If you already have the airfuge, the rotor is part #347844 and the price was $2,270.00 (back in 1995). I am not quite sure of the price of the airfuge itself, but be warned, Beckman is not known for being reasonable. I think that the price was somewhere in the neighborhood of $20K.
Good Luck, Peggy Bisher
Margaret E. Bisher
NEC Research Institute 4 Independence Way Princeton, NJ 08540. Tel.: (609) 951-2629 Fax: (609) 951-2496 e-mail: peggy-at-research.nj.nec.com
Dear Laurent, When I have done this, I put the filter square on a carbon-film-coated TEM grid filter side UP. The filter gradually dissolves and lowers the carbon film onto the coated TEM grid. The asbestos fibers are caught between the two carbon films. Also, if you use a polycarbonate Nucleopore-type filter, you can skip the "clearing" step and carbon-coat the filter material directly. However, with these, I find you must dissolve the filter in chloroform for 48 hours. You wrote:
} Hello, } I am a student of the engineering school of material EEIGM of Nancy (france) realizing at the present time an engineering training period in the material laboratory of the UPC (University of Barcelona-spain). } I am working on asbestos characterization by TEM but I have some problems to obtain good sample preparation using the "METHOD 7402-NIOSH": } } Sample : Filter with a cellulose ester membrane } fibers of asbestos } } Sample preparation : } 1-remove a section of the filter } 2-affix the filter section to a clean glass slide } 3-place the slide in a petri dish wish contains several paper filters soaked with 2 to 3 ml acetone. Cover the dish and wait 2 to 4 min for the sample filter to fuse and clear. } 4-transfer the slide to a rotating stage inside the bell jar ofa vacuum evaporator. Evaporate a 1 by 5 mm section of a graphite rod } 5-place the filter, carbon side down, on a grid (200mesh) in a acetone satured petri dish during hours to disolve the filtre. } } } } } the problem is that the carbon film is torn on the TEM grid. } Could you help me to resolve this problem?? } } Thank you in advance } } Laurent STEINMETZ Regards, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
H. ADAMS wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello, We are attempting to immunolabel a ribo- } nucleocapsid. We have been successful employing negative staining. This is } a single-stranded RNA molecule complexed to many copies of the same } protein. We have an antibody that stains westerns very well at 1:10000. } Should we try incubating this complex with the antibody at say 1:100 and } then apply to a grid or after. I guess it would be easy to try both, but } if anyone has experience with this we would appreciate any suggestions. } Cheers, } Hank Adams } Electron Microscopy Laboratory } New Mexico State University } Las Cruces, NM 88003 } } http://www.nmsu.edu/Research/artsci/public_html/eml/ } } 505-6463600
Hi Hank,
If I understand correctly you want to label the nucleoprotein on your RNP complex. When I label the measles nucleoprotein on purified RNPs I do it like this : adsorb the RNPs onto a formvar carbon coated grid, remove the excess and immediately add the first antibody (anti protein) generally diluted 1/100 or more (I do 3 dilutions in fact). wash add the second antibody (coupled to gold) wash negative stain as usual
Normally if the first antibody works well, you don't need the second one, at the EM level you can see the RNP much bigger because of the IgG fixed on it. (after staining of course)
Good Luck Daniele SPEHNER Electron Microscopy Laboratory Etablissement de Transfusion Sanguine 67065 Strasbourg Cedex - FRANCE
Wyeth-Ayerst Research, a major division of Fortune 100 American Home Products Corporation, has an opportunity at our pharmaceutical research facility in Chazy, New York for a Scientist in our Imaging Laboratory.
The selected candidate will be responsible for operating a transmission electron microscope (TEM), scanning electron microscope (SEM), and processing images. Resposibilities include processing, embedding, and sectioning tissues in support of electron microscopy services, as well as performing histological techniques. The incumbent must keep records in compliance with SOP's and GLP's and maintain laboratory instruments in accordance with SOP's, troubleshooting equipment problems as needed.
Requirements include a B.S./M.S. with 2-7 years relevant experience and familiarity with all techniques necessary for the operation of TEM, including tissue preparation and image processing. SEM experience is desireable.
The Wyeth-Ayerst Drug Safety Division is located between the foothills of the Adirondack Mountains and the shores of Lake Champlain. The area offers a wide range of outdoor recreational activities while being a short distance from Lake Placid, NY; Burlington, VT; Montreal, Quebec, and Plattsburgh, NY. Wyeth-Ayerst Research offers an excellent compensation and benefits package in a highly professional environment. Please send your resume with salary requirements to:
Mr. Lou Ballester Human Resources Wyeth-Ayerst Research 641 Ridge Road Chazy, NY 12921
You wrote: } } Hi all, } I am looking for a special centrifuge tube that allows you } to place a TEM grid in it. When the tube is spun the ......
Hi Greg (and all),
In the last Fullam catalog I have (1992-93), EFFA Centrifuge Tubes are pictured on page 49. Cost of the tubes then was $135.00/balanced pair (#11450). They also make some to hold No. 00 BEEM capsules, which are the ones I've used in the past (pretty nifty). These are good up to 6000g. They also listed some others that are good up to 34,000g with which I've had no experience. Don't know if they're still available, you'll have to check.
Heather Owen
p.s. I have no connection with Ernest F. Fullam, Inc. - just a satisfied customer.}
Heather A. Owen, Director Electron Microscope Laboratory Department of Biological Sciences University of Wisconsin - Milwaukee (414)229-6816
To all of you who have asked for details on our cost accounting:
More of you responded that I thought might so I have posted the data at our WWW site. I didn't know how to email spreadsheets.
Some places my columns are a little screwed up but I think you can figure it out.
so go to http://www.biotech.ufl.edu/~emcl/cost.html
Let me know if you do not have access to the WWW and I will fax. ******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
Dear Microscopy Folks: I am Candace Haigler, Director of the EM Lab at Texas Tech University, writing this message together with Mark Grimson, EM Technician, who is a member of your Internet group. We find ourselves facing a potential problem about which we need your advice.
Our EM lab is now fairly stable, being purpose-built underground as an attachment to the main Biology building (it sticks out beyond the above-ground footprint of the building). Our only current problem is that our roof is a brick patio that attracts skate-boarders. When they are skate-boarding, we cannot take pictures due to vibrations, but this is a sporadic problem and we can chase them away.
Now we find that the university is making a Master Plan, the draft of which shows a major service road to a new 1000 space parking deck coming very close to our building. We estimate that the road will be within 50 feet, or even closer, to the below-ground EM lab. Given our existing problem with skate boarders, we are very worried that this road will essentially destroy the utility of the lab.
We solicit your help in: (1) Sharing knowledge about similar situations (2) Pointing us to the best published sources about EM lab design, particularly in regard to vibration and preferred distance from nearby roads (3) Pointing us to any expert EM lab design firms from whom we might get information
We are very concerned about this situation, and will greatly appreciate your help. You may reply to Mark at the address shown above, or my personal e-mail address is brchh-at-ttu.edu.
Sincerely, Candace Haigler Professor and Director of the Electron Microscopy Laboratory
We have recently purchased a Polaron CPD and wanted to hear from anyone else who has one. I had used one at a University, at which time once I loaded the specimen chamber and shut the back, opened the fill valve, the CO2 leaked out the front window. I was concerned and drained out the CO2. I went to find the instructor, and asked why this was happening. I assumed a seal was faulty, but she told me that the boat wasn't loaded properly. The back had closed nicely, and I really didn't think that it was not loaded properly. Once the chamber was reloaded, the CO2 tank was empty and I could not finish the run. Once the CO2 tank was replaced weeks later, I received a phone call saying that the seal was damaged and that I would have to wait before the new ones came in. To make a long story short, we now have a Polaron CPD and I am having the same problem. Sometimes when I load it, it leaks out the front. Everything seems to be fitting properly, but on occasion, upon filling, it leaks out the front window. The only reason that I am posting this question to the listserver and not to the manufacturer, is because another user thinks that this is normal?! Even if the boat was not fitting properly, should the vessel still leak out the front? I would appreciate any enlightment on this problem. It is a brandnew CPD and thus I have my doubts that the seal would be damaged already. Perhaps it is just not seated properly.
P.S. If anyone has recently purchased a Polaron CPD and finds out that the seal inside the chamber door keeps falling out, a piece of teflon tape around the seal works wonders!
Susan
Susan Carbyn Atlantic Food and Horticulture Research Centre Agriculture and Agri-Food Canada Kentville, Nova Scotia B4N 1J5 Canada
Can anyone please comment on the storage properties of common EM/LM fixatives. We have some older 1% glut/4% formaldehyde in PO4 buffer. Is there some rule of thumb that people are using, should we analyze before use, are there some references we can access? Thank you in advance for your comments.
} Now we find that the university is making a Master Plan, the draft of which } shows a major service road to a new 1000 space parking deck coming very } close to our building. We estimate that the road will be within 50 feet, } or even closer, to the below-ground EM lab. Given our existing problem } with skate boarders, we are very worried that this road will essentially } destroy the utility of the lab. } Dear Candice et al., This sounds like a real disaster in the making. Our facility is several hundred meters from some major roads, and we were quite worried about vibrations from truck traffic. The effects of traffic depend crit- ically on the nature of the soil between you and the road. Bedrock will transmit vibrations very well; whereas damp clay will absorb much of the energy. The good news is that there may not be much traffic except for a few times during the day, and that you may be able to convince the university to put some vibration-damping material at the bottom of the roadbed. I don't know what is available, but maybe a layer of poly- urethane (which is a good vibration absorber) could be cost-effective solution--especially if there is a source of recycled plastic locally. Good luck. Yours, Bill Tivol
As a result of my comments on bremsstrahlung, inelastic, and elastic scattering, questions have been raised concerning energy loss as an electron's path is changed. That is,it gets swung around the nucleus somewhat like a comet gets swung around the sun, and since this involves a change in direction, which in turn amounts to deacceleration, should involve some loss in energy. As I hope I implied in my original comments, I do not consider myslef to be an authority on matters of electron-atom interactions. All I intended to do was to give a couple, what I considered to be clear and useful, references on the subject. And so, in an attempt to answer the more recent question I quote from Reimer, p. 73 of his book 'Scasnning Electron Microscopy':
"During elastic scattering of electrons, as discussed in Sect. 3.1, the sums of momenta and of kinetic energy of the collision partners are conserved. The energy loss of the incident electrons and the kinetic energy transferred to the nucleus can be neglected for the electron energies used in SEM since the electron mass is so much smaller than thet of the nucleus. Even when 30 keV electrons are scattered through an angle of 180=B0, the energy transferred to a Cu nucleus is only of the order of one electronvolt, and such scattering processes have a much lower probability than inelastic processes with energy losses larger than 5 eV. Only for electrons in the MeV region can the energy transferred bvecome larger than the displacement energy necessary to dislodge an atom from its lattice site into an interstitial position, which is of the order of 10-30 eV."
I hope this will help clarify the matter. For more details refer to Reimer's book.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
Candice/all: We here at Dow are in a well-isolated facility which is a result of demonstrating that passing trucks would give problems to our NMR spectrometers and microscopes. Without going into the full explanation, the argument was made based on empirical data: We had large trucks rumble by our existing facilities during data acquisition and compared the results to the same experiment run during a known quiet time. The loss of information was documented and recast in terms of monetary cost for reduced data quality. In our case, the financial penalty of reduced resolution/sensitivity justified the extra cost of closing a major local thoroughfare.
My suggestion would be to get someone from a trucking company to come by and drive their truck in the approximate location of the service drive to document the problems, then see if the U. can come up with an alternate access route to the ramp (moving the entire ramp would be better, but probably less likely). The fact that you have a few hundred thousand dollars tied up in sensitive equipment suggests that the U. recognizes the value of your work and would hopefully be willing to accommodate the situation. Consider the bad press they would get for compromising their research reputation in the name of a car park!
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } Hi Susan, Try filling the chamber very slowly. I have the same problem on my BioRad CPD. Changing the seals didn't help.
} Hi there, } } We have recently purchased a Polaron CPD and wanted to hear from } anyone else who has one. I had used one at a University, at which time } once I loaded the specimen chamber and shut the back, opened the fill } valve, the CO2 leaked out the front window. I was concerned and drained } out the CO2. I went to find the instructor, and asked why this was } happening. I assumed a seal was faulty, but she told me that the boat } wasn't loaded properly. The back had closed nicely, and I really didn't } think that it was not loaded properly. Once the chamber was reloaded, } the CO2 tank was empty and I could not finish the run. Once the CO2 } tank was replaced weeks later, I received a phone call saying that the seal } was damaged and that I would have to wait before the new ones came in. } To make a long story short, we now have a Polaron CPD and I am having } the same problem. Sometimes when I load it, it leaks out the front. } Everything seems to be fitting properly, but on occasion, upon filling, it } leaks out the front window. The only reason that I am posting this } question to the listserver and not to the manufacturer, is because another } user thinks that this is normal?! Even if the boat was not fitting properly, } should the vessel still leak out the front? I would appreciate any } enlightment on this problem. It is a brandnew CPD and thus I have my } doubts that the seal would be damaged already. Perhaps it is just not } seated properly. } } } } Gregory Rudomen Greg-at-UMIC.SUNYSB.EDU 516-444-3126 University Microscopy Imaging Center S.U.N.Y. Stony Brook
Contact Dr. Judy Murphy (expert in design of EM labs). She may be able to guide you. 209 474-5284
Also check Chapter 1, Setting Up An Electron Microscope Facility in Procedures in Electron Microscopy, AW Robards and AJ Wilson, eds, John Wiley & Sons, New York. While it doesn't address skateboarders and roads per se, it may give you ammunition to fight the administration.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
I am currently working on gold labelling project involving monolayr. I use LR GOLD and LOWICRYL and do my embedding and polymerization in the UVC 2 cryochamber by TED PELLA. My problem is that the blocks polymerize with in 1 hr. Has anyone experienced this? What can I do about it. I want slow polymerization.
I follow the protcols provided and at minus 10 C.
******************************************************** * Raj Patel * * Dept. of Pathology * * Robert Wood Johnson Medical School * * 675 Hoes Lane, Piscataway, NJ 08854 * * * * voice (908) 235-4648; Fax -4825 * * E-Mail rpatel-at-umdnj.edu * ********************************************************
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
ed
} } Hi there, } } We have recently purchased a Polaron CPD and wanted to hear from } anyone else who has one. I had used one at a University, at which time } once I loaded the specimen chamber and shut the back, opened the fill } valve, the CO2 leaked out the front window. I was concerned and drained } out the CO2. I went to find the instructor, and asked why this was } happening. I assumed a seal was faulty, but she told me that the boat } wasn't loaded properly. The back had closed nicely, and I really didn't } think that it was not loaded properly. Once the chamber was reloaded, } the CO2 tank was empty and I could not finish the run. Once the CO2 } tank was replaced weeks later, I received a phone call saying that the seal } was damaged and that I would have to wait before the new ones came in. } To make a long story short, we now have a Polaron CPD and I am having } the same problem. Sometimes when I load it, it leaks out the front. } Everything seems to be fitting properly, but on occasion, upon filling, it } leaks out the front window. The only reason that I am posting this } question to the listserver and not to the manufacturer, is because another } user thinks that this is normal?! Even if the boat was not fitting properly, } should the vessel still leak out the front? I would appreciate any } enlightment on this problem. It is a brandnew CPD and thus I have my } doubts that the seal would be damaged already. Perhaps it is just not } seated properly. } } P.S. If anyone has recently purchased a Polaron CPD and finds out that } the seal inside the chamber door keeps falling out, a piece of teflon tape } around the seal works wonders! } } } Susan } } } Susan Carbyn } Atlantic Food and Horticulture Research Centre } Agriculture and Agri-Food Canada } Kentville, Nova Scotia B4N 1J5 } Canada } } Phone: (902) 679-5566 } Fax: (902) 679-2311 } E-mail: carbyns-at-em.agr.ca
Ed Basgall, PhD Penn State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618
Job description: SEM operation and sample preparation in support of DRAM fabrication and failure analysis. The candidate will perform: 1. Delayering/deprocessing by RIE/Plasma, wet etch and parallel polishing/face-lapping. The deprocessing will require the use of chemicals (acids and solvents) within a wet lab. 2. Optical microscope inspections by bright field and differential interface contrast. 3. Cross section preparation using mechanical polishing, fracturing, and focused ion beam (FIB) techniques. 4. SEM inspections by secondary electron imaging, back-scatter imaging and energy dispersive spectroscopy (EDS). Education and experience: The candidate should ideally have an associates degree and several years of semiconductor SEM experience. If there is no semiconductor SEM experience, the candidate should possess a college degree in a microscopy related field (i.e. biology/geology/materials science/etc) and be actively using SEM and optical microscopy techniques. Good eye-hand coordination, communication skills, and attention to details are required. Candidates should be highly motivated, self directed, interested in learning new skills an effective working alone or in a team environment. ******************************************* Bart Seefeldt White Oak Semiconductor 600 East Main Street, Suite 800 Richmond, VA 23219 Ph: 804-698-7225 Fax: 804-698-7316 E-mail: seefeldb-at-whiteoaksemi.com *******************************************
Has anyone had experience using a Pixera camera on a TEM? I am attempting to put one on a Zeiss 902 with a C mount connection. If the camera is directly mounted, the focal length is not correct. Does anyone know of an adapter than can be used so theimage can be focused correctly?
Thanks for any suggestions.
Nancy R. Smith Microscope and Graphic Imaging Center California State University, Hayward
} } We have recently purchased a Polaron CPD and wanted to hear from } anyone else who has one. I had used one at a University, at which time } once I loaded the specimen chamber and shut the back, opened the fill } valve, the CO2 leaked out the front window. I was concerned and drained } out the CO2. I went to find the instructor, and asked why this was } happening. I assumed a seal was faulty, but she told me that the boat } wasn't loaded properly. The back had closed nicely, and I really didn't } think that it was not loaded properly. Once the chamber was reloaded, } the CO2 tank was empty and I could not finish the run. Once the CO2 } tank was replaced weeks later, I received a phone call saying that the seal } was damaged and that I would have to wait before the new ones came in. } To make a long story short, we now have a Polaron CPD and I am having } the same problem. Sometimes when I load it, it leaks out the front. } Everything seems to be fitting properly, but on occasion, upon filling, it } leaks out the front window. The only reason that I am posting this } question to the listserver and not to the manufacturer, is because another } user thinks that this is normal?!
This is EXACTLY THE SORT OF MATTER TO PUT ON THE LISTSERVER.
Even if the boat was not fitting properly, } should the vessel still leak out the front? I would appreciate any } enlightment on this problem. It is a brandnew CPD and thus I have my } doubts that the seal would be damaged already.
We have a Polaron CPD 3000 and the seals often leak, particularly the one on the window. I have taken the CPD apart myself to check it. The window seal leaks even though there are no physical defects on it. And you can replace the same seal in the window and next time it will not leak, indicating no permanent damage. So it is not damaged in the sense that a badly loaded boat has pushed up against it and nicked it so it leaks. In fact I can't see how loading the boat would ever make a difference to the integrity of either of the seals, considering how they are located completely inside grooves.
BUT the door seal can be physically damaged by hard particles (say bits of cover slip) getting washed out of the chamber and locating at the sealing surface so they nick the seal as you screw up the door. SO you need to go carefully around the DOOR sealing surface and screw thread with say ethanol on a cotton bud once a week.
AND the seals on the drain valve on the bottom need regular cleaning for the same reason. Grit washes down the drain and scores the sealing surface and O rings in the valve. In fact if you have never done it, go and clean the door seal and drain valve right away. You'll be surprised at the gunge that will be there.
My explanation is to do with seal elasticity. When the CPD is cooled before being pressurised, the neoprene seals lose much of their elasticity and do not expand properly to seal when the chamber is pressurised. So the solvent leaks out and as the seal stays cold it will never seal properly as of course you keep the chamber cold to ensure fast fluid transfer.
The remedy according to Me is to pressurise the CPD BEFORE COOLING IT. BTW we heat and cool the CPD by circulating water through the jacket using a waterbath heater/mixer (Lauda type T, -20deg to 100 deg C.) which has a small centrifugal pump on it and which sits in a 2 Litre stainless tank. At the start we fill up the small tank with ice and add enough water to cover the heater/pump on the mixer. Then we start the pump with the heater off and chill the CPD that way. When we want to warm the CPD we toss away the ice water, replace it with tap water -at- 20 deg or so and turn on the heater which warms the waterbath towards 50 deg.
Notwithstanding all of the above, since the CPD only works if all the seals are good, you must keep a couple of seal kits in your lab ready for quick repairs. Failure of the seals is usually discovered after some poor soul has spent weeks on their experiment and days on processing their tissue only to find the CPD leaks. If you have the capacity to quickly replace any of the seals so their experiment survives your reputation will be much enhanced!
Mel Dickson E.M. Unit, University of NSW Sydney, Australia
} We solicit your help in: } (1) Sharing knowledge about similar situations
I used to have a lab 50 metres from a (not so busy) railway line. We could detect the vibration from the trains when they were around a kilometre off.
The problem will vary according to the soil type between your lab and the new road. Ideally the road would be on swampy ground and your lab on a platfrom carved out of granite. That would give good decoupling. But basically, any effective solution in you laboratory will cost several thousands per instrument, as each instrument will need to be relocated on some heavy support (thick steel plate, thicker concrete slab, mass is what you need) with some very flexible mounts under it (air-springs are ideal). It may be more cost effective to route the road further away.
} (2) Pointing us to the best published sources about EM lab design, } particularly in regard to vibration and preferred distance from nearby roads
} (3) Pointing us to any expert EM lab design firms from whom we } might get information
The classic reference work is "Design of the Electron Microscope Laboratory" by Ronald H Alderson 1975. It is Volume 4 in "Practical Methods in Electron Microscopy" Editor Audrey M Glauert. American Elsevier ISBN 0444 1087 6. Was still available two years back whem I bought my second copy to share with the architect for my new laboratory. Pages 68-86 deal with mechanical vibration and decoupling/damping systems } } Mel Dickson } } } }
} } Can anyone please comment on the storage properties of common EM/LM fixatives. } We have some older 1% glut/4% formaldehyde in PO4 buffer. Is there some rule } of thumb that people are using, should we analyze before use, are there some } references we can access? } Thank you in advance for your comments. } } Regards, } Ken Baker } } The aldehydes oxidise to acids - formic or glutaric. The reaction is less at lower pH so is accelerated when you buffer the solution to pH7. Our rule is to buffer the fixative just before we use it and discard any buffered fixative older than a week.
Thanks to all those who responded in tracing Edington's Practical electron microscopy book. Tech Books distributes it through their distributor "Ceramic Book and literature Service (CBLS)."
A Lab6 cathode running on a Hitachi H7100 TEM (manufacturers brand) developed a bright rectanglar-shaped emmision pattern over the last 50-100 hours of operation, although it can be biased back to a small circular spot. We hadnt seen this before. The tip of the cathode shows a distinctly bar-shaped tip when viewed in a light microscope.
The cathode has been very stable over its 450 hours of life, although it seems to move a little vertically as it heats up. There is plenty of Lab6 left.
Does anyone know what causes the pattern, and what are the chances of getting back to a squarish/maltese cross type image if we, say, run it a bit hot for a while?
regards, Sally ---------------------------------------------------------------------- Sally Stowe |Email: stowe-at-rsbs.anu.edu.au Facility Coordinator |Post: ANU Electron Microscopy Unit |ANUEMU (RSBS) Ph 61 6 249 2743 |Australian National Univ. FAX 61 6 249 4891 |Canberra, http://online.anu.edu.au/EMU/home.htm |AUSTRALIA 0200
... snips } user thinks that this is normal?! Even if the boat was not fitting properly, } should the vessel still leak out the front? I would appreciate any } enlightment on this problem. It is a brandnew CPD and thus I have my } doubts that the seal would be damaged already. Perhaps it is just not } seated properly. } } P.S. If anyone has recently purchased a Polaron CPD and finds out that } the seal inside the chamber door keeps falling out, a piece of teflon tape } around the seal works wonders! } } } Susan } } } Susan Carbyn } Atlantic Food and Horticulture Research Centre } Agriculture and Agri-Food Canada } Kentville, Nova Scotia B4N 1J5 } Canada } } Phone: (902) 679-5566 } Fax: (902) 679-2311 } E-mail: carbyns-at-em.agr.ca
Well, I wouldn't consider leaking from the from window normal - get the supplier/manufacturer to sort it out.
However, it might be a handling problem rather than an equipment fault. Make sure you go through all the steps slowly, as sudden temperature changes in particular could be causing differential expansion.
I'd also be unhappy about the tape on the door seal. Although unlikely to cause problems, there is a remote chance it might. A better solution is to hold the metal seal edge on and give it a tap against a hard surface - the idea is to make it slightly oval, so it grips its seating.
Our Polaron E3000 CPD was bought in 1976 and is still going strong, wuite a lot of use too!
When you say Teflon tape around the seal, I in=magine you mean peripherally rather than through the hole in the middle?!
My experience is that the Doughty/Dowty? seal, the main metal ring plus nitrile (?), tends to split regularly with acetone, every 2-4 months or so. If everything has worked once, then assembly should be ok, it is possible for things to loosen but in my experience that is very unusual. You need to look at the inner face of the seal for any imperfection. I don't know if you get any specail tool, but I made up a steel oblong gizmo which is like a big screwdriver blade whichand gets turned gently with an adjustable wrench.
Hey! I've just realiased, we should have another party for its 21st birthday!!! We get a lot of parties around here, folks!
Many years ago JEOL News published an article on the design of the EM rooms at the John Innes Institute in the UK. As far as I can recall this dealt in some detail with vibration transmission. We based the design of our EM rooms on this and we have had no vibration problems.
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa eurc-at-giraffe.ru.ac.za - tel: +27 461 318168 - fax: +27 461 24377
Hi Candace. There was a discussion a while back on this topic which has been archived at the "Tips & Tricks" site. Go to the web address listed at the end of this message and follow the "Tips & Tricks link. Proceed to the Miscl. link and there it is at the top of the list. Good luck.
At 10:07 AM 4/3/97 +0131, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 352-392-1295 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
Looking for experienced TEM Service Engineer to service Philips 420 in Kansas City Area. Contact Pete Dondl at sylviapns-at-worldnet.att.net P & S Products, Inc.
In reply to: ================================================== I used to have a lab 50 metres from a (not so busy) railway line. We could detect the vibration from the trains when they were around a kilometre off. ================================================== There have been instances where the problem with the rail line has been less due to ground vibrations than to the creation of a transient magnetic field problem that correlated with the passage of a train. In about 1969, there was an SEM installed at a Philadelphia university near the main passenger line of AMRAK and Conrail, and the real problem was more related to the magnetic field (trains were pantograph (electrically) powered) than to vibrations. The problem was ultimately "solved" only by moving the microscope. So don't forget the magnetic field problem potential as well.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Larry's comment about knocking the seal out of 'round' reminded me - I forgot to mention in my earlier message that I do this in a large workshop vise. A gentle squeeze works a treat! (and that is not a naughty comment!). Plus, the squeeze is more controllable, if at first it doesn't work, you can go back for more.
We have a Phillips 300 TEM available to anyone who wants it. It was purchased in 1980 and completely renovated in 1994. It comes with cooling system and a standard diffusion pump instead of the original mmercury pump. It does need work on the condenser lens electronics, but otherwise works beautifully. It is free, anyone interested will just need to come and get it as we need it out asap.
Thanks!!
Cheri Owen Detroit Neurotrauma Institute Wayne State University Detroit, Mi 313-577-4648
Due to the overwhelming number of personal responses I received on my CPD question, I thought I would summarize some of what I found out.
My first reaction to all the responses, is that the Polaron CPD E3000 is used by many people in the field of EM and thus must be an excellent product.
Many responses indicated that for years, people have had no problems with their CPD's leaking. It hasn't been until fairly recently, after years of usage, that people have experienced their "Downty" seals (The seal in the window), leaking. I would imagine that after years of usage, they would need to be replaced, but I was shocked at the number of people who seem to be replacing them fairly regularly. One person stated that after several good runs (varies from 5 to 10 or more), they get a leak. Personally, I don't think 5 to 10 runs is a lot! This would suggest to me, that perhaps the seals are not as "good" quality as they used to make them?
Some suggested that temperature changes result in contraction/expansion of the seals which might explain the leaking. Another response indicated that the seals will dry up and crack easily. Since our machine and all of it's parts are brand new, the age of the seals are not a factor.
To test for leaks, a lot of people recommended pressurizing the vessel every time, prior to loading it with samples. This however, does not necessarily guarantee that when you load the chamber with your samples that it will NOT leak. One person felt that the problem was a handling one rather than an equipment fault. This would probably relate back to the temperature changes by filling a chamber too quickly that would result in differential expanding of the window Downty seal.
Many people replied to me and said that the Teflon tape around our door seal may be the problem or may cause future problems. No one said exactly why this was, other than a prompt reply I got from the manufacturer. The manufacturer thought that the tape may break off and become lodged in the drain valve. We only used a tiny piece wrapped tightly around the outside of the ring (Not around the entire circumference of the seal as some misinterpreted), but rather around the outer edge - like a piece of tape or bandaid one might wrap around a ring to make it fit their finger.
Many people have bent their door seal so that it's oval shape would hold it in place.
I appreciate all of you who responded and especially to the manufacturer! I didn't contact the manufacturer first thing, because I wanted to see if there were other people with some similar experiences that could help me "quickly" solve the problem.
The manufacturer assured me that if the boat wasn't loaded correctly, the door would not close at all. The most likely cause of the problem is from the Downty seal which is not manufactured by them, but rather purchased in. They have had batches know to be faulty and in such instances, they would discard and return to the original manufacturer. They state: if the front window leaks, there are only 2 possibilities:
1. The front is not screwed tight enough 2. The Downty seal is faulty
Finally, the manufacturer said that recently they had a batch of Downty seals which were of the wrong material and very quickly deteriorated with the dehydration solvents being used. As for the door seal needing to be bent, the business manager has requested the design team to review the way the seals are held in place and that some good news may come from the problems others have shared on this topic, on this list server!
As one colleague from the list server wrote: "Amazing the number of responses with the same problem. And we toil away thinking we're the only ones with the weird difficulties".
Thanks again to all who replied. Although my problem had an easy solution, it has given me insight on lots of other situations!
Susan
Susan Carbyn Atlantic Food and Horticulture Research Station Agriculture and Agri-Food Canada Kentville, Nova Scotia B4N 3R2 Canada
This last piece of information was passed along to me by the Manufacturer of the Polaron CPD.
Please ammend the e-mail or notify the customer with the faulty CPD that the seals used were not in fact made by DOWTY but of similar design, from another manufacturer, these in fact were of the wrong material so in practice the material was not faulty but the manufactured item itself was below spec.
Susan
Susan Carbyn Atlantic Food and Horticulture Research Station Agriculture and Agri-Food Canada Kentville, Nova Scotia B4N 1J5 Canada
We are interested in obtaining a carbon coated grid but in a "butterfly" or folding configuration instead of a single grid. We have some powders that are radioactive and though our microscope (TEM) is already contaminated we would like to reduce the potential for further contamination. I was thinking that a butterfly grid with carbon on both sides may help.
Any thoughts? Does anyone sell such a beast? Any other ideas for reducing the amount of particles that may come off?
Most of my work is with metal samples and I haven't worked with carbon coated grids much. Can we make something like that easily? I saw the discussions about holey carbon films but that is not quite what we are after!
TIA
John Vetrano Pacific Northwest National Laboratory Richland, WA 99352 js_vetrano-at-pnl.gov
We are in the process of purchasing a TEM for biological application. The models we are planning to check on are Hitachi H-7500, JEOL JEM-1220, and Philips CM100 BioTWIN. We would like to hear the opinions of people who have experience with these models. What are the pros and cons? Thank you very much.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Angus:
I posed your question to Bernie Kestel from Argonne National Laboratory and he offered the following:
" RE} selective etch for silicon 4/4/97
I once used the following solution on Si with a silicon oxide layer. Only the Si was polished away on the South Bay Technology 550 B jet polisher. 60 ml. HF, 90 ml. sulphuric acid, 100 ml. butyl cellosolve (2-butoxyethanol), 500 ml. methanol. Conditions: 80 volts, 50 ma., -45 degrees Centigrade, (dry ice/methanol), slightly "slow" pump speed ~ 4. Used green LED light source. 150 micron test hole to set auto trip at 6.5 on front panel by adjusting detector bias (rear knob), to get those conditions. If silicon nitride is conductive, this may not work."
You can get additional information on the Jet Polisher (Now Model 550D) on our web site at http://www.southbaytech.com.
DISCLAIMER: As we do manufacture the Model 550D Single Vertical Electropolisher, I obviously have a vested interest in promoting its use.
David Henriks TEL: 800-728-2233 (toll-free in USA) South Bay Technology, Inc. 714-492-2600 1120 Via Callejon FAX: 714-492-1499 San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
To any who can help...... I need to find a way to project some 4x5 negatives onto a screen for quantitation purposes. I know that such projectors used to be around, but here at Mayo they are history. I need the projector because I have many low mag micrographs that I am quantitating, and making larger (16x20) prints is not really feasable. Any leads as to where I can borrow, buy or make such a projector would be very helpful!!!
TIA Eugene Krueger GI Research Mayo Foundation krueger.eugene-at-mayo.edu
} We are interested in obtaining a carbon coated grid but in a "butterfly" or } folding configuration instead of a single grid. We have some powders that are } radioactive and though our microscope (TEM) is already contaminated we would } like to reduce the potential for further contamination. I was thinking that a } butterfly grid with carbon on both sides may help. } } Any thoughts? Does anyone sell such a beast?
I haven't seen this advertised, but one of the suppliers on our list will certainly correct me if they sell these.
} Any other ideas for reducing the } amount of particles that may come off? } A formvar or collodion film could work--especially if charging is not a problem. You don't say whether the sample is conductive or not.
} Most of my work is with metal samples and I haven't worked with carbon coated } grids much. Can we make something like that easily?
Yes, [even I can make one :-)]. Two procedures are possible: 1) Cleave a mica sheet to expose a fresh surface, evaporate carbon onto that surface, lower the carbon-coated mica at an angle into a staining dish filled with distilled water to float the film off the mica
Prepare the grids by rinsing in dilute nitric acid then distilled water, place open folding grids, inside down, onto the floating carbon film (care- fully), place a square of filter paper over the grids & film and lift off the surface of the water (again, carefully). 2) Buy or make a solution of formvar in ethylene dichloride--the proper dilution will depend on the final thickness of the film. Take a clean glass microscope slide and apply a very light coating of finger or nose grease. Dip the slide into the solution, let the excess drip off and let the solvent evaporate in dry air--if you are in a humid environment, you will have to fill a volume with dry N2. When the film has hardened, score the slide with a razor blade very near the edges to give a film which is attached only to one surface of the slide. Lower the slide at an angle into 60 deg C water to float the film off, prepare, place & pick up grids as above. Evaporate carbon onto the formvar-coated grids. The first method gives a thinner film, but it may not prevent the escape of small bits of powder; the second method gives a less porous film. When you want to use the grids, you will have to cut them out to remove them from the filter paper. Be careful to make sure that the film adheres to the whole surface of the grid during this and the folding process. Good luck. Yours, Bill Tivol
Dear Colleagues: Can any of you recommend a good reference book on preparations of macromolecules, e.g.,DNA and proteins, for electron microscopy? I am looking for a comprehensive review book which describes methods of positive, negative staining, rotary shadowing of biological macromolecules. Thank you in advance.
If I understand your question correctly, I think that you have several options.
1) You could make your own carbon coated folding grids using the same procedures used for single grids ( check for example "Techniques for Electron Microscopy" Ed. Desmond Kay ). The techniques (there are a number of them) are relatively simple.
2) Could you use a single carbon coated grid and then deposit carbon (by vacuum evaporation) on top of your particles on the grid ?. This would serve the same purpose as the carbon coated folding grids. You might end up contaminating your evaporator however.
No matter what you do, you will still run the risk of contaminating your scope since some of the film might break during observation. Also , keep in mind that the increased thickness (two carbon layers), will decrease your resolution. This might or might not affect the information you are after.
The question was how long fixatives can be stored in an TEM lab.
After exhaustive investigation some years ago, we arrived at the following answer: The highest grade glutaraldehyde or paraformaldehyde (distilled and stored in glass vials under inert gas) will deterioate to approximately one half their strengths in 3 weeks assuming they are in a buffer, approximately at pH 7.4, and under continuous refrigeration. Therefore we never keep fixatives for more than one week. We make them up the day of, or the day before we use them. Consider how valuable your sample is and how perfect it is expected to look. A sample which has undergone autolysis will not benefit by ultra-fresh fixation fluids, but living cells in culture will. Oxygen and heat are deleterious to aldehydes. Glutaraldehyde in a mostly empty bottle will not last very long, while glut in a glass vial sealed under inert gas will last many years. Hope this helps. Bye, Hildy
In a similar vein, can any body point me to a vender of such lenses in general. We have a Pixera that we use on the photo tube of several Olympus scopes. We borrowed the screw-in lens from the front of our Dage RS-170 camera. We get the right focal length, but the lens magnification is apparently matched to the size of an RS-170 and not a small CCD chip; therefore we get about an extra three-fold mag over what we see in the eyepieces.
We would like to find a similar lens with a mag of 1x or slightly less. Our adapter lens screws into the front of our Dage or Pixera (whatever you call that kind of mount) and has a 49 mm OD tube that slides into an adapter for our Olympus microscopes.
TIA, Warren
At 02:09 PM 4/3/97 +0000, Nancy wrote: } } Has anyone had experience using a Pixera camera on a TEM? I am } attempting to put one on a Zeiss 902 with a C mount connection. If the camera is } directly mounted, the focal length is not correct. Does anyone know } of an adapter than can be used so theimage can be focused correctly? } } Thanks for any suggestions.
--Boundary (ID jVqXAP1TDFh0bi8LAyJgag) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Susan,
} } We have recently purchased a Polaron CPD and wanted to hear from } anyone else who has one. I had used one at a University, at which time } once I loaded the specimen chamber and shut the back, opened the fill } valve, the CO2 leaked out the front window. I was concerned and drained } out the CO2. I went to find the instructor, and asked why this was } happening. I assumed a seal was faulty, but she told me that the boat } wasn't loaded properly. The back had closed nicely, and I really didn't } think that it was not loaded properly. Once the chamber was reloaded, } the CO2 tank was empty and I could not finish the run. Once the CO2 } tank was replaced weeks later, I received a phone call saying that the seal } was damaged and that I would have to wait before the new ones came in. } To make a long story short, we now have a Polaron CPD and I am having } the same problem. Sometimes when I load it, it leaks out the front. } Everything seems to be fitting properly, but on occasion, upon filling, it } leaks out the front window. The only reason that I am posting this } question to the listserver and not to the manufacturer, is because another } user thinks that this is normal?!
This is EXACTLY THE SORT OF MATTER TO PUT ON THE LISTSERVER.
Even if the boat was not fitting properly, } should the vessel still leak out the front? I would appreciate any } enlightment on this problem. It is a brandnew CPD and thus I have my } doubts that the seal would be damaged already.
We have a Polaron CPD 3000 and the seals often leak, particularly the one on the window. I have taken the CPD apart myself to check it. The window seal leaks even though there are no physical defects on it. And you can replace the same seal in the window and next time it will not leak, indicating no permanent damage. So it is not damaged in the sense that a badly loaded boat has pushed up against it and nicked it so it leaks. In fact I can't see how loading the boat would ever make a difference to the integrity of either of the seals, considering how they are located completely inside grooves.
BUT the door seal can be physically damaged by hard particles (say bits of cover slip) getting washed out of the chamber and locating at the sealing surface so they nick the seal as you screw up the door. SO you need to go carefully around the DOOR sealing surface and screw thread with say ethanol on a cotton bud once a week.
AND the seals on the drain valve on the bottom need regular cleaning for the same reason. Grit washes down the drain and scores the sealing surface and O rings in the valve. In fact if you have never done it, go and clean the door seal and drain valve right away. You'll be surprised at the gunge that will be there.
My explanation is to do with seal elasticity. When the CPD is cooled before being pressurised, the neoprene seals lose much of their elasticity and do not expand properly to seal when the chamber is pressurised. So the solvent leaks out and as the seal stays cold it will never seal properly as of course you keep the chamber cold to ensure fast fluid transfer.
The remedy according to Me is to pressurise the CPD BEFORE COOLING IT. BTW we heat and cool the CPD by circulating water through the jacket using a waterbath heater/mixer (Lauda type T, -20deg to 100 deg C.) which has a small centrifugal pump on it and which sits in a 2 Litre stainless tank. At the start we fill up the small tank with ice and add enough water to cover the heater/pump on the mixer. Then we start the pump with the heater off and chill the CPD that way. When we want to warm the CPD we toss away the ice water, replace it with tap water -at- 20 deg or so and turn on the heater which warms the waterbath towards 50 deg.
Notwithstanding all of the above, since the CPD only works if all the seals are good, you must keep a couple of seal kits in your lab ready for quick repairs. Failure of the seals is usually discovered after some poor soul has spent weeks on their experiment and days on processing their tissue only to find the CPD leaks. If you have the capacity to quickly replace any of the seals so their experiment survives your reputation will be much enhanced!
Mel Dickson E.M. Unit, University of NSW Sydney, Australia
--Boundary (ID jVqXAP1TDFh0bi8LAyJgag) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHA38D08MO002DD2-at-mc.duke.edu} ; Thu, 03 Apr 1997 19:23:42 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id TAA03810; Thu, 03 Apr 1997 19:20:34 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id RAA09492 for dist-Microscopy; Thu, 03 Apr 1997 17:26:25 -0600 Received: from sam.comms.unsw.EDU.AU (sam.comms.unsw.EDU.AU [149.171.96.20]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id RAA09489 for {microscopy-at-sparc5.microscopy.com} ; Thu, 03 Apr 1997 17:26:22 -0600 Received: from the_boss (emunit2.med.unsw.EDU.AU [149.171.168.123]) by sam.comms.unsw.EDU.AU (8.7.5/8.7.5.kenso-central) with SMTP id JAA27392 for {microscopy-at-sparc5.microscopy.com} ; Fri, 04 Apr 1997 09:37:48 +1000 (EST)
--Boundary (ID n+qYe/O3jOk1y2QtvxOlhg) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } Can anyone please comment on the storage properties of common EM/LM fixatives. } We have some older 1% glut/4% formaldehyde in PO4 buffer. Is there some rule } of thumb that people are using, should we analyze before use, are there some } references we can access? } Thank you in advance for your comments. } } Regards, } Ken Baker } } The aldehydes oxidise to acids - formic or glutaric. The reaction is less at lower pH so is accelerated when you buffer the solution to pH7. Our rule is to buffer the fixative just before we use it and discard any buffered fixative older than a week.
Mel Dickson }
--Boundary (ID n+qYe/O3jOk1y2QtvxOlhg) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHA3KBIII8003CCZ-at-mc.duke.edu} ; Thu, 03 Apr 1997 19:33:19 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id TAA04007; Thu, 03 Apr 1997 19:30:16 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id RAA09581 for dist-Microscopy; Thu, 03 Apr 1997 17:46:34 -0600 Received: from sam.comms.unsw.EDU.AU (sam.comms.unsw.EDU.AU [149.171.96.20]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id RAA09578 for {microscopy-at-sparc5.microscopy.com} ; Thu, 03 Apr 1997 17:46:31 -0600 Received: from the_boss (emunit2.med.unsw.EDU.AU [149.171.168.123]) by sam.comms.unsw.EDU.AU (8.7.5/8.7.5.kenso-central) with SMTP id JAA08217 for {microscopy-at-sparc5.microscopy.com} ; Fri, 04 Apr 1997 09:58:02 +1000 (EST)
--Boundary (ID tvuincBIJPRoJYC4i3obdg) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} We solicit your help in: } (1) Sharing knowledge about similar situations
I used to have a lab 50 metres from a (not so busy) railway line. We could detect the vibration from the trains when they were around a kilometre off.
The problem will vary according to the soil type between your lab and the new road. Ideally the road would be on swampy ground and your lab on a platfrom carved out of granite. That would give good decoupling. But basically, any effective solution in you laboratory will cost several thousands per instrument, as each instrument will need to be relocated on some heavy support (thick steel plate, thicker concrete slab, mass is what you need) with some very flexible mounts under it (air-springs are ideal). It may be more cost effective to route the road further away.
} (2) Pointing us to the best published sources about EM lab design, } particularly in regard to vibration and preferred distance from nearby roads
} (3) Pointing us to any expert EM lab design firms from whom we } might get information
The classic reference work is "Design of the Electron Microscope Laboratory" by Ronald H Alderson 1975. It is Volume 4 in "Practical Methods in Electron Microscopy" Editor Audrey M Glauert. American Elsevier ISBN 0444 1087 6. Was still available two years back whem I bought my second copy to share with the architect for my new laboratory. Pages 68-86 deal with mechanical vibration and decoupling/damping systems } } Mel Dickson } } } }
--Boundary (ID tvuincBIJPRoJYC4i3obdg) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHA3MU5RCW003010-at-mc.duke.edu} ; Thu, 03 Apr 1997 19:35:25 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id TAA04031; Thu, 03 Apr 1997 19:32:18 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id RAA09556 for dist-Microscopy; Thu, 03 Apr 1997 17:41:57 -0600 Received: from sam.comms.unsw.EDU.AU (sam.comms.unsw.EDU.AU [149.171.96.20]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id RAA09553 for {microscopy-at-sparc5.microscopy.com} ; Thu, 03 Apr 1997 17:41:54 -0600 Received: from the_boss (emunit2.med.unsw.EDU.AU [149.171.168.123]) by sam.comms.unsw.EDU.AU (8.7.5/8.7.5.kenso-central) with SMTP id JAA05587 for {microscopy-at-sparc5.microscopy.com} ; Fri, 04 Apr 1997 09:53:23 +1000 (EST)
Use a high intensity overhead projector with a cardboard mask to prevent blinding white light.
John D. Warren Area Sales Manager Digital Products Polaroid Corporation
4525 Leonard Parkway Richmond, Virginia 23221-1809
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
To any who can help...... I need to find a way to project some 4x5 negatives onto a screen for quantitation purposes. I know that such projectors used to be around, but here at Mayo they are history. I need the projector because I have many low mag micrographs that I am quantitating, and making larger (16x20) prints is not really feasable. Any leads as to where I can borrow, buy or make such a projector would be very helpful!!!
TIA Eugene Krueger GI Research Mayo Foundation krueger.eugene-at-mayo.edu
I've seen this type of "squarish" pattern before. You have most likely lost the central "tip" of your LaB6 filament. It may have fractured off leaving a truncated pyramid shape, which can give this "filament" image. I doubt if there is anything you can do at this point, as you cannot reform the tip if it is missing, however if you are still getting plenty of emission from the LaB6 then you will be okay for normal microscopy. The coherence will likely drop and high resolution imaging may degrade.
Running it "hot" will not reform the tip as you can sometimes do with a Field Emission Source
--Boundary (ID RYtLB+B2qeqzbD2h2WkqhQ) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thanks to all those who responded in tracing Edington's Practical electron microscopy book. Tech Books distributes it through their distributor "Ceramic Book and literature Service (CBLS)."
CBLS c an be contacted at 703-758-2539 (ph#)
703-758-1518 (fax#)
or
614-374-9458 (ph#) 614-374-8029 (fax#).
Mohan Kalyanaraman
--Boundary (ID RYtLB+B2qeqzbD2h2WkqhQ) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHA81UVWTS0038EC-at-mc.duke.edu} ; Thu, 03 Apr 1997 21:41:11 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id VAA06190; Thu, 03 Apr 1997 21:38:23 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id TAA10213 for dist-Microscopy; Thu, 03 Apr 1997 19:37:02 -0600 Received: from VAXC.PAU.MOBIL.COM (pau.mobil.com [131.126.24.52]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id TAA10210 for {microscopy-at-sparc5.microscopy.com} ; Thu, 03 Apr 1997 19:36:58 -0600 Received: by vaxc.pau.mobil.com (UCX V4.0-10B, OpenVMS V5.5-2 VAX); Thu, 03 Apr 1997 20:46:41 -0500 Received: from ccMail by ccmail.pau.mobil.com (SMTPLINK V2.11.01) id AA860129221; Thu, 03 Apr 1997 20:41:16 -0500 (EST)
Seminar announcement: Optimizing Light Microscopy
A one-day lecture/demonstration for anyone who uses or plans to use a microscope: students, teachers, medical technologists, clinicians, pathologists, and lab managers
Six locations: Monday, April 28 - Marriott Marquis, NYC Tuesday, April 29 - Plainview Plaza-Hotel, Plainview, NY Thursday, May 1 - Tufts Medical School/Multi-Media Resource Center, Boston, MA Friday, May 2 - Tufts Medical School/Multi-Media Resource Center, Boston, MA Tuesday, May 6 - Holidome, Holyoke, MA (Greater Springfield area) * Wednesday, May 7 - Sheraton Hartford, Hartford, CT * * catered lunch available for an additional $15.50
Program: 1. A quick tour around the microscope 2. Alignment tips for reducing headaches, fatigue, and errors 3. Care, cleaning, and troubleshooting 4. Useful principles for understanding images 5. Quick, easy, and often free techniques for improving contrast (includes discussions of phase and Hoffman Modulation Contrast) 6. Advanced techniques (Fluorescence and DIC) 7. Becoming a better consumer: matching your microscope to your applications 8. The Video connection: cameras, computers, and your microscope
Fees: $115 if received before 4/18/97; $125 if received after that date. $50 discount on tuition for third person registered from the same facility.
Free with your tuition: “Optimizing Light Microscopy for Biological and Clinical Laboratories” Over 220 pages of helpful tips, quick experiments, and new ideas for getting the best from your microscope (ASCLS/Kendall-Hunt, 1997)
For further details, contact : Dr. Kenneth Piel or Barbara Foster Microscopy/Microscopy Education (MME) 53 Eton Street Springfield, MA 01108 Phone: (413)746-6931 Fax: (413)746-9311 email: mme-at-map.com
To register: Fax or mail the form below to the MME office Name: ___________________________________________________________________ School/Hospital/Company:_________________________________________________Address: ________________________________________________________________ City/State/Zip: _________________________________________________________ Phone: ____________________Fax: _______________email:____________________ Check enclosed (payable to MME) for: _________________ [ ] tuition only [ ] tuition plus lunch [ ] VISA [ ] Master Card Name on credit card: __________________________________________________ Card #: ____________________________________ Expiration date:___________
--Boundary (ID OtZ1SQoR3F56bpZ1tehfPQ) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
... snips } user thinks that this is normal?! Even if the boat was not fitting properly, } should the vessel still leak out the front? I would appreciate any } enlightment on this problem. It is a brandnew CPD and thus I have my } doubts that the seal would be damaged already. Perhaps it is just not } seated properly. } } P.S. If anyone has recently purchased a Polaron CPD and finds out that } the seal inside the chamber door keeps falling out, a piece of teflon tape } around the seal works wonders! } } } Susan } } } Susan Carbyn } Atlantic Food and Horticulture Research Centre } Agriculture and Agri-Food Canada } Kentville, Nova Scotia B4N 1J5 } Canada } } Phone: (902) 679-5566 } Fax: (902) 679-2311 } E-mail: carbyns-at-em.agr.ca
Well, I wouldn't consider leaking from the from window normal - get the supplier/manufacturer to sort it out.
However, it might be a handling problem rather than an equipment fault. Make sure you go through all the steps slowly, as sudden temperature changes in particular could be causing differential expansion.
I'd also be unhappy about the tape on the door seal. Although unlikely to cause problems, there is a remote chance it might. A better solution is to hold the metal seal edge on and give it a tap against a hard surface - the idea is to make it slightly oval, so it grips its seating.
Regards, Larry Stoter
--Boundary (ID OtZ1SQoR3F56bpZ1tehfPQ) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHAKPYJWDC003HAR-at-mc.duke.edu} ; Fri, 04 Apr 1997 03:44:14 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id DAA10792; Fri, 04 Apr 1997 03:41:23 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id BAA11075 for dist-Microscopy; Fri, 04 Apr 1997 01:17:00 -0600 Received: from relay-7.mail.demon.net (relay-7.mail.demon.net [194.217.242.9]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id BAA11072 for {Microscopy-at-sparc5.microscopy.com} ; Fri, 04 Apr 1997 01:16:58 -0600 Received: from teknesis.demon.co.uk ([158.152.199.245]) by relay-5.mail.demon.net id aa0528981; Fri, 04 Apr 1997 08:19 -0300 (BST)
--Boundary (ID 547gv2vDQtwfAys+HAC/Pg) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello Mel and others intetested
Many years ago JEOL News published an article on the design of the EM rooms at the John Innes Institute in the UK. As far as I can recall this dealt in some detail with vibration transmission. We based the design of our EM rooms on this and we have had no vibration problems.
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa eurc-at-giraffe.ru.ac.za - tel: +27 461 318168 - fax: +27 461 24377
--Boundary (ID 547gv2vDQtwfAys+HAC/Pg) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHAMYPQXZ4002EZV-at-mc.duke.edu} ; Fri, 04 Apr 1997 04:48:33 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id EAA11249; Fri, 04 Apr 1997 04:45:45 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id CAA11216 for dist-Microscopy; Fri, 04 Apr 1997 02:37:25 -0600 Received: from quagga ([146.231.128.2]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id CAA11213 for {microscopy-at-sparc5.microscopy.com} ; Fri, 04 Apr 1997 02:37:11 -0600 Received: from giraffe.ru.ac.za [146.231.128.10] by quagga.ru.ac.za with esmtp (Exim 1.58 #1) id 0wD3Gj-0004tK-00; Fri, 04 Apr 1997 09:18:41 +0200 Received: from GIRAFFE/MERCQ by giraffe.ru.ac.za (Mercury 1.21); Fri, 04 Apr 1997 09:18:36 +0200 Received: from MERCQ by GIRAFFE (Mercury 1.21); Fri, 04 Apr 1997 08:25:02 +0200
--Boundary (ID GV24A/QzYVfNi1JegNvAxQ) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello Susan
Our Polaron E3000 CPD was bought in 1976 and is still going strong, wuite a lot of use too!
When you say Teflon tape around the seal, I in=magine you mean peripherally rather than through the hole in the middle?!
My experience is that the Doughty/Dowty? seal, the main metal ring plus nitrile (?), tends to split regularly with acetone, every 2-4 months or so. If everything has worked once, then assembly should be ok, it is possible for things to loosen but in my experience that is very unusual. You need to look at the inner face of the seal for any imperfection. I don't know if you get any specail tool, but I made up a steel oblong gizmo which is like a big screwdriver blade whichand gets turned gently with an adjustable wrench.
Hey! I've just realiased, we should have another party for its 21st birthday!!! We get a lot of parties around here, folks!
Best wishes - Keith Ryan Plymouth Marine Lab. UK
--Boundary (ID GV24A/QzYVfNi1JegNvAxQ) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHAOG9OHF40029NX-at-mc.duke.edu} ; Fri, 04 Apr 1997 05:31:00 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id FAA11456; Fri, 04 Apr 1997 05:28:08 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id CAA11150 for dist-Microscopy; Fri, 04 Apr 1997 02:13:18 -0600 Received: from mail.nerc-swindon.ac.uk ([192.171.191.9]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id CAA11147 for {Microscopy-at-Sparc5.Microscopy.Com} ; Fri, 04 Apr 1997 02:13:12 -0600 Received: from kwpf11.nerc-keyworth.ac.uk by mail.nerc-swindon.ac.uk with SMTP (PP) id {02484-0-at-mail.nerc-swindon.ac.uk} ; Fri, 04 Apr 1997 09:25:30 +0100 Received: from Keyworth-Message_Server by wpo.nerc.ac.uk with Novell_GroupWise; Fri, 04 Apr 1997 08:22:16 +0000
--Boundary (ID q7po8Fxrif6bAclQj9U3Wg) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Forwarded message:
--Boundary (ID b0TrP1SlIILY7AxWDtcwsA) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Looking for experienced TEM Service Engineer to service Philips 420 in Kansas City Area. Contact Pete Dondl at sylviapns-at-worldnet.att.net P & S Products, Inc.
--Boundary (ID b0TrP1SlIILY7AxWDtcwsA) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHAW788T00003AA7-at-mc.duke.edu} ; Fri, 04 Apr 1997 09:12:44 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id JAA16710; Fri, 04 Apr 1997 09:09:38 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id HAA11890 for dist-Microscopy; Fri, 04 Apr 1997 07:10:58 -0600 Received: from mtigwc03.worldnet.att.net (mailhost.worldnet.att.net [204.127.131.34]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id HAA11887 for {MICROSCOPY-at-MSA.MICROSCOPY.COM} ; Fri, 04 Apr 1997 07:10:57 -0600 Received: from default ([207.116.103.162]) by mtigwc03.worldnet.att.net (post.office MTA v2.0 0613 ) with ESMTP id AAA23516 for {MICROSCOPY-at-MSA.MICROSCOPY.COM} ; Fri, 04 Apr 1997 13:21:12 +0000
--Boundary (ID k5gEgzEf9pExiog1hhOElA) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi Candace. There was a discussion a while back on this topic which has been archived at the "Tips & Tricks" site. Go to the web address listed at the end of this message and follow the "Tips & Tricks link. Proceed to the Miscl. link and there it is at the top of the list. Good luck.
At 10:07 AM 4/3/97 +0131, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 352-392-1295 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
--Boundary (ID k5gEgzEf9pExiog1hhOElA) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHAXINOIKW002ATT-at-mc.duke.edu} ; Fri, 04 Apr 1997 09:50:57 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id JAA17862; Fri, 04 Apr 1997 09:48:05 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id HAA11873 for dist-Microscopy; Fri, 04 Apr 1997 07:06:42 -0600 Received: from snarl.biotech.ufl.edu (snarl.biotech.ufl.edu [128.227.186.119]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id HAA11870 for {microscopy-at-Sparc5.Microscopy.Com} ; Fri, 04 Apr 1997 07:06:41 -0600 Received: from scope.biotech.ufl.edu by snarl.biotech.ufl.edu (8.7/4.09) id IAA25755; Fri, 04 Apr 1997 08:12:52 -0500 (EST)
--Boundary (ID Ao4Foz7+EQF7AtsS0Z/I5Q) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
In reply to: ================================================== I used to have a lab 50 metres from a (not so busy) railway line. We could detect the vibration from the trains when they were around a kilometre off. ================================================== There have been instances where the problem with the rail line has been less due to ground vibrations than to the creation of a transient magnetic field problem that correlated with the passage of a train. In about 1969, there was an SEM installed at a Philadelphia university near the main passenger line of AMRAK and Conrail, and the real problem was more related to the magnetic field (trains were pantograph (electrically) powered) than to vibrations. The problem was ultimately "solved" only by moving the microscope. So don't forget the magnetic field problem potential as well.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
--Boundary (ID Ao4Foz7+EQF7AtsS0Z/I5Q) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHAWZM454W003DX7-at-mc.duke.edu} ; Fri, 04 Apr 1997 09:35:39 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id JAA17314; Fri, 04 Apr 1997 09:32:20 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id HAA11931 for dist-Microscopy; Fri, 04 Apr 1997 07:25:05 -0600 Received: from ns1.axs2000.net (ns1.axs2000.net [207.106.10.100]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id HAA11928 for {Microscopy-at-sparc5.microscopy.com} ; Fri, 04 Apr 1997 07:25:04 -0600 Received: from [205.186.65.46] (tfx-us7-14.ix.netcom.com [205.186.65.46]) by ns1.axs2000.net (8.6.13/8.6.12) with SMTP id IAA05363 for {Microscopy-at-MSA.Microscopy.com} ; Fri, 04 Apr 1997 08:15:43 -0500
--Boundary (ID RFlJbowy/g7mfn4tw5qA0w) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Susan,
} } We have recently purchased a Polaron CPD and wanted to hear from } anyone else who has one. I had used one at a University, at which time } once I loaded the specimen chamber and shut the back, opened the fill } valve, the CO2 leaked out the front window. I was concerned and drained } out the CO2. I went to find the instructor, and asked why this was } happening. I assumed a seal was faulty, but she told me that the boat } wasn't loaded properly. The back had closed nicely, and I really didn't } think that it was not loaded properly. Once the chamber was reloaded, } the CO2 tank was empty and I could not finish the run. Once the CO2 } tank was replaced weeks later, I received a phone call saying that the seal } was damaged and that I would have to wait before the new ones came in. } To make a long story short, we now have a Polaron CPD and I am having } the same problem. Sometimes when I load it, it leaks out the front. } Everything seems to be fitting properly, but on occasion, upon filling, it } leaks out the front window. The only reason that I am posting this } question to the listserver and not to the manufacturer, is because another } user thinks that this is normal?!
This is EXACTLY THE SORT OF MATTER TO PUT ON THE LISTSERVER.
Even if the boat was not fitting properly, } should the vessel still leak out the front? I would appreciate any } enlightment on this problem. It is a brandnew CPD and thus I have my } doubts that the seal would be damaged already.
We have a Polaron CPD 3000 and the seals often leak, particularly the one on the window. I have taken the CPD apart myself to check it. The window seal leaks even though there are no physical defects on it. And you can replace the same seal in the window and next time it will not leak, indicating no permanent damage. So it is not damaged in the sense that a badly loaded boat has pushed up against it and nicked it so it leaks. In fact I can't see how loading the boat would ever make a difference to the integrity of either of the seals, considering how they are located completely inside grooves.
BUT the door seal can be physically damaged by hard particles (say bits of cover slip) getting washed out of the chamber and locating at the sealing surface so they nick the seal as you screw up the door. SO you need to go carefully around the DOOR sealing surface and screw thread with say ethanol on a cotton bud once a week.
AND the seals on the drain valve on the bottom need regular cleaning for the same reason. Grit washes down the drain and scores the sealing surface and O rings in the valve. In fact if you have never done it, go and clean the door seal and drain valve right away. You'll be surprised at the gunge that will be there.
My explanation is to do with seal elasticity. When the CPD is cooled before being pressurised, the neoprene seals lose much of their elasticity and do not expand properly to seal when the chamber is pressurised. So the solvent leaks out and as the seal stays cold it will never seal properly as of course you keep the chamber cold to ensure fast fluid transfer.
The remedy according to Me is to pressurise the CPD BEFORE COOLING IT. BTW we heat and cool the CPD by circulating water through the jacket using a waterbath heater/mixer (Lauda type T, -20deg to 100 deg C.) which has a small centrifugal pump on it and which sits in a 2 Litre stainless tank. At the start we fill up the small tank with ice and add enough water to cover the heater/pump on the mixer. Then we start the pump with the heater off and chill the CPD that way. When we want to warm the CPD we toss away the ice water, replace it with tap water -at- 20 deg or so and turn on the heater which warms the waterbath towards 50 deg.
Notwithstanding all of the above, since the CPD only works if all the seals are good, you must keep a couple of seal kits in your lab ready for quick repairs. Failure of the seals is usually discovered after some poor soul has spent weeks on their experiment and days on processing their tissue only to find the CPD leaks. If you have the capacity to quickly replace any of the seals so their experiment survives your reputation will be much enhanced!
Mel Dickson E.M. Unit, University of NSW Sydney, Australia
--Boundary (ID RFlJbowy/g7mfn4tw5qA0w) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHA38D08MO002DD2-at-mc.duke.edu} ; Thu, 03 Apr 1997 19:23:42 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id TAA03810; Thu, 03 Apr 1997 19:20:34 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id RAA09492 for dist-Microscopy; Thu, 03 Apr 1997 17:26:25 -0600 Received: from sam.comms.unsw.EDU.AU (sam.comms.unsw.EDU.AU [149.171.96.20]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id RAA09489 for {microscopy-at-sparc5.microscopy.com} ; Thu, 03 Apr 1997 17:26:22 -0600 Received: from the_boss (emunit2.med.unsw.EDU.AU [149.171.168.123]) by sam.comms.unsw.EDU.AU (8.7.5/8.7.5.kenso-central) with SMTP id JAA27392 for {microscopy-at-sparc5.microscopy.com} ; Fri, 04 Apr 1997 09:37:48 +1000 (EST)
--Boundary (ID BUBE7ji2uFPHCmaM0hOdQw) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } Can anyone please comment on the storage properties of common EM/LM fixatives. } We have some older 1% glut/4% formaldehyde in PO4 buffer. Is there some rule } of thumb that people are using, should we analyze before use, are there some } references we can access? } Thank you in advance for your comments. } } Regards, } Ken Baker } } The aldehydes oxidise to acids - formic or glutaric. The reaction is less at lower pH so is accelerated when you buffer the solution to pH7. Our rule is to buffer the fixative just before we use it and discard any buffered fixative older than a week.
Mel Dickson }
--Boundary (ID BUBE7ji2uFPHCmaM0hOdQw) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHA3KBIII8003CCZ-at-mc.duke.edu} ; Thu, 03 Apr 1997 19:33:19 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id TAA04007; Thu, 03 Apr 1997 19:30:16 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id RAA09581 for dist-Microscopy; Thu, 03 Apr 1997 17:46:34 -0600 Received: from sam.comms.unsw.EDU.AU (sam.comms.unsw.EDU.AU [149.171.96.20]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id RAA09578 for {microscopy-at-sparc5.microscopy.com} ; Thu, 03 Apr 1997 17:46:31 -0600 Received: from the_boss (emunit2.med.unsw.EDU.AU [149.171.168.123]) by sam.comms.unsw.EDU.AU (8.7.5/8.7.5.kenso-central) with SMTP id JAA08217 for {microscopy-at-sparc5.microscopy.com} ; Fri, 04 Apr 1997 09:58:02 +1000 (EST)
--Boundary (ID iFLHmEz3mYa3AN7rLJlkpw) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} We solicit your help in: } (1) Sharing knowledge about similar situations
I used to have a lab 50 metres from a (not so busy) railway line. We could detect the vibration from the trains when they were around a kilometre off.
The problem will vary according to the soil type between your lab and the new road. Ideally the road would be on swampy ground and your lab on a platfrom carved out of granite. That would give good decoupling. But basically, any effective solution in you laboratory will cost several thousands per instrument, as each instrument will need to be relocated on some heavy support (thick steel plate, thicker concrete slab, mass is what you need) with some very flexible mounts under it (air-springs are ideal). It may be more cost effective to route the road further away.
} (2) Pointing us to the best published sources about EM lab design, } particularly in regard to vibration and preferred distance from nearby roads
} (3) Pointing us to any expert EM lab design firms from whom we } might get information
The classic reference work is "Design of the Electron Microscope Laboratory" by Ronald H Alderson 1975. It is Volume 4 in "Practical Methods in Electron Microscopy" Editor Audrey M Glauert. American Elsevier ISBN 0444 1087 6. Was still available two years back whem I bought my second copy to share with the architect for my new laboratory. Pages 68-86 deal with mechanical vibration and decoupling/damping systems } } Mel Dickson } } } }
--Boundary (ID iFLHmEz3mYa3AN7rLJlkpw) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHA3MU5RCW003010-at-mc.duke.edu} ; Thu, 03 Apr 1997 19:35:25 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id TAA04031; Thu, 03 Apr 1997 19:32:18 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id RAA09556 for dist-Microscopy; Thu, 03 Apr 1997 17:41:57 -0600 Received: from sam.comms.unsw.EDU.AU (sam.comms.unsw.EDU.AU [149.171.96.20]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id RAA09553 for {microscopy-at-sparc5.microscopy.com} ; Thu, 03 Apr 1997 17:41:54 -0600 Received: from the_boss (emunit2.med.unsw.EDU.AU [149.171.168.123]) by sam.comms.unsw.EDU.AU (8.7.5/8.7.5.kenso-central) with SMTP id JAA05587 for {microscopy-at-sparc5.microscopy.com} ; Fri, 04 Apr 1997 09:53:23 +1000 (EST)
Dear Angus,
Potassium hydroxide works well for etching silicon without attacking silicon dioxide. It might also work well with the nitride as an etch stop, but I don't have any experience with that. Concentration and temperature could be varied to enhance the selectivity for Si vs nitride.
D. Clark Turner MOXTEK, Inc. 452 West 1260 North Orem, Utah 84057 (801) 225-0930 email moxtek-at-moxtek.win.net
} Dear All,
} Does anyone know of a selective etch that will etch away silicon } but not silicon nitride? I'm looking at a multilayer sample in plan } view for TEM and hope to back etch the si substrate and use the } nitride as a stop layer.
--Boundary (ID muOkbwSi0BcU2wSyjYAO6g) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
In reply to: ================================================== I used to have a lab 50 metres from a (not so busy) railway line. We could detect the vibration from the trains when they were around a kilometre off. ================================================== There have been instances where the problem with the rail line has been less due to ground vibrations than to the creation of a transient magnetic field problem that correlated with the passage of a train. In about 1969, there was an SEM installed at a Philadelphia university near the main passenger line of AMRAK and Conrail, and the real problem was more related to the magnetic field (trains were pantograph (electrically) powered) than to vibrations. The problem was ultimately "solved" only by moving the microscope. So don't forget the magnetic field problem potential as well.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
--Boundary (ID muOkbwSi0BcU2wSyjYAO6g) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHAWZM454W003DX7-at-mc.duke.edu} ; Fri, 04 Apr 1997 09:35:39 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id JAA17314; Fri, 04 Apr 1997 09:32:20 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id HAA11931 for dist-Microscopy; Fri, 04 Apr 1997 07:25:05 -0600 Received: from ns1.axs2000.net (ns1.axs2000.net [207.106.10.100]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id HAA11928 for {Microscopy-at-sparc5.microscopy.com} ; Fri, 04 Apr 1997 07:25:04 -0600 Received: from [205.186.65.46] (tfx-us7-14.ix.netcom.com [205.186.65.46]) by ns1.axs2000.net (8.6.13/8.6.12) with SMTP id IAA05363 for {Microscopy-at-MSA.Microscopy.com} ; Fri, 04 Apr 1997 08:15:43 -0500
--Boundary (ID CjRZD4quvFNQRGdc0y7kaw) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi Candace. There was a discussion a while back on this topic which has been archived at the "Tips & Tricks" site. Go to the web address listed at the end of this message and follow the "Tips & Tricks link. Proceed to the Miscl. link and there it is at the top of the list. Good luck.
At 10:07 AM 4/3/97 +0131, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 352-392-1295 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
--Boundary (ID CjRZD4quvFNQRGdc0y7kaw) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHAXINOIKW002ATT-at-mc.duke.edu} ; Fri, 04 Apr 1997 09:50:57 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id JAA17862; Fri, 04 Apr 1997 09:48:05 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id HAA11873 for dist-Microscopy; Fri, 04 Apr 1997 07:06:42 -0600 Received: from snarl.biotech.ufl.edu (snarl.biotech.ufl.edu [128.227.186.119]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id HAA11870 for {microscopy-at-Sparc5.Microscopy.Com} ; Fri, 04 Apr 1997 07:06:41 -0600 Received: from scope.biotech.ufl.edu by snarl.biotech.ufl.edu (8.7/4.09) id IAA25755; Fri, 04 Apr 1997 08:12:52 -0500 (EST)
Assuming that not the entire film area is required, the alternative is to make contact prints of the required areas. 35mm ortho film or if you rather use a slightly larger format, cut up TEM sheetfilm to suit super 35 mm mounts. Make sure the copying is done emulsion to emulsion and use an enlarger as your lightsource. Once the method is established its quite fast. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 300+ Links, MSDS ************************ http://www.proscitech.com.au
} I need to find a way to project some 4x5 negatives onto a screen for } quantitation purposes. I need the projector because I have many low } mag micrographs that I am quantitating, and making larger (16x20) prints is } not really feasable. } Eugene Krueger } GI Research } Mayo Foundation } krueger.eugene-at-mayo.edu
--Boundary (ID nbW+uA+aakrajPNjIFa3Og) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Looking for experienced TEM Service Engineer to service Philips 420 in Kansas City Area. Contact Pete Dondl at sylviapns-at-worldnet.att.net P & S Products, Inc.
--Boundary (ID nbW+uA+aakrajPNjIFa3Og) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHAW788T00003AA7-at-mc.duke.edu} ; Fri, 04 Apr 1997 09:12:44 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id JAA16710; Fri, 04 Apr 1997 09:09:38 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id HAA11890 for dist-Microscopy; Fri, 04 Apr 1997 07:10:58 -0600 Received: from mtigwc03.worldnet.att.net (mailhost.worldnet.att.net [204.127.131.34]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id HAA11887 for {MICROSCOPY-at-MSA.MICROSCOPY.COM} ; Fri, 04 Apr 1997 07:10:57 -0600 Received: from default ([207.116.103.162]) by mtigwc03.worldnet.att.net (post.office MTA v2.0 0613 ) with ESMTP id AAA23516 for {MICROSCOPY-at-MSA.MICROSCOPY.COM} ; Fri, 04 Apr 1997 13:21:12 +0000
--Boundary (ID r3A3b7i5lOnpMDsF4ETQcw) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Susan,
} } We have recently purchased a Polaron CPD and wanted to hear from } anyone else who has one. I had used one at a University, at which time } once I loaded the specimen chamber and shut the back, opened the fill } valve, the CO2 leaked out the front window. I was concerned and drained } out the CO2. I went to find the instructor, and asked why this was } happening. I assumed a seal was faulty, but she told me that the boat } wasn't loaded properly. The back had closed nicely, and I really didn't } think that it was not loaded properly. Once the chamber was reloaded, } the CO2 tank was empty and I could not finish the run. Once the CO2 } tank was replaced weeks later, I received a phone call saying that the seal } was damaged and that I would have to wait before the new ones came in. } To make a long story short, we now have a Polaron CPD and I am having } the same problem. Sometimes when I load it, it leaks out the front. } Everything seems to be fitting properly, but on occasion, upon filling, it } leaks out the front window. The only reason that I am posting this } question to the listserver and not to the manufacturer, is because another } user thinks that this is normal?!
This is EXACTLY THE SORT OF MATTER TO PUT ON THE LISTSERVER.
Even if the boat was not fitting properly, } should the vessel still leak out the front? I would appreciate any } enlightment on this problem. It is a brandnew CPD and thus I have my } doubts that the seal would be damaged already.
We have a Polaron CPD 3000 and the seals often leak, particularly the one on the window. I have taken the CPD apart myself to check it. The window seal leaks even though there are no physical defects on it. And you can replace the same seal in the window and next time it will not leak, indicating no permanent damage. So it is not damaged in the sense that a badly loaded boat has pushed up against it and nicked it so it leaks. In fact I can't see how loading the boat would ever make a difference to the integrity of either of the seals, considering how they are located completely inside grooves.
BUT the door seal can be physically damaged by hard particles (say bits of cover slip) getting washed out of the chamber and locating at the sealing surface so they nick the seal as you screw up the door. SO you need to go carefully around the DOOR sealing surface and screw thread with say ethanol on a cotton bud once a week.
AND the seals on the drain valve on the bottom need regular cleaning for the same reason. Grit washes down the drain and scores the sealing surface and O rings in the valve. In fact if you have never done it, go and clean the door seal and drain valve right away. You'll be surprised at the gunge that will be there.
My explanation is to do with seal elasticity. When the CPD is cooled before being pressurised, the neoprene seals lose much of their elasticity and do not expand properly to seal when the chamber is pressurised. So the solvent leaks out and as the seal stays cold it will never seal properly as of course you keep the chamber cold to ensure fast fluid transfer.
The remedy according to Me is to pressurise the CPD BEFORE COOLING IT. BTW we heat and cool the CPD by circulating water through the jacket using a waterbath heater/mixer (Lauda type T, -20deg to 100 deg C.) which has a small centrifugal pump on it and which sits in a 2 Litre stainless tank. At the start we fill up the small tank with ice and add enough water to cover the heater/pump on the mixer. Then we start the pump with the heater off and chill the CPD that way. When we want to warm the CPD we toss away the ice water, replace it with tap water -at- 20 deg or so and turn on the heater which warms the waterbath towards 50 deg.
Notwithstanding all of the above, since the CPD only works if all the seals are good, you must keep a couple of seal kits in your lab ready for quick repairs. Failure of the seals is usually discovered after some poor soul has spent weeks on their experiment and days on processing their tissue only to find the CPD leaks. If you have the capacity to quickly replace any of the seals so their experiment survives your reputation will be much enhanced!
Mel Dickson E.M. Unit, University of NSW Sydney, Australia
--Boundary (ID r3A3b7i5lOnpMDsF4ETQcw) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHA38D08MO002DD2-at-mc.duke.edu} ; Thu, 03 Apr 1997 19:23:42 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id TAA03810; Thu, 03 Apr 1997 19:20:34 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id RAA09492 for dist-Microscopy; Thu, 03 Apr 1997 17:26:25 -0600 Received: from sam.comms.unsw.EDU.AU (sam.comms.unsw.EDU.AU [149.171.96.20]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id RAA09489 for {microscopy-at-sparc5.microscopy.com} ; Thu, 03 Apr 1997 17:26:22 -0600 Received: from the_boss (emunit2.med.unsw.EDU.AU [149.171.168.123]) by sam.comms.unsw.EDU.AU (8.7.5/8.7.5.kenso-central) with SMTP id JAA27392 for {microscopy-at-sparc5.microscopy.com} ; Fri, 04 Apr 1997 09:37:48 +1000 (EST)
--Boundary (ID V6OK1HQ0rvIR5mcHZmdUqw) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } Can anyone please comment on the storage properties of common EM/LM fixatives. } We have some older 1% glut/4% formaldehyde in PO4 buffer. Is there some rule } of thumb that people are using, should we analyze before use, are there some } references we can access? } Thank you in advance for your comments. } } Regards, } Ken Baker } } The aldehydes oxidise to acids - formic or glutaric. The reaction is less at lower pH so is accelerated when you buffer the solution to pH7. Our rule is to buffer the fixative just before we use it and discard any buffered fixative older than a week.
Mel Dickson }
--Boundary (ID V6OK1HQ0rvIR5mcHZmdUqw) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHA3KBIII8003CCZ-at-mc.duke.edu} ; Thu, 03 Apr 1997 19:33:19 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id TAA04007; Thu, 03 Apr 1997 19:30:16 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id RAA09581 for dist-Microscopy; Thu, 03 Apr 1997 17:46:34 -0600 Received: from sam.comms.unsw.EDU.AU (sam.comms.unsw.EDU.AU [149.171.96.20]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id RAA09578 for {microscopy-at-sparc5.microscopy.com} ; Thu, 03 Apr 1997 17:46:31 -0600 Received: from the_boss (emunit2.med.unsw.EDU.AU [149.171.168.123]) by sam.comms.unsw.EDU.AU (8.7.5/8.7.5.kenso-central) with SMTP id JAA08217 for {microscopy-at-sparc5.microscopy.com} ; Fri, 04 Apr 1997 09:58:02 +1000 (EST)
--Boundary (ID j7lIKE0hoMhSTwqeqQja/g) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} We solicit your help in: } (1) Sharing knowledge about similar situations
I used to have a lab 50 metres from a (not so busy) railway line. We could detect the vibration from the trains when they were around a kilometre off.
The problem will vary according to the soil type between your lab and the new road. Ideally the road would be on swampy ground and your lab on a platfrom carved out of granite. That would give good decoupling. But basically, any effective solution in you laboratory will cost several thousands per instrument, as each instrument will need to be relocated on some heavy support (thick steel plate, thicker concrete slab, mass is what you need) with some very flexible mounts under it (air-springs are ideal). It may be more cost effective to route the road further away.
} (2) Pointing us to the best published sources about EM lab design, } particularly in regard to vibration and preferred distance from nearby roads
} (3) Pointing us to any expert EM lab design firms from whom we } might get information
The classic reference work is "Design of the Electron Microscope Laboratory" by Ronald H Alderson 1975. It is Volume 4 in "Practical Methods in Electron Microscopy" Editor Audrey M Glauert. American Elsevier ISBN 0444 1087 6. Was still available two years back whem I bought my second copy to share with the architect for my new laboratory. Pages 68-86 deal with mechanical vibration and decoupling/damping systems } } Mel Dickson } } } }
--Boundary (ID j7lIKE0hoMhSTwqeqQja/g) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHA3MU5RCW003010-at-mc.duke.edu} ; Thu, 03 Apr 1997 19:35:25 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id TAA04031; Thu, 03 Apr 1997 19:32:18 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id RAA09556 for dist-Microscopy; Thu, 03 Apr 1997 17:41:57 -0600 Received: from sam.comms.unsw.EDU.AU (sam.comms.unsw.EDU.AU [149.171.96.20]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id RAA09553 for {microscopy-at-sparc5.microscopy.com} ; Thu, 03 Apr 1997 17:41:54 -0600 Received: from the_boss (emunit2.med.unsw.EDU.AU [149.171.168.123]) by sam.comms.unsw.EDU.AU (8.7.5/8.7.5.kenso-central) with SMTP id JAA05587 for {microscopy-at-sparc5.microscopy.com} ; Fri, 04 Apr 1997 09:53:23 +1000 (EST)
--Boundary (ID 7NVGMqAXRF2hIzZl4N5/rw) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thanks to all those who responded in tracing Edington's Practical electron microscopy book. Tech Books distributes it through their distributor "Ceramic Book and literature Service (CBLS)."
CBLS c an be contacted at 703-758-2539 (ph#)
703-758-1518 (fax#)
or
614-374-9458 (ph#) 614-374-8029 (fax#).
Mohan Kalyanaraman
--Boundary (ID 7NVGMqAXRF2hIzZl4N5/rw) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHA81UVWTS0038EC-at-mc.duke.edu} ; Thu, 03 Apr 1997 21:41:11 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id VAA06190; Thu, 03 Apr 1997 21:38:23 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id TAA10213 for dist-Microscopy; Thu, 03 Apr 1997 19:37:02 -0600 Received: from VAXC.PAU.MOBIL.COM (pau.mobil.com [131.126.24.52]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id TAA10210 for {microscopy-at-sparc5.microscopy.com} ; Thu, 03 Apr 1997 19:36:58 -0600 Received: by vaxc.pau.mobil.com (UCX V4.0-10B, OpenVMS V5.5-2 VAX); Thu, 03 Apr 1997 20:46:41 -0500 Received: from ccMail by ccmail.pau.mobil.com (SMTPLINK V2.11.01) id AA860129221; Thu, 03 Apr 1997 20:41:16 -0500 (EST)
--Boundary (ID uHV94w9QrtOCr/FV43L4kg) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Susan,
} } We have recently purchased a Polaron CPD and wanted to hear from } anyone else who has one. I had used one at a University, at which time } once I loaded the specimen chamber and shut the back, opened the fill } valve, the CO2 leaked out the front window. I was concerned and drained } out the CO2. I went to find the instructor, and asked why this was } happening. I assumed a seal was faulty, but she told me that the boat } wasn't loaded properly. The back had closed nicely, and I really didn't } think that it was not loaded properly. Once the chamber was reloaded, } the CO2 tank was empty and I could not finish the run. Once the CO2 } tank was replaced weeks later, I received a phone call saying that the seal } was damaged and that I would have to wait before the new ones came in. } To make a long story short, we now have a Polaron CPD and I am having } the same problem. Sometimes when I load it, it leaks out the front. } Everything seems to be fitting properly, but on occasion, upon filling, it } leaks out the front window. The only reason that I am posting this } question to the listserver and not to the manufacturer, is because another } user thinks that this is normal?!
This is EXACTLY THE SORT OF MATTER TO PUT ON THE LISTSERVER.
Even if the boat was not fitting properly, } should the vessel still leak out the front? I would appreciate any } enlightment on this problem. It is a brandnew CPD and thus I have my } doubts that the seal would be damaged already.
We have a Polaron CPD 3000 and the seals often leak, particularly the one on the window. I have taken the CPD apart myself to check it. The window seal leaks even though there are no physical defects on it. And you can replace the same seal in the window and next time it will not leak, indicating no permanent damage. So it is not damaged in the sense that a badly loaded boat has pushed up against it and nicked it so it leaks. In fact I can't see how loading the boat would ever make a difference to the integrity of either of the seals, considering how they are located completely inside grooves.
BUT the door seal can be physically damaged by hard particles (say bits of cover slip) getting washed out of the chamber and locating at the sealing surface so they nick the seal as you screw up the door. SO you need to go carefully around the DOOR sealing surface and screw thread with say ethanol on a cotton bud once a week.
AND the seals on the drain valve on the bottom need regular cleaning for the same reason. Grit washes down the drain and scores the sealing surface and O rings in the valve. In fact if you have never done it, go and clean the door seal and drain valve right away. You'll be surprised at the gunge that will be there.
My explanation is to do with seal elasticity. When the CPD is cooled before being pressurised, the neoprene seals lose much of their elasticity and do not expand properly to seal when the chamber is pressurised. So the solvent leaks out and as the seal stays cold it will never seal properly as of course you keep the chamber cold to ensure fast fluid transfer.
The remedy according to Me is to pressurise the CPD BEFORE COOLING IT. BTW we heat and cool the CPD by circulating water through the jacket using a waterbath heater/mixer (Lauda type T, -20deg to 100 deg C.) which has a small centrifugal pump on it and which sits in a 2 Litre stainless tank. At the start we fill up the small tank with ice and add enough water to cover the heater/pump on the mixer. Then we start the pump with the heater off and chill the CPD that way. When we want to warm the CPD we toss away the ice water, replace it with tap water -at- 20 deg or so and turn on the heater which warms the waterbath towards 50 deg.
Notwithstanding all of the above, since the CPD only works if all the seals are good, you must keep a couple of seal kits in your lab ready for quick repairs. Failure of the seals is usually discovered after some poor soul has spent weeks on their experiment and days on processing their tissue only to find the CPD leaks. If you have the capacity to quickly replace any of the seals so their experiment survives your reputation will be much enhanced!
Mel Dickson E.M. Unit, University of NSW Sydney, Australia
--Boundary (ID uHV94w9QrtOCr/FV43L4kg) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHA38D08MO002DD2-at-mc.duke.edu} ; Thu, 03 Apr 1997 19:23:42 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id TAA03810; Thu, 03 Apr 1997 19:20:34 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id RAA09492 for dist-Microscopy; Thu, 03 Apr 1997 17:26:25 -0600 Received: from sam.comms.unsw.EDU.AU (sam.comms.unsw.EDU.AU [149.171.96.20]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id RAA09489 for {microscopy-at-sparc5.microscopy.com} ; Thu, 03 Apr 1997 17:26:22 -0600 Received: from the_boss (emunit2.med.unsw.EDU.AU [149.171.168.123]) by sam.comms.unsw.EDU.AU (8.7.5/8.7.5.kenso-central) with SMTP id JAA27392 for {microscopy-at-sparc5.microscopy.com} ; Fri, 04 Apr 1997 09:37:48 +1000 (EST)
--Boundary (ID aTDNEG7yxrf2MK3jlfnkUw) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } Can anyone please comment on the storage properties of common EM/LM fixatives. } We have some older 1% glut/4% formaldehyde in PO4 buffer. Is there some rule } of thumb that people are using, should we analyze before use, are there some } references we can access? } Thank you in advance for your comments. } } Regards, } Ken Baker } } The aldehydes oxidise to acids - formic or glutaric. The reaction is less at lower pH so is accelerated when you buffer the solution to pH7. Our rule is to buffer the fixative just before we use it and discard any buffered fixative older than a week.
Mel Dickson }
--Boundary (ID aTDNEG7yxrf2MK3jlfnkUw) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHA3KBIII8003CCZ-at-mc.duke.edu} ; Thu, 03 Apr 1997 19:33:19 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id TAA04007; Thu, 03 Apr 1997 19:30:16 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id RAA09581 for dist-Microscopy; Thu, 03 Apr 1997 17:46:34 -0600 Received: from sam.comms.unsw.EDU.AU (sam.comms.unsw.EDU.AU [149.171.96.20]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id RAA09578 for {microscopy-at-sparc5.microscopy.com} ; Thu, 03 Apr 1997 17:46:31 -0600 Received: from the_boss (emunit2.med.unsw.EDU.AU [149.171.168.123]) by sam.comms.unsw.EDU.AU (8.7.5/8.7.5.kenso-central) with SMTP id JAA08217 for {microscopy-at-sparc5.microscopy.com} ; Fri, 04 Apr 1997 09:58:02 +1000 (EST)
--Boundary (ID 7odngDaNS5WX72Ua9EYGSA) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} We solicit your help in: } (1) Sharing knowledge about similar situations
I used to have a lab 50 metres from a (not so busy) railway line. We could detect the vibration from the trains when they were around a kilometre off.
The problem will vary according to the soil type between your lab and the new road. Ideally the road would be on swampy ground and your lab on a platfrom carved out of granite. That would give good decoupling. But basically, any effective solution in you laboratory will cost several thousands per instrument, as each instrument will need to be relocated on some heavy support (thick steel plate, thicker concrete slab, mass is what you need) with some very flexible mounts under it (air-springs are ideal). It may be more cost effective to route the road further away.
} (2) Pointing us to the best published sources about EM lab design, } particularly in regard to vibration and preferred distance from nearby roads
} (3) Pointing us to any expert EM lab design firms from whom we } might get information
The classic reference work is "Design of the Electron Microscope Laboratory" by Ronald H Alderson 1975. It is Volume 4 in "Practical Methods in Electron Microscopy" Editor Audrey M Glauert. American Elsevier ISBN 0444 1087 6. Was still available two years back whem I bought my second copy to share with the architect for my new laboratory. Pages 68-86 deal with mechanical vibration and decoupling/damping systems } } Mel Dickson } } } }
--Boundary (ID 7odngDaNS5WX72Ua9EYGSA) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHA3MU5RCW003010-at-mc.duke.edu} ; Thu, 03 Apr 1997 19:35:25 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id TAA04031; Thu, 03 Apr 1997 19:32:18 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id RAA09556 for dist-Microscopy; Thu, 03 Apr 1997 17:41:57 -0600 Received: from sam.comms.unsw.EDU.AU (sam.comms.unsw.EDU.AU [149.171.96.20]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id RAA09553 for {microscopy-at-sparc5.microscopy.com} ; Thu, 03 Apr 1997 17:41:54 -0600 Received: from the_boss (emunit2.med.unsw.EDU.AU [149.171.168.123]) by sam.comms.unsw.EDU.AU (8.7.5/8.7.5.kenso-central) with SMTP id JAA05587 for {microscopy-at-sparc5.microscopy.com} ; Fri, 04 Apr 1997 09:53:23 +1000 (EST)
--Boundary (ID VJCFTx1Tb7k8shMYnw8Rcg) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
A Lab6 cathode running on a Hitachi H7100 TEM (manufacturers brand) developed a bright rectanglar-shaped emmision pattern over the last 50-100 hours of operation, although it can be biased back to a small circular spot. We hadnt seen this before. The tip of the cathode shows a distinctly bar-shaped tip when viewed in a light microscope.
The cathode has been very stable over its 450 hours of life, although it seems to move a little vertically as it heats up. There is plenty of Lab6 left.
Does anyone know what causes the pattern, and what are the chances of getting back to a squarish/maltese cross type image if we, say, run it a bit hot for a while?
regards, Sally ---------------------------------------------------------------------- Sally Stowe |Email: stowe-at-rsbs.anu.edu.au Facility Coordinator |Post: ANU Electron Microscopy Unit |ANUEMU (RSBS) Ph 61 6 249 2743 |Australian National Univ. FAX 61 6 249 4891 |Canberra, http://online.anu.edu.au/EMU/home.htm |AUSTRALIA 0200
--Boundary (ID VJCFTx1Tb7k8shMYnw8Rcg) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHAF265CC000380I-at-mc.duke.edu} ; Fri, 04 Apr 1997 01:02:16 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id AAA09350; Fri, 04 Apr 1997 00:59:30 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id WAA10667 for dist-Microscopy; Thu, 03 Apr 1997 22:34:43 -0600 Received: from anugpo.anu.edu.au (anugpo.anu.edu.au [150.203.2.6]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id WAA10664 for {microscopy-at-sparc5.microscopy.com} ; Thu, 03 Apr 1997 22:34:40 -0600 Received: from rsbs-central.anu.edu.au (rsbs-central.anu.edu.au [150.203.38.57]) by anugpo.anu.edu.au (8.8.5/8.8.5) with ESMTP id OAA19748 for {microscopy-at-MSA.microscopy.com} ; Fri, 04 Apr 1997 14:46:12 +1000 (EST) Received: from RSBS-CENTRAL/MAILIN by rsbs-central.anu.edu.au (Mercury 1.21) ; Fri, 04 Apr 1997 14:50:02 +0000 (GMT) Received: from MAILIN by RSBS-CENTRAL (Mercury 1.21); Fri, 04 Apr 1997 14:49:58 +0000 (GMT)
--Boundary (ID VJCFTx1Tb7k8shMYnw8Rcg) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
A Lab6 cathode running on a Hitachi H7100 TEM (manufacturers brand) developed a bright rectanglar-shaped emmision pattern over the last 50-100 hours of operation, although it can be biased back to a small circular spot. We hadnt seen this before. The tip of the cathode shows a distinctly bar-shaped tip when viewed in a light microscope.
The cathode has been very stable over its 450 hours of life, although it seems to move a little vertically as it heats up. There is plenty of Lab6 left.
Does anyone know what causes the pattern, and what are the chances of getting back to a squarish/maltese cross type image if we, say, run it a bit hot for a while?
regards, Sally ---------------------------------------------------------------------- Sally Stowe |Email: stowe-at-rsbs.anu.edu.au Facility Coordinator |Post: ANU Electron Microscopy Unit |ANUEMU (RSBS) Ph 61 6 249 2743 |Australian National Univ. FAX 61 6 249 4891 |Canberra, http://online.anu.edu.au/EMU/home.htm |AUSTRALIA 0200
--Boundary (ID VJCFTx1Tb7k8shMYnw8Rcg) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHAF265CC000380I-at-mc.duke.edu} ; Fri, 04 Apr 1997 01:02:16 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id AAA09350; Fri, 04 Apr 1997 00:59:30 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id WAA10667 for dist-Microscopy; Thu, 03 Apr 1997 22:34:43 -0600 Received: from anugpo.anu.edu.au (anugpo.anu.edu.au [150.203.2.6]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id WAA10664 for {microscopy-at-sparc5.microscopy.com} ; Thu, 03 Apr 1997 22:34:40 -0600 Received: from rsbs-central.anu.edu.au (rsbs-central.anu.edu.au [150.203.38.57]) by anugpo.anu.edu.au (8.8.5/8.8.5) with ESMTP id OAA19748 for {microscopy-at-MSA.microscopy.com} ; Fri, 04 Apr 1997 14:46:12 +1000 (EST) Received: from RSBS-CENTRAL/MAILIN by rsbs-central.anu.edu.au (Mercury 1.21) ; Fri, 04 Apr 1997 14:50:02 +0000 (GMT) Received: from MAILIN by RSBS-CENTRAL (Mercury 1.21); Fri, 04 Apr 1997 14:49:58 +0000 (GMT)
--Boundary (ID jVOfA7APgs//CIJwNU7Htw) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
... snips } user thinks that this is normal?! Even if the boat was not fitting properly, } should the vessel still leak out the front? I would appreciate any } enlightment on this problem. It is a brandnew CPD and thus I have my } doubts that the seal would be damaged already. Perhaps it is just not } seated properly. } } P.S. If anyone has recently purchased a Polaron CPD and finds out that } the seal inside the chamber door keeps falling out, a piece of teflon tape } around the seal works wonders! } } } Susan } } } Susan Carbyn } Atlantic Food and Horticulture Research Centre } Agriculture and Agri-Food Canada } Kentville, Nova Scotia B4N 1J5 } Canada } } Phone: (902) 679-5566 } Fax: (902) 679-2311 } E-mail: carbyns-at-em.agr.ca
Well, I wouldn't consider leaking from the from window normal - get the supplier/manufacturer to sort it out.
However, it might be a handling problem rather than an equipment fault. Make sure you go through all the steps slowly, as sudden temperature changes in particular could be causing differential expansion.
I'd also be unhappy about the tape on the door seal. Although unlikely to cause problems, there is a remote chance it might. A better solution is to hold the metal seal edge on and give it a tap against a hard surface - the idea is to make it slightly oval, so it grips its seating.
Regards, Larry Stoter
--Boundary (ID jVOfA7APgs//CIJwNU7Htw) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHAKPYJWDC003HAR-at-mc.duke.edu} ; Fri, 04 Apr 1997 03:44:14 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id DAA10792; Fri, 04 Apr 1997 03:41:23 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id BAA11075 for dist-Microscopy; Fri, 04 Apr 1997 01:17:00 -0600 Received: from relay-7.mail.demon.net (relay-7.mail.demon.net [194.217.242.9]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id BAA11072 for {Microscopy-at-sparc5.microscopy.com} ; Fri, 04 Apr 1997 01:16:58 -0600 Received: from teknesis.demon.co.uk ([158.152.199.245]) by relay-5.mail.demon.net id aa0528981; Fri, 04 Apr 1997 08:19 -0300 (BST)
--Boundary (ID e4e3yYxyfnkXF3KLwMH7bA) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello Mel and others intetested
Many years ago JEOL News published an article on the design of the EM rooms at the John Innes Institute in the UK. As far as I can recall this dealt in some detail with vibration transmission. We based the design of our EM rooms on this and we have had no vibration problems.
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa eurc-at-giraffe.ru.ac.za - tel: +27 461 318168 - fax: +27 461 24377
--Boundary (ID e4e3yYxyfnkXF3KLwMH7bA) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHAMYPQXZ4002EZV-at-mc.duke.edu} ; Fri, 04 Apr 1997 04:48:33 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id EAA11249; Fri, 04 Apr 1997 04:45:45 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id CAA11216 for dist-Microscopy; Fri, 04 Apr 1997 02:37:25 -0600 Received: from quagga ([146.231.128.2]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id CAA11213 for {microscopy-at-sparc5.microscopy.com} ; Fri, 04 Apr 1997 02:37:11 -0600 Received: from giraffe.ru.ac.za [146.231.128.10] by quagga.ru.ac.za with esmtp (Exim 1.58 #1) id 0wD3Gj-0004tK-00; Fri, 04 Apr 1997 09:18:41 +0200 Received: from GIRAFFE/MERCQ by giraffe.ru.ac.za (Mercury 1.21); Fri, 04 Apr 1997 09:18:36 +0200 Received: from MERCQ by GIRAFFE (Mercury 1.21); Fri, 04 Apr 1997 08:25:02 +0200
--Boundary (ID DHcszqob/V35GKwCaSfY4w) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
A Lab6 cathode running on a Hitachi H7100 TEM (manufacturers brand) developed a bright rectanglar-shaped emmision pattern over the last 50-100 hours of operation, although it can be biased back to a small circular spot. We hadnt seen this before. The tip of the cathode shows a distinctly bar-shaped tip when viewed in a light microscope.
The cathode has been very stable over its 450 hours of life, although it seems to move a little vertically as it heats up. There is plenty of Lab6 left.
Does anyone know what causes the pattern, and what are the chances of getting back to a squarish/maltese cross type image if we, say, run it a bit hot for a while?
regards, Sally ---------------------------------------------------------------------- Sally Stowe |Email: stowe-at-rsbs.anu.edu.au Facility Coordinator |Post: ANU Electron Microscopy Unit |ANUEMU (RSBS) Ph 61 6 249 2743 |Australian National Univ. FAX 61 6 249 4891 |Canberra, http://online.anu.edu.au/EMU/home.htm |AUSTRALIA 0200
--Boundary (ID DHcszqob/V35GKwCaSfY4w) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHAF265CC000380I-at-mc.duke.edu} ; Fri, 04 Apr 1997 01:02:16 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id AAA09350; Fri, 04 Apr 1997 00:59:30 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id WAA10667 for dist-Microscopy; Thu, 03 Apr 1997 22:34:43 -0600 Received: from anugpo.anu.edu.au (anugpo.anu.edu.au [150.203.2.6]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id WAA10664 for {microscopy-at-sparc5.microscopy.com} ; Thu, 03 Apr 1997 22:34:40 -0600 Received: from rsbs-central.anu.edu.au (rsbs-central.anu.edu.au [150.203.38.57]) by anugpo.anu.edu.au (8.8.5/8.8.5) with ESMTP id OAA19748 for {microscopy-at-MSA.microscopy.com} ; Fri, 04 Apr 1997 14:46:12 +1000 (EST) Received: from RSBS-CENTRAL/MAILIN by rsbs-central.anu.edu.au (Mercury 1.21) ; Fri, 04 Apr 1997 14:50:02 +0000 (GMT) Received: from MAILIN by RSBS-CENTRAL (Mercury 1.21); Fri, 04 Apr 1997 14:49:58 +0000 (GMT)
--Boundary (ID vIJCtg1KZTjyZgPIZ0MyCQ) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello Susan
Our Polaron E3000 CPD was bought in 1976 and is still going strong, wuite a lot of use too!
When you say Teflon tape around the seal, I in=magine you mean peripherally rather than through the hole in the middle?!
My experience is that the Doughty/Dowty? seal, the main metal ring plus nitrile (?), tends to split regularly with acetone, every 2-4 months or so. If everything has worked once, then assembly should be ok, it is possible for things to loosen but in my experience that is very unusual. You need to look at the inner face of the seal for any imperfection. I don't know if you get any specail tool, but I made up a steel oblong gizmo which is like a big screwdriver blade whichand gets turned gently with an adjustable wrench.
Hey! I've just realiased, we should have another party for its 21st birthday!!! We get a lot of parties around here, folks!
Best wishes - Keith Ryan Plymouth Marine Lab. UK
--Boundary (ID vIJCtg1KZTjyZgPIZ0MyCQ) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHAOG9OHF40029NX-at-mc.duke.edu} ; Fri, 04 Apr 1997 05:31:00 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id FAA11456; Fri, 04 Apr 1997 05:28:08 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id CAA11150 for dist-Microscopy; Fri, 04 Apr 1997 02:13:18 -0600 Received: from mail.nerc-swindon.ac.uk ([192.171.191.9]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id CAA11147 for {Microscopy-at-Sparc5.Microscopy.Com} ; Fri, 04 Apr 1997 02:13:12 -0600 Received: from kwpf11.nerc-keyworth.ac.uk by mail.nerc-swindon.ac.uk with SMTP (PP) id {02484-0-at-mail.nerc-swindon.ac.uk} ; Fri, 04 Apr 1997 09:25:30 +0100 Received: from Keyworth-Message_Server by wpo.nerc.ac.uk with Novell_GroupWise; Fri, 04 Apr 1997 08:22:16 +0000
--Boundary (ID GIk2ogqCNvwKU7/GA9WMdQ) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Forwarded message:
--Boundary (ID JhCrkojQWqt6agJZ3pVdkw) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thanks to all those who responded in tracing Edington's Practical electron microscopy book. Tech Books distributes it through their distributor "Ceramic Book and literature Service (CBLS)."
CBLS c an be contacted at 703-758-2539 (ph#)
703-758-1518 (fax#)
or
614-374-9458 (ph#) 614-374-8029 (fax#).
Mohan Kalyanaraman
--Boundary (ID JhCrkojQWqt6agJZ3pVdkw) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHA81UVWTS0038EC-at-mc.duke.edu} ; Thu, 03 Apr 1997 21:41:11 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id VAA06190; Thu, 03 Apr 1997 21:38:23 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id TAA10213 for dist-Microscopy; Thu, 03 Apr 1997 19:37:02 -0600 Received: from VAXC.PAU.MOBIL.COM (pau.mobil.com [131.126.24.52]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id TAA10210 for {microscopy-at-sparc5.microscopy.com} ; Thu, 03 Apr 1997 19:36:58 -0600 Received: by vaxc.pau.mobil.com (UCX V4.0-10B, OpenVMS V5.5-2 VAX); Thu, 03 Apr 1997 20:46:41 -0500 Received: from ccMail by ccmail.pau.mobil.com (SMTPLINK V2.11.01) id AA860129221; Thu, 03 Apr 1997 20:41:16 -0500 (EST)
-- [ From: Blackwood, Andy * EMC.Ver #3.0 ] --
3 April 1997
Greetings, John Bozzola:
I saw your posting about the JEOL sample a few days ago, and since nobody else seems to have made the point, I thought that a few comments might help. My bias is rather obvious--as Laboratory Director of Structure Probe, Inc., I am responsible for the production of the SPI Supplies reference samples.
We offer three different samples, because our customers request various sizes. All are the same gold on carbon construction, and they offer sharp contrast between gold (high emission of secondary electrons) and carbon (low emission of secondary electrons). We characterize the samples by the average size of the particles (small is around 10 nm, medium is around 30 nm and large is around 100 nm). The actual reference, however, is the gap between gold particles, and by looking around a little, you can find any size gap you wish to use on any of the samples. Larger laboratories have obtained a set of three samples to cover the entire range. Details may be found on our web site.
It certainly is possible to set up to produce such samples in your own laboratory, but by the time you've finished fine tuning the "art" for your local conditions, and then verified that what you made is what you hoped to make, you may conclude that it would have been easier to obtain a commercially made sample.
Andy
Andrew W. Blackwood, Ph.D. Structure Probe, Inc. P.O. Box 656 West Chester, PA 19381-0656 Ph: 1 610 436 5400 FAX: 1 610 436 5755 e-mail: ablackwood-at-2spi.com WWW: http://www.2spi.com
--Boundary (ID Kw7J5YysFyAAlVTueI7Ikw) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Looking for experienced TEM Service Engineer to service Philips 420 in Kansas City Area. Contact Pete Dondl at sylviapns-at-worldnet.att.net P & S Products, Inc.
--Boundary (ID Kw7J5YysFyAAlVTueI7Ikw) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHAW788T00003AA7-at-mc.duke.edu} ; Fri, 04 Apr 1997 09:12:44 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id JAA16710; Fri, 04 Apr 1997 09:09:38 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id HAA11890 for dist-Microscopy; Fri, 04 Apr 1997 07:10:58 -0600 Received: from mtigwc03.worldnet.att.net (mailhost.worldnet.att.net [204.127.131.34]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id HAA11887 for {MICROSCOPY-at-MSA.MICROSCOPY.COM} ; Fri, 04 Apr 1997 07:10:57 -0600 Received: from default ([207.116.103.162]) by mtigwc03.worldnet.att.net (post.office MTA v2.0 0613 ) with ESMTP id AAA23516 for {MICROSCOPY-at-MSA.MICROSCOPY.COM} ; Fri, 04 Apr 1997 13:21:12 +0000
--Boundary (ID Ztz18ugNGqbMTReFH0PHsw) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
In reply to: ================================================== I used to have a lab 50 metres from a (not so busy) railway line. We could detect the vibration from the trains when they were around a kilometre off. ================================================== There have been instances where the problem with the rail line has been less due to ground vibrations than to the creation of a transient magnetic field problem that correlated with the passage of a train. In about 1969, there was an SEM installed at a Philadelphia university near the main passenger line of AMRAK and Conrail, and the real problem was more related to the magnetic field (trains were pantograph (electrically) powered) than to vibrations. The problem was ultimately "solved" only by moving the microscope. So don't forget the magnetic field problem potential as well.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
--Boundary (ID Ztz18ugNGqbMTReFH0PHsw) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHAWZM454W003DX7-at-mc.duke.edu} ; Fri, 04 Apr 1997 09:35:39 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id JAA17314; Fri, 04 Apr 1997 09:32:20 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id HAA11931 for dist-Microscopy; Fri, 04 Apr 1997 07:25:05 -0600 Received: from ns1.axs2000.net (ns1.axs2000.net [207.106.10.100]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id HAA11928 for {Microscopy-at-sparc5.microscopy.com} ; Fri, 04 Apr 1997 07:25:04 -0600 Received: from [205.186.65.46] (tfx-us7-14.ix.netcom.com [205.186.65.46]) by ns1.axs2000.net (8.6.13/8.6.12) with SMTP id IAA05363 for {Microscopy-at-MSA.Microscopy.com} ; Fri, 04 Apr 1997 08:15:43 -0500
--Boundary (ID 50CJZhSiyB9iKyfaPPasDA) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi Candace. There was a discussion a while back on this topic which has been archived at the "Tips & Tricks" site. Go to the web address listed at the end of this message and follow the "Tips & Tricks link. Proceed to the Miscl. link and there it is at the top of the list. Good luck.
At 10:07 AM 4/3/97 +0131, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 352-392-1295 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
--Boundary (ID 50CJZhSiyB9iKyfaPPasDA) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHAXINOIKW002ATT-at-mc.duke.edu} ; Fri, 04 Apr 1997 09:50:57 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id JAA17862; Fri, 04 Apr 1997 09:48:05 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id HAA11873 for dist-Microscopy; Fri, 04 Apr 1997 07:06:42 -0600 Received: from snarl.biotech.ufl.edu (snarl.biotech.ufl.edu [128.227.186.119]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id HAA11870 for {microscopy-at-Sparc5.Microscopy.Com} ; Fri, 04 Apr 1997 07:06:41 -0600 Received: from scope.biotech.ufl.edu by snarl.biotech.ufl.edu (8.7/4.09) id IAA25755; Fri, 04 Apr 1997 08:12:52 -0500 (EST)
On Fri, 4 Apr 1997 sylviapns-at-worldnet.att.net wrote:
} Date: Fri, 04 Apr 1997 11:12 -0500 (EST) } From: sylviapns-at-worldnet.att.net } To: MICROSCOPY-at-Sparc5.Microscopy.Com } Subject: TEM Service } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Looking for experienced TEM Service Engineer to service Philips 420 in } Kansas City Area. } Contact } Pete Dondl at sylviapns-at-worldnet.att.net } P & S Products, Inc. } Call Philips: 1 800 432 1PEI}
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Another interesting problem I was told about related to a EM installed in Eastern Europe a number of years back. There were continual, intermittent resolution and noise problems, worse during the day, not too often at night and after midnight, the problems disappeared.
After a lot of investigations, the problems was finally traced to the local trams. What was happening was that everything was OK until the trams reached a nearby hill - the extra load in pulling up the hill (trams going down the hill weren't a problem) dragged the mains supply voltage below an acceptable level and caused a whole series of instabilities in the EM.
Regards,
--------------------------------------------------------------- Dr L. P. Stoter Technical Editor, MICROSCOPY & ANALYSIS Technesis 17, Rocks Park Road email: LPS-at-teknesis.demon.co.uk Uckfield, E. Sussex Phone: +44 (0)1825 766911 TN22 2AT Fax: +44 (0)1825 766911 United Kingdom ---------------------------------------------------------------
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Postdoctoral Positions in Biophysics, Biochemistry and Biomathematics.
Two NSF-funded fellowships are available immediately for interdisciplinary studies of macromolecular assemblies in the following areas:
1. Computational analysis of atomic interactions: including development of holistic mathematical analysis of fundamental interactions, exploiting the complementarity of data from diffraction, NMR spectroscopy and electron microscopy. Candidates should have backgrounds in computational methods development in crystallography, NMR, EM or computational structural biology, or training in the mathematical / physical sciences combined with demonstrable interest in structural biology.
2. Structure & assembly of macromolecular complexes of fibroblast growth factor & receptor: the candidate will pursue structural analyses of soluble forms of fibroblast growth factor receptors and receptor/growth factor complexes. Candidates should have a background in x-ray crystallography, protein purification and molecular biology. Experience in baculovirus expression systems, and cell culture would be a plus.
3. Comparing protein motion in solution and the crystal. The fellow will develop an approach to completely map the dynamics of the C-terminal domain of the diptheria toxin repressor protein by combining solution NMR spectroscopy and diffuse-scatter X-ray crystallography. Candidates should have a strong background in NMR spectroscopy or x-ray crystallography, with an interest in protein dynamics.
4. Protein dynamics as measured by EPR and X-ray diffraction methods: the candidate will develop "rules" governing the dynamics of spin labels attached to proteins of known structure. The dynamics observed in the spin labeled crystals using EPR will be compared to the diffuse scatter and the temperature factors from x-ray studies. The candidate should have background in magnetic resonance techniques or x-ray crystallography.
Consistent with these interdisciplinary fellowships, trainees will be mentored jointly by faculty from at least two fields: Michael Blaber, Don Caspar and Michael Chapman (crystallography), Tim Cross and Tim Logan (NMR), Piotr Fajer (EPR) and Ken Taylor (EM). Supplementing experimental facilities at the Institute of Molecular Biophysics, close ties are enjoyed with the National High Field Magnetic Laboratory and the Supercomputer Computation Research Institute on campus. Fellows will enjoy a stimulating intellectual environment, pleasant weather, national forests and pristine gulf beaches.
Candidates should send a resume and arrange for 3 letters of reference to be forwarded to the Institute of Molecular Biophysics, Attn: Carolyn Moore / Post-doc. Fellows, Florida State University, Tallahassee, FL 32306-3015, USA, prior to July 1st. The project of interest should be stated at the top of the resume. Additional information about the Structural Biology Program can be found at http://www.sb.fsu.edu/.
-- Michael S. Chapman (chapman-at-sb.fsu.edu) http://www.sb.fsu.edu/~chapman/ Assistant Prof. Chemistry (904) 644-8354 FAX: (904) 644-3257 Institute of Molecular Biophysics, Florida State University, Tallahassee, FL 32306-3015
Speaking of vibrations, can anyone provide information (name, address, phone, FAX, e-mail, www address, etc.) for vendors of vibration isolation pads and platforms. I have a new stereo microscope installation that is being bothered by building vibrations.
Thanks...
Larry Sutter Michigan Technological University Dept. of Civil and Environmental Engineering 1400 Townsend Dr. Houghton, MI 49931
At 06:30 PM 06-04-97 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
You can do it yourself.
Get a 2 ft (or so) square of paving cement from a garden supply (or hardware ) company. Buy 4 tennis balls. Put the slab on some sturdy bench with a tennis ball under each corner. Put the microscope on the slab. For a more compliant isolator use a small inner tube from some small wheel.
The problem does not appear to originate at the Microscopy Listserver. It looks to be a computer/email system at DUKE.EDU that is redirecting mail back to the server. For the moment I have removed the following addresses from the subscription list. If this cures the problem we know that the problem has been at least isolated.
One of my colleagues asks: } } Does anyone know where I can get a program that can calculate } the multi-beam diffraction contrast of a (given) strain field? } (i.e. not just a two-beam calculation)
TIA,
Stephen. ............................................................... : Stephen Anderson Australian Key Centre for : : Microscopy and Microanalysis : : Email stephen-at-emu.usyd.edu.au The University of Sydney : : Telephone (+61)-2-9351 7552 NSW 2006 : : Facsimile (+61)-2-9351 7682 Australia : :.............................................................:
We sometimes store small aliquots of fixative at minus 20 C. I'm not sure this is effective but it seems like a reasonable thing to do. Seems as though I came upon this in one of the early editions of Hyats general EM techniques book. ******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Colleagues...
The Microscopy Node was down for most of the day today. There was another problem with Ameritek. It must be all that warm weather we are having in Chicago ;-)
As for the problem about the duplicate postings as far as I can tell the problem does not appear to originate at the Microscopy Listserver. It looks to be a computer/email system at DUKE.EDU that is redirecting mail back to the server. For the moment I have removed the following addresses from the subscription list. If this cures the problem we know that the problem has been at least isolated.
there is a program called SIMCON that will probably do what you want. It's written at the materials department of the University of Leuven by Koen Janssens. You'll find a reference for it in "Ultramicroscopy 45, p. 323 (1992)" and at "http://www.mtm.kuleuven.ac.be/~simcon/". The author has now left the group and works at OCAS, J.F. Kennedylaan 3, B-9060 Zelzate.
1. When a gold coated specimen is viewed in SEM at ~10k or higher, a fine structure of gold coating becomes visible. Are there optimal conditions of sputter coating (sputter current, time, distance target-specimen, argon pressure, other gas than argon) to minimise the artefact?
2. What is Au-Pd target? I thought that it was just an alloy one, but a supplier of the coater says that using the alloy target is not enough, that a coater with simultaneous sputtering Au and Pd from two different targets is required to produce continuous coating.
Of course, it's better to get a FEG and use low voltage, but I want to do it with a magnetron sputter coater and conventional JSM 6400.
Thank you,
Alex
__________________ Alexander Titkov
Millennium Inorganic Chemicals PO Box 245 Bunbury WA 6231 Australia Ph (097) 808 505 FAX: (097) 808 444 E-mail: scm!atitkov-at-scmaust.attmail.com
This last piece of information was passed along to me by the Manufacturer of the Polaron CPD.
Please ammend the e-mail or notify the customer with the faulty CPD that the seals used were not in fact made by DOWTY but of similar design, from another manufacturer, these in fact were of the wrong material so in practice the material was not faulty but the manufactured item itself was below spec.
Susan
Susan Carbyn Atlantic Food and Horticulture Research Station Agriculture and Agri-Food Canada Kentville, Nova Scotia B4N 1J5 Canada
Meeting: Spring Meeting of the Appalachian REgional Microscopy Society Dates: April 17 & 18, 1997 Topic: Registration at Comfort Suites, Noon to 5pm, Thursday, 4/17/97 Workshops 1pm to 5pm, Thursday, 4/17/97 Social and Banquet, 6pm, Thursday, 4/17/97 Technical Presentations at Enka Lake Club, 8am to 1pm, Friday, 4/18/97 Sponsor: BASF, Enka, NC Location: Asheville, NC, USA Interests: Both Physical & Biological Sciences Fields: Light/Optical, SEM, TEM Contact: Susan Read, BASF Corporation, Sand Hill Road, Enka, NC, 28728, USA Tel: (704)667-6353 Fax: (704)667-6903 E-mail: reads-at-basf.com WWW: http://www.clinck.edu/~jrb/arems.html ---------------------------------------------------------------------------
--Boundary (ID pZyvR9B+N6pmEwGN7oHkGg) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Susan,
} } We have recently purchased a Polaron CPD and wanted to hear from } anyone else who has one. I had used one at a University, at which time } once I loaded the specimen chamber and shut the back, opened the fill } valve, the CO2 leaked out the front window. I was concerned and drained } out the CO2. I went to find the instructor, and asked why this was } happening. I assumed a seal was faulty, but she told me that the boat } wasn't loaded properly. The back had closed nicely, and I really didn't } think that it was not loaded properly. Once the chamber was reloaded, } the CO2 tank was empty and I could not finish the run. Once the CO2 } tank was replaced weeks later, I received a phone call saying that the seal } was damaged and that I would have to wait before the new ones came in. } To make a long story short, we now have a Polaron CPD and I am having } the same problem. Sometimes when I load it, it leaks out the front. } Everything seems to be fitting properly, but on occasion, upon filling, it } leaks out the front window. The only reason that I am posting this } question to the listserver and not to the manufacturer, is because another } user thinks that this is normal?!
This is EXACTLY THE SORT OF MATTER TO PUT ON THE LISTSERVER.
Even if the boat was not fitting properly, } should the vessel still leak out the front? I would appreciate any } enlightment on this problem. It is a brandnew CPD and thus I have my } doubts that the seal would be damaged already.
We have a Polaron CPD 3000 and the seals often leak, particularly the one on the window. I have taken the CPD apart myself to check it. The window seal leaks even though there are no physical defects on it. And you can replace the same seal in the window and next time it will not leak, indicating no permanent damage. So it is not damaged in the sense that a badly loaded boat has pushed up against it and nicked it so it leaks. In fact I can't see how loading the boat would ever make a difference to the integrity of either of the seals, considering how they are located completely inside grooves.
BUT the door seal can be physically damaged by hard particles (say bits of cover slip) getting washed out of the chamber and locating at the sealing surface so they nick the seal as you screw up the door. SO you need to go carefully around the DOOR sealing surface and screw thread with say ethanol on a cotton bud once a week.
AND the seals on the drain valve on the bottom need regular cleaning for the same reason. Grit washes down the drain and scores the sealing surface and O rings in the valve. In fact if you have never done it, go and clean the door seal and drain valve right away. You'll be surprised at the gunge that will be there.
My explanation is to do with seal elasticity. When the CPD is cooled before being pressurised, the neoprene seals lose much of their elasticity and do not expand properly to seal when the chamber is pressurised. So the solvent leaks out and as the seal stays cold it will never seal properly as of course you keep the chamber cold to ensure fast fluid transfer.
The remedy according to Me is to pressurise the CPD BEFORE COOLING IT. BTW we heat and cool the CPD by circulating water through the jacket using a waterbath heater/mixer (Lauda type T, -20deg to 100 deg C.) which has a small centrifugal pump on it and which sits in a 2 Litre stainless tank. At the start we fill up the small tank with ice and add enough water to cover the heater/pump on the mixer. Then we start the pump with the heater off and chill the CPD that way. When we want to warm the CPD we toss away the ice water, replace it with tap water -at- 20 deg or so and turn on the heater which warms the waterbath towards 50 deg.
Notwithstanding all of the above, since the CPD only works if all the seals are good, you must keep a couple of seal kits in your lab ready for quick repairs. Failure of the seals is usually discovered after some poor soul has spent weeks on their experiment and days on processing their tissue only to find the CPD leaks. If you have the capacity to quickly replace any of the seals so their experiment survives your reputation will be much enhanced!
Mel Dickson E.M. Unit, University of NSW Sydney, Australia
--Boundary (ID pZyvR9B+N6pmEwGN7oHkGg) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHA38D08MO002DD2-at-mc.duke.edu} ; Thu, 03 Apr 1997 19:23:42 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id TAA03810; Thu, 03 Apr 1997 19:20:34 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id RAA09492 for dist-Microscopy; Thu, 03 Apr 1997 17:26:25 -0600 Received: from sam.comms.unsw.EDU.AU (sam.comms.unsw.EDU.AU [149.171.96.20]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id RAA09489 for {microscopy-at-sparc5.microscopy.com} ; Thu, 03 Apr 1997 17:26:22 -0600 Received: from the_boss (emunit2.med.unsw.EDU.AU [149.171.168.123]) by sam.comms.unsw.EDU.AU (8.7.5/8.7.5.kenso-central) with SMTP id JAA27392 for {microscopy-at-sparc5.microscopy.com} ; Fri, 04 Apr 1997 09:37:48 +1000 (EST)
} ------------------------------------------------------------------------ Alex wrote, } 1. When a gold coated specimen is viewed in SEM at ~10k or higher, a } fine structure of gold coating becomes visible. Are there optimal } conditions of sputter coating (sputter current, time, distance } target-specimen, argon pressure, other gas than argon) to minimise the } artefact? } } 2. What is Au-Pd target? I thought that it was just an alloy one, but } a supplier of the coater says that using the alloy target is not } enough, that a coater with simultaneous sputtering Au and Pd from two } different targets is required to produce continuous coating.
Dear Alex, 1. Assuming that you are seeing gold, this indicates that your coating is too thick. When a sample is coated it should take on a bluish color. This is most easistly seen on flat areas where there is no sample. Try using a cover slip to set up the conditions. AS to conditions. Don't rush the coating. It should take about a minute to coat your sample. The manuel that came with the sputter coater will give you a starting point.
2. You are right, it is an alloy target. This target should give you a finer grain size, but the secondary electron return is not as good.
Gregory Rudomen Greg-at-UMIC.SUNYSB.EDU 516-444-3126 University Microscopy Imaging Center S.U.N.Y. Stony Brook
Meeting: Tripod Polisher Workshop Dates: June 6 & 7 1997
Topic: This course will cover all aspects of pre-thinning for TEM and will 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 and in Plasma Cleaning for TEM samples.
Sponsor: South Bay Technology, Inc. Location: San Clemente, CA, USA Interests: Physical Sciences Fields: SEM, TEM, STEM, IVEM/HVEM, AEM Contact: Monica Pflaster, South Bay Technology, Inc., 1120 Via Callejon, San Clemente, CA, 92673, USA Tel: 800-728-2233 Fax: 714-492-1499 E-mail: sbt-at-southbaytech.com WWW: http://www.southbaytech.com ---------------------------------------------------------------------------
--Boundary (ID znp7YOOngc6OXbMhtxtdig) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } Can anyone please comment on the storage properties of common EM/LM fixatives. } We have some older 1% glut/4% formaldehyde in PO4 buffer. Is there some rule } of thumb that people are using, should we analyze before use, are there some } references we can access? } Thank you in advance for your comments. } } Regards, } Ken Baker } } The aldehydes oxidise to acids - formic or glutaric. The reaction is less at lower pH so is accelerated when you buffer the solution to pH7. Our rule is to buffer the fixative just before we use it and discard any buffered fixative older than a week.
Mel Dickson }
--Boundary (ID znp7YOOngc6OXbMhtxtdig) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHA3KBIII8003CCZ-at-mc.duke.edu} ; Thu, 03 Apr 1997 19:33:19 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id TAA04007; Thu, 03 Apr 1997 19:30:16 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id RAA09581 for dist-Microscopy; Thu, 03 Apr 1997 17:46:34 -0600 Received: from sam.comms.unsw.EDU.AU (sam.comms.unsw.EDU.AU [149.171.96.20]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id RAA09578 for {microscopy-at-sparc5.microscopy.com} ; Thu, 03 Apr 1997 17:46:31 -0600 Received: from the_boss (emunit2.med.unsw.EDU.AU [149.171.168.123]) by sam.comms.unsw.EDU.AU (8.7.5/8.7.5.kenso-central) with SMTP id JAA08217 for {microscopy-at-sparc5.microscopy.com} ; Fri, 04 Apr 1997 09:58:02 +1000 (EST)
} 1. When a gold coated specimen is viewed in SEM at ~10k or higher, a } fine structure of gold coating becomes visible. Are there optimal } conditions of sputter coating (sputter current, time, distance } target-specimen, argon pressure, other gas than argon) to minimise the } artefact?
You will probably need to experiment, since the optimum coating will depend to some extent on your specimen - rough specimens will need a coarser, larger grained coating to prevent charging. In addition, you will get a huge amount of advice from all directions, much of it contradictory! so you'll only find out what is right for you by experiment. Anyway, MY advice is:
Argon is generally the best option and you also want to make sure that the argon isn't contaminated with N or O - this will substantially reduce the sputtering rates. If you're Ar supply is good, then this only requires that you flush the system for a short period before sputtering.
Argon pressure/flow rate should be adjusted so that the plasma is just steady - turn up the Ar flow rate until you get a plasma, and then slowly reduce the flow rate (be slow because there will be a significant lag between changing the flow rate and the system stabilising). At some point, the plasma will start to flicker, and then go out. Increase the flow rate to just higher than the point at which the plasma flickers (obviously, this all needs to be setup either without the specimen in the coater, or with a shutter over the specimen).
Lower voltages and shorter times will tend to produce finer and thinner coatings, respectively. Most suppliers will advise approx 1.5 to 1.8 kV, but you can produce some very nice coatings at 600 to 800 V. You should coat for a period just long enough to stop specimen charging.
} 2. What is Au-Pd target? I thought that it was just an alloy one, but } a supplier of the coater says that using the alloy target is not } enough, that a coater with simultaneous sputtering Au and Pd from two } different targets is required to produce continuous coating.
Au/Pd target is an alloy - I'm not aware of commercial coaters that simultaneously work from a pair of targets. Such an arrangement might be more effective but I want some evidence that it was, and I expect it would be more expensive:) Au/Pd alloy targets are effective at producing finer grained coatings. It seems that the Pd provides nuclei for the Au, leading to more, and smaller Au grains rather than the Au grains growing larger as with a pure Au target.
} Of course, it's better to get a FEG and use low voltage, but I want } to do it with a magnetron sputter coater and conventional JSM 6400. } } Thank you, } } Alex
--Boundary (ID c5OrUMZzajGDuj5qW8trnQ) Content-type: TEXT/PLAIN
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} We solicit your help in: } (1) Sharing knowledge about similar situations
I used to have a lab 50 metres from a (not so busy) railway line. We could detect the vibration from the trains when they were around a kilometre off.
The problem will vary according to the soil type between your lab and the new road. Ideally the road would be on swampy ground and your lab on a platfrom carved out of granite. That would give good decoupling. But basically, any effective solution in you laboratory will cost several thousands per instrument, as each instrument will need to be relocated on some heavy support (thick steel plate, thicker concrete slab, mass is what you need) with some very flexible mounts under it (air-springs are ideal). It may be more cost effective to route the road further away.
} (2) Pointing us to the best published sources about EM lab design, } particularly in regard to vibration and preferred distance from nearby roads
} (3) Pointing us to any expert EM lab design firms from whom we } might get information
The classic reference work is "Design of the Electron Microscope Laboratory" by Ronald H Alderson 1975. It is Volume 4 in "Practical Methods in Electron Microscopy" Editor Audrey M Glauert. American Elsevier ISBN 0444 1087 6. Was still available two years back whem I bought my second copy to share with the architect for my new laboratory. Pages 68-86 deal with mechanical vibration and decoupling/damping systems } } Mel Dickson } } } }
--Boundary (ID c5OrUMZzajGDuj5qW8trnQ) Content-type: TEXT/PLAIN
RFC-822-headers: Received: from nexus.mc.duke.edu (nexus.mc.duke.edu) by mc.duke.edu (PMDF V5.0-5 #16741) id {01IHA3MU5RCW003010-at-mc.duke.edu} ; Thu, 03 Apr 1997 19:35:25 -0500 (EST) Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by nexus.mc.duke.edu (8.7.5/8.7.3) with SMTP id TAA04031; Thu, 03 Apr 1997 19:32:18 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id RAA09556 for dist-Microscopy; Thu, 03 Apr 1997 17:41:57 -0600 Received: from sam.comms.unsw.EDU.AU (sam.comms.unsw.EDU.AU [149.171.96.20]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id RAA09553 for {microscopy-at-sparc5.microscopy.com} ; Thu, 03 Apr 1997 17:41:54 -0600 Received: from the_boss (emunit2.med.unsw.EDU.AU [149.171.168.123]) by sam.comms.unsw.EDU.AU (8.7.5/8.7.5.kenso-central) with SMTP id JAA05587 for {microscopy-at-sparc5.microscopy.com} ; Fri, 04 Apr 1997 09:53:23 +1000 (EST)
Hi I am looking for information about ccd analogous or digital cameras to couple to my optical microscope OLYMPUS BX50. Wanted to know of the fact that resolution must be to use it in reconstruction 3D and digital images procesing. If furthermore know the address e-mail of some providing, them would thank that me facilitate it Thank you very much
Fernando Diego Balducci Laboratory of Electron Microscopy School of Engineery - Bioengineery National University of Entre Rios Argentina Phone: 54 43 975100 Fax : 54 43 975077 e-mail : RNBALDUC-at-ARCRIDE.EDU.AR
1997 ADVANCED SPECIMEN PREPARATION WORKSHOPS (for LM, SEM/TEM, and SPM)
The following 8 independent workshops offer an intensive hands-on training program for the application of the most advanced specimen preparation techniques currently available for microscopy of complex material systems. These workshops are intended for R&D personnel involved in microscopy of advanced materials and/or related specimen preparation. Enrollment is limited to 4 students in each workshop and early registration is strongly recommended to ensure admission.
***Site-specific Cross-sectioning and Microthinning Precision Cleaving (May 6 or Nov. 4 in Santa Clara, CA) FIB-milling for SEM Cross-sectioning (May 7 or Nov.5 in Sunnyvale, CA) FIB-milling for TEM Cross-sectioning (May 7-9 or Nov.5-7 in Sunnycale, CA) Precision Lapping for SEM Cross-sectioning (May 14 or Nov. 12 in Phoenix, AZ) TEM Wedge-polishing (May 14-16 or Nov. 12-14 in Phoenix, AZ)
***Materials Ultramicrotomy General Surface Preparation for LM, SEM, or AFM (May 19-20 or Nov. 17-18 in Phoenix, AZ) Thin Section Preparation for TEM (May 19-21 or Nov. 17-19 in Phoenix, AZ) Advanced Ultramicrotomy (May 22-23 or Nov 20-21 in Phoenix, AZ)
A partial list of the past participants in these workshops include: IBM, Motorola, SEMATECH, Texas Instruments, Medtronic, Hewlett-Packard, Lawrence Berkeley Lab, Oak Ridge National Lab, National Renewable Energy Lab, Bell Northern Research (Canada), Honeywell, Martin Marietta, B.F. Goodrich, 3M, MIT Lincoln Lab, United Technologies, Hydro Quebec (Canada), Cabot, Lawrence Livermore National Lab, US Army Research Lab, Kimberly Clark, etc.
For further information and on-line registration, please see our home page hosted on Microscopy Online at http://www.microscopy-online.com/Vendors/AMCGroup/. You may also request a copy of the workshops brochure by providing us with your complete mailing address.
Rene E. Nicholas AMC Group (a Division of Promotech Associates, Inc.) amcgroup2-at-aol.com
Is your web site the best kept secret on the Internet?
We'll promote it to 50 search engines and indexes for $85 and complete the job in 2 business days. Satisfaction is guaranteed!
If you have a great product, but are not getting many inquiries from your Web site, you may not be adequately listed on the Web's search engines and indexes.
Millions of viewers daily use these facilities to find the products and services they are looking for. But if your site is not listed, no one will see it.
Listings on most of these services are free. However, locating and filling out the forms required to get a listing can take several days, and most people just don't have the time to do it.
That is why we offer a web site promotion service.
WHAT'S THE DEAL?
We will submit your site to 50 indexes and search engines for $85. We will accept the return of this E-mail, with the form below filled out, as an order. We will bill you upon completion of the promotion. Our terms are net 15 days from date of invoice. Satisfaction guaranteed!
HOW LONG WILL IT TAKE?
Generally, we complete the submissions within 48 hours of receiving your order. It can take any individual search engine or index up to three weeks to process your submission, although most are much faster.
WHAT SEARCH ENGINES AND INDEXES ARE INCLUDED IN THE PROMOTION?
The list changes from time to time. This is our current list:
Abaweb!, Alta Vista, Been There, BizWeb, Central Source Yellow Pages, Enterpreneurs on the Web, Excite, Four11, Galaxy, I-Network I-Systems Spiral Business Directory, I-World Web Pointer, Infoseek, Inktomi, Innovator's Network Yellow Pages, Internet Mall, Jayde Online Directory, Jumpcity, Jumper Hot Links, Linkmaster, Lycos, Magellan, Mega Mall, Net-Happenings, Net Navigator, Net Mall, NTG's List, NYNEX Big Yellow, One World Plaza, OnLine's WWWeb Index, Rex, Starting Point, Truenorth, URL Tree, Virtual Lynx, Web Point, WebCentral, Web Venture Hotlist, Webcrawler, Websurf, Win Mag/NetGuide Hotspots, WhatUSeek, Worldwide Announce Archive, WWW Business Yellow Pages, World Wide Yellow Pages, WWW Worm, YelloWWWeb.
HOW WILL I KNOW THAT YOU HAVE PROMOTED MY SITE?
When we have completed the promotion, we will send you an HTML file as an attachment to your E-mail bill. Save this file to your disk, and view it through your Web browser. It provides links to the search engine we submitted your site to, plus any comments we received from them when we did it.
ARE THERE ANY GUARANTEES?
We do not require prepayment. Your satisfaction is guaranteed or you don't pay the bill.
WHO IS OWL'S EYE PRODUCTIONS?
We are a web site promotion company located at:
Owl's Eye Productions, Inc. 260 E. Main Street Brewster, NY 10509 Phone: (914) 278-4933 Fax: (914) 278-4507 Email: owl-at-owlsnest.com
HOW DO I ORDER?
The easiest way to order is by e-mail. Just hit the REPLY button on your e-mail program and fill out the following information. (This information will be posted to the search engines/indexes):
Your name: Company Name: Address: City: State/Prov: Zip/Postal Code: Telephone: Fax: Email address: URL: http:// Site Title: Description (about 25 words): Key words (maximum of 25, in descending order of importance):
Proofs (Where shall we e-mail proofs):
If billing a different address, please complete the following:
Terms: By returning this document via Email, you agree as follows: You have the authority to purchase this service on behalf of your company. Terms are net 15 days. Accounts sent to collections will be liable for collection costs. You agree to protect and indemnify Owl's Eye Productions, Inc. in any claim for libel, copyright violations, plagiarism, or privacy and other suits or claims based on the content or subject matter of your site.
WHAT HAPPENS NEXT?
When we receive your order, we will input the information into our system, and send you a proof.
After we process any corrections, we will run your promotion, capturing any comments from search engines as we go. We will incorporate these into an HTML-formatted report to you, which we will attach to your bill. ===Web Promotions=====Press Releases=====Link Exchanges========= Owl's Eye Productions, Inc. 260 E. Main Street Brewster, NY 10509 Ph: 914-278-4933 Fx: 914-278-4507 E-mail: owlseye-at-owlsnest.com
Dear Alex, You should not see the structure of a Au-Pd film at 10K. In the 1980 SEM Inc. Conference Proceedings there is an extensive study of the various conditions for making films, with the films studied by TEM. It is worth reading if you have access to it. The result was a recommendation to use Ar gas, 60%Au-40%Pd instead of Au and a lower voltage, 600 to 700 volts. The specimen-surface distance is about 2 cm. I have been using a 1 to 3 minute coating in Ar at 700v. ever since and can only see the film at 50,000 times. Coatings using W or Ni were even finer. I have never heard of using two targets instead one alloyed one. I'm sure the ionized particles don't care. The very fine coatings required by FEG microscopes are best made by an ion-beam coater (much more expensive). You wrote: } Two questions: } } 1. When a gold coated specimen is viewed in SEM at ~10k or higher, a } fine structure of gold coating becomes visible. Are there optimal } conditions of sputter coating (sputter current, time, distance } target-specimen, argon pressure, other gas than argon) to minimise the } artefact? } } 2. What is Au-Pd target? I thought that it was just an alloy one, but } a supplier of the coater says that using the alloy target is not } enough, that a coater with simultaneous sputtering Au and Pd from two } different targets is required to produce continuous coating. } } Of course, it's better to get a FEG and use low voltage, but I want } to do it with a magnetron sputter coater and conventional JSM 6400. } } Thank you, } } Alex Good luck, Mary
I bought a gold/palladium coater recently and found that the default time listed in the manual (} 1 minute) resulted in a coating about the thickness of the chrome on a 1950's Buick. I now coat even the most problematic samples (Mo oxide crystals, glass fractures, rare-earth phosphor particles, etc.) for 15 seconds or less. I also dropped the current to about 70% of the spec value. The samples still generate Au and/or Pd x-rays occasionaly, but I still detect light elements. It's a balancing act.
Remember, it is easier to recoat than de-coat.
my 2 cents.
------------------------------------------------ Opinions or statements expressed herein, rational or otherwise, do not necessarily reflect those of my employer.
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 Phone: (508) 750-1717 E-mail: crossman-at-osi.sylvania.com
Our web sites: www.sylvania.com www.siemens.com --
To all on the list I am looking for a lab or person that can do lead phase speciation to distinguish lead products from a smelter from naturally occuring lead minerals. Please reply as soon as possible, there is probably some money and work involved.
Clarissa Vierrether The Doe run Company 573-244-8109 Viburnum, MO
------------------------------------------------------------------------=20 The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20 =20 =20 Alex; =20 In my experience, it would be very unusal to be able to see any detail= s of=20 a Au sputtered coating of normal thickness (5-20 nm.) at a magnificati= on of=20 only 10,000x. Coating artifacts usually become a problem at much high= er=20 magnification such as 50,000x and above. =20 If your sputtered coating is very thick this could be a problem and yo= u=20 should review the operation of your specific coater to determine the b= est=20 parameters for the coating thickness that you desire. The sputter curr= ent,=20 gas pressure, gas quality, distance to the target, and sputtering tim= e are=20 all factors that must be considered. =20 While Au provides an efficient emitter of secondary electrons in the S= EM,=20 the grain size is quite large and does become a problem at higher mags.= =20 Au/Pd is usually an alloy target that combines the better secondary=20 electron emission characteristics of Au along with the smaller grain s= ize=20 of Pd. I've never seen a sputter coater for SEM sample prep that had=20= two=20 targets mounted and ready to go in the same pump down cycle, this soun= ds=20 like a device not designed for SEM sample prep but for industrial sput= ter=20 applications. =20 =20 Pt and Cr are other common target materials that you might also consid= er,=20 however I suspect your problem is not the coating but more likely the=20 sample. You did not state what the sample was so I am guessing that t= here=20 may be some sample deformation of the sample related to vacuum=20 incompatibility or heat from the coater. This could be shrinkage from= the=20 evacuation of the chamber or heat from the magnetron sputter head. =20 If there is a possibility that the sample may be the problem, try sput= ter=20 coating your sample and a piece of metal or carbon disc at the same ti= me. =20 If the coating is the source of the artifact the artifact should be=20 observable on all of the samples. =20 =20 I have had some problems with my server, could you acknowledge receipt= of=20 this post. Thanks and good luck. =20 Hope this helps. =20 John Humenansky Braun Intertec Corp. 6875 Washington Ave. So. Minneapolis, MN 55439 (612) 942-4822 =20 =20 =20
In addition the summary of responses on the Polaron E3000 CPD from Susan Carbyn:
I agree with most of the procedures outlined in your summary and have had frequent leaks with the front seal leaks on our Polaron E3000 since 1992. The unit was put into service in 1979 and from that time until October 1992 I replaced 5 door and 8 Dowtey window seals for 275 drying runs (ave. 34 runs/window seal). Since October 1992 I have used 25 window seals for 113 drying runs (ave. 5 runs/window seal). I am still using the door seal that was installed in March 1990! It has some fine cracks but works every time.
Most recently I have had severe leak problems due to a few defective seals as the manufacturer has mentioned. I am waiting for new replacements. Last month out of desperation for another seal I installed a used one (installed in November 1990) that was removed from the window for preventative maintenance (after 38 runs) in October 1992. It still works ok after 7 more runs.
We use only ethanol, have been heating and cooling with the same system since the unit was installed and always pressurize between 22 and 19 degrees centigrade. I have looked carefully at the surface of the rubber on the newer 1990's seals compared to the used 1980's ones (I inspect all of my seals with a light microscope before installation.). The rubber on the old 80's seals is very smooth and even on the sides and edges. The seals I purchased in the 90's are striated on the sides and have irregular edges. Because of my recent experience successfully reusing my old window seal (as well as the 175 runs on my old door seal) I suspect that the rubber and seal tooling may have changed since the late 80's and could be the cause of the increased failure rate.
The only solution I have for this problem is to increase the budget for replacement seals and switch to using HMDS whenever possible.
Regards,
Jim
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility U-131 Storrs, CT 06269 bsgphy3-at-uconnvm.uconn.edu 860 486-2914 voice 860 486-1936 fax
I'm posting this for a colleague not on the list. Please respond to Daniel Wilcox at wilcox-at-mailback.macom.com
Daniel is looking for sputter targets for his Technics Hummer 5 sputter coater. (I may not have the manufacturer's name correct, but contact Daniel for exact details.) Any help finding a source is appreciated.
We would like to get super TEM of mouse cardiac muscle. The best technique I know of is to perfuse through the abdominal vena cava, aiming toward the heart, with 4%paraform/2% Glut in cacodylate buffer. Should I use saline first to flush out the blood, should I use pressure, either syringe or gravity? Should I forget the above and listen to some of your comments. I'd appreciate your advice. Thanks so much.
John Hardy City of Hope Medical Center EM Lab jhardy-at-smtplink.coh.org
Hello All, I have been asked recently if there is any way to image micelles, such as those formed by lubricant additives. I thought that light microscopy might work, and possibly confocal. Does anyone have experience in doing this? I have a feeling I will have to refer this work to an outside lab. Regards, Melanie Behrens
Thank you very much All answered the questions about optimal conditions for sputter coating. Reducing Argon pressure and voltage works great even with a gold target!
Thanks again, Alex _________________ Alexander Titkov
Millennium Inorganic Chemicals PO Box 245 Bunbury WA 6231 Australia Ph (097) 808 505 FAX: (097) 808 500 E-mail: scm!atitkov-at-scmaust.attmail.com
At 23:14 9/04/97 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
These can be imaged using cryo-TEM. Drs Katarina Edwards and Mats Almgren at Dept of Physical Chem, Uppsala University in Sweden have done lots of this type of imaging. I am a bit out of date on this but have some papers.
Gustafsson-J; Arvidson-G; Karlsson-G; Almgren-M, Complexes between cationic liposomes and DNA visualized by cryo-TEM. Biochim-Biophys-Acta. 1995 May 4; 1235(2): 305-12
Edwards et al 1989 Langmuir 5 473-378
and also Edwards and Almgren 1991 Solubilization of lecithin vesicles by C12E8- Structural transitions and temperature effects. J Colloid Interface Sci. I dont have the number for this
Mail address, Mats Almgren, Dept Physical Chemistry, Uppsala University, PO Box 532, S-751 21 Uppsala, Sweden.
this may provide a lead
Dr. Bridget Southwell Department of Anatomy and Cell Biology University of Melbourne Parkville Vic 3052 AUSTRALIA Tel: +61 3 9344-7646 Fax: +61 3 9347-5219
At 23:14 9/04/97 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
These can be imaged using cryo-TEM. Drs Katarina Edwards and Mats Almgren at Dept of Physical Chem, Uppsala University in Sweden have done lots of this type of imaging. I am a bit out of date on this but have some papers.
Gustafsson-J; Arvidson-G; Karlsson-G; Almgren-M, Complexes between cationic liposomes and DNA visualized by cryo-TEM. Biochim-Biophys-Acta. 1995 May 4; 1235(2): 305-12
Edwards et al 1989 Langmuir 5 473-378
and also Edwards and Almgren 1991 Solubilization of lecithin vesicles by C12E8- Structural transitions and temperature effects. J Colloid Interface Sci. I dont have the number for this
Mail address, Mats Almgren, Dept Physical Chemistry, Uppsala University, PO Box 532, S-751 21 Uppsala, Sweden.
this may provide a lead
Dr. Bridget Southwell Department of Anatomy and Cell Biology University of Melbourne Parkville Vic 3052 AUSTRALIA Tel: +61 3 9344-7646 Fax: +61 3 9347-5219
I am surprised to note the high frequency of door seal leaks experienced by users of the Polaron CPD apparatus. We have two such units which have been in use since the mid-1970's and we have experienced very few of these problems. The procedures we use appear to be similar to those used by the people experiencing these frequent leaks. Perhaps, therefore, Jim Romanow's suggestion that the earlier seals, or even the CPD units themselves, were less prone to failure than the newer ones is correct. Having said that we seldom experience these problems, we did have a chamber door leak a few days ago but this was solved immediately by cleaning the door seat and replacing the seal with a spare which we had. This spare was from a set of spare seals purchased in the early 1980's so was probably of the vintage which Jim suggests has a better record than the more recently-manufactured seals.
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa eurc-at-giraffe.ru.ac.za - tel: +27 461 318168 - fax: +27 461 24377
} Hello All, } I have been asked recently if there is any way to image micelles, such as } those formed by lubricant additives. I thought that light microscopy might } work, and possibly confocal. Does anyone have experience in doing this? I } have a feeling I will have to refer this work to an outside lab. } Regards, } Melanie Behrens
I've seen some nice images of this type of specimen produced using cryo-TEM. You might want to check with a lab that is in the food or cosmetics industry.
Regards,
--------------------------------------------------------------- Dr L. P. Stoter Technical Editor, MICROSCOPY & ANALYSIS Technesis 17, Rocks Park Road email: LPS-at-teknesis.demon.co.uk Uckfield, E. Sussex Phone: +44 (0)1825 766911 TN22 2AT Fax: +44 (0)1825 766911 United Kingdom ---------------------------------------------------------------
} I have been asked recently if there is any way to image micelles, such as } those formed by lubricant additives. I thought that light microscopy might } work, and possibly confocal. Does anyone have experience in doing this? I } have a feeling I will have to refer this work to an outside lab.
Bridget Southwell (B.Southwell-at-Anatomy.UniMelb.EDU.AU) suggested the use of CryoTEM and referred to the work of M. Almgren et al. Several groups have successfully applied cryoTEM to imaging micelles. You might want to look at the recent papers from Y. Talmon' group and some older work (1989-1990) by Phillip Vinson.
One caution: most of this work (and my own as well) has studied oil-in-water systems ("normal" micelles.) These systems are relatively easy to image by cryoTEM. I suspect that "lubricant additives" are water-in-oil systems. Several people have attempted cryoTEM on water-in-oil systems (inverse micelles) and have experienced problems with extreme sensitivity to radiation damage. There is some real interesting radiation chemistry that occurs when one irradiates large concentrations of hydrocarbons in the presence of vitreous ice. -- Best Regards, John Minter
Eastman Kodak Company Phone: (716) 722-3407 Analytical Technology Division FAX: (716) 477-3029 Room 2112 Bldg 49 Kodak Park Site email: minter-at-kodak.com Rochester, NY 14562-3712 calendar: via PROFS
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
John, The best fixation I've done and ever seen was to simply dice the heart in Millonigs phosphate buffered glutaraldehyde, wash in Millonigs buffer and post fix in Millonigs OsO4. Ian.
We routinely do cardiac perfusions on mice to collect various tissues. We use 2.5% glutaraldehyde in cacodylate buffer and a perfusion pump. Our procedure is as follows.
Set up the perfusion pump with 2 x 30 ml syringes (using a threeway stopcock) and a 21 gauge butterfly needle. Adjust it to deliver a constant rate of perfusate (3 ml/min for 3-6 wk old mice, 6 ml/min for mice 7 wk or older).
With the mouse well anaesthetized, hold the xyphoid process with a hemostat and cut along either side of the sternum along the rib cage allowing the chest wall to be reflected. Insert the needle into the left ventricle and cut the right atrium immediately to allow the perufsate to escape. Wash for 10 min using lactate Ringers or until 30 ml has cleared. Switch to the fixative and fix for 10 min (or 30 ml.) without retracting the needle. Remove any desired tissue samples and continue fixation for 2 h - overnight at 4 degrees C. Then process tissues as usual.
I don't know if the technique would damage the cardiac muscle that you are interested in but I would definitely flush with saline first. As to the syringe/gravity pressure question, we find that while gravity works very well for larger animals such as rats, it's hard to get the right constant pressure on small mice. If you want any more details, let me know.
Pat Hales Dept. of Anatomy & Cell Biology McGill University hales-at-hippo.medcor.mcgill.ca
} Hello All, } I have been asked recently if there is any way to image micelles, such as } those formed by lubricant additives. I thought that light microscopy might } work, and possibly confocal. Does anyone have experience in doing this? I } have a feeling I will have to refer this work to an outside lab. } Regards, } Melanie Behrens
How large? The detergents I am familiar with form micelles no larger than the size of globular proteins, so light microscopy is of little use.
If they are large enough for light microscopy, I would try differential interference contrast (Nomarski), which should yield a good image of the edge of the micelle against the background.
Jim Williams.
/////////////////////////////////////////////////////////////////////////// / James C. Williams, Jr. williams-at-anatomy.iupui.edu / / Department of Anatomy / / Indiana University School of Medicine (317)274-3423 / / 635 Barnhill Drive (317)278-2040 fax / / Indianapolis, IN 46202-5120 / /////////////////////////////////////////////////////////////////////////// Great are the works of the LORD, studied by all who have pleasure in them. Psalm 111:2
} We would like to get super TEM of mouse cardiac muscle. The best } technique I know of is to perfuse through the abdominal vena cava, } aiming toward the heart, with 4%paraform/2% Glut in cacodylate buffer. } Should I use saline first to flush out the blood, should I use } pressure, either syringe or gravity? Should I forget the above and } listen to some of your comments. I'd appreciate your advice. Thanks } so much. } } John Hardy } City of Hope Medical Center } EM Lab } jhardy-at-smtplink.coh.org
John- Definitely flush first, otherwise the blood will coagulate in the fix and block the flow of blood to the heart. You don't say what animal you are using, but in general any vessel close to and with as direct as possible access to the vessels of interest can be used for perfusion. I have had good luck in perfusing the coronary arteries and thus also the heart muscle in rats by inserting the perfusion needle directly into the left ventricle. This has the added advantage of distributing fixative to the inner wall of the heart. We inject with heparin just before anesthesia as an added measure against coagulation during manipulation, cannulation, and before flushing of the artery is complete. I prefer to flush with the same buffer used for the fixative (cacodylate) rather than saline. I don't have imperical data, however, to prove one type of flush is better than another. We often do enzyme or immunohistochemistry on tissue and my protocols are often based on the "if it ain't broke don't fix it" model and my belief that the best histochemist are really voodoo priests in disguise.
I hope this helps Jay ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-bgsm.edu * **************************************************************
Thank you to everyone who has sent information about Hummer targets. Daniel Wilcox said that his network has been experiencing problems, so I have been forwarding messages sent to me. My e-mail address is audrey.dow-at-amp.com
In response to the following: ============================================== I have been asked recently if there is any way to image micelles, such as those formed by lubricant additives. I thought that light microscopy might work, and possibly confocal. Does anyone have experience in doing this? I have a feeling I will have to refer this work to an outside lab. ============================================== One of the earliest pieces of work I can remember being published came out of the old Sinclair Research Laboratories on the South Side of Chicago. I regret I can not remember names, because some of their work was published (some not) in the early 1960's but they had beautiful images of features from lubricants, having the appearance sometimes of almost filamentous like structures. These were before the days of SEM and hence, the work was all done by Pt/C replication techniques and TEM. Note: Even if SEM was around, the dimensions of the structures were such that the features would have been difficult to impossible to resolve anyhow. Off line, if anyone is interested, I would related more on the history of how the technique was developed, which was also influenced by Mr. Bill Ladd, founder of Ladd Reserach Industries, Inc.
So if the Pt/C replication approach will show what you are looking for, it will be faster and cheaper than any other method. Now I am not sure that these filamentous structures would qualify strictly speaking as "micelles", but more often than not, in our own lab, when our clients ask to see the "micellar structure", in these kinds of materials, this is what they often times mean, because they pull out some old micrographs showing the filamentous structures! In any case, we have been practicing this technique ever since the early 1970's on lublicant samples of various types.
If what you want to see is more along the lines of a traditional micellar structure, then you have to turn to freeze fracture TEM, however, the nature of the organics present, and the need to clean off the organics (often times polymers found in today's lubricant systems) that "stick" to the replica make this approach often times a lot more tricky than might originally meet the eye. It can be, in others words, an art unto itself. But structures can be seen, they tend to be (at least the ones we have seen) an almost "network" or "fishnet" kind of structure. Maybe other structures are present as well, but these are the kinds of structures we have seen ourselves.
Disclaimer: Our firm, Structure Probe, Inc. performs these kinds of analyses as an analytical laboratory service.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
PLEASE DO NOT REPLY TO THIS E-MAIL NOR TO THE LISTSERVER
I apologise in advance to those this inconveniences, but I know that many potential MICROSCOPY & ANALYSIS readers subscribe to these lists, and want to make sure they get this information.
1. Coverage of the USA edition of M&A will soon be extended to include Canada.
2. A new Asia/Pacifc edition of M&A will be launched this Autumn. This will cover approximately the region India to the Philippines and Japan to Australia.
MICROSCOPY & ANALYSIS is mailed FREE to qualified readers, at business addresses. Qualified means you purchase or specify microscopy and related instrumentation.
If you are in Canada or the Asia/Pacific region (or the USA or Europe and don't yet get MICROSCOPY & ANALYSIS), please e-mail Lynda Stowell {Lynda-at-microrgc.demon.co.uk} for further details about obtaining a subscription.
Alternatively, our new WWW site will be activated in the next 2-3 weeks, and will include a subscription form. The address for the site will be http://www.microrgc.demon.co.uk. THE SITE IS NOT YET ACTIVE.
Regards,
--------------------------------------------------------------- Dr L. P. Stoter Technical Editor, MICROSCOPY & ANALYSIS Technesis 17, Rocks Park Road email: LPS-at-teknesis.demon.co.uk Uckfield, E. Sussex Phone: +44 (0)1825 766911 TN22 2AT Fax: +44 (0)1825 766911 United Kingdom ---------------------------------------------------------------
} Hello All, } I have been asked recently if there is any way to image micelles, such as } those formed by lubricant additives. I thought that light microscopy might } work, and possibly confocal. Does anyone have experience in doing this? I } have a feeling I will have to refer this work to an outside lab. } Regards, } Melanie Behrens
Cryotechnique is the way to go, but "cryo-TEM" isn't the best approach. I recommend freeze-fracture; see Robards & Sletyr, "Low temperature methods in biological EM" (v. 10 of the Glauert series), 1985, and Zazadzinski & Bailey,"Freeze-fracture of polymers", J.E.M. Technique 13:309-334(1989). There are commercial labs with the necessary equipment.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 http://www.MSA.microscopy.com/ProjectMICRO/Books.html
I am new to this list and am trying to do some immunogold labeling studies (I am also a novice in this area). I have a problem that I can't figure out and thought someone out there could help me. It is probably a simple one, but so far I can not find a solution. I am working with samples that are pretty fragile and found that I need to place them on coated grids for stability. I am coating my nickel grids with Pioloform and have found that my sample looks good on these grids. I tried to immunogold label and found non specific labeling throughout the sample, on the resin, and on the grid itself. I have now done tests on the coated grid alone and have processesd it as I would normally. I find that I have gold label everywhere on the coated grid. Therefore I have concluded that an interaction between my primary antibody and the coating is occuring (I have done a control experiment leaving out the primary antibody in the processing and know nothing binds). What could cause this? My blocking solution consists of 1% BSA in TBS +.05% Tween 20 + .02% azide for preservation. Could it be that the blocking solution is not doing its job? How long can you store blocking solution? Could I be using a too concentrated antibody solution? Could this be specific to my antibody? I should mention that I wash my grids by either passing them through drops of TBS or wash using a light stream of TBS from a pasteur pipet. Both methods yield the same results. If anyone can give me some answers to my questions I would appreciate it. Thanks for your time.
I'm looking for suppliers of Ellis type fiberoptic light scramblers. I have found one company, Technical Video in Woods Hole. Are there others you can recommend?
We routinely do cardiac perfusions on mice to collect various tissues. We use 2.5% glutaraldehyde in cacodylate buffer and a perfusion pump. Our procedure is as follows.
Set up the perfusion pump with 2 x 30 ml syringes (using a threeway stopcock) and a 21 gauge butterfly needle. Adjust it to deliver a constant rate of perfusate (3 ml/min for 3-6 wk old mice, 6 ml/min for mice 7 wk or older).
With the mouse well anaesthetized, hold the xyphoid process with a hemostat and cut along either side of the sternum along the rib cage allowing the chest wall to be reflected. Insert the needle into the left ventricle and cut the right atrium immediately to allow the perufsate to escape. Wash for 10 min using lactate Ringers or until 30 ml has cleared. Switch to the fixative and fix for 10 min (or 30 ml.) without retracting the needle. Remove any desired tissue samples and continue fixation for 2 h - overnight at 4 degrees C. Then process tissues as usual.
I don't know if the technique would damage the cardiac muscle that you are interested in but I would definitely flush with saline first. As to the syringe/gravity pressure question, we find that while gravity works very well for larger animals such as rats, it's hard to get the right constant pressure on small mice. If you want any more details, let me know.
Pat Hales Dept. of Anatomy & Cell Biology McGill University hales-at-hippo.medcor.mcgill.ca
Caroline Schooley recommended that Melanie Behrens use freeze fracture to image micelles in her lubricant's. Our lab did some comparisons between freeze-fracture TEM and direct imaging of vitrified micelles of cetyltrimethylammonium chloride (CTACl)/ sodium salicylate (NaSal). Both the CTACl globular micelles and worm-like micelles formed with the addition of NaSal to CTACl are easily imaged by direct cryoTEM. We were unable to obtain contrast in freeze-fracture images without etching. The structures observed in freeze-fracture/freeze-etch micrographs of these systems were much larger than those observed by direct cryoTEM. ALWAYS beware of changes upon etching these microstructures. Our conclusions were that the freeze-fracture, freeze-etch micrographs were dominated by artifacts. I'd be interested in hearing from anyone who had obtained convincing evidence of micellar microstructures by freeze-frature.
-- Best Regards, John Minter
Eastman Kodak Company Phone: (716) 722-3407 Analytical Technology Division FAX: (716) 477-3029 Room 2112 Bldg 49 Kodak Park Site email: minter-at-kodak.com Rochester, NY 14562-3712 calendar: via PROFS
Hello microscopists, I am in the process of assembling an ultra high vacuum chamber with mag. lev. turbo pumps backed by oil free diaphragm pumps. Some parts e.g. gauges on this system will come from other decommissioned systems. I am wondering if someone would like to share experience in: 1. converting viton onto metal O-ring sealings as compared to redesigning flanges to accomodate KFs; including costs, companies, and suppliers; 2. reliability, service record, and major troubles with Leybold, Balzers/Pfeiffer, Osaka, etc magnetic bearing turbo pumps. Sincerely, Marek Malecki.
This should be a simple procedure but I am having a terrible time trying to stain some Embed 812 embedded tissues with uranyl acetate/lead citrate. The tissue is from frog tadpoles, fixed in glutaraldehye/paraformaldehyde and postfixed in osmium. I have tried staining for 5-20 minutes with saturated solution of UA in ethanol, followed by 1-5 minutes in lead citrate (Venable-Coggeshall formulation). The sections look no different from unstained specimens.
I'd appreciate any suggestions about what I might be doing wrong.
Gary Radice, Associate Professor gradice-at-richmond.edu Department of Biology 804-289-8107 (voice) University of Richmond VA 23173 804-289-8233 (FAX)
Leitz has built one several years ago (called Variolum) for their Diaplan, Aristoplan microscopes. I have one of these. Although intended for the regulation of light intensity of xenon and Hg arcs it performs well in video microscopy. You should contact Leitz, or some part dealers to find one.
Does anyone have a good protocol for embedding tadpole brain for doing immunocytochemistry? We have been trying and seem to be having a problem with infiltration. Please let me know. Thanks in advance.
Mei Lie Wong Department of Biochemistry HHMI-UCSF Ph. 415-476-4441 Fax 415-476-1902 email wong-at-msg.ucsf.edu
Dear Susan, You do not say in your message what the antibodies are directed against. One very obvious possibility is that there is specific reactivity to BSA. This would, of course, result in very specific binding to the BSA covering the sample, the resin and the film. This would look as if there was non-specific binding.
A simple test is to change your blocking agent. Try 1% cold-water fish skin gelatin (very cheap, from Sigma) in PBS as a substitute. It is useful because there are no mamalian serum proteins present and for anyone interested in localizing biotin with antibodies, there are no biotin-like molecules present.
If changing the blocking agent doesn't work, I would suspect that the antibody specifically reacts with the resin and the plastic (just kidding!).
There should be no non-specific reactions between the antibodies and the plastic film or resin. Suspect instead the blocking agent or as a last resort, the antibody.
I have included a section on reasons why antibody labeling may not work on my web site.
regards,
Paul Webster, Ph.D. Center for Cell Imaging Yale School of Medicine http://info.med.yale.edu/cellimg --------------------------------------
Hello,
I am new to this list and am trying to do some immunogold labeling studies (I am also a novice in this area). I have a problem that I can't figure out and thought someone out there could help me. It is probably a simple one, but so far I can not find a solution. I am working with samples that are pretty fragile and found that I need to place them on coated grids for stability. I am coating my nickel grids with Pioloform and have found that my sample looks good on these grids. I tried to immunogold label and found non specific labeling throughout the sample, on the resin, and on the grid itself. I have now done tests on the coated grid alone and have processesd it as I would normally. I find that I have gold label everywhere on the coated grid. Therefore I have concluded that an interaction between my primary antibody and the coating is occuring (I have done a control experiment leaving out the primary antibody in the processing and know nothing binds). What could cause this? My blocking solution consists of 1% BSA in TBS +.05% Tween 20 + .02% azide for preservation. Could it be that the blocking solution is not doing its job? How long can you store blocking solution? Could I be using a too concentrated antibody solution? Could this be specific to my antibody? I should mention that I wash my grids by either passing them through drops of TBS or wash using a light stream of TBS from a pasteur pipet. Both methods yield the same results. If anyone can give me some answers to my questions I would appreciate it. Thanks for your time.
Susan
------------------ RFC822 Header Follows ------------------ Received: by QuickMail.Yale.edu with SMTP;10 Apr 1997 17:27:25 -0400 Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by mail-relay1.cis.yale.edu (8.8.5/8.8.5) with SMTP id RAA08528 for {Paul.Webster-at-Yale.edu} ; Thu, 10 Apr 1997 17:25:29 -0400 (EDT) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id MAA06215 for dist-Microscopy; Thu, 10 Apr 1997 12:11:39 -0500 Received: from pilot12.cl.msu.edu (pilot12.cl.msu.edu [35.9.5.22]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id MAA06212 for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 10 Apr 1997 12:11:37 -0500 Received: from 35.8.197.89 ([35.8.197.89]) by pilot12.cl.msu.edu (8.7.5/MSU-2.10) id NAA11591; Thu, 10 Apr 1997 13:11:16 -0400 Message-ID: {334D3BB4.7153-at-pilot.msu.edu}
Hello!
We have had several things to cause this in our sections:
1. Specimen was not osmicated enough, OsO4 too dilute, or bad.
2. or... The lead stain was not the right ph and actually bleached the sample { goal~ = 12 pH} ( we made a switch in Lead stain types just 2 months ago for this very reason.)
3. or.... Some epoxy mixtures, if not polymerized enough or infiltrated ( acts like a really soft block) then the thicks will overstain and the thins will understain. In doing some epoxy mixture experiments, I found with the same chemicals, that certain ratios caused some more hard to stain sections.
4. or..... someone made a mistake by a factor of 10 in the NaOH [ ] of 1st rinse dip after lead stain, and it bleached the dickens out of the section. We had a researcher do this to their sections.
5. or..... we have bad water or something in our building, our aquous Uranyl Acetate only does well for 10 days, so we make it up in very very small amounts. I like the clearer background we get with aqueous much better than the alcohol methods, but that is personal preference.
6. I've developed a microwave technique, and my grids stain really well now. Write if you want to hear more....it's too lengthy for this message.
We stopped using Epon812 about 7 years ago, because at the time it had a lot of impurities in it, the resin ate up our diamond knives. And the blocks were real difficult, poor polymerization despite how we would polymerize it. Switched to Lx112 from Ladd and our diamonds last 6 X longer.
Hope some of this helps, Lou Ann
} This should be a simple procedure but I am having a terrible time trying to } stain some Embed 812 embedded tissues with uranyl acetate/lead citrate. The } tissue is from frog tadpoles, fixed in glutaraldehye/paraformaldehyde and } postfixed in osmium. I have tried staining for 5-20 minutes with saturated } solution of UA in ethanol, followed by 1-5 minutes in lead citrate } (Venable-Coggeshall formulation). The sections look no different from } unstained specimens. } } I'd appreciate any suggestions about what I might be doing wrong. } } } Gary Radice, Associate Professor gradice-at-richmond.edu } Department of Biology 804-289-8107 (voice) } University of Richmond VA 23173 804-289-8233 (FAX)
Lou Ann Miller Center for Microscopy & Imaging College of Veterinary Medicine Dept. of Veterinary Biosciences University of Illinois Rm 1108 Basic Sciences Bld 2001 S Lincoln Ave. Urbana, Illinois 61802
Phone: 217-244-1567 email: lamiller-at-uiuc.edu
Center for Microscopy & Imaging Home page: http://www.cvm.uiuc.edu/MicImagLab/MicImagLab.html
Central States Microscopy Society: http://www.cvm.uiuc.edu/Homepages/LouAnnMiller/CSMS/csms.html
Personal Home Pages: http://www.cvm.uiuc.edu/Homepages/LouAnnMiller/LAM.html
Attention ESEM owners and users: If you were desperate for the low magnification in ESEM (E3, 2020) in a HiVac mode do not worry from now on. Simply, take out the rotation/tilt module and put your specimen directly on the driving assembly. You will be deprived of rotation and tilting of a specimen but you will gain extra 62 mm of working distance. Alternatively, put you sample in a brass or aluminium cylinder with a side screw to keep the specimen firm and glue the cylinder on the driving assembly with a double-sided conductive tape. The height of cylinder is such that it can accomodate a full lenght of a pin of a pin-type stub. In this case you will gain 'only' about 50 mm of working distance. Even without playing with apertures you should get minimum mag ~6X at acc. voltage of 2 kV, covering an area of 13 mm in diameter with a much better collecting angle for SE, when using ET detector. Try for yourself and enjoy the results. Cheers, Wis Jablonski, OiC EM/X-Ray Microanalysis, Uni of Tasmania
Hello! A student at our department shall try to label membrane proteins on mouse spermatozoa for study with SEM. We have access to cryo SEM as well which might be the fastest procedure. Conventional technique with chemical fixation and labelling of single cells or label fresh (living) spermatozoa and then use cryotechniques? Any hints will be appreciated.
Hans Ekwall Centre for Reproductive Biology Dept. of Anatomy & Histology SLU, Box 7011, 75007 Uppsala SWEDEN
Has anyone comments on the UV polimeriztion of Lowicryl HM20 after freeze substitution in Acetone-Uranylacetate. I have problems with prepolymerization of the resin which causes unusable blocks, but the problem is not consistent, some blocks are good some are not.
TIA, Stefan
Dr. Stefan Hillmer Albrecht-von-Haller Institut fuer Pflanzenwissenschaften Universitaet Goettingen Untere Karspuele 2 37073 Goettingen Deutschland
Tel (+49) 551-392013 Fax (+49) 551-397833 e-mail shillme-at-gwdg.de
we have had similar problems on pioloform coated grids, and I would say that your 1. AB is binding to the pioloform. Solution: Try change the coating to formvar or a carbon coating. You can also make the grids float on the 1. AB so that the ABs only get in contact with the pioloform outside the sections.
Bo
} Hello, } } I am new to this list and am trying to do some immunogold labeling } studies (I am also a novice in this area). I have a problem that I } can't figure out and thought someone out there could help me. It is } probably a simple one, but so far I can not find a solution. I am } working with samples that are pretty fragile and found that I need to } place them on coated grids for stability. I am coating my nickel grids } with Pioloform and have found that my sample looks good on these grids. } I tried to immunogold label and found non specific labeling throughout } the sample, on the resin, and on the grid itself. I have now done tests } on the coated grid alone and have processesd it as I would normally. I } find that I have gold label everywhere on the coated grid. Therefore I } have concluded that an interaction between my primary antibody and the } coating is occuring (I have done a control experiment leaving out the } primary antibody in the processing and know nothing binds). What could } cause this? My blocking solution consists of 1% BSA in TBS +.05% Tween } 20 + .02% azide for preservation. Could it be that the blocking solution } is not doing its job? How long can you store blocking solution? Could I } be using a too concentrated antibody solution? Could this be specific to } my antibody? I should mention that I wash my grids by either passing } them through drops of TBS or wash using a light stream of TBS from a } pasteur pipet. Both methods yield the same results. If anyone can give } me some answers to my questions I would appreciate it. Thanks for your } time. } } Susan } }
Advanced International Immunofluorescence Course Gargnano '97 (Italy)
The Advanced International Immunofluorescence Course is a post-doctorate theoretical/practical course, with propedeutical lectures and practical stages on traditional and confocal immunofluorescence microscopy and image and ion analysis. The course will take place in Gargnano (Lake of Garda) from 7 to 10 October 1997. Further information and registration details will be found at the following Web address
http://imiucca.csi.unimi.it/endomi/ACIF.html
Thank you Paolo Castano ____________________________________________________________________________ _______
Prof. Paolo Castano UNIVERSITY OF MILAN INSTITUTE OF HUMAN ANATOMY - CHAIR OF HUMAN ANATOMY FOR PHARMACY Via Mangiagalli, 31 - 20133 Milan (Italy)
} I have been asked recently if there is any way to image micelles, such as } those formed by lubricant additives. I thought that light microscopy might } work, and possibly confocal. Does anyone have experience in doing this? I } have a feeling I will have to refer this work to an outside lab.
Bridget Southwell (B.Southwell-at-Anatomy.UniMelb.EDU.AU) suggested the use of CryoTEM and referred to the work of M. Almgren et al. Several groups have successfully applied cryoTEM to imaging micelles. You might want to look at the recent papers from Y. Talmon' group and some older work (1989-1990) by Phillip Vinson.
One caution: most of this work (and my own as well) has studied oil-in-water systems ("normal" micelles.) These systems are relatively easy to image by cryoTEM. I suspect that "lubricant additives" are water-in-oil systems. Several people have attempted cryoTEM on water-in-oil systems (inverse micelles) and have experienced problems with extreme sensitivity to radiation damage. There is some real interesting radiation chemistry that occurs when one irradiates large concentrations of hydrocarbons in the presence of vitreous ice. -- Best Regards, John Minter
Eastman Kodak Company Phone: (716) 722-3407 Analytical Technology Division FAX: (716) 477-3029 Room 2112 Bldg 49 Kodak Park Site email: minter-at-kodak.com Rochester, NY 14562-3712 calendar: via PROFS
Does anyone have any thoughts or experience with using a modern (e.g. Generation 3) image intensifier tube to enhance CCD imaging on a TEM? I don't know the limitations of image intensifiers (e.g. noise, resolution) but the potential for increased gain on high-resolution, short exposure time shots might be worth investigating.
Daniel L. Callahan Mechanical Engineering and Materials Science Rice University, MS 321 6100 S. Main St Houston, TX 77005-1892
Thanks to all who have responded to the call for lead speciation in smelter vs natural occuring products. The information has been forwarded to the correct person and they will make there decision soon because the final report is due in 5 weeks. Thanks again.
Clarissa Vierrether The Doe Run Co. cvierret-at-misn.com
I have never tried Pioloform but routinly use either parlodion coated nickel or formvar coated nickel grids without backround problems. That would be the first thing I would test. The other possiblity is that the original antigen was bound to BSA as a stablizer when they made the antibody and now the primary is binding to the BSA. Try using purified gelatin instead of BSA for blocking.
On Thu, 10 Apr 1997, Susan Fujimoto wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello, } } I am new to this list and am trying to do some immunogold labeling } studies (I am also a novice in this area). I have a problem that I } can't figure out and thought someone out there could help me. It is } probably a simple one, but so far I can not find a solution. I am } working with samples that are pretty fragile and found that I need to } place them on coated grids for stability. I am coating my nickel grids } with Pioloform and have found that my sample looks good on these grids. } I tried to immunogold label and found non specific labeling throughout } the sample, on the resin, and on the grid itself. I have now done tests } on the coated grid alone and have processesd it as I would normally. I } find that I have gold label everywhere on the coated grid. Therefore I } have concluded that an interaction between my primary antibody and the } coating is occuring (I have done a control experiment leaving out the } primary antibody in the processing and know nothing binds). What could } cause this? My blocking solution consists of 1% BSA in TBS +.05% Tween } 20 + .02% azide for preservation. Could it be that the blocking solution } is not doing its job? How long can you store blocking solution? Could I } be using a too concentrated antibody solution? Could this be specific to } my antibody? I should mention that I wash my grids by either passing } them through drops of TBS or wash using a light stream of TBS from a } pasteur pipet. Both methods yield the same results. If anyone can give } me some answers to my questions I would appreciate it. Thanks for your } time. } } Susan }
SHORT COURSE ANNOUNCEMENT ------------------------------------------------- FUNDAMENTALS AND APPLICATIONS OF LIGHT MICROSCOPY ------------------------------------------------- JUNE 22-27, 1997, Burlington Vermont
Experienced microscopy problem solvers will teach a 5-day course on achieving the maximum information from light microscopy. The emphasis of the course will be to provide hands-on experience, and the background for interpretation of images.
The course will cover the principals of light microscopy, contrast techniques for the microscope, adjustments of the microscope for optimum contrast and resolution, interpretation of images in terms of light-matter interactions, and image recording.
A full range of reflected and transmitted light microscopes, as well as contrast accessories, will be provided for use by the students. Students are encouraged to bring their own samples.
Faculty: Philip C. Robinson, author of the RMS Microscopy Series book, "Applied Polarized Light Microscopy", Robert Janes, of the Metropolitan Forensic Science Laboratory, and Mary. McCann, McCann Imaging, course organizer. Vermont Optecs, specialists in research instruments, will supply microscope equipment for the course.
For course brochure and registration information, contact Mary McCann, McCann Imaging e-mail: mccanns-at-tiac.net telephone 617-484-7865 fax: 617-484-7865 Further information: www.microscopyed.com
The french company ALTO MEDIA is producing a series of short movies (3 '), based on a 'travel' into the matter... 5 materials have already been investigated : stell, aluminium, alumina, concrete and brass. These movies require that sequences of both SEM and TEM images are realized. They will be distributed in France (Cite des Sciences, TV broadcasts), and could also be distributed in other european countries (Italy, U.K.,...). ALTO MEDIA is looking for partners, i.e. microscopists, who could be interested to collaborate to this project. The following (non-exhaustive) list gives some ideas of possible interesting materials :
paper/wood/porcelain/plaster/skin/bone/ stone/diamond/graphite/plastics/silicom/iron(rust?)ice/composites(kevlar,mylar)/ ice/glues/lubricating oils,..., and all kinds of materials that are currently used, and for which a microscopic observation can help to understand their properties.
Contact : ALTO MEDIA Etienne Blanchon or Gabriel Turquier tel: 33 01 42 77 77 72 Fax : 33 01 42 77 77 73 e-mail : altomail-at-worlnet. fr
--------------------------------------------------------------------------- Dr. Thierry EPICIER, GEMPPM, umr CNRS 5510, INSA de LYON, Bat 502, 20, Av. Einstein, F69621 VILLEURBANNE CEDEX FRANCE
Hi Gary, We use Embed 812 to process retina tissue. We get intense staining using 4% Uranyl Acetate in 100% methanol.Stain for 30 or 40 minutes at room temp. Keep solution in dark.Rinse in 3 changes of methanol. Then counter stain with Reynold's Lead Citrate for 1-10 minutes. Rinse with 3 changes of DI water. We get beautiful stain if the tissue is well fixed.
Sally
On Thu, 10 Apr 1997, Gary Radice wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } This should be a simple procedure but I am having a terrible time trying to } stain some Embed 812 embedded tissues with uranyl acetate/lead citrate. The } tissue is from frog tadpoles, fixed in glutaraldehye/paraformaldehyde and } postfixed in osmium. I have tried staining for 5-20 minutes with saturated } solution of UA in ethanol, followed by 1-5 minutes in lead citrate } (Venable-Coggeshall formulation). The sections look no different from } unstained specimens. } } I'd appreciate any suggestions about what I might be doing wrong. } } } Gary Radice, Associate Professor gradice-at-richmond.edu } Department of Biology 804-289-8107 (voice) } University of Richmond VA 23173 804-289-8233 (FAX) } } }
Check the following things regarding your problem with prepolymerization of Lowicryl HM20:
1. Are all components being well mixed? The best way to mix the components is to gently bubble nitrogen gas through a glass pipette tip for about 15-20 minutes, or until the benzoyl peroxide catalyst is completely dissolved.
2. Is UV light reaching all sides of the capsules? If you are using polyethylene capsules, are they suspended in wire loops so they are not irradiated from just the tops or the sides?
3. Is the proper UV light being used? Use long-wavelength (366 nm) UV. Short wavelength UV can be too energetic.
4. I would suspect the UV light is too intense. Is the UV light being properly diffused, so it is indirect? Along with this, is the UV source far enough away from the capsules? This depends on the intensity of the UV source, but the UV light should probably be 25-30 cm above the capsules. Also, is there a reflector between the UV source and the capsules, and are all of the inner surfaces of your polymerization chamber lined with aluminum foil? Try a set-up like the following (in cross-section):
O ---------UV source /\ / \ ---------Reflector
uuu ---------Capsules
The easiest way to decrease the intensity of the UV light is to increase the distance between the bulb and the capsules. If you can't do this because of the dimensions inside the freezer or cold box, you can put a layer or two of thin, "frosted" glass in front of the UV light.
I hope this helps. Let us know how things work out.
Best regards, Bob Chiovetti (RCHIOVETTI-at-aol.com)
The staining with heavy metals (Pb, UA) is dependent on proper processing protocols to some extent: Osmium acts as a mordant for UA, and UA acts as a mordant for Pb. Sections which have been exposed to the electron beam are so highly crosslinked due to the high temperature achieved that they may never stain. (Blocks which are overpolymerized in a microwave oven will NEVER stain, no matter what one does) The most likely problem is your Pb stain and your rinsing afterwards. If the pH of the lead stain is over 12 (pH meters are not accurate at these high readings, so one cannot depend on them) there will be NO staining. NEVER, NEVER, NEVER, rinse sections which have just left the lead stain in ANY type of sodium hydroxide solution. This may (will) change the pH radically and erase any stain you may have, or it may cause your stain to "dump". Use plain water. MAKE STAIN CORRECTLY. Use the Reynolds method for lead citrate. NEVER, NEVER, NEVER, use sodium hydroxide pellets to produce the stain. Use commercially titrated sodium hydroxide ( 1.N). Pellets will NOT give you an accurate pH. (and your pH meter is no help, because it functions poorly with this unbuffered solution). Shake and invert the solution for 25 minutes. Boring? Yes! Do it anyway. It is necessary for correct chealation. A lot of information like this is to be found in my chapter in a textbook that came out last year. If you continue to have trouble, please E-mail me, and I will send you a copy of the chapter. ( hcrowley-at-DU.edu ) This type of problem makes one rabid. It drove me crazy. I finally did 4 years of work and observations until I got to the bottom of it. I found that 99% of the time it was the Pb that was so difficult to understand. Good luck. If things do not straighten up, let me know. I would be happy to help you - I know how screaming frustrating it is to have sections which do not stain. Bye, H.
The staining with heavy metals (Pb, UA) is dependent on proper processing protocols to some extent: Osmium acts as a mordant for UA, and UA acts as a mordant for Pb. Sections which have been exposed to the electron beam are so highly crosslinked due to the high temperature achieved that they may never stain. (Blocks which are overpolymerized in a microwave oven will NEVER stain, no matter what one does) The most likely problem is your Pb stain and your rinsing afterwards. If the pH of the lead stain is over 12 (pH meters are not accurate at these high readings, so one cannot depend on them) there will be NO staining. NEVER, NEVER, NEVER, rinse sections which have just left the lead stain in ANY type of sodium hydroxide solution. This may (will) change the pH radically and erase any stain you may have, or it may cause your stain to "dump". Use plain water. MAKE STAIN CORRECTLY. Use the Reynolds method for lead citrate. NEVER, NEVER, NEVER, use sodium hydroxide pellets to produce the stain. Use commercially titrated sodium hydroxide ( 1.N). Pellets will NOT give you an accurate pH. (and your pH meter is no help, because it functions poorly with this unbuffered solution). Shake and invert the solution for 25 minutes. Boring? Yes! Do it anyway. It is necessary for correct chealation. A lot of information like this is to be found in my chapter in a textbook that came out last year. If you continue to have trouble, please E-mail me, and I will send you a copy of the chapter. ( hcrowley-at-DU.edu ) This type of problem makes one rabid. It drove me crazy. I finally did 4 years of work and observations until I got to the bottom of it. I found that 99% of the time it was the Pb that was so difficult to understand. Good luck. If things do not straighten up, let me know. I would be happy to help you - I know how screaming frustrating it is to have sections which do not stain. Bye, H.
Thanks to everyone who offered suggestions for improving my UA/Pb staining of Embed 812 embedded tissues. I now have more staining protocols than I have specimens!
Among the suggestions were
en bloc stain with UA before embedding make up UA in methanol make sure the UA is really saturated by sonicating it make sure tissue is well osmicated check Pb pH is } 12 Try different lead formulation try different epoxy component proportions change to different embedding medium Increase stain time to 30 min (UA) and 5 min(Pb) Try Pb then UA then Pb Try permanganate instead of UA reduce wash times try a microwave stain protocol
Thanks to:
Lou Ann Miller Phil Oshel Robin Cross Julian Smith Tamara Howard David Patton
Gary Radice, Associate Professor gradice-at-richmond.edu Department of Biology 804-289-8107 (voice) University of Richmond VA 23173 804-289-8233 (FAX)
Wis and other ESEM users It is true that longer working distances and lower accelerating voltages in the ESEM give lower mag images and as you point out this is mainly usable at hivac. This is because in wet mode the scattering effect of the gas on the beam is much worse for both longer working distance and lower accelerating voltage dramatically lowering the image contrast to the point that it may not be useable. If the sample is compatible with hivac and you have the luxury of access to other SEMs why not use a conventional SEM? Also at hivac the final pressure limiting aperture in the ESEM can be enlarged such that it does not restrict the scanned beam at low magnification. Further Gene Taylor has come up with a low magnification device which is described in our paper: M.E. Taylor and S.A. Wight "A New Method for Low-Magnification in the Environmental Scanning Electron Microscope" SCANNING Vol. 18, 483-489 (1996). This paper discusses and compares several approaches for attaining low mag. Please see figures 3 and 4 for an explanation and demonstration of the effect of working distance on wet mode imaging.
Scott Wight
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------------------------------ Scott Wight e-mail: SCOTT.WIGHT-at-NIST.GOV NIST - Microanalysis Group W voice: 301-975-3949 Bld 222, Rm A113 | fax:301-216-1134/301-417-1321 Gaithersburg, MD 20899 \|/ disclaimer: Any opinion expressed is my own and does not represent those of my employer.
My maglev turbo experience is limited to the Leybold 340 l/s pump. I do not
recall when, exactly, I put this pump on my system, but it was shortly after
they became available (} 5 years?). This pump has been running 24 hours/day since, experiencing about 10-15 short time shutdowns from causes like power outages. Shutdown/start-up (according to Leybold - I believe 'em) is harder on the pump than continuous operation. Although it was expensive, it has performed well with zero down-time from failure. I did like the Leybold feature of not needing batteries for "spin-down". During shutdown, it uses the rotor/motor for a generator to supply magnetic bearing power and, thus, apply dynamic braking. BTW....I cannot detect any induced vibration on my rather vibration sensitive Etec SEM.
No vested interest in any pump manufacturer....
Woody White ______________________________ Reply Separator _________________________________
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello microscopists, I am in the process of assembling an ultra high vacuum chamber with mag. lev. turbo pumps backed by oil free diaphragm pumps. Some parts e.g. gauges on this system will come from other decommissioned systems. I am wondering if someone would like to share experience in: 1. converting viton onto metal O-ring sealings as compared to redesigning flanges to accomodate KFs; including costs, companies, and suppliers; 2. reliability, service record, and major troubles with Leybold, Balzers/Pfeiffer, Osaka, etc magnetic bearing turbo pumps. Sincerely, Marek Malecki.
2:00 5:00 Image analysis workshop, demonstration. Dr. Charlie Meshul's lab. V.A. Bldg. 100, Room 2C-150.
5:15 6:30 Social at Center for Research on Occupational and Environmental Toxicology / Basic Science Building's Atrium.
A quick note for those of you not familiar with our society's meeting format. There is no registration fee. Your annual dues cover registration and also the Friday night social. The social will feature cheeses, cold cuts, breads and fresh fruit and to wash it all down we have an outstanding selection of Northwest wines chosen personally by Charlie Meshul,. Make it a point to attend.
If you have any questions give Bob Mixon, Charlie Meshul, or me a call. There is parking at the VA as noted on the map.
There are tables available for any vendors that attend. If any vendor wants me to put out their literature or samples I am more than happy to help, just let one of the officers know.
Thanks to everyone who offered suggestions for improving my UA/Pb staining of Embed 812 embedded tissues. I now have more staining protocols than I have specimens!
Among the suggestions were
en bloc stain with UA before embedding make up UA in methanol make sure the UA is really saturated by sonicating it make sure tissue is well osmicated check Pb pH is } 12 Try different lead formulation try different epoxy component proportions change to different embedding medium Increase stain time to 30 min (UA) and 5 min(Pb) Try Pb then UA then Pb Try permanganate instead of UA reduce wash times try a microwave stain protocol
Thanks to:
Lou Ann Miller Phil Oshel Robin Cross Julian Smith Tamara Howard David Patton Sally Shrom Krystyna Rybicka
Gary Radice, Associate Professor gradice-at-richmond.edu Department of Biology 804-289-8107 (voice) University of Richmond VA 23173 804-289-8233 (FAX)
Does anyone have any thoughts or experience with using a modern (e.g. Generation 3) image intensifier tube to enhance CCD imaging on a TEM? I don't know the limitations of image intensifiers (e.g. noise, resolution) but the potential for increased gain on high-resolution, short exposure time shots might be worth investigating. Daniel L. Callahan Mechanical Engineering and Materials Science Rice University, MS 321 6100 S. Main St Houston, TX 77005-1892
In addition to all of the other advice you have received, thiner sections are more difficult to stain than thicker sections are; less material to attach lead and uranium to, thus less contrast.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (908)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
Available: SEM's, TEM's, EDS, LM's, Evaporators, Sputtercoaters, Recirculators, Ultra Microtomes, Enlargers and dark room equipment, Histology equipment, parts for some instruments and general lab equipment. SEM-TEM-Histology Services and sample preparation. e-mail your address and fax number for an Equipment List. Microscopy Labs Box 338 Red Bank, NJ 07701 fax 908 758 9142
I would like to obtain a PCVISIONplus framegrabber (PAL version). These boards made by Imaging Technologies Inc. were very popular some (6 to 8) years ago for monochrome image grabbing.
If you have one of these which you are not using and would be willing to part with it, would you please email me.
With thanks
David Vowles Electron Microscopy Unit Australian National University PO Box 475 Canberra ACT 2601 Australia Tel:(616) 2493543 Fax:(616) 2494891 Email: David.Vowles-at-anu.edu.au
Specimen blocks are very soft and are not clear but white.
I freeze yeast cells using a propane jet, freeze substitute in acetone for 4 days at -85C, replace acetone with acetone/uranyl acetate(about 0,1%) for 16 h during warm up to -35 C and wash with acetone for 2 h. Embedding: 1 h 50% HM20, 1 h 100% HM20, 16 h pure HM20, 6 h pure HM20, polymerization in gelatine cups using the Leica racks with the Leica AFS (but I use slightly smaller gelatine cups which fit directly onto Leica holder without the "Edelmann tubes"). I use relatively long times since agitation of samples is a problem in the AFS due to space limitations.
I am forwarding a message from Tony King at VG Microtech, who manufacture the Polaron range of sputter coaters, in regard to the recent question about AU/PD coatings.
Allow me to assist with your questions:
} Two questions: } 1. When a gold coated specimen is viewed in SEM at ~10k or higher, a } fine structure of gold coating becomes visible. Are there optimal } conditions of sputter coating (sputter current, time, distance } target-specimen, argon pressure, other gas than argon) to minimise the } artefact?
Without question, all of the parameters you mention are important for the high quality production of thin film for SEM observation. These are dependant on the design of the sputter head and vary from one design to the next it is impossible to give you further data without knowledge of the unit you use.
I would suggest that if you can see the coating at magnifications as low as 10k then coating is too thick! Try ten times thinner.
} 2. What is Au-Pd target? I thought that it was just an alloy one, but } a supplier of the coater says that using the alloy target is not } enough, that a coater with simultaneous sputtering Au and Pd from two } different targets is required to produce continuous coating.
The idea behind the alloy target is that the Pd acts as a barrier to the gold conglommarating into large islands which in turn obscures ultrastructure.
As there is a correlation between the work function of the metal and the sputter rate, then ideally two sputter heads with independant ontrol is the best option for Au/Pd , however, this is seen as prohibitively expensive for EM use, especially when other target amterials will give better results in some instances.
Another point of interest is that with a well designed magnetron sputter coater such as the Polaron range SC7640 system, the grain size and evenness of the coat is such that there should be no evidence of the coating with a standard SEM. This is of course dependant on the parameters you have stated earlier and a suitably thin coat.
Platinum, when used with the SC7640 will give results suitable for FEG SEM.
I hope this is of assistance to you, I have no doubt it will trigger some interesting debate on the open pages!.
Best regards Tony
Tony King Product specialist VG Microtech/ Polaron range
Tel: +44 (0)1825 746251 Fax: +44 (0)1825 768343
E&OE
Best regards, Steven Slap ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
Thanks to all who responded to my request for turbo pump experiences. I received over a dozen replies and there were only 1 or 2 problems=20 and these were on early models of turbo pump systems. =20 Most common remarks were; "reliable, clean, and trouble free." "Seldom need service" was also a repeated experience with the TP's. =20 Also frequently mentioned was the benefit of a vacuum system without=20 water and associated problems of corrosion, leaks etc. =20 Only negatives associated with newer turbo pumped systems was the cost= =20 of replacement or repair; many cited the wisdom of remaining on=20 service contracts.=20 =20 Possible problems with vibration and longer pump down times were=20 mentioned in a few responses but even these users with DP experience=20 preferred turbo pumped vacuum systems. =20 Thank you again for responding. =20 =20 John Humenansky Braun Intertec Corp. 6875 Washington Ave. So. Minneapolis, MN 55439 612-942-4822 =20
Thanks to all who have responded to the call for lead speciation in smelter vs natural occuring products. The information has been forwarded to the correct person and they will make there decision soon because the final report is due in 5 weeks. Thanks again.
Clarissa Vierrether The Doe Run Co. cvierret-at-misn.com
does anyone have or know of published modulation transfer functions of photographic film, especially Kodak SO-163.
Philip -- Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 86 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www_scem.csb.ki.se/pages/philip.html
MISSOURI-ILLINOIS-KANSAS MICROBEAM ANALYSIS SOCIETY & CENTRAL STATES MICROSCOPY SOCIETY
Presentations will be in the Arthur Mag Conference Center (Mag Center) of Midwest Research Institute, Kansas City, MO
APRIL 25, 1997 ====================================================== PROGRAM
8:30-9:00 Registration, Vendor Display Setup in the Mag Center.
9:00-9:10 Welcome by Dr. William P. Duncan, Director, Midwest Research Institute.
9:10-9:25 Dr. Peter Moroz, Jr., G.S. Technologies, Kansas City, MO "Microstructure of Metal"
9:30-9:45 Harold McCormick, C-K Engineering Inc., St. Louis, MO "Wearever of Metals"
9:50-10:05 Garry Crabtree, TechSpec Inc., Raytown, MO "Material Response to Deep Cryogenic Tempering"
10:10-10:30 Morning Break-Refreshments from BAR ROMA (cash cart).
10:30-10:45 Dr. Ody Maningat, Midwest Grain Products, Inc., Atchison, KS "Wheat Starch and Wheat Gluten Research"
10:50-11:05 Dr. Diane Durham, KU Medical Center, Kansas City, KS "Regeneration of Sensory Hair Cells in the Avian Cochlea-SEM Analysis"
11:10-11:25 Dr. Peter Smith, KU Medical Center, Kansas City, KS "Light and Electron Microscopic Investigation of Nervous System Plasticity"
11:30-11:45 Dr. Amit Mukherjee, KU Medical Center, Kansas City, KS "Electron Microscopic Analysis of the Polymerization of FtsZ, an Essential Cell Division Protein of E-Coli"
12:00-1:15 BUFFET LUNCH Generously provided by HITACHI, catered by Nance's Deli and Catering. Buffet includes vegetable manicotti, garden salad, garlic bread, soft drinks, etc.
1:15-1:35 Business Meetings
1:35-1:55 Paul Benson, The Nelson-Atkins Museum of Art, Kansas City, MO "Scientific Technique as Applied to Art Objects"
2:00-2:15 Marv Hart, Century Lubricants, Kansas City, KS "Lubrication and Grease Technology"
2:20-2:35 Garth Kristoff, Allied Signal, Kansas City, MO "Overview of the Technical Transfer and Solvent Substitute for Electronic Cleaning"
2:40-3:00 Afternoon Break-Refreshments from BAR ROMA (cash cart)
3:00-3:30 Michael Saba, Digital Instruments, Eden Prairie, Minnesota. "Applications in Scanning Probe Microscopy"
3:35-3:50 John Wilson, Mo. Regional Criminalistic Lab, Kansas City, MO "Capabilities of Crime Scene Investigation"
****************************************************** Complimentary lunch will be provided for all attendees by HITACHI. However, we need to know how many will be attending the luncheon, prior to the meeting. Please phone or email one of the following:
Larry Irwin 913-268-9009 Dan Kremser 314-935-5605 dkremser-at-levee.wustl.edu
} The french company ALTO MEDIA is producing a series of short movies (3 '), } based on a 'travel' into the matter.
Now it can be told... not really standard fare for this list, but I thought you all might enjoy this story.
Along the same lines, back in 1989 or so, some of the special effects for one of the Star Trek movies were done with SEM images. I think this was the first one Shatner directed, and a special-effects house called Associates & Ferran out on Long Island sold him on the use of SEM imagery because of the depth of field. If any of you remember the "planet at the end of the universe" scene, that "planet" was really a chunk of crystalline material imaged in the SEM at very low magnification.
When you hear that a film cost $50 million to make, here's how it happens. They bought a Zeiss SEM and a PGT imaging system just for the movie! A programmer who worked on camera motion-control animation systems was brought in to write stage control software to "fly" the stage through a sequence of images, which were stored on tape for post-processing. At that time, a 60MB cartridge tape was big storage. An overnight run filled up a tape and generated all of 3 seconds of animation! They did this more or less continuously for several months, and wound up using less than 10 seconds of it in the final film.
Some of the PGT engineers went to the movie the first week it opened. We got the very last credit line in the film, right behind the guys who bring the sandwiches for the actors' lunch. The people vacuuming popcorn off the theatre floor were giving funny looks to this bunch of strange folks standing in the aisles cheering at the credits 10 minutes after the movie was over and everybody else had left...
I've received lots of requests for the Microwave Staining procedure that I use for thin sections. So..... I'm sending this procedure out as a general post to everyone.
Sources for the procedure:
I use the general microwave techniques of Gary Logan, Ann Dvorak and R.T. Giberson. But, for this technique, the following is my own empirical findings. ( :-) so remember me when you reference!)
Notes on the procedure: =====================
* Section Collection:__________________ -- I collect my samples on grids, wick off excess water, and then immediately hold the back of the grid up to a 75 watt light bulb to dry. This seems to keep the sections on better. This is helpful because microwaving has a little more than the normal tendency to lift sections.
* Grid placement in stain:________________ -- When staining , I submerge the grid. Microwaving a floating grid tends to deposit uranyl acetate crystals on the grid. This is avoided by submerging the grid in the stain.
* Equipment:__________________________ --- I currently use a Ted Pella microwave, model 3440, but I've used a standard kitchen microwave before with good results.
--- Porcelain shallow well evaporating dish ( 12 wells)
--- Syringes &Filters for the stains used.
*Stains:_______________________________ --- Saturated (10%) Uranyl Acetate ( aq), less than 10 days old. Make in very small quantities. ( Our water , building or something is odd, this is only how long our aqueous solution lasts)
--- Venable's lead stain (John Venable, Richard Coggeshall,"A Simplified Lead Citrate Stain."The Journal of Cell Biology, 1965, Vol 25, pp407-408)
========================================= Procedure: ** Microwaving is actually only in the Uranyl acetate step.
1. For no more than 8 grids at a time, place about .25ml of filtered U.A. in a well for each pair ( identical pair) of grids, for a max of 4 wells that have U.A.
2. Fill all other wells with .25-.5 ml of water.
3. Place the grids , section side down, into and to the bottom of the drop. Be sure grids do not overlap.
4. Prewarm microwave that has 2 --300 ml water baths.
( Each Microwave oven is different, may need to test where best placement is. I use one water bath at 9 o'clock and one at 12 o'clock. Test using fresh cool water in the 2 beakers, place a liquid crystal sheet {Ted Pella} on the bottom, use a temp range of 35-40 or 40-45C......... look for the non-heated spots , this is where to place your sample)
5. Replace water in the water baths.
6. Place the evaporation plate in the "cool" spot of the microwave.
7. Microwave for 33 seconds and let plate set there for 6-8 minutes. Change water baths.
8. Repeat step 7 for a total of 4 times The total incubation time should be at least 20 min, but less than 30.
9. Leave the grids in the stain as you wash them one by one in warm water.
10. Proceed with normal lead staining. ( I use the Venable's for 40 seconds - Room temp).
FAQ's: ~~~ WHY MICROWAVE IF IT MAY TAKE 20 MINUTES ANYWAY?
The improvement in staining is really obvious. Also, I use a lower contrasting resin, and so this helps a lot. For things you are having trouble staining, this helps a lot.
~~~ CAN ONE OVER STAIN???~~~~~~~~~~~~~~~~~~~~~~~~~~~
Yes! Do a pilot study first. If you are working with a resin mixture that stains well, you may find that only 6-7 minutes is more than enough. I had a mixture at one time that did very well at 5 minutes only in total incubation time. I use what I do now for the way the mixture cuts.
~~~ WHY NOT JUST USE ONE BIG LONG MICROWAVE SESSION?
--The stain gets too hot, it will evaporate, and causes precipitation in the wells and on the grid. -- The sections will tend to come off if times of 1-2 min of microwaving are used. This is actually the method I started out with, and it doesn't work as well.
~~~WHAT ARE THE MOST COMMON PROBLEMS TO WATCH OUT FOR?
-- Overheating , will dry out the stain, and put stain ppt on the grid. -- Lifting sections, some batches of epoxy stick better than others, helps to dry the section on the grid with a light bulb, and to gently lower the grid in section side down. -- Over staining......was a new possibility for me! -- Lou Ann Miller Center for Microscopy & Imaging College of Veterinary Medicine Dept. of Veterinary Biosciences University of Illinois Rm 1108 Basic Sciences Bld 2001 S Lincoln Ave. Urbana, Illinois 61802
Phone: 217-244-1566 email: lamiller-at-uiuc.edu
Center for Microscopy & Imaging Home page: http://www.cvm.uiuc.edu/MicImagLab/MicImagLab.html
Central States Microscopy Society: http://www.cvm.uiuc.edu/Homepages/LouAnnMiller/CSMS/csms.html
Personal Home Pages: http://www.cvm.uiuc.edu/Homepages/LouAnnMiller/LAM.html
MISSOURI-ILLINOIS-KANSAS MICROBEAM ANALYSIS SOCIETY & CENTRAL STATES MICROSCOPY SOCIETY
Presentations will be in the Arthur Mag Conference Center (Mag Center) of Midwest Research Institute, Kansas City, MO
APRIL 25, 1997 ====================================================== PROGRAM
8:30-9:00 Registration, Vendor Display Setup in the Mag Center.
9:00-9:10 Welcome by Dr. William P. Duncan, Director, Midwest Research Institute.
9:10-9:25 Dr. Peter Moroz, Jr., G.S. Technologies, Kansas City, MO "Microstructure of Metal"
9:30-9:45 Harold McCormick, C-K Engineering Inc., St. Louis, MO "Wearever of Metals"
9:50-10:05 Garry Crabtree, TechSpec Inc., Raytown, MO "Material Response to Deep Cryogenic Tempering"
10:10-10:30 Morning Break-Refreshments from BAR ROMA (cash cart).
10:30-10:45 Dr. Ody Maningat, Midwest Grain Products, Inc., Atchison, KS "Wheat Starch and Wheat Gluten Research"
10:50-11:05 Dr. Diane Durham, KU Medical Center, Kansas City, KS "Regeneration of Sensory Hair Cells in the Avian Cochlea-SEM Analysis"
11:10-11:25 Dr. Peter Smith, KU Medical Center, Kansas City, KS "Light and Electron Microscopic Investigation of Nervous System Plasticity"
11:30-11:45 Dr. Amit Mukherjee, KU Medical Center, Kansas City, KS "Electron Microscopic Analysis of the Polymerization of FtsZ, an Essential Cell Division Protein of E-Coli"
12:00-1:15 BUFFET LUNCH Generously provided by HITACHI, catered by Nance's Deli and Catering. Buffet includes vegetable manicotti, garden salad, garlic bread, soft drinks, etc.
1:15-1:35 Business Meetings
1:35-1:55 Paul Benson, The Nelson-Atkins Museum of Art, Kansas City, MO "Scientific Technique as Applied to Art Objects"
2:00-2:15 Marv Hart, Century Lubricants, Kansas City, KS "Lubrication and Grease Technology"
2:20-2:35 Garth Kristoff, Allied Signal, Kansas City, MO "Overview of the Technical Transfer and Solvent Substitute for Electronic Cleaning"
2:40-3:00 Afternoon Break-Refreshments from BAR ROMA (cash cart)
3:00-3:30 Michael Saba, Digital Instruments, Eden Prairie, Minnesota. "Applications in Scanning Probe Microscopy"
3:35-3:50 John Wilson, Mo. Regional Criminalistic Lab, Kansas City, MO "Capabilities of Crime Scene Investigation"
****************************************************** Complimentary lunch will be provided for all attendees by HITACHI. However, we need to know how many will be attending the luncheon, prior to the meeting. Please phone or email one of the following:
Larry Irwin 913-268-9009 Dan Kremser 314-935-5605 dkremser-at-levee.wustl.edu
I'm looking for carbon EM calibration grid from SPI model 411CG-AB but couldn't find any information about SPI.Does anyone knows they phone # or www address? Thanks.
**************************** * Maoxu Qian, Ph.D. * * Dept of MSE, box 352120 * * University of Washington * * mxq-at-u.washington.edu * * (206)543-1514(phone) * * (206)543-3100(fax) * ****************************
SUMMER I 1997 COURSE ANNOUNCEMENT - Transmission Electron Microscopy (BIO. 221-Section BA)
NASSAU COMMUNITY COLLEGE, Garden City, Long Island, New York
A five week, Summer Session I 1997 semester, course in Biological Transmission Electron Microscopy is being offered by the Biology Department of Nassau Community College. This is a 4 credit course offered four days per week (Monday through Thursday) between the hours of 8:00 am and NOON. Classes will begin on May 27 and end on June 26, 1997.
This is a "hands-on" course emphasizing biological specimen preparation, ultra-thin sectioning involving block trimming, glass knifemaking and operation of the ultramicrotomes (Sorvall MT-2B and LKB Ultrotome III), thick and ultra-thin section mounting and contrast staining (UA and Pb citrate), grid support films (formvar, carbon), student operation of the TEM (Hitachi HS-8, Philips EM 300) and production of electron micrographs through the process of black & white photography, and electron micrograph analysis. Students will work on a chosen sample(s) with the goal of producing a portfolio of high quality TEM photomicrographs of that sample(s).
The course is widely transferrable and the cost per credit is reasonable at $78 per credit.
More information about the Bio-Imaging Center at NCC, course descriptions and syllabi, and the humble beginnings of a student gallery of EM photomicrographs is available at our web site. The URL is {http://www.sunynassau.edu/webpages/biology/becks.htm} .
Interested individuals should register as soon as possible since the course is limited to a total enrollment of ten (10) students.
If you have further questions, you should e-mail me directly at the address below.
Stephen J. Beck Associate Professor Bio-Imaging Center/Electron Microscopy Department of Biology Nassau Community College Garden City, NY 11530 Voice Mail: (516) 572-7829 Email: {becks-at-sunynassau.edu} URL: {http://www.sunynassau.edu/webpages/biology/becks.htm}
} does anyone have or know of published modulation transfer functions } of photographic film, especially Kodak SO-163.
There has not been much published lately... I believe that the most recent data comes from K. H. Downing and D. A. Grano, "Analysis of photographic emulsions for electron microscopy of two dimensional crystalline specimens," Ultramicroscopy, 7, 381-404 (1982).
-- Best Regards, John Minter
Eastman Kodak Company Phone: (716) 722-3407 Analytical Technology Division FAX: (716) 477-3029 Room 2112 Bldg 49 Kodak Park Site email: minter-at-kodak.com Rochester, NY 14562-3712 calendar: via PROFS
To all those who asked for a copy of a book chapter discussing staining: I am leaving for NYC for a week shortly. I will honor your request upon my return, April 22. Bye, Hildy
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Postdoc Position starting Oct. 1, 1997 at ANL-West
Argonne National Lab - West has an opening for a postdoc position to investigate irradiation damage in stainless steels from the EBR-II reactor. The project will mainly involve studying the defect structure and grain boundary segregation in up to ten different reactor core components. STEM will be the main technique used; however SEM, micro-hardness testing, and mechanical testing will be required occasionally.
Equipment available: JEOL 2010 STEM (LaB6) Zeiss 960A SEM Wide range of sample preparation equipment all capable of preparing radioactive samples.
Microscopy Skills Desired: Dark Field Imaging, EDS, Electron Diffraction, STEM, CBED
General Skills Desired: Experience handling/preparing radioactive samples Excellent verbal and written communication
Education Required: Ph.D. in Materials Science or Nuclear Eng. specializing in irradiation effects
This position will start Oct. 1, 1997.
Send Resumes to Brad Storey P.O. Box 2528 Idaho Falls, ID 83403
I would like to know what sort of probe current I should expect to measure at the specimen in a JEOL 2000fx TEM using a LaB6 filament, 200kV, 120 micron condenser aperture and spot sizes 1L to 6L. Under these conditions I measured the currents (using the faraday cup in our Gatan analytical specimen holder) to be:
1L 170nA 2L 66nA 3L 40nA 4L 7nA 5L 1nA 6L 0.3nA
Are these values reasonable? Cheers,
Mark Blackford TEM Group Materials Division, ANSTO PMB 1, Menai, N.S.W. Australia 2234 Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
can anyone tell me what purity of SF6 is required for use in TEM HT tanks and guns. I have a JEOL 2000 fx which was converted to SF6 in late 1994. We operate this machine at 200kV almost exclusively. At the time of the modification there was only one grade of SF6 available in Australia, as far as I could determine. I believe this was 99.8% pure. It has recently come to my attention that this may not be good enough. Before I go to the trouble and expense of sourcing higher purity SF6 I would like a definitive answer to the the question "how pure is pure enough?". JEOL Australia hasn't been able to help.
I look forward to any insight you can provide. Please respond to the listserver as there are several others in Australia that are keenly interested in this as well. Thanks,
Mark Blackford TEM Group Materials Division, ANSTO PMB 1, Menai, N.S.W. Australia 2234 Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
We have a JEOL 2010 and we use 99.995% purity SF6. I order it from a company which calls it VLSI grade , but I seem to recall that they used to call it Instrument grade.
We order a C-size cylinder (10 lb). Last time I ordered was about 2 years ago and we still have the cylinder with a fair amount of gas in it.
My background is in EM of biological, medical and food samples. I now have the opportunity to expand my skills and work on a set of materials samples.
My question is:
What are the standard methods for assessing surface degradation of materials samples (especially plastic polymers)? Are there macroscopic as well as microscopic methods which can give statistically "good" descriptions of the extend and type of degradation of such surfaces?
Thanks for any help (methods, references, review papers, etc.) you can provide. Please contact me offline.
Paula.
Paula Allan-Wojtas Food Microstructure Specialist Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia,Canada B4N 1J5
} For Sale: JEOL JSM T-200 scanning electron microscope. Purchased in } 1984; used about 50 hours, total time. Includes chiller and a small } sputter coater. $1500 OBO. Contact Dr. Jon Martin at 912/752-4060. } Located at Mercer University School of Medicine, Macon Georgia.
E-mail contact: HORST_MN-at-Mercer.EDU ******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
From the first superconducting magnet to the next generation of semiconductor chips, Oxford Instruments have always had an eye to the future - a future of scientific, human and commercial progress through the development of technology and people.
The Microanalysis Group is a highly successful international business within Oxford Instruments plc. We develop and manufacture high quality instrumentation for X-ray microanalysis and imaging of materials in electron microscopes. We are a world market and technology leader certified to BS EN ISO9001.
Continued expansion has led to an immediate vacancy in the Software R&D department for a researcher to improve our understanding of the physics of excitation & detection and develop innovative software algorithms to extend the range of microanalysis applications.
The successful candidate will have: - A relevant science degree or PhD with a good background in numerical methods and basic statistical theory. - Experience of EDX and/or WDX microanalysis and SEM theory and operation. - Programming skills in C and possibly Visual Basic on a PC platform.
In return for your commitment, we offer the full benefits package you would expect form a large plc. Career prospects are excellent and training will be tailored to the needs of the successful candidate.
If you are interested, please send a full covering letter of application and a comprehensive CV, quoting reference MRD01 on the envelope to:
Helen Bacon Personnel Manager Oxford Instruments Microanalysis Group Halifax Road High Wycombe, Bucks. HP12 3SE ENGLAND
or e-mail: helen.bacon-at-oxinst.co.uk -- Oxford Instruments MAG Software R&D
The Advanced International Immunofluorescence Course is a post-doctorate theoretical/practical course, with propedeutical lectures and practical stages on traditional and confocal immunofluorescence microscopy and image and ion analysis. The course will take place in Gargnano (Lake of Garda) from 7 to 10 October 1997. Further information and registration details will be found at the following Web address
http://imiucca.csi.unimi.it/endomi/ACIF.html
Thank you Paolo Castano _______________________________________________________________________
Prof. Paolo Castano UNIVERSITY OF MILAN INSTITUTE OF HUMAN ANATOMY - CHAIR OF HUMAN ANATOMY FOR PHARMACY Via Mangiagalli, 31 - 20133 Milan (Italy)
Dear fellow microscopists, Would anyone be interested in helping with a project on TEM at a high school level? My daughter Kelly, is doing research paper for intergrated sciences and is asked to find help from experts in the field (Mom's don't count) who can mentor her regarding some of the basic principles involved in TEM. She must cover chemistry, physics, mechanics and biology in relation to her topic. Her paper (due mid May, but preparing now) will cover some of these questions:
Why electrons are a better source than light for resolution? How does wavelength effect resolution? What makes Tungsten a good electron source, and how are the electrons generated? How does a a magnetic lens work? Why is a vacuum needed for operation? If electrons are invisible, how is the image generated off of the screen? How are images made on the film and then chemically developed? How does the eye focus and transmit this information to the brain?
If anyone would like to take a crack at any of the above with an explaination aimed toward the H. S. level (16yrs old. but fairly advanced science knowledge), it would be most appreciated. Of course, your mentoring would be acknowledged in her references. Please send all e-mail regarding this off line to me and I will forward if to Kelly. Thanks for promoting EM to young scientists! Linda Fox lfox1-at-wpo.it.luc.edu
It does seem strange that JEOL doesn't know what purity SF6 it prefers for its tanks. One of the other microscope vendors specified 99.9% or better in an installation guide for one of our TEM's.
} can anyone tell me what purity of SF6 is required for use in TEM HT tanks } and guns. I have a JEOL 2000 fx which was converted to SF6 in late 1994. } JEOL Australia hasn't been able to help.
Russell E. Cook Scientific Associate Electron Microscopy Center for Materials Research Argonne National Laboratory Building 212 9700 South Cass Avenue Argonne, IL 60439 (630)252-7194 FAX: (630)252-4798
The New England Society for Microscopy (NESM) announces its 14th ANNUAL SPRING SYMPOSIUM to be held at the Marine Biological Laboratories in Woods Hole, Massachusetts on May 9 & 10, 1997. Cohosted by the Connecticut Microscopy Society (CMS), the Metropolitan Microscopy Society (MMS) and the New York Society for Experimental Microscopists (NYSEM).
PROGRAM Friday, May 9th
12:00 pm Registration: Swope Center
1:00 pm Welcome: Lillie Auditorium
Session I Chairperson: Philip Leopold, NYSEM
1:10 pm "System Integration in Light Microscopy" Kenneth Spring, National Heart, Lung and Blood Institute, Bethesda, Maryland
1:50 pm "Diagnosis of Strange and Exotic Animal Diseases" Douglas Gregg, Foreign Animal Disease Diagnostic Lab, NVSL,USDA
2:30 pm "EM Site Magnetic Field Surveys and Solutions" Curt Dunnam, Cornell University, Linear Research Associates
3:10 pm Afternoon Break
Session II Chairperson: Joe Antol, CMS
3:30 pm A brief talk on "Amine Catalysts and Embedding Media" Jose Mascorro, Tulane University, MSA/LAS Director
3:50 pm "mRNA Localization and Cellular Morphogenesis" Gary Bassell, Dept. of Anatomy, Albert Einstein Medical School
4:30 pm "Quantitative Determination of Elemental Segregation at Interfaces in Solids" Tony Garratt-Reed, Center for Materials Science and Engineering, M.I.T.
5:30 pm Cocktails and Dinner: Swope Center
7:30 pm "Tracking the Giant Bluefin Tuna in New England Waters" Molly Lutcavage, New England Aquarium Edgerton Research Laboratory
Saturday, May 10th 7 to 8:00 am Breakfast: Swope Center
Session III Chairperson: Philip Flaitz, MMS
8:30 am "Ion Beam Milling Materials with Applications to TEM Specimen Preparation" Ron Anderson, IBM Analytical Services Group
9:10 am "Interaction of Viable Listeria Monocytogenes with Host Cell MHC Class II Compartments and Low pH Compartments" Paul Webster, Center for Cell Imaging and Department of Cell Biology, Yale School of Medicine
10:00 am Commercial Exhibits and Posters: Swope Center
12:30 pm Presentation of Poster and Photos-As-Art Awards, Door Prizes and "Nobska Light" Art Raffle Drawing: Poster Area, Swope Center
1:00 pm Lunch with Short Tour of MBL: Swope Center
2:00 pm 90-minute Discovery Cruise aboard the R/V Patriot II Tickets must be reserved in advance
This program is supported in part by a grant-in-aid from the Microscopy Society of America.
TO REGISTER, contact L. Kirstein at 104365.3522-at-compuserve.com. Advance registration is required. Deadline for cocktail/dinner reservations is April 25, 1997.
Evex Analytical is offering an X-ray detector enginer in its Princeton, New Jersey office.
Please foward resume to
Human Rresources Evex Analytical 857 State Road Princeton, NJ 08540
Candidates should have: - Experience of EDX and/or WDX microanalysis - A good background in numerical methods and basic statistical theory. - Programming skills in C, Visual Basic, Visual C++, Java, Active X
Please foward resume to
Evex Analytical 857 State Road Princceton, NJ 08540
The easiest, and most reliable method is to } buy commercially titrated NaOH (1 N ). No worries then. No need to try } a pH meter. Perfectly reliable.
This thread has offered some thought provoking ideas on the preparation of Reynold's. I have one question: how stable is commercially titrated NaOH? Doesn't a solution of NaOH absorb CO2 from the atmosphere and form sodium carbonate? Wouldn't this lead to a change in molarity with time and promote the formation of lead carbonate ppt on the sections?
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
} can anyone tell me what purity of SF6 is required for use in TEM HT tanks } and guns.
We have a JEOL 4000, which uses Commercial Grade SF6, and an HVEM, in which we have traditionally used Instrument Grade SF6. Due to the re- cent price explosion for SF6, we have been investigating whether we can use the Commercial Grade SF6 in the HVEM as well. The major difference between the two grades is that there is more N2 and O2 in the Commercial Grade. The HVEM has a gas storage facility with several filters and a molecular seive through which SF6 can be cycled. I think that much of the N2 and O2 can be eliminated by venting a small amount of SF6 to the air before putting the rest in the tank. You'd have to calculate the dielectric constant and know the relevant spark gap widths, etc. to be sure if Commercial grade would work for you, but our conclusion at this point is that it works for the 400 kV instrument, and should be OK for the HVEM as well. (We are still going to analyse both grades for im- purities.) Good luck. Yours, Bill Tivol
} What are the standard methods for assessing surface degradation of } materials samples (especially plastic polymers)? Are there macroscopic } as well as microscopic methods which can give statistically "good" } descriptions of the extend and type of degradation of such surfaces? } Dear Paula, I recently ran across a good review "Electron Microscopy in Polymer Science" by G.H. Michler, Applied Spectroscopy Reviews, vol 28(4), pp 327-384 (1993). This paper discusses surface characteriza- tion and many other topics. Additionally, I suggest STM or AFM as possibilities, but I don't have any referrences for them. Perhaps comparing the specular vs diffuse reflectivities of polymer surfaces before and after damage would be informative for degradation features ~ 1 micrometer in size. Good luck. Yours, Bill Tivol
Just a quick note to let you all know that that Microscopy Listserver Archives are now on-line. I have made accessible all Email postings covering the period Oct.1993 through Mar.1997 and will update the archive monthly.
The index is not directly searchable, however, it is chronologically sorted by Month and Year.
If you download a given month's postings then you can search the downloaded WWW page for any keyword/phrase that you wish by using the native search/find option of your WWW Browser. (In NetScape this is located in the Edit Pull Down Menu and is called FIND).
You may access the archive at the MSA WWW site.
http://www.msa.microscopy.com
just follow the links to the Reference/Educational Activities Page.
I'll get around to putting together a completely searchable index sometime in the forseeable future, but for now this will go a reasonable way to letting everyone find "old messages and postings" and all the other miscellaneous requests I receive for information.
Way back in the mid 1980's there was a book called "Thin Is In" and it dealt with staining of samples embedded in GMA. Are there still copies out there? Or can someone direct me to where I can get ahold of the book so I can phototcopy pertinent pages? I'm sorry, but I don't remember who the author was.
Thanks in advance for all you help.
Paula = )
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
A while ago I remember reading that not only should the lead stain be made up at the proper pH (12), but both the lead citrate powder (for Venable's) and the NaOH need to be fresh. I bought new reagents (esp. C02-free NaOH solution) and got much cleaner, stronger staining.
Also, I never had very great uranyl acetate staining. When I was doing work on collagen, I switched to a tannic acid/uranyl acetate stain (ref: Kajikawa et al., 1975, J. Electron Microsc. 24:287-289) and I have since used it on most everything. Again, it seems to give stronger, cleaner staining than the UA alone. This stain (a modification of the one in the reference) must be made fresh on the day of use, so I usually make fairly small volumes. I pre-weigh 0.04 gm tannic acid into a few tubes. On day of use add 6.8 ml distilled water to one tube, mix and hold in hands or place in oven about 5 minutes to warm. Mix again, then add 200 ul of 2 % aqueous uranyl acetate. Mix and spin down or put through syringe filter before use. Stain 10-15 minutes and rinse in water before going to the lead stain.
Another reason I like the above solution is that, according to my calculations anyway, if I start with depleted uranyl acetate to make the 2%, the final tannic acid/UA stain solution does not contain enough radioactivity to be considered radioactive. So I don't have to worry about radioactive staining dishes, forceps, etc. etc. as long as I am careful with the original UA and 2% solutions. Of course most of the materials I use are disposable, and the waste profiles are designated to contain some UA so it really doesn't matter alot. But it makes me feel better to limit usage of the stuff.
Now if I could just learn how to stain sections on formvar-coated grids without getting lots of folds and dense pockets!
Good luck,
Karen Zaruba
} This should be a simple procedure but I am having a terrible time trying to } stain some Embed 812 embedded tissues with uranyl acetate/lead citrate. The } tissue is from frog tadpoles, fixed in glutaraldehye/paraformaldehyde and } postfixed in osmium. I have tried staining for 5-20 minutes with saturated } solution of UA in ethanol, followed by 1-5 minutes in lead citrate } (Venable-Coggeshall formulation). The sections look no different from } unstained specimens. } } I'd appreciate any suggestions about what I might be doing wrong. } } } Gary Radice, Associate Professor gradice-at-richmond.edu } Department of Biology 804-289-8107 (voice) } University of Richmond VA 23173 804-289-8233 (FAX) }
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Life Sciences Sector Lab Reply: kszaruba-at-mmm.com 3M Company 3M Center 270-1S-01 Phone: 612-737-2971 St. Paul, MN 55144-1000 Fax: 736-1519
These opinions are my own and may not represent those of 3M.
I have recently been asked to take over an ICP. Since this newsgroup has been so helpful with microscopy issues I was wondering if anyone knows of a similar newsgroup dedicated to ICP spectroscopy.
Could someone please give me the name of a book which would have TEM pictures of paper which has been prepared with ultramicrotomy. Or if they have one or two thay would be willing to send to me privately I would really appreciate it.
For our JEOL 2010, Jeol specify a purity of 99.99% or better in their maintenance manual. We however have 99.995% purity obtained from Scott Specialty Gases, Fremont.
Best Regards,
Keith. ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Dr. K. Moulding.
Materials Characterisation and Preparation Facility Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong.
Below is a request for instructions and/or parts for a cryostat from some colleagues.
Thanks in advance for your help!
Tina **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* **************************************************************************** Message:
We have recently obtained a surplus cryostat, a Milles Scientific Microtome Model 4553. The unit chills well and the microtome is mechanically in good shape. Unfortunately, it does not have instructions, an antirolling plate or specimen stubs. Does anyone know a source for these items? We have been unable to locate a phone number or address for Milles Scientific. Perhaps they have gone out of business or merged with another company. Perhaps someone has surplus parts for this model we could acquire. Your assistance would be appreciated.
In nearly 30 years of preparing lead citrate according to Reynolds I have experienced no problems in using a stock sodium hydroxide solution (1M) which was made up from pellets. However, this solution must be freshly made up. Is it because I always use Reynolds in its concentrated form that I do not experience any problems?
Rob
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa eurc-at-giraffe.ru.ac.za - tel: +27 461 318168 - fax: +27 461 24377
Materials Analysis Group, Philips Semiconductors, has an immediate opening for an experienced XRD analyst.
Responsibilities include operating XRD and scanning acoustic microscopy on bulk/thin film specimens and packaged devices, respectively, primarily for external customers on a commercial basis. Analytical support work for internal customers on silicon IC production issues will also be required.
Qualified candidates must possess a Ph.D. or M.S. degree in a field such as Materials Science or have equivalent expertise. They must have extensive hands-on experience in operating XRD equipment and in providing analyses to customers, preferably as a member of an analytical services laboratory. Familiarity with IC processing would also be advantageous.
Equipment available includes a Siemens D500 with grazing incidence and thin film reflectivity attachments, and a Sonoscan C-SAM. Acquisition of a thin film diffractometer is under consideration. The laboratory is also well equipped for SIMS, Auger, ESCA, TEM, SEM AFM, FIB, Raman, FTIR, etc.
Located in the heart of Silicon Valley, 40 miles south of San Francisco, Philips Semiconductors offers a generous benefits package that includes life, health and dental insurance, a pension plan, and a company-matched savings and investment plan.
For confidential consideration, send your resume to: Alan Morgan, Materials Analysis Group, Philips Semiconductors MS 65, 811 E. Arques Avenue, Sunnyvale, CA 94088: FAX (408)991-4801.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Paula, Thin Is In: Plastic Embedding of Tissue for Light Microscopy. Burns, William A. & Bretschneider Ann. 1981. Educational Products Division. American Society of Clinical Pathologists. Chicago. ISBN 0-89189-083-1
I have a copy, so send your fax number, what you would like copied I'll get them to you. Ian.
Fellow microscopists I am doing some TEM of cultured lymphocytes. All membranes seem to be absent. There are halos where membranes should be but no membranes. So far I have tried 2.5 glut in 0.1M cacodylate 2.5 glut in 0.2M cacodylate (caused shrinkage) 4 paraformaldehyde 1 glut in 0.1M cacodylate.
Post fixing in Osmium dehydrating in ethanol and embedding in Spurr
I am going to try making up the fix in the culture medium next.
Has anyone any thoughts of anything else to try. I would be particularly interested in the use of Ca+ Mg+ and sucrose.
Many thanks
Chris Chris Gilpin Biological Sciences Electron Microscope Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 161 275 5170 fax +44 161 275 5171
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America Like Rob I've been making Reynold's lead citrate since I was a boy. I use NaOH pellets but degas the distilled water by sonicating it for a few minutes before making the stain.
A discussion of SF6 took place on this list back in December. At that time I received an off-line message from John Giles about Dilo Company. Dilo sells (and rents) a system that reclaims the SF6 in your tank, purifies it and pumps it into storage tanks for reuse. Dilo's business is primarily with big consumers of SF6 - power companies, but they are knowledgeable about SF6 purity and applications. What interested me was they sell a small system for ~$5,000, which is what a tank of SF6 costs in some places (like the U.P. of Michigan).
I have not used their system. Has anyone else?
You can reach them at;
Dilo Company, Inc. 231A Douglas Rd.., Unit 5 Oldsmar, FL 34677 813-855-1448 http://www.dilo.com dilo-at-cent.com
I have no interest in Dilo Company other than as a consumer. Owen P. Mills Michigan Technological University Metallurgical & Materials Engineering Rm 512 MME Building Houghton, MI 49931 906-487-2002 906-487-2934 FAX opmills-at-mtu.edu
At 03:49 PM 4/15/97 -0700, Jill Craig asked: } I have recently been asked to take over an ICP. Since this newsgroup } has been so helpful with microscopy issues I was wondering if anyone } knows of a similar newsgroup dedicated to ICP spectroscopy. } I have used Netscape search engines with no luck.
I don't know the specific answer, but the best way to find lists such as this is to use http://www.liszt.com
Best regards, Steven Slap ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
Is anyone out there looking for a used Duo Mill for materials TEM sample prep? Excellent working condition and a very reasonable price. If interested contact Kim Jones 303-421-3182 or send e-mail kjonesVMS-at-aol.com
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Chris, For cultured cells I routinely fix in 2% glut/0.1M. cacod and post fix with 1% OsO4/0.1M. cacod, embed in Araldite, then stain sections with Ua/Pb with no problems. What are your fixation conditions - monolayer, pellet of cells. From your description sounds like the OsO4/Pb side of things is at fault. I wouldn't add fixatives to the culture medium, depending on its constituents you might have a Canazarro type reaction and fix the medium as well as the cells. Ian.
Me, too. I boil my water, cool it a little, dissolve the pellets and make the whole mess warm. I use the fancy carbonate-free pellets. I don't keep the NaOH, either-just add it to my buffer-adjusting stock. I've never tried sonicating but sounds good and easy. I wouldn't dream of sticking my pH electrode in my lead-I use a fresh piece of close-reading paper. Seems to work just fine. Grace
Been using 99.8 SF6 for several years in our JEOL 2010. Generally works well. We have groups working at 100 & 200KV. When not used at 200kv for several mo., some dark current instability is exibited when returning to 200KV. With a bit of patients we are back in business. The stability problem ceases to exist with regular use at 200KV.
I've noticed a lot of people seem to be using Reynold's Pb stain. In the past (20 years ago) I used Reynold's also. There seemed to be too many problems with the staining. I started using lead citrate to make up my stain. Consistantly good results. Here's how if anybody is interested.
Distilled water in clear glass storage bottle...........about 80 ml lead citrate.................................................0.4gm
Stir vigorously on magnetic stirrer for several minutes avoiding bubbles.
10N NaOH (I buy from Fisher already made up. 100 ml bottle).....0.2ml
Continue to stir until solution is clear. Bring up to 100ml. Store in 4C refrigrator. Good for up to 6 months. Avoid shaking it.
Stain grids for 4 minutes, rinse grids.
As far as flat beer goes, try adding CO2 under vacuum and let it sit for a day.
Peace through Christ,
Phil Rutledge, Director e-mail: prutle1-at-gl.umbc.edu Center for Microscopy voice: (410) 455-3582 UMBC Dept. of Biology fax: (410) 455-3875 Catonsville, MD 21228 ///// / / / / /////// /////// / / /////// /////// / / / / / / / / / / / / /////
Would freshly distilled water be equivalent to degassing via sonication ?
Leo
On Wed, 16 Apr 1997, Ian Montgomery wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } -----------------------------------------------------------------------. } } } } Good morning Reynolds users } } } } In nearly 30 years of preparing lead citrate according to } } Reynolds I have experienced no problems in using a stock } } sodium hydroxide solution (1M) which was made up from pellets. } } However, this solution must be freshly made up. Is it because I } } always use Reynolds in its concentrated form that I do not } } experience any problems? } } } } Rob } } } Like Rob I've been making Reynold's lead citrate since I was a boy. } I use NaOH pellets but degas the distilled water by sonicating it for a few } minutes before making the stain. } } Ian. } } }
To All I have been doing TEM since 1970, and have always used NaOH pellets (CP grade - which do contain about 0.5 - 1% Na2CO3). I make up a 10N solution in small aliquots (~20 ml), and dump it when I see crystals of sodium silicate (NaOH dissolves glass). I use this 10N solution to add to the powdered Pb citrate in dH2O. I make up Pb citrate in 100 ml quantities and dump it when it turns cloudy. Usually it keeps for months, unless someone forgets to tighten the cap, which is not uncommon in a central service lab. What I have learned in nearly 30 years of TEM, and many years in other microscopy techniques is that there are very few empirical rules. I have been in very fussy labs that used aged Pb citrate in bottles that were so coated with precipitate that they looked out of a sunken pirate ship, and have been in other labs that make up Pb citrate fresh, and filter it. Whatever works. Bruce Cutler, Microscopy Lab, Univ. Kansas, Lawrence KS
I am looking for information and possibly a handbook on an American optical Micro-Star Illuminator model 1872 (I am new to this scope). This microscope was discontinued and replaced by another microscope - the EpiStar metallurgical microscope which (I believe) has been discontinued itself.
I would like to hear from anyone who has used this scope. I am also in need of objectives (the optical kind).
Chris: What are the times and temperatures? Tissue cultured cells require only about 5 minutes in each fixative at 20 degrees C or at the most for 15 minutes in ice, unless you use much lower concentrations. Overfixing or storage in con. ethanol removes lipids and hence membranes. Badly Os overfixed tissues show dark cytoplasms surrounded by "white" cell membranes. Regards Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 300+ Links, MSDS ************************ http://www.proscitech.com.au
} I am doing some TEM of cultured lymphocytes. All membranes seem to } be absent. There are halos where membranes should be but no } membranes. } So far I have tried } 2.5 glut in 0.1M cacodylate } 2.5 glut in 0.2M cacodylate (caused shrinkage) } 4 paraformaldehyde 1 glut in 0.1M cacodylate. } } Post fixing in Osmium dehydrating in ethanol and embedding in } Spurr } } } I am going to try making up the fix in the culture medium next. } } Has anyone any thoughts of anything else to try. I would be } particularly interested in the use of Ca+ Mg+ and sucrose. } } } Many thanks } } Chris } Chris Gilpin } Biological Sciences Electron Microscope Unit } G452 Stopford Building } Oxford Road } Manchester } M13 9PT } phone +44 161 275 5170 } fax +44 161 275 5171
I and other collegues have been trying fruitlessly to e-mail Kevex and when I tried to find their web page it was not there either. Is this just a faulty computer or have they been restructured out of existence? Does anyone out there know?
No, freshly distilled water is not necessarily degassed. Air can redissolve nicely as the water drips down from the condenser into the collecting flask.
But as an alternative to sonification, you can try just plain BOILING. This reduces the amount of dissolved gas significantly. Then just let it sit. Don't shake it with air, try not to pour it too much. This worked for us with a different degassing problem a couple years ago.
Cindy Bennett Hoechst Diafoil GmbH Wiesbaden, Germany bennett-at-msmhdg.hoechst.com ---------- Von: Leo Marin An: Ian Montgomery Cc: Microscopy-at-Sparc5.Microscopy.Com Betreff: Re: NO NO NO pellets for Pb stain Datum: Donnerstag, 17. April 1997 01:10
----------------------------------------------------------------------- - The Microscopy ListServer -- Sponsor: The Microscopy Society of America -----------------------------------------------------------------------
} To All } I have been doing TEM since 1970, and have always used NaOH pellets } (CP grade - which do contain about 0.5 - 1% Na2CO3). I make up a 10N } solution in small aliquots (~20 ml), and dump it when I see crystals } of sodium silicate (NaOH dissolves glass). I use this 10N solution } to add to the powdered Pb citrate in dH2O. I make up Pb citrate in } 100 ml quantities and dump it when it turns cloudy. Usually it keeps } for months, unless someone forgets to tighten the cap, which is not } uncommon in a central service lab. What I have learned in nearly 30 } years of TEM, and many years in other microscopy techniques is that } there are very few empirical rules. I have been in very fussy labs } that used aged Pb citrate in bottles that were so coated with } precipitate that they looked out of a sunken pirate ship, and have } been in other labs that make up Pb citrate fresh, and filter it. } Whatever works. } Bruce Cutler, Microscopy Lab, Univ. Kansas, Lawrence KS
I store our freshly made-up concentrated Reynolds solution in 3ml aliquots in Eppendorff tubes at 4C. This way there is minimal exposure to CO2 and no danger of someone leaving the container open. A tube of stain is only used once, anything left over in the tube being discarded. The stain keeps well for months in these tubes.
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa eurc-at-giraffe.ru.ac.za - tel: +27 461 318168 - fax: +27 461 24377
Dear colleagues, Does anybody know why everybody uses for section constrasting uranyl acetate which solubility is not so high but not uranyl nitrate or uranyl sulfate which are much more soluble?
Sincerely yours, Alexander Mironov
Consorzio Mario Negri Sud S. Maria Imbaro (Chieti) Italy
Keith: I can't help you with sonicating water, but there is only one thing to do with flat beer, bin it. But then you people in the West Country have no idea what constitutes good beer. Come to Cambridge for a pint of Greene King Abbot or, better still Adnams. Both will knock your socks off
PatrickOn Wed, 16 Apr 1997, Keith Ryan wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Ian } } I really appreciate the tip about sonicating water to degas it, I would not } have believed it. Now, do you have any suggestions for flat beer? } } Regards - Keith Ryan } Plymouth Marine Lab., UK } } }
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America If my memory serves I got sonication from the Technical Hints and Tips in the Proceedings of the RMS. I cover my options by sonicating the water whether fresh or hours, days old. Ian.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America If my memory serves I got sonication from the Technical Hints and Tips in the Proceedings of the RMS. I cover my options by sonicating the water whether fresh or hours, days old. Ian.
The discussion about UALC staining is wonderful. It's interesting that what works for one doesn't somewhere else.
We went back to Reynold's after problems with the Venable & Coggeshall version. But we store in in 20ml syringes in the refrigerator. It keeps a long time (6-12 months) with no exposure to the air. It filtered through a .22 µm syringe filter. We leave a needle on it and insert it into a rubber stopper. Before use we express a few drops and then place drop onto parafilm in a petri dish.
Good day,
Chuck ------------------------------------- Name: Charles Gilbert VOC:(704)355-5261 Carolinas Medical Center FAX:(704)355-8424 Dept of Pediatric Research PO Box 32861 Charlotte, NC 28232-2861
Chris,
We use 3% glut in 0.1M cacodylate for hematology samples. The membranes are not very well preserved. They are often blurred, partial and sometimes absent but nothing like what you have described. The cells still appear bounded.
We have used cryofixation-freeze substitution and have found excellent membrane preservation in cell suspensions right down to the trilaminar plasmalemma.
Next best is a dimethyl sulfoxide pretreatment before fixing in glut. We use 10% in RPMI-1640 culture media for 10 min. You could probably use whatever media your cells are growing in. This has given us better membrane boundaries than glut alone. They show up unilaminar.
Caveat -- DMSO has some interesting properties and effects on cells. It's best to study the literature especially the safety requirements. It's probably not good to just dump it down the sink.
Best wishes,
Chuck ------------------------------------- Name: Charles Gilbert VOC:(704)355-5261 Carolinas Medical Center FAX:(704)355-8424 Dept of Pediatric Research PO Box 32861 Charlotte, NC 28232-2861
could you please add me to your mailing list (Martin Roe)
In response to Mel Dickson's posting, and to reassure the many Kevex customers on this forum, Kevex is indeed alive and well, but we are experiencing a problem with our Internet service (web site and email) that was just discovered today. As a result, this morning we have been absolutely flooded with telephone calls and other messages reporting this problem. We are sorry for this inconvenience and we are working with our Internet service provider to restore our presence on the Internet as quickly as possible.
Alternate contact methods:
Kevex main office
Tel: +1 (800) 865-3839 (toll free in the USA) Tel: +1 (805) 295-0019 Fax: +1 (805) 295-0419
Kevex main service office
Tel: +1 (800) 495-3839 (toll free in the USA) Tel: +1 (415) 562-2500 Fax: +1 (415) 562-2505
For information on regional offices and other offices worldwide, please contact either of the offices listed above.
Thanks to all who responded to my query about a source for parts. I had thought that Leica was the place and since they are just down the road from us...
In response to Mel Dickson's posting, and to reassure the many Kevex customers on this forum, Kevex is indeed alive and well, but we are experiencing a problem with our Internet service (web site and email) that was just discovered today. As a result, this morning we have been absolutely flooded with telephone calls and other messages reporting this problem. We are sorry for this inconvenience and we are working with our Internet service provider to restore our presence on the Internet as quickly as possible.
Alternate contact methods:
Kevex main office
Tel: +1 (800) 865-3839 (toll free in the USA) Tel: +1 (805) 295-0019 Fax: +1 (805) 295-0419
Kevex main service office
Tel: +1 (800) 495-3839 (toll free in the USA) Tel: +1 (415) 562-2500 Fax: +1 (415) 562-2505
For information on regional offices and other offices worldwide, please contact either of the offices listed above.
I saw their site just a week ago, but am not able to see it today. Maybe something did happen administratively. Anyone else know?
At 01:37 PM 4/17/97 +1000, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The discussion about UALC staining is wonderful. It's interesting that what works for one doesn't somewhere else.
We went back to Reynold's after problems with the Venable & Coggeshall version. But we store in in 20ml syringes in the refrigerator. It keeps a long time (6-12 months) with no exposure to the air. It filtered through a .22 µm syringe filter. We leave a needle on it and insert it into a rubber stopper. Before use we express a few drops and then place drop onto parafilm in a petri dish.
Good day,
Chuck ------------------------------------- Name: Charles Gilbert VOC:(704)355-5261 Carolinas Medical Center FAX:(704)355-8424 Dept of Pediatric Research PO Box 32861 Charlotte, NC 28232-2861
Chris,
We use 3% glut in 0.1M cacodylate for hematology samples. The membranes are not very well preserved. They are often blurred, partial and sometimes absent but nothing like what you have described. The cells still appear bounded.
We have used cryofixation-freeze substitution and have found excellent membrane preservation in cell suspensions right down to the trilaminar plasmalemma.
Next best is a dimethyl sulfoxide pretreatment before fixing in glut. We use 10% in RPMI-1640 culture media for 10 min. You could probably use whatever media your cells are growing in. This has given us better membrane
boundaries than glut alone. They show up unilaminar.
Caveat -- DMSO has some interesting properties and effects on cells. It's best to study the literature especially the safety requirements. It's probably not good to just dump it down the sink.
Best wishes,
Chuck ------------------------------------- Name: Charles Gilbert VOC:(704)355-5261 Carolinas Medical Center FAX:(704)355-8424 Dept of Pediatric Research PO Box 32861 Charlotte, NC 28232-2861
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Reply to: RE} FWD} What has happened to Kevex?
Kevex has a 800 number for field service and messages to others. Try 1-800-495-3839
--------------------------------------
Hello world.
I and other collegues have been trying fruitlessly to e-mail Kevex and when I tried to find their web page it was not there either. Is this just a faulty computer or have they been restructured out of existence? Does anyone out there know?
How do you sonicate to remove gas bubbles? Do you immerse a probe, or place the container in an ultrasonic bath?
Thanks, Jim Williams Indiana University
} } Ultrasonication has been a standard technique for degassing fluids in our } labs for many years. (Before that we boiled where possible or pulled a } gentle vacuum on a closed container. Main purpose was to degas solutions } to be passed through automatic light blockage partcle counters. (Air } bubbles count very nicely as particles.) Nice thing about sonication is } that you can safely sonicate oils and other fluids that you wouldn't want } to boil or pull into a vacuum system. } } Bob Holthausen } Pall Corporation } } } } } } {snip} } If my memory serves I got sonication from the Technical Hints and } Tips in the Proceedings of the RMS. I cover my options by sonicating the } water whether fresh or hours, days old. } Ian.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Reply to: RE} FWD} What has happened to Kevex?
Kevex has a 800 number for field service and messages to others. Try 1-800-495-3839
--------------------------------------
Hello world.
I and other collegues have been trying fruitlessly to e-mail Kevex and when I tried to find their web page it was not there either. Is this just a faulty computer or have they been restructured out of existence? Does anyone out there know?
---------------------------------------------------------- Warning: AOL4FREE.COM is a Trojan Horse Program, which will erase your Hard Disk. ----------------------------------------------------------
Colleagues...
I have checked this announcement out on the U.S. DoE Computer Advisory WWW Page. It is for real so please take notice if you are a DOS/Windows User. Extended details can be found at the WWW address.
http://ciac.llnl.gov/
Technically the program is a Trojan Horse not a virus so it will NOT BE DETECTED by typical virus protection software.
Note, this is a different program from the MacIntosh AOL4FREE program which also circulated the net recently. That one created fraudulant accounts on AOL which is a different issue all together. It was illegal but did not damage your computer. This one will wipe out your disk. A few details are listed below but check out the WWW page at CIAC for the latest information.
A Trojan horse program called AOL4FREE.COM is circulating on the Internet. This is not a virus and cannot be detected by most anti-virus programs. The program is executed by the individual user and deletes all files on a hard drive.
PLATFORM: DOS/Windows-based PCs
DAMAGE: When the AOL4FREE.COM program is executed, all files and directories on the user's C: drive are deleted.
DO NOT execute this program. [Note: Double clicking on.com or .exe will start a selected program.] If the program starts executing, quickly pressing Ctrl-C will save some of your files. If this has happened, shut down immediately and call for assistance. DO NOT attempt to write anything to your hard drive. Files that have been destroyed may be able to be recovered IF you have not written anything to the hard drive and IF your system has not written anything such as when utilizing a auto-save program.
Please see CIAC Bulletin H-47 available on the CIAC homepage (http://ciac.llnl.gov/) for detailed information.
We use 3% glut in 0.1M cacodylate for hematology samples. The membranes are not very well preserved. They are often blurred, partial and sometimes absent but nothing like what you have described. The cells still appear bounded.
We have used cryofixation-freeze substitution and have found excellent membrane preservation in cell suspensions right down to the trilaminar plasmalemma.
Next best is a dimethyl sulfoxide pretreatment before fixing in glut. We use 10% in RPMI-1640 culture media for 10 min. You could probably use whatever media your cells are growing in. This has given us better membrane
boundaries than glut alone. They show up unilaminar.
Caveat -- DMSO has some interesting properties and effects on cells. It's best to study the literature especially the safety requirements. It's probably not good to just dump it down the sink.
Best wishes,
Chuck ------------------------------------- Name: Charles Gilbert VOC:(704)355-5261 Carolinas Medical Center FAX:(704)355-8424 Dept of Pediatric Research PO Box 32861 Charlotte, NC 28232-2861
Research Associate Position
"ELECTRON MICROSCOPY OF MATERIALS"
The Department of Metallurgy and Materials Science, University of Toronto plans to make a term appointment of up to 6 years duration in the area of electron microscopy of materials, with a starting date as early as 1 July 1997. The starting annual salary will be $40,000 plus fringe benefits. The ideal candidate will possess a Ph.D. in a relevant field with significant experience in electron microscopy-based research. The Department operates a facility with a conventional 200 kV TEM/STEM/EDX and 4 computer networked SEM instruments including a high-resolution FESEM with BSE/EDX/EBIC/OIM capabilities. Secondly, a coordinated McMaster University/University of Toronto high-resolution JEOL 200 kV FETEM/STEM with EDX/PEELS/HAADF facility is located at nearby McMaster University. In addition the University of Toronto supports other accessible electron microscopy equipment including a VG HB601 STEM with EDX/PEELS. The candidate will be expected to supervise various activities in the Departmental facility with the support of 3 technical staff members. It is expected that the candidate will be involved in various research projects in conjunction with university/industry users of the facility. The ideal candidate will have experience with a range of microscopy techniques to support ongoing research projects in such areas as semiconductor nanostructures, interfacial segregation analysis, shape memory alloys and biomaterials. Applications, including a curriculum vitae and three letters of reference should be sent to Professor D.D. Perovic, Director, Electron Microscopy Facility, Department of Metallurgy and Materials Science, University of Toronto, 184 College Street, Toronto M5S 3E4 Canada; Fax: (416) 978-4155, Email: perovic-at-ecf.utoronto.ca. The deadline for the receipt of applications is 1 June 1997
_________________ D.D. Perovic Department of Metallurgy and Materials Science, University of Toronto 184 College Street, Toronto M5S 3E4 Canada Tel: (416) 978-5635 Fax: (416) 978-4155
} } } Has anyone comments on the UV polimeriztion of Lowicryl HM20 after freeze } substitution in Acetone-Uranylacetate. I have problems with } prepolymerization of the resin which causes unusable blocks, but the } problem is not consistent, some blocks are good some are not. } } TIA, } Stefan }
I have some experience of this technique. Because HM20 is polymerised by UV light, it is essential that the UV can penetrate the specimen fully, therefore any heavy metals such as osmium or uranyl acetate can potentially cause problems by "shading" areas of the resin from proper polymerisation.
You can minimise the problem by only using mild concentrations of UA in the acetone, by making the blocks as small as possible, and perhaps rinse with pure acetone several times if your substitution apparatus allows it, before polymerising the blocks. Also extend polymerisation time, although this may affect antigenicity. Good luck!
Dare Chris I had the same problem once when I was processing some cultured cells for EM. Morphology was bad at all, but there was no contrast on all membrane. Then I processed the same cell culture with the same batch of osmium solution and fresh osmium side by side. It turned out fresh osmium solved problem.
I have DT2803 frame grabber from Data Translation but without any manual and software. From Data Translation I can't get any information, because this product is out of the production. Please help me. -- Henrik Kaker SEM-EDS Laboratory, Metal Ravne d.o.o., Slovenia Tel: +386-602-21-131, Fax: +386-602-20-436, E-mail: Henrik.Kaker-at-guest.arnes.si http://www2.arnes.si/guest/sgszmera1/index.html Microscopy Vendors DataBase: http://www.kaker.com/mvd/vendors.html Kaker.Com: http://www.kaker.com
Thanks to all of you who responded to the SEM for sale at Mercer University. The microscope has been sold. The folks there are grateful to this forum. ******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
} But as an alternative to sonification, you can try just plain BOILING.
Actually, heating to somewhat below boiling will allow the gas to escape. Another possibility is to attach a side-arm flask to the house vacuum and swirl the solution. That works for the case that the solution to be degassed cannot be heated. Yours, Bill Tivol
Ultrasonication has been a standard technique for degassing fluids in our labs for many years. (Before that we boiled where possible or pulled a gentle vacuum on a closed container. Main purpose was to degas solutions to be passed through automatic light blockage partcle counters. (Air bubbles count very nicely as particles.) Nice thing about sonication is that you can safely sonicate oils and other fluids that you wouldn't want to boil or pull into a vacuum system.
Bob Holthausen Pall Corporation
} } On Wed, 16 Apr 1997, Ian Montgomery wrote: } } } Like Rob I've been making Reynold's lead citrate since I was a boy. } } I use NaOH pellets but degas the distilled water by sonicating it for a few } } minutes before making the stain. } } } } Ian. } } Leo, If my memory serves I got sonication from the Technical Hints and Tips in the Proceedings of the RMS. I cover my options by sonicating the water whether fresh or hours, days old. Ian.
Does anyone know the Biosupplies address (Melbourne, Australia)? I want to buy 1,3 , 1,4-B-glucan antibody. Thanks in advance. Nuria Cortadellas University of Barcelona
This is an URGENT request for your comments, suggestions, etc about the 3RD RFD for my proposed new immunocytochemistry newsgroup "sci.bio.immunocytochem" now posted in "news.groups" The latter contains material of a rather varied nature (!) but it is necessary to use this unmoderated group to hold the discussions about all the different proposed new groups.
So, if you are connected to Usenet, and you are keen to see a new immunocytochemistry newsgroup, PLEASE go to "news.groups", ignore all the rubbish, look for articles posted on 16.4.97, and you should find my "3RD RFD: sci.bio.immunocytochem" If your not sure how to post articles to Usenet, select "follow-up article" (or equivalent) from your newsreader menu, and post your message so that it appears under the 3RD RFD.
It is VITALALLY IMPORTANT that some discussion takes place in news.groups soon. Although at least 40 people have e-mailed me to say they want the group to happen (THANK-YOU), but only ONE person has posted any response to my RFDs in "news.groups" where the set-up discussion is supposed to take place.
I am concerned that the lack of discussion indicates a lack of interest in the proposed group, and this is why I have postponed holding the vote. But on the 1st May (election day!) I shall be posting my "CALL FOR VOTES" to the people in charge of Usenet newsgroups. A few days after that, you will be able to vote for the proposed new immunocytochem group.
Amanda Wilson Deputy Manager, E.M. Unit St George's Hospital Medical School, S.W.London, UK Tel: 0181 725 5220 (work) e-mail {awilson-at-aw.u-net.com}
This is an URGENT request for your comments, suggestions, etc about the 3RD RFD for my proposed new immunocytochemistry newsgroup "sci.bio.immunocytochem" now posted in "news.groups" The latter contains material of a rather varied nature (!) but it is necessary to use this unmoderated group to hold the discussions about all the different proposed new groups.
So, if you are connected to Usenet, and you are keen to see a new immunocytochemistry newsgroup, PLEASE go to "news.groups", ignore all the rubbish, look for articles posted on 16.4.97, and you should find my "3RD RFD: sci.bio.immunocytochem" If your not sure how to post articles to Usenet, select "follow-up article" (or equivalent) from your newsreader menu, and post your message so that it appears under the 3RD RFD.
It is VITALALLY IMPORTANT that some discussion takes place in news.groups soon. Although at least 40 people have e-mailed me to say they want the group to happen (THANK-YOU), but only ONE person has posted any response to my RFDs in "news.groups" where the set-up discussion is supposed to take place.
I am concerned that the lack of discussion indicates a lack of interest in the proposed group, and this is why I have postponed holding the vote. But on the 1st May (election day!) I shall be posting my "CALL FOR VOTES" to the people in charge of Usenet newsgroups. A few days after that, you will be able to vote for the proposed new immunocytochem group.
Amanda Wilson Deputy Manager, E.M. Unit St George's Hospital Medical School, S.W.London, UK Tel: 0181 725 5220 (work) e-mail {awilson-at-aw.u-net.com}
Please could you publish the following message on your site, in place of the previous "2nd RFD" Thanks!
PROPOSED NEW IMMUNOCYTOCHEMISTRY NEWSGROUP.......UPDATE!
This is an URGENT request for your comments, suggestions, etc about the 3RD RFD for my proposed new immunocytochemistry newsgroup "sci.bio.immunocytochem" now posted in "news.groups" The latter contains material of a rather varied nature (!) but it is necessary to use this unmoderated group to hold the discussions about all the different proposed new groups.
So, if you are connected to Usenet, and you are keen to see a new immunocytochemistry newsgroup, PLEASE go to "news.groups", ignore all the rubbish, look for articles posted on 16.4.97, and you should find my "3RD RFD: sci.bio.immunocytochem" If your not sure how to post articles to Usenet, select "follow-up article" (or equivalent) from your newsreader menu, and post your message so that it appears under the 3RD RFD.
It is VITALALLY IMPORTANT that some discussion takes place in news.groups soon. Although at least 40 people have e-mailed me to say they want the group to happen (THANK-YOU), but only ONE person has posted any response to my RFDs in "news.groups" where the set-up discussion is supposed to take place.
I am concerned that the lack of discussion indicates a lack of interest in the proposed group, and this is why I have postponed holding the vote. But on the 1st May (general election day here in the UK) I shall be posting my "CALL FOR VOTES" to the people in charge of Usenet newsgroups. A few days after that, you will be able to vote for the proposed new immunocytochem group. I need at least 100 YES VOTES for the group to become official. Amanda Wilson Deputy Manager, E.M. Unit St George's Hospital Medical School, S.W.London, UK Tel: 0181 725 5220 (work) e-mail {awilson-at-aw.u-net.com}
The Russian LOMO scopes have a very good (read low) price. Being somewhat leery of Russian quality, has anyone had any experience with them? They have a Physician/Student model for $650.
Mel Dickson wrote: =================================================== I and other collegues have been trying fruitlessly to e-mail Kevex and when I tried to find their web page it was not there either. Is this just a faulty computer or have they been restructured out of existence? Does anyone out there know? =================================================== Try this FAX #: 1-(805)-295-0419.
Kevex is very much alive and well. Two of their top design and manufacturing people stopped by for their annual visit at our exhibit booth at PITTCON 97 a few weeks ago in Atlanta, and they said they were "very busy" and "working hard to keep up". And coming from them, I would expect that was probably 100% correct!
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I have a LOMO low end scope, with added dark field. I am quite pleased with it. It is not as nice to use as a German or Japanese scope, but the optics are quite nice, and you get more of the old fashioned brass, etc. I bought it to use with the kids at home, so I couldn't say how it stands up to industrial use. Built solidly, somewhat homely in paint and finish, but as an optical engineer I am happy with the images.
best regards mark
Mark W. Lund, PhD Director } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"The state is good at simple tasks, like killing people and seizing their wealth. It has far more trouble reaching inside individuals and making them good." Doug Bandow
} I believe that Dr. Mick's interpretation (that REM is german for SEM) is } correct in this case, but there is a technique of Reflection Electron } Microscopy (also abbreviated REM) which is very good for picking out atomic } step edges. See for example: } ... snips
Try also
Reflection Electron Microscopy & Spectroscopy for Surface Analysis Zhong Lin Wang Cambridge University Press ISBN 0 521 48266 6
--------------------------------------------------------------- Dr L. P. Stoter Technical Editor, MICROSCOPY & ANALYSIS Technesis 17, Rocks Park Road email: LPS-at-teknesis.demon.co.uk Uckfield, E. Sussex Phone: +44 (0)1825 766911 TN22 2AT Fax: +44 (0)1825 766911 United Kingdom ---------------------------------------------------------------
I am seeking details of a paper from the 1990 meeting in Seattle. What I have is:
Nicholas G, Bassot J-M, Nicholas M-T (1990). The advantages of cryotechniques: Application to bioluminescent cells. Proceedings of the 12th International Congress of Electron Microscopy, Seattle.
Until we know the details, we can't try for reprint/copy.
This prompts me to ask the obvious question. Does anyone have a foolproof indicator to check the activity of an osmium solution, ideally without processing tissue and viewing in the microscope?
I 'm sure at some time we have all picked up a bottle of expensive osmium and thought do I use it or make fresh up. This happened to me recently and I tried soaking some osmium into a piece of cocktail stick, which appeared to darken, but I was wrong.
Malcolm Haswell University of Sunderland UK ----------
Dare Chris I had the same problem once when I was processing some cultured cells for EM. Morphology was bad at all, but there was no contrast on all membrane. Then I processed the same cell culture with the same batch of osmium solution and fresh osmium side by side. It turned out fresh osmium solved problem.
} I am seeking details of a paper from the 1990 meeting in Seattle. What I } have is: } } Nicholas G, Bassot J-M, Nicholas M-T (1990). The advantages of } cryotechniques: Application to bioluminescent cells. Proceedings of the } 12th International Congress of Electron Microscopy, Seattle. } } Until we know the details, we can't try for reprint/copy
Keith: This was a joint meeting with EMSA and the microbeam society. The proceedings of the meeting published as a four volume set. G.W. Bailey was the publications manager and the San Francisco Press, Inc. published the four volumes. I have Volume 3: Biological Sciences from that meeting. Volume1: Imaging Sciences, Volume 2: Analytical Sciences and Volume 4: Materials Science comprise the titles of the other three volumes. The Nicholas paper was not in volume 3.
The ICEM Program chairs for the meeting were Lee Peachey and D. B. Williams. Good hunting.
Blystone in Texas
-------------------------------- Robert V. Blystone, Ph.D. rblyston-at-trinity.edu
Department of Biology Trinity University 715 Stadium Drive San Antonio, Texas 78212 210.736-7243 FAX 210/736-7229
Dear Colleagues, Does anybody know why everybody uses for staining of ultra thin sections aranyl acetate which has the limited solubility but not uranyl nitrate or uranyl sulfate with the higher solubility.
Sincerely yours, Alexander Mironov
Unit of Morphology Consorzio Mario Negri Sud Via Nazionale, 66030 S. Maria Imbaro (Chieti) Italy
I am a new member on this mailing list. I am receiving replies to the messages that did not submit. And some messages are posted on my mail two or three times. Is this normal?
} I am seeking details of a paper from the 1990 meeting in Seattle. What I } have is: } } Nicholas G, Bassot J-M, Nicholas M-T (1990). The advantages of } cryotechniques: Application to bioluminescent cells. Proceedings of the } 12th International Congress of Electron Microscopy, Seattle. } } Until we know the details, we can't try for reprint/copy
Dear Keith,
I have all five of the volumes from the Seattle meeting. I looked through all of them and I could not find any papers written by Nicholas or Bassot. Are you sure you have the correct meeting?
Good Luck ,
Margaret E. Bisher
NEC Research Institute 4 Independence Way Princeton, NJ 08540. Tel.: (609) 951-2629 Fax: (609) 951-2496 e-mail: peggy-at-research.nj.nec.com
Dear List Is there a reference or does someone have experience with double staining of Epoxy semi-thin sections. I would like to use Basic Fuchsin for red and Toluidine Blue for the blue but I haven't been able to find out any information. Anyone willing to share first hand experience would be appreciated. By the way I am looking for areas with Epstein-Barr viral inclusions in the nucleus. TIA Jack Megill BMS Princeton, NJ
The Nicolas, Bassot, Nicolas talk was the invited lead-off paper in the Cryo-Specimen Prepartation Techniques Symposium chaired buy McDowell and Talon on Thursday August 16, 1990.
The abstract should be in the Imaging Sciences volume unless they did not submit one - unlikely but I recall that this did happen with several invited speakers at the behest of session chairs. This illustrates the problems that inflict the world when guidelines are not enforced.
I will check this volume when I get to where my Proceedings are kept.
There are about 2000 volumes out there so that this matter can be resolved.
} I am seeking details of a paper from the 1990 meeting in Seattle. What I } have is: } } Nicholas G, Bassot J-M, Nicholas M-T (1990). The advantages of } cryotechniques: Application to bioluminescent cells. Proceedings of the } 12th International Congress of Electron Microscopy, Seattle. } } Until we know the details, we can't try for reprint/copy
Dear Keith,
I made a mistake. After seeing Robert Fisher's e-mail I double checked just what I had looked at and I had been looking in the proceedings from the meeting in Paris, Oops!
I did look in the Seattle proceedings and indeed it is there. It is on pages 486 & 487 of Volume 1, Imaging Sciences, just like Dr. Fisher said. If you would like a copy I would be glad to fax it to you or post it in the mail.
Just let me know.
Margaret E. Bisher
NEC Research Institute 4 Independence Way Princeton, NJ 08540. Tel.: (609) 951-2629 Fax: (609) 951-2496 e-mail: peggy-at-research.nj.nec.com
John Megill, Many years ago in graduate school I used to use several different methylene blue-basic fuchsin stains. I remember a special handout provided by LKB that compared these references and showed colored micrographs. They were beautiful. Good Luck!!!! Nancy Monteiro-Riviere
References as follows:
Aparicio, SR, and Marsden, P: Methylene blue-basic fuchsin. Journal of Microsc. (Eng.) 89:139-141, 1969.
Huber, JD, Parker, F, and Odland, GF; Basic fuchsin and alkalinized methylene blue. Stain Technol. 43:83-87, 1968.
Humphrey, CD, and Pittman, Fe: Methylene blue-azure II and basic fuchsin. Stain Technol 42:9-14, 1974.
Nancy A. Monteiro-Riviere, Ph.D.,BCFE,BCFM Professor of Investigative Dermatology/Toxicology North Carolina State University College of Veterinary Medicine Cutaneous Pharmacology and Toxicology Center 4700 Hillsborough Street Raleigh, NC 27606 Telephone: 919-829-4426 FAX: 919-829-4358 email: Nancy_Monteiro-at-ncsu.edu CTPC Homepage: http://cptc.ncsu.edu
We sonicate whatever volume we will need to use in a standard ultrasonic bath. We place solution in a clean beaker and fill the bath with water. Place beaker in the water bath and sonicate for ~15 minutes. If the fluid has a very high viscosity we would sonicate a little longer, maybe 30 minutes.
Bob Holthausen Pall Corporation Scientific and Laboratory Services
williams-at-anatomy.iupui.edu (James C. Williams Jr.) on 04/17/97 03:59:28 PM
To: Microscopy-at-sparc5.microscopy.com -at- internet cc: (bcc: Bob Holthausen/SLSNY/Pall/US)
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} I am seeking details of a paper from the 1990 meeting in Seattle. What I } have is: } } Nicholas G, Bassot J-M, Nicholas M-T (1990). The advantages of } cryotechniques: Application to bioluminescent cells. Proceedings of the } 12th International Congress of Electron Microscopy, Seattle. } } Until we know the details, we can't try for reprint/copy
Dear Keith,
I made a mistake. After seeing Robert Fisher's e-mail I double checked just what I had looked at and I had been looking in the proceedings from the meeting in Paris, Oops!
I did look in the Seattle proceedings and indeed it is there. It is on pages 486 & 487 of Volume 1, Imaging Sciences, just like Dr. Fisher said. If you would like a copy I would be glad to fax it to you or post it in the mail.
Just let me know.
Margaret E. Bisher
NEC Research Institute 4 Independence Way Princeton, NJ 08540. Tel.: (609) 951-2629 Fax: (609) 951-2496 e-mail: peggy-at-research.nj.nec.com
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} I am seeking details of a paper from the 1990 meeting in Seattle. What I } have is: } } Nicholas G, Bassot J-M, Nicholas M-T (1990). The advantages of } cryotechniques: Application to bioluminescent cells. Proceedings of the } 12th International Congress of Electron Microscopy, Seattle. } } Until we know the details, we can't try for reprint/copy
Dear Keith,
I made a mistake. After seeing Robert Fisher's e-mail I double checked just what I had looked at and I had been looking in the proceedings from the meeting in Paris, Oops!
I did look in the Seattle proceedings and indeed it is there. It is on pages 486 & 487 of Volume 1, Imaging Sciences, just like Dr. Fisher said. If you would like a copy I would be glad to fax it to you or post it in the mail.
Just let me know.
Margaret E. Bisher
NEC Research Institute 4 Independence Way Princeton, NJ 08540. Tel.: (609) 951-2629 Fax: (609) 951-2496 e-mail: peggy-at-research.nj.nec.com
This is a protocol we have used that gives wonderful results:
Polychrome stain
1. Blue Stain: Methylene blue .13gm , Azure II .02gm, Glycerol 10ml, Methyl alcohol 10ml, D.H2O 80ml (stir and filter, keeps 6 mo.)
2. Red stain: STOCK SOL: Basic fuchsin .2gm, DH2O 100ml (stir and filter, keeps 6mo.) WORKING SOL: dilute stock 1:4 in DH2O (fresh daily)
3. Sodium Hydroxide 1% fresh daily
Procedure: 1. Flood slide with blue stain 15-60 seconds, depending on temperature and material. 2. Add 4-6 drops NaOH to the stain and mix by tilting the slide, about 10 seconds total time. 3. Wash in running water and dry on hotplate. Blue stain can be destained by heating. 4. Add red stain for 15-30 seconds on hotplate. (Red stain cannot be destained) 5. Rinse with running water and dry.
References: Mackay and Mead. 1970. 28th EMSA meetings pp.296-297. Modified by Griffin and Fahrenbach, Oregon Regional Primate Research Center
Hope this is of some help Linda M. Fox Dept. of CBN and Anatomy Loyola University Medical School 2160 S. First Ave. Maywood, Illinois 60153 lfox1-at-wpo.it.luc.edu
I am a new member on this mailing list. I am receiving replies to the messages that did not submit. And some messages are posted on my mail two or three times. Is this normal?
Hong Yi
Emory, Neurology
Basically, yes. This message has CC: Microscopy-at-Sparc5.Microscopy.Com so it will be sent to the list. It also has To: hyi-at-emory.edu so it will go straight to you. You will probably receive two copies. Typical mailers will put in the CC: header when a "reply" command is issued, so everyone on the list will get replies unless the sender specifically deleted the CC: header before sending the reply.
Sometimes users send in a message, wait an hour to see if it really went in, and then send it again because they haven't seen it yet. This can cause multiple copies of messages. There are other possible causes as well. I noticed a few duplicates recently but didn't spend the time to figure out how they happened.
I was wondering if anyone knew of a good fix for TEM studies of ciliated protists in marine organisms such as oysters. I've tried different fixes and the tissue looks good but the protists....uhhhh looks bad. Any suggestions? Thanks.
Peace through Christ,
Phil Rutledge, Director e-mail: prutle1-at-gl.umbc.edu Center for Microscopy voice: (410) 455-3582 UMBC Dept. of Biology fax: (410) 455-3875 Catonsville, MD 21228 ///// / / / / /////// /////// / / /////// /////// / / / / / / / / / / / / /////
Received: by dub-img-5.compuserve.com (8.6.10/5.950515) id PAA07822; Fri, 18 Apr 1997 15:07:01 -0400 Comments: Returned from: {74767.3673-at-CompuServe.COM} Message-Type: Delivery Report Message-ID: {970418190028_515664.456256_GHQ43-46-at-CompuServe.COM}
This is a protocol we have used that gives wonderful results:
Polychrome stain
1. Blue Stain: Methylene blue .13gm , Azure II .02gm, Glycerol 10ml, Methyl alcohol 10ml, D.H2O 80ml (stir and filter, keeps 6 mo.)
2. Red stain: STOCK SOL: Basic fuchsin .2gm, DH2O 100ml (stir and filter, keeps 6mo.) WORKING SOL: dilute stock 1:4 in DH2O (fresh daily)
3. Sodium Hydroxide 1% fresh daily
Procedure: 1. Flood slide with blue stain 15-60 seconds, depending on temperature and material. 2. Add 4-6 drops NaOH to the stain and mix by tilting the slide, about 10 seconds total time. 3. Wash in running water and dry on hotplate. Blue stain can be destained by heating. 4. Add red stain for 15-30 seconds on hotplate. (Red stain cannot be destained) 5. Rinse with running water and dry.
References: Mackay and Mead. 1970. 28th EMSA meetings pp.296-297. Modified by Griffin and Fahrenbach, Oregon Regional Primate Research Center
Hope this is of some help Linda M. Fox Dept. of CBN and Anatomy Loyola University Medical School 2160 S. First Ave. Maywood, Illinois 60153 lfox1-at-wpo.it.luc.edu
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Re:
} Nicholas G, Bassot J-M, Nicholas M-T (1990). The advantages of } cryotechniques: Application to bioluminescent cells. Proceedings of the } 12th International Congress of Electron Microscopy, Seattle.
Dear Robert,
Why not write directly to Marie-Therese Nicholas at Laboratoire de Bioluminescence CNRS 105 Blvd Raspail 75006 Paris France
As an author, she should be able to help you.
Regards,
Paul Webster, Ph.D Center for Cell Imaging Yale School fo Medicine http://info.med.yale.edu/cellimg
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Re:
} Nicholas G, Bassot J-M, Nicholas M-T (1990). The advantages of } cryotechniques: Application to bioluminescent cells. Proceedings of the } 12th International Congress of Electron Microscopy, Seattle.
Dear Robert,
Why not write directly to Marie-Therese Nicholas at Laboratoire de Bioluminescence CNRS 105 Blvd Raspail 75006 Paris France
As an author, she should be able to help you.
Regards,
Paul Webster, Ph.D Center for Cell Imaging Yale School fo Medicine http://info.med.yale.edu/cellimg
I've had no problems with Reynold's lead citrate, using freshly boiled, distilled water and 1N NaOH freshly prepared from pellets. I store the finished stain in a disposable syringe with all air bubbles expelled, and the tip covered in plastifilm. For use I attach a 0.2 micron syringe filter and needle and expel a few drops first, then use the next drops. As long as there is no air in the syringe, and it is kept in the refrig, it will keep forever? or at least 8 months. The recipe: 1.33 g lead nitrate, 1.76 g sodium citrate mixed with 30 ml freshly boiled distilled or deionized water- mix in a 50 ml volumetric flask- shake vigorously for 1 min and intermittently for 30 mins. Solution will be milky.Add 8.0 ml of 1 N NaOH freshly prepared.Solution becomes clear. Dilute to 50 ml with freshly boiled, distilled or deionized water. Load into syringe- I use a 60 cc disposable distilled or deionized water. Load into syringe- I use a 60 cc disposableI,ve been using this method for 20 years consistent success. Just my two-cents worth.
Pat Masarachia Bone Biology Merck and Co. West Point, PA 215-652-7999
So why do mail problems happen on Friday Afternoons?
Maybe that 's when accounts are deleted or something. I've been doing some investigation on this afternoons set of problems. There was an intermittent fault in mail duplicaton. If your interested read on, otherwise delete this and just consider this as just a test message.
The duplicate transmission was caused by the mail nameserver hanging on a memory/queue failure. This was due to a number of sites (~ 10) either going down and/or just going out of existance (host name unkown) plus a number of user accounts either filling (mailbox full) or being deleted (user unknown) without the user unsubscribing (~20) . The net effect of all these was that the queue of undelivered messages started to grow significantly. This in turn started a new set of delivery queuese which began to eat up even more CPU resources.
Eventually a few of the queues failed and but when message queue fails the sendmail program tries to resend the entire message, however, it (sendmail program) forgot that it had sent the message to part of the mailing list already and so a large number of people got multiple copies, depending upon where you name feel in the subscription list.
I've tried to reconfigure the server to minimize this happening again, however, to do so I've instituted yet another queuing system for all listserver mail. Instead of trying to immediately deliver all mail as we did in the past, everything is now put into a small queue and delayed ~ 15-30 minutes before starting a delivery sequence.
This does not mean that you will see a message 15-30 minutes after someone submits it, but the process of running the mail server becomes more regimented.
Due to the size of the mailing list and the sometime poor links to some sites mail can take more than an hour (or longer) to propagate through the entire list. I've also attempted to setup a mail configuration file to minimize duplicate deliveries in the case of a queue failure. If things work correctly no more than 10 people should receive duplicate messages on a crash, as the delivery list is rewritten now every 10 deliveries. This will also (obviously) increase the time it takes to process the mail.
Let's see if this cures the duplicate delivery problem... (fingers crossed and blurry eye he reaches for the send button)
G'night all.
Nestor Yawn.. Your tired Friendly Neighborhood SysOp
Q: How many internet mail list subscribers does it take to change a light bulb?
A: 1,331:
1 to change the light bulb and to post to the mail list that the light bulb has been changed. 14 to share similar experiences of changing light bulbs and how the light bulb could have been changed differently. 7 to caution about the dangers of changing light bulbs. 27 to point out spelling/grammar errors in posts about changing light bulbs. 53 to flame the spell checkers. 156 to write to the list administrator complaining about the light bulb discussion and its inappropriateness to this mail list. 41 to correct spelling in the spelling/grammar flames. 109 to post that this list is not about light bulbs and to please take this email exchange to alt.lite.bulb. 203 to demand that cross posting to alt.grammar, alt.spelling and alt.punctuation about changing light bulbs be stopped. 111 to defend the posting to this list saying that we are all use light bulbs and therefore the posts **are** relevant to this mail list. 306 to debate which method of changing light. bulbs is superior, where to buy the best light bulbs, what brand of light bulbs work best for this technique, and what brands are faulty. 27 to post URLs where one can see examples of different light bulbs. 14 to post that the URLs were posted incorrectly, and to post corrected URLs. 3 to post about links they found from the URLs that are relevant to this list which makes light bulbs relevant to this list. 33 to concatenate all posts to date, then quote them including all headers and footers, and then add "Me Too.". 12 to post to the list that they are unsubscribing because they cannot handle the light bulb controversy. 19 to quote the "Me Too's" to say, "Me Three.". 4 to suggest that posters request the light bulb FAQ. 1 to propose new alt.change.lite.bulb newsgroup. 47 to say this is just what alt.physics.cold_fusion was meant for, leave it there.
I've received enough requests for copies of the general information and FAQ messages that I've now put them up on-line for you to access via the WWW at your convenience.
Just go to the MSA WWW home page and follow the Microscopy ListServer Links.
http://www.msa.microscopy.com
I will continue to send out Email versions with every new subscription or to anyone requesting an Emailed copy.
Nestor Your Friendly Neighborhood SysOp.
-------- BTW,... (with fingers crossed) no duplicate messages so far!
I am study artifacts on trasmission electron microscopy. I have not bibliography. Please your send references. Thank you Dr. Alberto Pizarro G. Histopathology rediegal-at-homonet.com.mx
Dear Colleagues, Does anybody know why for the staining of ultra thin sections everybody uses uranyl acetate which has rather limited solubility in water, but not yranyl sulfate or uranyl nitrate which are much more soluble.
Alexander Mironov
Unit of Morphology Consorzio Mario Negri Sud S. Maria Imbaro (Chieti) Italy
} This prompts me to ask the obvious question. Does anyone have a foolproof } indicator to check the activity of an osmium solution, ideally without } processing tissue and viewing in the microscope? } } I 'm sure at some time we have all picked up a bottle of expensive osmium } and thought do I use it or make fresh up. This happened to me recently and I } tried soaking some osmium into a piece of cocktail stick, which appeared to } darken, but I was wrong. } } Malcolm Haswell
Malcolm, I would put a drop of corn oil (maize oil on your side of the puddle) on a slide and some of the osmium solution in a small dish (size so that the slide acts as a lid for the dish). If the osmium is good, vapors from it will blacken the oil droplet. (Any polyunsaturated oil will do.) Phil
&&& Illigitimi non carborundum &&&&&&&& Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217) 355-1143 oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
Seattle paper: thanks to everyone for the help in finding details of the paper in question. It was found and offers made of a copy!
Flat beer: most of this was off line - apologies for that which crept back on-line. The result is I am bearing home brew kits to a new found friend in Chicago next month! So, some good came of it!
Thanks again from Plymouth UK (on a grey Monday morn) Keith Ryan
10 BIT MONOCHROME } 62dB S/N REAL TIME CAMERAS, FRAME GRABBERS, & SOFTWARE INFO / NEW WEB SITE DVC Company is a US manufacturer of video cameras and has a updated web with detailed info /Q & A on: DVC-10 10 bit digital & 10 bit analog monochrome CCD cameras DVC-8 8 bit digital & 10 bit analog monochrome CCD cameras DVC-0A 10 bit analog monochrome camera (upgradable) to digital Complete frame grabber PCI bus board listing supplied by DVC as a systems house, along with imaging software. ((( DVC can offer a complete package ))) camera, digital cable, frame grabber, software, and tuneable monochrome and RGB LCD filters http://members.aol.com/dvcco or http://www.edt.com/dvc/dvc.html Fill in the form for contact and or: Download the complete DVC 80 page manual, data sheet, and TI sensor info via Acrobat 3.0 reader for selective print out later. DVC Company/ San Diego, CA 619-444-8300 619-444-8321-fax This posting is only on two newsgroups due to the focus of the products.
I am mailing you regarding membership to the American Society of Electron Microscopy. Arthur Slaughter and myself are electron microscopists at HRL Technology, Melbourne Australia, and are currently members of the Australian Society for Electron Microscopy. HRL Technology is a Research and Development laboratory which was originally part of the State Electricity Commission of Victoria. The Electron Microscope Facility at HRL is mainly used for solving materials related problems for a wide range of industries in Australia, including mineral and metals processing, mining and power production, building and food industries to name a few.
We are interested in subscribing to the American Society of Electron Microscopy as we believe this would benefit our day to day work, and provide information regarding current areas of concern in the US.
Could you please send us information regarding ASEM, and any conditions required for membership to this organisation.
MAMAS (Mid-Atlantic Microbeam Analysis Society) and the Surface and Microanalysis Science Division, NIST Meetin at the National Institute of Standards and Technology, Gaithersburg, MD on Thursday, May 15, 1997, 10:30 am- 3:00 PM Lecture Room D, Administration Bldg.
10:30am Coffee and Doughnuts
10:45am Prof. David R. Veblen, Dept. of Earth and Planetary Science, Johns Hopkins University "Transmission Electron Microscopy of Minerals"
12 noon Lunch
1:15pm Dr. Michael Kersker, TEM/STM Project Manager, JEOL USA "200 KV FEG: Refried Beans with a Fiery New Salsa"
For more information, contact Ryna Marinenko (301)975-3901, FAX(301)417-1321, email:ryna.marinenko-at-nist.gov
We work as consultants for a computer imaging hardware and software manufacturer's rep, and have had a rather unusual request. Perhaps someone, either end users or vendors can assist.
We are looking for the equivalent of a MacBeth Color Chart, typically used in video, but this needs to be very small, translucent, and mounted on a standard histo glass slide. Ideally, 1/32" square (1 mm x 1 mm OK) and it would contain at least the primary colors, incl. black and white. I suspect there may be a problem with ?translucent black and white. I think episcopic illumination would also suffice for the chart, however so it could be opaque. At this time, I do not have more detail on their exact application.
We have checked with Munsell and MacBeth to no avail, so we would appreciate any assistance, and even a referral as it's likely this is a custom application. You can e-mail me directly if you like.
I am sending this message again because it was returned, in error on Friday. ----------
This prompts me to ask the obvious question. Does anyone have a foolproof indicator to check the activity of an osmium solution, ideally without processing tissue and viewing in the microscope?
I 'm sure at some time we have all picked up a bottle of expensive osmium and thought do I use it or make fresh up. This happened to me recently and I tried soaking some osmium into a piece of cocktail stick, which appeared to darken, but I was wrong.
Malcolm Haswell University of Sunderland UK ----------
Dare Chris I had the same problem once when I was processing some cultured cells for EM. Morphology was bad at all, but there was no contrast on all membrane. Then I processed the same cell culture with the same batch of osmium solution and fresh osmium side by side. It turned out fresh osmium solved problem.
Message-ID: {01BC4E50.DFE665E0-at-pguerin.lisco.com} {CONFOCAL-at-LISTSERV.ACSU.BUFFALO.EDU} , "'MICROSCOPY-at-SPARC5.MICROSCOPY.COM'" {MICROSCOPY-at-Sparc5.Microscopy.Com} Cc: Chris MacLean {Windows/pguerin/cmaclean-at-vaytek.com}
This message is from a software vendor.
Hello,
We're in the process of defining specifications for the next version of = VoxBlast. For those of you who are not familiar with VoxBlast, it is a = 3D reconstruction, volume visualization and measurment software running = on UNIX, Windows, and Mac.
Since this product is for you, it would be to our mutual benefit if you = let us know what you'd like to see in future versions of VoxBlast. The = most useful feedback, describes the improvement in detail.
Thank you in advance for your time.
Best regards
Patrick Guerin Customer Technical Support Engineer VayTek, Inc. 305 West Lowe Suite 109 PO Box 732 Fairfiled Iowa 52556-0732
Group - I would like to organize a summary listing of all non-supplier/manufacturer web pages of interest to microscopists. I already do the supplier/manufacturer listing. Format to be WWW address, organizers name and establishment and a some-100 max word description of the site. When complete I will publish the full listing on this listserver as well as in my publication. All help would be much appreciated. Don Grimes, Microscopy Today
Maybe y'all can help this soul out until he can get subscribed.
} Return-Path: {muellerd-at-mis.finchcms.edu} } Date: Mon, 21 Apr 1997 14:55:43 -0700 } From: David Mueller {muellerd-at-mis.finchcms.edu} } Reply-To: muellerd-at-mis.finchcms.edu } Organization: The Chicago Medical School } To: sdw-at-biotech.ufl.edu } Subject: collodial gold labeling } } I am uncertain how to access this newsgroup on EM. Could you give me } the } address? } } Alternatively, maybe you know the answer. I need to label MAb with } colloidal gold. Since the Ab is available in only small amount, I } prefer not to optimize the conditions as it requires a lot of Ab. Are } there standard conditions to label MAb with gold? What is the minimal } protein concentration needed to get effective binding? Can BSA be added } to stabilize the binding and fill the unbound sites? } } Thanks for any help in the matter. } } David Mueller } Dept. Biological Chemistry } The Chicago Medical School } muellerd-at-mis.finchcms.edu } }
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 352-392-1295 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
A colleage of mine, Lonnie Russell, has co-authored an introductory book on molecular biology (MB) that may be of interest to microscopists. Since MB may be new or intimidating (yet is important for microscopists) this might be one book to consider. If you would like more information, he may be contacted directly at {lrussell-at-som.siu.edu} . Note: I have no financial interest in this book - I just find it useful.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
I am looking into upgrading our present image Analysis system (Kontron IBAS).
Has anyone performed extensive comparisons between the leading IA manufacturers who is willing offer pro's and con's of each IA system they have examined.
Some of the IA systems include (but not limited to)
Kontron KS400 Optimas Mediacybernetics Image Pro Plus ImagePro (Nikon) Bioquant Quantimat Noesis (Visilog5)
Applications include wide variety of biological (Pathology/toxicology applications, density gradients, fluorescence, stereology and morphometry etc.), and non biological (particle size, coating thickness, fiber analysis, phase boundary etc.).
Ease of programming (Macro scripting or interpreter language) and program modification for those who are not computer programers is a must.
Input on camera and video capture boards are welcomed as well.
Thank you in advance for any information you are willing to share.
The heaviest element known to science was recently discovered. The element, tentatively named ADMINISTRATIUM, has no protons or electrons and thus has an atomic number of 0.
However, it does have 1 Neutron, 128 Assistant Neutrons, 75 Vice-Neutrons and 111 Assistant Vice Neutrons. This gives it an atomic weight of 315. These 315 particles are held together in a nucleus by a force that involves the continuous exchange of meson-like particles called Morons.
Since it has no electrons, Administratium is inert. However, it can be detected chemically as it impedes every other reaction with which it comes into contact. According to the discoverers, a minute amount of Administratium caused one reaction to take over four days to complete, when it would normally occur in less than one second.
Administratium has a normal life of approximately 3 years, at which time it does not decay, but instead, undergoes a reorganization in which Assistant Neutrons, Vice-Neutrons and Assistant Vice-Neutrons exchange places. Some studies have shown that the atomic weight actually increases after each reorganization.
Research at other laboratories indicates that Administratium occurs naturally in the atmosphere. It tends to concentrate at certain points such as government, large companies, healthcare facilities and universities; and will often be found in the newest, best maintained buildings.
Scientists point out that Administratium is know to be toxic at any level of concentration and can easily destroy any productive reactions where it is allowed to accumulate.
Sad but all too true !!
Sent to me by
JamesR0712-at-aol.com \\|// (o o) ~~~~~~~~~~~~oOOo~(_)~oOOo~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
{ { {This message is made of 100% recycled electrons} } }
Cheers ;o) :o) %o) Eric {Mesa Arizona} http://www.goodnet.com/~earosen (Note the tilde before earosen)
Do you really need to label your antibody? Rather, I'd recommend that you use an indirect labelling method in which, e.g., the secondary antibody is coupled to gold or is biotinylated and finally detected with streptavidin gold.
If you really need to gold label your MAb, you will find the necessary information and protocols in the reviews and papers published by J. Roth in the late 70's and early 80's, as well as in many books on immunolabelling.
Regards, Michel
**************************************************** Michel Deschuyteneer, Ph.D. deschuyt-at-sbbio.be Scientist Electron Microscopy Laboratory
SmithKline Beecham Biologicals Rue de l'Institut, 89 B1330 Rixensart, BELGIUM Tel: +32-2-656 9290 Fax: +32-2-656 8164 **************************************************** Standard disclaimer: the opinions expressed in this communication are my own and do not necessarily reflect those of SmithKline Beecham. ****************************************************
I am looking for the fax number or email address of Scanning Microscopy Int. (Chicago) - can anybody help?
Thanks in advance Fiona Graham Electron Microscope Unit University of Natal, Dalbridge, 4041, South Africa tel: +27 31 260 2174 fax: +27 31 261 6550 email: GRAHAM-at-ph.und.ac.za URL: http://www.und.ac.za/und/emu/emunit.html
You wrote: " I am looking for the fax number or email address of Scanning Microscopy Int. (Chicago) - can anybody help?
Thanks in advance Fiona Graham Electron Microscope Unit University of Natal, Dalbridge, 4041, South Africa tel: +27 31 260 2174 fax: +27 31 261 6550 email: GRAHAM-at-ph.und.ac.za"
I have used 73211.647-at-compuserve.com within the last couple of months and it was OK.
Donald J. Marshall Relion Industries P.O. Box 12 Bedford, MA 01730 Ph: 617-275-4695 FAX: 617-275-4695 Alternate FAX: 617-271-0252
DMartin/RRosencrans wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Hello all, } } We work as consultants for a computer imaging hardware and software } manufacturer's rep, and have had a rather unusual request. Perhaps someone, } either end users or vendors can assist. } } We are looking for the equivalent of a MacBeth Color Chart, typically used } in video, but this needs to be very small, translucent, and mounted on a } standard histo glass slide. Ideally, 1/32" square (1 mm x 1 mm OK) and it } would contain at least the primary colors, incl. black and white. I suspect } there may be a problem with ?translucent black and white. I think episcopic } illumination would also suffice for the chart, however so it could be } opaque. At this time, I do not have more detail on their exact application. } } We have checked with Munsell and MacBeth to no avail, so we would appreciate } any assistance, and even a referral as it's likely this is a custom } application. You can e-mail me directly if you like. } } Thanks in advance!! } } Daryl Martin } dmartin-at-ic.net } } (313) 213-8444Dear Daryl, When I worked at Zeiss as microspectrophotometry specialist, one of my colleagues made a wonderful test slide using very small strips of colored film, laid side by side on a slide then covered with a coverslip. It sounds like a modification would be ideal for your purposes.
Hope this helps. Barbara Foster Consortium President Microscopy/Marketing & Education
Gregory.Argentieri-at-sandoz.com wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Dear Microscopists: } } I am looking into upgrading our present image Analysis system (Kontron } IBAS). } } Has anyone performed extensive comparisons between the leading IA } manufacturers who is willing offer pro's and con's of each IA system } they have examined. } } Some of the IA systems include (but not limited to) } } Kontron KS400 } Optimas } Mediacybernetics Image Pro Plus } ImagePro (Nikon) } Bioquant } Quantimat } Noesis (Visilog5) } } Applications include wide variety of biological (Pathology/toxicology } applications, density gradients, fluorescence, stereology and } morphometry etc.), and non biological (particle size, coating } thickness, fiber analysis, phase boundary etc.). } } Ease of programming (Macro scripting or interpreter language) and } program modification for those who are not computer programers is a } must. } } Input on camera and video capture boards are welcomed as well. } } Thank you in advance for any information you are willing to share. } } Greg } } Gregory Argentieri } Novartis Pharmaceuticals Corp. } Gregory.Argentieri-at-pharma.novartis.com } Gregory.Argentieri-at-sandoz.com } Greg2NJ-at-aol.com } } 201-503-8617Dear Greg, The list you sent suggests that you may want to clarify your needs a little further before you go shopping. 1. Do you want a fully integrated system or something which is more modular? Quantimat and Kontron systems are sold as systems whereas Media Cy, Optimas, and Noeisis are software only. 2. You mentioned the level of programming. I am more familiar with the off-the-shelf systems and of those, Noesis is the most powerful but requires the greatest sophistication in terms of programming ability. Media Cy's Image Pro Plus and Optimas are more mid-range products. We have conducted a variety of market research projects, both at meetings (Microscopy & MicroAnalysis, ASMaterials, Experimental Bio, etc.), by phone, and personal interview. Of these two packages, Media Cy's Image Pro is the more widely accepted software. Several system integrators also indicated that it is more user friendly and the package has been adopted by a number of the microscope companies (ex: Zeiss' Image One is a privately labeled version; I expect that Nikon's offering is the same, Topcon and Phillips have used IPP in conjunction with their EMs). 3. Re: the hardware - we strongly suggest that you talk to a few local system integrators. They can give you the latest on the hardware/software/camera technologies and put together a system which meets your needs. (Input from other colleagues never hurts, however).
Hope this helps. If you need training when you get your system together, give us a call.
Barbara Foster Consortium President Microscopy/Microscopy Education
We have no financial interest in any of the products mentioned above.
Here is a request I am forwarding for a colleague, Daniel Wilcox. Thank you for your help.
} Audrey Dow } } I am looking for experienced Hitachi 4500-II FESEM users that have used } their instruments for electron beam induced current (EBIC) imaging of } gallium arsendie microwave devices. I am looking for advice on how to } build or buy a sub-stage for the large Hitachi stage (with a 6" } intro-port) that will allow for mechanical probe tips to contact device } circuit elements, and allow DC bias to be applied as well as the induced } current signal to be fed to our GW Electronics speciman current amp. I } have done extensive EBIC with our Cambridge 250 SEM, but the amount of } sample current available in the Hitachi is much less. } } If anyone has some suggestions, please contact me at 301-428-4233 at } M/A-COM, e-mail "wilcoxd-at-macom.com". Thanks! } } Daniel Wilcox
(Since Daniel sometimes has trouble with e-mail, you can send messages to me at audrey.dow-at-amp.com and I will forward them.) }
The printing inks used to create the MacBeth Color Chart cannot be precisely duplicated with transparency materials. It is possible to create a close approximation by carefully photographing a color chart with 35mm transparency film. I have used a Kodachrome slide of the MacBeth chart included with a Beseler slide duplicator for calibration of that system. If you are photographing your own chart the most rigorous results will be obtained if you check the slides with a densitometer and correct for any density shifts or color shifts then rephotograph. Once you have a good 35mm slide then you can reduce the size by carefully rephotographing using the same type of controls as above. Hopefully you can work with a good professional film processing laboratory. Their knowledge of color materials and sensitometry can be quite helpful. Keep in mind that film materials vary in their permanency. Some will fade non-linearly quite rapidly when exposed to intense light levels. Larry D. Ackerman (415) 476-8751 Howard Hughes Medical Institute FAX (415) 476-5774 UCSF, Box 0724, Rm U426 533 Parnassus Ave. mishot-at-itsa.ucsf.edu San Francisco, CA 94143
The Conservation Analytical Laboratory (CAL) of the Smithsonian Institution, located in Suitland, MD, is recruiting for a Microscopist, GS-11 ($38,330-$49,831) or GS-12 ($45,919-59,725). Duties include optical microscopy, chemical microscopy, freezing-heating stage analysis, digital imaging and analysis, sample preparation, and microscope maintenance. The incumbent will be required to actively participate in the educational/training activities of CAL.
At least an undergraduate degree in physical/biological science or related field is required. The applicant must have knowledge of the principles and experience in the practice of optical microscopy, and skill in conducting training in a technical field to a professional audience.
For a copy of the vacancy announcement, call the Smithsonian 24-hour Automated Jobline (202) 287-3102, press 9 and request Announcement # 97PL-3079. Applications must be postmarked by June 18, 1997. The Smithsonian Institution is an Equal Opportunity Employer.
} ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Gregory.Argentieri-at-sandoz.com wrote: } } } } } ------------------------------------------------------------------------} } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } } -----------------------------------------------------------------------.} } } } Dear Microscopists: } } } } I am looking into upgrading our present image Analysis system } (Kontron } } IBAS). } } } } Has anyone performed extensive comparisons between the } leading IA } } manufacturers who is willing offer pro's and con's of each IA } system } } they have examined. } } } } Some of the IA systems include (but not limited to) } } } } Kontron KS400 } } Optimas } } Mediacybernetics Image Pro Plus } } ImagePro (Nikon) } } Bioquant } } Quantimat } } Noesis (Visilog5) } } } } Applications include wide variety of biological } (Pathology/toxicology } } applications, density gradients, fluorescence, stereology and } } } morphometry etc.), and non biological (particle size, coating } } } thickness, fiber analysis, phase boundary etc.). } } } } Ease of programming (Macro scripting or interpreter language) } and } } program modification for those who are not computer } programers is a } } must. } } } } Input on camera and video capture boards are welcomed as } well. } } } } Thank you in advance for any information you are willing to } share. } } } } Greg } } } } Gregory Argentieri } } Novartis Pharmaceuticals Corp. } } Gregory.Argentieri-at-pharma.novartis.com } } Gregory.Argentieri-at-sandoz.com } } Greg2NJ-at-aol.com } } } } 201-503-8617Dear Greg, } The list you sent suggests that you may want to clarify your needs a } } little further before you go shopping. } 1. Do you want a fully integrated system or something which is more } modular? Quantimat and Kontron systems are sold as systems whereas } Media } Cy, Optimas, and Noeisis are software only. } 2. You mentioned the level of programming. I am more familiar with } the } off-the-shelf systems and of those, Noesis is the most powerful but } requires the greatest sophistication in terms of programming } ability. } Media Cy's Image Pro Plus and Optimas are more mid-range products. } We } have conducted a variety of market research projects, both at } meetings } (Microscopy & MicroAnalysis, ASMaterials, Experimental Bio, etc.), } by } phone, and personal interview. Of these two packages, Media Cy's } Image } Pro is the more widely accepted software. Several system } integrators } also indicated that it is more user friendly and the package has } been } adopted by a number of the microscope companies (ex: Zeiss' Image } One is } a privately labeled version; I expect that Nikon's offering is the } same, } Topcon and Phillips have used IPP in conjunction with their EMs). } 3. Re: the hardware - we strongly suggest that you talk to a few } local } system integrators. They can give you the latest on the } hardware/software/camera technologies and put together a system } which } meets your needs. (Input from other colleagues never hurts, } however). } } Hope this helps. If you need training when you get your system } together, } give us a call. } } Barbara Foster } Consortium President } Microscopy/Microscopy Education } } We have no financial interest in any of the products mentioned } above.
Greg - since you are upgrading you might also look at Amerinex's Aphelon software. It seems to be very powerful being a spinoff of some AI software developed for the military. I believe that they are at http://www.amerinex.com. If you are willing to program a system youmight also look at ContextVision's MicroGOP - they are from Sweden and the only number I have is there - 46-13-102480 or FAX 46-13-104282.
{BLOCKQUOTE TYPE=CITE} ------------------------------------------------------------------------ {BR} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Gregory.Argentieri-at-sandoz.com wrote: } } } } } ------------------------------------------------------------------------} } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } } -----------------------------------------------------------------------.} } } } Dear Microscopists: } } } } I am looking into upgrading our present image Analysis system } (Kontron } } IBAS). } } } } Has anyone performed extensive comparisons between the } leading IA } } manufacturers who is willing offer pro's and con's of each IA } system } } they have examined. } } } } Some of the IA systems include (but not limited to) } } } } Kontron KS400 } } Optimas } } Mediacybernetics Image Pro Plus } } ImagePro (Nikon) } } Bioquant } } Quantimat } } Noesis (Visilog5) } } } } Applications include wide variety of biological } (Pathology/toxicology } } applications, density gradients, fluorescence, stereology and } } } morphometry etc.), and non biological (particle size, coating } } } thickness, fiber analysis, phase boundary etc.). } } } } Ease of programming (Macro scripting or interpreter language) } and } } program modification for those who are not computer } programers is a } } must. } } } } Input on camera and video capture boards are welcomed as } well. } } } } Thank you in advance for any information you are willing to } share. } } } } Greg } } } } Gregory Argentieri } } Novartis Pharmaceuticals Corp. } } Gregory.Argentieri-at-pharma.novartis.com } } Gregory.Argentieri-at-sandoz.com } } Greg2NJ-at-aol.com } } } } 201-503-8617Dear Greg, } The list you sent suggests that you may want to clarify your needs a } } little further before you go shopping. } 1. Do you want a fully integrated system or something which is more } modular? Quantimat and Kontron systems are sold as systems whereas } Media } Cy, Optimas, and Noeisis are software only. } 2. You mentioned the level of programming. I am more familiar with } the } off-the-shelf systems and of those, Noesis is the most powerful but } requires the greatest sophistication in terms of programming } ability. } Media Cy's Image Pro Plus and Optimas are more mid-range products. } We } have conducted a variety of market research projects, both at } meetings } (Microscopy & MicroAnalysis, ASMaterials, Experimental Bio, etc.), } by } phone, and personal interview. Of these two packages, Media Cy's } Image } Pro is the more widely accepted software. Several system } integrators } also indicated that it is more user friendly and the package has } been } adopted by a number of the microscope companies (ex: Zeiss' Image } One is } a privately labeled version; I expect that Nikon's offering is the } same, } Topcon and Phillips have used IPP in conjunction with their EMs). } 3. Re: the hardware - we strongly suggest that you talk to a few } local } system integrators. They can give you the latest on the } hardware/software/camera technologies and put together a system } which } meets your needs. (Input from other colleagues never hurts, } however). } } Hope this helps. If you need training when you get your system } together, } give us a call. } } Barbara Foster } Consortium President } Microscopy/Microscopy Education } } We have no financial interest in any of the products mentioned } above.
Greg - since you are upgrading you might also look at Amerinex's Aphelon software. It seems to be very powerful being a spinoff of some AI software developed for the military. I believe that they are at http://www.amerinex.com. If you are willing to program a system youmight also look at ContextVision's MicroGOP - they are from Sweden and the only number I have is there - 46-13-102480 or FAX 46-13-104282.
{BLOCKQUOTE TYPE=CITE} ------------------------------------------------------------------------ {BR} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
4/22/97 4:25 PM Ultramicroscopy
Does anyone know what's become of the Elsevier journal "Ultramicroscopy?" They have apparently not published a single issue in 1997, although they normally appear monthly. An inquiry with the publisher was answered with a brief statement "Thank you for your inquiry. Unfortunately, the journal has not yet been published for 1997." I would withdraw my paper so that it could be published elsewhere (it was accepted in December 1996, and I even reviewed the galley proofs)... but it was supposed to be part of a conference proceedings issue (the Sixth Frontiers Conference, June 1996), so I would like to find out more before doing anything.
Still working in the dark... Jeff Fortner Argonne National Laboratory Chemical Technology Division Argonne, IL 60440
I am looking for a film scanner (digitizer for HRTEM films), which has resolution better than 2000 pixels/inch and 12 or more bits per pixel. I found one from Kodak's products, but it is too expensive (it works for color film, too). Anyone who has suggestion please email to panx-at-umich.edu Thank you.
Xiaoqing
_______________ Xiaoqing Pan, Ph.D. Associate Professor, Materials Science & Engineering University of Michigan 2038 H. H. Dow Building 2300 Hayward Street Ann Arbor, MI 48109-2136
Xiaoqing Pan wrote: } } I am looking for a film scanner (digitizer for HRTEM films), which } has resolution better than 2000 pixels/inch and 12 or more bits } per pixel. I found one from Kodak's products, but it is too expensive.
Dear Xiaoqing,
I have used an Agfa SelectScan, with specs (from my local distributor): Optical Resolution 4000 ppi x 4000 ppi Optical Density Range 3.6 Bits per pixel 13 (samples at 16) Cost ~A$72,000
I had no trouble with the digitised images from this scanner (other than the amount of RAM required to manipulate them!) ... a satisfied customer.
Given the high price of the SelectScan, another option might be the Agfa Vision 35: Optical Resolution 3175 ppi x 3175 ppi Optical Density Range 3.2 Bits per pixel 12 Cost ~A$12,000
I have no commercial interest in either of these products.
Stephen. ............................................................... : Stephen Anderson Australian Key Centre for : : Microscopy and Microanalysis : : Email stephen-at-emu.usyd.edu.au The University of Sydney : : Telephone (+61)-2-9351 7552 NSW 2006 : : Facsimile (+61)-2-9351 7682 Australia : :.............................................................:
We will hopefully get funding soon for a mid range voltage (300, 400 kV) TEM such as the H-9000, 3010, 4010 or CM300. We are very interested in obtaining private warts and all reports from operators of such microscopes on their satisfaction with their purchase.
We are interested in answers to questions such as:
What microscope did you choose and what was the main reason for your selection?
From the start of installation, how many days were taken before resolution was confirmed?
In days per year, how often is the microscope unusable with instrument failure?
Have there been any catastrophic ($10,000 +) failures?
Is it easy to maintain specified resolution?
How easy is it to train users to operate?
Would your microscope fit easily into a multi-user laboratory?
Are there any serious design problems you know of?
Would you buy another microscope from the same maker?
How would you rate your overall satisfaction with your microscope?
1 2 3 4 5 very unsatisfied unsatisfied just satisfied quite satisfied ecstatic
Mail replies to me, rather than posting them. I will collate replies and if anyone else wants to know the score, they can mail me for a private copy of the report.
Mel Dickson Director, E.M. Unit, University of New South Wales, Sydney Australia
At HRL we are urgently seeking a circuit board for the Motorised Stage Drive (ISM-MSD40-2) for our Jeol 840 SEM. The required circuit board is the computer interface (RS232) for external control, ie a EDXA control of the SEM stage. Should anyone know the whereabouts of a spare board or can get the schematic of this board we will be glad to here from you.
I have a colleague who would like to know if there is a way to deside if bacteria in activated sludge are gram-positive or gram-negative. Fresh samples were mixed with glycerol and frozen, thawed and prepared for SEM and TEM ( GA, OsO4, epoxy). There are no obvious signs of either gram-pos or neg. Is there any stain for sections? Fresh samples are no longer available.
TIA
Gunnel Karlsson
^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ Gunnel Karlsson E-mail Gunnel.Karlsson-at-oorg2.lth.se Biomicroscopy Unit Tel +46 222 8229 Inorganic Chemistry 2 Fax +46 222 4012 Box 124 S-221 00 LUND, Sweden ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ This message was sent by Eudora with recycled electrons
Hi Folks: We would like to do some standard processing and feature extraction from large images (about 10 megs, 8bit gray scale). Essentially we will use one image set to create a mask for feature extraction in another. The question is which packages are able to do this? I imagine the Photoshop image processing package (discussed a few weeks ago) will be able to generate the masks, however will this perform any quantitative extraction. Our current packages are limited by the frame size of the frame grabber, we would like to do this offline using standard ram memory. The platform can be PC, SGI or Mac.
Thanks
Simon C. Watkins Ph.D. Associate Professor and Director CBI University of Pittsburgh Pittsburgh PA 15261 tel:412-648-3051 fax:412-648-8330
Ultramicroscopy did exactly the same thing last year -- claiming "technical" problems then not appearing for many months. Yesterday I recieved my first/only copy of the year. I hope others will express (to elsevier) their frustration/anger at what used to be a good journal, but is now so unreliable as to be near useless. We are the customer, and the customer is right (or should be).
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University Evanston, IL 60208-3108 tel: (847) 491-3996 fax: (847) 491-7820 email: l-marks-at-nwu.edu http: //www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Has anyone encountered oil contamination problems on cooled TEM ccd cameras? We recently fitted a Gatan MSC 791 to the 35mm port of a Hitachi H7100 TEM and have encountered massive oil contamination on the cooled ccd YAG. Several solutions have been suggested: fit foreline traps to the RP pumps to minimise backstreaming (done), fit a cold trap to the camera chamber diff. pump (may not be enough space), fit a turbo pump (cost issues), fit a cold trap inside the camera chamber (?) or modify the way the camera is run e.g. cooled in use, then immediately switched to warm cycle and leave at ambient when not in use. Any suggestions on the best way to resolve this problem would be much appreciated - also any general information on how others run cooled cameras would be useful - continuously cooled or only when in use, time intervals between heating cycles, camera temperatures, oils used etc........ Thanks in anticipation
Kevin Jennings
SmithKline Beecham Pharmaceuticals Harlow Essex U K
} 4:25 PM } Ultramicroscopy } } Does anyone know what's become of the Elsevier journal "Ultramicroscopy?" ...
snips ...
} Still working in the dark... } Jeff Fortner } Argonne National Laboratory } Chemical Technology Division } Argonne, IL 60440
I undertake a regular literature review, every 2 months. The last issue of Ultramicroscopy I saw was 65(3/4) October 96, and the last time I was in the library was early April.
I have found similar problems with a number of other journals in the microscopy area. Personally, I would suggest that if publishers can't supply journals on a regular basis, the journal becomes irrelevant. Surely, major objective of science and publication in journals is about communicating information - if a nominally monthly journal doesn't appear for 6 months, then perhaps, unless reasonable explanations are provided, it is time to withdraw papers, cancel subscriptions and request refunds?
} I am looking for a film scanner (digitizer for HRTEM films), which } has resolution better than 2000 pixels/inch and 12 or more bits } per pixel. I found one from Kodak's products, but it is too expensive } (it works for color film, too). Anyone who has suggestion please } email to panx-at-umich.edu } Thank you. } Dear Xiaoqing, If you are doing quantitative work, you should consider something like the Perkin-Elmer or Optronics. (There may be other spot-scanners, but I don't know what brands they are.) The P-E has a 5 micron square aperture setting, and the scanning densitometers are *much* more accurate--especially with negs having large dynamic range over small distances. Stray light landing on a particular pixel in a CCD array can be significant if the pixel you're measuring is very dark, and this problem increases the nearer the bright pixels are to the dark ones. The low-order spots in an ED pat- tern, for example, have OD's of up to ~4, and proper background subtraction can only be performed if these are measured accurately. For quantitative comparisons of images to simulations, you may need similar accuracy, and, although you might be able to fold a "scanner contrast transfer function" into your simulation, it is better to get the measurements right in the first place. We have a P-E, but I have no financial interest in any of this. Yours, Bill Tivol
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Reply to: RE} Ultramicroscopy
We have taken contact with the editor in chief about the 97 volumes of Ultramicroscopy , Pieter Kruit , and the situation is as follows: After changing to computer aided production the publisher , Elseviers had a delay of several months Due to this the normally planned five volumes of 96 were not all published in that year. In order to keep their obligations the last volumes of 96 will come out in may 97 (They all contain material that was submitted in 96) From the end of may the new 97 volumes will appear ; Dirk van dyck
A Gatan 694 camera attached to the bottom of the camera chamber of our dry-pumped Hitachi HF-2000 initially exhibited quite bad oil contamination. After a couple of times removing the camera and cleaning the surface of the detector with a gentle wash of freon, the problem basically disappeared. We are convinced that the problem was caused by wicking of the oil film between the two fiber optic plates. Perhaps there was an initial excess of oil used to couple the plates together. Since there is no significant source of oil contamination in our microscope, other than what might come from O-rings, this seemed to be the only explanation. Interestingly, a similar camera on our JEOL 4000EX, which has a DP on the camera chamber, has not had any contamination problems. I think that if you have backstreaming sufficient to cause the problem on the CCD, you will have other contamination problems also....
Larry
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Why such large images (3000x3000 pixels)? Just getting and storing the images would seem a tremendous challenge. We routinely work with 1 MB images in Visilog. At that large stage, I would wonder about doing a hybrid system where the whole image is not stored. Detect the particles in a live mode and analyze them live or store just the area for your features of interest. Of course, this only works well with random access to all parts of the field.
I began image analysis with a 64 KB computer running hardware and software from LeMont Scientific. All intensities were read live from our SEM. Only the measurements were stored. Later versions also worked off of stored images. I have also seen a package from the R.J.Lee Group (Monroeville, PA, your back yard) which took a somewhat similar approach. Much of the measurement was done live, but images were also stored for each feature.
Disclaimer, I have no financial interest in the above companies. I remain fascinated by their ingenuity.
At 07:54 AM 4/23/97 -0400, Simon Watkins wrote: } ------------------------------------------------------------------------ } } Hi Folks: } We would like to do some standard processing and feature extraction from } large images (about 10 megs, 8bit gray scale). Essentially we will use one } image set to create a mask for feature extraction in another. The question } is which packages are able to do this? I imagine the Photoshop image } processing package (discussed a few weeks ago) will be able to generate the } masks, however will this perform any quantitative extraction. Our current } packages are limited by the frame size of the frame grabber, we would like } to do this offline using standard ram memory. The platform can be PC, SGI } or Mac. } } Thanks } } } Simon C. Watkins Ph.D. } Associate Professor and Director CBI } University of Pittsburgh } Pittsburgh PA 15261 } tel:412-648-3051 } fax:412-648-8330 ---------------------------------------------------- Warren E. Straszheim 270 Metals Development, Ames Lab/ISU, Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-3091
} I have a colleague who would like to know if there is a way to deside if } bacteria in activated sludge are gram-positive or gram-negative. Fresh } samples were mixed with glycerol and frozen, thawed and prepared for SEM } and TEM ( GA, OsO4, epoxy). There are no obvious signs of either gram-pos } or neg. Is there any stain for sections?
Gram negative can readily be distinguished from gram positive bacteria by examination of the structure of the cell wall by TEM. Check any good general bacteriology book for figures. The outermost layers ("wall") of gram negs show what appear to be two unit membranes with a thin (usually dense) amorphous layer between. This outermost membrane (containing lipopolysaccharide -} common in G- but extremely rare in G+) is most often undulated, giving the appearance of a ruffled surface by SEM. Gram positives, on the other hand, have a single cell membrane with a thick (usually electron-light) wall exterior to the membrane. The wall is composed of peptidoglycan and lacks LPS or lipopolysaccharide. If you need more info or require a specific reference, contact me.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 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 ####################################################################
} Has anyone encountered oil contamination problems on cooled TEM ccd } cameras? We } recently fitted a Gatan MSC 791 to the 35mm port of a Hitachi H7100 TEM and } have encountered massive oil contamination on the cooled ccd YAG. Several } solutions have been suggested: fit foreline traps to the RP pumps to minimise } backstreaming (done), fit a cold trap to the camera chamber diff. pump } (may not } be enough space), fit a turbo pump (cost issues), fit a cold trap inside the } camera chamber (?) or modify the way the camera is run e.g. cooled in use, } then } immediately switched to warm cycle and leave at ambient when not in use. Any } suggestions on the best way to resolve this problem would be much } appreciated - } also any general information on how others run cooled cameras would be } useful - } continuously cooled or only when in use, time intervals between heating } cycles, } camera temperatures, oils used etc........
Sounds like you have a vacuum problem with the EM. You should NOT be seeing "massive" oil contamination in this generation of microscope. Call the EM service people because the oil is going elsewhere in the EM as well.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 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 ####################################################################
Greetings collective, and special thanks to Nestor for clearing up those pesky double posts..
I'm working up an experiment that will compare freeze drying and freeze substitution on some Geranium pedicel trichomes. I would like to embedd in GMA and photo polymerize at a cool temperature, probably ca. 5C. After sectioning, I will de-embed and probe the sections with our imaging mass spect instrument (TOF-SIMS) to see how the chemical constituents have fared.
This is my first attempt with a GMA kit (EMS) and cold embedding/polymerizattion. Any tips from seasoned users.....? TIA
Edward J. Basgall, PhD The Pennsylvania State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618
Up to the age of forty, eating is beneficial; after forty, drinking. - The Talmud
TO WHOM IT MAY CONCERN, I heard that I could get some answers to my preperation problems here. I have been trying to prepare sickle cells for the SEM and I keep getting artifacts like echinocytes and folded cells. I would like to know if anyone could send me some special preperation instructions that I could use. Any help at all would be greatly appreciated. Thank you Sincerely, Emmanueuel Uche
I would like to announce an opening for - Senior Analyst/Microscipist; as of July 1, 1997.
An immediate position is open for a senior analyst at the North Carolina State University Analytical Instrumentation Facility (AIF) as a retirement replacement.
Duties and responsibilities include: operation and maintenance of optical metallographs, ion millers, X-Ray diffractometers and sample preparation devices such as mounting presses and grinding, polishing and sectioning devices, etc; scheduling of access to and oversight of the above instrumentation; and user training and assistance. Other responsibilities include operation of SEM and TEM and assistance with the teaching of electron microscopy laboratory classes and assistance with other graduate level engineering classes. Qualifications must include an BS as the minimum degree with higher degree desired or equivalent experience in a materials related discipline (non biological) along with hands on analytical experience in a multiuser analytical facility environment,
Please send resume and names of references to: Phil Russell, Director; Analytical Instrumentation Facility; North Carolina State University; Box 7531, Room 118A EGRC; 1020 Main Campus Drive; Raleigh, NC 27695-7531 (e-mail is acceptable; to prussell-at-ncsu.edu)
North Carolina State University is an Equal Opportunity, Affirmative Action Educator and Employer. Minority and Female Applicants are especially encouraged.
Phillip E. Russell Analytical Instrumentation Facility Box 7531, Room 318 EGRC North Carolina State University Raleigh, NC 27695-7531
A thread similar to this has been archive at the "Tips & Tricks " site. Go to the web address at the end of this message and look for the Tips & Tricks link. Go to the TEM section or use the search utility and you will find the replies to the earlier posting. Good luck
At 10:05 AM 4/23/97 +0200, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 352-392-1295 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
OOPS, sorry folks. The kit I will use is a methyl methacrylate/butyl methacrylate kit not GMA as I posted earlier.....
} I'm working up an experiment that will compare freeze drying and freeze } substitution on some } Geranium pedicel trichomes. I would like to embed } in MBM and photo polymerize at a cool } temperature, probably ca. 5C. } After sectioning, I will de-embed and probe the sections with our } imaging } mass spect instrument (TOF-SIMS) to see how the chemical constituents have } fared.
} This is my first attempt with a MBM kit (EMS) and cold } embedding/polymerizattion. Any tips } from seasoned users.....? } TIA
Edward J. Basgall, PhD The Pennsylvania State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618
If you really want a powerful, not necessary user friendly image processing package. I suggest you try semper6. Basically it has no limits of the image size. I use it to analysis images in the lab. Feel it is the one I most liked. Semper6 is a script language package. Not for dummy but for scientists. It has SGI, SUN and PC's versions. The problem is, it is not cheap. But you can get a demo version to try with.
I have no association with Semper6. It is developed by Cambridge U. (?)of UK.
I'm working up an experiment that will compare freeze drying and freeze substitution on some Geranium pedicel trichomes. I would like to embed in MBM and photo polymerize at a cool temperature, probably ca. 5C. After sectioning, I will de-embed and probe the sections with our imaging mass spect instrument (TOF-SIMS) to see how the chemical constituents have fared.
This is my first attempt with a MBM kit (EMS) and cold embedding/polymerizattion. Any tips from seasoned users.....? TIA
Edward J. Basgall, PhD The Pennsylvania State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618
We have a new Kevex Sigma 3 system with the Mirage image analysis software. Is there anyone out in EM land with a step-by-step protocol/instructions for imaging and analysis on Mirage? We are having difficulty deciphering the manual. Replies can be emailed to me or better yet, the instructions FAXed to me at 515.294.1337. My phone number is 515.294.3872. Thank you in advance!! Bruce Wagner.
We have the same system you are asking about and I had similar problems. If you think the Mirage manual is bad, you should have seen the manual for the Visilog 4 system I started with. The best advice I can give you is to read John Russ's "The Image Processing Handbook" or some similar text. If there is anything specific I can help you with, give me a call or send me an email.
John Robinson Electron Optics Technologist Mechanical & Materials Engineering University of Windsor Windsor, Ontario N9B 3P4
Hello, Lookng for the voice(s) of experienced TEM users (as opposed to vendors). What is a reasonable expectation for stage stability in a late model 200KV TEM. say in nm/min.?
I am interested in any hint in order to perform the calibration of a TEM high temperature sample holder test, up to 1000 degrees C. The idea is looking at various sample with a known structural transformation ocurring at precise temperature, and compare this "real" temperature with the one given by the sample holder thermocouple. Does anyboby have performed such a test before, and with which materials. I believe that having 4 or 5 points between 100 and 1000 degrees would be a good start.
thanks,
Yves MANIETTE Universitat de Barcelona Serveis Cientifico Tecnics Unitat ESCA TEM Carrer Lluis Sole i Sabaris, 1-3 E-08028 BARCELONA ESPANYA
Unfortunately, calibrating the electron-transparent portion of the TEM specimen to the TEM specimen holder temperature will be valid only for that particular specimen loaded at that particular time. Take the specimen out, and the former calibration will loose accuracy. This is due to the fact that you will not be able to duplicate the way in which you load (clamp) the specimen, so that the thermal contact resistance (between the specimen and specimen cup) will change. Fortunately, this likely amounts to only a few degrees of error. Furthermore, any calibration should only be used for similar specimens of similar thickness and having similar specimen preparation. If you calibrate the TEM holder using a specimen of relatively poor thermal conductivity, don't expect it to be correct for a specimen having high thermal conductivity. All bets are off if the specimen is not homogeneous in thermal conductivity (i.e., if the specimen has a crack in it, or is a multilayer, as the resistance to heat flow can vary locally so that the actual temperature can be significantly below the furnace temperature).
Its an interesting problem. We used the regrowth rate of amorphous silicon to establish the local specimen temperature in Si-Ge in:
D. C. Paine, D. J. Howard, N. D. Evans, D. W. Greve, M. Racanelli, and N. G. Stoffel, "In Situ TEM Studies of the Growth of Strained Si1-xGex by Solid Phase Epitaxy," in Evolution of Thin Film and Surface Microstructure, Mater. Res. Soc. Symp. Proc. 202, C. V. Thompson, J. Y. Tsao, and D. J. Srolovitz, eds., Materials Research Society, Pittsburgh, PA (1990)
} Date: Wed, 23 Apr 1997 12:49:15 -0700 (PDT) } From: Emmanuel Uche {euche-at-haywire.csuhayward.edu} } To: Microscopy Headquarters {microscopy-at-sparc5.microscopy.com} } Subject: NEED HELP W/ BLOOD SAMPLES } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } TO WHOM IT MAY CONCERN, } I heard that I could get some answers to my preperation problems } here. I have been trying to prepare sickle cells for the SEM and I keep } getting artifacts like echinocytes and folded cells. I would like to know } if anyone could send me some special preperation instructions that I could } use. Any help at all would be greatly appreciated. Thank you } Sincerely, } Emmanueuel Uche } A long time ago, we did some work on RBC's from muscular dystrophy patients. We found that the cells are very sensitive to pH, salt concentration, glass, and other things, maybe including the moon??? Low pH, increased salt, esp. NaCl, and sitting in a glass tube or on a glass slide unfixed would cause echinocytes. We got around the problem by drawing the blood and then IMMEDIATELY dropping (drop by drop) blood samples into 1% glutaraldehyde at pH 7.4, in 300 mOSM phosphate buffer, not PBS. Test the osmolarity of the buffer; the final osmolarity of the buffered fix will be 400: 300 from the buffer and 100 from the glut! The fix should be room temperature (not cold). Only about 2-3 drops of blood should be put into 10 ml fix. Gently invert several times to mix, but don't shake vigorously. Let them sit in the tube, and they will settle out to the bottom, or you can put a drop of suspension onto a collagen-coated coverslip, allow them to settle out for an hour (cover in a moist champer), then gently dehydrate in ethanol, and critical point dry. I don't remember which references contain the method, but try these:
Miller, S. E., A. D. Roses and S. H. Appel. 1975. Erythrocytes in human muscular dystrophy. Science 188:1131.
Roses, A. D., S. H. Appel, D. A. Butterfield, S. E. Miller and D. B. Chesnut. 1975. Specificity of biochemical and biophysical tests in Duchenne and myotonic muscular dystrophy, carrier states and congenital myotonia. Trans. Amer. Neurol. Assoc. 100:131-134.
Roses, A. D., M. J. Roses, S. E. Miller, K. J. Hull, Jr. and S. H. Appel. 1976. Carrier detection in Duchenne muscular dystrophy. New Engl. J. Med. 294:193-198.
Miller, S. E., A. D. Roses and S. H. Appel 1976. Scanning electron microscopy studies in muscular dystrophy. Arch. Neurol. 33:172-174.
Good luck, SM
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Dear list, We are looking for slides imprinted with grid squares so we can maintain orientation/coordinates. I was wondering if anyone has knowledge of any source that carries slides marked for orientation. Any help will be appreciated. with regards,
************************* * Marilyn Wadsworth * * mwadswor-at-zoo.uvm.edu * *************************
I am still having problems with my cpd efforts giving me 'raisins' especially on young plant tissue. Has anyone used HMDS? How do the results compare to CPD, or Freon (I know I can't use freon, but it used to be the standard)?
Are there any tricks I should know before embarking on a HMDS experiment?
Thanks for letting me pick your brains.
Paula = )
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
} Lookng for the voice(s) of experienced TEM users (as opposed to vendors). } What is a reasonable expectation for stage stability in a late model 200KV } TEM. say in nm/min.? } Dear Bruce, Not quite the answer to your question, but... The HVEM has a long- term drift of a few nm/min--I didn't have the time to look up our test re- sults. We test by inserting a stage, letting everything sit for ~1/2 hour and checking at 15 min intervals to see how far the image has moved. We found remarkably little movement until we looked more closely. It turned out that our Haskris chillers cycled with a period close to the observation interval, and when we looked continuously, we saw a cyclic drift pattern. We ordered a hot-gas-bypass unit for the chillers, and this gave much more stability. If your stability results are } } a few nm/min, you might look at some of your ancillary equipment (including room heating & air condi- tioning, etc.) to see if there is anything outside the scope causing in- stabilities. Good luck. Yours, Bill Tivol
This is a REPOSTING of the OFFICIAL PROPOSAL for a NEW USENET NEWSGROUP that will probably be named "sci.bio.immunocytochem" formally posted to news.announce.newgroups on 16.4.97.
Immunocytochemists/immunohistochemists may like to read the following proposal and post their comments or suggestions to "news.groups" where the set-up discussion is taking place, or you may e-mail me with your comments and I will repost them for you {awilson-at-aw.u-net.com}
REQUEST FOR DISCUSSION (RFD) unmoderated group sci.bio.immunocytochem
This is the 3rd Request For Discussion (RFD) for the creation of a world-wide unmoderated Usenet newsgroup sci.bio.immunocytochem, currently being discussed in news.groups
Suggestions for improvements to this proposal are welcome. Discussion about it should take place in news.groups. A vote is expected to be held in about three weeks.
This is not a Call for Votes (CFV); you cannot vote at this time. Procedural details are below.
CHANGES from previous RFD:
There is an important change to the Charter of this newsgroup: It is proposed that the new group will include the discussion of related affinity labelling methods as well as immunohistochemical and immunocytochemical topics. Minor changes to rational too.
Newsgroup line: sci.bio.immunocytochem Immuno-labelling of biological material.
RATIONALE: sci.bio.immunocytochem
Immunohistochemists and immunocytochemists already enjoy the benefits of online communication, utilizing e-mail, accessing web sites, and subscribing to specialised mailing lists. Usenet newsgroups are also popular, but this is less obvious because articles with immunocytochemical/immunohistochemical content get posted to many different newsgroups. Most articles are posted to a favourite five or six newsgroups including bionet.cellbiol, sci.med.immunology and sci.techniques.microscopy, but often articles get posted to any one of fourteen or fifteen newsgroups in the sci. and bionet. heirarchies. Some of these are listed in the distribution list at the end of this proposal.
In my view, no existing newsgroup fulfils the criteria necessary to attract all the various immunocytochemistry postings. I do not wish to draw users away from other newsgroups, only to encourage scientists to share their knowledge and expertise on immunocytochemistry in the most effective manner.
Immunocytochemistry and immunohistochemistry are not subdivisions of immunology, molecular biology or chemistry. Microscopy, although essential, is only a small part of the story. Immunocytochemistry and immunohistochemistry are multi- disciplinary, therefore discussions are destined to stay distributed amongst the different newsgroups until they are all brought together under one umbrella. This would then act as a focus point for all the immunocytochemists who are already Internet users, and encourage new subscribers to Usenet.
CHARTER: sci.bio.immunocytochem
This is a newsgroup for the exchange of information relating to immunocytochemistry and immunohistochemistry, and all forms of related affinity labelling methods, such as lectins and in-situ hybridisation. These unique research tools are used to locate and identify specific molecules in biological material, at the microscopical level.
Articles posted to this group must be relevant to one or more aspects of the above. The kind of subjects that may be discussed include techniques, theory, presentation of results, requests for collaboration, history, equipment, publication references, notice of events, tips and trouble-shooting, jobs offered andwanted, jokes, stories and new ideas, so long as the posting bears a direct relevance to the central theme. There will be a list of Frequently Asked Questions (FAQs) to help newcomers.
A relevant posting could just be a simple question or answer, for example "Has anyone got any experience with this reagent ?"or "Which course could I attend to learn more about immunogold labelling?". There will be articles reminding people to read the list of FAQs prior to posting their own article. Usenet readers may get involved in complex discussions about, for example, multiple labelling, proper use of control experiments, microwave antigen retrieval or quantitative measurements. Remember that articles posted to a newsgroup are intended for a wide readership, so if you have information which concerns only one or two people then please don't use this newsgroup, use e-mail.
Commercial advertisements for services, equipment or reagents violate the charter unless one or more of the following apply: (a)The advertisement is part of a comprehensive article designed specifically to address issues raised in earlier articles posted to the group (b)A general reference to the type of product does not suffice for technical reasons and it is necessary to specify the exact commercial product (c) The information is offered primarily for the benefit of the readers (d)The advertisement is for second-hand equipment specific to immunocytochemistry (e) Requests or offers for free products are acceptable if they are not part of a sales promotion.
END CHARTER.
PROCEDURE:
This is a request for discussion, not a call for votes. In this phase of the process, any potential problems with the proposed newsgroups should be raised and resolved. The discussion period will continue for a minimum of 21 days (starting from when the 3rd RFD for this proposal is posted to news.announce.newgroups), after which a Call For Votes (CFV) may be posted by a neutral vote taker if the discussion warrants it. Please do not attempt to vote until this happens.
All discussion of this proposal should be posted to news.groups. This RFD attempts to comply fully with the Usenet newsgroup creation guidelines outlined in "How to Create a New Usenet Newsgroup" and "How to Format and Submit a New Group Proposal". Please refer to these documents (available in news.announce.newgroups) if you have any questions about the process.
DISTRIBUTION:
This RFD has been posted to the following newsgroups: news.announce.newgroups,news.groups,bionet.cellbiol, bionet.diagnostics,bionet.immunology,bionet.microbiology, bionet.molbio.methds-reagnts,sci.bio.misc,sci.med.immunology, sci.techniques.microscopy
This RFD will be reposted to the following newsgroups after its posting in news.announce.newgroups: bionet.molbio.proteins,bionet.neuroscience,bionet.plants, sci.bio.microbiology,sci.med,sci.med.laboratory,sci.misc, sci.nanotech
This RFD will also be reposted to the following mailing lists after its posting in news.announce.newgroups:
Histonet mailing list: {histonet-at-pathology.swmed.edu} Information pertaining to the technical aspects of histology and histopathology such as tissue fixatives and processing, routine histology, special stains, immunohistochemistry, in-situ hybridization etc. To subscribe type "subscribe digest" into the subject box and leave the text box empty, or to subscribe to the full service just type "subscribe". For more info access web site http://www.mwrn.com/subject/histonet.htms
Microscopy Society of America listserver: Questions/comments/answers in the various fields of Microscopy Currently over 3000 subscribers. To subscribe send the message "subscribe" to {Listserver-at-MSA.Microscopy.Com} then send messages in plain text to {Microscopy-at-MSA.Microscopy.Com} For more info access web site http://www.amc.anl.gov/ Docs/anl/Nestor/Software/telecommList.html
Stanford University list server To subscribe, send a message to {majordomo-at-pathology.stanford.edu} with "subscribe ipox-l" in the body of your message. This list helps pathologists and other laboratory professionals to exchange information about immunoperoxidase methods.
This RFD will also be reposted to the following web-sites after its posting in news.announce.newgroups:
Royal Microscope Society http://www.rms.org.uk Web Master Dr R. A. D. Mackenzie {r.a.mackenzie-at-open.ac.uk}
Center for Cell Imaging Department of Cell Biology Yale University School of Medicine Introduction to Immunocytochemistry http://info.med.yale.edu/cellimg/CCIimmuno.html Web Master Paul Webster { paul_webster-at-yale.edu}
Proponent: Amanda Wilson {awilson-at-aw.u-net.com} Proponent: Paul Monaghan { monaghan-at-icr.ac.uk} Mentor: Jonathan Grobe {grobe-at-netins.net}
To all those folks who have a violent interest in Pb stains for TEM, and to all those who have requested a copy of the book chapter -
I am sorry to have have had to delay my answers to you all on this most intriguing topic - we have been suffering here from MEGA grant insanity. On Tuesday, April 29, I will plan to post a message and bundle up the book chapter copies. Thanks for your patience. Hildy
Hi, Although I guess I could be considered a vendor; I have been known to use a microscope from time to time... The question you ask is not complete; I could reply that any microscope stage which does not use sliding 'O' rings can be seen to have drift rates well below 1 nm/min, depending on how much you are prepared to spend on the site. It is very much a factor of the temperature stability of the sample in the holder, the holder/sample temperature, and the sample temperature when inserted should be as equal to the in-column temperature as possible to give the best results. When you have all the thermal factors pretty well sorted out (now you've spent the best part of $100,000 on THE ROOM!!) i.e. no drafts and no difference in temperature between the sample whether it is inside the column or outside the column, THEN (and only then) can you start to think about what happens when you turn on the beam. IF your sample conducts heat reasonably, then you have a chance at good stability below 3 angstroms/min. There are not many vendors who would endorse such numbers because of the site variables that are involved. If the sample has poor conductivity, then the beam will cause local heating and then the sample will move no matter what you try to do.
Any other inputs on this subject would be much appreciated. I know that Tim Baker at Purdue has measured some unbelievable drift rates using a cold stage (COLD!) but I cannot quote them here...
Cheers, Jan
Dr. Jan Ringnalda Sr. Apllications Specialist, FEI/Philips Electron Optics, 85 McKee Dr., Mahwah, NJ 07430 (201) 529 6160
Dear Bruce, From my own experience, the sample should drift several nm /min for the first 1/2 hour, then settle down to little or no drift after that, unless the specimen itself moves under the beam. Speaking of voices, if you observe the image at ~200,000 and talk to someone, you will see the specimen vibrate when your voice hits certain frequencies. Remember not to talk while you are photographing. A microscope designer told me they once improved stage stability by reducing the diameter of the handle of the specimen holder, so it didn't react to air drafts and thermal gradient so much. This is why high resolution TEMs are top entry. Air drafts, air conditioners, noisy water chillers, any other noise or source of drafts or thermal changes will affect the stage. You wrote:
} Hello, } Lookng for the voice(s) of experienced TEM users (as opposed to vendors). } What is a reasonable expectation for stage stability in a late model 200KV } TEM. say in nm/min.?
Hope this helps, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
} We would like to do some standard processing and feature extraction from } large images (about 10 megs, 8bit gray scale). Essentially we will use one } image set to create a mask for feature extraction in another. The question } is which packages are able to do this? I imagine the Photoshop image } processing package (discussed a few weeks ago) will be able to generate the } masks, however will this perform any quantitative extraction. Our current } packages are limited by the frame size of the frame grabber, we would like } to do this offline using standard ram memory. The platform can be PC, SGI } or Mac.
I use Khoros2, which comes as a shareware 'Advanced' version and a commercial 'Pro' version (I think the differences are minor, but I only know the 'Advanced'). It runs on any computer running some sort of Unix (Sun, SGI, PC ...). I've processed byte images up to 4096 pixels on a Sun with 96 MByte RAM. Khoros will do almost anything You can imagine (or at least I can) in the field of general image processing. Check: http://www.khoral.com/
Hope that helps, -- Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 86 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www_scem.csb.ki.se/pages/philip.html
} I think Larry might be right in his case but that John B. is probably right } in your case. There is no way to tell, though, without knowing more about } the extent and duration or your problem. Could you elaborate on your } problem and include camera serial number and date shipped (if known)?
} Paul
Thanks to everyone for the comments so far. Here are more details - First noticed the problem after a couple of weeks use - overnight heating appeared to remove the contamination but it continued to reappear in the same position by the end of the day with continuous cooling. The camera (serial number K6070403 delivered Jan 97) was removed by an engineer after one of the heating cycles and found to be contaminated with oil in the same position as seen on captured images. After cleaning, the camera was reassembled and all appeared OK - although on switching on the following morning the camera failed completely with no image. On checking the camera, more oil was found plus the whole ccd /Peltier assembly appeared loose. This should be fixed next week after which we will review the problem again. As for microscope oil problems - the H7100 appears to have low contamination rates in normal use and has given no obvious problems whilst we've been using film.
Kevin Jennings
SmithKline Beecham Pharmaceuticals Harlow Essex CM19 5AW U K
} I am interested in any hint in order to perform the calibration of a TEM } high temperature sample holder test, up to 1000 degrees C. The idea is } looking at various sample with a known structural transformation ocurring } at precise temperature, and compare this "real" temperature with the one } given by the sample holder thermocouple. Does anyboby have performed such } a test before, and with which materials. I believe that having 4 or 5 } points between 100 and 1000 degrees would be a good start. } } thanks, } } Yves MANIETTE } Universitat de Barcelona
While this sounds like a good approach, I'm not sure it would give you a better accuracy than you already have with the built-in thermocouple. I think a lot depends on how accurately you really do need to know the temperature? You might also have difficulties finding a suitable material (that is, you can make a TEM specimen) at the lower end of the temperature range.
Firstly, the temperature given for such transformations probably applies to bulk material. It is possible that in very thin TEM specimens, the transformation temperature may be different?
Secondly, in most cases, such transformations aren't instantaneous - they move through a material from nucleation sites. As a consequence, I would expect the transformation to occur over a small temperature range, rather than at a specific temperature. Of course, if the range is small, say 1 degree, this may be sufficient for your purposes.
Thirdly, the thermocouple is measuring the temperature not of the specimen, but of some point near to the specimen, either in the holder tip, or, more likely, in the specimen cup. Thermal conductivity and specimen clamping considerations mean that there will always be a thermal lag between the specimen and the thermocouple; there may also be a temperature gradient, although I guess that in most cases this would disappear given a suitable period of time. This effect may also be influenced by beam heating, and will be different for every specimen.
An alternative approach (which I have not personally tried, so I don't know if it would really work) may be to measure thermal expansion. Although electron diffraction is not too good at making an absolute measurement of lattice parameter, convergent beam methods, looking at HOLZ lines can in principle measure lattice parameter changes to 1 in 1000 or 10000. If you start with a material with precisely known room temperature lattice parameter and thermal expansion coefficients, you should be able to measure the lattice parameter at several temperatures and then calculate back to a value for the actual temperature. This approach has the advantage that you really are measuring the temperature of the specimen plus you can decide at which temperature points to calibrate, and how many. It might also allow you to do such a calibration in situ - that is, with the material in which you are really interested.
I would suggest you contact both Gatan and Oxford Instruments, who make commercial TEM heating holders. I don't know about Oxford, but at Gatan contact Reza Alani {ralani-at-gatan.com} , their Applications Manager.
It's not really a drift problem, but I recall several years ago that I used a Philips 301(ironically) with eucentric goniometer and the stage seemed to have a resonant frequency somewhere near human voice levels - at least that's what we thought. This meant that the best pictures were taken by the quietest people. I don't know if similar problems still arise but they could obviously make normal specimen drift difficult to standardise.
Malcolm Haswell University of Sunderland UK ----------
Hi, Although I guess I could be considered a vendor; I have been known to use a microscope from time to time... The question you ask is not complete; I could reply that any microscope stage which does not use sliding 'O' rings can be seen to have drift rates well below 1 nm/min, depending on how much you are prepared to spend on the site. It is very much a factor of the temperature stability of the sample in the holder, the holder/sample temperature, and the sample temperature when inserted should be as equal to the in-column temperature as possible to give the best results. When you have all the thermal factors pretty well sorted out (now you've spent the best part of $100,000 on THE ROOM!!) i.e. no drafts and no difference in temperature between the sample whether it is inside the column or outside the column, THEN (and only then) can you start to think about what happens when you turn on the beam. IF your sample conducts heat reasonably, then you have a chance at good stability below 3 angstroms/min. There are not many vendors who would endorse such numbers because of the site variables that are involved. If the sample has poor conductivity, then the beam will cause local heating and then the sample will move no matter what you try to do.
Any other inputs on this subject would be much appreciated. I know that Tim Baker at Purdue has measured some unbelievable drift rates using a cold stage (COLD!) but I cannot quote them here...
Cheers, Jan
Dr. Jan Ringnalda Sr. Apllications Specialist, FEI/Philips Electron Optics, 85 McKee Dr., Mahwah, NJ 07430 (201) 529 6160
I am looking for a secondary detector tube for JEOL JSM 35 in a good condition. Willing to pay a fair price for it. Please reply at my email address. Many many thanks for your anticipated co-operation.
Jitu Shah
Dr.Jitu Shah H.H. Wills Physics Laboratory, University of Bristol, Royal Fort, Tyndall Avenue, Bristol BS8 1TL. UK email: jss-at-siva.bristol.ac.uk Tel: 44 117 9288719 Fax: 44 117 9255624
Generally, HMDS does not work on plant tissue. During the SEM course we teach, we have made the students use both procedures ( we are such slave drivers) and there seems to be no way to tell which plants it will work on and which it won't. Have you tried long infiltrations??? Atr the end of the CPD run do you allow the gas bleed off slowly?? It usually takes us 20-30 min. to come to atm.
I maintain an archive of the biologic related threads posted to this list. You may find it at the web address listed at the end of this message. Go to the "Tips & Tricks" link and look in the "SEM" section. There are some threads you may find enlightening. Good luck!!
At 01:45 PM 4/24/97 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 352-392-1295 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
We have never had good luck with HMDS on plant material. We always CPD. Be sure that ou have gotten ridof all the water and solvent before drying Sniff the chamber when you open and see if you still smell solvent. If yes you need longer or more changes during CPD } } Dear smart people, } } } I am still having problems with my cpd efforts giving me 'raisins' } especially on young plant tissue. Has anyone used HMDS? How do the } results compare to CPD, or Freon (I know I can't use freon, but it used to } be the standard)? } } Are there any tricks I should know before embarking on a HMDS experiment? } } Thanks for letting me pick your brains. } } } Paula = ) } } Paula Sicurello } UC Berkeley } Electron Microscope Lab } psic-at-uclink4.berkeley.edu } } } } ******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Marilyn Wadsworth writes:
"We are looking for slides imprinted with grid squares so we can maintain orientation/coordinates. I was wondering if anyone has knowledge of any source that carries slides marked for orientation. Any help will be appreciated."
Dear Marilyn,
I do not know of glass slides that are etched but what you may be looking for are coverslips with etched grids.
You may wish to contact "Eppendorf" to ask about their "CELLocate" system. You get small glass, sterile coverslips, etched with a numbered grid. Paper maps are supplied to mark interesting locations.
Cells grow well on the substrate and the etched grid will transfer easily to epoxy resin after embedding.
Regards,
Paul Webster Center for Cell Imaging Yale School of Medicine http://info.med.yale.edu/cellimg
The Electron Microscopy Laboratory at New Mexico State University has an open position for an Electron Microscopy Specialist. The laboratory provides transmission, scanning and light microscopy services for the university research community. Qualifications: B.Sc. degree minimum, M.Sc. degree desirable, with at least 4 years of electron microscopy experience. The preferred candidate will have experience with scanning electron microscopy and x-ray analysis. Experience with digial image analysis is also desirable as is experience with flourescense microscopy and immunocytochemistry. The candidate must be competent with sample preparation techniques including vacuum evaporation, sputter coating, critical point drying, support film production, low temperature embedding, and photographic film processing and printing. He or she must be able to work well with research faculty, staff and students, and be able to train and instruct graduate and undergraduate students. Duties: operation and routine maintenance of transmission and scanning electron microscopes and associated equipment, fixation, embedding, ultra-thin sectioning, staining, coating of samples, and record keeping. Salary: $26,872 to $30,972 plus a 26% fringe benefits add on. Applications will be accepted beginning May 1and will continue until the position is filled. Send resume and 3 letters of recommendations to: Director, Arts and Science Research Center Box RC, New Mexico State University Las Cruces, NM 88003
Hank Adams Electron Microscopy Laboratory NMSU 505-646 3600
I seem to have reached the zenith of my very limited knowledge of immunohistochemistry. I would be grateful for any assistance you can give on the following problem:
I am trying to use a fish monoclonal anitbody to vitellogenin (a yolk protein precursor) to stain fish gonad and liver 7u parafin sections. On vitellogenic oocytes I am seeing what appears to be good specific staining. The problem is that I also see the same intensity of staining with secondary only, thus my problem (TRITC goat anti-mouse; I tried FITC but got a lot of autoflouresence). I am blocking with 1% BSA 10 min. Primary incubation is overnight at room temp, concentration of 25 ug/mL; secondary is 30 min. concentration of 1:50 (also tried 1:200 same response).
Diana Papoulias Fish Biologist, Research Midwest Science Center Biological Resource Division U.S. Geological Survey 4200 New Haven Rd. Columbia, MO 65201 Tel 573 875 5399 xt 1902 Fax 573 876 1896 email: Diana_Papoulias-at-nbs.gov
Hi Paula, Dr. Beverley Giamarra has done a lot of work with HMDS. Below is a bibliography of HMDS work I am familiar with: If you hear of any others I'd appreciate your forwading them to me.
HMDS
Adams JL, Battjes DA, and Buthala DA. (1987). Biological Specimen Preparation for SEM by a Method Other Than Critical Point Drying. Proc. EMSA. 45, 956-957.
Giammara B, Baker R, Burkes E, Malangoni M, Evers B, and Hanker J. (1987a). Hexamethyldisilazane for Rapid Scanning Electron Microscopic Examination of Implant Specimens. (abs) J. Dent. Res. 66, 187.
Giammara B, DeVries W, Baker R, Dobbins J, and Hanker J. (1987b). Hexamethyldisilazane Drying for Rapid Detection of Bacteria in Implant Specimens. Proc. EMSA. 45, 878-879.
Giammara B and Hanker J. (1988a). Ruthenium Red-Osmium Bridging with TCH: New Technique to Stain Biological Specimens for Light and TEM and to Coat for SEM. Proc. EMSA. 46, 20-21.
Giammara BL and Hanker JS. (1988b). New Ruthenium Red Bridging Technique with Thiocarbohydrazide to Stain Polyanionic Biomacromolecules for Light and Electron Microscopy and to Coat Biological Specimens for Scanning Electron Microscopy. Proc. 9th Eur. Cong. Elec. Mic.
Giammara B, Washburn M, Malangoni M, Evers B, Baker R, Burkes EJ, and Hanker J. (1987c). Rapid Scanning Electron Microscopic Examination of Implant Specimens with Hexamethyldisilazane Drying. Soc. for Biomaterials Trans. X, 103.
Lamoreaux W. (1988). Prevention of Outgassing When Coating Tissues Dried with Hexamethyldisilazane (HMDS). EMSA Bull. 18, 91.
Nation JL. (1983). A New Method Using Hexamethyldisilazane for Preparation of Soft Insect Tissue for Scanning Electron Microscopy. Stain Technol. 58, 347-351.
HMDS SAFETY NOTES HMDS is flammable, use in a flammables rated exhaust hood. Avoid breathing vapors. It is toxic by inhalation, toxic sin contact with skin, or if swallowed. I have observed that HMDS seems to produce an ammonia compound when mixed with ethanol.
Other than the above common sense precautions, it is very easy to use as a final solvent treatment after fixation, post-fixation and dehydration. I have not used it myself with plant material.
good luck
Edward J. Basgall, PhD The Pennsylvania State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618 } } Dear smart people, } } } I am still having problems with my cpd efforts giving me 'raisins' } especially on young plant tissue. Has anyone used HMDS? How do the } results compare to CPD, or Freon (I know I can't use freon, but it used to } be the standard)? } } Are there any tricks I should know before embarking on a HMDS experiment? } } Thanks for letting me pick your brains. } } } Paula = ) } } Paula Sicurello } UC Berkeley } Electron Microscope Lab } psic-at-uclink4.berkeley.edu
Marylin, From your message it wasn't clear to me how small your grids need to be. But here are a few suggestions anyway. Bellco Glass sells a coverslip with an etched grid pattern. I've used these very successfully for light microscopy e.g. locating cell which I have microinjected. I have also made my own indicator coverslips by using a locator EM grid as a mask and evaporating a layer of carbon (LM only) or gold (LM or EM). The gold layer need not be thick. The problems with metal evap are that (1) you must clean and bake the glass before and after metal deposition otherwise it might lift off when you place it in media or buffer (2) the metal will act as a neutral density filter and attenuate light, it may even burn up if you are using a laser for imaging (depends on incident intensity). You can avoid this problem by just working with cells off the metal. Hope these suggestions are helpful. If you have any other questions don't hesitate to contact me. Victoria Centonze Frohlich Deputy Director, IMR University of Wisconsin, Madison Symposium ($30) and Workshop ($230) on "Applications of Multiphoton Excitation Imaging" August 9 and 10, 1997 Sheraton City Center, Cleveland, OH To Register Contact: dvolkman-at-students.wisc.edu www.bocklabs.wisc.edu/imr.html
} We are looking for slides imprinted with grid squares so we can maintain } orientation/coordinates. I was wondering if anyone has knowledge of any } source that carries slides marked for orientation. Any help will be } appreciated.
I am interested in finding out some of the ways that people tend to quantify retroviral particles (as a whole, not just infectious). We have been using a vendor that performs thin-section TEM and have looked some into negative stain TEM. We have not seen consistent results from the thin section determinations, and have been working on the sample prep end to get a more consistent pellet volume and consistency to submit for TEM.
I am interested in hearing what other people tend to do for retroviral quantitation (particularly in the Biotech industry because of FDA concerns). If I could get some input from this listserv group, I would appreciate it. The address above is correct. Thank you.
Do you have any protocols for EM in-situ using digoxygenin labelled probes? My samples are already embedded in LR White, is this OK for that? These probes work really well for light microscopy in-situs.
Thanks for any information you can give me on this subject.
Paula = )
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
From my field experience - indeed most part of stage drift in HVTEM comes from thermal influences from cooling systems. Once investigating such extensive drift by CM-20 we measured very precisely obj. lens-water temperature and found out, that the image drift observed by second highest mangnification step follows exactly the water cooler regulation. In fact all coolers which have temp. level regulation mechanism with histeresis are not suitable for todays HVTEM. Philips f.e. recommends the units with follow-up temp. regulation so theoretically (and very close practically) such unit by constant operating conditions (constant power) can reach a steady state - constant water temperature. The ,,histeresis'' units can't. When experimenting with such water supply in a/m example we achieved finally - to have the same feature in the screen center (by second highest magn.) for 30 min without any noticable movement (!) (only waht was noticable - growing contamination ring). That was in very patient test conditions - it's not the case by normal work, ofcourse. regards Krzysztof M. Herman E.Eng M.Sc. Philips El.Opt. Service Specialist LabSoft S.c. 05-500 Piaseczno, 21 Kosciuszki Str. tel/fx: (48 22) 7502024, 7502028, 7570671 fax only: (48 22) 483787, Email: labsoft-at-ikp.atm.com.pl
I once again have called on the services of Bernie Kestel from Argonne National Laboratory and he has given me the following information:
"Selective etch of Cu from cobalt , non-electolytic. Try dilute nitric acid in methanol, NOT ethyl alcohol, (an explosive mixture). Water as a solvent may also work. Try 3-4 % acid & increase concentration if needed to get some action. Heat is generated when mixing the acid mixture, so cool with ice or dry ice before adding acid. Use at room temp. Cobalt is usually electropolished with perchloric acid mixtures so the nitric acid may not attack it much. In electropolishing, Cu works in nitric solutions while Co works in perchloric solutions. Even my non-acid bath, BK-2, does not work on Cu. I don't know how to thin Cu & Co at the same rate. Develop a new non-acid electrolyte, perhaps."
Bernie does his work with our Model 550 Single Vertical Jet Electropolisher, but other jet polishers will certainly work well too.
You can get additional information on the Jet Polisher (Now Model 550D) on our web site at http://www.southbaytech.com.
DISCLAIMER: As we do manufacture the Model 550D Single Vertical Jet Electropolisher, I obviously have a vested interest in promoting its use.
David Henriks TEL: 800-728-2233 (toll-free in USA) South Bay Technology, Inc. 714-492-2600 1120 Via Callejon FAX: 714-492-1499 San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
Dear Diana, Have you tried blocking with 10% goat serum? I also add 10% goat serum in my primary and secondary antibodies. Another point to consider is your source of secondary! Is the staining specific or non specific...does it stain up the same area of what you expect to get with the addition of primary? I would suggest you try some other tissues as positive and negative controls and if all else fails, maybe try another source of secondary antibody...or if possible, purify the secondary through an appropriate affinity column. Hope this helps!
K.M.Khoo, Department of Biochemistry, Faculty of Medicine, National University of Singapore.
On Thu, 24 Apr 1997, Diana Papoulias wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } List readers, } } I seem to have reached the zenith of my very limited knowledge of } immunohistochemistry. I would be grateful for any assistance you can } give on the following problem: } } I am trying to use a fish monoclonal anitbody to vitellogenin (a yolk } protein precursor) to stain fish gonad and liver 7u parafin sections. } On vitellogenic oocytes I am seeing what appears to be good specific } staining. The problem is that I also see the same intensity of } staining with secondary only, thus my problem (TRITC goat anti-mouse; } I tried FITC but got a lot of autoflouresence). I am blocking with 1% } BSA 10 min. Primary incubation is overnight at room temp, } concentration of 25 ug/mL; secondary is 30 min. concentration of 1:50 } (also tried 1:200 same response). } } } Diana Papoulias } Fish Biologist, Research } Midwest Science Center } Biological Resource Division } U.S. Geological Survey } 4200 New Haven Rd. } Columbia, MO 65201 } Tel 573 875 5399 xt 1902 } Fax 573 876 1896 } email: Diana_Papoulias-at-nbs.gov } }
This is THE FINAL URGENT REQUEST for your comments, suggestions, etc about the 3RD RFD for my PROPOSED NEW NEWSGROUP SPECIALIZING IN IMMUNOCYTOCHEMISTRY/IMMUNOHISTOCHEMISTRY/OTHER RELATED AFFININITY METHODS called "sci.bio.immunocytochem" now posted in "news.groups"
The latter contains material of a rather varied nature (!) but it is necessary to use this unmoderated group to hold the discussions about all the different proposed new groups.
So, if you are connected to Usenet, and you are keen to see A NEW IMMUNOCYTOCHEMISTRY NEWSGROUP, then please go to "news.groups", ignore all the rubbish, look for articles posted on 20.4.97 or 24.4.97 (or later) and you should find one of my postings "3RD RFD: sci.bio.immunocytochem". Select "follow-up article" (or equivalent) from your newsreader menu, and post your message so that it appears under the 3RD RFD.
If you don't have access to Usenet, you can read the proposal at the Royal Microscope Society web site {http://www.rms.org.uk} , or at the Introduction to Immunocytochemistry (Center for Cell Imaging Department of Cell Biology Yale University School of Medicine) web site {http://info.med.yale.edu/cellimg/CCIimmuno.html}
MANY THANKS to all of you who have already posted your responses to my RFDs! BUT WE STILL NEED MORE DISCUSSION in news.groups please! SOON I shall be posting my "CALL FOR VOTES" to the people in charge of Usenet newsgroups. When you see "CFV: sci.bio.immunocytochem", you will be able to e-mail your vote to the vote-taker.
Hi: I have received two E-mail letters from Philip Koeck over the past week or so through this forum. Both letters I can not read or delete. Whenever I try I get an error message saying that the program has performed an illigal action and turns netscape off. Anybody having the same problem or any solutions? Thank you, Peter Jordan
Peter Jordan wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hi: } I have received two E-mail letters from Philip Koeck over the past week } or so through this forum. Both letters I can not read or delete. } Whenever I try I get an error message saying that the program has } performed an illigal action and turns netscape off. Anybody having the } same problem or any solutions? Thank you, Peter Jordan
Yes I am. It's very frustrating. I'd appreciate any help.
Peter Jordan wrote: } } Hi: } I have received two E-mail letters from Philip Koeck over the past week } or so through this forum. Both letters I can not read or delete. } Whenever I try I get an error message saying that the program has } performed an illigal action and turns netscape off. Anybody having the } same problem or any solutions? Thank you, Peter Jordan .......................... I also use Netscape 3.0 and have the same problem. I don't have a solution, but I do have a way to get rid of the "poison message":
(1) read, and delete or transfer all the rest of the mail in the Inbox, but don't try to read or delete the "problem" message. (This is tricky, and you may have to "error-out" and reload Netscape a couple times.) (2) When the Inbox is empty except for this message, exit from Netscape. (3) Go to the NETSCAPE\NAVIGATOR\MAIL subdirectory, and locate the file "Inbox". Delete it (or rename it if you want to be cautious). (4) The next time you load Netscape, it will recreate an empty Inbox file.
This is ugly, but it's the only way I've found to get rid of the darn thing!
I hope somebody who doesn't have this problem will help us out by sending a message to the originator to find out what he is doing! I can't open his messages to get a return address. -- Fred Schamber ....................... mailto:fhscham-at-SGI.NET
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Frederick Schamber wrote:
I also use Netscape 3.0 and have the same problem. I don't have a solution, but I do have a way to get rid of the "poison message":
(1) read, and delete or transfer all the rest of the mail in the Inbox, but don't try to read or delete the "problem" message. (This is tricky, and you may have to "error-out" and reload Netscape a couple times.) (2) When the Inbox is empty except for this message, exit from Netscape. (3) Go to the NETSCAPE\NAVIGATOR\MAIL subdirectory, and locate the file "Inbox". Delete it (or rename it if you want to be cautious). (4) The next time you load Netscape, it will recreate an empty Inbox file.
This is ugly, but it's the only way I've found to get rid of the darn thing!
I use Netscape V. 3.01 and I have the same problem. This is the way I deal with it:
I discover the message that is causing trouble (sometimes you'll have to launch Netscape several times to find out the message). Then I mark a group of messages including the "problem" message (never mark it at first or last. Keep it among others). You'll have to do the same again when they are transferred to the trash directory. This procedure works well most of time.
I would like any information you have on the KEVEX 8000 upgrade system. I am specifically looking for how many channels can be imaged at once. i.e. number of EDS maps displayed at one time and how many external channels (WDS) can also be displayed with the EDS?
Also, how do you like the operation of the system?
The problem appears to be associated with Netscape Email readers as I can read and delete the file using other Email programs (i.e standard Mail on Unix and Eudora on PC or Mac Platforms). I have sent a message both to NETSCAPE and to Philip Koeck {Philip.Koeck-at-csb.ki.se} the originator of the message. I noticed the Koeck uses Netscape Gold V 3.01 on a Windows 95 Machine (you can find this out by reading his Email header file)
It would probably help for those of you running Netscape who have the problem to also report it to NETSCAPE. They have an on-line form for this at:
To all who responded to my plea for help: Thank you very much. Sandwiching the "bad" files between two "good" files allows you to move them into the trash bin and then to delete them. For all computer greenhorns like me this is done by clicking on the good file, then holding down the shift key clicking on the other good file. All files between the clicked files will be marked and then can be moved. It was realy nice to get all these respones. Thanx again, Peter Jordan
Hi: I forgot to give credit to Dan Kaszubski for the solution on how to delete Philip Koeck's E-mail letters. Thank you, Dan. I did not realize how many of us had this problem. The recipe is in my other letter. Peter Jordan
Dear all, I am aware of a number of programs that can produce calculated diffraction patterns given the unit cell parameters and the location of all atoms in the unit cell. This poses difficulties for some materials with more complex structures, however, where site occupancies are not 100% and where the 'unit cell' is somewhat of an average structure (this occurs for some oxides). This can be overcome if a good crystal structure refinement has been done from XRD where average atomic positions and site occupancies have been determined. Many structures have not been studied in this detail, however, and the only information that is available is the unit cell parameters and the space group. Is there any software that can plot electron diffraction patterns from this limited information.
I hope there is someone who can help me with this.
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
} } } } } } } } } } } } } } } } } } } } } .. } } I am a research scientist in Columbia University and I need some Quartz } Cover Slips badly for one of my projects. I was unable to locate any. } Would you please kindly send me a list of vendors who may carry this } product that I can buy from. } My email address is hl51-at-columbia.edu } I would appreciate it very much, } } Sinserely your, } } Hui Lao, M.D. } } ******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
Dear Marilyn, Following is information on the grided slide. FIELD FINDER Microslide Field Finder helps you index points of interest on microscope slide the same way you find them on a road map. Standard 75X25mm slide has photographically imprinted grid of 1mm squares, subdivided into 0.1mm intervals. eash square is marked with letter and number. You center detail in field of microscope...replace specimen slide with field finder...and read coordinates. The slide is provided by the following: Fisher Scientific 1-800-766-7000 Catalogue number# 12-454 Price each $142.00
I am not affliated with this company in any way. I am just passing on the information.
Dear list, We are looking for slides imprinted with grid squares so we can maintain orientation/coordinates. I was wondering if anyone has knowledge of any source that carries slides marked for orientation. Any help will be appreciated. with regards,
************************* * Marilyn Wadsworth * * mwadswor-at-zoo.uvm.edu * *************************
I am Electrical-Engineer , and i have in propiety Electron -microprobe i I would like any information you about electron -microprobe result of smectite Clinoptilolites and others Zeolites .
I think you will probably get lots of replys from the vendors for your question, but I thought I'd tell you a few of the companies that I have used for years with satisfactory results.
Electron Microscopy Sciences, 321 Morris Rd, Box 251, Ft. Washington, PA 19034, 1-800-523-5874
Ernest F. Fullam, Inc., 900 Albany Shaker Rd, Latham, NY 12110, (518)785-5533
Ted Pella, Inc., e-mail tedpel-at-aol.com or tedpel-at-snowcrest.net
These are just my own picks, I'm not affiliated with any of them.
Karen Pawlowski
On Fri, 25 Apr 1997, Sandi Burany wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } } Hello Suppliers: } } Would you please send me the catalogs of the materials needed for TEM, } SEM experiments. } } Thank you in advance. } } Sandy Burany } CHIPWORKS } 3685 Richmond Rd, Suite 500 } Ottawa, ON K2H 5B7 } Canada } (613) 829-0414 Ext. 3056 } FAX: (613) 829-0515 } Email:sburany-at-chipworks.com }
Yves MANIETTE wrote: } I am interested in any hint in order to perform the calibration of a TEM } high temperature sample holder test, up to 1000 degrees C. The idea is } looking at various sample with a known structural transformation ocurring } at precise temperature, and compare this "real" temperature with the one } given by the sample holder thermocouple=8A
Be cautious=8A
We have often observed that phase transition in perovskites and memory alloys (in the range of 100=B0C to -200=B0C) did occur at significant lower temperature in TEM thin foils than in bulk material (some tens of =B0C). We do not believe that it is due to errors in the temperature calibration, nor temperature gradient in the sample/sample holder, or to a heating effect by the electron beam. The source of this effect has probably to beaccounted for in the reduced size of the sample (=892D phenomena) or the sample surfac= e quality (presence of an amorphous layer on top/bottom of ion milled samples for example).
Remember that 2.5nm gold particles suffer fron a "size effect" and melt below 300=B0C instead of more than 1000=B0C for the bulk!
Remember also that the beam may significantly increase the temperature of particles deposited on a film when they start to be absorbant (thick observable samples). We vaporized micrometers sized silver spheres deposited on a carbon film just by focusing the illumination beam (100 kV, W filament, largest standard condenser aperture)! But very small spheres were not significanltly heated because of their high surface/volume ratio more favourable to cooling by radiation than heating by absorption.
Good luck Philippe A. Buffat
__________________________________________________________________ Philippe Buffat Ecole Polytechnique Federale de Lausanne (EPFL) Centre Interdepartemental de Microscopie Electronique Address: EPFL-CIME, Batiment MX-C, CH-1015 Lausanne, Switzerland Phone: +41(21)693 29 83 Fax: +41(21)693 44 01 (Central European Time) E-mail: philippe.buffat-at-cime.uhd.epfl.ch, WWW URL http://cimewww.epfl.ch/ ______________________________ Eudora F2.1 ___________________________
I do regurlarly count of Retrovirus particles in negative staining for companies in the Biotech industry. Concerning count in thin sections, I tried, but I abdict because there were too much parameters giving false results: size of the pellet, type of grids, thickness of the sections, etc. Thin sections are good for know how many cells are infected and to determine clearly the type of Retrovirus particles. I have good results in negative staining. I made count in mixing supernatant containing Retrovirus particles with latex beads of known concentration. I ultracentrifuge the mix on a grid in a Beckman Airfuge at 20-30 psi for 5 minutes, and stain with PTA 3%, pH 6. This count in negative staining is more reliable and less expensive than thin sections. If you need more details, do not hesitate to contact me.
Robert Alain Institut Armand-Frappier Laval, Quebec tel: 514-687-5010 ext 4388
Robert_alain-at-IAF.uquebec.ca
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am interested in finding out some of the ways that people tend to quantify retroviral particles (as a whole, not just infectious). We have been using a vendor that performs thin-section TEM and have looked some into negative stain TEM. We have not seen consistent results from the thin section determinations, and have been working on the sample prep end to get a more consistent pellet volume and consistency to submit for TEM.
I am interested in hearing what other people tend to do for retroviral quantitation (particularly in the Biotech industry because of FDA concerns). If I could get some input from this listserv group, I would appreciate it. The address above is correct. Thank you.
new to the field of EM polymer blends I would like to ask you for the best way to prepare PE/PS blends for structure determination with TEM.
Of course the material has to be stained and cut, but:
- what is the best staining agent (OsO4, RuO4, or something else)? - is it better, first to stain and then to cut or the other way round? - which temperature is the best for cutting, liquid nitrogen or room temperature?
I would appreciate ever tip & trick you can give on handling this kind of material.
Tanks in advance
Petra
-------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public Centre Universitaire (CRP-CU) Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpcu.lu or 100112.2335-at-compuserve.com
Use our Web page, "LabNet" for a review of 89 products. it is http://www.shore.net/~catalogs
Regards, Elinor Solit Director of Publications The Microscope Book
On Fri, 25 Apr 1997, Sandi Burany wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } } Hello Suppliers: } } Would you please send me the catalogs of the materials needed for TEM, } SEM experiments. } } Thank you in advance. } } Sandy Burany } CHIPWORKS } 3685 Richmond Rd, Suite 500 } Ottawa, ON K2H 5B7 } Canada } (613) 829-0414 Ext. 3056 } FAX: (613) 829-0515 } Email:sburany-at-chipworks.com }
Dear All, I have a student doing Mossbauer and EM studies of commercial ferrosilicons. We have a need to check our microanalysis results with regard to Fe and Si in both the SEM and TEM. Does anyone have or know where we can get Fe2Si and Fe3Si (or if all else fails anything close) to make standards? Thanks Mike
Michael J Witcomb PhD Electron Microscope Unit University of the Witwatersrand Private Bag 3 WITS 2050 South Africa
We are contracting our topical issues for 1998, and have resolved all problems of the huge backlog that plagued us in 1995 and 1996. If you are interested in serving as a Guest Editor for a particular topic, please e-mail me direct and present your idea. There is an honorarium. All articles will be published within about 6 months after they are received by the publisher in New York, so hot research articles would fit in nicely.
John E. Johnson, Jr., Ph.D. Editor, Microscopy Research and Technique sdinfo-at-worldnet.att.net
NYMA Inc., an aerospace engineering company serving NASA at Lewis Research Center, is seeking to fill the following two challenging positions;
SCANNING ELECTRON MICROSCOPIST
Required to operate and maintain three scanning electron microscopes, and provide materials characterization support. The individual must also be able to assist and instruct research staff in the principles and operation of scanning electron microscopy.
Requirements: B.S. degree with 3 years experience or 7+ years of extensive hands on experience in electron microscopy. Must have a thorough working knowledge of x-ray energy/wavelength dispersive spectroscopy, preparation equipment, and vacuum systems. A working knowledge of x-ray diffraction and electronics is a plus. Excellent communication and interpersonal skills are required.
TRANSMISSION ELECTRON MICROSCOPIST
Extensive experience in operating, and maintaining a 200 KeV transmission electron microscope in support of material characterization of advanced high temperature materials. The successful candidate will work with materials researchers to understand materials' properties.
Requirements: Ph.D. in material science with 3+ years extensive experience operating a transmission electron microscope using SAED, CBED, XEDS, EELS, and PEELS to analyze a wide range of materials. The individual we seek must have extensive experience in sample preparation using electro-polishing, ion-milling, PIPS, and PIMS. A working knowledge of a dedicated STEM and x-ray diffraction is a plus. Excellent communication and interpersonal skills are required.
Qualified candidates should submit a resume to : Todd Leonhardt, NYMA, Inc., Mail Stop 105-1, 2001 Aerospace Parkway, Brook Park, OH 44142 or by E-mail to todd.a.leonhardt-at-lerc.nasa.gov
Posted Date: April 28,1997 Closing Date: Open until filled
With PE/PS I have gotten good results if I first cut and then stain. I do use cryo sectioning and I cool the sample to about -100C (room temperature will not work because the Tg of PE is too low). I use folding grids to collect my sections (dry) and then I stain with RuO4 which will stain the PS. You can also stain the block first and then cut but then you are limited to sections near the surface of the specimen (the stain does not penetrate too far). Even then, you might need to re-stain your sections. Finally, I deposit carbon.
I hope this helps,
Jordi Marti ----------------------------------
Hello polymer experts,
new to the field of EM polymer blends I would like to ask you for the best way to prepare PE/PS blends for structure determination with TEM.
Of course the material has to be stained and cut, but:
- what is the best staining agent (OsO4, RuO4, or something else)? - is it better, first to stain and then to cut or the other way round? - which temperature is the best for cutting, liquid nitrogen or room temperature?
I would appreciate ever tip & trick you can give on handling this kind of material.
Tanks in advance
Petra
-------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public Centre Universitaire (CRP-CU) Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpcu.lu or 100112.2335-at-compuserve.com
I have two questions about type of oil you use for mechanical and diffusion pumps for the microscopes.
(I) We have a JEOL SM 35-C (SEM) since many years ago. In the last two years we have often seen oil drops condensed on the EDS detector, inside the chamber (at least it looks like oil, light green-dark yellow coloured drops). We've been using Dow Corning 704 silicon oil for the diffusion pump since ever. Now, we're not sure what it is going on, we suspect either: * backstreaming problems from the mechanical pumps oil * not sufficiently high vapor pressure for the DC 704
What do you think about it? any experience?
(II) We have been using Edwards 15 oil for the mechanical pumps. This time we have gotten a very high quotation (according to our reduced budget) for this oil. We're thinking of looking for other brand of the same type to replace it. Any idea what to buy?
We would appreciate to hear any suggestion from any of you.
Many thanks in advance. Sincerely,
Silvia Montoro Centro Regional de Investigacion y Desarrollo Guemes 3450 Santa Fe - Argentina csedax-at-arcride.edu.ar
NYMA Inc., an aerospace engineering company serving NASA at Lewis Research Center, is seeking to fill the following two challenging positions;
SCANNING ELECTRON MICROSCOPIST
Required to operate and maintain three scanning electron microscopes, and provide materials characterization support. The individual must also be able to assist and instruct research staff in the principles and operation of scanning electron microscopy.
Requirements: B.S. degree with 3 years experience or 7+ years of extensive hands on experience in electron microscopy. Must have a thorough working knowledge of x-ray energy/wavelength dispersive spectroscopy, preparation equipment, and vacuum systems. A working knowledge of x-ray diffraction and electronics is a plus. Excellent communication and interpersonal skills are required.
TRANSMISSION ELECTRON MICROSCOPIST
Extensive experience in operating, and maintaining a 200 KeV transmission electron microscope in support of material characterization of advanced high temperature materials. The successful candidate will work with materials researchers to understand materials' properties.
Requirements: Ph.D. in material science with 3+ years extensive experience operating a transmission electron microscope using SAED, CBED, XEDS, EELS, and PEELS to analyze a wide range of materials. The individual we seek must have extensive experience in sample preparation using electro-polishing, ion-milling, PIPS, and PIMS. A working knowledge of a dedicated STEM and x-ray diffraction is a plus. Excellent communication and interpersonal skills are required.
Qualified candidates should submit a resume to : Todd Leonhardt, NYMA, Inc., Mail Stop 105-1, 2001 Aerospace Parkway, Brook Park, OH 44142 or by E-mail to todd.a.leonhardt-at-lerc.nasa.gov
Posted Date: April 28,1997 Closing Date: Open until filled
I am interested in studying silver deposited onto Pt(111) by STM in electrochemical conditions.Does anybody tried (and succed) to obtain EC-STM images with this system ? I didn't find any publication on this subject. I know that there are allready publications with this system studied in UHV but my results are far away from what they published.
Thank you very much for your cooperation. ------------------------------------------ :-) :-) :-) :-) :-) :-) :-) :-) :-) :-) :-) :-) ------------------------------------------ Lorena H. Klein Laboratoire de Physico-Chimie des Surfaces (CNRS-URA 425) 1, Pl. A. Briand F-92195 Meudon FRANCE tel: +33 1 45075361, fax: +33 1 45075858 e-mail: lorena.klein-at-cnrs-bellevue.fr
I have a good reference that you might want to check out:
"Ruthenium Tetraoxide Staining of Polymers for Electron Microscopy", by J.S. Trent, et al, Macromolecules Vol. 16, #4, pp. 589- 1983.
This article is VERY informative.
Regards,
Bob *********************************** Bob Citron Chiron Vision 555 W. Arrow Hwy Claremont, CA 91711 ph: (909)399-1311 email: Bob_Citron-at-cc.chiron.com ***********************************
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello polymer experts,
new to the field of EM polymer blends I would like to ask you for the best way to prepare PE/PS blends for structure determination with TEM.
Of course the material has to be stained and cut, but:
- what is the best staining agent (OsO4, RuO4, or something else)? - is it better, first to stain and then to cut or the other way round? - which temperature is the best for cutting, liquid nitrogen or room temperature?
I would appreciate ever tip & trick you can give on handling this kind of material.
Tanks in advance
Petra
-------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public Centre Universitaire (CRP-CU) Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpcu.lu or 100112.2335-at-compuserve.com
Message-Id: {1.5.4.32.19970428230321.0068b974-at-pop3.unsw.edu.au} X-Sender: s7001031-at-pop3.unsw.edu.au X-Mailer: Windows Eudora Light Version 1.5.4 (32) Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
} -------------------------------------------------------------- } Dr. Petra Wahlbring } Centre de Recherche Public Centre Universitaire (CRP-CU) } Laboratoire d'Analyse des Materiaux (LAM) } 162a, av. de la Faiencerie L-1511 Luxembourg } tel. +352-466644-402 fax +352-466644-400 } e-mail: petra.wahlbring-at-crpcu.lu or 100112.2335-at-compuserve.com } } Dear Petra,
The book you must read is POLYMER MICROSCOPY by Linda Sawyer and David Grubb Chapman & Hall 2nd edition 1996 ISBN 0 412 60490 6
Try Chapman & Hall 2-6 Boundary Row, London SE1 8HN UK
one of the problems associated with lead citrate stain preparation is the presence of carbonate in the NaOH. John Luft came up with a solution for this years ago. Here is a copy of a handout from his 1976 seminar series that covers both uranyl acetate and lead citrate stain preparation.
steve
{bold} {fontfamily} {param} Times {/param} {bigger} {bigger}
NOTES ON URANYL AND LEAD STAINING (J.H. Luft) {/bigger} {/bigger} {/fontfamily} {/bold} {fontfamily} {param} Times {/param} {= bigger} {bigger}
{bold} {underline} Uranyl Acetate
{/underline} {/bold}
A saturated solution of uranyl acetate in distilled water is commonly used to stain EM sections either alone or preceding a lead stain.=20 However, uranyl acetate is not a well-behaved salt either as crystals in the bottle or as a stock solution. It tends to hydrolyse to form the basic acetate, U02(OAc) 2.2H20 + H20 --} U02(0H) (0Ac) + H0Ac and free acetic acid which produces the smell of acetic acid in a bottle of crystals. The easiest way to detect the presence of the basic acetate is to dissolve some of the uranyl acetate in question; at room temperature, it should dissolve (overnight) to give a crystal clear 8% solution. Any pale yellow residue on the bottom is the basic acetate, and the solution made up with this uranyl acetate will be saturated with the basic acetate. This basic acetate probably is the main source of contamination of EM sections from uranyl staining. Even originally completely clear uranyl acetate solutions may begin to decompose to precipitate out the basic acetate, but it is erratic - a week, or a year.
Large crystals of uranyl acetate seem to be much more stable against hydrolysis form moisture in the air than broken crystals. Powdered uranyl acetate is the worst offender in this regard, although {underline} fresh {/underline} powdered material may be satisfactory. In the past, uranyl acetate from Fisher Chemical Company has been the best because it is supplied as relatively large crystals. If no source of easily soluble uranyl acetate can be obtained, it is possible to recrystallize good material from even a badly hydrolysed supply as follows:
About 50 gm of the decomposed uranyl acetate is added to 100 ml of 10% acetic acid and heated to 70-80 {/bigger} {/bigger} {/fontfamily} {bigger} {bigger} {fontfamily} {param} MS_Li= neDraw {/param} =DF {/fontfamily} {fontfamily} {param} Times {/param} C on a hot plate with stirring for 10-15 minutes. the hot solution is then filtered through a conical paper filter into a clean, covered beaker and allowed to cool with frequent stirring, otherwise the crystals form on the sides and bottom of the beaker). It can be cooled further with ice for a higher yield. After several hours, the crystals can be filtered and sucked dry by vacuum for an hour on a Buchner funnel and stored in a tightly capped bottle. The residue from the original 50 gms. can be added to more old uranyl acetate and another batch recrystallized. This recrystallized uranyl acetate should give clear solutions for several years.
{bold} {underline} Lead Stains
{/underline} {/bold}
Reynolds alkaline lead citrate (E.S. Reynolds, J. Cell. Biol. {underline} 17 {/underline} , 208, 1963) is one of the most successful of the alkaline lead stains because of its intensity and relative freedom from contamination. Nevertheless, problems do arise with it, and contamination may appear mysteriously. Most of the trouble, I believe, is due to the problems of storing strongly alkaline solutions. These=20 rapidly attack and dissolve silicate from glass bottles (including Pyrex), and pick up C02 from the air if they are stored in polyethylene bottles. Solutions of Reynolds stain stored in small polyethylene bottles deteriorate and develop crystals or a residue on the side of the bottle in several months. It can be shown that this is a result of the C02 absorption, because a similar polyethylene bottle of stain kept {underline} within {/underline} a larger, wide-mouth, bakelite-capped bottle, which contains a little moist barium hydroxide as a C02-trap, remains clear and useful for several years.
The crystals are not a direct result of carbonate accumulation itself, because sodium carbonate can be dissolved directly in Reynolds stain without precipitation. Instead, the absorbed C02 acting as an acid probably reduces the pH of the stain to the point that the lead complex precipitates, and where the stain then does become sensitive to carbonate precipitation.
The nesting of a polyethylene bottle inside a glass bottle combines the alkali-resistance of polyethylene with the C02-impermeability of glass. This is an ideal solution for storage of alkaline materials but the solutions should be C02-free when they are made up as well. Reynolds advises the use of freshly boiled distilled water for this reason, and this works well. (Many stills can actually {underline} concentrate {/underline} C02 in the distillage, and the C02-content of some tap water varies from winter to summer, so boiling is a useful habit). It is satisfactory to boil a liter of distilled water in an Erlenmeyer flask down to 600-700 ml, then cover with a beaker and cool in tap water and use within a few hours.
Boiling water and waiting each time to make Reynolds stain is a nuisance, and it would be useful to make up stock solutions once with boiled distilled water and have it on hand thereafter. This is the recipe:
{bold} Solution A {/bold} : Lead nitrate, Pb(N03)29 31.67 gm made up to 500 ml with boiled, distilled water, plus 10 drops concentrated nitric acid.
{bold} Solution B: {/bold} Sodium citrate, Na3 C6H507.2H209 41.9 gm made up to 500 ml with boiled, distilled water. Add 5 drops of {underline} solution A {/underline} as a preservative. Both can be stored in regular capped glass bottles. The stain is made by adding 21 ml of {underline} solution A {/underline} to the 50 ml clean polyethylene bottle, using a 25 ml graduate. Rinse the graduate with distilled water and then add 21 ml of {underline} solution B {/underline} , and swirl to mix. Add 8 ml of 1.0N Na0H and again swirl. The solution clears and produces 50 ml of Reynolds lead stain ready for use.
The source of the 1.0N Na0H is also a problem, since C02 adds rapidly to pellets of Na0H or to a stock solution. (10 N Na0H is available "carbonate-free" in polyethylene bottles. It may have been carbonate-free when it was made, but there is plenty of carbonate in it when it arrives). A neat device for ready access to carbonate-free Na0H at any time is the following:
Take a clean 250 ml polyethylene bottle and fill it about half full of Na0H pellets (from a freshly opened bottle, but this is not essential). Then fill it about 3/4 full with distilled water. Quickly cap and submerge in cold running tap water because much heat is generated.=20 Shake for an hour or two and then allow to stand on the laboratory shelf for two weeks or longer. This produces saturated Na0H and a layer of transparent Na0H crystals will develop between the pellets below and the solution above. With a {underline} plastic {/underline} 20 ml syringe and large (15 ga.) needle, suck up 13.0 ml of the saturated solution and dilute to 250 ml with boiled distilled water. Store this in a polyethylene bottle enclosed in a larger wide-mouth jar capped tightly and containing moist barium hydroxide. This combination keeps the NaOH carbonate-free indefinitely. Be sure to rinse the plastic syringe several times to remove the silicone oil lubricant, or use a de-greased plastic syringe. The saturated NaOH rapidly attacks a glass syringe. This works because although the polyethylene bottle is porous to C02, the saturated Na0H precipitates out the Na2C03 which continuously collects but floats on the surface. Therefore, don't shake the saturated Na0H before using it.
A question for the Microscopy Listserver on behalf of David Grattan;
--------------------------------------------------------------- We are dealing with an antigen in brain tissue that seems to be adversely affected by paraformaldehyde, and would like to find a gentler fixative that still preserves neuronal morphology.
Has anybody tried fixation with carboiimide (1-ethyl-3-(3-dimethylaminopropyl)carodiimide). We seem to recall a reference to this as an EM fixative that also provided good morphology for immunohistochemistry, especially for neurons.
Any suggestions/ideas regarding this method of fixation would be appreciated. ------------------------------------------------------------------ Dr. David Grattan Department of Anatomy and Structural Biology School of Medical Sciences, University of Otago P.O. Box 913, Dunedin, New Zealand Ph: (64)(3) 479-7442 (office), 479-7345 (lab) Fax: (64)(3)479-7254 Email: david.grattan-at-stonebow.otago.ac.nz Homepage: http://www.otago.ac.nz/ResearchPostGrad/Neuro/Grattan _______________________________________________________________
----------------------------------------------------------------------- Richard Lander Senior Technician South Campus Electron Microscope Unit c/- Pathology Department Otago Medical School P.O. Box 913 Dunedin New Zealand. Tel. National 03 479 7301 Fax. National 03 479 7254
"Southernmost EM Unit in the World!" ------------------------------------------------------------------------
Message on behalf of Richard Easingwood; } } We are looking at the various options for large-format (4 x 5 inch) } negative scanners and wonder whether we could get some feedback from actual } users. } I have heard that the Leaf 45 scanner (which I understand was the Rolls } Royce/Cadillac of scanners) is discontinued and no longer available - can } anyone confirm this? } } Does anyone have experiences (good or bad) with the Polaroid SprintScan 45? } } We want to use the scanner for getting high quality resolution scans from } our sheet film TEM micrographs. } } Thanks in advance for any feedback. } } Regards, } } Richard } } Richard Easingwood } South Campus Electron Microscope Unit } School of Medical Sciences } University of Otago } PO Box 913 } Dunedin } NEW ZEALAND } } Telephone: 64-03-479 7301 } Facsimile: 64-03-479 7254 } } e-mail: richard.easingwood-at-stonebow.otago.ac.nz } } }
I'm sorry about the inconvenience caused by my unreadable mail. Thanks to our friendly neighbourhood SysOp I think I have found the problem (if You can read this). My postal adress somehow ended up in the field for the reply adress. If You still have an undeletable Email somewhere You can get rid of it by editing the mail file in a word processor. -- Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 86 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www_scem.csb.ki.se/pages/philip.html
I have just read the interesting post of Steve Barlow in which he gives John Luft's method for re-crystallizing Uranyl acetate to minimize stain artefact. The logic seems pretty good and I am willing to accept it. I am not all that eager, however, to run out and waste a couple hours recrytallizing a relatively toxic powder. This seems to me the perfect thing for an EM supplier to offer. They all sell UA but wouldn't it be nice if someone sold stocks of UA with certified recrystallization dates? I would be willing to pay a little extra for that. Furthermore, why doesn't any supplier sell UA in smaller aliquots in rubber stopper vials so that one could make up small amounts without having to worry about students spilling toxic dust all over the scale, etc. I know a lot of EM suppliers monitor this list and would be pleased to see them responsed.
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
} } We are looking at the various options for large-format (4 x 5 inch) } } negative scanners and wonder whether we could get some feedback from actual } } users. } } I have heard that the Leaf 45 scanner (which I understand was the Rolls } } Royce/Cadillac of scanners) is discontinued and no longer available - can } } anyone confirm this? } } } } Does anyone have experiences (good or bad) with the Polaroid SprintScan 45? } } } } We want to use the scanner for getting high quality resolution scans from } } our sheet film TEM micrographs. } } } } Thanks in advance for any feedback. } } } } Regards, } } } } Richard } } } } Richard Easingwood } } South Campus Electron Microscope Unit } } School of Medical Sciences } } University of Otago } } PO Box 913 } } Dunedin } } NEW ZEALAND } } } } Telephone: 64-03-479 7301 } } Facsimile: 64-03-479 7254 } } } } e-mail: richard.easingwood-at-stonebow.otago.ac.nz } } } } } Recently there have been several postings about negative scanners. We have been using the Agfa Arcus II for the last two years with somehat satisfactory results, however, we cannot seem to scan lattice images when they are taken below 340kx. Agfa now has a new product, the Duoscan that may allow one to do this still at a reasonable price. The Arcus II is a 12 bit scanner with 600x1200 dpi optical resolution and currently sells for about $1,800. The Duoscan has a better resolution I think around 2000x2000 dpi and I believe sells for about $5,000. We are trying to evaluate it right now.
Thanks so much to all the colleagues who took the time to offer references, web sites, comments and answers to the questions for Kelly's intergrated science project. She in compiling data, reading material, visiting web sites and getting other suggested sources. The wonderful thing is that she is learning and is so enthusiastic about the scientists all over the world who know EM and were kind enough to point her in the right direction. From the wilds of Africa to the shores of France and Australia and right here in the U.S. (and many other countries), there is a bond that has formed via science, and Kelly will always remember that people cared. What wonderful mentors you have been!!
Thank you all sincerely, Linda M. Fox lfox1-at-wpo.it.luc.edu
At 04:46 PM 4/29/97 +1200, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Yes, this is true. However, you may be able to locate a refurbished Leaf 45 Scanner by contacting a local digital photography distributor.
} } } } Does anyone have experiences (good or bad) with the Polaroid SprintScan 45? } } } } We want to use the scanner for getting high quality resolution scans from } } our sheet film TEM micrographs. } } } } Thanks in advance f snip, snip
A new option which I have not seen discussed on this forum is a drum scanner. The company ScanView which has made high-quality drum scanners for the pre-press market has now begun to market cheaper drum scanners at a relatively low price. We recently purchased the Scanview 3000 (3000 dpi, ~$15,000) which has an optical density range of 3.6, color resolution of 3 x 12 bit, 4096 levels, and a scanning area of 8.5" x 11.5". Some advantages of this scanner over the Leaf 45 are that the 3000 dpi resolution is available over the entire drum and the scan speed is much faster at the same resolution since it is a single pass scan. So, enlargements of an area of the negative which is off center are easy to do. You can also mount up to 6 negatives at once and set up batch jobs which will free up some of your time. We have been very pleased with the results so far. I believe it will deliver all possible information from the negative since the resolution of negatives are not much better than 3000 dpi in most circumstances. ScanView also makes two cheaper drum scanners the Scanmate Plus II (2600 dpi, o.d.=3.6) and the Scanmate Magic (2000 dpi, o.d.=3.0). I suggest that you contact your local digital photography dealer for a demonstration.
I have no financial interest in ScanView. I am just a satisfied user.
******************************************************* David F. Teter Los Alamos National Laboratory Materials Science and Technology: Metallurgy (MST-6) Mail Stop: G755 Los Alamos, NM 87545 ph: (505)665-0160 fax: (505) 665-0657 e-mail: teter-at-lanl.gov *******************************************************
I would like to ask several questions and I would be pleased if someone can answer one of them.
I want to capture images with an infrared film and capture only emitted wavelengths.
1) What literature can I read something about infrared emission wavelengths ?
2) What filters are available to select only a certain range of infrared wavelengths ?
3) Does anybody know any application of infrared photography in scientific work ?
Thank you for your attention.
Rui
/-------------------------------------------------------------------------\ | Rui Costa | e-mail: ruicosta-at-esb.ucp.pt | | Escola Superior de Biotecnologia |telephone: 351-2-5580044 | | Rua Dr Antonio Bernardino de Almeida |fax: 351-2-590351 | | 4200 PORTO | | | PORTUGAL | | \-------------------------------------------------------------------------/
Here's a can of worms if there ever was one! I'll infer that the reasons for the inquiry are that a) someone, probably your boss, has told you that you are not doing enough and b) you are already working as hard/fast as you can. Rate (call it turn around time or whatever) is only ONE measure of productivity. If it is the ONLY measure of productivity then your manager is failing to do his/her job properly. Turnaround time has to be evaluated along with quality, continuous improvement, customer satisfaction, profit/loss, employee morale, customer value, and a host of other criteria that are all inter-related. The analysis does not occur in a vacuum (bad pun) isolated from the rest of the world. You and your work are part of a system. Changing ANY part of that system WILL disrupt another part.
There has to be a consensus among ALL stakeholders as to what is an acceptable "workload." This consensus must be reached using FACT, not speculation, which can be obtained by benchmarking your lab against equivalent labs, then comparing best/worst cases. Of course the irony is that you be taking time away from the scope to look at best practices, etc. By including all stakeholders in a rational, calculated, cross-functional, balanced approach, everyone will have a better understanding of what can/can't be done and why/why not.
I know this answer does not give you a number and that is what you were looking for, but without a balanced approach including all stakeholders using hard data as well as personal input, you be in an increasingly tense loop leading to major problems.
Read anything by W. Edwards Deming, Peter Senge, Chris Argyris, Peter Drucker, Margaret Wheatley (sp?), Philip Crosby, Tom Peters....
IF YOU SPECIALIZE IN IMMUNOCYTOCHEMISTRY/IMMUNOHISTOCHEMISTRY/OTHER RELATED AFFININITY METHODS, may I URGENTLY draw your attention to my PROPOSED NEW NEWSGROUP to be called "sci.bio.immunocytochem"
The formal proposal can be found posted in the USENET NEWSGROUPS "news.announce.newgroups" and "news.groups". The latter contains material of a rather varied nature (!) but it is necessary to use this unmoderated group to hold set-up discussions for proposed new groups.
If you are keen to see A NEW IMMUNOCYTOCHEMISTRY NEWSGROUP, then please go to "news.groups", ignore all the rubbish, look carefully for the articles with "sci.bio.immunocytochem" in their subject line. By the time you read this message, the CALL FOR VOTES may well be underway. When you see "CFV: sci.bio.immunocytochem" (but not before please) then it is time to VOTE! You won't need access to Usenet to vote for my new group, only e-mail. We need at least 100 VOTES to get the group up and running!
For those people who are unfamiliar with Usenet, Newsgroups are a bit different from the Web or E-Mail. It is necessary to have access to "Usenet" which not all academic computer departments offer. So first check it you have access. Next you will need "news-reading software" which is available free by downloading it from the Web (or ask your computer department if they have some on a floppy disc). Once you have your "newsreader" (eg Free Agent for Windows) then you can read and post to newsgroups easily.
It is possible to access news via the Web, for example with Deja-news, but it is realy clunky and a last resort in my opinion. However, you CAN read the official proposal at the Royal Microscope Society web site {http://www.rms.org.uk} , or at the Introduction to Immunocytochemistry (Center for Cell Imaging Department of Cell Biology Yale University School of Medicine) web site {http://info.med.yale.edu/cellimg/CCIimmuno.html}
Proponent: Amanda Wilson {awilson-at-aw.u-net.com}
I am working with distarch phosphates (carbohydrate chains covalently linked together by Phosphorous), created with POCl3. I was hoping that someone can tell me whether it would be possible to use some type of probe (maybe with fluorescence?) to be able to detect the phosphorous based crosslinks. I have used SEM with EDS and this technique is not sensitive enough to detect the low level of phosphorous in the starch.
The oil drops on your EDS detector could be from either the diffusion pump, or from the rotary vane backing pump, or from both, and I don't know any easy way to tell which. If you can collect a few drops of it you might try putting it on a specimen stub and running an EDS analysis on it. If you find silicon to be present then you know that at least some of it comes from the diffusion pump. Even better, perhaps you can find someone in some chemistry lab that will run an infra red spectrem of it for you. That should give a difinitive identification.
In the past, we have had trouble with oil collecting on the EDS detector in one of our SEMs. and an FTIR spectral analysis showed it to be mainly from the rotary vane backing pump. Ed Birko, of the Hitachi service organization, has told me that on several occasions he has also found that the roughing pump was the major source of such contamination. As discussed on p. 144 of my book, "Vacuum Methods in Electron Microscopy," the molecules of the roughing pump oil get broken down into volatile low molecular weight fragments by the mechanical rubbing of the parts inside the pump, and these fragments readily migrate into the backing line and thence through the diffusion pump and into the vacuum system. Most diffusion pump oils, and particularly the silicone oils, are quite resistant to thermal and oxidative degradation, and so are unlikely to be a major contributor unless the vacuum system on your instrument is poorly designed or your diffusion pump has been subjected to one of the improper operating situations described on p. 214-223 of VM in EM.
There are two principal ways of dealing with contamination problems arising from the roughing pump. You can give your instrument a good cleaning and then install a good foreline trap, which must then be faithfully maintained to preserve its effectiveness (VM/EM p.147-149). [One approach that appears reasonably effective is to use the Micromaze traps sold by the Lesker Co,, heating them continuously with the heaters normally used for regeneration running at about 75% full power (VM/EM p. 147)] Alternatively, you can use an inert gas purging system, which is brought into operation when the instrument can be put into the standby mode, to attempt to sweep the oil molecules back out of the vacuum system - VM/EM p. 145 (XEI sell a system designed to perform this operation automatically).
In any event, you will need to remove the presently contaminating oil from your EDS detector window, otherwise it will begin to alter your detector sensitivity. This can be a very tricky process, and should be done with a great deal of care. It is best to check with the manufacturer of your detector for their recommendations before you try anything. (We have successfully removed oil from detectors with the standard beryllium windows by VERY CAREFULLY running a few drops of petroleum ether down over the window. Do NOT use a solvent such as acetone, or an alcohol, which may attack the epoxy normally used to hold the window in place, and be very careful not to puncture the window.)
Good luck,
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
I agree with Tom Phillips-perhaps our suppliers could add some comments for us about age of these materials and possible smaller quantities available. It's been a long standing problem. Grace (Hello Stacie, Ted, Steve.....?)
Dear Folks, Please accept this message in hopes that it will answer most of your questions posted to me in the past two weeks. The real problem with Pb staining for epoxy sections is that almost nothing is known about the mechanism of heavy metal deposition on epoxides for TEM. We must first consider the interaction between the very active monomers in the section with the Pbcit. About 10% of all monomers stay unbound during a typical embedment. These monomers react with a variety of heavy metals - I have never had the time to prove that they will react detrimentally(for TEM) with Pbcit. However, I have repeatedly witnessed the situation where lead stain which did fine with random sections, repeatedly, consistently "dumped" when used on severely underpolymerized sections. Depending on the typed of tissue embedded and the care with which it was processed, much more than 10% of monomers remain unpolymerized. (This can be a huge problem when staining thick sections with the methylene or toluidine blues, since the monomers will engage the chromogens in undesirable redox shifts, sometimes resulting in rapid fading). So - before we even get to the lead, we have a vacillating situation. We must also note that not all Shell Epon 812 substitues are true substitues. Some replacements contain dilutents. This will affect the final staining nature of the sections, perhaps positively, perhaps negatively. We just do not know.
The pH of the lead solution is critical. Over four years of accurate record keeping, I was able to gradually intensify the Pb stain by dropping the pH. Finally I had nothing but precipitate. By increasing the pH of the lead stain, I could eventually eliminate all staining activity from the control sections.
One must control the pH. How this is done is immaterial. Very close control may be difficult when pellets are used - they are difficult to weigh, they absorb water. If one can use pellets and then titrate to the same pH every time, one will get reliably good results. If the relationship with pH and lead citrate is not understood, and pellets are used, a laboratory may experience variations in Pbcit staining intensity. (In 1983 I received over 400 requests for reprints for a simple paper I wrote regarding the use use of reliable staining practice. Over the years I have suggested that laboratories who turned to me with their propblems to do two things immediately: 1. Abondon pellets in favor or commercially titrated NaOH 2. Chealate the citrate and lead adequately by shaking and inversion for at least 25 minutes. This has solved most problems immediately.) If one uses commercially titrated NaOH, the worry about pH variations disappears.
It is thought that stored NaOH absorbs carbon dioxide from the atmosphere resulting in lead carbonate precipitate. Probably happens. However, I simply have not had any problems with it. Moreover, once when I accidentally bubbled about 10ml of air through about 5ml of Pbcit solution in a staining dish containing valuable sections, I expected the worst. Nothing happened. We breathe on our sections sitting in lead drops, etc. We do not boil water. It may be possible that if the Pb is at the correct pH and also the Pb is well chealated to the citrate (with adequate shaking and inversion),this problem is minimized. We do not worry about carbonate.
Well chealated lead citrate does not keep well in glass containers. We keep our lead solution in syringes in the refrigerator for 6 months. At this time I am using lead stain which I made up 18 months ago. I am still using it successfully. I do not recommed this, however. I am just very curious to how long it takes for the system to fall apart.
The original paper by Reynolds is wonderful storehouse of information. I recommend to everyone who is having any problem to get it and study it. Tomorrow I am sending out the chapter to those who requested it. If you did not get it in about 10 days (allow for snail mail), let me know.
If someone has unsolvable problems, please feel free to call me. I might be able to pinpoint something which is not obvious, since I (and my students) have made about every mistake possible with this lead soup. My phone # is 303-871-3026.
Bye. See you all in Cleveland at the MSA meeting, I hope! Hildy
H. Crossman wrote: } } Here's a can of worms if there ever was one! I'll infer that the } reasons for the inquiry are that a) someone, probably your boss, has } told you that you are not doing enough and b) you are already working as } hard/fast as you can. Rate (call it turn around time or whatever) is } only ONE measure of productivity. If it is the ONLY measure of } productivity then your manager is failing to do his/her job properly. } Turnaround time has to be evaluated along with quality, continuous } improvement, customer satisfaction, profit/loss, employee morale, } customer value, and a host of other criteria that are all inter-related. } The analysis does not occur in a vacuum (bad pun) isolated from the } rest of the world. You and your work are part of a system. Changing } ANY part of that system WILL disrupt another part. ...
I agree! Of course all of this depends on the environment you are in and what is expected from the results. One reason I do TEM in a research environment is that I am better at quality than quantity.
A few years ago when I was still very new to EM, I learned my lesson. I had started feeling bad about being a tortoise instead of a hare. I kept reading about all the quick processing and embedding techniques that are used in clinical labs, and thought "wow, I could save days of work!". Wrong. All I did was waste a lot of time and mess up a few months' worth of samples. My necessarily large blocks of tendon and ligament just didn't behave like small biopsy specimens. It turned out when I skimped on time, I got poor fixation, infiltration, staining, etc. and ended up with too many artifacts to do the image analysis necessary for my research project. I had to repeat many of the samples. So I don't mind being a tortoise any more; it might just save time in the long run. (Then there are the additional benefits of hiding in one's shell - read darkroom - when the boss comes by! If I could just get over my fear of can openers ... but that's another matter ...)
-Karen
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Life Sciences Sector Lab Reply: kszaruba-at-mmm.com 3M Company 3M Center 270-1S-01 Phone: 612-737-2971 St. Paul, MN 55144-1000 Fax: 736-1519
These opinions are my own and may not represent those of 3M.
We have a NORAN (Tracor Northern) 5400 Series II EDS system (Model Number TN-5402/BBAA) acquired in 1987. It does not have a hard drive and operates off of two floppy drives. It requires 5.25" double sided, high density floppy disks, the originals of which are no longer readable. They have not been in use other than to be the source of copies made periodically but apparently they have simply deteriorated with time. We have copies of copies that are working pretty well but I'm becoming concerned that they may crash/die and we'll have no good source from which to make new copies. NORAN does not have replacement masters of this vintage so I'm hoping to find someone who has original master disks in good working order who would be willing to loan them to be copied.
We need two disks of the set from the November, 1987 Release 1C. What will work with our system is very specific and the original label on the master disks reads:
SERIES II SOFTWARE RELEASE 1C NOV., 1987, TRACOR NORTHERN, INC.
Of the disks in this release, we need:
SQ/SSQ/PRZ/ZAF Master NOT BOOTABLE
and
MSCAN2 Master
Thanks in advance for any help or suggestions you may have.
___________________________________________________________________________ Barbara Reine, Botany Dept. Box 351330 Univ. of Washington, Seattle, WA 98195-1330 e-mail: reine-at-u.washington.edu; ph: (206) 543-1955 ____________________________________________________________________________
Does anyone out there know the origins of the word "Wehnelt". I have seen it stated in print that it is named after its inventor, though this individual was not identified. I have also seen it stated that it is simply the German word for "grid". I can confirm neither of these statements. Does anyone out there know for sure? If it was named after the inventor, who was this person? -- Fred Schamber ....................... mailto:fhscham-at-SGI.NET
I am trying to find a source of plastic pages for three-ringed binders which have pockets large enough to hold polaroid prints (these are 131mm x 106mm). Ideally each page should have 4 pocket and be able to fit two prints back-to-back so images are visible from both sides of the page.
I would really appreciate it if someone could give me an address/phone/fax/email of a supplier of these items (if, indeed, these things are made). Cheers,
Mark Blackford TEM Group Materials Division, ANSTO PMB 1, Menai, N.S.W. Australia 2234 Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
} Does anyone out there know the origins of the word "Wehnelt". I have } seen it stated in print that it is named after its inventor, though this } individual was not identified. I have also seen it stated that it is } simply the German word for "grid".
My 2 cents on this is that "Wehnelt" most definitely does not mean "grid" in German. It has no uses in German whatsoever beside "Wehnelt-Anode" in electron microscopy. (The word for "grid" is "Raster", as in "Rasterelektronenmikroskop", which is abbreviated REM--hence the Germans' tendency to write REM when they mean SEM in English.) Although I don't know anything about the inventor, I always assumed that the Wehnelt anode was named after it's inventor. It sure sounds like it could be a German last name, albeit not a very common one.
We are about to start a program of work, using the TEM, on the respiratory tract of mammals/humans and I was wondering whether anybody out there could recommend any references on the same (especially for healthy but also for diseased tissue).
We already have copies of Rhodin, A.G. (Histology a text and atlas) but I was wondering if anyone knew of text books/atlases or review papers, on ultrastructure, which were more specific. Our general searches have produced little so I thought that the readership, with its combined experience of a small planet, may be able to help or suggest new areas to search.
If you think that your reply will not be of general interest, please reply to me directly by e-mail at the address below.
thanks
Malcolm Haswell University of Sunderland UK e-mail: es0mhs-at-environment.sunderland.ac.uk (NB. 3rd character is zero)
I bought my sleeves from a local camera supply house. Can your Polaroid representative help? The product I use is called:
NegaFile P.O. Box 78 Furlong, Pennsylvania, USA 18925
Reorder number (part number?) 4504
The sleeves hold four Polaroid prints. One caveat, make sure the prints are dry or they will stick to the plastic.
------------------------------------------------ Opinions or statements expressed herein, rational or otherwise, do not necessarily reflect those of my employer.
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 Phone: (508) 750-1717 E-mail: crossman-at-osi.sylvania.com
Our web sites: www.sylvania.com www.siemens.com --
I would like to know the experimental details of the gamma(electron) irradiation technique that has been used to enhance contrast in PE sections. Does anyone have this information ?
For the past two or three years, I have been using a text in my ultrastructure class entitled "Cell and Tissue Ultrastructure - A Functional Perspective," by P. C. Cross and K. L Mercer. It is a system by system atlas of ultrastructure. The chapter on the respiratory system has EM's on the mucosa, bronchioles, alveoli, type I and type II alveolar cells, components of the blood-air barrier, and alveolar macrophages. I have recommended the text to several of my colleagues throughout the state and they are now also using it. It has a 1993 copyright (ISBN 0-7167-7033-4) and is published by W. H. Freeman and Company. For the record, I have no interest, financial or otherwise, in W. H. Freeman or the authors of the text.
} We are about to start a program of work, using the TEM, on the respiratory } tract of mammals/humans and I was wondering whether anybody out there could } recommend any references on the same (especially for healthy but also for } diseased tissue). } } We already have copies of Rhodin, A.G. (Histology a text and atlas) but I } was wondering if anyone knew of text books/atlases or review papers, on } ultrastructure, which were more specific. Our general searches have } produced little so I thought that the readership, with its combined } experience of a small planet, may be able to help or suggest new areas to } search. } } If you think that your reply will not be of general interest, please reply } to me directly by e-mail at the address below. } } thanks } } Malcolm Haswell } University of Sunderland } UK } e-mail: es0mhs-at-environment.sunderland.ac.uk (NB. 3rd character is } zero)
Martin A. Levin Department of Biology Eastern Connecticut State University Willimantic, CT 06226 Phone: (860)465-4324 FAX: (860)465-5213
- ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear All,
I am trying to find a source of plastic pages for three-ringed binders which have pockets large enough to hold polaroid prints (these are 131mm x 106mm). Ideally each page should have 4 pocket and be able to fit two prints back-to-back so images are visible from both sides of the page.
I would really appreciate it if someone could give me an address/phone/fax/email of a supplier of these items (if, indeed, these things are made). Cheers,
Mark Blackford TEM Group Materials Division, ANSTO PMB 1, Menai, N.S.W. Australia 2234 Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
Mark,
We purchase polypropylene pages in serveral sizes to accomodate TEM negatives as well as SEM prints from 20th Century Plastics, P.O. Box 2376, Brea, CA 92622-2376, (800)767-0778. The following is a list of what we get: Item # EZV45-00 , EZ2C Archival holds 4 - 4x5" prints Item # EZVH4K-00, EZ2C Archival holds 6 - 4x6" prints Item # EZ116K-00, EZ2C Archival holds 12 - 3.5x4.5" negatives or prints
I hope this helps you. You can give them a call and they will send out a catalog of their products.
Pamela F. Lloyd Materials Science Engineer UES, Inc. 4401 Dayton-Xenia Rd. Dayton, OH 45432 (937) 255-1329 (Phone) e-mail: lloydpf-at-ml.wpafb.af.mil
Disclaimer: The views expressed in this e-mail message do not necessarily represent the official views of UES, Inc., and I have no connection to 20th Century Plastics other than being a user of their products.
Most of the original EM of lung (and most other organs) was done in the '60s and early '70s. Much of this material is too old to be in computer databases. Try the encyclopaedic Histology texts, Weiss for example, or Bloom and Fawcett. Also try the Handbook of Physiology; before embarking on the physiology of an organ they usually have a good chapter on LM and EM of it. Also try a big comprehensive textbook of pulmonary medicine, such texts often have an overview of LM and EM with references.
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (908)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
I may be dragging the thread away from contamination, but I thought that most electron microscope users avoided silicon based oils in diff pumps etc because they are extremely difficult to remove from the interior of an electron microscope and any contamination would normally have electrical insulating properties (catastrophic in an e.m.). Perhaps I am wrong but I would welcome any comments. After all this is one of the reasons why Santovac oils and their relatives became so popular (despite their costs).
If I am labouring under a mis-apprehension then I apologise.
Malcolm Haswell University of Sunderland UK ----------
The oil drops on your EDS detector could be from either the diffusion pump, or from the rotary vane backing pump, or from both, and I don't know any easy way to tell which. If you can collect a few drops of it you might try putting it on a specimen stub and running an EDS analysis on it. If you find silicon to be present then you know that at least some of it comes from the diffusion pump. Even better, perhaps you can find someone in some chemistry lab that will run an infra red spectrem of it for you. That should give a difinitive identification.
In the past, we have had trouble with oil collecting on the EDS detector in one of our SEMs. and an FTIR spectral analysis showed it to be mainly from the rotary vane backing pump. Ed Birko, of the Hitachi service organization, has told me that on several occasions he has also found that the roughing pump was the major source of such contamination. As discussed on p. 144 of my book, "Vacuum Methods in Electron Microscopy," the molecules of the roughing pump oil get broken down into volatile low molecular weight fragments by the mechanical rubbing of the parts inside the pump, and these fragments readily migrate into the backing line and thence through the diffusion pump and into the vacuum system. Most diffusion pump oils, and particularly the silicone oils, are quite resistant to thermal and oxidative degradation, and so are unlikely to be a major contributor unless the vacuum system on your instrument is poorly designed or your diffusion pump has been subjected to one of the improper operating situations described on p. 214-223 of VM in EM.
{snip}
Good luck,
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
Silvia & listreaders: In an oil pumped system some oil will always find its way into the chamber but you need to minimise this. Recent contributions dealt with rotary pump trapping. Condensation will be severe if your EDS has a pinhole in the Be. Then, while the SEM is at atmospheric pressure, the EDS snout becomes quite cold and when the SEM is pumped any oil will condense "extra well". Your vacuum system cooling may not be the best either. A liquid N2 trap above the dif pump would prevent the problem but that is not a simple solution. Is your cooling water flowing well (checked at the outflow) and what is its temperature? I would prefer the outlet water to be no higher than 20 degrees and if you have a recirculating system set the temperature to 14-16 degrees. Dif pump heaters are generally designed for those temperatures. The other bad news is your oil. Silicone is wonderful in coaters, unfortunately it react with the e- beam and produces totally insoluble products and they result in uncleanable apertures and other column problems. That is why the world switched to other fluids about 25 years ago. Most used is Santovac5 - even though these fluids are not as tolerant of abuse, (poor vacuum when pump is hot= no, no) nor are they as cheap as the silicones. If you do not know how to clean the Be window, send me an email. Best of luck. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 350+ Links, MSDS ************************ http://www.proscitech.com.au } } Hi everyone, } } } I have two questions about type of oil you use for mechanical } and diffusion pumps for the microscopes. } } (I) We have a JEOL SM 35-C (SEM) since many years ago. In the } last two years we have often seen oil drops condensed on the EDS detector, } inside the chamber (at least it looks like oil, light green-dark yellow } coloured drops). } We've been using Dow Corning 704 silicon oil for the } diffusion pump since ever. Now, we're not sure what it is going on, we suspect } either: } * backstreaming problems from the mechanical pumps oil } * not sufficiently high vapor pressure for the DC 704 } } What do you think about it? any experience? } } } (II) We have been using Edwards 15 oil for the mechanical pumps. } This time we have gotten a very high quotation (according to our reduced budget) } for this oil. We're thinking of looking for other brand of the same type to } replace it. } Any idea what to buy? } } } We would appreciate to hear any suggestion from any of you. } } } Many thanks in advance. Sincerely, } } } } Silvia Montoro } Centro Regional de Investigacion y Desarrollo } Guemes 3450 } Santa Fe - Argentina } csedax-at-arcride.edu.ar
Steven and listreaders: I would not dare to argue with the great John Luft - but sometime back in the times when Watson "invented" and Reynolds and others perfected lead stain, (and remember that UA only was good enough for methacrylates but not the then new epoxies) I shared my lead staining problems with an inorganic chemist. I learned that dilute NaOH solutions and NaOH pellets absorb CO2; which on pellets can show as a powdery white layer. Ever since I have used the top layer of pellets from a jar for other purposes and used fresh pellets in boiled (now I would sonicate) H2O. I also learned that 10N NaOH (which is 40%) is happy without CO2 and DOES NOT ABSORB it. I think that I only made three batches of 10N NaOH - after moving to another lab. Stored in a solid plastic bottle that stuff is good for "1000 years". A one plus nine solution just before use provides fresh, near CO2 free 1 N sod. hydroxide. I never measured pH of the final Reynolds solution. My lead preparations always worked contamination free until exhausted - about a year.
Similarly, the suggested elaborate process to recrystallise UA is baffling. For decades I used about 5ml of saturated UA in a dropper bottle with a single drop of HCl added and stored in the fridge for a maximum of one month. That preparation too was intrinsically trouble free.
All experienced biomed TEM workers have had Pb and UA problems from time to time but mostly, the reasons are simple failures of normal techniques. These failures can be perplexing at times but I do not believe that we need to re-invent Pb or UA methods. Regards Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 350+ Links, MSDS ************************ http://www.proscitech.com.au
one of the problems associated with lead citrate stain preparation is the presence of carbonate in the NaOH. John Luft came up with a solution for this years ago. Here is a copy of a handout from his 1976 seminar series that covers both uranyl acetate and lead citrate stain preparation.
......snip The source of the 1.0N Na0H is also a problem, since C02 adds rapidly to pellets of Na0H or to a stock solution. (10 N Na0H is available "carbonate-free" in polyethylene bottles. It may have been carbonate-free when it was made, but there is plenty of carbonate in it when it arrives). A neat device for ready access to carbonate-free Na0H at any time is the following:
Take a clean 250 ml polyethylene bottle and fill it about half full of Na0H pellets (from a freshly opened bottle, but this is not essential). Then fill it about 3/4 full with distilled water. Quickly cap and submerge in cold running tap water because much heat is generated. ......snip
I agree with Karen: a research environment is alot different than a clinical or quick-turn around lab. Research labs more often than not work with novel samples which require often novel approaches and slow, tedious processing as opposed to overnight processing of standardized tissue biopsies. Unfortunately, with major cutbacks for support of education and research, layers of accountants and administrators with no understanding of what goes on in these types of facilitities are having a larger say. I know it is our job to convince them that we cannot run identical to clinical or some commerical labs but it is getting extremely difficult to do so as when they only consider the bottom line ($$$). Of course this is not to say we shouldn't adopt ways to speed certain tasks in the lab, but these are usually in data base management and clerical duties. Getting back to the original question as far as work load, it depends on the lab type and particular study. I was once worked next to a histopath lab where 3 technicians cut paraffin blocks at about one per minute while another tech stained them (H&E) so the pathologists could read them by noon. This does not happen in a EM facility processing plant, fungal, spores, arthropods, embryos, assorted animal tissue, etc. for immunocytochemistry, autoradiography, image analysis, etc. and spending hours of beam time. Just my two cents Hank Adams EML New Mexico State University.
Hello to all, Many thanks to those who responded to my TEM stage drift inquiry.=20 Several of you requested a summary of the responses so here goes....there were ~10 responses from users & applications engineers.=20 The common denominator with respect to lack of stage stability was thermal stability as influenced by water temp. & air currents. =20 As to what levels of stability should be I expected from this instrument? The consensus is that within 10 - 30 min. the drift should be 1nm/min. or less & not more than a few nm/min. before that.=20 My compliments to those employed by vendors who replied. They were informative & I didn=92t suffer from salesman dispensing airware. =20
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Hi } } I would like to ask several questions and I would be pleased if someone } can answer one of them. } } I want to capture images with an infrared film and capture only emitted } wavelengths. } } 1) What literature can I read something about infrared emission } wavelengths ? } } 2) What filters are available to select only a certain range of infrared } wavelengths ? } } 3) Does anybody know any application of infrared photography in } scientific work ? } } Thank you for your attention. } } Rui } } /-------------------------------------------------------------------------\ } | Rui Costa | e-mail: ruicosta-at-esb.ucp.pt | } | Escola Superior de Biotecnologia |telephone: 351-2-5580044 | } | Rua Dr Antonio Bernardino de Almeida |fax: 351-2-590351 | } | 4200 PORTO | | } | PORTUGAL | | } \-------------------------------------------------------------------------/ } } } Hello Rui:
The easiest publications to start with are the Kodak series,N-17 "Infrared Films", M-28 "Applied Infrared Photography, N-1 "Medical Infrared Photography" and B-3 "Kodak filters for Scientific and technical use". You can get them through photography stores.
89B is the visibly opaque filter you would use to record just infrared. Since infrared film will record wavelengths in the visible spectrum as well as in the range of 700-900 nm.
We use infrared in dermatology to record the patterns of blood vessels in the skin. This is possible because the longer wavelengths can penetrate deeper.
Bob Underwood Morphology Core U of Washington Seattle
We buy our plastic storage pages from the University bookstore and also from local photography stores. They are archival quality and come in a variety of sizes. They are made by Print File Archival Preservers, PO Box 607638, Orlando Fl, Ph:407-886-3100; Fax 407-886-0008. The type I use for 4x5 polaroid prints is product #45-8P (which are a heavier weight than the negative preservers).
Hope this helps.
Rosemarie Rosell Assistant Research Scientist Dept. of Plant Sciences University of Arizona Tucson,AZ 85721 Ph: 520-621-1230 Fax: 520-621-8839
On Wed, 30 Apr 1997, Mark Blackford wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear All, } } I am trying to find a source of plastic pages for three-ringed binders } which have pockets large enough to hold polaroid prints (these are 131mm x } 106mm). Ideally each page should have 4 pocket and be able to fit two } prints back-to-back so images are visible from both sides of the page. } } I would really appreciate it if someone could give me an } address/phone/fax/email of a supplier of these items (if, indeed, these } things are made). Cheers, } } Mark Blackford } TEM Group } Materials Division, ANSTO } PMB 1, } Menai, N.S.W. } Australia } 2234 } Phone 61 2 9717 3027 } Fax 61 2 9543 7179 } } Disclaimer: } The views expressed in this E-mail message do not necessarily represent the } official views of ANSTO from which this message was conveyed. } } }
} I am trying to find a source of plastic pages for three-ringed binders } which have pockets large enough to hold polaroid prints (these are 131mm x } 106mm). Ideally each page should have 4 pocket and be able to fit two } prints back-to-back so images are visible from both sides of the page. } I would really appreciate it if someone could give me an } address/phone/fax/email of a supplier of these items (if, indeed, these } things are made).
These things are called "Polyview Pages" and are available sized to hold 4 Polaroids or 6 3-1/4" x 4" negatives. They are available from us, and, I believe, from several other EM supply companies.
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
What are some effective solvents for removing adhesive tape, spots, etc. from aluminum SEM mounts. Acetone is marginal as are other ketones. I've tried several alcohols without success. Caustics are out because they eat the aluminum. Pure and mixed acids don't work either. I've even tried WD-40 and liquid dish soap.
The ideal solvent would have low toxicity, low cost, etc... Water solubility would be a bonus. I'd really like a chlorinated solvent vapor degreaser! Fat chance.
Any suggestions? Terpenes, perhaps?
------------------------------------------------ Opinions or statements expressed herein, rational or otherwise, do not necessarily reflect those of my employer.
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 Phone: (508) 750-1717 E-mail: crossman-at-osi.sylvania.com
Our web sites: www.sylvania.com www.siemens.com --
YES, the do exist. We use a product called Kleer-Vu, Proline #14912 plastic, 3-ring binder pages for 4x5 prints. In fact, we use it for our Polaroid prints, back-to-back and it works beautifully. You should be able to purchase these at any professional graphic supplier. If you have problems finding a vendor, contact me again.
} I am trying to find a source of plastic pages for three-ringed binders } which have pockets large enough to hold polaroid prints (these are 131mm x } 106mm). Ideally each page should have 4 pocket and be able to fit two } prints back-to-back so images are visible from both sides of the page. } } I would really appreciate it if someone could give me an } address/phone/fax/email of a supplier of these items (if, indeed, these } things are made). Cheers, } } Mark Blackford } TEM Group } Materials Division, ANSTO } PMB 1, } Menai, N.S.W. } Australia
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 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 ####################################################################
} Does anyone out there know the origins of the word "Wehnelt". I have } seen it stated in print that it is named after its inventor, though this } individual was not identified. I have also seen it stated that it is } simply the German word for "grid". I can confirm neither of these } statements. Does anyone out there know for sure? If it was named after } the inventor, who was this person? } -- Dear Fred, About two months ago I replied to a Gustavo Waenelt about this. Here is my reply:
Harold: I have a product called Ease-Away, (manufactured by Wood Life Ltd, 9331 Park Lane, Franklin Park, IL 60131) which I bought from some mail order catalog a few years back whose advertising claims that it "removes adhesive and adhesive residue from any surface quickly, easily, and safely", and which I have, in fact, found to do a very good job on adhesive tape residues, but to work more slowly on removing tape itself (which is not surprising, since the tape represents a pretty heavy dose of plastic of some kind or the othger to dissolve).
Incidentally, I have also found that my pet cleaning agent, Tilex Soap Scum Remover, works pretty well on tape residues. As I've mentioned in previous comments on this listserver, it also does a pretty good job of removing silicone and polyphenyl ether diffusion pump oils from metal surfaces, paint from your hands, oil stains from carpets, food stains from clothing, etc.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
20th Century Plastics, 205 So. Puente St., Box 2393, Brea, CA 92622-2393, Tel: 1-800-767-0777; Fax: 1-800-786-7939, sell a wide variety of plastic sheets for jholding a wide variety of different size items. You might check with them to see if they have something to meet your needs. (They list stock No. V45000A-00 as having 4.5x5.25" pockets suitable for holding Polaroid prints.) Good luck,
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
Its interesting that you mention this. I never look forward to having to clean the tape off of my aluminum stages. The Acetone is labor intensive and time consuming. Just this morning I put a few mounts that had several layers of carbon tape on them and placed them in a platinum dish and set them in a muffle furnace -at- 300 C. After about an hour the tape had charred, but looked as though it was still going to take some scraping to get off. I increased the temperature to 400 C and left it for another hour. The tape had powdered and fallen off of the stages. I rinsed them in DI water and set them in an ultrasonic bath and they came clean.
Have you tried the commerial product "Goo-begon"? Its generally available in most N.American harware stores, painting stores, and grocery stores as a product to removing "Sticky residues left behind after removing tapes and labels".
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513-529-5712 Fax: 513-529-4243 E-mail: edelmare-at-muohio.edu
"WE ARE MICROSOFT. RESISTANCE IS FUTILE. YOU WILL BE ASSIMILATED."
R. Breeze and M. Turk; 1984; Cellular structure, function and organization in the lower respiratory tract; Environmental Health Perspectives 44:3-24, 1984.
J. A. Nowell and W.S. Tyler; 1971; Scanning electron microscopy of the surface morphology of mammalian lungs; Amer. Review of Respiratory Disease 103:313-328, 1971.
Also, look for more recent papers by C. Plopper, J. Crapo, or K. Pinkerton.
Jane A. Fagerland, Ph.D. Dept. Microscopy and Microanalysis Abbott Laboratories Abbott Park IL 60064
} What are some effective solvents for removing adhesive tape, spots, etc. } from aluminum SEM mounts. Acetone is marginal as are other ketones. } I've tried several alcohols without success. Caustics are out because } they eat the aluminum. Pure and mixed acids don't work either. I've } even tried WD-40 and liquid dish soap. } } The ideal solvent would have low toxicity, low cost, etc... Water } solubility would be a bonus. I'd really like a chlorinated solvent } vapor degreaser! Fat chance. } } Any suggestions? Terpenes, perhaps? } Dear Harold, I'm surprized that acetone and ethanol did not remove adhesive. The adhesive from tape in our lab is readily removed by these. I'd try an aromatic, preferrably xylene or toluene--used in a hood, of course. Avoid benzene--it's much more toxic than the hydroxylated aromatics. Freon is another possibility. I'd also avoid chlorinated solvents if possible because of toxicity. As you said, stay away from bases, including strong detergents. Fine abrasives are another option; something like wehnelt-polishing compound can be used. Good luck. Yours, Bill Tivol
} Does anyone know the dangers associated with a technique called Confocal } Microscopy or something like that? In our Chemistry Dept. we have a } research group that works with a Class 4 LASER. To the best of my } understanding, the LASER beam is directed toward a sample that is sitting } under a microscope. Some of the beam is reflected and filtered so the } power or strength of the beam is reduced before it hits the sample } (a biological sample). The researcher then looks through the microscope } objective at the sample. } } Currently, we have a LASER safety program in place. This particular } procedure concerns me because this technique allows the researcher to } look directly at the LASER beam. This does not sound too good. Also, } the microscope objective may magnify the beam and direct the beam } toward the researchers eye. } } Currently, the LASER that is being used is an Class 4 ARGON Ion LASER } (Continuous Wave) with a wavelength of 0.488 um (ultraviolet range) and a } beam power of 1.5 W. } } Does anyone know where I can get more information on this subject. I am } particularly interested in the safety aspects of this technique. It } may be the case that the LASER beam hitting the sample has a low enough beam } power to classify it as a Class 1 LASER. } } Thanks in advance for your help. } } Laurie Princiotto } Laboratory Safety Specialist } Indiana University - Dept. of Environmental Health and Safety } Phone: (812)-855-6115 } Fax: (812)-855-7906 } E-mail: lprincio-at-indiana.edu
Laurie, This does not sound like a very safe activity to me. I would be very concerned until proven safe.
Regarding the lasers used in confocal microscopes...I've worked with the Bio Rad commercial system and I believe most use a class II Ar or Ar/Kr laser at ~1mW power (457-648nm). The laser image cannot be directly observed but is displayed in a computer screen.
You're best info sources would be the manufacturers of confocal microscopes.
Edward J. Basgall, PhD The Pennsylvania State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618
My guess is that the citrus-derived "limonene" product used as a substitute for xylene in histology should work fine. It solubilizes paraffin, oils, etc. and is very safe. Most histo labs use this solvent these days, so drop by you local histo lab and give it a try.
} What are some effective solvents for removing adhesive tape, spots, etc. } from aluminum SEM mounts. Acetone is marginal as are other ketones. } I've tried several alcohols without success. Caustics are out because } they eat the aluminum. Pure and mixed acids don't work either. I've } even tried WD-40 and liquid dish soap. } } The ideal solvent would have low toxicity, low cost, etc... Water } solubility would be a bonus. I'd really like a chlorinated solvent } vapor degreaser! Fat chance.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 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 ####################################################################
Thanks to all who responded to my inquiry of March 19 concerning embedding and sectioning, I have summarized the responses below.
On the use of silane adhesion promoters (in my case Dow Corning Z-6040) for embedding in epoxy: Phil Swab uses these promoters on materials specimens. He uses 1% Z-6040 in 1:1 methanol:water for an hour then dries the specimen. He then embeds in epoxy with a replacement of half the amine polymerization initiator ( eg. DMP-30) with Z-6040. Stacie Kirsch of EMS, has used Z-6040 with epoxies and found no difference between conventional embedding and use of Z-6040. I ran an experiment using some bird feather barbs using EM-BED 612 (EMS) with the following protocols: In cases 1 & 2 acetone was the transitional solvent, and specimens were left in 1:1 acetone:epoxy overnight and in other acetone epoxy steps for a minimum of 4 hrs, and 4 hrs in the epoxy alone before transferring to the final epoxy embedment. 1. No Z-6040. 2. 1% Z-6040 added to all EtOH steps up through 100% EtOH but not in acetone. 3 Phil Swab method see above. To summarize, there were no differences in appearance under the beam. All blocks cut easily and equally well, and all suffered from the same problem. The problem was that on pickup on uncoated grids the feather sections fell out of the block. Mounting on coated grids was mandatory. If you use Z-6040, LRWhite appears to be the plastic of choice. If anyone has an experimental bent, trying some of the other adhesion promoting silanes may work. A long preembedding period in solvent+epoxy and in the pure epoxy certainly seems to help regardless of the other embedding details. Thanks to Caroline Schooley for suggesting Phil Swab.
Concerning cells on membranes: Neelima Shah suggested letting the membrane roll up (usually happens in higher alcohols or acetone in my experience) and vacuum infiltration. Wis Jablonski suggests using a hard Spurr mixture with long infiltration and the Spurr only step under mild vacuum. Karen Zaruba suggests the use of LRWhite and heat polymerization. Laura Patrone suggests cutting the block so that the cell layer is down (the first part of the specimen to be cut by the knife). Many of these correspondents gave suggestions for membrane inserts such as Millepore HA membranes, Millicel CM membranes. My problem was that I am restricted to the membranes being used by the investigators, and in the last batch I had, there was essentially no adhesion between the cell layer and the membrane, they split apart during the trimming (as soon as any shearing force was applied in the general vicinity). For the more tractable membranes the change in orientation of the block with respect to the knife seems a reasonable easy fix. Also longer infiltrations in plastic+ solvent and in plastic infiltration step work better. LRWhite is also worthy of consideration by the epoxyheads out there, such as myself.
I have no pecuniary connection to Dow Corning or EMS.
Bruce Cutler, Microscopy Laboratory, University of Kansas, Lawrence
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } What are some effective solvents for removing adhesive tape, spots, etc. } from aluminum SEM mounts. Acetone is marginal as are other ketones. } I've tried several alcohols without success. Caustics are out because } they eat the aluminum. Pure and mixed acids don't work either. I've } even tried WD-40 and liquid dish soap. } } The ideal solvent would have low toxicity, low cost, etc... Water } solubility would be a bonus. I'd really like a chlorinated solvent } vapor degreaser! Fat chance. } } Any suggestions? Terpenes, perhaps? } } } ------------------------------------------------ } Opinions or statements expressed herein, rational or otherwise, do not } necessarily reflect those of my employer. } } Harold J. Crossman } OSRAM SYLVANIA INC. } Lighting Research Center } 71 Cherry Hill Dr. } Beverly, MA 01915 } Phone: (508) 750-1717 } E-mail: crossman-at-osi.sylvania.com } } Our web sites: www.sylvania.com } www.siemens.com } -- } } "Crossman, Harold" {crossman-at-osi.SYLVANIA.com} } } "Crossman, Harold" {crossman-at-osi.SYLVANIA.com} } Harold, have you tried soaking them in a 10% solution of Microsolution? This has worked for me several times I've wanted to reuse stubs. Use hot water (the hotter the better) when starting then add the Microsolution. It works best with hot water but letting things soak in cold water overnight works O.K. too. Hope this helps.
Peace through Christ,
Phil Rutledge, Director e-mail: prutle1-at-gl.umbc.edu Center for Microscopy voice: (410) 455-3582 UMBC Dept. of Biology fax: (410) 455-3875 Catonsville, MD 21228 ///// / / / / /////// /////// / / /////// /////// / / / / / / / / / / / / /////
I may be dragging the thread away from contamination, but I thought that most electron microscope users avoided silicon based oils in diff pumps etc
because they are extremely difficult to remove from the interior of an electron microscope and any contamination would normally have electrical insulating properties (catastrophic in an e.m.). Perhaps I am wrong but I would welcome any comments. After all this is one of the reasons why Santovac oils and their relatives became so popular (despite their costs).
If I am labouring under a mis-apprehension then I apologise.
Malcolm Haswell University of Sunderland UK
All of the silicon based DP fluids I am aware of (which may not be all that are on the market) will break down under an electron beam and deposit a layer similar to glass on the nearest cool surface in the column. I don't know of any EM manufacturers that would recommend their use because they are almost impossible to remove if they do get in the column. We don't even use silicon based fluids in our vacuum evaporator.
Years ago I used to make carbonate-free NaOH by dilution of aliquots drawn from a saturated solution of NaOH pellets, the procedure came from the classic text on quantitative inorganic analysis by Vogel. The story is that Na2CO3 is more-or-less totally insoluble in saturated NaOH. I can't remember what concentration NaOH the saturated solution is, and, of course you have to dilute it with recently-boiled out water, but at least the problem of getting carbonate-free NaOH is thus eliminated.
cheers
Ritchie
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
Malcolm Haswell wrote: =============================================== .............. but I thought that most electron microscope users avoided silicon based oils in diff pumps etc because they are extremely difficult to remove from the interior of an electron microscope and any contamination would normally have electrical insulating properties (catastrophic in an e. m.). Perhaps I am wrong but I would welcome any comments. After all this is one of the reasons why Santovac oils and their relatives became so popular (despite their costs).
If I am labouring under a mis-apprehension then I apologise. ================================================= If you are laboring (sorry labouring) under a "mis-apprehension" then you are not alone!
The only time I have ever thought that the Dow Corning silicone based fluids would be acceptable for use in an EM lab was in the vacuum evaporator, unless of course you were interested in doing Si analyses by EDS and you did not want there to be even remotely the possibility of having Si contamination from the pump fluid. There are some who argue for other reasons even against the vacuum evaporator use.
But I have heard far too many horror stories over the years of columns being flooded with silicone DP fluid and then having the microscope undergo a month long or more cleaning job. Stick with a polyphenyl ether fluid, such as Santovac (R) or if you are really on a super tight budget, use dioctylsebacate (e.g. Octoil-S (R)) which are of course easy to clean up in the event of a column catastrophe.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
One of my colleagues is looking for the excitation and emission spectra for fluorogold. The supplier gives out the Ex & Em peaks but says they don't have a full spectrum. Has anyone out there generated their own? TIA.
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
We are preparing to purchase an ultramictrome for our centralized EM facility. This instrument will be used by a variety of people, including undergraduate students. If you have recently purchased such an instrument, we would appreciate your comments, good and bad. Pricing information would also be helpful.
Bill McManus Department of Biology Utah State University Logan UT 801 797 1920
Aloha! You can usually count on me to wade in with the High Humidity viewpoint...
We use Polaroid Type 55 positive/negative film, which requires the positives to be coated. In our usual high humidity environment the coating never really dries forever, and rehydrates easily. The coating on prints stored in various kinds of sleeves often ends up sticking to the material and, when the prints are peeled out, exhibits unsightly marks. I called Polaroid and got a list of acceptable materials that could come into contact with the coating and which included such things as ceramic and some plastics but not others. (Please call them rather than make me dredge the file cabinet for this list!).
I conducted a year-long experiment with prints stored in 3-ring storage sheets from various companies, made of various materials, left under stacks of heavy books. The sheets that work best under my conditions are Polyethylene Storage Sheets for 4X5 Negs and Prints, Order #4504, from
NegaFile P.O. Box 78 Furlong, PA 18925 215-348-2356
This is not to say that sheets from other sources won't be just fine for uncoated prints or coated prints in areas with lower humidity!
It's a beautiful, sunny day with a humidity of 70 percent IN THE LAB, and south shore surf is coming up.
Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
the response to my recent question about polaroid print storage was overwhelming. Thanks to those who responded I now have several lines of enquiry, even a couple in Australia. Cheers,
Mark Blackford TEM Group Materials Division, ANSTO PMB 1, Menai, N.S.W. Australia 2234 Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
the response to my recent question about polaroid print storage was overwhelming. Thanks to those who responded I now have several lines of enquiry, even a couple in Australia. Cheers,
Mark Blackford TEM Group Materials Division, ANSTO PMB 1, Menai, N.S.W. Australia 2234 Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
There is another option for scanning film negatives. A Leaf MicroLumina camera can be coupled with a high frequency light box and a copy stand to provide a simple and highly flexible solution to the problem of scanning film and transparencies.
The camera can be racked up and down on the stand to provide a field of view from 1" X 1.5" up to 9" X 12". The camera has a resolution of 2700 X 3380 pixels and can capture 12 bit grayscale and 36 bit color.
This configuration is currently offered by our company to the electron microscopy community under the name TEMSCAN.
If anyone is interested in this product, please contact us by phone: 516-773-4305 or e-mail: sales-at-electroimage.com.
Thanks to the several people who responded to my question.
The following was received directly from Bart Cannon and has some useful detail. I thought others might be interested.
} Arthur Wehnelt (1871-1944) was the German physicist who invented the } "Hot Cathode" electron tube which employed what was called a "grid" to } direct a stream of electrons. It corresponds fundamentally to the } electron gun used in oscilloscopes, TVs, and electron microscopes. Grid } and wehnelt have been interchangable terms to some degree.
-- Fred Schamber ....................... mailto:fhscham-at-SGI.NET
If you have "copies of copies" which work ok, and you have suitable blank media, there is no reason you cannot copy these again to generate archive disks. If you have a "verify" command/switch - use it. ...Or am I missing something??
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We have a NORAN (Tracor Northern) 5400 Series II EDS system (Model Number TN-5402/BBAA) acquired in 1987. It does not have a hard drive and operates off of two floppy drives. It requires 5.25" double sided, high density floppy disks, the originals of which are no longer readable. They have not been in use other than to be the source of copies made periodically but apparently they have simply deteriorated with time. We have copies of copies that are working pretty well but I'm becoming concerned that they may crash/die and we'll have no good source from which to make new copies. NORAN does not have replacement masters of this vintage so I'm hoping to find someone who has original master disks in good working order who would be willing to loan them to be copied.
We need two disks of the set from the November, 1987 Release 1C. What will work with our system is very specific and the original label on the master disks reads:
SERIES II SOFTWARE RELEASE 1C NOV., 1987, TRACOR NORTHERN, INC.
Of the disks in this release, we need:
SQ/SSQ/PRZ/ZAF Master NOT BOOTABLE
and
MSCAN2 Master
Thanks in advance for any help or suggestions you may have.
___________________________________________________________________________ Barbara Reine, Botany Dept. Box 351330 Univ. of Washington, Seattle, WA 98195-1330 e-mail: reine-at-u.washington.edu; ph: (206) 543-1955 ____________________________________________________________________________
While we're on the subject, and inspired by Chuck's comments about use of same in vacuum coaters, you may recall that about a year ago I asked about DC 704 in a coater used for carbon coating mineral samples for subsequent EDS analysis. I mounted up some graphite rod, analysed it
a uncoated b coated as usual ie with LN2 trap c as b, but left under diff pump vacuum for an extra hour d as c, but left under diff pump vacuum without LN2 in the trap for 2 hours(!). This was with a more-than-9-years-old charge of DC704 in the Edwards 306 coater.
Subsequent analysis showed no detectable Si in any of the samples.
So I cleaned out the pump, using Will Bigelow's favourite Tilex (thanks very much for the sample, Will) then Trichloroethylene, recharged it with DC704, and everything is hunky-dory.
In the diff pump of my JEOL JXA-5A, I use the JEOL-recommended Lion S oil, which I think is just a hydrocarbon oil, it's pretty cheap, and I get a vacuum better than 5 times 10 to the minus 6 torr. I change it every year.
cheers
Ritchie
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
Check with the Manufacturer of your SEM or the maker of your ODP for the proper oil. Hopefully it did not come from the factory with the silicone stuff. The type and quantity of oil should match the Pump. Typically Sanovac requires a hotter (higher Wattage) heater than Octoil. If you put Octoil in with a hotter heater you may have charcoal. And if you use a cooler heater with Sanovac you may never pump down.
It seems to me that if you have major back-streaming that you will see some oil elsewhere and be cleaning your column often to maintain decent astigmatism. I would recommend checking the detector window and eliminating the Silicone oil, before you have more problems.
I am not a chemist, but... I use Skelly B (i.e., hexanes?) in our lab to remove adhesive. I particularly use it to clean up after using pressure-sensistive adhesive polishing papers on our aluminum wheel. It is one of the few things that cut the adhesive at all.
At 11:42 AM 4/30/97 -0400, you wrote: } What are some effective solvents for removing adhesive tape, spots, etc. } from aluminum SEM mounts. Acetone is marginal as are other ketones. } I've tried several alcohols without success. Caustics are out because } they eat the aluminum. Pure and mixed acids don't work either. I've } even tried WD-40 and liquid dish soap. } } The ideal solvent would have low toxicity, low cost, etc... Water } solubility would be a bonus. I'd really like a chlorinated solvent } vapor degreaser! Fat chance. } } Any suggestions? Terpenes, perhaps? ---------------------------------------------------- Warren E. Straszheim 270 Metals Development, Ames Lab/ISU, Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-3091
Try an oily solvent such as Histolene. Its extracted from citrus and has a pleasant smell and is reputed to be of very low toxicity. Its sold as a substitute for xylene which is none of the above. OR eucalyptus oil. These are also good for getting bandaids off small children with a minimum of weeping.
I am seeking some input and suggestions on a problem that I am having with our Denton Desk II cold sputter coater.
About 2 weeks ago, a student came to inform me that the sputter button wasn't "popping" when depressed as usual - I believe the popping is the opening of the solenoid valve which admits Argon to the chamber. After confirming that the problem existed, I noted that the outlet valve from the regulator had been turned off (I'm not sure if this contributed to the situation or not). Upon opening the back of the unit, I found a blown 2 Amp slo-blow fuse.
After replacing the fuse, the sputter valve has been working just fine, however, after replacing it, a new problem arose - when the 45mA current was applied to the gold target (at 50 millitorr), the typical plasma glow was not evident even though the indicator read 45mA - I did notice some initial arcing, then nothing!
After consulting a very helpful sales/service representative (who suggested that I take the sputterhead apart and clean the teflon spacer/insulator) I was able to restore normal function - at least it seemed to be fixed for a few trial runs!
Now, no matter how many times I take the head apart and clean each piece, the coater fails after only a few runs or less - it arcs violently and the solenoid valve automatically shuts down (cycles to off position).
Has anyone had similar problems with the unit and/or any advise? This unit was purchased in 1992 and is not used excessively (1 to 2 SEM courses per year) - I recently replaced only my second gold target in that amount of time.
Thanks in advance for your kind assistance.
Stephen J. Beck Associate Professor Bio-Imaging Center/Electron Microscopy Department of Biology Nassau Community College Garden City, NY 11530 Voice Mail: (516) 572-7829 Email: {becks-at-sunynassau.edu} URL: {http://www.sunynassau.edu/webpages/biology/becks.htm}
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We have recommended Ross Ultra Super Glue to our customers as the best glue to use for Tripod Polishing applications. Recently, Ross has "improved" their formula and it is no longer satisfactory.
This note serves 2 purposes:
1) As a warning to Tripodders to beware of the "new and improved" Ross Ultra Super Glue (Blue label)
2) To ask if anyone out there has found another suitable glue to use. The main concern is for wedge polishing applications where the sample will be reduced to only a few microns in thickness.
There are super glues out there that work fine, but we are on a quest to find the best. Any help you can provide would be great!
David Henriks TEL: 800-728-2233 (toll-free in USA) South Bay Technology, Inc. 714-492-2600 1120 Via Callejon FAX: 714-492-1499 San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } What are some effective solvents for removing adhesive tape, spots, etc. } } from aluminum SEM mounts. Acetone is marginal as are other ketones. } } I've tried several alcohols without success. Caustics are out because } } they eat the aluminum. Pure and mixed acids don't work either. I've } } even tried WD-40 and liquid dish soap. } } } } The ideal solvent would have low toxicity, low cost, etc... Water } } solubility would be a bonus. I'd really like a chlorinated solvent } } vapor degreaser! Fat chance. } } } } Any suggestions? Terpenes, perhaps? } }
I have been using petroleum ether with considerable success. Used in a fumehood it is quite safe on aluminium, most plastics and other surfaces such as glass, wood and even paper.
-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.-.- Glynis de Silveira University of Cambridge Department of Materials Science and Metallurgy E-m:gds1002-at-cam.ac.uk Pembroke Street, UK Tel:+44(0)1223 334434 CB2 3QZ Fax:+44(0)1223 334567
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have bought a AGFA DUOSCAN scanner recently, it has a tray for positives and one for negatives up to A4 size (~8"X11")
Resolution is 2000x1000dpi optical (4Kx4K interpolated but who would use that!).
Works fine for me but I find it rather slow!
Usual disclaimer apply, I have no connection with AGFA I am only a customer...
Jean-Marc Boichat email: jean-marc.boechat-at-chma.mhs.ciba.com EM LABS FO 5.1 phone:+4126 435 6979 fax: +4126 435 6907 Ciba research Center P.O. Box 64 CH- 1723 Marly 1 When things go wrong, don't go with them! Switzerland
Disclaimer: "nobody in this company ever cared for what I said, why would they start now".
X400-Received: by host241.abbott.com (Internal Mail Agent-1); Thu, 1 May 1997 08:27:29 -0500 Alternate-Recipient: prohibited
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
On Wed, 30 Apr 1997, Crossman, Harold wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } What are some effective solvents for removing adhesive tape, spots, etc. } from aluminum SEM mounts. Acetone is marginal as are other ketones. } I've tried several alcohols without success. Caustics are out because } they eat the aluminum. Pure and mixed acids don't work either. I've } even tried WD-40 and liquid dish soap. } } The ideal solvent would have low toxicity, low cost, etc... Water } solubility would be a bonus. I'd really like a chlorinated solvent } vapor degreaser! Fat chance. } } Any suggestions? Terpenes, perhaps? } } } ------------------------------------------------ } Opinions or statements expressed herein, rational or otherwise, do not } necessarily reflect those of my employer. } } Harold J. Crossman } OSRAM SYLVANIA INC. } Lighting Research Center } 71 Cherry Hill Dr. } Beverly, MA 01915 } Phone: (508) 750-1717 } E-mail: crossman-at-osi.sylvania.com } } Our web sites: www.sylvania.com } www.siemens.com } -- } } "Crossman, Harold" {crossman-at-osi.SYLVANIA.com} } } "Crossman, Harold" {crossman-at-osi.SYLVANIA.com} } Harold, have you tried soaking them in a 10% solution of Microsolution? This has worked for me several times I've wanted to reuse stubs. Use hot water (the hotter the better) when starting then add the Microsolution. It works best with hot water but letting things soak in cold water overnight works O.K. too. Hope this helps.
Peace through Christ,
Phil Rutledge, Director e-mail: prutle1-at-gl.umbc.edu Center for Microscopy voice: (410) 455-3582 UMBC Dept. of Biology fax: (410) 455-3875 Catonsville, MD 21228 ///// / / / / /////// /////// / / /////// /////// / / / / / / / / / / / / /////
If you are not concerned with a smooth surface Al stub, I just grind off the samples on a piece of fine sandpaper, dump these into acetone and let it sit for several hours. Pour off dirty acetone, add clean acetone and then pull out stubs and let dry on paper towels. I normally process about 30 or so stubs at a time. Most of my users take the stubs with them, so I don't have to do this often. Bruce Cutler, Microscopy Lab, Univ. Kansas, Lawrence
} Harold, } have you tried soaking them in a 10% solution of Microsolution? } This has worked for me several times I've wanted to reuse stubs. Use hot } water (the hotter the better) when starting then add the Microsolution. } It works best with hot water but letting things soak in cold water } overnight works O.K. too. Hope this helps. }
I have used microsolution to clean vacuum parts, mainly stainless steel. I have found that it severly attacks aluminum parts, therefore, I no longer use it for aluminum.
Michael T. Marshall Research Engineer, Electron Microscopy University of Illinois at Urbana-Champaign Frederick Seitz Materials Research Laboratory 104 South Goodwin avenue Urbana, IL 61801-2985 (217) 244-8193 fax: (217) 244-2278
Harold, The adhesives we usually encounter in microscopy are probably at least 60% cross-linked so there is no magic, modest hazard, solvent that will fully dissolve them. - Heptane will swell some adhesives and then a heptane wetted paper towel will wipe the adhesive off the mount. - THF will dissolve the non cross-linked component of some adhesives and that could make the adhesive easier to remove. - We use acetone, ethanol and muscle.
As always, read the MSDS for the solvent and obey the numerous, frequently changing, local, state and federal regulations for transportation and disposal.
...the usual disclaimers...At 11:42 AM 4/30/97 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Yes, one more request for info on the best camera to use with our microscope-computer set up. I'm sorry for not keeping all the info that has been out on this subject a few months back, but I thought there was no way the money would show up for one any time soon. Wrong. We have an Olympus BH microscope and a Power Mac computer. We are interested in using the NIH image software with this set up, but we need a CCD camera with at least good resolution and any software/hardware to link the camera and computer. Notes on personal experience with these systems would be appreciated. I will need to know how much the camera and interface will cost too. Once again, sorry for not paying closer attention the first time.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
In response to Mr. Henriks 'quest' for better Super-Glues for Wedge Polishing of TEM samples:
Through a bit of research on the subject, it has become apparent that the cyanoacrylate adhesives used for Wedge Polishing in general are not capable of forming a strong chemical bond to the surface of glass. This is due mainly to their affinity for materials to which water will adsorb. (Thus the problem with 'Super-Gluing' one's fingers together.) Since the majority of vendors selling 'Tripod'-like polishing fixtures for wedge polishing also supply only glass or PYREX stubs for mounting the sample, it is no surprise that the samples are often lost after hours of effort has been expended on thinning the sample.
For years, wedge polishers have been switching from one cyanoacrylate product to the next in order to find one that will adhere better to glass. Two years ago, after looking into the problem, it became clear to me that using a transparent polymer instead of glass for the mounting stub would solve the problem. The trick was to find a polymer that could withstand the attack of cyanoacrylate solvents such as acetone and nitroethane. The solution made the choice of cyanoacrylate adhesive much less important.
As this note is meant to be informative as opposed to an advertisement for product, I would be happy to discuss our solution to the 'Super-Glue' problem in more detail off-line.
Regards, Scott D. Holt BUEHLER LTD. 41 Waukegan Rd. Lake Bluff, IL 60044 USA (847)295-4546 Fax (847)295-7942 http://www.buehlerltd.com
We are looking for SEM slow scan image digitizer and TEM image digitizer - for really old microscopes: Philips SEM 503 and TEM 300. We would appreciate any comments, including pricing information.
Vladimir Dusevich Temple University dusevich-at-astro.ocis.temple.edu
The laser is exciting fluorescence in the sample. This is what is being viewed just as we view blue light when exciting with UV using fluorescence stains such as DAPI. We just went through establishing a laser saftey program on our campus. The saftey dept. determined that if all equipment using lasers for visual inspection of samples had saftey features built in such that the user could not be exposed to the laser. The notable exception was during service.
Tommy Sewall
} } } "Frieda Christie" {f.christie-at-rbge.org.uk} 04/30/97 04:10pm } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Forwarded message:
I use ethanol and sonicate for awhile. Sometimes the stubs need to be rubbed on a paper towel even after this treatment, but the adhesive is fairly easily removed in this way. But I agree, it would be nice to have something that would miraculously make the adhesive instantly dissolve! Regards, Melanie Behrens
If I am off-base with this response, I apologize. I have used nothing but carbon tape ever since it became readily available through regular EM suppliers but I always dreaded having to clean it off my sample holders because it was so sticky and tore so easily. Being a simple minded person, I just took a 3/8" wooden dowel, sharpened one end(like a chisel) and used it to roll the tape up into a wad that I could remove with my fingers. All the adhesive seems to comes off with the tape, but if any should be left behind it should easily come off with a mild solvent. (I always polish my holders, so I'm not sure about residual adhesive.) I also cut a piece of latex tubing to fit on the end of the rod to cushion my hand. This works for the carbon tape, I'm not sure about the other types of adhesive tapes. For what it's worth.
Bill -- ============================================================= Bill Chissoe III Electron Microscopist,University of Oklahoma E-mail: wchiss-at-ou.edu Ph. (405)325-4391 =============================================================
There are are few 3rd party organization that service Tracor/Noran Equipment.
We use Evex Analytical to service our equipment. They are affordable and provide prompt service. The Evex Headquarters is in Princeton, NJ and have field engineer located throught the country. They can be reached at 609-252-9192.
With reference to Holt's response to Henriks' post: We prep over 1500 specimens per year and have used Ross superglue, a cyanoacrylate, for years with no problem until they recently changed their formulation. The old Ross bonded very well with glass. We have been evaluating replacements and have no recommendation at this time (Read: everything we tried so far s%#*s!).
Scott: If you have a replacement that works, share it on line. I'm sure you can tell us something and not make it sound commercial!
Please contact Scion Corporation or go through their web site. http://www.scioncorp.com/
On Thu, 1 May 1997 kna101-at-utdallas.edu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hi all, } } Yes, one more request for info on the best camera to use with our } microscope-computer set up. I'm sorry for not keeping all the info that } has been out on this subject a few months back, but I thought there was } no way the money would show up for one any time soon. Wrong. } We have an Olympus BH microscope and a Power Mac computer. We } are interested in using the NIH image software with this set up, but we } need a CCD camera with at least good resolution and any software/hardware } to link the camera and computer. Notes on personal experience with these } systems would be appreciated. I will need to know how much the camera and } interface will cost too. } Once again, sorry for not paying closer attention the first time. } } Karen }
Tseng-Ming Chou (Alex) Dept. of Materials Science and Engineering Stevens Institute of Technology Castle Point on Hudson, Hoboken, NJ 07030 e-mail: tchou-at-attila.stevens-tech.edu tchou-at-menger.eecs.stevens-tech.edu The Microstructure Group of Stevens
Hello, I need your assistance in acquiring several cell cross sections prepared by ultra-microtomy. It is my hope that some of our BIO-TEM friends have specimens on grid that are no longer of interest. We are not particular as to the type of cell but for reference would like to know what it is & the basic prep. technique. Anything taht has been=20 viewed in a TEM would be fine. Contrast enhancement technique are not required. Carbonless grids might be preferable. Our EM group consist of Material Scientist, Nano Particle Chemist & Physical Electronics Scientist, thus we lack the equipment & skills to do this our selves so we=92re bumming.=20 The application is 109nm (VUV) holography. We will gladly take care of expenses incurred on our behalf.
Thank you in advance for any anecdotal experiences with the various visible and fluorescent image capture and archiving systems available. I am looking for a combination of camera/video board/storage media/software for organizing images that will allow long term indexable storage of high resolution images. I have tried to look through the archives of the listserver for the past two months, but didn't find anything about this (but did find out a lot about Bremstrahlung).
Sincerely,
G. Steven Bova Departments of Urology and Pathology Johns Hopkins University
I need information about the basic principles and applications of the different microscopy techniques ( LM, SEM and TEM ) applied to the characterization of pure metals (like copper).
I need answers to simple questions like: What kind of information can I get with each technique? What makes one better than another and for what purposes? How to decide which one to choose for characterizing a pure metal like copper?
I would appreciate very much any help or reference that you could give me. Thanks.
Addendum to this message sent a few hours ago: I need to use a PC platform- Thanks to Michael Shaffer for pointing out this omission.-GSB
Dear All,
Thank you in advance for any anecdotal experiences with the various visible and fluorescent image capture and archiving systems available. I am looking for a combination of camera/video board/storage media/software for organizing images that will allow long term indexable storage of high resolution images. I have tried to look through the archives of the listserver for the past two months, but didn't find anything about this (but did find out a lot about Bremstrahlung).
Sincerely,
G. Steven Bova Departments of Urology and Pathology Johns Hopkins University
I've always just scraped off the old specimen and sonicated the stubs in acetone or ethanol. If all of the crud isn't removed, it also doesn't usually matter, since it is covered up by any new tape/silver or carbon paint and the specimen.
For analytical WDX or EDX, then, yes, removing the tape, adhesive, and a few layers of aluminum is useful.
Phil
&&& Illigitimi non carborundum &&&&&&&& Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217) 355-1143 oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
I am sorry, but the stock number I gave in my previous comments about plastic pages sold by 20th Century Plastics Co to hold Polaroid prints is for those made of vinyl, which are not recommended for archival storage. The corresponding pages made of polypropylene, which are recommended for archival storage, have stock number EZV450A-00. Each page has four pockets suitable for holding 4.25" x 5.25" prints, giving a capacity of 8 prints (back to back) per page. The holes for the binder rings are in a tab that runs along the side of the page, completely outside the pockets, thereby eliminating any need for having to punch holes in the prints. I have no commercial interest, just trying to set the record straight.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
Advanced Imaging Concepts, Inc. provides just such systems, we produce the databasing & archiving software "Image Central" and supply that as just software, sell it with a frame grabber board or supply complete turnkey systems for archiving, or full blown image networks, or enable existing networks. We handle a large array of cameras, printers, grabbers, and image analsysis software to complete your digital workstation please contact me at AIC I will be glad to discuss your requirements with you and set up a demonstration if desired.
Scott E. Berman Sales Manager Advanced Imaging Concepts, Inc. Princeton, NJ phone(609) 921-3629 x26 fax(609) 924-3010 email Scott E57-at-aol.com
We use Rubber Cement Thinner to remove adhesive. I don't know what's in it but it is readily available and more effective than single solvents.
Jacob Bastacky, M.D. Room 116 Donner Laboratory Lawrence Berkeley Laboratory EMail: sjbastacky-at-lbl.gov University of California Phone: (510) 486-4606 Berkeley, California 94720 Fax: (510) 486-4750
Negafile has polaroid 4x5 pages 4 per sheet. I'm not sure if they have gone to one size now or if they still have the two 4x5 sizes. You used to have to specify an "L" in the part number when ordering, but the number on the sheet itself, 4504, was the same for both. Check with them. I do not have their phone number, but their address is P.O. Box 78, Furlong, PA 18925, USA
- -Scott Walck
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
POST-DOCTORAL RESEARCH FELLOW Princeton University
A post doctoral research fellow position is available in the Princeton Materials Institute and the Princeton Center for Complex Materials at Princeton University. The appointment is for one year initially with the possibility of renewal. It involves the use of a Philips CM-200 FEG transmission electron microscope equipped with a Gatan Image Filtering System in studying carbon nanotubes, and other nanostructured materials of interest. Candidates should have a Ph.D. in the physical sciences, and with strong EM background (HREM, Electron diffraction, and EELS). Experience in image analysis and UNIX workstations, and a strong math background are highly desirable.
Applicants should send a detailed curriculum vitae, copies of 2 selected publications, and names and addresses of three referees by June 15, 1997 to:
Nan Yao Princeton Materials Institute Princeton University Bowen Hall, 70 Prospect Avenue Princeton, NJ 08540
Princeton University is an equal opportunity employer.
=============================================================== Nan Yao Princeton Materials Institute Princeton University Bowen Hall, 70 Prospect Ave. Princeton, New Jersey 08540-5211
Hello, I need your assistance in acquiring several cell cross sections prepared by ultra-microtomy. It is my hope that some of our BIO-TEM friends have good cell sections on grid that are no longer of interest. We are not particular as to the type of cell but for reference would like to know what it is & the basic prep. technique.=20 Anything that has been viewed in a TEM would be fine. Contrast enhancement technique are not required. Carbonless grids might be preferable. Our EM group consist of Material Scientist, Nano Particle Chemist & Physical Electronics Scientist, thus we lack the equipment & skills to do this our selves so we=92re bumming.=20 The application is 109nm (VUV) holography. We will gladly take care of expenses incurred on our behalf.
Just a few more comments in an attempt to answer questions that have been raised about the characteristics of different diffusion pump fluids and their use in electron beam systems (matters which are discussed in some detail in Sect. 5.4 of my book, "Vacuum Methods in Electron Microscopy").
We ran into the problem of identifying Lion-S oil a number of years ago, and finally decided that it is most probably the synthetic fluid di(2-ethyl-hexyl) sebacate, and not a hydrocarbon oil. This same compound is sold as a pump fluid by several other companies under various trade names, most of which contain the letter S, such as Octoil-S (CVC) Diffoil-S (Lesker), etc. It has a vapour pressure at 20=B0C of about 3 x 10-6 Pa, and a boiling point at the normal boiler pressure of diffusion pumps (about 100 Pa) of about 205=B0C. It has better resistance to thermal and oxidative degradation than hydrocarbon oils, but will still break down if exposed to air at pressures much above 100 Pa while at operating temperature. We had trouble obtaining Lion-S oil when we serviced the DP on one of our SEMs and so used Octoil-S in its place, and had no problems as a result.
DC-704 is a synthetic silicone compound (tetraphenyl-tetramethyl-trisiloxane) that has a vapour pressure at 20=B0C o= f about 3 x 10-6 Pa and a boiling temperature at 100 Pa of about 220=B0C. Thu= s the properties of DC-704 are so nearly the same as those of di(2-ethyl-hexyl) sebacate that a pump designed for one fluid will work quite well with the other, without modification of heater wattage. The silicone fluids are very resistant to thermal, oxidative, and radiative degradation, hence their great advantage in a number of industrial processes. As pointed out by others, however, under electron bombardment they break down to form non-conducting siliceous deposits which are very insoluble, and so are not normally used in electron microscopes these days. I remember when RCA used silicone oils in some of their early microscopes. These instruments had platinum apertures, and in order to clean the aertures we had to flame them first (removing the organic component of the contamination) and them soak them in hydrofluoric acid to get rid of the siliceous residue.
The level of oil contamination produced by an diffusion pump depends not only on the type of pump fluid used, but also critically on the design of the system, and of the pump itself (see. Vac Meth p. 190-198). Several manufacturers of diffusion pumps devoted a tremendous amount of effort to developing pump designs that minimize backstreaming, and so systems equipped with their pumps will give minimal trouble in this regard. However, early models of diffusion pumps did not incorporate these critical design improvements, and so if you happen to have a system equipped with one of these pumps you can expect lots of oil in your system. Also, it seems that in the 'early days' some manufacturers of electron microscopes made their own diffusion pumps, and , of course, these often produced high levels of contamination, because they did not incorporate the design features needed to reduce backstreaming. The use of cold caps, water-cooled baffles and liquid nitrogen traps above the diffusion pumps will also reduce contamination rates (Vac Meth p. 190-198). The problem is, the design of the pump and its associated accessories is pretty much set by the manufacturer, and so it is rather difficult for the user of an instrument to do much about its basic characteristics. However, there are several mishaps that can occur during the operation of a diffusion pump (i.e. loss of cooling water, failure of the backing pump, exposure to air at high pressure, loss of electric power) which can lead to severe degradation of the pump oil (e.g. see Fig. 5.15 Vac Meth, p 218) and produce horrendous levels of backstreaming (Vac Meth , p. 214-223), and users should be extremely carful to guard against the occurrence of any such events.
=46inally, as I mentioned before, experience shows that in many cases the major source of oil contamination in systems with oil diffusion pumps is the breakdown of the hydrocarbon oil in the rotary vane backing pump (Vac Meth p. 144)
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
} Date: Thu, 1 May 1997 10:45:13 -0400 (EDT) } From: Vladimir Dusevich {dusevich-at-astro.ocis.temple.edu} } To: microscopy-at-Sparc5.Microscopy.Com } Subject: Image digitizers
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear Colleagues } } We are looking for SEM slow scan image digitizer and TEM image digitizer - } for really old microscopes: Philips SEM 503 and TEM 300. We would } appreciate any comments, including pricing information. } } Vladimir Dusevich } Temple University } dusevich-at-astro.ocis.temple.edu
A possibility for the SEM is a device called ImageSlave sold in Australia by OED (Fax +61 (0)2 9482 1196) I understand this has been fitted to a number of older SEM's around the world
Ian
(I have no financial interesed in OED or ImageSlave)
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-849 3660 EMail ihallett-at-hort.cri.nz
With the recent traffic concerning uranyl acetate and lead citrate, it seems an appropriate time to ask a question that has puzzled me for some time. It concerns using Uranyl acetate as an en bloc stain. Protocol for en bloc staining vary. Some people use 2% UA in water after osmium postfixation. Others do the UA block stain during the dehydration (ie; alcoholic UA) while yet another group go to the trouble of making up a Maleate buffered UA block stain. Maleate buffered UA block stain seems to be the most commonly recommended one, yet I have not been able to find out why it is preferred to the others. Can anyone suggest why Maleate buffered UA is preferred over both aqueous and alcoholic UA when it is used as an enbloc stain?
Thanks in advance,
Allan Mitchell, Technical Manager South Campus EM Unit Otago School of Medical Sciences Dunedin New Zealand
---------- } From: Steve Beck {becks-at-sunynassau.edu} } To: Microscopy-at-sparc5.microscopy.com } Subject: Denton Desk II Sputter Coater } Date: Thursday, May 01, 1997 12:09 AM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear Colleagues, } } I am seeking some input and suggestions on a problem that I am having with } our Denton Desk II cold sputter coater. } } About 2 weeks ago, a student came to inform me that the sputter button } wasn't "popping" when depressed as usual - I believe the popping is the } opening of the solenoid valve which admits Argon to the chamber. After } confirming that the problem existed, I noted that the outlet valve from the } regulator had been turned off (I'm not sure if this contributed to the } situation or not). Upon opening the back of the unit, I found a blown 2 Amp } slo-blow fuse. } } After replacing the fuse, the sputter valve has been working just fine, } however, after replacing it, a new problem arose - when the 45mA current } was applied to the gold target (at 50 millitorr), the typical plasma glow } was not evident even though the indicator read 45mA - I did notice some } initial arcing, then nothing! } } After consulting a very helpful sales/service representative (who suggested } that I take the sputterhead apart and clean the teflon spacer/insulator) I } was able to restore normal function - at least it seemed to be fixed for a } few trial runs! } } Now, no matter how many times I take the head apart and clean each piece, } the coater fails after only a few runs or less - it arcs violently and the } solenoid valve automatically shuts down (cycles to off position). } } Has anyone had similar problems with the unit and/or any advise? This unit } was purchased in 1992 and is not used excessively (1 to 2 SEM courses per } year) - I recently replaced only my second gold target in that amount of } time. } } Thanks in advance for your kind assistance. } Dear Steve,
We had the same problem with our Denton Desk II, to the extent that the gold foil target was torn by the arcing. We "solved" our problem by increasing the argon pressure to 70 millitorr during sputtering. This does not appear to have had any adverse effects on the quality of the coating and has eliminated the arcing problem completely.
At HRL we are urgently seeking a circuit board for the Motorised Stage Drive (ISM-MSD40-2) for our Jeol 840 SEM. The required circuit board is the computer interface (RS232) for external control, ie a EDXA control of the SEM stage. Should anyone know the whereabouts of a spare board or can get the schematic of this board we will be glad to here from you.
Warren Straszheim wrote: ================================================ What are some effective solvents for removing adhesive tape, spots, etc. from aluminum SEM mounts. Acetone is marginal as are other ketones. I've tried several alcohols without success. Caustics are out because they eat the aluminum. Pure and mixed acids don't work either. I've even tried WD- 40 and liquid dish soap. ================================================ Even though we have manufactured SEM mounts for others for more than 25 years, we ourselves have always had a program of cleaning and recycling our own mounts used internally. What you end up using depends a lot on just how much elbow grease you are willing to expend at the beginning in terms of removing the material physically. And Andy Blackwood in our lab just told me, some times they "give the mounts a 'lick' on 600 grit on the metallographic wheel, thus removing the whole surface."
But what ever solvents are eventually used, the real question is, just how many "rinsings" are enough in order to reduce the level of organics on the surface to an "acceptable" level. And I have not ever figured out how to make that determination.
Therefore, for critical samples, requiring the highest performance, samples that might not be receiving any further gold or carbon coatings, for example , we expose the mounts for a few minutes to an oxygen plasma in our SPI Plasma Prep II plasma etcher. Call it "etching" or call it "plasma cleaning", but in order to be absolutely sure there are no remaining organic residues, this is the only method literally guaranteed to remove the last remains of any organic residues.
PS: Even brand new SEM mounts, while they look and feel brand new, for high performance work, not other wise using applied conductive coatings, should be given some kind of a cleaning, the best one being this kind of a plasma cleaning treatment.
Information about plasma etchers for this kind of cleaning can be found on our website given below. We think that recycling mounts in this kind of situation makes a lot of good sense, provided the time, labor, and other wise unnecessary exposure to solvents exceed the benefits from the recycling .
Disclaimer: SPI Supplies manufactures the SPI Plasma Prep II plasma etcher and has a vested interest in seeing more uses for this system!
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I have had a query from a collegue who wonders how much success people have had running MAC versions of NIH Image with the PC MAC emulator Executor. I would appreaciate any comments and/or contact details of anyone with experience of this combination.
Thanks in Advance
Ian
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-849 3660 EMail ihallett-at-hort.cri.nz
Dear Harold and all, I have found that the best way to remove the carbon adhesive discs from Al stubs is to soak the stubs in acetone for at least 1/2 hour, then sonicate. When you rub the wet stub on a paper towel, the disc will slip off. Methanol is my solvent of choice for tape residues, but often works poorly if the residue is old. Most of the rubber cement-type glues respond to chloroform. My $0.02CAN ($0.014US) worth. Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
Dear All, I find oil on the EDX detectors in both my SEM's, but if you think of the conditions inside the SEM, it is almost inevitable. The majority of the molecules remaining in the vacuum of an average SEM at 10 -5 torr consists of diffusion pump oil. This oil will gradually condense on all surfaces in the SEM, but on the coolest surface first. In most SEM's this is the EDX snout. One EDX manufacturer has solved the problem by warming up the snout with a small heater. The oil just goes somewhere else, but if it forms a thin film over all surfaces it is not so visible or worrying. The cleaning method recommended by the manufacturer of my EDX's is to gently run clean-grade iso-propanol over the snout, then allow to air-dry. They used to recommend Freon, but that is no longer permitted or available. Hope this helps, Mary
Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear colegues, I found organelles that look like clathrin coated vesicles in the location where for some reasons coated vesicles should not be. Do somebody know about other organelles that have similar structure? I'll appreciate any information, Thank you in advance, Elia
Elia Beniash, PhD Student, Dept of Stuctural Biology, Weizmann Institute of Science, 76100, Rehovot, Israel
Hi, We are considering the purchase of an processor for TEM samples. We currently process about 10 to 15 samples a week. Would individuals please recommend or discuss with us their experiance using these instruments. Thanks, Robert Cox Shriners Burn Institute Galveston Tx, rcox-at-sbi.utmb.edu or 409-770-6655
I am interested in buying a color camera for phase, brightfield, and fluorescence imaging, which will be our only camera. At this point I don't absolutely need to do any ratioing or math involving intensities. I am interested in low noise but probably don't need exposures longer than a couple seconds (in fact motion of my wet-mounted samples may limit exposure time). I want to focus and make exposure adjustments in near-real time from the computer or video screen, not through a viewfinder. Expense is a consideration, too. With our currrent cheap monochrome video camera (Sony XC-75, TCX frame grabber) we have plenty of illumination but still lots of noise.
Are there any good systems (camera+grabber) that give greater than 8 bits per color after digitization?
The Diagnostic Instruments Spot camera is a 24 bit cooled ccd camera using a single Kodak KAF1400 chip. It uses some type of switchable liquid crystal filters to take 3 sequential images, one each RGB. Has anyone had experience with it, and compared it to the alternatives? I expect that due to the filters it is significantly less sensitive than others, is that true? How does noise compare with images digitized from color video cameras, cooled or not? Are there any other considerations?
For video cameras, what frame grabbers/drivers give histograms of light intensity to aid in optimizing exposure settings?
Thanks for the prompt delivery of the last order. I need to make another now. I need 2 #PA0050 It is a tonor cart. for a Lexmark Optra-R printer( a hateful machine). The price at the last order was $165 but if that has changed it is OK just e-mail me back the price. I need this billed out as soon as you can even if the items are not available. The blanket PO's need to be closed out for the fiscal year here soon. I can wait on delivery. Thanks mucho!!
PO# L06681 cust. # 6646947
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 352-392-1295 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
} } } } } ------- Forwarded Message Follows ------- } } } } } Does anyone know the dangers associated with a technique called Confocal } } } Microscopy or something like that? In our Chemistry Dept. we have a } } } research group that works with a Class 4 LASER. To the best of my } } } understanding, the LASER beam is directed toward a sample that is sitting } } } under a microscope. Some of the beam is reflected and filtered so the } } } power or strength of the beam is reduced before it hits the sample } } } (a biological sample). The researcher then looks through the microscope } } } objective at the sample. } } } } } } Currently, we have a LASER safety program in place. This particular } } } procedure concerns me because this technique allows the researcher to } } } look directly at the LASER beam. This does not sound too good. Also, } } } the microscope objective may magnify the beam and direct the beam } } } toward the researchers eye. } } } } } } Currently, the LASER that is being used is an Class 4 ARGON Ion LASER } } } (Continuous Wave) with a wavelength of 0.488 um (ultraviolet range) and a } } } beam power of 1.5 W. } } } } } } Does anyone know where I can get more information on this subject. I am } } } particularly interested in the safety aspects of this technique. It } } } may be the case that the LASER beam hitting the sample has a low enough beam } } } power to classify it as a Class 1 LASER. } } } } } } Thanks in advance for your help. } } } } } } Laurie Princiotto } } } Laboratory Safety Specialist } } } Indiana University - Dept. of Environmental Health and Safety } } } Phone: (812)-855-6115 } } } Fax: (812)-855-7906 } } } E-mail: lprincio-at-indiana.edu } } } } Laurie, } } This does not sound like a very safe activity to me. I would be very } } concerned until proven safe. } } } } Regarding the lasers used in confocal microscopes...I've worked with the } } Bio Rad commercial system and I believe most use a class II Ar or Ar/Kr } } laser at ~1mW power (457-648nm). The laser image cannot be directly } } observed but is displayed in a computer screen. } } } } Your best info sources would be the manufacturers of confocal microscopes. } } } } Leica - Ph: 415-348-2233 } } Bio-Rad - Ph: 800-4BIORAD } } Molecular Dynamics - http://www.mdyn.com ph:800-333-5703 or 408-773-1222 } } Zeiss 800-233-2343 } } } } good luck } } } } Edward J. Basgall, PhD } } The Pennsylvania State University } } Surface Chemistry Group ejb11-at-psu.edu } } Materials Research Institute Building Ph: 814-865-0493 } } University Park, PA 16802-7003 FAX: 814-863-0618
Laurie,
I have never heard of a laser confocal microscope that involves direct observation. To my knowledge, all of the laser confocal microscopes are scanning devices and the scan-derived image is displayed on a monitor -- the user never looks directly into the microscope.
Since you did not identify the make or model of the confocal scope, it is not clear to me that you have even spoken to the users about your safety concerns. That is generally a good place to start. They may know more about laser safety than you give them credit for. After that, talk to the company that makes the system. After that, send up a posting to the list with enough relevant background information, and I am sure you will receive informative responses.
As far as whether the laser is Class 4 or Class 1, it is not a good idea to look directly at the beam of any laser. Just in case anyone asks you that, in your capacity as a safety officer.
Duane McPherson Dept. of Biology SUNY at Geneseo Geneseo, NY
Advanced International Immunofluorescence Course Gargnano '97 (Italy)
The Advanced International Immunofluorescence Course is a post-doctorate theoretical/practical course, with propedeutical lectures and practical stages on traditional and confocal immunofluorescence microscopy and image and ion analysis. The course will take place in Gargnano (Lake of Garda) from 7 to 10 October 1997. Further information and registration details will be found at the following Web address
http://imiucca.csi.unimi.it/endomi/ACIF.html
Program:
Morning Tuesday, October 7, 1997
9.00-10.00 Registration of partecipants
10.00-10.45 AN INTRODUCTION TO IMMUNOCYTOCHEMISTRY AND IMMUNOFLUORESCENCE (I.Barajon)
10.45-11.15 Coffee Break
11.15-12.00 LIGHT SOURCES AND FLUOROCHROMES (G. Bottiroli)
12.00-12.45 THE USE OF FLUORESCENCE MICROSCOPE (C.Rumio)
Advanced International Immunofluorescence Course Gargnano '97 (Italy)
The Advanced International Immunofluorescence Course is a post-doctorate theoretical/practical course, with propedeutical lectures and practical stages on traditional and confocal immunofluorescence microscopy and image and ion analysis. The course will take place in Gargnano (Lake of Garda) from 7 to 10 October 1997. Further information and registration details will be found at the following Web address
http://imiucca.csi.unimi.it/endomi/ACIF.html
Program:
Morning Tuesday, October 7, 1997
9.00-10.00 Registration of partecipants
10.00-10.45 AN INTRODUCTION TO IMMUNOCYTOCHEMISTRY AND IMMUNOFLUORESCENCE (I.Barajon)
10.45-11.15 Coffee Break
11.15-12.00 LIGHT SOURCES AND FLUOROCHROMES (G. Bottiroli)
12.00-12.45 THE USE OF FLUORESCENCE MICROSCOPE (C.Rumio)
There are many issues here when making a choice on a CCD camera, among which are: 1) color vs BW (price and resolution factors) 2) resolution 3) sensitivity to low intensity signals (fluorescent applications?) 4) and therefore cooled vs not 5) dynamic range (8 - 12bit) 6) software These are not all but I think the major ones and they are all interelated. It depends on the applications the camera will be used and will it need to be pushed to its specs. Photometrics (on their website) and Princeton Instruments (catalogue) have most of the information in easily understandable language to help you make a decision. ( I have no commercial interests in these companies.) If you want to contact me and I cangive you my reasons for our recent decision on a CCD. Hank Adams EML New Mexico State U. 505-6463600
Hi Robert, About a year or so ago we purschased a RMC tissue processor and have used it pretty much on a daily basis. The main reasons for the RMC over the Leica were: 1) price and 2) I liked the reagent tube caps for each tube, and not the rubber ring over all the tubes. Have had a few minor problems with it, and RMC has been very helpful with repairs.
Any more questions, please feel free to contact me.
Ed Calomeni Medical College of Ohio Dept. Pathology Toledo, OH 43699 emlab-at-opus.mco.edu 419-381-3484
Many thanks for the great response to my request for Bio TEM specimens. Several parties have indicated that they will send a few over. These should more than meet our need.
LocTite 460 is a superior super glue for bonding to all materials. It doesn't bubble and the mechanical strength is the best compared to any other LocTite product available. This is according to what they have told me and my customers are very happy with it.
Bug and tar remover at Chief Auto Parts works very well for removing and dissolving wax from stubs if acetone is not satisfactory. Perhaps a new wax is in order also.
Yours Truly,
Gary Liechty Product Application Specialist Allied High Tech Products, Inc. 2376 E. Pacifica Place Rancho Dominguez, Ca. 90220
please subscribe me to the microscopy listserver Dr. Vickie A. Kimler Assistant Professor of Biology and Allied health Mercyhurst College Erie, PA 16546
e mail: vkimler-at-paradise.mercy.edu Vickie A. Kimler, Ph.D. Biology and Allied Health Department Mercyhurst College Erie, PA 16546 1-814-824-2169
Does anyone have or know of an SEM with cathodoluminescence imginging avaliable for service work (hourly or daily)? I do not need a cold stage, but energy filtering is needed. East coast location would be prefered, southest ideal.
Thanks for any leads or ideas, Phil Russell
Phillip E. Russell Analytical Instrumentation Facility Box 7531, Room 318 EGRC North Carolina State University Raleigh, NC 27695-7531
As this mail list's token optical engineer I guess I should put in a few good words. The fact is that there are many different types of laser confocal microscopes, some of which are direct view, some use a detector and a video monitor, and some are fluorescence. Since the original poster did not specify what microscope she was worried about we have to assume that it is a direct view machine. These use spinning disks (Nipkow disks) or equivalent to raster scan.
Two points should be made: first, and least interesting, is that the manufacturer knows what it is doing and safety of looking into the microscope was worked out at its end and concerns could easily be handled by a call to them.
Second, remember that the laser itself is classified based on the output beam. When that beam is modified by the microscope optics it will have less power spread over a much larger area. Thus the brightness of the beam can be reduced many orders of magnitude before it hits the eye.
It is not intrinsically dangerous to look into a laser. The danger depends on the brightness of the beam. A few months ago a poster on the optics news group reported hooking up deflection and intensity modulators to a video signal and watching TV by writing directly onto his retina. Not something that I would be interested in doing myself, but afterwards he could see good enough to at least type his message.
best regards mark
Mark W. Lund, PhD Director } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"The state is good at simple tasks, like killing people and seizing their wealth. It has far more trouble reaching inside individuals and making them good." Doug Bandow
My laboratory has not used 35 mm film for 12 months. We purchased the SONY DKC5000 camera and have it connected by SCSII drive to a MAC computer. Works great, is very easy to handle and even though the photos do not look so hot when first captured, by using Adobe Photoshop 4.0, and the levels adjustment, within 2 minutes the photo is outstanding. We are printing to a Kodak DS 8650PS printer that has high resolution.
If you would like to view a plate containing 19 photos that never saw film go to the following URL using Netscape:
__________________________________________ Rex A. Hess, Ph.D., Associate Professor, Director, Center for Microscopy and Imaging University of Illinois, Veterinary Biosciences, 2001 S. Lincoln, Urbana, IL 61802-6199 217-333-8933; FAX 217-244-1652; email: r-hess-at-uiuc.edu homepage-- http://www.cvm.uiuc.edu/HomePages/rhess/hesspage.html Center for Microscopy & Imaging -- http://www.cvm.uiuc.edu/MicImagLab/MicImagLab.html Society of Andrology-- http://godot.urol.uic.edu/~androlog/
I'm real new to wedge thinning but as of this morning, I'm having reasonable luck with Loctite's general purpose instant adhesive. It comes in an applicator that actually looks like it may last as long as the glue supply. It's called a SuperBonder Gluematic Pen. Got it at my favorite toy store, W.W.Grainger (www.grainger.com). About $2.50 list.
No interests (other than as a consumer) in either company
cheers, John heckman-at-pilot.msu.edu } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Folks, I have become very concerned that all our talk on pH and Pbcitrate will cause some people to go to their pH meters with their lead solutions to find out the pH. Please do not do this! It almost certainly is not good for the electrode. You will not get an accurate reading anyway, since the pH is will be so high, and the solution is not buffered. What might happen is that you will assess erroneous information from your pH meter to be correct and become embroiled in a vicious cycle of repeatedly making up lead stain attempting to get a correct pH reading. If you follow Reynolds directions, you will automatically get a correct pH. (That is, if you get a true 1N solution of NaOH either from pellets, or from a commercially titrated product). Next week, I will address this problem one more time, and then we all need to go on to something else. Bye, Hildy
Dear Folks, I have become very concerned that all our talk on pH and Pbcitrate will cause some people to go to their pH meters with their lead solutions to find out the pH. Please do not do this! It almost certainly is not good for the electrode. You will not get an accurate reading anyway, since the pH is will be so high, and the solution is not buffered. What might happen is that you will assess erroneous information from your pH meter to be correct and become embroiled in a vicious cycle of repeatedly making up lead stain attempting to get a correct pH reading. If you follow Reynolds directions, you will automatically get a correct pH. (That is, if you get a true 1N solution of NaOH either from pellets, or from a commercially titrated product). Next week, I will address this problem one more time, and then we all need to go on to something else. Bye, Hildy
Allan Mitchell asks why some protocols call for sodium maleate buffers for enbloc staining. There are several reasons: 1. Phosphate and Cacodylate buffers used just previously to UA enbloc have been known to precipitate the UA 2. Rinsing tissue (washing) with maleate buffer, pH 5.2, prevents precipitation of UA in the tissue, and it prevents the minor (but perhaps important) movement of osmium in or out of the tissue. 3. Mixing maleate buffer pH 6.0 with a 3% UA solution 1:1, brings the pH of the UA solution to approximately pH 5.2. 4. This is a refinement, not a necessity, for the achievement of the goal of superior preservation and beautiful micrographs. I have used both concepts to a large extent (no maleate, or maleates) depending on the results I needed. It is more to do if you use maleates. But, if the decision is made to use maleates, they can be made in huge quantities and kept in a freezer. It is called having fun with EM. All the above appeared in detail in the EMSA journal about 10 years ago. It also came up in a workshop given at EMSA by Janet Boyne previous to the journal article. Perhaps it can be located by those interested. Bye, Hily
We will be purchasing a confocal microscope in the near future. It's purpose will be for biological research (no material sciences) and I was wondering if anybody had any strong feelings (positive or negative) about the type of confocal system that they were using, the service from the seller, etc.
We will be purchasing a confocal microscope in the near future. It's purpose will be for biological research (no material sciences) and I was wondering if anybody had any strong feelings (positive or negative) about the type of confocal system that they were using, the service from the seller, etc.
Allan Mitchell asks why some protocols call for sodium maleate buffers for enbloc staining. There are several reasons: 1. Phosphate and Cacodylate buffers used just previously to UA enbloc have been known to precipitate the UA 2. Rinsing tissue (washing) with maleate buffer, pH 5.2, prevents precipitation of UA in the tissue, and it prevents the minor (but perhaps important) movement of osmium in or out of the tissue. 3. Mixing maleate buffer pH 6.0 with a 3% UA solution 1:1, brings the pH of the UA solution to approximately pH 5.2. 4. This is a refinement, not a necessity, for the achievement of the goal of superior preservation and beautiful micrographs. I have used both concepts to a large extent (no maleate, or maleates) depending on the results I needed. It is more to do if you use maleates. But, if the decision is made to use maleates, they can be made in huge quantities and kept in a freezer. It is called having fun with EM. All the above appeared in detail in the EMSA journal about 10 years ago. It also came up in a workshop given at EMSA by Janet Boyne previous to the journal article. Perhaps it can be located by those interested. Bye, Hily
In my experience, most of the molecules in a conventional SEM chamber under normal operating conditions are attributable to water and air, less so to ethanol and isopropanol followed by diffusion and roughing pump oils. Monitoring by residual gas analyzer (RGA) reveals all but the air can be significantly reduced by maintaining the liquid nitrogen baffle above the diff pump. Never open the gate valve to the chamber without the liquid nitrogen trap filled and routinely let the trap warm up with the gate valve closed. Regular maintenance of the molecular sieve trap on the roughing line is also critical.
I hope the EDS manufacturer refered to responds because I don't believe the heater warms the snout or window such that oil would be evolved but rather that is heats the chip to drive ice off the chip face and into the bowels of the dewar. If it does drive off the oil do you really want that oil all over the inside of your SEM? Seems like the solvent trickle method might be best - I have never done it has anyone else had experience with this tricky procedure. Check with your EDS manufacturer before trying it.
Scott Wight
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------------------------------ Scott Wight e-mail: SCOTT.WIGHT-at-NIST.GOV NIST - Microanalysis Group W voice: 301-975-3949 Bld 222, Rm A113 | fax:301-216-1134/301-417-1321 Gaithersburg, MD 20899 \|/ disclaimer: Any opinion expressed is my own and does not represent those of my employer.
Allan Mitchell asks why some protocols call for sodium maleate buffers for enbloc staining. There are several reasons: 1. Phosphate and Cacodylate buffers used just previously to UA enbloc have been known to precipitate the UA 2. Rinsing tissue (washing) with maleate buffer, pH 5.2, prevents precipitation of UA in the tissue, and it prevents the minor (but perhaps important) movement of osmium in or out of the tissue. 3. Mixing maleate buffer pH 6.0 with a 3% UA solution 1:1, brings the pH of the UA solution to approximately pH 5.2. 4. This is a refinement, not a necessity, for the achievement of the goal of superior preservation and beautiful micrographs. I have used both concepts to a large extent (no maleate, or maleates) depending on the results I needed. It is more to do if you use maleates. But, if the decision is made to use maleates, they can be made in huge quantities and kept in a freezer. It is called having fun with EM. All the above appeared in detail in the EMSA journal about 10 years ago. It also came up in a workshop given at EMSA by Janet Boyne previous to the journal article. Perhaps it can be located by those interested. Bye, Hily
Mark, Plastic Pages-4 pocket for 3 ring binder are available From IMBROS in Moonach, Tasmania Tel 03 6273 1300 If you wish we can supply them from the USA. Ladd CAT# 11-81250
Preece, T. 1978. Toluidine blue: The staining method of Shoremaker..... FBPP News 1:40-40.
Can anyone tell me what "FBPP" stand for? (note: is apparently is not "Federation of British Plant Pathology")
TIA
Robert R. Wise Plant Physiologist and Director, UWO Electron Microscope Facility Department of Biology University of Wisconsin Oshkosh Oshkosh, WI 54901 (414) 424-3404 tel (414) 424-1101 fax wise-at-uwosh.edu
I am interested in procuring a cryostage that is in good working condition for our ZEISS 902 Energy Filtering TEM. If anybody who has one and wants to part with it can contact me directly. Thanks!
M.V.P.
************************************************************************* M.V. Parthasarathy Professor of Plant Biology & Director, Cornell Integrated Microscopy Center (CIMC) Section of Plant Biology 228 Plant Science Building Cornell University, Ithaca, NY 14853 Telephone: Plant Biology Office (607) 255-1734 Fax: " (607) 255-5407 Telephone: CIMC Office (607) 253-3803 Fax: " (6)7) 253-3803 E-mail: mvp2-at-cornell.edu ***********************************************************************
I am interested in procuring a cryostage that is in good working condition for our ZEISS 902 Energy Filtering TEM. If anybody who has one and wants to part with it can contact me directly. Thanks!
M.V.P.
************************************************************************* M.V. Parthasarathy Professor of Plant Biology & Director, Cornell Integrated Microscopy Center (CIMC) Section of Plant Biology 228 Plant Science Building Cornell University, Ithaca, NY 14853 Telephone: Plant Biology Office (607) 255-1734 Fax: " (607) 255-5407 Telephone: CIMC Office (607) 253-3803 Fax: " (6)7) 253-3803 E-mail: mvp2-at-cornell.edu ***********************************************************************
The thread on solvent cleaning of stubs has turned into a solid rope, perhaps we can twist it into a 4 inch hawser. I like Bill Chissoe III's variation: Brute force and not those nasty solvents. A medium-fine flat file, lying on a bit of paper towel served me well to clean the residues from the top of stubs. A suede wire brush is effective for occasional cleaning of the file. I believe that method its less messy and often faster than those solvents.
Economics, however, also come into such considerations. Every now and then somebody rediscovers that by going back in the literature they can find the methods to refilament, make thin film apertures, various standards or even LaB6 and they could make them for the lab. One in a thousand may have good reasons to do so.
Generally, cleaning up stubs is still worthwhile. A stub costs A$0.32 or about US$0.25. Amazingly some labs even manufacture new stubs without a production lathe "in house". An employer has to count "on cost" and the cost of the facilities. Commercially it makes sense to "recover" triple the hourly pay for any task before break-even point is reached. A person on $15/h needs to clean up 180 stubs/h to break even so far as the institution is concerned. Clearly, Professors should work faster or delegate hum drum chores.
Some thirty years ago the most commonly used Cu grids cost A$7.00/100. They now cost A$10 and although our currency has depreciated, it is obvious that grids are much cheaper now in relative terms. I recycled thousands of grids up to about 25 years ago. Today this would make no sense whatever - unless you are working in a cheap labour country.
Of course, we must not forget institutional economics which work to different realities: "We only have so much (too little) money for maintenance; we're paying that person anyway. Productivity? Who cares, we're only trying to balance our budget."
Stop reading your email now and do something, CLEAN THOSE STUBS! Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 350+ Links, MSDS ************************ http://www.proscitech.com.au
} } If I am off-base with this response, I apologize. I have used nothing } but carbon tape ever since it became readily available through regular } EM suppliers but I always dreaded having to clean it off my sample } holders because it was so sticky and tore so easily. Being a simple } minded person, I just took a 3/8" wooden dowel, sharpened one end(like a } chisel) and used it to roll the tape up into a wad that I could remove } with my fingers. All the adhesive seems to comes off with the tape, but } if any should be left behind it should easily come off with a mild } solvent. (I always polish my holders, so I'm not sure about residual } adhesive.) I also cut a piece of latex tubing to fit on the end of the } rod to cushion my hand. This works for the carbon tape, I'm not sure } about the other types of adhesive tapes. For what it's worth. } } Bill } -- } ============================================================= } Bill Chissoe III } Electron Microscopist,University of Oklahoma } E-mail: wchiss-at-ou.edu Ph. (405)325-4391 } =============================================================
The Microbeam Analysis Sciety is pleased to announce the introduction of a new mailing list (listserver) dedicated to electron microprobe analysis. This mailing list is sponsored by MAS and is managed by two MAS Directors, John Mansfield and Greg Meeker. It is being formed in response to several requests from members of MAS. The list is open to anyone interested in microprobe analysis. To subscribe you may send mail to the mailing list at:
microprobe-at-www.microanalysis.org
with the word "subscribe" in the subject.
You may also subscribe by connecting to:
http://www.microanalysis.org/mas/maslistserver/maslistserver.html (there are also links from the MAS home page http://www.microanalysis.org/mas
A message will be returned telling you that you are subscribed to the list and listing the commands that may be sent to the mailing list. A copy of this message file is included below.
Once you are subscribed you may send messages to the list by sending mail to:
microprobe-at-www.microanalysis.org.
Comments, Criticisms (mild ones only please! :-) ) to:
John Mansfield & Greg Meeker jfmjfm-at-umich.edu and gmeeker-at-usgsprobe.cr.usgs.gov respectively.
________________________________ Copy of the subscribe welcome message follows:
Welcome, you are now SUBSCRIBED to the Microprobe Listserver, a mailing list for users of wavelength dispersive X-ray analysis systems.
You are subscribed in INDIVIDUAL MESSAGE mode. Each time a message is sent to the mailing list, a copy will be forwarded to you. If you want to set your messages to digest mode check the "Help Section" below for instructions.
This list is sponsored by The Microbeam Analysis Society (http://www.microanalysis.org). This list is unmoderated, but you must be subscribed to send and receive mail to and from the list.
This list is overseen by John Mansfield and Greg Meeker. Questions about this list can be directed to either person, however, you should check the "Help Section" below to see if that can answer your questions before contacting them. We should also note that Greg is an active microprobe user, whereas John is not, technical questions about microprobe analysis should, therefore, be addressed to Greg. Greg and John can be contacted as follows:
Greg Meeker U.S. Geological Survey Mail Stop 903 Box 25046, DFC Denver CO 80225 Phone: (303)236-1081 Email: gmeeker-at-usgsprobe.cr.usgs.gov
John F. Mansfield North Campus Electron Microbeam Analysis Laboratory University of Michigan 417 Space Research Building 2455 Hayward Ann Arbor MI 48109-2143 Phone: (313)936-3352 or (313) 715-2510 FAX (313)936-3352 or (313) 763-5567 Email: jfmjfm-at-engin.umich.edu or John.F.Mansfield-at-umich.edu URL:http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html
Help Section
If you have colleagues who wish to join this list, remember this listserver supports the following commands. These commands should be the text in the subject field of your mail message addressed to: microprobe-at-www.microanalysis.org.
subscribe Your address will be added to the list of subscribers. You will then be able to send messages to this list that will be forwarded to all other list subscribers.
subscribe digest Your address will be added to the list of subscribers who receive a digest instead of each forwarded message. You will be able to send messages to this list, and will receive a digest of accumulated messages once a day.
digests A list of available digests will be returned to you.
unsubscribe Your address will be removed from the list of subscribers. You will no longer be able to send messages to the members of the list.
help This help section will be returned.
End of subscription message.
John Mansfield North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (313)936-3352 FAX (313)936-3352 Email: jfmjfm-at-umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html
I want to be able to grab S-VHS video in my camputer for sending through the web. I have a video camera mounted on my trinocular Nikon and save as S-VHS on VCR'S. I have heard of an add-on called SNAPPY. Has anyone used this and does it give good images? Can anyone supply names of video cards?
Mike Dingley -- Check out the P.M.C.A. Homepage and Mike's Freshwater Algae Homepage at http://www.pnc.com.au/~dingley
Is anyone aware of a solvent which dissolves polymerized LR White? I have two pieces of aluminum, separated by a small gap, which infiltrated with LR White prior to UV polymerization. Now they are hopelessly stuck together. I've tried methylene chloride with no success, and am now searching for other ideas. I can work in the hood, and considering the importance and value of these parts, I am willing to take what ever precautions a particular solvent might require. Acids which might attack aluminum are unfortunately not an option. Any ideas?
Many thanks,
Doug Keene Shriners Hospital for Children Portland, Oregon ---------------------- Doug Keene DRK-at-shcc.org
N,N,dimethylformamide might be worth a whirl, I think I used to use it years ago to dissolve cured epoxy resins for subsequent chemical analysis. I don't know about LR White, but DMF dissolves lots of things.
cheers
Ritchie
} Is anyone aware of a solvent which dissolves polymerized LR } White? I have two pieces of aluminum, separated by a small } gap, which infiltrated with LR White prior to UV } polymerization. Now they are hopelessly stuck } together. I've tried methylene chloride with no success, } and am now searching for other ideas. I can work in the } hood, and considering the importance and value of these } parts, I am willing to take what ever precautions a } particular solvent might require. Acids which might attack } aluminum are unfortunately not an option. Any ideas? }
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
I would be grateful for information on daily/weekly workloads expected = of technical staff in diagnostic TEM labs (pathology). For example: = number of specimens received and processed (by hand or automatic = processor), sections cut, stained and viewed, photographics (including = printing), other duties included, etc. Also, I am interested in = determining what is considered an efficient diagnostic TEM pathology = service in terms of numbers of specimens processed per year against = total staffing levels (all staff) and specimen turn-around times. If you do not want your information to be seen on the MSA listserver, = mail can be sent direct to: John.Stirling-at-flinders.edu.au
Thanks in advance! John W Stirling Department of Anatomical Pathology Flinders Medical Centre Bedford Park SA 5042 Australia=20
I think that a cup of boiling water may help you to separate the samples. The epoxy might not be dissolved in the hot water, but it would become soft enough to let you tear the aluminum off.
With the best wishes,
Charlie Kong EM Unit, UNSW, Australia
On Mon, 5 May 1997, Doug Keene wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Is anyone aware of a solvent which dissolves polymerized LR } White? I have two pieces of aluminum, separated by a small } gap, which infiltrated with LR White prior to UV } polymerization. Now they are hopelessly stuck } together. I've tried methylene chloride with no success, } and am now searching for other ideas. I can work in the } hood, and considering the importance and value of these } parts, I am willing to take what ever precautions a } particular solvent might require. Acids which might attack } aluminum are unfortunately not an option. Any ideas? } } Many thanks, } } Doug Keene } Shriners Hospital for Children } Portland, Oregon } ---------------------- } Doug Keene } DRK-at-shcc.org } }
Edward J. Basgall wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } } ------- Forwarded Message Follows ------- } } } Does anyone know the dangers associated with a technique called Confocal } } Microscopy or something like that? In our Chemistry Dept. we have a } } research group that works with a Class 4 LASER. To the best of my } } understanding, the LASER beam is directed toward a sample that is sitting } } under a microscope. Some of the beam is reflected and filtered so the } } power or strength of the beam is reduced before it hits the sample } } (a biological sample). The researcher then looks through the microscope } } objective at the sample. } } } } Currently, we have a LASER safety program in place. This particular } } procedure concerns me because this technique allows the researcher to } } look directly at the LASER beam. This does not sound too good. Also, } } the microscope objective may magnify the beam and direct the beam } } toward the researchers eye. } } } } Currently, the LASER that is being used is an Class 4 ARGON Ion LASER } } (Continuous Wave) with a wavelength of 0.488 um (ultraviolet range) and a } } beam power of 1.5 W. } } } } Does anyone know where I can get more information on this subject. I am } } particularly interested in the safety aspects of this technique. It } } may be the case that the LASER beam hitting the sample has a low enough beam } } power to classify it as a Class 1 LASER. } } } } Thanks in advance for your help. } } } } Laurie Princiotto } } Laboratory Safety Specialist } } Indiana University - Dept. of Environmental Health and Safety } } Phone: (812)-855-6115 } } Fax: (812)-855-7906 } } E-mail: lprincio-at-indiana.edu } } Laurie, } This does not sound like a very safe activity to me. I would be very } concerned until proven safe. } } Regarding the lasers used in confocal microscopes...I've worked with the } Bio Rad commercial system and I believe most use a class II Ar or Ar/Kr } laser at ~1mW power (457-648nm). The laser image cannot be directly } observed but is displayed in a computer screen. } } You're best info sources would be the manufacturers of confocal microscopes. } } Leica - Ph: 415-348-2233 } Bio-Rad - Ph: 800-4BIORAD } Molecular Dynamics - http://www.mdyn.com ph:800-333-5703 or 408-773-1222 } Zeiss 800-233-2343 } } good luck } } Edward J. Basgall, PhD } The Pennsylvania State University } Surface Chemistry Group ejb11-at-psu.edu } Materials Research Institute Building Ph: 814-865-0493 } University Park, PA 16802-7003 FAX: 814-863-0618 Dear Ed et. al.,
As past technical marketing manager for Sarastro (now Molecular Dynamics), I can assure you that when these microscopes are designed, they must meet extremely stringent safety standards. One of two options is used: either existing optics are chosen which eliminate any possibility of laser light reaching your eyes (ex: when we used a Nikon microscope as the basis for the Sarastro system, we used an "F-head" in which the full binocular body rotated out of position when the laser was directed to the sample) or optics are designed with fool-proof safety mechanisms. One of the previous respondents had the imaging story partially correct: In confocal microscopy, the laser scans the sample very much like a TV raster scan. However, the light goes to a detector (either a photomultiplier tube or a video camera). From that point, the signal passes to a computer for processing and imaging. There are some "direct view" confocal systems, notably, those produced by Technical Instruments (San Francisco), but these are not laser systems . Instead, they use the same sort of "white light" illuminator you would normally find on a regular light microscope. Hope this helps.
} Jim: } I'd be very interested to know what method you would recommend for } cleaning Be windows on EDS detectors (and probably so also would a lot of } others on the Microscopy Listserver). } } We have done it several times in the past by using a dropper to run a } bit of petroleum ether over the face of the window. You have, of course, } to be very careful not to puncture the window with the dropper, but then } the whole operation is one of high risk to begin with. Pet ether is } basically a low-boiling hydrocarbobn fraction that is a pretty good solvent } for other hydrocarbons, but which does not attack epoxies and which also } evaporates quite readily and cleanly. We have even been able to do this } without removing the detector from the SEM by using a long dropper and } collecting th Pet ether on a tissue as it runs off the detector. I am not } sure how well this might work with silicone oils, because of their limited } solubilities; however, one old-time industrial solvent for them was } kerosene, which is again basically a hydrocarbon material. } } Wilbur C. Bigelow, Prof. Emeritus } Materials Sci. & Engr., University of Michigan } Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; } Fx:313-763-4788; Ph:313-764-3321 *********************************** Wil: Your method was fairly similar to ours. You and I would not use silicone in an EM, but if kerosene was used, a second solvent would be needed to remove the kerosene residues. Because I worked for many years in a rather isolated lab (1500km to the nearest other electron beam instrument) factory maintenance was never an option and for a time we had three EDS. Link suggested that reagent grade methanol was a most suitable solvent. Presumably this was recommended because it did not affect the Be window's glue and it is a good solvent of vacuum oils and fluids. Happily we too could leave the detector in place and I collected the used solvent in a Petrie dish and also with tissue. The significant difference in our methods was the application. Because the window is recessed and our detector had some tilt, it seemed that little solvent would reach the window if the methanol was just dripped from the top.
Use polyethylene Pasteur type pipette with a fairly large bulb. Pull the tip into a fine capillary after brief (5-10 seconds) heating in a low flame. (Tip: move and rotate pipette while heating; pull vertically, this keeps the tip straight while the plastic sets) Cut the tip so that about 100mm of capillary tip and a small bore, perhaps 0.2mm is available. FILL the bulb with methanol; releasing air once or twice by up-turning and squeezing the bulb. Filling is slow but no trouble. Freehand application is possible with care and a steady hand, but most people would be happier mounting the pipette horizontally in a low "retort" stand, at the Be window's level. Slide the stand until the pipette tip is within 20 to 40mm of the window. Hold pipette with one hand (hand should rest, never work mid-air) near the tip and gently squeeze the bulb. The aim is to "hit" with little force the upper rim of the Be window - where is supported. Less force on the window and also slightly better coverage is achieved with the methanol stream directed well away from the perpendicular.
A side benefit of well tilted detectors is that cleaning is much easier: Simply use a suitable vial half filled with solvent and slide snout into this. Slightly swirl and cleaning is done. Regards Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 350+ Links, MSDS ************************ http://www.proscitech.com.au
Hello all; Is anyone out there doing in situ hybridization using the DIG kit from Boehringer-Mannheim? We have been trying to use the kit, and have been having trouble with a couple of things....Has anyone out there ever gotten the blocking agent to dissolve at 0.5% (every time I try, I get what looks like poached egg white, regardless of how gently or vigorously I heat it!) I can't remember who sent the suggestion to someone else to try cold water fish skin gelatin for blocking, but bless you!! It seems to be of some help. Is it possible to "overdetect" with this system? I keep thinking I've overdone it, but when I get the slides dehydrated and Permounted, things aren't very dark (not like what I've seen in the literature) In the DIG applications manual from B-M, I notice that if you're doing blots, the colour is different depending on the membrane used (brownish on nylon, blueish on nitrocellulose). What types of colours are people getting on what types of tissues (I'm looking at soybeans, by the way). Sorry if this sounds incoherent...it is, after all, Monday morning.
Thanks in advance shea
Dr. S. Shea Miller Agriculture & Agri-Food Canada Eastern Cereal & Oilseed Research Centre Rm 2068, Bldg 20, CEF Ottawa, Ontario Canada K1A 0C6 Phone: (613)759-1760 Fax: (613)759-1701 e-mail: millers-at-em.agr.ca
At Advanced Imaging Concepts we make software for archiving and databasing digital images and also handle a wide variety of frame grabbers and systems. The Snappy you mentioned is a parallel port device that only handles composite video, not Y/C. Please contact me and I will be glad to discuss a number of options with you.
Scott E. Berman Advanced Imaging Concepts, Inc. Princeton, NJ Phone(609) 921-3629 Fax(609) 924-3010 email Scott E57-at-aol.com
There is a discussion on this archived at "Tips & Tricks" Go to the web address at the end of this message and click on "Tips & Tricks. In the "TEM" section you will find a link to "Dissolving Methacrylates" which has what you need. Good luck.
At 10:59 PM 5/5/97 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 352-392-1295 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
I felt that this would be a good place to ask this question and hopefully I will get lots of good advice:
I want to know if anyone knows where and/or how I can puchase 2.5% calcium gluconate gel.
I have purchased hydroflouric acid from Aldrich Chemical Co. and in their MSDS sheet they recomend this gel as a neutralizing agent if you should happen to get any HF on your skin. I am quite aware of how extremely dangerous HF is and I do not want to use it for any of my etching experiments until I have all my safety precautions taken care of.
I called Aldrich and they do not sell the calcium gluconate and they were of absolutely no help in letting me know where I could find it and they did not give me any readily available alternatives.
Does anybody have any clues or suggestions as to where I can get this calcium gluconate gel or for that matter, do you have any helpful suggestions about dealing with this extremely nasty chemical. If I had my choice I would not use it at all, but I must.
Thank you for your help.
Margaret E. Bisher
NEC Research Institute 4 Independence Way Princeton, NJ 08540. Tel.: (609) 951-2629 Fax: (609) 951-2496 e-mail: peggy-at-research.nj.nec.com
I've had some expierience with several confocal systems. Our lab currently has an InSignt and Ultima form Meridian Instruments. The Ultima is more specialized for cytometry, does a wonderful job, but requires a full-time supervisor. The InSight is brand new and looks promising. It has not been used enough to make any judgments.
I personally use an older Zeiss LSM (model 20, I believe) and a Biorad MRC600. Both provide good general purpose use. I would suspect the upgrades of these machines would be very useful. (The limits on these systems for me is that they do not allow ratiometric imaging--that is the older models)
All of this is IMO, alone and I have no financial interest in any of these companies.
Good luck.
Tommy C. Sewall Image Analysis Laboratory College of Veterinary Medicine Texas A&M University
} I want to know if anyone knows where and/or how I can puchase 2.5% calcium } gluconate gel. } The three readily available chemical catalogs--Alfa Aesar, Aldrich and Sigma--all have gluconates, and Aldrich and Sigma have Ca-salts (you could, of course, add CaOH to the acid if need be). I guess that adding the right amount of H2O will give the 2.5% gel.
} I have purchased hydroflouric acid from Aldrich Chemical Co. and in their } MSDS sheet they recomend this gel as a neutralizing agent if you should } happen to get any HF on your skin. I am quite aware of how extremely } dangerous HF is and I do not want to use it for any of my etching } experiments until I have all my safety precautions taken care of. } } I called Aldrich and they do not sell the calcium gluconate and they were } of absolutely no help in letting me know where I could find it and they } did not give me any readily available alternatives.
Obviously, their catalog and sales reps are not on the same page! } } Does anybody have any clues or suggestions as to where I can get this } calcium gluconate gel or for that matter, do you have any helpful } suggestions about dealing with this extremely nasty chemical. If I had my } choice I would not use it at all, but I must. } Use it in a hood, wear a lab coat and polyethylene gloves, and use goggles or a face shield. H2F2 is an inhalation hazard, so make sure your hood has adequate air flow. H2F2 is nasty, but not as nasty as some of the chemicals in use in organic chem labs. Good luck. Yours, Bill Tivol
OK, so Tilex was on sale this weekend, and I bought some. Never mind that when I once used it in my shower years ago I was mightily offended by the smell. Let's put together a list of all the wonderful uses this group has come up with! I'm sure the Clorox Company would be amazed. This sounds like the duct tape of the solvent world. Contains diethylene glycol butyl ether.
*Duct tape is like the Force; it has a dark side and a light side and holds the Universe together.*
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
You may also want to check with manufacturers of such equipment. We maintain a database of sample preparation methods and can often give references fr specific preparation needs. We also publish a bibliography of technical papers dealing with sample preparation which we are pleased to send you at no charge. You can then select papers of interest and we will send you reprints at no charge.
David Henriks TEL: 800-728-2233 (toll-free in USA) South Bay Technology, Inc. 714-492-2600 1120 Via Callejon FAX: 714-492-1499 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 Juan Marti } Hello EVERYBODY,
Does anyone out there knows if there is a specific list about Surface preparation methods for LM (Grinding, Polishing, etching...etc)?
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Materials technician positions are available immediately to qualified persons as SEM and TEM operators in the Process Characterization Analysis Laboratory at AMD's Austin manufacturing facilities. The positions require demonstrated experience in the preparation of samples associated with the microelectronics industry as well as basic understanding of materials involved in submicron CMOS technology. Good organizational skills are a must. Good interpersonal and communication skills in a multidisciplinary environment are expected. Additional skills in the operation of state of the art TEM or SEM's would be helpful. These are career path positions and allow contribution individually or in teams in a cooperative environment. An Associate degree in electron microscopy, materials science or a physical science is needed. Directly related experience will be considered in lieu of a degree. Send resumes to David Valdez, AMD, 5204 E. Ben White Blvd., M/S 556. Austin, TX - 78741; FAX (512) 602-5108. AMD is an equal opportunity employer.
Mr-Received: by mta RANDD; Relayed; Mon, 05 May 1997 14:25:12 -0500 Mr-Received: by mta MCM$RAND; Relayed; Mon, 05 May 1997 14:25:13 -0500 Mr-Received: by mta RANDB; Relayed; Mon, 05 May 1997 14:25:20 -0500 Alternate-Recipient: prohibited Disclose-Recipients: prohibited Content-Return: prohibited
About three years ago, I spent several months trying to use the Boehringer digoxygenin system on lung tissue sections. The blocking agent was a huge pain to get into solution. I had several conversations with the technical reps about this - and got conflicting recommendations on how to get it in solution. One of the methods was to heat the solution in a microwave processor - the result was a carmelized goopy mess. I was also told to heat water to 68C on a hot plate and add the blocking solution powder while stirring vigorously; the key, I was told, was to avoid a rolling boil. That worked fairly well - until I autoclaved it. When I pulled my containers out of the autoclave, I found something that resembled soupy cottage cheese. As I recall, the next time I made the solution, I used the hot plate method and ignored Boehringer's recommendation to autoclave it. I eventually switched to P-33 and autoradiography, as the mRNA I was looking for was not abundant, so I never became an expert at using the digoxygenin method.
About the color reaction: if you are using BCIP/INT as the chromogen for alkaline phosphatase (as I remember, that's what Boehringer gives you), the reaction product is soluble in organic solvents. You may be losing it if you are mounting your slides in Permount. There are aqueous mounting media that will work better for this chromogen.
Good luck!
Jane A. Fagerland, Ph.D. Dept. Microscopy and Microanalysis Abbott Laboratories Abbott Park IL 60064
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear MSA Newsgroup:
I felt that this would be a good place to ask this question and hopefully I will get lots of good advice:
I want to know if anyone knows where and/or how I can puchase 2.5% calcium gluconate gel.
I have purchased hydroflouric acid from Aldrich Chemical Co. and in their MSDS sheet they recomend this gel as a neutralizing agent if you should happen to get any HF on your skin. I am quite aware of how extremely dangerous HF is and I do not want to use it for any of my etching experiments until I have all my safety precautions taken care of.
I called Aldrich and they do not sell the calcium gluconate and they were of absolutely no help in letting me know where I could find it and they did not give me any readily available alternatives.
Does anybody have any clues or suggestions as to where I can get this calcium gluconate gel or for that matter, do you have any helpful suggestions about dealing with this extremely nasty chemical. If I had my choice I would not use it at all, but I must.
Thank you for your help.
Margaret E. Bisher
NEC Research Institute 4 Independence Way Princeton, NJ 08540. Tel.: (609) 951-2629 Fax: (609) 951-2496 e-mail: peggy-at-research.nj.nec.com
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Materials technician positions are available immediately to qualified persons as SEM and TEM operators in the Process Characterization Analysis Laboratory at AMD's Austin manufacturing facilities. The positions require demonstrated experience in the preparation of samples associated with the microelectronics industry as well as basic understanding of materials involved in submicron CMOS technology. Good organizational skills are a must. Good interpersonal and communication skills in a multidisciplinary environment are expected. Additional skills in the operation of state of the art TEM or SEM's would be helpful. These are career path positions and allow contribution individually or in teams in a cooperative environment. An Associate degree in electron microscopy, materials science or a physical science is needed. Directly related experience will be considered in lieu of a degree. Send resumes to David Valdez, AMD, 5204 E. Ben White Blvd., M/S 556. Austin, TX - 78741; FAX (512) 602-5108. AMD is an equal opportunity employer.
FESEM Engineer FAB 12 Yield Dept. Materials Lab Intel Corporation
The successful candidate will be a materials analyst in the FAB-12 Materials Lab. Responsibilities will include interacting with process, reliability and materials/LYA lab engineers in resolving materials issues in manufacturing and improving process yields by performing required analysis and providing the team with data interpretation and recommendations. Initial responsibilities will be focused on supporting the Scanning Electron Microscopy (SEM) lab. The candidate will be required to provide direction/supervision for 2-3 technicians in sustaining SEM support. Must have BS or MS in Materials Science, Chemical Engineering or equivalent with work experience or relevant coursework in materials analysis related to the semiconductor industry.
Skills:
Excellent communication/interpersonal skills required. The successful candidate should be able to prioritize multiple tasks to effectively meet customer needs in a changing environment. Must have hands on experience with Scanning Electron Microscopy (SEM).
Intel is an equal opportunity employer. Resumes accepted by fax, email, or snail mail. Interested candidates should contact:
Satish Naidu Manager-F12 Materials Lab Intel Corporation M/S OC2-218 4500 S. Dobson Rd Chandler, AZ 85248-4906 satish_k_naidu-at-ccm.ch.intel.com Fax(602)715-8363
I clean mine by gently dribbling Freon over the window, a gentle stream from a plastic dropper bottle (the sort that has a nozzle as its top and that you squeeze), directing the stream to the metal above the window. I feel a bit bad about using Freon, but it's such a great solvent for grease and oil, and so non-aggressive towards epoxies and things that I justify my continued use of the stocks which I inherited by the absense of any disposal facilities in NZ. When it runs out I think I'll use pet ether.
cheers
Ritchie
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
I do not not if this might help you. I am using HF on a regular basis. Here in Australia I can get the Calcium Gluconate Gel from ORION - Laboratories P/L - Briggs Street - Welshpool - Western Australia.
Hans Brinkies Senior Lecturer SWINBURNE, University of Technology School of Engineering and Science HAWTHORN - Vic. - 3122 - Australia
This reply is mostly a soapbox digression from the microscopy theme of the listserver so DELETE or "click past" now if you're busy. I would suggest sending any replies to me personally rather than cluttering up the list.
To Ritchie and others worried about CFC (freon) use,
Dab away merrily with your freon soaked swab. And do so without guilt. Freon is one of humanity's greatest creations. Inert, non toxic, non-flammible and the planet's most cost effective refrigerant, this compound and other CFCs have gotten a bum wrap. The new CFC-free replacement compounds are corrosive, toxic, expensive and only 75 to 80% as efficient. At least one toxic spill/bird kill has already occurred. My 70's vintage Neslab Cryocool can crank -100 C after 25 years. When it breaks, I can only get the new CFC-free model can achieve no more than -80 C and will not last as long. Absolutely no disrespect to Neslab intended or deserved.
Stratospheric ozone depletion measurements were not taken before the 1970's. "Ozone Depletion" may turn out to be natural ozone variation. We won't know for centuries. O3 to O2 CAN occur via chlorine and fluorine catalytic pathways. That man-made CFCs are the catalysts can never be proved since chlorine and fluorine are abundant natural elements. Maximum reported CFC concentrations in the stratosphere are in the area of 3 parts per trillion (Nasa?). Seems kind of sparse if true. Can anyone tell me if that kind of detection limit is reliable for any instrument?
Maximum increased "plus hundred year" UV fluxes related to projected stratospheric ozone thinning, CFC caused or otherwise, are comparable, in skin cancer health risk to moving a few hundred miles South or a thousand feet up in elevation. Pale northerners taking winter vacations in southern latitudes will suffer radical UV induced cancer risk due to insufficient melanin buildup, regardless of ozone layer variations. (Don't forget to smear the PABA 16 between your toes, the new high risk site for benign skin cancer.) In my understanding malignant melanoma is not directly associated with UV exposure.
The Montreal Protocol back in 89-90 created an agreement among the industrialized nations that CFCs would be banned from manufacture in 1995. E.I. duPont was the principal supplier of CFCs, and yet duPont was an important financial contributor to the physical execution of the meetings for the Montreal Protocol. 1995 was also the year that duPont lost their patent on CFCs. Seem suspicious? I WOULD SAY SO, when you consider that they also hold the new patents on the legal CFC replacements. And... please forgive me or correct me if I am wrong about this, but I have heard in conversation with industrial suppliers that in 1995 duPont fired up a large new CFC manufacturing facility in India to supply refrigerants for the anticipated third world appliance boom. Once again, special interests make the planet go-round.
I have no direct financial stake in CFC issues. I just hate losing another great material and suffering useless regulatory expenses as a result of hasty science and ill conceived politics.
I welcome all elaborations and criticisms on the above. Ozone and asbestos paranoia are my pet peeves. I would only be happier to learn that our sacrifices are not in vain.
We have a new Polaron CPD and are having difficulty maintaining the carbon dioxide level when flushing. A check for leaks, revealed a small leak at the pressure gauge, which new seals fixed. However the level problem still persists. The temperature is kept around 12 degrees celisus. Could this be the inlet valve? Has anyone else had this problem and what did you do to fix it. TIA
Mary Toogood Atlantic Food & Horticulture Research Centre Agriculture and Agri-Food Canada Kentville N.S. B4N 1J5 ToogoodM-at-em.agr.ca
Thanks for all your good solutions (yuk, yuk) for removing tapes/spots from Al mounts. You have given me just the ammunition I need to justify a $50K (US) supercritical CO2 organic extraction system.
Seriously, I used a commercial paint/label remover, Goof-off and a little elbow grease. It worked fine. I then immersed the stubs in an ultrasonic bath of warm (40C) DI water and 0.5% by volume Micro for about five minutes followed by a DI rinse and air dry. I haven't looked at them in the SEM yet, but at optical mags up to 40x, they appear clean even when viewed using a UV lamp.
Thanks again. (Note to vendors: I still buy new ones.)
------------------------------------------------ Opinions or statements expressed herein, rational or otherwise, do not necessarily reflect those of my employer.
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 Phone: (508) 750-1717 E-mail: crossman-at-osi.sylvania.com
Our web sites: www.sylvania.com www.siemens.com --
How does one go about getting added (and deleted) from your mailing list?
MG
Dr. Michael G. Gorczyca, Ph.D. Department of Biology Morrill Science Center University of Massachusetts Amherst, MA 01003 tel: (413) 545-1783 or -4627 fax: (413) 545-3243 e-mail: mgor-at-bio.umass.edu
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello, I have a Zeiss DSM 960a with an Oxford Link EDS system. The EDS spectra has a large noise peak, typically 10 times the dominant real peak. I have been talking with Oxford about this a great deal and they are unsure of the cause. I now ask you for input.
I checked the electrical isolation between the dewar and the column and found there is none (1 Ohm) if the cables are connected and infinite if they are disconnected. To Oxford's suprise they found the same on their system. Is this normal? This means that noise in the SEM is in the EDS!? How should I be grounding this system. We do have a 1 cm diameter copper wire that is a dedicated ground running up to these to systems. Any general tips on electrical issues for EDS would be appreciated.
thanks
Brad Storey Materials Scientist Argonne National Lab - West P.O. Box 2528 Idaho Falls, ID 83403 Ph. 208-533-7685 Fax 208-533-7683 e-mail brad.storey-at-anl.gov
Message-Id: {199705061459.KAA22195-at-umic.sunysb.edu} Comments: Authenticated sender is {greg-at-mail.umic.sunysb.edu}
Mary, Maintaining the level of CO2 is a balancing act. It must be watched!! As the CO2 drains it freezes at the valve blocking the opening. This can slow draining. Then it will blow free and the rate of draining speeds up. The flushing should be done several times for a few minutes each time rather than continually. Hope this helps.
} } We have a new Polaron CPD and are having difficulty } maintaining the carbon dioxide level when flushing. A } check for leaks, revealed a small leak at the pressure } gauge, which new seals fixed. However the level problem } still persists. The temperature is kept around 12 degrees } celisus. Could this be the inlet valve? Has anyone else } had this problem and what did you do to fix it. TIA } } } } Mary Toogood } Atlantic Food & Horticulture Research Centre } Agriculture and Agri-Food Canada } Kentville N.S. B4N 1J5 } ToogoodM-at-em.agr.ca } Gregory Rudomen Greg-at-UMIC.SUNYSB.EDU 516-444-3126 University Microscopy Imaging Center S.U.N.Y. Stony Brook
Dear all, A week ago, I sent the following message to the list.
} Dear all, } I am aware of a number of programs that can produce calculated diffraction } } patterns given the unit cell parameters and the location of all atoms in } the } unit cell. This poses difficulties for some materials with more } complex } structures, however, where site occupancies are not 100% and } where the 'unit } cell' is somewhat of an average structure (this occurs } for some oxides). This } can be overcome if a good crystal structure } refinement has been done from XRD } where average atomic positions and site } occupancies have been determined. Many } structures have not been studied } in this detail, however, and the only } information that is available is } the unit cell parameters and the space group. } Is there any software that } can plot electron diffraction patterns from this } limited information. } } I hope there is someone who can help me with this.
I have received a number of helpful suggestions. These include using Desktop Microscopist (for Macintosh) from Virtual Labs. We have this program here (version 1.05) but I've had problems running it on my computer. Some other prgrams were mentioned that we don't have and haven't been tried. Phillipe-Andr=E9 Buffat at EPFL in Switzerland recommended the use of EMS online via the WWW at http://cimewww.epfl.ch/ and this has proved very useful and I will probably use it at some point.
Thanks to all who responded
Yours sincerely
Ian MacLaren
P.S. A copy of the responses can be received by emailing me.
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
Dear all, A week ago, I sent the following message to the list.
} Dear all, } I am aware of a number of programs that can produce calculated diffraction } } patterns given the unit cell parameters and the location of all atoms in } the } unit cell. This poses difficulties for some materials with more } complex } structures, however, where site occupancies are not 100% and } where the 'unit } cell' is somewhat of an average structure (this occurs } for some oxides). This } can be overcome if a good crystal structure } refinement has been done from XRD } where average atomic positions and site } occupancies have been determined. Many } structures have not been studied } in this detail, however, and the only } information that is available is } the unit cell parameters and the space group. } Is there any software that } can plot electron diffraction patterns from this } limited information. } } I hope there is someone who can help me with this.
I have received a number of helpful suggestions. These include using Desktop Microscopist (for Macintosh) from Virtual Labs. We have this program here (version 1.05) but I've had problems running it on my computer. Some other prgrams were mentioned that we don't have and haven't been tried. Phillipe-Andr=E9 Buffat at EPFL in Switzerland recommended the use of EMS online via the WWW at http://cimewww.epfl.ch/ and this has proved very useful and I will probably use it at some point.
Thanks to all who responded
Yours sincerely
Ian MacLaren
P.S. A copy of the responses can be received by emailing me.
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
When you say "noise peak", are you referring to the strobe at 0 keV? Ours is generally comparable to the major peaks in our spectra for an Isis 300 system on a Hitachi 2460N SEM. But we typically run several thousand counts per second for most of our samples. If we turn our beam off, we can still get our strobe peak. Therefore, I wonder if the issue is one of count rate. What is your typical rate. I am not familiar with a DSM960a so I don't know what your normal conditions are. I hope this helps some.
At 08:47 AM 5/6/97 -0700, you wrote: } Hello, } I have a Zeiss DSM 960a with an Oxford Link EDS system. The EDS spectra } has a large noise peak, typically 10 times the dominant real peak. I } have been talking with Oxford about this a great deal and they are } unsure of the cause. I now ask you for input. } } I checked the electrical isolation between the dewar and the column and } found there is none (1 Ohm) if the cables are connected and infinite if } they are disconnected. To Oxford's suprise they found the same on their } system. Is this normal? This means that noise in the SEM is in the } EDS!? How should I be grounding this system. We do have a 1 cm } diameter copper wire that is a dedicated ground running up to these to } systems. Any general tips on electrical issues for EDS would be } appreciated. } } thanks } } Brad Storey } Materials Scientist } Argonne National Lab - West } P.O. Box 2528 } Idaho Falls, ID 83403 } Ph. 208-533-7685 } Fax 208-533-7683 } e-mail brad.storey-at-anl.gov ---------------------------------------------------- Warren E. Straszheim 270 Metals Development, Ames Lab/ISU, Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-3091
} I have a Reichert Ultracut E that needs service. Does anyone know } who does repairs on these things? I'm on the west coast, is there someone } local?
Paul Swerdlick, of Leica Inc.'s service group, is based in California. He recently worked on our two Ultracut E's. Contact me privately for his phone number, if you need it.
} Keith Porter died last Friday. For those of you who aren't familiar with } this name, Porter truly deserved the title 'pioneer'. While he was at the } Rockefeller University, he helped develop the use of electron microscopy } for biological materials. Thus, together with other giants like George } Palade, he discovered cellular organelles that we now take for granted, } like the endoplasmic reticulum. He also helped invent the microtome } with which ultrathin sections could be cut. For years it was called the } Porter-Blum ultramicrotome. After the introduction of glutaraldehyde as } a fixative, he helped discover microtubules and thus played a key role in } igniting interest in the cytoskeleton. While his work primarily } centered on animal cells, Porter also investigated a variety of other } organisms, including plants. Together with Myron Ledbetter, he discovered } cortical microtubules in plant cells and therefore helped establish the } microtubule-cellulose microfibril paradigm. } Porter literally helped start the modern era of cell biology. He } moved from Rockefeller to Harvard and attracted a generation of new } scientists to the emerging field including Peter Hepler, Ben Bouck, Dick } McIntosh, and Lew Tilney, all of whom do or have worked on plants and } algae. Our own David Porter was an alumnus of the Keith Porter lab. Later, } Keith switched to the University of Colorado to found the Department of } Molecular, Cellular and Developmental Biology, one of the first if not the } first department of its kind. He thus started another trend: we now have } many departments of molecular and/or cellular biology all around the } world. } Porter was also president (and I believe a founder) of the } American Society for Cell Biology, one of the most active scientific } societies in the world, and he helped start the Journal of Cell Biology. } He also received the National Medal of Science. Sadly, when the Nobel } Prize in Medicine and Physiology was handed out years ago to George } Palade, Albert Claude and Chistian DeDuve for pioneering studies on cell } ultrastructure, Porter was snubbed. It was a great injustice. } Keith Porter was also known for his wit and sarcasm. Having } witnessed his sparkle from meeting halls and to elevators, I can tell you } that he was one of a kind. } In his last years, Keith was an emeritus professor at the } University of Pennsylvania. I am told that he was close to Kenneth } Thimann, a legend in plant physiology (identified indole acetic acid } as auxin) who also spent his last days at Penn and who, coincidentally, } also recently passed away. } Keith Porter's obituary appears in today's New York Times } (www.nytimes.com).
This message was sent to everyone in my Department today. Thought I should share it with y'all.
Best regards, Beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
One of my students, a talented young boy finishing his third year of studying Materials Engineering is looking for a job for the summer. He is ready to do anything for two months at a university or research institute abroad, speaks English quite well, and is experienced in SEM as well as other analytical techniques. He will be satisfied if he can pay his accommodation and living expenses while working since he just wants to gain experience and face the challenge. Please answer me directly and not through the List.
Thanks,
Kris
************************************************************************* Dr. Kristof KOVACS Associate Professor University of Veszprem Department of Silicate and Materials Engineering, Central Laboratory P.O.Box 158, Veszprem, HUNGARY H-8201 Phone: +36-(88)-421-684 *************************************************************************
Has anyone used those knife cleaners that swirl water back and forth? Does anyone have a better idea? Even trying to keep the knife clean I seem to get pieces of resin on the back side that is dificult to get off with a jet streem of water from a bottle or syringe. Thanks in advance.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com -----------------------------------------------------------------------.
How does one go about getting added (and deleted) from your mailing list?
MG
In case you didn't receive a copy of the message, the answer is found above.
Brad Storey wrote: } } Hello, } I have a Zeiss DSM 960a with an Oxford Link EDS system. The EDS spectra } has a large noise peak, typically 10 times the dominant real peak. I
} have been talking with Oxford about this a great deal and they are } unsure of the cause. I now ask you for input. } } I checked the electrical isolation between the dewar and the column and } found there is none (1 Ohm) if the cables are connected and infinite if } they are disconnected. To Oxford's suprise they found the same on their } system. Is this normal? This means that noise in the SEM is in the } EDS!? How should I be grounding this system. We do have a 1 cm } diameter copper wire that is a dedicated ground running up to these to } systems. Any general tips on electrical issues for EDS would be } appreciated. } } thanks } } Brad Storey } Materials Scientist } Argonne National Lab - West } P.O. Box 2528 } Idaho Falls, ID 83403 } Ph. 208-533-7685 } Fax 208-533-7683 } e-mail brad.storey-at-anl.gov }
Dear Brad, I think the noise can occur for four reasons:
1. Microphone effect from small crystals of ice, which accumulated at the bottom of dewar. They can be seen at the bottom in a moment when the nitrogen in dewar is almost over. I got rid of them with the help of some litres of boiling water, which filled in instead of liquid nitrogen. For realization of this procedure it is necessary to take off the detector from microscope, to pour out nitrogen, to fill boiled water and shake up approx. 10 seconds. Then it is necessary to dry up the dewar with the help of compressed air or other similar gas. All operations should be done quickly, that the sensor plate and FET transistor inside the detector had no time to heat up.
2. Microscope and EDS are connected to different phases of 3-phases power mains. Check they are connected to the same phase. If microscope uses all three phases, EDS and low voltage electronics of the microscope should be connected to the same phase.
3. Grounding. The detector and column should be electrically isolated from each other in order to be grounded only in one point. Your situation is OK. In general the good grounding is complex technical problem. Sometimes it is better (in sense of noise) to work with neutral connection instead of grounding (but all metal subjects in a room should be connected equally), sometimes without it at whole (but it is dangerous for health!). It is very difficulty to advise something about grounding from far away.
4. Dust on high-voltage circuits. Sometimes a dust accumulates on high-voltage circuits, then microdischarges of high voltage appear. These microdischarges cause occurrence of noise on a spectrum. Therefore these HV circuits need sometimes to be cleaned. They are located inside the rack with electronics and in preamplifier, which hangs on dewar. Caution! It is necessary to wait about half-hour after turning off the instruments until the charge flows down.
Attention! All above works should be carried out by the qualified personnel.
} Hello, } I have a Zeiss DSM 960a with an Oxford Link EDS system. The EDS spectra } has a large noise peak, typically 10 times the dominant real peak. I } have been talking with Oxford about this a great deal and they are } unsure of the cause. I now ask you for input.
Ground loops can be a source of major noise peaks such as this, but I have seen them caused both by faults in the preamp, or incorrectly adjusted preamps (some EDS preamps have trim pots for adjustment - don't touch unless you have detailed instructions for set up)
} I checked the electrical isolation between the dewar and the column and } found there is none (1 Ohm) if the cables are connected and infinite if } they are disconnected. To Oxford's suprise they found the same on their } system. Is this normal?
I would be worried about this, because it is exactly as it should be - find somebody else at Oxford to talk to:)
} This means that noise in the SEM is in the } EDS!? How should I be grounding this system. We do have a 1 cm } diameter copper wire that is a dedicated ground running up to these to } systems.
The EDS detector should be grounded through its control electronics, the power supply of which then goes back to the same source as the SEM this is where you dedicated ground should connect in. Make sure the grounding scheme is 'singly connected', that is any point in the syste can only get to ground by one route - you may have to spend a lot of time disconnecting and one-by-one reconnecting various accessories.
} Any general tips on electrical issues for EDS would be appreciated. } } thanks } } Brad Storey } Materials Scientist } Argonne National Lab - West
I'd first suggest taking the detector off the SEM and powering it all up on the bench. Its nice to have a radioactive source to allow you to test the EDS on the bench. If the noise peak disappears, it's got to be something to do with the SEM, although if as you say the EDS detector is correctly isolated from the SEM, I'd expect the peak to stay there.
You don't actually specify where you see the noise peak - I'm assuming it is right at the low energy cut off of the spectrum. I'm not familiar with the latest systems but certainly with earlier units, there is always a noise peak at the low energy end - it is usually not apparent because it is electronically cut off at the preamp - one of the trim pots I mentioned earlier. I've also seen systems where the output from the noise peak seems to swamp the preamp in some way, so that as you adjust the position of the low end cut off, and remove the noise peak, the count rate in the real peaks goes up.
Brad Storey wrote: } } Hello, } I have a Zeiss DSM 960a with an Oxford Link EDS system. The EDS spectra } has a large noise peak, typically 10 times the dominant real peak. I
} have been talking with Oxford about this a great deal and they are } unsure of the cause. I now ask you for input. } } I checked the electrical isolation between the dewar and the column and } found there is none (1 Ohm) if the cables are connected and infinite if } they are disconnected. To Oxford's suprise they found the same on their } system. Is this normal? This means that noise in the SEM is in the } EDS!? How should I be grounding this system. We do have a 1 cm } diameter copper wire that is a dedicated ground running up to these to } systems. Any general tips on electrical issues for EDS would be } appreciated. } } thanks } } Brad Storey } Materials Scientist } Argonne National Lab - West } P.O. Box 2528 } Idaho Falls, ID 83403 } Ph. 208-533-7685 } Fax 208-533-7683 } e-mail brad.storey-at-anl.gov }
Dear Brad, I think the noise can occur for four reasons:
1. Microphone effect from small crystals of ice, which accumulated at the bottom of dewar. They can be seen at the bottom in a moment when the nitrogen in dewar is almost over. I got rid of them with the help of some litres of boiling water, which filled in instead of liquid nitrogen. For realization of this procedure it is necessary to take off the detector from microscope, to pour out nitrogen, to fill boiled water and shake up approx. 10 seconds. Then it is necessary to dry up the dewar with the help of compressed air or other similar gas. All operations should be done quickly, that the sensor plate and FET transistor inside the detector had no time to heat up.
2. Microscope and EDS are connected to different phases of 3-phases power mains. Check they are connected to the same phase. If microscope uses all three phases, EDS and low voltage electronics of the microscope should be connected to the same phase.
3. Grounding. The detector and column should be electrically isolated from each other in order to be grounded only in one point. Your situation is OK. In general the good grounding is complex technical problem. Sometimes it is better (in sense of noise) to work with neutral connection instead of grounding (but all metal subjects in a room should be connected equally), sometimes without it at whole (but it is dangerous for health!). It is very difficulty to advise something about grounding from far away.
4. Dust on high-voltage circuits. Sometimes a dust accumulates on high-voltage circuits, then microdischarges of high voltage appear. These microdischarges cause occurrence of noise on a spectrum. Therefore these HV circuits need sometimes to be cleaned. They are located inside the rack with electronics and in preamplifier, which hangs on dewar. Caution! It is necessary to wait about half-hour after turning off the instruments until the charge flows down.
Attention! All above works should be carried out by the qualified personnel.
} } I have a Zeiss DSM 960a with an Oxford Link EDS system. The EDS spectra } } has a large noise peak, typically 10 times the dominant real peak. I } } have been talking with Oxford about this a great deal and they are } } unsure of the cause. I now ask you for input. } } } } I checked the electrical isolation between the dewar and the column and } } found there is none (1 Ohm) if the cables are connected and infinite if } } they are disconnected. To Oxford's suprise they found the same on their } } system. Is this normal? This means that noise in the SEM is in the } } EDS!? How should I be grounding this system. We do have a 1 cm } } diameter copper wire that is a dedicated ground running up to these to } } systems. Any general tips on electrical issues for EDS would be } } appreciated. } }
The dewar must be electrically isolated from the column or you will get a ground loop between the EDS electronics and the microscope grounds. The ground path must be via the pre-amp cabling, so this is correct. I'm amazed that the Oxford people that you are talking to did not know this, it's one of the things that is checked when you install a detector. For general grounding rules, think meca, all grounds must have one path to the main ground. If you have two paths, then that's a ground loop.
Sounds like your pulse processor's discriminators are not adjusted correctly. You need to monitor the input or fast channel cps to adjust the discriminators. The adjustment is easy, much better to tune than Noran or Kevex pulse processors. Check your manual for how to adjust the pulse processor (if you don't have one, get Oxford to FedEx you one, you should have it). You will either have a 2010, 2020, 2040, XP2 (computer adjusted), or XP3 (same as XP2 but standalone and manual adj). The new digital (DXP50) is most likely computer adjusted too).
The zero strobe goes off at about 500-600 cps. The zero strobe rate will decrease as the count rate is increased. It should be at zero eV in energy. With the beam off, it's all you should see (or rather 1/2 of the peak since the peak should be at zero eV).
Scott
----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 (tel) 4pi Analysis, Inc. Fax: 919 489-1487 (fax) 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com Durham, North Carolina 27707-2534 web: http://www.4pi.com
The Electron Microscopy Laboratory of New Mexico State University announces an opening for an Electron Microscopy Specialist. Qualifications: a B.S./M.S. degree with at least 4 years of experience in electron microscopy. The ideal candidate should have advanced experience in SEM/EDX. Proven experience in digital image analysis is a plus. The candidate should also be comfortable with the following techniques: metal evaporation, sputter coating, critical point drying, support film production, low temperature embedding, and film processing and printing. The successful candidate must be able to work well with people both as a collaborator and as a teacher. Duties include: record keeping, fixation, embedding, ultra-thin sectioning, staining, operation and routine maintenance of transmission and scanning electron microscopes and other laboratory equipment. Salary: $28,000 to $30,972. Benefits include: group medical and hospital insurance, group life insurance, state educational retirement, workmen's comp, sick leave, and unemployment compensation. Deadline for Applications: screening of applications will begin May 15 and will continue until a candidate is chosen. Applications should include a resume, letter of interest and 3 letters of recommedation. Send applications to: Arts and Science Research Center Box 30001/Dept. RC Las Cruces, NM 88003-8001 Tel. (505) 646-2611 Fax (505) 646-6095
I omitted the name of the person who wrote the article on Keith Porter. Dr. Barry Palevitz wrote the email that I forwarded to the list yesterday. I apologize for that omission.
Beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
Does anyone know of a commercial source of antibodies which could be used to identify golgi activity. My specimens ( P. carinii ) have been fixed with 4% paraformaldehyde - 0.1 % glutaraldehyde and are embedded in LR white resin. You can send replies directly to me at: mgoheen-at-indyvax.iupui.edu.
Thanks
Mike Goheen Dept of Pathology Indiana University School of Medicine
} ... } } I have a Zeiss DSM 960a with an Oxford Link EDS system. The EDS } spectra } } has a large noise peak, typically 10 times the dominant real peak. } ...
Noise can arise from nitrogen boiling in the dewar ... which can be an indication the vacuum in the dewar has degraded. Ice boining around in the dewar can also cause noise ...
cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
To everyone on this listserv, I have reposted the position for EM Specialist just to get those who might have missed it while on vacation or were glued to their tem binoculars. I am sorry if I have confused anyone.
I recently came across a procedure that Hildy Crowley gave me at a M.S.A. meeting some time ago in which the lead stain is kept in a 60 ml syringe that is fitted with 2 Acrodisc filters...and kept refrigerated. What I don't know is how often do I need to change out those filters. It would seem to me that a precipitate would form that would eventually cause artifact in the stain, showing up like "black boulders" under the TEM. Has anybody used this system and how long does the stain stay "good"?
Thanks in advance,
Sharron G. Chism HT (ASCP) Electron Microscopy Lab Harris Methodist Hospital Fort Worth, Texas
A colleague of mine mentioned seeing an article recently (1-4 weeks ago) in The Chronicle of Higher Education regarding economy measures and electron microscopy units in universities/colleges. Does anyone have the reference so I can check it out in the library? I can imagine what it says .......
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 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 Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sharron, I use the same system. I have a syringe needle on the filter which I change daily, my naive thinking being that any precipitates will go with the needle and when the syringe is empty I start afresh with a 20ml syringe and 0.2=B5m nylon Acrodisc filter. I use the same system for uranyl acetate (saturated in 50% methanol), this time with a 0.45=B5m Acro LC3S filter. I've been using this system for years and it's never let me down giving clean crisp staining. Ian.
I am collecting information on the currently valid standards applied in metallographic examinations in different countries and corresponding to the following standards of ASTM, ISO nad JIS:
In april 20-24 1998 in Amsterdam, the Netherlnads is made "7th European Congress For Strereology" title "trends and challenges in stereology"
adres local organizing committee:
Congress Secretariat VU Conference Service De Boelelaan 1105 1081 HV Amsterdam The Netherlands tel: +31 (0)20 4445790 fax: +31 (0)20 4445825 e-mail:vu_conference-at-dienst.vu.nl
Krzysztof Jan Huebner
{hubner-at-IOd.krakow.pl} :-)
FOUNDRY RESEARCH INSTITUTE Research Materials Department Structural and Physical Research Laboratory
We have embedded bacteria in LR White successfully but now are trying to embed a fungus, Ustilago hordei, so that we can do immunogold work with it. We had embedded it for years in various epoxies and had few problems, but with the LR White we have large holes not only in the cells but outside, not random but so that it appears that the cells are loose in the acrylic instead of embedded in it. Does anyone have experience preparing similar materials for immuno work? We don't have the equipment for frozen sections. Joyce Craig Chicago State University jcraig-at-csu.edu
I am working on a potato storage project and trying to conquer embedding of amyloplast starch in tuber samples. I've been using Spurr's resin, after 2% glutaraldehyde fixation in sodium cacodylate buffer and osmium tetroxide post fixation. I use propylene oxide after ethanol dehydration series to ensure dry samples, but still most of the starch granules and plastids fall out during sectioning and post staining.
Has anybody else worked with starch or potato tuber with success? I'm looking for any protocol hints that will better preserve starch and amyloplasts. Has anybody tried microwave fixation procedures on starch samples?
Thanks for any help you can give. I'll send 5# of the fantastic Minnesota russets to anyone who provides information that leads to success (but you'll have to wait until October to get them!).
Darryl Krueger Mn. Ag. Experiment Station EM Lab Univeristy of Minnesota St. Paul, MN 55108 (612)625-8249 darrylk-at-puccini.crl.umn.edu
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Subject: Time:1:14 PM OFFICE MEMO POSTDOCTORAL POSITION - ELECTRON... Date:5/8/97
POSTDOCTORAL POSITION - ELECTRON MICROSCOPY OF ELECTRONIC OXIDES
A post-doctoral position is available for electron microscopy studies of electronic oxide materials. The materials of interest include high-temperature superconductors, diffusion membranes, and colossal magneto-resistive compounds.
A Ph.D. in Materials Science/Physics or related area is required. Candidates with a strong background in interfaces, crystallography, and the effects of crystal defects on properties, especially in oxides, are preferred as are those with experience in one or more of the materials systems described above. Extensive hands-on experience in both analytical electron microscopy and HREM is required as is a demonstrated proficiency in TEM specimen preparation of bulk and thin film samples.
Additional skills in x-ray diffraction, in-situ experimentation in either EM or XRD, or measurement of electrical and/or ionic transport properties are also highly desirable.
Interested candidates should send a resume, publication list, and the names of three references to:
Dean Miller Materials Science Division Argonne National Laboratory Argonne, IL 60439
Yes, you read it right! FREE MONEY!!!!! Up to $600 per day!! How can that be possible, you ask? A new money making concept generates income for YOU, automatically.
Do you ever wonder how the wealthy, successful people get all their money? Now you can find out how they do it!!!! The secret is AUTOMATIC INCOME.
You say it takes money to make money. NOT ANY MORE!!!! No start- up costs, no personal selling, no obligation, no one will call you, EVER!!! Just FREE MONEY each and every month!!!
To receive a FREE copy of the report "FREE MONEY", do not hit Reply. Instead, simply e-mail your name and postal address to "Starind-at-neo-quest.com". Insert "FREE MONEY" in the subject heading.
(PS) Free money goes fast, so don't delay. Position and timing are everything!!!
Yes, you read it right! FREE MONEY!!!!! Up to $600 per day!! How can that be possible, you ask? A new money making concept generates income for YOU, automatically.
Do you ever wonder how the wealthy, successful people get all their money? Now you can find out how they do it!!!! The secret is AUTOMATIC INCOME.
You say it takes money to make money. NOT ANY MORE!!!! No start- up costs, no personal selling, no obligation, no one will call you, EVER!!! Just FREE MONEY each and every month!!!
To receive a FREE copy of the report "FREE MONEY", do not hit Reply. Simply e-mail your name and postal address to "Starind-at-neo-quest.com". Insert "FREE MONEY" in the subject heading.
(PS) Free money goes fast, so don't delay. Position and timing are everything!!!
The Goodyear Tire & Rubber Company, a world leader in tire development and manufacturing, has an excellent opportunity for a professional skilled in Microscopy in its Analytical Services Department, Corporate Research Division, located in Akron, Ohio. This position involves the analysis and characterization of tire components, polymers and other materials used in the rubber industry.
Qualified candidates must have a Ph.D. or MS in Analytical Chemistry with experience in stereo light microscopy and photomicrography. Strong skills in the implementation of computers in an analytical laboratory and experience in custom application development are also required. A working knowledge of statistics, chromatography and infrared spectroscopy is highly preferred.
This position, with a Fortune 100 industry leader, offers excellent benefits, relocation assistance and competitive salary commensurate with education and experience. If you have the qualification and desire to meet the rewarding challenges presented by Goodyear, direct your resume to:
J. A. Kutsko THE GOODYEAR TIRE & RUBBER COMPANY 1144 E. Market Street Akron, Ohio 44316
An Equal Opportunity Employer, M/F/D/V Applicants must be lawfully authorized to work in the U.S.
------------------------------------------------------------------------- John A. Wise jawise-at-goodyear.com The Goodyear Tire & Rubber Co. Research Analytical Services - D415B Voice 216.796.8716 142 Goodyear Blvd Fax 216.796.3304 Akron, OH 44305 U.S.A. -------------------------------------------------------------------------
We've down potato tuber by extending 4 to 5X the amount of time spent in each of the dehydration/ infiltration steps. Also, you might give a try using LR White. In the end we find there're still some problems as you mentioned but greatly reduced.
} Darryl wrote: } I am working on a potato storage project and trying to } conquer embedding of amyloplast starch in tuber samples. } I've been using Spurr's resin, after 2% glutaraldehyde } fixation in sodium cacodylate buffer and osmium tetroxide } post fixation. I use propylene oxide after ethanol } dehydration series to ensure dry samples, but still most } of the starch granules and plastids fall out during } sectioning and post staining. } } Has anybody else worked with starch or potato tuber with } success? I'm looking for any protocol hints that will } better preserve starch and amyloplasts. Has anybody tried } microwave fixation procedures on starch samples?
} Darryl, You might try a graded series of P.O. and Spurr and extend your infiltration times. I have used this on bone and it works fine. Or try Epon 812. I know it is viscous but it worked on yeast where Spurr did not. Best of luck!
Gregory Rudomen University Microscopy Imaging Center S.U.N.Y. Stony Brook Greg-at-UMIC.SUNYSB.EDU 516-444-3126
Those of you in the colleges and universities throughout the world are likely struggling with the task of grading final exams about this time of the year, while most of the rest of us will be rejoicing about the fact that we no longer have to take them, and so I thought all of you might enjoy this answer to a history exam question that was printed in the Winter 97 issue of The Hexagon, published by the Chemistry Honors Society, Alpha Chi Sigma. They didn't give the wording of the question exactly, but here's the answer:
"Renaissance was an age in which more individuals felt the value of their human being. Martin Luther was nailed to the church door at Wittenbert for selling papal indulgences. He died a horrible death, being excommunicated by a bull. It was the painter Donatello's interest in the female nude that made him the father of the Renaissance. It was an age of great inventions and discoveries. Gutenberg invented the Bible. Sir Walter Raleigh is a historical figure because he invented cigarettes. Another important invention was the circulation of blood. Sir Francis Drake circumcised the world with a 100-foot clipper."
How'd you like to have to assign a grade to that?
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
I have a query for this renowned group of microscopists. In attempting to account for time spent in the lab in response to" why isn't he sitting in front of the microscope more often" from a remotely situated administrator/bean counter, I thought I'd calculate a general (or range) ratio for the number of hours spent on the microscope (TEM) to the number of hours spent processing, sectioning (and semi-thinning? to find the block with the appropriate region), staining (conventional), development of negs/ printing (or digitally printing) cataloguing/marking the negs and prints, sending prints off and possibly writing reports, descriptions then phone calls, etc.on biological specimens (plant and animal tissue). I realize that plant material usually entails longer processing times and beam time (as it usually is 95% empty), but an average would be ok for plant and animal. If you do the final printing and plate prep for publications, add that in also. If you have just a couple of minutes to quickly compute the ratio of beam time to the pre/post time spent to completion I would appreciate your estimate . TIA, Hank Adams EML NMSU
Dear all, Thanks to all who replied to my recent queries to the list regarding maleate buffered-uranyl actetae enbloc stain and negative scanners.
My colleagues and I appreciate everyones suggestions and comments,
Yours,
----------------------------------------------------------------------- Richard Lander Senior Technician South Campus Electron Microscope Unit c/- Pathology Department Otago Medical School P.O. Box 913 Dunedin New Zealand. Tel. National 03 479 7301 Fax. National 03 479 7254
"Southernmost EM Unit in the World!" ------------------------------------------------------------------------
It seems to be generally known that perfluorinated-polyether pump oils and vacuum greases (e.g., Krytox, Fomblin) can cause micro-discharges on high-voltage insulators and thus lead to high-voltage instabilities. (This despite the fact that their beam-degradation mechanisms produce volatile fragments which should be pumped out of the system.)
Though I've gotten the above information from several authoritative sources (such as Wil Bigelow's book) I've never heard an explanation. Specifically: (1) What is the reason why these compounds create more of a high-voltage problem than normal hydrocarbon compounds?; and (2) What is the contamination mechanism? (Direct transfer of the compounds themselves? Deposit of molecular fragments? )
Microscopists: Perhaps you would like to flame that "free money" moron. I have send him a choice message. Apparently he is using our server "ultra.net" and I have asked the service provider to use a little influence too. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 350+ Links, MSDS ************************ http://www.proscitech.com.au
---------- } From: friend-at-noreply.com.ultra.net.au } To: friend-at-noreply.com.ultra.net.au } Subject: FREE MONEY! } Date: Friday, 9 May 1997 3:00 } } Dear Friend, } } Yes, you read it right! FREE MONEY!!!!! Up to $600 per day!! How can that be } possible, you ask? A new money making concept generates income for YOU, } automatically. snip . . . . .
You could try fixing with 2 - 3% potassium permanganate (aqueous) which gives excellent fixation and staining of amyloplasts - at least it did for root tips and endosperm of wheat. However my notes say not to embedd in Spurr's. We used araldite but I should imagine Epon would be OK.
my understanding of LR white is that, like all acrylics (I'm sure someone will correct me if I'm wrong), it does not cross polymerise well. This is one of it's advantages for immuno work because it should cause less damage to antigens than the epoxide resins. However it also tends to mean that it is less effective in embedding more difficult specimens and is less stable in the beam. I would suggest that, if you do not already do so, you should use coated grids to support the less stable sections and it may solve some of your problems.
I apologise if you know all this but it is a problem I've had for years - basically I use LR White for routine embedding of bacteria and cell pellets, and immuno work but I still use Spurr's for virtually everything else because it is reliable and versatile, despite the hazards.
Malcolm Haswell University of Sunderland UK ----------
Darryl Try the following it works in our lab.
Fix in 3% Glutaraldehyde in 0.05M sodium cacodylate buffer. (8 hours or overnight in fridge.)
2% osmium in 0.05M sodium cacodylate buffer - 2 hours maximum 3 x 30 minute wash in 0.05M sodium cacodylate buffer Block stain 2% aqueous uranyl acetate- 45 minutes 2 x 10 minutes wash in double distilled water Dehydration in ethanol series 10 minutes each. 2 x 30 minutes in propylene oxide
Infiltration - use epon/araldite resins To make up. 1 PART EPON 812 1 PART ARALDITE CY212 3 PARTS DDSA ( DODECENLY SUCCINIC ANHYDRIDE) Stir well with a glass stirrer.
25% resin* - 75% propylene oxide - 2 hours 50% resin* - 50% propylene oxide - 2hours 75% resin* - 25% propylene oxide -overnight with caps off in a fume cupboard 100% resins* -24 hours (2 changes in between)
POLYMERIZATION 100% RESIN IN ALUMINIUM DISHES OR EMBEDDING MOULDS IN A OVEN AT 70 DEGREES CENTIGRADE FOR 48 HOURS.
Very important : *Remember to add 1 drop of (DMP-30) 2,4,6 tri (dimethyl aminomethyl) phenol, per 1 ml epon each time. ie. every step for infiltration (25% -100%) If you miss out DMP-30 you might have to discard your specimen and start all over again.
All our fixation and infiltration steps are done in a glass pill vials in a fume cupboard.
I have also tried spurrs resins and found epon to be the best.
You can extend your infiltration time at 100% resins. Best of luck
Vijay H. Bandu Centre for Electron Microscopy Private Bag X01 Scottsville 3209 Kwa Zulu Natal South Arfica E-Mail bandu-at-emu.ac.za
Hi Darryl; My experience with potatoes is strictly at the light microscopy level, embedding in glycol methacrylate. While I realise that GMA isn't suitable for EM, there are, I believe, acrylics that are, such as LR White. My samples were fixed in 4% glutaraldehyde in phosphate buffer, dehydrated through a solvent series (not one I think you'd like for EM, but an ethanol series should do fine) and infiltrated in methacrylate for a week or so in the fridge. Starch granule preservation is quite nice, and I have even been able to observe the "growth rings" in various preparations. I do realize that phosphate buffer is not compatible with EM preps... but I doubt if the buffer would be your problem.
Good luck shea
Dr. S. Shea Miller Agriculture & Agri-Food Canada Eastern Cereal & Oilseed Research Centre Rm 2068, Bldg 20, CEF Ottawa, Ontario Canada K1A 0C6 Phone: (613)759-1760 Fax: (613)759-1701 e-mail: millers-at-em.agr.ca
I am Resia Pretorius from South Africa and am currently working on a PhD )the phylogeny of internal structures of the superfamily Scarabaeoidea ((insects))
I would like to use a SEM to photograph one of the internal chitinous structures (metendosternites) to which flight and leg muscles are attached, and do calculations of different lengths for morphometric analysis.
These muscles are attached firmly to the metendosternites and I would like to remove them, but so far without luck. I have tried Sodium perborate (different concentrations) they use this chemical to clean muscles from snake skulls, but if only loosens the muscles slightly, I still need to pull and tuck at the muscles to remove them.
I have also tried tripsin a 0.5% without it working. Are there any suggestions from anyone, I read in an 1890 article that one can use nitric acid for the macercation of muscles but that was obviously before the SEM was invented. Can one use an acid or would it harm the actual structure of the metendosternites.
I realy need help!! my e-mail address is:
resia-at-mcd4330.medunsa.ac.za
Thanks, Resia Pretorius Lecturer (Biology) Medunsa South Africa
I am doubtful that its a true infiltration problem. Poor fixation emulates infiltration problems. I assume that you are fixing in the cold and that will not do for most yeasts and plants with "massive" cell walls. Plants which store well generally are in that category.
Up to about 25 years ago most of such tissues were fixed in 1-4% KMnO4, without any buffers. Its a lousy fixatives and only preserves membranes, but try it for say 30 minutes at about 20 degrees. I expect that your sectioning and physical retention of starch and plastids will be much better.
To get better fixation I suggest to forget the GA, it does not penetrate those plant cell walls. Use the Osmium at at least 20 degrees and try 30, 60, 120 minutes.
Botanists working on storage tissues have for years struggled with fixation problems, mostly because they copied the biomedical "in ice" doctrine. Autolysis is much more rapid in animal cells than it is in plants. Keeping the temperature low dramatically slows enzyme reaction in animal cells and hence slows autolysis.
See it as a race: Low temperature slows fixation somewhat, but in animal cells much slower autolysis is the winner. In those "difficult" plant tissues cold fixation is disproportionately slower because of the massive cell walls. Additionally, plant enzyme's activities varies much less in the considered temperature range than would animal enzyme's activities. Accordingly, higher (20 - 35 degree) temperatures, rather than shorter fixation times are preferable for those "difficult" tissues.
I am sorry that I could not accept the offer of Minnesota russets, if offered. Australian Quarantine would have an instant baby, without a pregnancy what's more. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 350+ Links, MSDS ************************ http://www.proscitech.com.au ---------- } I am working on a potato storage project and trying to conquer embedding of } amyloplast starch in tuber samples. I've been using Spurr's resin, after 2% } glutaraldehyde fixation in sodium cacodylate buffer and osmium tetroxide post } fixation. I use propylene oxide after ethanol dehydration series to ensure dry } samples, but still most of the starch granules and plastids fall out during } sectioning and post staining. } } Has anybody else worked with starch or potato tuber with success? I'm looking } for any protocol hints that will better preserve starch and amyloplasts. Has } anybody tried microwave fixation procedures on starch samples? } } Thanks for any help you can give. I'll send 5# of the fantastic Minnesota
} russets to anyone who provides information that leads to success (but you'll } have to wait until October to get them!). } } Darryl Krueger } Mn. Ag. Experiment Station } EM Lab } Univeristy of Minnesota } St. Paul, MN 55108 } (612)625-8249 } darrylk-at-puccini.crl.umn.edu }
The wall of the fungus and may prevent good embedding. I have also had Aspergillus (esp. the spores) that would pull out if faced with a razor blade. We routinely sectioned these by doing all the block facing with a diamond knife (use an old one) and starting fully 50 microns above the sample. The sections were collected on formvar.
Need used SEM with W or LaB6 gun, prefer stepper motor stage.
................................................................. Dave Pierce pierce-at-magic.geol.ucsb.edu WB6LRN Geological Sciences (805) 893-2466 (voice/message) University of California (805) 893-2314 (fax) Santa Barbara, CA 93106
I often have trouble observing fine powder on my SEM. Powder that is less than 1 micron in size. I'll first either pack it lightly on carbon tape or sprinkle some over silver paint, then coat. When the electron beam hits it, I can see the powder being blown off and I get charging. What are some good techniques for observing light in weight and small powder on the SEM?
Mark E. Darus (216) 266-2895 General Electric Co. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I often have trouble observing fine powder on my SEM. Powder that is } less than 1 micron in size. I'll first either pack it lightly on carbon tape } or sprinkle some over silver paint, then coat. } When the electron beam hits it, I can see the powder being blown off and } I get charging. ...
The particle undersides are not apparently being coated ... but the problem is each particle sensing its neighbor and being repelled by it. I think if the particles were dispersed with increased distance, distributed individually on the carbon tape, you'd see "less effect" from charging ... Hope this helps ...
cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
} } I often have trouble observing fine powder on my SEM. Powder that is } less than 1 micron in size. I'll first either pack it lightly on carbon tape } or sprinkle some over silver paint, then coat. } When the electron beam hits it, I can see the powder being blown off and } I get charging. What are some good techniques for observing light in weight } and small powder on the SEM? } } Thanks } Mark Darus Dear Mark,
For our automated particle classification analysis by size, morphology and composition we disperse small amount of powder in appropriate liquid (water, oil, etc.) in ultrasonic bath and filter suspended particles onto 0.2 micron filter. Filter is transferred on a holder, coated, etc. Regards,
Igor C. Ivanov, SIMS,Auger,XPS,TEM,SEM Senior Scientist Analytical Services RJ LeeGroup, Inc. Contract Research 530 McCormick Str. (510)-567-0480 phone San Leandro, CA 94577 (510)-567-0488 FAX
I suggest you do the other way around: put a small amount of your powder on filter paper, then apply gently the SEM sample holder with carbon tape. Thus in theory only the particles in contact with carbon tape will adhere. In order to get rid of any other particle you may also want to blow dry nitrogen though I do not believe it is necessary, and if you don't, at least you'll be sure you won't get any contamination from N2. You must get a kind of monolayer of particles.
Then coat.
On Fri, 9 May 1997, Mark E. Darus wrote:
} I often have trouble observing fine powder on my SEM. Powder that is } less than 1 micron in size. I'll first either pack it lightly on carbon tape } or sprinkle some over silver paint, then coat. } When the electron beam hits it, I can see the powder being blown off and } I get charging. What are some good techniques for observing light in weight } and small powder on the SEM?
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Mark:
I'm not certain how applicable this is, but SPI Supplies offers a product called Tacky Dot Slides which, as I understand it, have some very small and precisely spaced "tacky dots" which will grab individual particles. You can probably get the information from their web site which I believe is at http://www.2spi.com. If I'm wrong about the web site, you can access it through my "related Links" section from my web site.
David Henriks TEL: 800-728-2233 (toll-free in USA) South Bay Technology, Inc. 714-492-2600 1120 Via Callejon FAX: 714-492-1499 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 "Mark E. Darus (216) 266-2895 General Electric Co." } I often have trouble observing fine powder on my SEM. Powder that is less than 1 micron in size. I'll first either pack it lightly on carbon tape or sprinkle some over silver paint, then coat. When the electron beam hits it, I can see the powder being blown off and I get charging. What are some good techniques for observing light in weight and small powder on the SEM?
Dear Folks, I have relished our interchanges regarding the situation with Pbcitrate and NaOH. Here is a short summary and some ideas. 1. Good citrate staining is the product of many contributions. If there are problems with it however, the simplest and most likely cause is the incorrect production of the stain. Therefore - 2. If you are happy with your stain, if it is reliable, and you never worry about it working, and you do not have to make it up often, leave it alone no matter how you make it. 3. If you are anxious about it, sometimes it stains, sometimes it is very good, other times marginal, and sometimes it may even precipitate, do the following simple things: Use commercially prepared 1 N NaOH instead of pellets, and shake and invert your volumetric for 25 minutes. Then store immediately in plastic syringes. 4. If you frequently come to a dead stop, frequently have sections ruined or unpublishable, it is time to get out the Reynolds paper and follow instructions carefully, but do not rinse your sections after staining in anything but plain dwater. 5. If the above does not improve the situation, there may be a major problem with your embedding process. If so, please call me at 303-871-3026, and I will be glad to see if I can help.
In summary it has always seemed to me that it is imperative that all basic operations of an EM lab such as processing, embedding, cutting, staining, scoping, photomic, must, must, must run flawlessly, simply, and reproducibly without worries, so that we can save our emotional and intellectual energies for our experimental conditions. Therefore I believe in making up quantities (500ml) of Pbcit with 1.0N NaOH (do not pH it) storing it in syringes in the refrigerator according to the method of Reynolds, and then using it for at least six months. I have been asked what the pH should be. I do not know. It is not important (and is probably counterproductive as it leads to using the pH meter which should not be done). I know absolutely that small differences in pH will change the staining properties of Pbcit. (Pellets are hard to weigh, they absorb water, etc, sometimes they work and sometimes they are off - it is just not worth fussing with them.) If someone has stubborn problems, please call me, and I will see if I can be of help. Bye now, Hildy
We do a lot of TEM of keratinocyte cell cultures using this procedure:
EMBEDDING PROCEDURE /cell culture plates
PBS rinsex3 (discard leftover tissue media down sink with bleach)
1/2 KARNOVSKYS 3hrs to overnight
0.1M NaCaco 15minx2
1%OsO4 1hr
dH2O 15minx2
1%UA 11/2 hrs
35%ETOH 15minx2
70%ETOH 15minx2
95%ETOH 15minx2
100%ETOH 15minx2
100% ETOH 30minx2
3:1 ETOH:EPON 6-8hrs
2:1 ETOH:EPON 12-16hrs (overnight w/ caps off)
1:1 ETOH:EPON 6-8hrs (w/ caps off)
100%EPON 6-8hrs
bake for 24-48 hours in 600 C oven
DO NOT USE PROPYLENE OXIDE!!! IT DISSOLVES THE PLASTIC TISSUE CULTURE PLATES
Bob Underwood Morphology Core Univ of Washington
On 8 May 1997, Manoj Misra wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Wonder if someone has a protocol for preparing keratinocyte } (skin cell) culture for TEM. } } Thanks, } } Manoj Misra }
I did a time study 10 years ago while working as an EM tech in a hospital pathology lab with a Siemens TEM. The pathologist attempted the scanning and photography at the microscope. Results: 16% tissue preparation 30% cutting and staining 25% photo processing 12% equipment maintenance 9% lab management 5% continuing education and professional activities 2% training, demonstration, public relations Final result: Closure of the lab and loss of my position, thanks to the bean counters.
Hi! Are there any specific stains that react only with nucleic acids that can be used for negative staining in transmission electron microscopy of plasmid DNA without additional shadowing with tungsten/platinum/palladium etc? Thanks, Yamini Dalal
On Fri, 9 May 1997, Mark E. Darus (216) 266-2895 General Electric Co. wrote:
} -----------------------------------------------------------------------. } I often have trouble observing fine powder on my SEM. Powder that is } less than 1 micron in size. I'll first either pack it lightly on carbon tape } or sprinkle some over silver paint, then coat. } When the electron beam hits it, I can see the powder being blown off and } I get charging. What are some good techniques for observing light in weight } and small powder on the SEM? }
I think it is a matter of conductivity on those unbond or loose particles. Normally I sprinkle the powder over sticky-tab on stub and use a Duster to blow those unbond particle away. After sputter coating you will get a beautiful image of particles without charging or being blown off. This is my two cents tip and wish you luck.
*********************************************** * Ming H. Chen, PhD * * Medicine/Dentistry Electron Microscopy Unit * * University Of Alberta. * * Edmonton, Alberta, Canada * * * * Visit My Page At: * * http://www.ualberta.ca/~mingchen * ***********************************************
Another variation: Drop the powder onto the carbon taped stub, then rap the side of the stub to make the powder "slide" around. Finally, dump/jar/blow the loose powder off the tape surface. This tends to also give a "mono-layer" of powder on the stub with fewer charging problems. Just for comparison I also usually press the powder down in one area with a small spatula blade before removing the excess (sort of a hedge against any weird settling phenomena occurring.) The pressed area virtually always charges much worse than the rest of the stub surface.
----------------------------------------------------- Bede Willenbring Research Chemist
H.B. Fuller Company 1200 Willow Lake Blvd Vadnais Heights MN 55110-5132
} } } Yves Maniette {yves-at-giga.sct.ub.es} 5/9/97 13:15 } } } I suggest you do the other way around: put a small amount of your powder on filter paper, then apply gently the SEM sample holder with carbon tape...
} ... I often have trouble observing fine powder on my SEM. Powder that is } less than 1 micron in size. I'll first either pack it lightly on carbon tape } or sprinkle some over silver paint, then coat....
} ... } Microscopists: Perhaps you would like to flame that "free money" moron. I } have send him a choice message. Apparently he is using our server } "ultra.net" and I have asked the service provider to use a little influence } too. } Jim Darley }
The domain given in the body of the "free money" message, neo-quest.com, is registered to the lowlife Sanford Wallace, of the detested cyberpromo.com, who makes a living spamming with false From: addresses. From a WHOIS search:
Neo-Quest NEO-QUEST-DOM 3900 Moorpark Ave.-Suite 27 San Jose, CA 95117 USA
Dear Mark, Whenever I observe powder in the SEM, I first determine the average size. For small powders like yours, the best method is to suspend it in a solvent such as alcohol, disperse by sonicating, then put a drop on a smooth surface, such as a glass coverslip or glassy carbon. Wait until it dries, then coat as usual. The powder is then thinly dispersed. Most of your problem is because the powder is piled up too high. I always say that if you can see it, its too thick. You wrote:
} I often have trouble observing fine powder on my SEM. Powder that is } less than 1 micron in size. I'll first either pack it lightly on carbon tape } or sprinkle some over silver paint, then coat. } When the electron beam hits it, I can see the powder being blown off and } I get charging. What are some good techniques for observing light in weight } and small powder on the SEM? } } Thanks } Mark Darus }
Regards, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
Dear Mark, when I worked on Auger microscope, I needed frequently to investigate of a non-conducting powder without any coating. Best it resulted if the powder is rolled up in In foil. Please work with In in gloves. Regards, Victor
Victor Sidorenko ANTRON Co. Ltd., scientific service Moscow, Russia antron-at-space.ru
Thank you to all of you who have answer to my question. It seems that no such a list exist on the web, anyway I got some useful information that I post below:
Response 1
If you are referring to metals, then there are several good books published by ASM (American Society for (Metals/Materials). There is the ASM Handbook, several guides to metallography as well as George Vander Voorts (?) guide to metallographic preparation and etchants.
Good Luck.
Alan Stone {as-at-mcs.com} ASTON
Response 2
There are a number of good reference books on sampe preparation for LM. I have listed a few of them below.
1) "Metallography & Microstructures" American Society for Materials, Volume 9
2) "Metallographic Polishing by Mechanical Methods" L.E. Samuals, American Society for Materials
3) "Metallographic Specimen Preparation" James L. McCall & William M. Mueller, Plenum Press ISBN 0-306-30791-x
4) "Metallography of Advanced Meterials" Henry J. Cialone et al, American Society for Materials.
You may also want to check with manufacturers of such equipment. We maintain a database of sample preparation methods and can often give references fr specific preparation needs. We also publish a bibliography of technical papers dealing with sample preparation which we are pleased to send you at no charge. You can then select papers of interest and we will send you reprints at no charge.
I hope this helps!
Best regards-
David Henriks (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.
Response 3
Look on the List web server . This server is located at: http://www.liszt.com/.
Henrik
-- Henrik Kaker SEM-EDS Laboratory, Metal Ravne d.o.o., Slovenia, mailto:Henrik.Kaker-at-guest.arnes.si
We've been doing this on Tingids for Dr Gerry Cassis here at the Australian Museum. He's in a team which includes experts on Crustacea, Arachnids, Trilobites, etc. who are looking at the evolutionary origins of the arthropod groups.
The final protocol worked out by Sue Lindsay was;
Digest the muscle tissues in either KOH or NaOH (3 pellets in 3 or 4 mls water in watchglass at 70 degrees C). Leaving it in this solution after the tissue has digested will also start to clear the exoskeleton.
Partially clear the exoskeleton for examination under transmitted light microscopy by either leaving it in the caustic solution for much longer or by BREIFLY boiling the solution. I'm told that Lactic Acid is also useful for clearing exoskeletons. Light microscopy was useful for detecting overlapping of sternites, relative thickness of sternites, difference between sternite and arthrodial membrane, etc.
Clean in ultrasonic bath for extended time (say 5 minutes) to get all the digested ooze out - obviously with some escape path such as the holes where the head &/or abdomen were.
Dry the specimen. We got our best results with Critical Point Drying, though this was because the tingids were a bit flimsy. Your scarabs might me more robust, in which case HMDS (HexaMethyl DiSilazine) might work well, or even just air drying from a volatile solvent (Acetone or Ethanol?).
Dissect the specimen. We use eye surgery scissors, very small blade length which worked on even the smallish specimens. Cut along the dorsal and ventral midlines for two mirror-image samples; in our case, one for SEM and the other for light microscopy.
Comments: We found the order to be important, since cutting the specimen first and then digesting the tissue lead to the skeleton just rolling up on drying. But this probably depends entirely on your particular specimen. If given the choice I'd rather cut first and digest second, making digestion quicker and allowing you to help it along by picking away at the muscle with forceps. The ultrasound cleaning would also be easier. As always in EM, you've just got to find what works with your particular beasts.
Geoff Avern Manager Microscopy Labs Australian Museum Sydney, Australia
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi,
I am Resia Pretorius from South Africa and am currently working on a PhD )the phylogeny of internal structures of the superfamily Scarabaeoidea ((insects))
I would like to use a SEM to photograph one of the internal chitinous structures (metendosternites) to which flight and leg muscles are attached, and do calculations of different lengths for morphometric analysis.
These muscles are attached firmly to the metendosternites and I would like to remove them, but so far without luck. I have tried Sodium perborate (different concentrations) they use this chemical to clean muscles from snake skulls, but if only loosens the muscles slightly, I still need to pull and tuck at the muscles to remove them.
I have also tried tripsin a 0.5% without it working. Are there any suggestions from anyone, I read in an 1890 article that one can use nitric acid for the macercation of muscles but that was obviously before the SEM was invented. Can one use an acid or would it harm the actual structure of the metendosternites.
I realy need help!! my e-mail address is:
resia-at-mcd4330.medunsa.ac.za
Thanks, Resia Pretorius Lecturer (Biology) Medunsa South Africa
Message-Id: {1.5.4.32.19970512034746.00693dbc-at-pop3.unsw.edu.au} X-Sender: s7001031-at-pop3.unsw.edu.au X-Mailer: Windows Eudora Light Version 1.5.4 (32) Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Tempfix is good for powders. Its an adhesive which melts around 80 deg. We melt some onto a stub, then warm it until the adhesive is just liquid, then press it into the powder. Let it cool, then blow with an air duster. If it blows off it wasn't stuck down. Most EM supplier stock it.
Hello dear subscribers I am a new comer to this listserver. I am interested to take training in Electron Microscopy. I have already taken preliminary training in EM in KKUH, and KSU. I am looking addresses of Institutions/Univresities/Organizations where I can undertake further training in Electron Microscopy . Pls send any information at my following email address: F40C028-at-KSU.EDU.SA I appreciate and be solicited for providing informations. Thanking you in anticipation. Abdulrahman N.S.Al-Ohali
I try to immunostain A431 cells with the preembedding method. Staining works well with 1nm gold and IGSS staining (IntenSE) on the LM level. On the EM level I can not detect any staining. Samples are fixed in glutaraldehyd and osmium after IGSS staining and conventionally dehydrated and embedded in Epon. It seems that during the fixation and embedding procedure the staining is washed out. Is there anyone who has made the same experience?
Dear Microscopists, I am working on the dermis of echinoderms on which I plan to perform a high-pressure freezing followed by a freeze-substitution in two steps: firstly in a solution containing glutaraldehyde and tannic acid and secondly in osmium tetroxide. I am wondering if I would better use tannic acid (i.e. a mixture of different gallotannins) or 3,4,5-Trihydroxy-benzoic-acid (C7H6O5), the active component of the so called tannic acid. Has anybody experience in the subject? I thank you for your help,
Uranyl acetate will stain DNA, but that is a positive stain and is not highly specific for nucleic acids. UA and several other heavy metal salts can be used as negative stains, but in that case you do not want the DNA to stain specifically; this is usually done when you want to visualize proteins associated with DNA (eg Griffith, J. Science 187: 1202 and 201: 525). DNA replicated in the presence of BrdU or biotin-labeled nucleotides can be specifically stained with anti-BrdU and gold-labeled secondary antibodies or avidin.
Also, DNA can be spread on a surface, picked up on a film, positively stained with UA, and embedded in a thin layer of the detergent photoflo.
-Dennis Goode
} Reply-to: lsdalal-at-scifac.indstate.edu Organization: ISU To: Microscopy-at-Sparc5.Microscopy.Com Subject: EM and DNA
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi! Are there any specific stains that react only with nucleic acids that can be used for negative staining in transmission electron microscopy of plasmid DNA without additional shadowing with tungsten/platinum/palladium etc? Thanks, Yamini Dalal Dr. M. Dennis Goode Phone (301) 405-6917 Department of Zoology Fax (301) 314-9358 University of Maryland e-mail goode-at-zool.umd.edu College Park MD 20742 ************************************************************* "If the Lord Almighty had consulted me before embarking upon the creation, I should have recommended something simpler." -Alphonso X of Castile, 15th Century
Thank you to all who responded to my query regarding slides imprinted with grid squares. Several people requested information be forwarded on, so the following is a quick summary of responses.
Field Finder Slides - a standard slide imprinted with grid squares intended to be used along with the (separate) specimen slide McCrone 'England Field Finder' 800-622-8122 Fisher 'Field Finder' 800-766-7000
Several coverslips were recommended, made by: Eppendorf 'cellocate' Bellco 800-257-7043 (USA) 800-445-7051 (Canada)
Thank you to IMR and McCann Imaging for their thoughtful and helpful advice as well as Charles Garber (SPI) and Stacie Kirsch (EMS) who pointed me to websites at: cccbi.chester.pa.us/spi/new/ptfesld.html (SPI) emsdiasum.com (EMS)
Thanks to all of you we now have the slides we needed. My regards to all of you who responded.
************************* * Marilyn Wadsworth * * mwadswor-at-zoo.uvm.edu * *************************
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
PHILADELPHIA SOCIETY FOR MICROSCOPY MAY 1997 MEETING NOTICE Wednesday, May 28, 1997
MICROSCOPY IN THE MUSEUM at the Philadelphia Museum of Art
The preservation of the collections for future generations is the first obligation of the Philadelphia Museum of Art as a public institution. The Museum's Conservation Department is entrusted with this responsibility. The Museum's staff of 17 conservators employ a wide range of analytical techniques for assessing the condition of objects in the collection as well as monitoring the success of treatments that are carried out. Many of these techniques are microscopy-based.
At this meeting, members of the conservation staff will give an overview of microscopy and other analytical techniques employed at the Philadelphia Museum of Art, which include PLM, visible light and UV microscopy, Micro-FTIR and SEM-EDS. Recent work ranging from the analysis of Czanne's paintings to William Penn atop City Hall will be discussed.
Following the presentations, there will be a "Behind the Scenes" tour of the Conservation and Analytical Labs. The Museum's galleries will be open for viewing as well, until 8:45 PM.
The Speakers: Marigene H. Butler Head of Conservation Andrew Lins Senior Conservator of Decorative Arts and Sculpture Beth Price Conservation Chemist Peter Eastman Conservation Information Specialist
*****PLEASE NOTE CHANGES FOR THIS SPECIAL EVENT*****
DATE: Wednesday, May 28, 1997
PLACE: Philadelphia Museum of Art Benjamin Franklin Parkway at Eakins Oval Philadelphia, PA
PARKING: Parking around the Museum will be difficult because of special Wednesday night events. Free parking may be available around the building before 5:30 PM. Additional parking can also be found along Pennsylvania Avenue (street level to the north), inside Eakins Oval (below the steps on the east side) or behind the Museum in the Azalea Garden (west side). There is parking and access for persons with disabilities along the south side of the Museum.
TIME: 5:15 - 6:30 PM Check-in. The check-in table will be located inside the West door (facing the Schuylkill River) during this time. You must check in to receive your Museum admission badge and pay for your meal. A Museum map of the galleries and the outdoor dinner tent will be provided.
5:30 - 7:00 PM Dinner will be served in the outdoor tent on the East Terrace. Members $12 Student members $6 Non-members $15
Menu: Penne pasta with roasted eggplant, red and yellow peppers and ricotta cheese Chicken provencal with olives, red and yellow peppers, zucchini and tomatoes Green beans, Roasted potatoes Sliced seasonal fruit Chocolate truffle cake Coffee, decaf or tea
6:55 PM Gather at check-in table before going to Conservation Lab
7:00 - 8:00 PM Presentations, in the Conservation Lab
8:00 - 8:30 PM "Behind the Scenes" tour through conservation laboratories
NEWS FLASH: Reservations can now be sent by e-mail by writing to PSM-RESERVATIONS-at-INAME.COM. Be sure to include your name and affiliation in the body of the message. PSM prefers that you use this convenient method to RSVP. If you don't have access to e-mail, phone reservations will be taken by Ms. Pat Overend at the University of Pennsylvania, 215/898-8337.
Deadline for reservations will be WEDNESDAY, May 21. At the Museum's request, we must insist that you adhere to this deadline. If you have any questions regarding the meeting please feel free to contact Rollin Lakis of the Executive Council T 215/898-8718. Cancellations must be received no later than 5:00 PM, May 23, 1997.
Bonus: The first 25 people to make reservations will receive complimentary tickets to the Rodin and Michelangelo exhibit, which will be open until 8:45 PM.
You are not alone. In fact this was talked about some time ago (last year?); check the archives for that discussion. I have had no luck with conventional embedding and IGSS. It seems that the osmium changes the Ag-Au which then washes out during UAc treatment. I've tried cold short osmium and alcoholic UAc, but it still happens. So I gold tone all my tissue before embedding. Adds contrast and the tissue looks "grainy" at high mag, but otherwise it's fine and the Ag-Au remains in place. The alternative (for me) is to use osmium only, no Uac, and increase the staining time with UAc on the grids. I find for my tissue that the contrast is still not good enough, but it may work on yours. If you want the gold toning method, Email me and I'll send it. Good luck.
Diana van Driel Dept Ophthalmology Sydney University C09 AUSTRALIA 2006
Dear All, I am currently using antibodies to localise fungi in semi-thin resin sections of Tomato. When using a gold conjugate system I have no problems, but I am also using WGA and another antibody both linked to Rhodamine, and there is a huge amount of autofluorescence from the plant and the fungus when observed under the green light. There is a similar problem with the filter blocks we have for FITC. I tried treating the sections with KOH solution and this worked really well for the WGA labelled sections reducing the problem, but obviously that is not an option with the antibody. Does anyone have any suggestions? Thanks,
Mark Munro. The Soil Biology Unit SAC e-mail m.munro-at-ab.sac.ac.uk.
Dear subscribers! I have several questions for the list. 1. Why everybody uses for section constrasting uranyl acetate which is less soluble than uranyl nitrate or uranyl sulfate? 2. Is fluorescence of green fluorescent protein (GFP) affected by low concentration of glutaraldehyde? 3. Is fluorescence of GFP affected after embedding into epoxy or acrylate resins? 4. Have epoxy and acrylate resins autofluoresence?
Sincerely yours, Alexander Mironov
Unit of Morphology Consorzio Mario Negri Sud S.Maria Imbaro (Chieti), 66030, Italy Fax: +39 872 578 240
Dear All, I am currently using antibodies to localise fungi in semi-thin resin sections of Tomato. When using a gold conjugate system I have no problems, but I am also using WGA and another antibody both linked to Rhodamine, and there is a huge amount of autofluorescence from the plant and the fungus when observed under the green light. There is a similar problem with the filter blocks we have for FITC. I tried treating the sections with KOH solution and this worked really well for the WGA labelled sections reducing the problem, but obviously that is not an option with the antibody. Does anyone have any suggestions? Thanks,
Mark Munro. The Soil Biology Unit SAC e-mail m.munro-at-ab.sac.ac.uk.
Dear All, I am currently using antibodies to localise fungi in semi-thin resin sections of Tomato. When using a gold conjugate system I have no problems, but I am also using WGA and another antibody both linked to Rhodamine, and there is a huge amount of autofluorescence from the plant and the fungus when observed under the green light. There is a similar problem with the filter blocks we have for FITC. I tried treating the sections with KOH solution and this worked really well for the WGA labelled sections reducing the problem, but obviously that is not an option with the antibody. Does anyone have any suggestions? Thanks,
Mark Munro. The Soil Biology Unit SAC e-mail m.munro-at-ab.sac.ac.uk.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America The important thing with Tannic acid is the original source. For E.M. the best is Turkish, available from Mallinckrodt and not the Chinese from Sigma. Ian.
Dear friends: Department of AME of Un. of AZ has received SEM S-650(Hitachi) as a gift. Howereve, there is't manual or documentation and we are not able run it. We would greatly appreciate your help in supplying us such manual or any information. Thanks. Yury Shipilov Research Assosiste ph.(520) 626-4470
Dear subscribers! I have several questions for the list. 1. Why everybody uses for section constrasting uranyl acetate which is less soluble than uranyl nitrate or uranyl sulfate? 2. Is fluorescence of green fluorescent protein (GFP) affected by low concentration of glutaraldehyde? 3. Is fluorescence of GFP affected after embedding into epoxy or acrylate resins? 4. Have epoxy and acrylate resins autofluoresence?
Sincerely yours, Alexander Mironov
Unit of Morphology Consorzio Mario Negri Sud S.Maria Imbaro (Chieti), 66030, Italy Fax: +39 872 578 240
I need to buy 2 new elemental standards blocks for use in our SEMs. Required dimensions are 1" diameter by { 1" thick. These must NOT be carbon coated. Any recomendations or products not to consider?
AccuMed International, Inc., has an immediate opening for a TECHNICAL = MANAGER to oversee the development, scientific and clinical testing, and = production of computerized, microscope-based IMAGING workstations for = clinical and research applications. The successful candidate will = shepherd new instruments from R&D to finished product. The position = requires a minimum of a Bachelor's degree and a strong = scientific/technical background, with at least 3-5 years experience in = developing and utilizing imaging workstations and applications for light = microscopy. Specific areas of responsibility will include:
- managing the development of new hardware and software products - establishing and maintaining timelines for delivery of prototype = instruments and finished products, including software-based applications - coordinating activities in various departments including R&D, = engineering, software development and manufacturing - assisting with applications to regulatory agencies
AccuMed International, headquartered in Chicago, is a global laboratory = diagnostic products company and an emerging leader in the field of = automated cytopathology. EOE.
Please respond via mail, FAX or email (NOT telephone) to:
Marc M. Friedman, Ph.D. Vice President, Scientifc and Technical Affairs AccuMed International, Inc. 900 N. Franklin, Suite 401 Chicago, IL 60610 Tel: (312) 642-9200 FAX: (312) 642-8684 email: marc.friedman-at-accumed.com
I am using immunostaining protocols to localize several different intracellular, some intranuclear proteins. I was hoping to benefit from both the ease and high sensitivity of the avidin-biotin system and the advantages of recording fluorescent images with a confocal microscope.
The tissue is frozen sections of paraformaldehyde fixed mouse retina. Following a blocking step, I used 2 different antibodies, one a mouse monoclonal, the other, a rabbit polyclonal. After overnight incubation with the primary antibodies, both at R.T. and 4 deg., I applied biotinylated mouse/rabbit IgG, followed by fluorescein conjugated streptavidin. To my greatest diasppointment, all I saw was a high uniformly fluorescent background throughout the entire retina for both antibodies.
Am I leaving out a critical step?
Any suggestions would be highly appreciated.
Judy Trogadis Eye Research Institute of Canada and University of Toronto ph: 416-603-5088 fax: 416-603-5126 email: judy-at-playfair.utoronto.ca
Do any of the universities, companies, hospitals or research institutions have any policies regarding the use of electron microscopy by pregant employees? Is there any literature available on the topic?
Is the advice not to use EM at all during the 9 months, or maybe only during critical periods of development?
In searching the archives I only found one reference on this topic.
Any help would appreciated.
Patrick Diehl Materials Science Department University of North Texas
Dear Patrick, I have sheparded three pregnancies (users, not me) on my 200 kV TEM and I found it a good way to get the instument monitored and thoroughly tested. The last one wore a accumulation monitor at all times and found a curious phenomenon: she accumulated more radiation when sitting at her desk, next to the cinder-block wall in the next room, than when she was working at the TEM at 200 kV. These instruments, when assembled and used properly and monitored in case of leaks after maintenance, leak less x-rays than a TV. I believe the regulations regarding radiation exposure are considered valid for pregnant women.
You wrote:
} Do any of the universities, companies, hospitals or research institutions } have any policies regarding the use of electron microscopy by pregant } employees? Is there any literature available on the topic? } } Is the advice not to use EM at all during the 9 months, or maybe only } during critical periods of development? } } In searching the archives I only found one reference on this topic. } } Any help would appreciated. } } Patrick Diehl } Materials Science Department } University of North Texas } Regards, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
Dear Patrick, I have sheparded three pregnancies (users, not me) on my 200 kV TEM and I found it a good way to get the instument monitored and thoroughly tested. The last one wore a accumulation monitor at all times and found a curious phenomenon: she accumulated more radiation when sitting at her desk, next to the cinder-block wall in the next room, than when she was working at the TEM at 200 kV. These instruments, when assembled and used properly and monitored in case of leaks after maintenance, leak less x-rays than a TV. I believe the regulations regarding radiation exposure are considered valid for pregnant women.
You wrote:
} Do any of the universities, companies, hospitals or research institutions } have any policies regarding the use of electron microscopy by pregant } employees? Is there any literature available on the topic? } } Is the advice not to use EM at all during the 9 months, or maybe only } during critical periods of development? } } In searching the archives I only found one reference on this topic. } } Any help would appreciated. } } Patrick Diehl } Materials Science Department } University of North Texas } Regards, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
I am resending this message to the group becuase of an "undleiverable" message from the orginator's mail system.
HJC
} ---------- } From: Crossman, Harold } Sent: Tuesday, May 13, 1997 3:26PM } To: 'timet-htl' } Subject: RE: Elemental Standards } } We use standards from: } Geller Microanalytical Labs } 426e Boston St. (Rt. 1), Topsfield, MA 01983-1212 } 508 887-7000, fax 508 887-6671, sales-at-gellermicro.com } Website: http:/www.gellermicro.com } } Geller has a varity of elements and will custom fabricate standards if they } can. } You might also want to consider their magnification standard as well. We use } it for both optical and electron scopes. You may be surprised at the } differences between what the scopes reads/images, and what is really there! } } I have no financial interest in promoting Geller. My company has been a } satisfied customer for years. } } } ------------------------------------------------ } Opinions or statements expressed herein, rational or otherwise, do not } necessarily reflect those of my employer. } } Harold J. Crossman } OSRAM SYLVANIA INC. } Lighting Research Center } 71 Cherry Hill Dr. } Beverly, MA 01915 } Phone: (508) 750-1717 } E-mail: crossman-at-osi.sylvania.com } } Our web sites: www.sylvania.com } www.siemens.com } -- } } "Crossman, Harold" {crossman-at-osi.SYLVANIA.com} }
Dear Patrick, I have got two healthy daughters (the youngest one is 10 weeks). During the whole period of both my pregnancies I have worked with electron microscopy (TEM, SEM, low vacuum SEM and environmental SEM). Risoe National Laboratory is a former Atomic Energy Commission Research Establishment, so we still have health physicists around. They were consulted as soon as I realised that I was pregnant. The health physicist measured the radiation level at all our TEMs and SEMs. Only the background level of approx. 0.06 microSv/h was found. During the my pregnancies I wore two badges that checked the radiation continuosly. Nothing above background level was detected.
Charlotte C. Appel Materials Research Department Risoe National Laboratory DK-4000 Roskilde DENMARK radiation
It sounds like the problem may be using a monoclonal mouse antibody on mouse tissue. When you come in with your secondary against mouse IgG it will find a whole bunch of it in the tissue. If you don't have any choice but to use a monoclonal on the mouse tissue you can come in with a 10x excess of anti-mouse fab fragments to block all the mouse IgG, then cone in with your Primary.
Robert Underwood Morphology Core U of Washington
On Tue, 13 May 1997, Judy Trogadis wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Fellow immunostainers, } } I am using immunostaining protocols to localize several different } intracellular, some intranuclear proteins. I was hoping to benefit from } both the ease and high sensitivity of the avidin-biotin system and the } advantages of recording fluorescent images with a confocal microscope. } } The tissue is frozen sections of paraformaldehyde fixed mouse retina. } Following a blocking step, I used 2 different antibodies, one a mouse } monoclonal, the other, a rabbit polyclonal. After overnight incubation } with the primary antibodies, both at R.T. and 4 deg., I applied biotinylated } mouse/rabbit IgG, followed by fluorescein conjugated streptavidin. To my } greatest diasppointment, all I saw was a high uniformly fluorescent } background throughout the entire retina for both antibodies. } } Am I leaving out a critical step? } } Any suggestions would be highly appreciated. } } Judy Trogadis } Eye Research Institute of Canada and } University of Toronto } ph: 416-603-5088 } fax: 416-603-5126 } email: judy-at-playfair.utoronto.ca }
Radiation exposure should be limited to a fetus, particularly during the first 3 months. The Nuclear Reg. Comm. limits this exposure to {500 mRem for the 9 months. This SHOULD be a huge value compared to any exposure from any EM unless it is a HVTEM that is improperly setup (you must stay out of the chamber when running {g} ). A SEM running 30 kV. cannot produce any x-rays over 30 keV - Most are well under. These relatively soft x-rays are (typically) completely shielded by the column and chamber walls....Consult your manufacutrer for detailed exposure rates. I think you will find that a cross country air flight (or 9 months in Denver vs. a sea level city) will expose you to more radiation than will 9 months in front of the EM.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Do any of the universities, companies, hospitals or research institutions have any policies regarding the use of electron microscopy by pregant employees? Is there any literature available on the topic?
Is the advice not to use EM at all during the 9 months, or maybe only during critical periods of development?
In searching the archives I only found one reference on this topic.
Any help would appreciated.
Patrick Diehl Materials Science Department University of North Texas
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Rui:
I assume you are looking for Dr. Charles Garber of Structure Probe/SPI Supplies. I have copies below the signature block from one of his messages.
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI Structure Probe, Inc. FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
David Henriks TEL: 800-728-2233 (toll-free in USA) South Bay Technology, Inc. 714-492-2600 1120 Via Callejon FAX: 714-492-1499 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 Rui Costa }
I am looking for Dr Charles Graber (or Graber). Sorry I do not if the last name is correct.
After a few hours using the TEM (Philips CM30) with the cold trap filled with LN2, it appears that some vapors in the microscope column get trapped, as it should be.
Now the problem is: have you ever noticed any problem AFTER removing the LN2 dewar, thus heating the trap, provoking a degassing in the column. We have noticed that after several hours working with the cold trap, the IGP gas pressure level increases a lot after removing the dewar (it happens that HT switches off during the night because of that IGP pressure increasing above the "safety level"), and some users here tell me that it should not be that way.
Question: did you ever notice such a problem, and if yes, how long time would you work without heating the trap in order not to get any dramatic increase of IGP pressure after removal of the dewar.
} After a few hours using the TEM (Philips CM30) with the cold trap filled } with LN2, it appears that some vapors in the microscope column get } trapped, as it should be. } } Now the problem is: have you ever noticed any problem AFTER removing the } LN2 dewar, thus heating the trap, provoking a degassing in the column. We } have noticed that after several hours working with the cold trap, the IGP } gas pressure level increases a lot after removing the dewar (it happens } that HT switches off during the night because of that IGP pressure } increasing above the "safety level"), and some users here tell me } that it should not be that way. } } Question: did you ever notice such a problem, and if yes, how long time } would you work without heating the trap in order not to get any dramatic } increase of IGP pressure after removal of the dewar.
Yes, we notice this on both our Philips CM-12 and CM-20. Since our lab does a lot of cryoTEM, it would be especially bad if we did not deal with the problem. At the end of each day we turn off the high tension and then use the Philips CRYO function to turn off the ion getter pump and open V6 so that the diffusion pump pumps the column. We then replace the cold-trap Dewar vessel with a vessel filled with warm water and permit the diffusion pump to pump away all the contaminants (it takes about 30 min.) We then switch off the CRYO function, which turns the ion getter pump back on. After about another 15-30 min when the vacuum has recovered, we switch on the HT (on conditioning) and leave the microscope for the night. We get very little downtime by following this procedure. I should point out that the power in Rochester is very reliable, so we have only had one unplanned power outage in the last five years. Under these conditions, the microscope seems to perform best when the high tension is left on continuously. -- Best Regards, John Minter
Eastman Kodak Company Phone: (716) 722-3407 Analytical Technology Division FAX: (716) 477-3029 Room 2112 Bldg 49 Kodak Park Site email: minter-at-kodak.com Rochester, NY 14562-3712 calendar: via PROFS
} Do any of the universities, companies, hospitals or research institutions } have any policies regarding the use of electron microscopy by pregant } employees? Is there any literature available on the topic? } } Is the advice not to use EM at all during the 9 months, or maybe only } during critical periods of development? } } In searching the archives I only found one reference on this topic. } } Any help would appreciated.
Dear Patrick,
Everyone who uses radiation-producing equipment should be monitored. We have ion-chamber dosimeters for occasional users and film badges for people who are around the instrument every day. The limit for exposure for pregnant women is 500 millirem over the course of the pregnancy, but this amount should not happen in a short period. There is an unofficial limit of 50 millirem per month. These numbers come from the experts in the NY State Department of Health. For exposures to x-rays, rad and rem are equi- valent, and 1 rad (Radiation Absorbed Dose) is 100 ergs per gram. If you have no capability for monitoring users, the next best thing is to use a detector, such as a Geiger counter or an ion chamber, to deter- mine whether the radiation near the microscope is different from background. If there is no difference between the radiation measured before the scope is turned on and when it is cranked up fully, then you can be reasonably sure there is no danger (assuming you have a detector which is sensitive to the radiation your scope produces); however, this probably won't satisfy any legal requirements. Yours, Bill Tivol
There is no danger from contemporary microscopes which are well maintained and monitored. The problem exists with chemicals. Cacodylate buffer if used in a laboratory eventually contaminates bottles, hoods, etc., with fine arsenic dust. Then there is the lead stain and the UA. Anyone pregnant should not be weighing out any chemical containing heavy metals. Extreme care needs to be taken with other noxious things such as glut, paraform, xylene, propylene, etc. During my time in another lab, there occurred a chemical spill - the holding tank backed up into the lab. A mixture of the above seeped up through the floor drains. My coworker attempted to contain the spill with a properly fitted mask. At the time she was 12 days pregnant. At seven months she gave birth to a badly deformed baby which lived only for an hour. After exhaustive investigation, it was decided that the uterine fluid had been contaminated with heavy metals. The enormous quantity of fluid gave her the appearance at seven months of a 10 months preganancy, at least. Anyone who is not protected by birth control and might be pregnant at any time should not be weighing out questionable material, and should take extreme care with being gloved, etc., at all times. So long, Hildy
In message {Pine.LNX.3.93.970514184137.22551B-100000-at-giga.sct.ub.es} Yves Maniette writes: } } After a few hours using the TEM (Philips CM30) with the cold trap filled } with LN2, it appears that some vapors in the microscope column get } trapped, as it should be. } } Now the problem is: have you ever noticed any problem AFTER removing the } LN2 dewar, thus heating the trap, provoking a degassing in the column. We } have noticed that after several hours working with the cold trap, the IGP } gas pressure level increases a lot after removing the dewar (it happens } that HT switches off during the night because of that IGP pressure } increasing above the "safety level"), and some users here tell me } that it should not be that way. } } Question: did you ever notice such a problem, and if yes, how long time } would you work without heating the trap in order not to get any dramatic } increase of IGP pressure after removal of the dewar. } } Yves MANIETTE } Universitat de Barcelona
Yves, Yes I see this problem once and awhile with our CM-12. Seems to occur if we've had samples in the column that are volatile and thus the cold trap collects more junk, which can be released suddenly into the column when the cold trap warms up.
Two thoughts: 1. Instead of removing the dewar, leave it on and allow the liquid nitrogen to evaporate with the dewar installed. I think the warm-up rate would be reduced, compared to removing the dewar and exposing the copper braid to room air, such that the IGP and other pumps may be able to handle the gas load without crashing the vacuum system.
2. But if you still get crashes, turn off the IGP until the diffusion pump has time to recover the system. I had my service engineer install a simple toggle switch on the back side of the column cover to allow me to shut off the IGP, as my instrument did not come configured with any software switch to do this.
Typically, in my lab, a crash would occur late afternoon after my last TEM user has left and the cold trap warms up. Then I shut off the IGP for overnight and turn it on the next morning.
Hope this helps.
Gib
--
Gib Ahlstrand, MMS Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
Mn Microscopy Society Home page: http://charfacnu.cie.umn.edu/MnMicSoc.html
Microscopy & Imaging Consortium Home page: http://biosci.umn.edu/MIC/consortium.html
DEPARTMENT OF PHYSICS, UNIVERSITY OF ILLINOIS AT CHICAGO
Two postdoctoral positions are available in the Interface Physics Group at the University of Illinois at Chicago (UIC). Research in the Interface Physics Group focuses on the use atomic resolution imaging and analytical techniques in electron microscopy, coupled with theoretical simulations, to determine the structure-property relationships at internal interfaces on the fundamental atomic scale. Current research programs involve ceramics, high-Tc superconductors and optoelectronic/high-power semiconducting materials and devices. The experimental facilities to perform this research are comprehensive: a JEOL 2010 Field-Emission STEM/TEM featuring a 1.4=C5 probe size, "drift free" stage, high-angle annular dark-field detecto= r (Z-contrast), Gatan Imaging Filter, and Noran EDS; a VG HB501A =46ield-Emission dedicated STEM with EDS, EELS and Auger spectrometers; a JEOL 3010 conventional TEM with digital imaging capabilities and EDS; a JEOL 6320 Field-Emission SEM with EDS and Cathodoluminescence; a JEOL JXA733 microprobe; and a Topometrix AFM/STM. In addition to the electron microscopes, specimen preparation facilities include a Gatan Duo-mill, =46ischione precision ion-mill, Fischione plasma cleaner and Leica Ultramicrotome. The Interface Physics Group has a Silicon Graphics R10000 Power Indigo workstation with a Molecular Simulations' Cerius 2 package incorporating the CASTEP pseudopotential code. The physics department has additional workstations and access to the UIC Convex Exemplar Supercomputer and the National Center for Supercomputing Applications at UIUC. The two research positions are as follows:
(I) Atomic Resolution Imaging and Analysis Techniques
With the demise of VG as a supplier of state-of-the-art analytical electron microscopes, it is essential that atomic resolution imaging (Z-contrast) and analysis techniques (EELS and EDS) are developed on other instruments. The JEOL 2010F has the potential to exceed both the imaging and analytical performance of the VG instruments, with a resolution at 200kV of 1.4=C5. Through an NSF award, UIC has recently purchased this microscope with the primary aim of developing the next generation of high resolution imaging/analytical microscope. Coupled with the purchase of this microscope is a postdoctoral position to perform the initial instrument characterization and coordinate its application to research programs at UIC and at neighboring institutions (Argonne National Laboratory, Northwestern University and Illinois Institute of Technology). This position therefore offers the potential to develop a wide range of research programs, in addition to a unique opportunity to address fundamental issues concerning incoherent vs coherent imaging and microanalysis. The postdoctoral position will also have primary responsibility for the VG HB501A STEM and JEOL 3010 TEM. At the end of the 2-year funding by NSF for this position, UIC will be seeking a permanent electron microscope facility manager.
(II) Grain Boundaries in Ceramics
As part of a DOE funded program, a postdoctoral position is available for the investigation of the structure-property relationships at grain boundaries in ceramics. The aim of this program is to correlate the experimental results from atomic resolution imaging and microanalysis to produce structural models which can be compared with theoretical simulations. Of particular interest is the use of EELS to characterize the 3-dimensional structure of the grain boundary and quantify the number of vacancies and dopant atoms present in the structure. This information can only be obtained through the use of EELS. This program necessarily includes the use of multiple scattering analysis of the EELS fine-structure, the use of bond-valence sum analysis to produce preliminary models and the application of the CASTEP simulation codes for detailed analysis of the boundary structure. Prior experience in these techniques is not essential, only a willingness to learn how to use the commercially available software packages.
Successful candidates will be recent Ph.D. graduates in physics, metallurgy, or materials science with a sound background in the relevent materials issues and an ambition to be part of a developing program pushing at the frontiers of interface physics. Please send a resume and publication list to Professor N. D. Browning at the address below. Prior experience in STEM or TEM is essential. However, consideration will be based on the candidates overall potential for success in the field and applicants with prior experience in related fields are encouraged to apply. Positions are for one year initially, normally renewed for a second year with possibilities existing for further years. Salary is commensurate with experience. UIC is an equal opportunity employer.
*************************************************************** Nigel D. Browning PhD, Assistant Professor, Interface Physics Group, Department of Physics (M/C 273), University of Illinois at Chicago, 845 West Taylor Street, Chicago. Il 60607-7059
} The last one wore a accumulation monitor at all times and found a curious } phenomenon: she accumulated more radiation when sitting at her desk, next to } the cinder-block wall in the next room, than when she was working at the TEM } at 200 kV.
Dear Mary, This is not so curious. The potassium-40, naturally occurring in concrete is a well-known source of radiation. If the user had her desk in Denver, there would have been an additional component from the greater cosmic ray flux at high altitude. Yours, Bill Tivol
You may consider to put in the specimen and let it pump for about 15 min. before filling the cold trap. } All, } } After a few hours using the TEM (Philips CM30) with the cold trap filled } with LN2, it appears that some vapors in the microscope column get } trapped, as it should be. } } Now the problem is: have you ever noticed any problem AFTER removing the } LN2 dewar, thus heating the trap, provoking a degassing in the column. We } have noticed that after several hours working with the cold trap, the IGP } gas pressure level increases a lot after removing the dewar (it happens } that HT switches off during the night because of that IGP pressure } increasing above the "safety level"), and some users here tell me } that it should not be that way. } } Question: did you ever notice such a problem, and if yes, how long time } would you work without heating the trap in order not to get any dramatic } increase of IGP pressure after removal of the dewar. } } Yves MANIETTE } Universitat de Barcelona
Ke Han Center for Materials Science Los Alamos National Laborotory Los Alamos, New Mexico NM87545, USA Tel: 505-6650771 Fax: 505-6652992
We see exactly the same phenomenon on two separate TEM's (Hitachi H500 and H7100). After an initial improvement in vacuum due to application on the LN2 on the specimen cold trap (but oddly, not the DP traps), we get good performance for 2-3 hr. Then, after the LN2 level goes below the transfer probe or braid, the vacuum deteriorates to the point that the HT kicks off. Very predictable: so we simply keep adding LN2 after 2 hr until the HT is turned off for the evening. Now, if one does not add the LN2 to the specimen trap to begin with, the phenomenon never happens. So, I conclude that the loss of HT must be due to the sudden release of a lot of contamination (water vapor, organics from the specimen). We do not see this with the DP traps because the vapors must be removed by the DPs. Wil Bigelow probably knows what's going on here, so I will wait for his comments.
} After a few hours using the TEM (Philips CM30) with the cold trap filled } with LN2, it appears that some vapors in the microscope column get } trapped, as it should be. } } Now the problem is: have you ever noticed any problem AFTER removing the } LN2 dewar, thus heating the trap, provoking a degassing in the column. We } have noticed that after several hours working with the cold trap, the IGP } gas pressure level increases a lot after removing the dewar (it happens } that HT switches off during the night because of that IGP pressure } increasing above the "safety level"), and some users here tell me } that it should not be that way. } } Question: did you ever notice such a problem, and if yes, how long time } would you work without heating the trap in order not to get any dramatic } increase of IGP pressure after removal of the dewar. } } Yves MANIETTE } Universitat de Barcelona
John Bozzola 71 Concordia Drive Carbondale, IL 62901 Internet address: JBozzola-at-aol.com
------------------------------------- I need information on linking the video signal of a SEM (1V p-p) composite video single to a LECO imagin system that can accept a variety of formats (PAL, NTSC) etc. Any ideas or experience with this? Thanks Craig
Hi all I have a question for all the "digital microscopysts: I need to mount a ccd system to my OLYMPUS BX 50 microscope. I want to do optical sectioning and fluorescence imaging. Some people tell my that the more convenient is a digital ccd camera B&W, whit a resolution of 758 x 512 or 1024 x 1024. What is your opinion of these points?. thanks in advance =================================================== Fernando D. Balducci Laboratorio de Microscopia Electronica Facultad de Ingenieria - Bioingenieria Universidad Nacional de Entre Rios Argentina e-mail: microsc-at-fi.uner.edu.ar tel: 54 43 975100 fax: 54 43 975077 ===================================================
In reply to Yves Maniette query on cold trap degassing:
Philips told me to keep the HT of my CM20 on all the time. Unfortunately, when the cold trap warms up at the end of the day, off goes the HT! Of course, the more samples going in and out the microscope during the day, the further the IGP(ion getter pump) reading goes up as one would expect. I normally take off the cold trap dewar when I have finished for the day since I wish to reduce the nitrogen bill - I put the nitrogen in the ion mill dewar.
I also would be interested to know the best way to tackle the problem. So far all I have got is shrugs!
I also think that there should be a toggle switch to take the ion getter off, removing and replacing the back panel is an unnecessary pain. Mike
Michael J Witcomb PhD Electron Microscope Unit University of the Witwatersrand Private Bag 3 WITS 2050 South Africa
} In reply to Yves Maniette query on cold trap degassing: .../... } } I also think that there should be a toggle switch to take the ion } getter off, removing and replacing the back panel is an unnecessary } pain. } Mike
You can ask your local maintenance engineer to place a switch on the microscope SIDE panel near the IGP pump. This switch shoult be connected to the small socket down the IGP pump. Any Philips Engineer also knows that trick and will install it for you if you ask him. The only point is NOT to place the switch on the back panel, because that will be a hindrance when you really want to remove that panel...
Microscopy-at-sparc5.microscopy.com Original-Encoded-Information-Types: IA5-Text X400-Content-Type: P2-1984 Message-ID: {"ISOPRO::DH-EF::5080::337B189B"*/G=Allen/S=White/O=txmta1/PRMD=AMD/ADMD=ATTMAIL/C=US-at-MHS} Importance: normal
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have noticed such a problem with a CM30 at a previous worksite. It was first evident after installation of a new filament which had an asymmetrical emission pattern within a couple of days of operation. I hooked a dual channel chart recorder to X10, the ion pump current monitor test point, and to the output of a nude ion gage installed near the column vacuumn system. After leaving the cold trap unattended at 5PM I found on monitoring the chart recorder that the cold trap would regenerate during the night, early hours of morning, with concommitent increase of ion pump current and ion guage voltage. Depending on the number of sample exchanges made during the day the column backing pipe pressure would go as high as 1E-5 Torr according to to the ion guage.
By my experience you should an have at least an hour or so of use after removing the LN2 before the cold trap begins to warm up. Unfortunately, the CM30 has a filament preheat voltage that remains after the filament knob is turned all the way down and is not interrupted until the IGP interlock circuit cuts off the high voltage. If you experience high tension trips frequently you may have a vacuum leak and the filament may die prematurely if it is LaB6.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
All,
After a few hours using the TEM (Philips CM30) with the cold trap filled with LN2, it appears that some vapors in the microscope column get trapped, as it should be.
Now the problem is: have you ever noticed any problem AFTER removing the LN2 dewar, thus heating the trap, provoking a degassing in the column. We have noticed that after several hours working with the cold trap, the IGP gas pressure level increases a lot after removing the dewar (it happens that HT switches off during the night because of that IGP pressure increasing above the "safety level"), and some users here tell me that it should not be that way.
Question: did you ever notice such a problem, and if yes, how long time would you work without heating the trap in order not to get any dramatic increase of IGP pressure after removal of the dewar.
In order to answer this accurately, need to know a bit more about the SEM video than it's amplitude. Is it NTSC? Slow Scan? The 1 volt p-p video is a standard "line" level.... Woody
------------------------------------- I need information on linking the video signal of a SEM (1V p-p) composite video single to a LECO imagin system that can accept a variety of formats (PAL, NTSC) etc. Any ideas or experience with this? Thanks Craig
On May 14, 6:51pm, Yves Maniette wrote: } Subject: TEM Cold trap degassing } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } All, } } After a few hours using the TEM (Philips CM30) with the cold trap filled } with LN2, it appears that some vapors in the microscope column get } trapped, as it should be. } } Now the problem is: have you ever noticed any problem AFTER removing the } LN2 dewar, thus heating the trap, provoking a degassing in the column. We } have noticed that after several hours working with the cold trap, the IGP } gas pressure level increases a lot after removing the dewar (it happens } that HT switches off during the night because of that IGP pressure } increasing above the "safety level"), and some users here tell me } that it should not be that way. } } Question: did you ever notice such a problem, and if yes, how long time } would you work without heating the trap in order not to get any dramatic } increase of IGP pressure after removal of the dewar. } } Yves MANIETTE } Universitat de Barcelona } -- End of excerpt from Yves Maniette
It is same with our CM300FEG and CM120, especially after a few hours cryo work or change sample several times. But I think this is normal. As far as the dewar is filled with LN2, you can work as long as you want. With our CM300FEG, there is an extra valve V7 which should be manually closed after finishing the work. Soon after you remove the dewar or run out the LN2, the IGP1 will increase quite a lot (} 19, which is the safe value for open the V7), and then come down below 19 after a while. If V7 is closed, there should be no harm to the TEM. If you have the cryo option in the vacumm page, you can also turn off the IGP1 for a while (I usually turn it off for half hour after remove the dewar), and let ODP to pump the column, which is more efficient to remove the water vapor from the column. But this works only when your P3 is very good, definitely before changing film.
Yifan Cheng
--
************************************************************************** * Dr. Yifan Cheng * Phone: +1-904-644-4104 * * Institute of Molecular Biophysics * Fax: +1-904-561-1406 * * Florida State University * Email: ycheng-at-sb.fsu.edu * * Tallahassee, FL 32306-3015, U.S.A. * http://www.sb.fsu.edu/~ycheng * *------------------------------------------------------------------------* * Home address: * * * 313 Pennell Circle #4 * Phone: +1-904-575-3620 * * Alumni Village * * * Tallahassee, FL32310, U.S.A. * * **************************************************************************
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Pregnancy in EM 5/15/97 7:54 AM
I was wondering what most EM labs do for precautionary measures when someone in the lab is pregnant. I have a colleague who is pregnant now, and she does not do resin embedding. She does cut cured blocks,stain them with uranyl acetate and lead citrate, and look at them on the TEM. Is this normal procedure in other labs? Should she wear a mask when staining? Linda Chicoine Center for Cell Imaging Yale University New Haven, CT
Greetings, I wonder anyone knows of a source for Technovit resin in the USA? This resin is made by Hareus (spelling?) in Europe. Is there perhaps a similar if not identical resin available here? Also, does anyone know if Technovit is glycol methacrylate? Thanks in advance for any leads, Tobias Baskin
On the Philips microscopes there is a socket in the lower part of the IGP. if this socket is disconnected then IGP will be disconnected, and also the vacuum valves will open in such a way that the ODP (low chamber) will be more openly connected with the column and gun. This allows to help "cleaning" the IGP gun.
But the socket is hidden behin the rear panel so that you have to remove it in order to remove the socket, something not very practical. The hint is to connect the socket to a switch that you place at any convenient place, and the side panel is a convenient one.
On Thu, 15 May 1997, Ann-Fook Yang wrote:
} I follow the thread with interest. } What is the switch, you talked about, for? } When do you use it? } Just curious. } } Ann Fook } } } } } } Yves Maniette {yves-at-giga.sct.ub.es} 05/15/97 06:18am } } } } ------------------------------------------------------------------------ } } You can ask your local maintenance engineer to place a switch on the } microscope SIDE panel near the IGP pump. This switch shoult be } connected } to the small socket down the IGP pump. Any Philips Engineer also knows } that trick and will install it for you if you ask him. The only point is } NOT to place the switch on the back panel, because that will be a } hindrance when you really want to remove that panel... } } Yves MANIETTE } Universitat de Barcelona } }
Yves MANIETTE Universitat de Barcelona Serveis Cientifico Tecnics Unitat ESCA TEM Carrer Lluis Sole i Sabaris, 1-3 E-08028 BARCELONA ESPANYA
I am interested in purchasing a set of ZEISS Plan Apo objectives for my Photomicroscope II.
These must be 160mm tube length and be in mint condition. I need the following: 4X, 10X, 20X, 40X, 63X and 100X (the 63 and 100X are O.I.). Will consider other Zeiss types but prefer Plan Apo.
Will also consider purchasing a complete vintage (1975/1990) Zeiss scope.
Thanks for any leads that may result in a favorable purchase.
Cordially, Gil Groehn ULTRAMED, INC. - RESEARCH DIV.
-- ============================================================= ULTRAMED, INC. Research Div. ad408-at-detroit.freenet.org Grosse Pointe Farms, MICH 48236 USA PHONE: 313-884-1139 ===============================================================
Electron Microscopy Technician - EM technician needed by state-of-the-art laboratory studying the organization of proteins and nucleic acids in cell nuclei. The individual should be highly competent in thin sectioning with a diamond knife, fixation and embedding methods, darkroom work, immunofluorescence microscopy and/or fluorescence in situ hybridization. Experience in cell culture and immunogold labeling is highly desirable. Please send resume and two letters of reference to: Dr. David L. Spector, Cold Spring Harbor Laboratory, P.O. Box 100, Cold Spring Harbor, New York 11724.
} Appello Urgentissimo - VERY URGENT APPEAL } } Urge aiuto per una bambina italiana di sei anni Francesca De Lunas } FRANCESCA DE LUNAS IS A 6 YRS OLD DAUGHTER WHO URGENTLY NEEDS HELP. } } ricoverata presso il reparto rianimazione dell'ospedale Brotzu di } Cagliari } (Sardegna-Italia) Dott. Pettinao. } SHE'S NOW UNDER THERAPY BY DR. PETTINAO AT THE "REPARTO RIANIMAZIONE" } [REANIMATION DEPT.], } OSPEDALE BROTZU, CAGLIARI (SARDINIA, ITALY) } } La bambina presenta una sindrome caratterizzata da: } SHE SHOWS A SYNDROME WHICH PECULIARITIES ARE: } } - ipercoagulazione del sangue; } BLOOD HYPERCOAGULATION } } - trombi che si formano in circa 15 secondi di colore rosso con nucleo } bianco } di lunghezza di circa 2-3 cm. } RED THROMBI WITH WHITE NUCLEUS, ABOUT 2-3 CM LONG, WHICH FORM IN 15 } SECS. } } Alla bambina e' stata gia' amputata la gamba sinistra e, finora non e' } stato possibile effettuare diagnosi di alcun tipo. } THE DAUGHTER HAS ALREADY BEEN AMPUTATED THE LEFT LEG. IT HAS NOT BEEN } POSSIBLE } TO MAKE ANY DIAGNOSIS UNTIL NOW. } } Chiunque sia in grado di ipotizzare una diagnosi o di riconoscerla } si metta urgentemente in contatto con le seguenti e-mail: } ANYBODY ABLE TO MAKE A DIAGNOSIS OR RECOGNIZE THIS DISEASE PLEASE GET } IN } TOUCH } URGENTLY BY E-MAIL TO THE FOLLOWING ADDRESSES: } } lukrezia-at-mbox.vol.it } } stefania-at-alanet.it } } schintu-at-pan.bio.uniroma1.it } } psico-at-mbox.vol.it } ********************************* } THANK YOU VERY MUCH FOR YOUR COOPERATION ____________________________________________________________________________ ____
Dr. Cristiano Rumio Istitute of Human Anatomy Via Mangiagalli 31 20133 Milan Italy Tel. -39-2-2663683 Fax. -39-2-2364082 E-mail: crylsm-at-imiucca.csi.unimi.it URL: http://imiucca.csi.unimi.it/~endomi/confocal.html Immunofluorescence Course http://imiucca.csi.unimi.it/endomi/ACIF.html ____________________________________________________________________________ ____
Appello Urgentissimo - VERY URGENT APPEAL } } Urge aiuto per una bambina italiana di sei anni Francesca De Lunas } FRANCESCA DE LUNAS IS A 6 YRS OLD DAUGHTER WHO URGENTLY NEEDS HELP. } } ricoverata presso il reparto rianimazione dell'ospedale Brotzu di } Cagliari } (Sardegna-Italia) Dott. Pettinao. } SHE'S NOW UNDER THERAPY BY DR. PETTINAO AT THE "REPARTO RIANIMAZIONE" } [REANIMATION DEPT.], } OSPEDALE BROTZU, CAGLIARI (SARDINIA, ITALY) } } La bambina presenta una sindrome caratterizzata da: } SHE SHOWS A SYNDROME WHICH PECULIARITIES ARE: } } - ipercoagulazione del sangue; } BLOOD HYPERCOAGULATION } } - trombi che si formano in circa 15 secondi di colore rosso con nucleo } bianco } di lunghezza di circa 2-3 cm. } RED THROMBI WITH WHITE NUCLEUS, ABOUT 2-3 CM LONG, WHICH FORM IN 15 } SECS. } } Alla bambina e' stata gia' amputata la gamba sinistra e, finora non e' } stato possibile effettuare diagnosi di alcun tipo. } THE DAUGHTER HAS ALREADY BEEN AMPUTATED THE LEFT LEG. IT HAS NOT BEEN } POSSIBLE } TO MAKE ANY DIAGNOSIS UNTIL NOW. } } Chiunque sia in grado di ipotizzare una diagnosi o di riconoscerla } si metta urgentemente in contatto con le seguenti e-mail: } ANYBODY ABLE TO MAKE A DIAGNOSIS OR RECOGNIZE THIS DISEASE PLEASE GET } IN } TOUCH } URGENTLY BY E-MAIL TO THE FOLLOWING ADDRESSES: } } lukrezia-at-mbox.vol.it } } stefania-at-alanet.it } } schintu-at-pan.bio.uniroma1.it } } psico-at-mbox.vol.it } ********************************* } THANK YOU VERY MUCH FOR YOUR COOPERATION } send to your mailing list
Hello: Anybody who has the experience in using liquid helium stage for either TEM or SEM, or has the information on where I can find this type of stage, please send me an email at wtao-at-mtu.edu. Thanks.
A colleague is interested in purchasing a used, inexpensive light microscope. Nothing fancy: standard LM with 10x, 40x and 100x objectives and reasonable resolution. To be used for examination of biological and/or clinical materials (smears, sections, etc). Private individuals or vendors should contact:
Contact: Dr. Faiqa Hassan St. Louis, Mo. Phone: 314-522-0083 Fax: 314-322-0083
At 10:22 AM 5/15/97 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have a small business which deals mostly with electron optics but when ever I have needed help with glass optics, I contact Mel Sobel Microscopes. Their address is 29 Louis Street, Hicksville, New York 11801 Phone: 516/935-7794 FAX 516/935-6131. These people seem to have parts for any optical microscope ever made and their prices are very reasonable.
Good luck.
I am not associated with Mel Sobel Microscopes. } } } } } I am interested in purchasing a set of ZEISS } Plan Apo objectives for my Photomicroscope II. } } These must be 160mm tube length and be in mint } condition. I need the following: 4X, 10X, 20X, } 40X, 63X and 100X (the 63 and 100X are O.I.). } Will consider other Zeiss types but prefer Plan Apo. } } Will also consider purchasing a complete vintage } (1975/1990) Zeiss scope. } } Thanks for any leads that may result in a favorable } purchase. } } Cordially, } Gil Groehn } ULTRAMED, INC. - RESEARCH DIV. } } -- } ============================================================= } ULTRAMED, INC. Research Div. ad408-at-detroit.freenet.org } Grosse Pointe Farms, MICH 48236 USA PHONE: 313-884-1139 } =============================================================== } } ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ Alexander Greene Scientific Instrumentation Services, Inc. Number 499, Post Office Box 19400 Austin, Texas 78760 Phone: 512/282-5507 FAX 512/280-0702
REASONABLY PRICED ELECTRON MICROSCOPE REPAIR ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
Microscopy and Microanalysis '97 is less than three months away. The meeting this year will be held August 10-14 in Cleveland, Ohio. The deadline for early registrations is July 15. If you are plan to attend, please consider making plans as soon as possible. Cleveland has recently become a popular place to visit and hotel rooms WILL be at a premium if you don't act soon. A popular attraction, the Cleveland Indians, will be playing at home that week. The baseball games and other attractions such as the Rock and Roll Hall of Fame have already lead to a huge demand for hotel space for that week.
Information on the Meeting proceedings can be found on the world wide web at http://www.bright.net/~strecker/msno/mm97.html. The forms you will need for early meeting registration and hotel registration are available in pdf format for downloading here. You will also find information on hotels, events and local attractions available to attendees of the meeting at this site. If you require additional information or are unable to access the WWW, you can contact the MSA business office by e-mail at BusinessOffice-at-MSA.Microscopy.com or by phone at (800) 538-3672.
Once again, please register early so you dont miss out on Microscopy and Microanalysis 97. I hope to see you in Cleveland this year.
A colleague studying trace metals in San Francisco Bay has noticed more osmium in the sediments than expected.
He knows EM-types use osmium and quizzed me regarding possible sources for the Os in the bay. He wondered how many EM labs are around the bay and if they could be sources of Os in the waste stream. I said probably not, most EM-types are careful about disposing of all toxic/hazardous waste in an approved manner. He was still curious so I offered to pose a few questions to the group on his behalf.
If you have any suggestions or information to add to his research please send them my way. Here are some of his specific questions:
1. How many EM labs that use OsO4 are there around the SF Bay area?
2. About how much OsO4 do they use in a year?
3. How do the labs dispose of waste OsO4 and does that include every last bit, even that from rinsing used containers?
4. If the Os he sees is not from EM labs, then where could it be coming from? Does anyone know of industrial or other applications that use a lot of Os?
Stay tuned for further details as he develops his theories.
Thanks,
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
A colleague has asked me to post this request. He is seeking information on short courses which may be available in confocal laser scanning microscopy (general principles, applications, etc. in the biological and biomedical sciences). The course(s) should preferably be offered in the continental U.S. The first choice would be in the Rocky Mountain or Desert Southwest areas.
I've perused the (published) lists of the Lehigh and Woods Hole courses, and nothing jumps out at me as addressing specifically confocal laser microscopy. Perhaps I've missed something??
Does anyone have a recommendation as to who my colleague should contact? If so, please e-mail me directly. I will pass the info along.
BTW, it might also be worth posting your reply to the Microscopy Listserver. If there is such a course coming up this year there may be others who would be interested.
it's glycol methacrylate and is the same stuff as Historesin under a different name.
At 08:42 AM 5/15/97 -0600, Tobias Baskin wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Delilah Wood United States Department of Agriculture Western Regional Research Center 800 Buchanan Street Albany, CA 94710
I would like to make contact with those of you who use beryllium grids fairly often or occasionally in your work, for the purpose of discussing some particular issues with those grids. I am concerned, really, about the toxicity of that material. Are there any viable alternatives? Or is the hazard, to operator or environment, in fact not so worrisome that anyone really need be concerned about looking for something else? (Opinions?) I'm also interested to learn for what work it is generally felt that one should even be considering a beryllium grid, since they are pretty costly to boot!
Cheers,
Cam
____________________________________________________________________________ P.O. Box 1932 Main Station T: 1 403 913 3850 Cameron Sorlie, President Edmonton, AB T5J 2P3 Canada F: 1 403 433 9376 General Microdevices, Inc. ----------------------------------------------- generalmicro-at-incentre.net ______________________________________________________ Microfabrication technology applications
} I would like to make contact with those of you who use beryllium grids } fairly often or occasionally in your work, for the purpose of discussing } some particular issues with those grids. I am concerned, really, about the } toxicity of that material. Are there any viable alternatives? Or is the } hazard, to operator or environment, in fact not so worrisome that anyone } really need be concerned about looking for something else? (Opinions?) I'm } also interested to learn for what work it is generally felt that one should } even be considering a beryllium grid, since they are pretty costly to boot! } } } Cheers, } } Cam
Beryllium grids can be very useful when doing X-ray microanalysis - especially if you are looking for Cu, or anything with a peak that might be swamped by Cu. Also potentially useful if you are looking for light elements whose lines might be swamped by the Cu L line.
There are alternatives - Al grids resolve most of the above problems. Carbon coated nylon and carbon composite grids are also available from the usual suppliers - Agar, SPI, etc. However, for really critical analysis, I'd say Be grids are to be preferred.
I don't believe there is any serious hazard associated with these grids. Be is a problem if it get into the lungs (or trapped under the skin), where it leads to medical conditions similar to those caused by asbestos. So don't use abrasives on it, and I guess you want to avoid exposure to any strong acids that might attack the oxide surface (and don't eat them:)
a new tipe of SPM tip artifact is described and put on the WWWW:
http://newton.phy.bme.hu/pub/corfu93/index.html
may be it is interesting for some of you!
Best regards: Peter
*************************************************************************** * dr. Nagy, Peter (physicist) * * MTA (Hungarian Academy of Sciences) * * KFKI (Central Research Institute for Physics * * ATKI (Research Institute for Materials Sciences) * * STM (Scanning Tunneling Microscopy Group) * * H-1525 Budapest-114, P.O.Box.49 Tel.: +36-1-3-959-220/19-68 * * Budapest, Konkoly T.M. u. 29-33 Fax.: +36-1-3-959-284 * ***************************************************************************
I saw a fiber name recently that is unfamiliar to me and thought someone might help me out. It's "Marquesa Lana", no doubt a trade or brand name? Supposedly very durable as a chair fabric? Appreciate any leads.
} Hello: } Anybody who has the experience in using liquid helium stage for either } TEM or SEM, or has the information on where I can find this type of } stage, please send me an email at wtao-at-mtu.edu. Thanks. } } Weimin Tao
Oxford Instruments supply both SEM and TEM liquid He stages. Gatan Inc supplies a liquid He TEM stage.
When I worked for Gatan, I used their TEM He stage a few times. The biggest hassle was actually arranging the He supply and cooling the stage down. Once you'd done that, I didn't see any real problems using it, although I never spent a lot of time working with it.
Hello all; We have developed some problems with heat polymerization of our GMA for light microscopy, and would like to do some testing of UV polymerization. One of my co-workers remembers that in another lab she worked in, distance from the light source was an important factor. I believe Wayne England, who used to be at Carleton University, worked out a lot of bugs on this system... some input from him, and anyone else on the list would be most welcome. If you feel the information is not of interest to the general list, please contact us directly:
Lu-Ann Bowman bowmanla-at-em.agr.ca or Shea Miller millers-at-em.agr.ca
Thanks in advance shea
p.s. Thanks a bunch for my SOS about in situ hybridization using DIG- we got some great suggestions/tips, and will be posting a summary to the list next week.
Dr. S. Shea Miller Agriculture & Agri-Food Canada Eastern Cereal & Oilseed Research Centre Rm 2068, Bldg 20, CEF Ottawa, Ontario Canada K1A 0C6 Phone: (613)759-1760 Fax: (613)759-1701 e-mail: millers-at-em.agr.ca
I am trying to obtain some price estimates of using PIXE (proton-induced X-ray emission) microanalysis. What does it cost per hour to use such a microprobe? Is it a lot costlier than using conventional microprobes?
Thanks a lot for your help -
Karin E. Limburg Dept. of Systems Ecology University of Stockholm, Sweden
I just did a quick web search using Alta Vista and found:
Marquesa lana is indeed a wear-resistant chair fabric (olefin) but I couldn't find a manufacturer's name.
Marquesa lana is also the name of a horse scratched in the 8th race at Sam Houston Race Park on March 30th. Perhaps she was rubbed too much and wore out?
Harry Crossman
} ---------- } From: David_R_Stadden-at-armstrong.com[SMTP:David_R_Stadden-at-armstrong.com] } Sent: Friday, May 16, 1997 8:12AM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Fiber ID? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I really appreciate very much all your answers, ideas and suggestions about mechanical and diffusion pumps fluids. We have asked quotations to different companies for differents brands (all of them taken from from your messages) and finally we have decided, according to the prices and facilities + duties to be paid to be imported in Argentina, to buy Fisherbrand #19 and Santovac 5. Thanks to your time and typing effort we have gotten more money than what we had expected. Thanks very much. Greetings from the very south of America.
Silvia Montoro Centro Regional de Investigacion y Desarrollo Santa Fe - Argentina
Position for an electron microscopy technician is availible at Emory University neurology department (Atlanta, Georgia). Minimum qualification required includes B.S in a related field and 1 year experience in an anatomical research lab. The technician will be involved in studies on Fragile X syndrome and Huntingtons disease. Routine work includes morphological and immunocytochemical sample processing for light and electron microscopy, ultramicrotoming, examining sections on electron microscope, and digitizing results on computer. The starting date of this job will be on July 1. 97. Interested individuals please send CV to the following address:
Hong Yi Rm 6223 Woodruff Memorial Building Department of Neurology Emory University 1639 Pierce Dr. Atlanta, GA 30322
Hi Kaye; I have experienced this problem as well, and haven't figured out where it comes from. Initially I thought it was age, (old sections) but have also had problems with freshly sectioned material, (including freshly embedded/polymerized). Something that seems to help... I soak the slide with attached sections in a coplin jar with acetone for 10 minutes or so (I have gone up to 30 or 40 minutes), rinse with water, and then apply my stain while the sections are still wet. This may be a problem for some of the stuff you want to look at. Generally the cracks seem to disappear, or are at least minimized, and it no longer interferes with the microstructure of whatever I'm looking at. Hope this helps shea Dr. S. Shea Miller Agriculture & Agri-Food Canada Eastern Cereal & Oilseed Research Centre Rm 2068, Bldg 20, CEF Ottawa, Ontario Canada K1A 0C6 Phone: (613)759-1760 Fax: (613)759-1701 e-mail: millers-at-em.agr.ca
Hello! Does anyone have a used Ion Tech ion mill lying around that needs a good home? I have a personal project that requires one. Please reply directly - thanks!
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Page 6773 of the tenth edition of the Merck Index states the uses of osmium tetroxide as "an oxidizing agent " , "catalyzes chlorate, peroxide, periodate and other oxidations". Osmium is used "as an alloy with iridium for pen points and fine machine bearings; as catylyst in the synthesis of ammonia ; as catylyst in hydrogenation of organic compounds ". I suggest that could explain the presence of Os in SF bay from industrial uses. Another explanation is the incredible sensitivity of the detection equipment. How much is more that expected? Kate Connolly
Karin I have no experience of using the technique myself but I met a man who did. If I recall correctly the cost was higher because of the need of a proton source and the specialist design of the associated equipment. There was a group at Oxford 5 years ago (I assume they are still there) and you could try contacting: F. Watt, Scanning Proton Microprobe Unit, Nuclear Physics Laboratory, Department of Physics, Keble Road, Oxford OX1 3RH, UK. I'm sorry that I don't have a phone, fax or e-mail address but I'm sure that there will probably be a Web site. Malcolm Haswell University of Sunderland UK ----------
llo microscopists,
I am trying to obtain some price estimates of using PIXE (proton-induced X-ray emission) microanalysis. What does it cost per hour to use such a microprobe? Is it a lot costlier than using conventional microprobes?
Thanks a lot for your help -
Karin E. Limburg Dept. of Systems Ecology University of Stockholm, Sweden
From Microscopy-request-at-sparc5.microscopy.com Fri May 16 11:07:51 1997 Received: from srvr20.engin.umich.edu (root-at-srvr20.engin.umich.edu [141.212.2.26]) by srvr5.engin.umich.edu (8.8.5/8.8.5) with ESMTP id LAA05680; Fri, 16 May 1997 11:07:50 -0400 (EDT) Received: from judgmentday.rs.itd.umich.edu (0-at-judgmentday.rs.itd.umich.edu [141.211.83.37]) by srvr20.engin.umich.edu (8.8.5/8.8.5) with ESMTP id LAA17361; Fri, 16 May 1997 11:07:49 -0400 (EDT) Received: by judgmentday.rs.itd.umich.edu (8.8.5/2.2) with X.500 id LAA13249; Fri, 16 May 1997 11:07:49 -0400 (EDT) Received: from Sparc5.Microscopy.Com by judgmentday.rs.itd.umich.edu (8.8.5/2.2) with SMTP id LAA13110; Fri, 16 May 1997 11:07:29 -0400 (EDT) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id JAA04028 for dist-Microscopy; Fri, 16 May 1997 09:42:03 -0500 Received: from imsd-exchange.nrc.ca (imsd-exchange.nrc.ca [132.246.160.7]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id JAA04023 for {microscopy-at-MSA.microscopy.com} ; Fri, 16 May 1997 09:42:01 -0500 Received: by imsd-exchange.nrc.ca with SMTP (Microsoft Exchange Server Internet Mail Connector Version 4.0.994.63) id {01BC61E6.2205D070-at-imsd-exchange.nrc.ca} ; Fri, 16 May 1997 10:44:30 -0400 Message-ID: {c=CA%a=_%p=NRC%l=NRC/IMSD/001293E4-at-imsd-exchange.nrc.ca} From: "McCaffrey, John" {John.McCaffrey-at-nrc.ca} To: worldwide BB for microscopist {microscopy-at-sparc5.microscopy.com} Subject: Used Ion Tech ion mills Date: Fri, 16 May 1997 09:48:00 -0400 X-Mailer: Microsoft Exchange Server Internet Mail Connector Version 4.0.994.63 MIME-Version: 1.0 Content-Type: text/plain; charset="us-ascii" Content-Transfer-Encoding: 7bit Errors-to: Microscopy-request-at-sparc5.microscopy.com Status: RO
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com -----------------------------------------------------------------------.
Hello! Does anyone have a used Ion Tech ion mill lying around that needs a good home? I have a personal project that requires one. Please reply directly - thanks!
From Microscopy-request-at-sparc5.microscopy.com Fri May 16 11:07:51 1997 Received: from srvr20.engin.umich.edu (root-at-srvr20.engin.umich.edu [141.212.2.26]) by srvr5.engin.umich.edu (8.8.5/8.8.5) with ESMTP id LAA05680; Fri, 16 May 1997 11:07:50 -0400 (EDT) Received: from judgmentday.rs.itd.umich.edu (0-at-judgmentday.rs.itd.umich.edu [141.211.83.37]) by srvr20.engin.umich.edu (8.8.5/8.8.5) with ESMTP id LAA17361; Fri, 16 May 1997 11:07:49 -0400 (EDT) Received: by judgmentday.rs.itd.umich.edu (8.8.5/2.2) with X.500 id LAA13249; Fri, 16 May 1997 11:07:49 -0400 (EDT) Received: from Sparc5.Microscopy.Com by judgmentday.rs.itd.umich.edu (8.8.5/2.2) with SMTP id LAA13110; Fri, 16 May 1997 11:07:29 -0400 (EDT) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id JAA04028 for dist-Microscopy; Fri, 16 May 1997 09:42:03 -0500 Received: from imsd-exchange.nrc.ca (imsd-exchange.nrc.ca [132.246.160.7]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id JAA04023 for {microscopy-at-MSA.microscopy.com} ; Fri, 16 May 1997 09:42:01 -0500 Received: by imsd-exchange.nrc.ca with SMTP (Microsoft Exchange Server Internet Mail Connector Version 4.0.994.63) id {01BC61E6.2205D070-at-imsd-exchange.nrc.ca} ; Fri, 16 May 1997 10:44:30 -0400 Message-ID: {c=CA%a=_%p=NRC%l=NRC/IMSD/001293E4-at-imsd-exchange.nrc.ca} From: "McCaffrey, John" {John.McCaffrey-at-nrc.ca} To: worldwide BB for microscopist {microscopy-at-sparc5.microscopy.com} Subject: Used Ion Tech ion mills Date: Fri, 16 May 1997 09:48:00 -0400 X-Mailer: Microsoft Exchange Server Internet Mail Connector Version 4.0.994.63 MIME-Version: 1.0 Content-Type: text/plain; charset="us-ascii" Content-Transfer-Encoding: 7bit Errors-to: Microscopy-request-at-sparc5.microscopy.com Status: RO
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com -----------------------------------------------------------------------.
Hello! Does anyone have a used Ion Tech ion mill lying around that needs a good home? I have a personal project that requires one. Please reply directly - thanks!
I gave the wrong fax number in the earlier version of this message. Sorry.
A colleague is interested in purchasing a used, inexpensive light microscope. Nothing fancy: standard LM with 10x, 40x and 100x objectives and reasonable resolution. To be used for examination of biological and/or clinical materials (smears, sections, etc). Private individuals or vendors should contact:
Contact: Dr. Faiqa Hassan St. Louis, Mo. Phone: 314-522-0083 Fax: 314-522-1179
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 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 Microscopy ListServer -- Sponsor: The Microscopy Society of America
I saw a fiber name recently that is unfamiliar to me and thought someone might help me out. It's "Marquesa Lana", no doubt a trade or brand name? Supposedly very durable as a chair fabric? Appreciate any leads.
} I am trying to obtain some price estimates of using PIXE } (proton-induced X-ray emission) microanalysis. What does it cost per } hour to use such a microprobe? Is it a lot costlier than using } conventional microprobes? } Hi Karin:
Try to contact the PIXE group (Prof. Malmqvist et al.) in Lund. You might get a free access. They are helpful. The address is: Nuclear Physics Fax: +46 46 222 4709 E-mail: nuclear-at-nuclear.lu.se
Regards,
B.E.
Benyam Estifanos Dept. of Mineralogy & Petrology University of Lund Soelvegatan 13 S-223 62 Lund, Sweden INTERNET: estif-at-gemini.ldc.lu.se / Benyam.Estifanos-at-Geol.lu.se Tel.:+46 46 2224597; Fax:+46 46 121477
YUHUI XU requested Info on DNA imaging; below is a few articles Ive found helpful....
SALLY BURNS e-mail: burnssal-at-pilot.msu.edu
REVIEW ARTICLES
Davis, R., Simon, M., and Davidson N. 1971 Electon Microscope Heteroduplex methods for Mapping regions of Base sequence homolgy in Nucliec acids Methods in Enzymology #21 419-428 (QP601.C733)
Burkhardt and Lurz Electron microscopy: CHP 6 in the book: Adv Molec Genetics 1984 (QH 430.A38 1984)
Spiess, E., and Lurz R. 1988 EM analysis of nucleic acids and nucleic acid protein complexes. Methods in microbiology 20: 293-323. ........................................................ ------------------------Other relevant articles
Backert, S., Lurz,R., and Borner T. 1996 E.M investigation of mitochondrail DNA from Chenopodium album (L.) Curr Genet 29:427-436
Backert, S. Dorfel, P., Lurz, R., and Borner, T. 1996 Rolling circle...... Mol and Cellular Biol. vol 16 no 11 p6285-6294.
DAVIS, RW AND HYMAN,RW. 1971 A study in evolution : ..... J. MOL BIOL 62,287-301.
Kolodner, RD and Tewari,K. 1974 Denaturation Mapping Studies on the Circular Chloroplast deoxyribonucleic Acid from Pea Leaves J Biological Chem 250313 4888-4895
Kolodner, RD and Tewari, K. 1975. Presence of Displacement loops..... J. Biol Chm vol 250, No22 8840-8847.
****LANG,D., and MITANI, M. 1970 ******** Simplified Quantitiative Electon Microsopy of Biopolymers Biopolymers Vol 9 373-379.
Wessel Muller, H. and Hoffman Berling (1990) ELECTRON MICROSOPY OF DNA-HELICASE I COMPLEXES IN THE ACT OF STRAND SEPERATION Eur J Biochm 189,277-285 ......... (uses SSB and formamide.)
I have just become the principal operator of a Zeiss DSM-960, and would like to hear any comments or insights from anyone having experience with Zeiss SEM's, especially pertaining to the use of LaB6 sources. I have experience on other vendors instruments, but the design of cathode assembly is different than what I am familiar with. Specifically, there is no height adjustment, other than a shim. The instrument manuals provide little information and simply state the LaB6 is not operator replacable and is factory serviced as an assembly. The lab has been sending the entire assembly to FEI for rebuilding. Is this really necessary? Does anyone replace their own sources on a Zeiss?. I can only find reference to a Zeiss base for the DENKA M1 and M7 in any of the supplier catalogs I have, although I haven't talked to any suppliers yet.
Thanks Jim Mabon _____________________________________________________ James C. Mabon Center for Microanalysis of Materials Frederick Seitz Materials Research Laboratory 104 South Goodwin Avenue Urbana, Illinois 61801 (217)333-4265 *Fax(217)244-2278 email: mabon-at-mrl7.mrl.uiuc.edu {that's the letter l} _____________________________________________________
With regard to the question from Cam on the matter of Be grids: ------------------------------------------------------ I am concerned, really, about the toxicity of that material. Are there any viable alternatives? Or is the hazard, to operator or environment, in fact not so worrisome that anyone really need be concerned about looking for something else? (Opinions?) I'm also interested to learn for what work it is generally felt that one should even be considering a beryllium grid, since they are pretty costly to boot! ------------------------------------------------------
and the reply from Larry Stoter: ------------------------------------- There are alternatives - Al grids resolve most of the above problems. Carbon coated nylon and carbon composite grids are also available from the usual suppliers - Agar, SPI, etc. However, for really critical analysis, I'd say Be grids are to be preferred.
I don't believe there is any serious hazard associated with these grids. Be is a problem if it get into the lungs (or trapped under the skin), where it leads to medical conditions similar to those caused by asbestos. So don't use abrasives on it, and I guess you want to avoid exposure to any strong acids that might attack the oxide surface (and don't eat them:) =================================================
There are "alternatives", but most really are not as acceptable as Be. The Al grids give a strong x-ray line that can swamp the system much like Cu and also, coatings (e.g. "Formvar", etc.) don't seem to want to stick as well as on Be. Oxide coatings on the Al tend to reduce conductivity even further. The carbon composite grid, while conductive, gives high Bremsstrahlung background radiation reducing experimental sensitivites. The Nylon grids have to be carbon coated to reduce charging, but generally it is never eliminated completely and charging does continue to be at least somewhat of a problem. Also, the Nylon grids (at least the ones we have tried) are further plagued by a low level additive of TiO2 which gives additionally, low level instensities due to Ti. And there is still the problem with the high Bremsstrahlung background radiation. Probably diamond grids come the closest to being an acceptable substitute, and the Bremsstrahlung background is not significant, except that they are even more expensive than Be. But for those who operate in institutional environments where Be is not permitted under any circumstances, the diamond grids would be the alternative of choice, at least technically, even if it would fail economically.
Now a few other comments about Be grids:
a) what is toxic is the oxide, not the metal. There is widespread appreciation that BeO is highly toxic. However, what is not toxic is the metal itself. So if what is being done, in the handling of the Be grids does not result in the formation of an oxide scale or film, something that could in theory, "come off", then so far as we know, there is no problem. I am unaware or any explanation of how the oxide could form while being irradiated in the column and I am unaware of anyone who has ever shown that the oxide has indeed formed on any of the Be grids.
I myself feel much more comfortable offering for sale the Be grids, for example, than our Be planchettes. While it is true that (at least our) planchettes are accompanied with all kinds of safety instructions, the fact is, I have this nagging concern that a Be planchette once carbon coated, for example, tends to take on the appearance like any other planchette or mount made of aluminum, and very clearly, as evidenced by the recent postings on the topic of the cleaning of SEM mounts, there is quite a bit of grinding and recycling of Al mounts. Hence if one or more Be planchettes ended up accidentally being mixed in with the Al mounts, and received these same "grinding and polishing" treatments, then there very well could end up being a BeO dust problem, from the grinder and polisher for sure when they became dried out.
Now, from my own personal contacts in the Be industry (remember, such contacts might not give out objective information!), I have been told that even in the case of the drying out of a polishing table, the levels of dust released would be at such low levels that it would be an extraordinary thing for someone to suffer any of the classic symptoms of berylliosis. However, they do point out that some small percentage of us are in fact highly reactive to such low levels, and the problem is that no one knows in advance who is going to be a reactor and who would not react.
Quite possibly, at least some of the concern about the grids flows from this very real concern related specifically to the planchettes but which would not be applicable to the grids since grids are not "ground and polished".
b) With regard to the "high cost", there is nothing that can be done about that, however, from all reports we have ever heard, as well as our own experience in our own laboratory, Be grids in a small beaker of acetone in an ultrasonic shaker for some period of time, tends to result in a fairly quick cleaning up for reuse.
Also, plasma cleaning as a final step, is a possibility as well, however, in this instance I am a bit less certain. Some years ago, we did an oxygen plasma exposure of several minutes to some Be grids and found no change. This was not a rigorous "looking" for oxide formation, but to the eye, and also, at stereo zoom LM level there was no apparent change. In other laboratories this seems to be a standard way of cleaning for the reuse of Be grids. So while there might be "sticker shock" when Be grids are first purchased, the ability to reuse them over and over again, tends to mitigate that high cost and furthermore, the enhancement to data quality (as opposed to the alternatives) is quite significant to say the least.
c) All Be grids were not created equal, so the results that are obtained from one supplier may or may not be the same as from all other suppliers. I am referring to both the quality of the etching of the Be foil and also, the purity of the Be starting foil material. So far as I know, the Be grids from Agar, as well as those from SPI, are made from the highest purity Be foil starting material available for this kind of use.
In any case, we are frequently asked about these alternatives and there is no one place, at least that I am aware of, where one could find the "answers".
Disclaimer: SPI Supplies is a supplier of Be grids, diamond grids, as well as the other grids mentioned so it would be in our own best interests to see more people doing this kind of work.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Can anyone refer a source for laser printer enhancement software, similar to LaserPix, which is compatible with WINDOWS NT 32 bit architecture? Now using HP Laserjet 4.
Thanks for the help,
Bob Hessler Hessler Technical Services Stamford, CT PHONE/FAX: (203) 358-0266 E Mail: hestec-at-ix.netcom.com
Sorry to bring up this subject again but we have just managed to get the = funds to purchase a negative scanner. We need to scan 35mm, 120 and TEM = (6.5x9cm) negatives so we need a versatile system which will run on a = PC.
From what I have been able to get so far, dedicated negative scanners = are better than flatbeds with transparency adaptors, so we've decided to = restrict ourselves to these. However if you know of any flatbeds which = match the performance of the dedicated ones please respond.
There are 3 which I have managed to find available in Australia within = our price range. These are:
Nikon LS 4500 AF - had some problems but I've been assured that these = have been fixed
Kodak RFS 3570 - don't know much about these
Polaroid SprintScan 45 - don't know much about these either.
So, if you have had any experience (good or bad) running any of the = above on a PC, or if you know of any other suitable scanners please tell = me.
Many thanks in advance.
Colin Veitch
Colin.Veitch-at-dwt.csiro.au CSIRO Division of Wool Technology PO Box 21 Belmont Vic 3216 Australia Tel: +61 (0) 3 5246 4000 Fax: +61 (0) 3 5246 4057
At 08:42 AM 5/15/97 -0600, Tobias Baskin asked: } I wonder anyone knows of a source for Technovit resin in the USA? } This resin is made by Hareus (spelling?) in Europe. Is there perhaps a } similar if not identical resin available here? Also, does anyone know if } Technovit is glycol methacrylate?
We (Energy Beam Sciences) took over distribution of the Technovit resins from Delaware Diamond Knives about two months ago. They are manufactured by Heraeus Kulzer in Germany. There are three resins:
Kulzer has given us some very nice technical/applications brochures on these products; please contact me back-channel if you would like one.
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
Dear colleagues, I would like to add two works to the "DNA biblio" list:
Preparation and length measurements of the total deoxyribonucleic acid content of T2 bacteriophages. 1962 (classical article) Kleinschmidt-AK; Lang-D; Jacherts-D; Zahn-RK Biochim-Biophys-Acta 1989; 1000: 41-48
Preparation and length measurements of the total DNA content of T2 bacteriophages: a commentary on 'Darstellung und Langenmessungen des gesamten Desoxyribonucleinsaure-Inhaltes von T2-Bakteriophagen' [published erratum appears in Biochim Biophys Acta 1989 Dec 8; 993 (2-3):309] Kleinschmidt-AK Biochim-Biophys-Acta. 1989; 1000: 35-40
A lot about beginning of EM of DNA can be found in these articles.
Best regards O. Benada +---------------------------------------------------------------+ Dr. Oldrich Benada Acad. Sci. CR Phone: +420-2-4752399 Institute of Microbiology Fax: +420-2-4715743 Videnska 1083 E-mail: benada-at-biomed.cas.cz 142 20 Prague 4 - Krc Czech Republic +---------------------------------------------------------------+
Dear colleagues, I would like to add two articles to the "DNA biblio" list:
Preparation and length measurements of the total deoxyribonucleic acid content of T2 bacteriophages. 1962 (classical article) Kleinschmidt-AK; Lang-D; Jacherts-D; Zahn-RK Biochim-Biophys-Acta 1989; 1000: 41-48
Preparation and length measurements of the total DNA content of T2 bacteriophages: a commentary on 'Darstellung und Langenmessungen des gesamten Desoxyribonucleinsaure-Inhaltes von T2-Bakteriophagen' [published erratum appears in Biochim Biophys Acta 1989 Dec 8; 993 (2-3):309] Kleinschmidt-AK Biochim-Biophys-Acta. 1989; 1000: 35-40
A lot about beginning of EM of DNA can be found in these articles.
Best regards O. Benada +---------------------------------------------------------------+ Dr. Oldrich Benada Acad. Sci. CR Phone: +420-2-4752399 Institute of Microbiology Fax: +420-2-4715743 Videnska 1083 E-mail: benada-at-biomed.cas.cz 142 20 Prague 4 - Krc Czech Republic +---------------------------------------------------------------+
If anyone can help Matt out, please reply to him directly
} From: Abrcrombee-at-aol.com } Date: Sat, 17 May 1997 21:25:53 -0400 (EDT) } To: sbarlow-at-sunstroke.sdsu.edu } Subject: Hello } } I found your email address while searching for information on Trichonympa. } My biology teacher assigned my class semester reports, and mine is on that. } Would you possibly have any information that you could send me about } Trichonympa? I would appreciate it if you would send something.....anything } at all, for I have found barely anything on this organism. Thanks...Matt } McCluskey }
Dr. Steven Barlow EM Facility/Biology Department 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/EM_Facility
I do not purport to be an expert on Be but we do handle a fair amount of it around here for other purposes than TEM grids. I believe your response may promote too cavalier an attitude towards handling Be as opposed to BeO. Our ES&H people do not make a distinction between the two regarding handling and toxicity. The latest MSDSs I have for the two indicate roughly the same hazards (NFPA ratings of 2,0,0 for BeO and 2,1,1 for Be). The toxicological sections of the MSDS also indicate roughly the same hazard, around 20mg/kg producing tumors in rats. While I agree that on the scale of all hazardous things in the world, Be is only a moderate hazard for most people (the ~95% who are not allergic), I would be more cautious as regards recycling Be grids. I think they {italic} should be {/italic} recycled to reduce the volume of Be waste generated, but people should pay particular attention to any operation that has the potential for producing respirable dust. Our people who produce sputtered Be films must wear full face respirators every time they open the vacuum chamber even though the material is deposited as a film on the inside of the chamber and not very likely to produce dust.=20 As an aside we have noted that sputtered Be films are fairly reactive, almost always producing substiochiometric oxide, which may not necessarily be applicable to the TEM grid case, but is some indication of its reactivity.
We have a Reichart Ultracut E with the cryo attachment FC 4. The flange on the liquid nitrogen vessel which is used to attach the corresponding flange on the refill dewar head has developed a (nitrogen) leak. The leak developed in the joint between the flange and the dewar . We actually located the point where it leaks . We tried adding some epoxy to that area, but it did not solve the problem.This joint seems to be an epoxy . The manufacturer wants to replace the whole dewar at considerable cost. Has anyone encountered a similar problem . I would appreciate any comments and possible (alternate) solutions.
In a number of German microtechnique texts, including D. Gerlach's "Botanische Mikrotechnik", I have seen references to Pfeiffer's Mixture as being ideally suited to fixing of algae, particularly the filamentous forms. However, I'm confused about one of the constituents of this mixture, given in German as "roher Holzessig". I thought this might be the same as tannic acid, but I'm not sure. I'm also not sure of the concentration and the meaning of the term "roher" (raw) in the description.
Can anyone help clarify the formula Pfeiffer's Mixture? I have not found any references to Pfeiffer's Mixture in any English or American texts on botanical microtechnique - are there fixing mixtures which are as good or better for filamentous algae?
I have several thousand confocal images (TIFF single files, 8bit grey, 24 bit RGB, z-series etc.) and I was wondering whether there is some relatively cheap software/shareware around that I could use to catalogue the images. I want to be able to save some information with the images and thumnail sketches would be useful.
Alternatively, if someone knows how I can use Microsoft Access as an image database without having to embed the full resolution picture in each record I would be grateful.
Any ideas? contact Mark Auty Mark Auty DPC Moorepark Fermoy Co. Cork Ireland
As this is perhaps the very first enquiry on PIXE microanalysis, let me try to give a few details.
Firstly, you were given the mail address to Lund nuclear microprobe group. The full address is: Dept. of Nuclear Physics, Lund Institute of Technology, Box 118, SE-221 00 Lund, Sweden In addition to the e-mail which was given to you, the contact with Jan Pallon (one of the senior members of this group) is: Pixejan-at-outis.lucas.lu.se or: Jan.Pallon-at-pixe.lth.se
To answer directly your question on running costs. At our nuclear microprobe, which is situated in South Africa, at National Accelerator Centre in Faure, close to Cape Town, the full cost is ca. 100 USD per hour. This is however the maximum price, for industrial applications with full confidentiality of results, if required. We are the so-called national facility and users from local universities are encouraged to apply for the beam time, and in many cases this comes free of charge. Our preferred collaboration is under projects in many research fields (geology, materials science, botany and life sciences). This means longer collaboration, sharing students if possible, full access to data. We welcome any international collaboration, if suitable for this type of equipment.
You can contact me at:
PRZYBYLOWICZ-at-nac.ac.za Address: Dr. W.J. Przybylowicz National Accelerator Centre Van de Graaff Group P O Box 72 Faure 7131 South Africa Fax: +27-21-8433543
Web site (does not give all details; some information, e.g. publications, is not fully updated) is: http://www.nac.ac.za xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx Dr Wojciech J. Przybylowicz 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 xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx
I run a morphology core where a TEM is used every day. Last year when I was pregnant, I called Health Physics who monitored the scope and me for a period of time. There was absolutely no leakage fromm the column. Although every scope is different, ours is brand new, you can always have your environmental people come out and check for leaks. I believe them to be safe - my son had all of his fingers and toes.
Cheri Owen
On Tue, 13 May 1997, PATRICK DIEHL wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Do any of the universities, companies, hospitals or research institutions } have any policies regarding the use of electron microscopy by pregant } employees? Is there any literature available on the topic? } } Is the advice not to use EM at all during the 9 months, or maybe only } during critical periods of development? } } In searching the archives I only found one reference on this topic. } } Any help would appreciated. } } Patrick Diehl } Materials Science Department } University of North Texas }
If you have a single or double tilt heating holder (for Philips 400 series microscopes)that is in working order and you want to part with it, contact me privately so we can discuss arrangements.
John C. Wheatley Lab Manager Center for High Resolution Electron Microscopy BOX 871704 Tempe, AZ 85287-1704 Phone: (602) 965-3831 FAX: (602) 965-9004 John.Wheatley-at-ASU.Edu
Thank you to all who have responded to my e-mail. If anyone wants copies of the replies, please e-mail me privately. I am unsubribing from the microscopy list.
I am anxiously seeking microscopy video footage for a project which deal with
blood pressure, the contraction of blood vessels and arteries and catherters imagery. Specifically, I need: -blood traveling through arteries (not capillaries) -footage from inside arteries -video from catharter: somethind like a 3/4" view which illustrates pressure which closes and opens valves (looks like open and closing circle) - perhaps this is from an endoscopy?
Please contact me at mhallmedia-at-aol.com with any information you may forward. Many thanks, Heather
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I'm in need of some advice on delineating grain boundaries in a plated copper film. I've tried several ASTM wet etches and a few different CF4 plasma etches, but am not satisfied with the results. Grain boundaries are somewhat defined but not clearly enough to confidently measure their parameters. Does anyone have an etch they are satisfied with for this application? I'm using an FESEM for imaging and have had very good results delineating various alloys, but this copper film of roughly 7 microns thick has me puzzled. X-ray diffraction shows the film is crystalline. Any help is greatly appreciated.
********************************************************** Jake Schaper Product Analysis Lab Advanced Digital Consumer Division Motorola, Inc. 1300 N. Alma School Rd. Chandler, Arizona 85224 Mail Drop CH240 Phone 602-814-4756 **********************************************************
Hi: Our school have received in donation recently a sphectrophotometer BECKMAN 24, that has arrived whitout the technicals handbooks. If some people can send us a copy of all , please contact me thanks in advance =================================================== Fernando D. Balducci Laboratorio de Microscopia Electronica Facultad de Ingenieria - Bioingenieria Universidad Nacional de Entre Rios Argentina e-mail: microsc-at-fi.uner.edu.ar tel: 54 43 975100 fax: 54 43 975077 ===================================================
Our department has 2 academic positions currently advertised of which it is intended that one will be filled by a person with an active interest in cell biology and in particular the development of microscopy related research techniques in cell biology (ie Confocal Microscopy, Electron Microscopy, Image Processing and Anaylsis). A copy of the advertisement follows;
LECTURERS IN ANATOMY AND STRUCTURAL BIOLOGY
Two lectureships in the Department of Anatomy and Structural Biology will be available from September 1997. These are confirmable positions. Appropriately qualified applicants could be considered for appointment at Senior Lecturer level.
Applicants will be expected to hold a PhD or equivalent qualification, be committed to research, have proven research ability, and have an on-going and active research program. Their interests should fit within one of the areas of research excellence within the Department: neuroscience, cell biology, reproductive biology, developmental biology and functional anatomy.
The successful applicants will also play an active role in teaching. The Department has teaching commitments to medical, dental, physiotherapy, medical laboratory science, pharmacy, physical education and science students. The appointees will teach in two or more of the following areas: cell biology, histology, gross/functional anatomy, reproductive biology, and/or neuroscience, at both the undergraduate and postgraduate level.
The Department will be looking to make one appointment in the area of cell biology and one in functional anatomy (with responsibility for physiotherapy teaching).
Further information about the position and the Department can be obtained from Professsor DG Jones, Department of Anatomy and Structural Biology (Tel: 64 3 479 7364; Fax: 64 3 479 7254; email: gareth.jones-at-stonebow.otago.ac.nz) and/or from the Department's homepage: http://gandalf.otago.ac.nz:800/Anatomy/home-page.html.
Salary of Lecturer (Non-medical): $NZ42,750 - $53,250 Salary of Senior Lecturer (Non-medical): $NZ56,250 - $66,250
Reference number A97/32. Closing Date 4 July 1997.
METHOD OF APPLICATION
Further details regarding this position, the University, and the application procedure, are available from the Deputy Director, Personnel Services, University of Otago, PO Box 56, Dunedin, New Zealand (facsimile 64-3-474 1607 or e-mail laurie.hibbert-at-stonebow.otago.ac.nz).
Applicants should send two copies of their curriculum vitae together with the names, addresses and fax numbers of three referees, to the Deputy Director of Personnel Services by the specified closing date, quoting the appropriate reference number.
If an applicant is short listed for interview, whanau support will be welcome.
Equal opportunity in employment is University policy.
E tautoko ana Te Whare Wananga o Otago i te kaupapa whakaorite whiwhinga mahi.
One of our students is doing a project involving the production of artificial caries on teeth, in vitro. She would like to use Quinoline as one of the stains to help delineate the structure of the lesion but we cannot find any information on what concentration to use. Articles which mention Quinoline do not give any information as to what concentration was used.
To all UK TEM users We have a Philips 301 TEM here at the dept. of Materials Engineering & Materials Design, Nottingham University, that is surplus to our requirements. This instrument has been out of use for several years and we need the space it is taking up for a new ESEM-FEG. The TEM will need some repairs to get it operational again and may be most suitable for spare parts. We urgently need to move this microscope out of the department and are prepared to give it away, if you are prepared to pay removal costs.
All interested parties please contact Ms.Nicola Bock, Dept.of ME&MD,University of Nottingham. Tel: (0115)9513759 Email: emznjb-at-emn1.nott.ac.uk
I use PC-Image Release Beta 1 under Windows NT 4.0. When I change the clipboard size from the default setting 400k in Options/Preferences NIH-image terminates. What might be the reason? Any ideas?
We are interested in purchasing a portable multi-media projector to present microstructures, either in form of digital images or in form of .avi (or similar) movie clips from in-situ SEM or TEM analysis. Is the expense to go from Polysilicon to DLP technology worthwhile? How portable are they really, i.e. how easy it is to set up the display at any location? How much of the details of an image is lost, if any?
Thanks for the help Hasso Weiland Alcoa Technical Center Alcoa Center, PA 15069
Hi guys, Does anyone know of any shareware that will allow me to convert quicktime movies into individual frames. Thanks in advance,
--Ciprian Have fun and keep the sun on your back and a smile on your face. __________________________________________________________ Ciprian A. Almonte University of Pittsburgh Center for Biologic Imaging Pittsburgh, PA 15261
Visit my web site at http://www.pitt.edu/~calmonte Laboratory's website: http://sbic6.sbic.pitt.edu __________________________________________________________
You could try GifBuilder - this is used for making animated GIF89 image files, but it can read in QuickTime movies and export single frames as GIF files (which can then be converted to other formats with Photoshope, etc...). You can find info and download the program at
http://member.aol.com/royalef/toolbox.htm
This has links to other software as well. Hope it's useful,
Rick Powell
****************************************************************** * NANOPROBES, Incorporated | Tel: (516) 444-8815 * * 25 East Loop Road, Suite 124 | Fax: (516) 444-8816 * * Stony Brook, NY 11790-3350, USA | nano-at-mail.lihti.org * * * * NOW EASY TO FIND ON THE WEB: http://www.nanoprobes.com * ******************************************************************
Our department has 2 academic positions currently advertised of which it is intended that one will be filled by a person with an active interest in cell biology and in particular the development of microscopy related research techniques in cell biology (ie Confocal Microscopy, Electron Microscopy, Image Processing and Anaylsis). A copy of the advertisement was posted yesterday;
LECTURERS IN ANATOMY AND STRUCTURAL BIOLOGY UNIVERSITY OF OTAGO DUNEDIN NEW ZEALAND
Sorry, the institution at which these positions were available was not posted yesterday.
I need help fighting the bean counters! We are currently working our way thru a new cost recovery evaluation. Based on the results off this evaluation I am being asked to raise my rates for Basic TEM and SEM services to a level which seems outrageously high to me. I would like to find out what other labs are charging for scope time, as well as basic sample preperation. If you don't want to post this information to the listserv you could E-mail me directly at fnksd1-at-aurora.alaska.edu Thank you! Kim
I just borrowed an Apollo system for a local microscopy meeting. (Sorry, I don't know the model number or where they bought it.) We used it to interactivelly display a Photoshop 4.0 demonstration (1024x780). It has a remote mouse joystick with a laser pointer in it. The mouse operated from about 25-30ft by pointing it at the screen and bouncing it into the unit. We operated it off a laptop. We also used it to play output from a VCR unit onto the screen with sound. (BTW, I highly recommend MSA's LAS speaker, Conly Rieder to local societies-He comes with a great understandable video presentation with sound.) The unit weighed about 25 pounds and was easily transported by its handle. ( I carried it a long way across a parking lot and also carried it on a bus without any trouble or pausing for breath.
This thing was just fantastic. I understand that the price tag is about $13,000 though.
Let me know if you need details, I'll try to get them from our computer guys.
- -Scott Walck } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I just borrowed an Apollo system for a local microscopy meeting. (Sorry, I don't know the model number or where they bought it.) We used it to interactivelly display a Photoshop 4.0 demonstration (1024x780). It has a remote mouse joystick with a laser pointer in it. The mouse operated from about 25-30ft by pointing it at the screen and bouncing it into the unit. We operated it off a laptop. We also used it to play output from a VCR unit onto the screen with sound. (BTW, I highly recommend MSA's LAS speaker, Conly Rieder to local societies-He comes with a great understandable video presentation with sound.) The unit weighed about 25 pounds and was easily transported by its handle. ( I carried it a long way across a parking lot and also carried it on a bus without any trouble or pausing for breath.
This thing was just fantastic. I understand that the price tag is about $13,000 though.
Let me know if you need details, I'll try to get them from our computer guys.
- -Scott Walck } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I just borrowed an Apollo system for a local microscopy meeting. (Sorry, I don't know the model number or where they bought it.) We used it to interactivelly display a Photoshop 4.0 demonstration (1024x780). It has a remote mouse joystick with a laser pointer in it. The mouse operated from about 25-30ft by pointing it at the screen and bouncing it into the unit. We operated it off a laptop. We also used it to play output from a VCR unit onto the screen with sound. (BTW, I highly recommend MSA's LAS speaker, Conly Rieder to local societies-He comes with a great understandable video presentation with sound.) The unit weighed about 25 pounds and was easily transported by its handle. ( I carried it a long way across a parking lot and also carried it on a bus without any trouble or pausing for breath.
This thing was just fantastic. I understand that the price tag is about $13,000 though.
Let me know if you need details, I'll try to get them from our computer guys.
- -Scott Walck } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I just borrowed an Apollo system for a local microscopy meeting. (Sorry, I don't know the model number or where they bought it.) We used it to interactivelly display a Photoshop 4.0 demonstration (1024x780). It has a remote mouse joystick with a laser pointer in it. The mouse operated from about 25-30ft by pointing it at the screen and bouncing it into the unit. We operated it off a laptop. We also used it to play output from a VCR unit onto the screen with sound. (BTW, I highly recommend MSA's LAS speaker, Conly Rieder to local societies-He comes with a great understandable video presentation with sound.) The unit weighed about 25 pounds and was easily transported by its handle. ( I carried it a long way across a parking lot and also carried it on a bus without any trouble or pausing for breath.
This thing was just fantastic. I understand that the price tag is about $13,000 though.
Let me know if you need details, I'll try to get them from our computer guys.
- -Scott Walck } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I just borrowed an Apollo system for a local microscopy meeting. (Sorry, I don't know the model number or where they bought it.) We used it to interactivelly display a Photoshop 4.0 demonstration (1024x780). It has a remote mouse joystick with a laser pointer in it. The mouse operated from about 25-30ft by pointing it at the screen and bouncing it into the unit. We operated it off a laptop. We also used it to play output from a VCR unit onto the screen with sound. (BTW, I highly recommend MSA's LAS speaker, Conly Rieder to local societies-He comes with a great understandable video presentation with sound.) The unit weighed about 25 pounds and was easily transported by its handle. ( I carried it a long way across a parking lot and also carried it on a bus without any trouble or pausing for breath.
This thing was just fantastic. I understand that the price tag is about $13,000 though.
Let me know if you need details, I'll try to get them from our computer guys.
- -Scott Walck } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I just borrowed an Apollo system for a local microscopy meeting. (Sorry, I don't know the model number or where they bought it.) We used it to interactivelly display a Photoshop 4.0 demonstration (1024x780). It has a remote mouse joystick with a laser pointer in it. The mouse operated from about 25-30ft by pointing it at the screen and bouncing it into the unit. We operated it off a laptop. We also used it to play output from a VCR unit onto the screen with sound. (BTW, I highly recommend MSA's LAS speaker, Conly Rieder to local societies-He comes with a great understandable video presentation with sound.) The unit weighed about 25 pounds and was easily transported by its handle. ( I carried it a long way across a parking lot and also carried it on a bus without any trouble or pausing for breath.
This thing was just fantastic. I understand that the price tag is about $13,000 though.
Let me know if you need details, I'll try to get them from our computer guys.
- -Scott Walck } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I hope, that someone on the list can help me to find suitable PC-software (DOS or Windows) or other tools for the following problem: I have to reconstruct the three-dimensional structure of different organs of microarthropods based on serial sections of resin embedded specimen. Up to now I go the time consuming and not exact way of drawing complete transverse, saggital and horizontal series and then to "reconstruct" the 3D-structure more or less "by hand". Any idea is welcome. Heike Dr. Heike Buecking University of Bremen UFT Physiological Plant Anatomy Leobener Str. D 28359 Bremen Germany TEL: +49-421-218-2954 or TEL: +49-421-218-7283 FAX: +49-421-218-3737 e-mail: heibueck-at-uft.uni-bremen.de
} } I use PC-Image Release Beta 1 under Windows NT 4.0. } When I change the clipboard size from the default setting 400k in } Options/Preferences NIH-image terminates. } What might be the reason? Any ideas? } } Reinhard } } I had the same problem with the WIN95 version. I overcame it by manually editing the .ini file where buffer size= XXXX Hope this helps
Chris
Chris Gilpin Biological Sciences Electron Microscope Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 161 275 5170 fax +44 161 275 5171
Polysilicon offers a couple of benefits. The colors are truer, and the units are brighter (higher lumens/$). The DLP technology offers smoother images(colors/tones and edges) without the 'screen door effect' that is inherent with LCD technology(not a defect, just a function of the technology that enables you to see each and every pixel on the screen). This smoothness is more apparent in video with movement than a digital signal.
The units are portable in that they have handles and/or cases(generally with wheels) available. As far as setup goes, they are plug and play, just as if you are plugging a monitor. It is necessary to sync them with a digital signal since computers have different signals. This is most easily done with a checkerboard(black and white) wallpaper pattern.
The image that is projected is the same image/signal that you would see on your monitor. In some cases, depending on the specs of your monitor, it may actually be better. Most projectors have controls to adjust the picture, similar to monitors.
Another opportunity to consider is resolution. The demand for VGA(640 x 480) has fallen off and the current demand is highest for SVGA(800 x 600). There is some demand for XGA(1020 x 768) and SXGA(1280x 1040). Depending on the output of your computers currently and for the foreseeable future, you need to look at this as well. Generally, projectors are downwards compatible in resolution. There is a new technology, 'intelligent' or 'smart' compression that offers display of XGA signals on a unit that is technically SVGA. To keep it simple, this is essentially 'loss-less' compression of the XGA signal. I am aware of it only being available on DLP models.
As I suggest with any imaging system or device, have a dealer(s) bring in the two technologies and see for yourself. Specifications are great to compare but 'a picture is worth...'
John D. Warren Area Sales Manager Digital Products Polaroid Corporation "See What Develops" 4525 Leonard Parkway Richmond, Virginia 23221-1809 804 254 1011 804 254 1013 Fax warrenj1-at-polaroid.com
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We are interested in purchasing a portable multi-media projector to present microstructures, either in form of digital images or in form of .avi (or similar) movie clips from in-situ SEM or TEM analysis. Is the expense to go from Polysilicon to DLP technology worthwhile? How portable are they really, i.e. how easy it is to set up the display at any location? How much of the details of an image is lost, if any?
Thanks for the help Hasso Weiland Alcoa Technical Center Alcoa Center, PA 15069
Try checking Alcheny Mindworks (Yahoo will turn up the address). If I Recall correctly, they offer something along that line as inexpensive shareware..
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi guys, Does anyone know of any shareware that will allow me to convert quicktime movies into individual frames. Thanks in advance,
--Ciprian Have fun and keep the sun on your back and a smile on your face. __________________________________________________________ Ciprian A. Almonte University of Pittsburgh Center for Biologic Imaging Pittsburgh, PA 15261
Visit my web site at http://www.pitt.edu/~calmonte Laboratory's website: http://sbic6.sbic.pitt.edu __________________________________________________________
AutoQuant Imaging Inc. is a world leader in the research and development of deconvolution software products for 3D visualization in biology and ophthalmology. AutoQuant is a growth company with excellent opportunities for future advancement, and is planning a major expansion. We have the following 2 openings at our Troy NY laboratory.
Description: The candidate is expected to have completed his/her graduate degree in Electrical Engineering, Biomedical Engineering, or a closely related field from an accredited university. A background is preferred in biomedical imaging, although strong candidates with other qualifications listed below will be considered as well. All levels of experience will be considered. Responsibilities will be a diversity of three areas having equal emphasis: (1) Basic research, (2) product development, and (3) customer support. The candidate must have strong writing skills and strong interpersonal skills. Industrial experience will be considered a plus. A technical background or desire to work in any of the following areas will be considered a plus: graphical software development using C/C++, MFC and Visual C++, signal processing, image processing, mathematical modeling, optimization methods, detection-and-estimation methods. Although not essential, a background in optics, 3D microscopy, confocal microscopy, ophthalmic imaging, or radiological imaging (MR, PET, CT) will be considered a plus as well. The ideal candidate will be a self-starter, self-motivated, highly responsible, organized, capable of being self-managed and will require very little supervision. He/she will be diverse and capable of handling a multitude of projects, tasks and responsibilities at once.
The 2 openings are as follows: (1) R&D Scientist: An individual having a Ph.D. or equivalent degree and matching the above description. This individual will be responsible for leading new research initiatives, creating new ideas and for applying for external R&D contracts. A strong publication record or record of presentation at professional conferences is expected. (2) R&D Engineer: A creative and resourceful individual having a Masters degree and matching the above description. A strong publication record or industrial experience will be a plus, although not absolutely required.
AutoQuant is located adjacent to the Rensselaer Polytechnic Institute campus and collaborates in research with the Rensselaer Polytechnic Institute, the Wadsworth Center for Laboratories and Research and the Albany Medical College. The R&D Scientist will be expected to develop similar collaborations with these or other collaborating institutions. The opportunity exists for the R&D Engineer to pursue a Ph.D. degree on a part time basis.
The City of Troy is part of the historic tri-city Capital District of Upstate New York. The area features numerous recreational opportunities for the family, including good dining, the arts, music, hiking, wilderness, water sports and winter skiing nearby. This area is in a valley lying among the Adirondack, Catskill and Berkshire Mountains. Boston and New York City are 2-1/2 hour drives. Rochester, New York and Montreal are 4 hour drives. The area is noted for its many outstanding school districts.
Salary and benefits are competitive and commensurate with training and experience. AutoQuant is an equal opportunity employer, and will not discriminate based on race, color or national origin.
Please mail or fax your resume, a brief description of interests and the contact information for at least 3 references. If you respond by E-mail, please be sure to follow the E-mail with a regular mailing or fax. Please avoid encoded E-mail attachments. Respond to:
Human Resources AutoQuant Imaging, Inc. 1223 Peoples Ave. fax: 518-276-6380 Troy NY 12180-3536 E-mail: holmes-at-aqi3.aqi.com
It sounds like the Neurolucida software from MicroBrightField would handle your problem very well. You can get more information from them at info-at-microbrightfield.com or from their web site at www.microbrightfield.com/microb. You can also correspond with me; I know the system's details intimately.
On Thu, 22 May 1997, Heike Buecking wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I hope, that someone on the list can help me to find suitable PC-software } (DOS or Windows) or other tools for the following problem: } I have to reconstruct the three-dimensional structure of different } organs of microarthropods based on serial sections of resin embedded } specimen. Up to now I go the time consuming and not exact way of drawing } complete transverse, saggital and horizontal series and then to } "reconstruct" the 3D-structure more or less "by hand". Any idea is } welcome. } Heike } Dr. Heike Buecking } University of Bremen } UFT } Physiological Plant Anatomy } Leobener Str. } D 28359 Bremen } Germany } TEL: +49-421-218-2954 or } TEL: +49-421-218-7283 } FAX: +49-421-218-3737 } e-mail: heibueck-at-uft.uni-bremen.de }
Edmund Glaser, D. Eng. Dept. Physiol. Univ. Md. School. Med. Baltimore, MD 21201 USA Ph: (410) 706-5041 Fax: (410) 706-8341
Dear Colleagues, I have another question regarding to LR White: I am doing some immunostaining on semithin sections embedded in LR White. Although I know it is not absolutely necessary to remove LR White before the staining, I wonder whether there is an effective way to dissolve the cured resin but do not damage the tissue's fine structure. I faintly recall someone mentioned 70% alcohol can do the job. Is this correct? Is there other way which can be more effective? Thank you in advance for the help. Best regards, Yuhui Xu
I have been trying to locate a source for purchasing Lacomit Varnish and the Solvent and I haven't met with too much success. The one company I found does not have it in stock and has absolutely no ideal when they could get it. They said anywhere from now to maybe even next year. I have been waiting since last February and I think that is now long enough. Do any of you know where I can get it and is it in stock. I really don't want to place any more orders unless I know when I can get it. Thanks so much.
Peggy Bisher
NEC Research Institute 4 Independence Way Princeton, NJ 08540. Tel.: (609) 951-2629 Fax: (609) 951-2496 e-mail: peggy-at-research.nj.nec.com
Heike Buecking wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I hope, that someone on the list can help me to find suitable PC-software } (DOS or Windows) or other tools for the following problem: } I have to reconstruct the three-dimensional structure of different } organs of microarthropods based on serial sections of resin embedded } specimen. Up to now I go the time consuming and not exact way of drawing } complete transverse, saggital and horizontal series and then to } "reconstruct" the 3D-structure more or less "by hand". Any idea is } welcome. } Heike } Dr. Heike Buecking } University of Bremen } UFT } Physiological Plant Anatomy } Leobener Str. } D 28359 Bremen } Germany } TEL: +49-421-218-2954 or } TEL: +49-421-218-7283 } FAX: +49-421-218-3737 } e-mail: heibueck-at-uft.uni-bremen.de
We use the VoxBlast software to do just what you are asking for. It works well for us. You can get more information from the WWW site http://www.vaytek.com
Held alongside the CYTO 97 meeting, CRYO 97 promises to be a comprehensive update on the state of the art in Low Temperature Microscopy and Analysis. Come to York in July and enjoy a taste of both meetings and the large joint Trade Exhibition.
(this programme is easier read - I hope - if you maximise your email reader's window!)
CRYO 97 - Low Temperature Microscopy & Analysis - Programme
Sunday 6 July 12:00 - 14:00 REGISTRATION P.Monaghan Inst of Cancer Research, Sutton Introduction
SESSION A: SPECIMEN PREPARATION E Kellenberger Univ of Lausanne Retro-and perspectives of cryotechniques applied to sections: beliefs, dreams, misconceptions expectations, surprises and disappointments! M Miller To be advised. M Thijssen Wageningen Agricultural Univ Comparison of embedding fluids for high pressure freezing of Petunia ovules folled by freeze substitution. J Lepault CNRS, Gif surYvette Electron cryo-microscopy of biological tissues. D Studer Univ of Berne High pressure freezing of thick specimens.
Monday 7 July SESSION B: CRYO-MICROSCOPY H Saibil Birkbeck College, London To be advised. B Gowen EMBL, Heidelberg Visualising virus/vesicle fusion by time-resolved cryo-TEM. H Delacroix Univ P et M Curie, Gif sur Yvette Crystallographic analysis of freeze-fracture electron micrographs: application to the structure determination of cubic lipid systems. R Henderson MRC Lab of Mol. Biol., Cambridge Atomic resolution structure determination of biological macromolecules by electron microscopy. B Papahadjopoulos- Univ of the Pacific, USA Electron microscopic investigations of the fine-structure of lipid-coated DNA fibrils formed during Sternberg cationic liposomes/DNA interaction. N Atkin Univ of York Determination of salt-induced gellan polymer conformation by ultra-rapid freezing and low angle rotary shadowing. M van Heel Imperial College, London To be advised. D Studer Univ of Lausanne Electron beam induced changes in frozen hydrated specimens. S Butcher MRC Inst of Virology, Glasgow To be advised.
Tuesday 8 July SESSION C: CRYO-SEM W Jongebloed Univ of Groningen Cryo-preparation versus tannic acid/arginine/osmium tetroxide non-coating preparation observed with field emission SEM. J Nijsse Wageningen Agricultural Univ Cryo-SEM observations on frozen hydrated lettuce seeds during inhibition. K Robinson British Antarctic Survey Combination - To be confirmed. SESSION D: ELEMENTAL ANALYSIS G Roomans Univ of Uppsala Cryotechniques for preparation of tissue for X-ray microanalysis. P Echlin Univ of Cambridge Low temperature, low voltage microanalysis of diffusible elements in bulk frozen hydrated bio-organic samples. B Wolf Univ Freiburg Drug targeting and metabolic investigations on tumour cells with TEM-EELS analysis on cryo-prepared material. H Elder Univ of Glasgow Parameters for optimally freeze-drying cryosections of muscle for ultrastructural and microanalytical studies. A Pogorelov State Univ at Pushchino X-ray microanalysis of potassim deficit in cardiomyocyte induced by ischemia. I Karydis Univ of Cambridge Elemental changes presage apoptosis in human monocytes challenged with oxidised low density lipoprotein. SESSION E: APPLICATIONS L Edelmann Univ des Saarlandes Electron microscopy of freeze-dried biological material. I Lambrichts Limburgs Univ Centrum Cryomicroscopy of human periodontal ligament cells. N Hajibaheri ICRF, London High pressure frozen/freeze-substituted yeast and mammalian cells/tissue for immunocytochemistry. S Brookes Inst of Animal Health, Pirbright Cryo-electron microscopy of African swine fever virus: a novel pathway of virus morphogenesis as particles are wrapped by the Endoplasmic Reticulum. R Hermann ETH Zurich Structural and immunological studies of the nematode Heterorhabditis sp. and its host bacterium Photorhabdus luminescens, employing enhanced sampling techniques, followed by high pressure freezing, freeze substitution and plastic embedding.
POSTER PRESENTATIONS PC01 P Bennett Structural changes in samples cryo-fixed by contact with cold metal block. PC02 H Ekwall Cryo-electron microscopy of frozen boar semen PC03 B Martin Controlled freeze-drying for the preservation of fungal extracellular matrices PC04 E Shimoni A new micro biopsy system for high pressure freezing. PC05 P Wild Cryo-immobilisation and freeze-substitution of cell monolayers. PC06 C Winters A portable freezing and freeze-substitution assembly: anhydrous cryo-preparation in the field of a metal-binding freshwater moss for X-ray microanalysis. PC07 T Ishikawa Structural change of troponin induced by calcium ions are revealed by three dimensional cryo-electron microscopy. PC08 C Moores The interaction between utrophin N-terminus and F-actin: structural determination by cryo-electron microscopy. PC09 S Stoylova Structural of native PSII complex. A cryo-electron microscopy and crystallography study. PC10 G Daniel Use of HR-cryo-FE-SEM for studies on microbial degradation of wood and wood fibre structure. PC11 A Minnocci LTSEM and EDX X-ray microanalysis of wheat protoplast: a possible model for environmental stress investigations. PC12 A Minnocci LTSEM stomatal analysis in olive plants exposed to ozone. PC13 R Petacchi LTSEM sensilla studies on Dicyphus (D) erran (Wolff) (Heteroptera: Miridae). PC14 K Robinson An application of LT-SEM: how do hitch-hiking mites attach to coastal sand-hoppers? PC15 K Robinson Application of low temperature scanning electron microscopy: how eggs of a tardigrade respond to humidity changes. PC16 A Johnson Cryoprocessing for microanalysis of magnesium in the heart. PC17 A Morgan Hyperbaric freezing of invertebrate tissues: morphology and X-ray mapping of ion-transporting and metal-sequestering epithelia PC18 A Scully Microanalysis of freeze-substituted dentine materials. PC19 L Tetley The use of cryotechniques and energy filtering TEM for improved 3D cellular reconstruction: novel structure revealed in coccidian parasite infective stages. PC20 A Warley Freeze-drying and the preparation of cryosections for electron probe X-ray microanalysis. PC21 E Zellmann Cryo in-column EFTEM for free choice of detectors. PC22 D Carter Slam-frozen bone mineral. PC23 K Hovananyan Cryo-ultramicrotomy for electron microscopy study relation entamoeba with bacteria and liposomes. PC24 K Jennings Characteristics of beta amyloid peptide: a combined TEM, AFM and SEM study employing ambient and cryo-preparative techniques. PC25 C Milanesi New tips for freeze-substituting and immunogold labelling pollen grains from different species. PC26 N Perusinghe Ultrastructural immunocytochemistry of gap junctions in lactating mouse mammary gland. PC27 E L Punnonen Immunolabelling of 46 kDa mannose 6-phosphate receptor on cryo-sections using antibodies against the luminal domain and the cytoplamic tail. PC28 L Tetley Characterisation of polymeric glycol chitosan - a new drug delivery system - using cryo electron microscopy
Dr Laurence Tetley IBLS EM Centre Joseph Black Building University of Glasgow Glasgow G12 8QQ
We have a customer that is having difficulties with reembedding and would like some help. The following is what our customer wants to do: They want to reembed epon sections cut at 1-5 microns so that they can be resectioned at 60 nm. What is the best way to do this? The most effective with the least time and the fewest steps. Is thermanox the best material on which to collect these sections? Is there a mounting medium, or some other technique that will seal the sections to the Thermanox, but not create a thick layer on top of the section? Will the section sealed to the Thermanox actually cut at 60nm or will it just come off the Thermanox at some point? If the section on the Thermanox is inverted onto the top of a beem capsule fulled with unpolymerized epon and baked, how can the Thermanox be removed so that the section remains on the epon side? Will this procedure allow the section to remain flat and close to the surface during baking? Is there a reference that describes a useful protocol for this technique? I look forward to all responses.
} I have been trying to locate a source for purchasing Lacomit } Varnish and the Solvent and I haven't met with too much success. The one } company I found does not have it in stock and has absolutely no ideal when } they could get it. They said anywhere from now to maybe even next year. I } have been waiting since last February and I think that is now long enough. } Do any of you know where I can get it and is it in stock. I really don't } want to place any more orders unless I know when I can get it. Thanks so } much. } } } } Peggy Bisher
I would have expected most of the EM consumables suppliers to have - try SPI or Agar.
Dear all, We have had someone recently contact us regarding the use of a photographic-fixer rejuvenating service run by a company called Continu-Fix. I believe it is Canadian based. Essentially what is being offered is, we send our used film and paper fixes to this outfit, they will run it through some sort of refining process and then we buy back the fix as 'good as new' fixer. This then saves heaps of used fixer going down the drain and out into the environment, and saves us buying in new fix in 'un-environmentally friendly' bottles.
What we would like to hear from people regarding this is;
Is there any EM/LM/photo labs out there using this process or something similar? Do you believe that it is cost effective? Does the fix still have adequate fixing properties for archival quality? (ie; are we going to find that 6 months/6 years down the track, people will be coming back to us with black/brown prints).
We would appreciate any comments from anyone who has used this process,
Thanks in advance,
Rich.
----------------------------------------------------------------------- Richard Lander Senior Technician South Campus Electron Microscope Unit c/- Pathology Department Otago Medical School P.O. Box 913 Dunedin New Zealand. Tel. National 03 479 7301 Fax. National 03 479 7254
"Southernmost EM Unit in the World!" ------------------------------------------------------------------------
American Lab/News has provided an opportunity for us to review any new equipment/software/etc. which will be exhibited at the upcoming Microscopy & Microanalysis meeting. If you did not receive our fax, please contact our offices *immediately* and/or send words (50-75) and images (35 mm slides or prints) ASAP. We go to press next week.
Look forward to hearing from you. Barbara Foster, editor "Focus on Microscopy" c/o MME 53 Eton Street Springfield, MA 01108 (413)746-6931 fax: (413)746-9311 email: mme-at-map.com
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear colleagues,
A working group of the Dutch Society for Microscopy is collecting information on accreditation of microscopical work.
We would like to have information on:
1 Organisations that set formats for accreditation and certification in microscopy 2 certified standards for microscopy callibration 3 laboratories in microscopy that work under a accreditation regime yet 4 other interested parties
Thanks in advance,
Marcel Paques (chairman working group) Dutch Society for Microscopy
A working group of the Dutch Society for Microscopy is collecting information on accreditation of microscopical work.
We would like to have information on:
1 Organisations that set formats for accreditation and certification in microscopy 2 certified standards for microscopy callibration 3 laboratories in microscopy that work under a accreditation regime yet 4 other interested parties
Thanks in advance,
Marcel Paques (chairman working group) Dutch Society for Microscopy
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } It sounds like the Neurolucida software from MicroBrightField would } handle your problem very well. You can get more information from them } at info-at-microbrightfield.com or from their web site at } www.microbrightfield.com/microb. You can also correspond with me; I know } the system's details intimately. } } On Thu, 22 May 1997, Heike Buecking wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } -----------------------------------------------------------------------. } } } } I hope, that someone on the list can help me to find suitable PC-software } } (DOS or Windows) or other tools for the following problem: } } I have to reconstruct the three-dimensional structure of different } } organs of microarthropods based on serial sections of resin embedded } } specimen. Up to now I go the time consuming and not exact way of drawing } } complete transverse, saggital and horizontal series and then to } } "reconstruct" the 3D-structure more or less "by hand". Any idea is } } welcome. } } Heike } } Dr. Heike Buecking } } University of Bremen } } UFT } } Physiological Plant Anatomy } } Leobener Str. } } D 28359 Bremen } } Germany } } TEL: +49-421-218-2954 or } } TEL: +49-421-218-7283 } } FAX: +49-421-218-3737 } } e-mail: heibueck-at-uft.uni-bremen.de } } } } Edmund Glaser, D. Eng. } Dept. Physiol. } Univ. Md. School. Med. } Baltimore, MD 21201 USA } Ph: (410) 706-5041 } Fax: (410) 706-8341 } }
We have a customer that is having difficulties with reembedding and would like some help. The following is what our customer wants to do: They want to reembed epon sections cut at 1-5 microns so that they can be resectioned at 60 nm. What is the best way to do this? The most effective with the least time and the fewest steps. Is thermanox the best material on which to collect these sections? Is there a mounting medium, or some other technique that will seal the sections to the Thermanox, but not create a thick layer on top of the section? Will the section sealed to the Thermanox actually cut at 60nm or will it just come off the Thermanox at some point? If the section on the Thermanox is inverted onto the top of a beem capsule fulled with unpolymerized epon and baked, how can the Thermanox be removed so that the section remains on the epon side? Will this procedure allow the section to remain flat and close to the surface during baking? Is there a reference that describes a useful protocol for this technique? I look forward to all responses. Stacie Kirsch EMS
Hello all reembedders,
I have been reembedding 1-5 micron epon sections for years in the following way:
I section the tissue at 1-5 microns(the thicker the better) as if using for LM. That is I pick up the sections and place them on a drop of water on a glass slide. I heat the section as usual on a hot plate to afix it to the glass. After the slide cools off, I carefully scrape the section off the slide with a razor blade. I immediately pick the section up with tweezers and superglue the section onto a blank epon block. The blank block must be smooth, flat, and clean of course. I let the superglue dry well, and then the tissue is ready to be sectioned again. I have successfully then gotten ultra thin sections in the usual way.
Others in our lab have done the above, except they use epon itself rather than superglue to afix the 1-5 micron section to the blank block. I don't like this method because it takes longer ( you have to put the block combo back in the oven to polymerize ). This method is also good for those more patient than I.
Hope this works well for you. Hope I didn't leave anything out of the technique.
Sections can be collected on untreated glass microscope slides, dried on a hotplate, stained with toluidine blue if desired, then covered with an inverted BEEM capsule (the small ones work best) filled with unpolymerized resin. Place them in a 90C oven for 60 - 90 minutes, then peel the blocks off while the blocks and slides are still hot. If the slides cool off, put them back into the oven for 5 - 10 minutes and try to peel the blocks off again. The blocks may need a little more time in the oven to fully polymerize after they've been removed from the slides.
The tricks are to use plain microscope slides (no poly-L-lysine, silanes, or acid cleaning) and to remove the blocks while the plastic is still slightly soft.
Jane A. Fagerland, Ph.D. Dept. Microscopy and Microanalysis Abbott Laboratories Abbott Park IL 60064
We have routinely used parlodion coated grids for our EM thin sections. 2.5% parlodion in amyl acetate dropped on to distilled water, then lowered on to the grids and dried. However, lately we are having problems with uneven thickness. We have tried new amyl acetate and different percentages of parlodion but still no luck on solving the uneven thickness.
Any ideas?! Or protocols that work for you?
Thanks in advance for any help.
Bob Underwood Morphology Core U of Washington Seattle, USA
If I am not mistaken, a lab on the floor above us has the Polaroid Sprintscan (it is definitely a Polaroid, whether it is the same model Sprintscan I don;t know). The resolution is fine. It is easy to use. It does not advance the film automatically. It is slow at 2700 DPI, but otherwise decent speed.
It only take 35 mm or smaller, so it does not meet your specified needs. It can be used, if you cut a custom size metal insert or buy one from a company that has been advertising heavily (but I forgot the name anyway) to scan microscope slide.
-------------------------------------------- email sent from an account of the Analytical Imaging Facility The Albert Einstein College of Medicine of Yeshiva University 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 FAX: (718) 430-8996 --------------------------------------------
The following message which was sent by Analytical Imaging Facility {aif-at-telico.bioc.aecom.yu.edu} had incomplete information concerning Polaroid scanners, so I thought I'd respond even though I have no financial interest in Polaroid. The SprintScan 45 is a multi-format film scanner which scans film as large as 4x5 inch to 35 mm format. The SprintScan 35 is a scanner for 35 mm format films only. See Polaroid's web page at http://www.polaroid.com/products/scanners/index.html. } } If I am not mistaken, a lab on the floor above us has the Polaroid } Sprintscan (it is definitely a Polaroid, whether it is the same model } Sprintscan I don;t know). The resolution is fine. It is easy to use. } It does not advance the film automatically. It is slow at 2700 DPI, but } otherwise decent speed. } } It only take 35 mm or smaller, so it does not meet your specified needs. } It can be used, if you cut a custom size metal insert or buy one from } a company that has been advertising heavily (but I forgot the name } anyway) to scan microscope slide. } } -------------------------------------------- } email sent from an account of the Analytical Imaging Facility } The Albert Einstein College of Medicine of Yeshiva University } 1300 Morris Park Ave. Bronx, NY 10461 } (718) 430-2890 FAX: (718) 430-8996 } --------------------------------------------
Russell E. Cook Scientific Associate Electron Microscopy Center for Materials Research Argonne National Laboratory Building 212 9700 South Cass Avenue Argonne, IL 60439 (630)252-7194 FAX: (630)252-4798
} We have routinely used parlodion coated grids for our EM thin sections. } 2.5% parlodion in amyl acetate dropped on to distilled water, then lowered } on to the grids and dried. However, lately we are having problems with } uneven thickness. We have tried new amyl acetate and different } percentages of parlodion but still no luck on solving the uneven } thickness.
Several causes of uneven thickness in Parlodion (Collodion, cellulose nitrate, etc) are:
1. too rapid evaporation rate of the solvent carrying the parlodion on the water surface (possibly due to the water being too warm, or the room too warm, or air currents - as from a fume hood - blowing over the water surface),
2. incomplete dissolution of the parlodion in the amyl acetate (possibly due to not waiting long enough to dissolve, or slightly moist parlodion flakes/chips),
3. the parlodion solution and the water should be the same temperature (20-24 deg C).
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 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 ####################################################################
Marcel Paques wrote: ========================================== } A working group of the Dutch Society for Microscopy is collecting } information on accreditation of microscopical work. } } We would like to have information on: } } 1 Organisations that set formats for accreditation and certification
} in microscopy } 2 certified standards for microscopy callibration } 3 laboratories in microscopy that work under a accreditation regime yet } 4 other interested parties ================================================ In the USA, the American Association for Laboratory Accreditation (A2LA) has been accrediting laboratories in "microscopy" under the "chemical" discipline. The accreditation in microscopy started in 1980 and today there are some number of laboratories accredited by A2LA and also, the accreditation today is being done to the standard of ISO Guide 25.
For SEM calibration, there is the NIST Certified Reference Material. But for TEM it is more complicated, and what accredited laboratories end up using seems to be determined in part by the preferences of the one doing the assessing. The two main approaches are either a) the use of a diffraction grating from which one figures out the precise number of lines from the optical properties and b) dimensional samples, such as calibrated spheres that are reported traceable to NIST.
Are there laboratories actually operating under this accreditation regimen? Absolutely! Are there very many? Not too many. An up-to-date list can be obtained through A2LA:
American Association for Laboratory Accreditation 656 Quince Orchard Rd. #620 Gaithersburg, MD 20878-1409 301-670-1377, FAX 301-869-1495 http://www.a2la.org/ E-mail: You can contact them through their website above
Internested parties? Other than A2LA above, ACIL would certainly fall into that category. You can reach ACIL at {http://www.acil.org/} , which is the professional and trade association of independent testing, analytical and engineering laboratories in the USA.
One might ask whether this program "works". Yes, it does work and it works very well. And I speak from first hand experience since the laboratory part of our business has been accredited by A2LA since the inception of their program in 1980.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I am collecting information on the currently valid standards applied in metallographic examinations in different countries and corresponding to the following standards of ASTM;
A247 - 67 Test Method for Evaluating the Microstructure of Graphite in Iron Casting
E3 - 95 Preparation of Metallographic Specimens
E7 - 96 Metallography
E45 - 95a Determining the Inclusion Content of Steel
E1245 - 95 Determining the Inclusion or Second-Phase Constituent Content of Metals by Automatic Image Analysis
E1268 - 94 Assessing the Degree of Banding or Orientation of Microstructures
E1382 - 91 Determining Average Grain Size Using Semiautomatic and Automatic Image Analysis
E1558 - 93 Electrolytic Polishing of Metallographic Specimens
ISO 9042 - 88 Steels: manual point counting method for statistically estimating the volume fraction of a constituent with a point grid.
JIS G 0555 - 77 Microscopic Testing Method for the Non-metallic Inclusions in Steel
If such information is available kindly send me the data arranged in the following sequence; No. of standards according to ASTM, ISO, JIS - number of the corresponding national standard, its title (English and national), date of publication and the date of last revising.
If you think that in the list given above someother relevant standards have been omitted, kindly let me know their numbers and titles.
Thanking you in advance for your kind co-operation and assistance, I remain
yours sincerely
Krzysztof Jan Huebner
{hubner-at-IOd.krakow.pl} :-)
FOUNDRY RESEARCH INSTITUTE Research Materials Department Structural and Physical Research Laboratory
Recently there was discussion of Aclar film and its myriad uses, including reembedding thick sections. I recently bought some from ProSciTech (haven't used it yet) and it was said to be available from Electron Microscopy Sciences. From memory, the Aclar is cut into slide size pieces and used instead of an ordinary slide. The section is stained as usual, then inverted over an embedding mould with resin and cured. The Aclar can be sectioned, is light and electron lucent and is chemically inert. The ProSciTech web page has some info and references about Aclar: http://www.proscitech.com.au/l35.htm#l105. Diana
Diana van Driel Dept Ophthalmology Sydney University C09 AUSTRALIA 2006
Here are some references which describe the methods for re-sectioning semi-thin (or thick) resin sections. Our procedure included : (1) prepare 4um thick sections with a histo diamond knife and mount on microscope slides and stain with toluidine blue. (2) stick re-faced resin blocks on the mounted sections with a drop of epon-araldite or crazy glue. (3) leave in oven for 2 days at 60C (4) "pop off" the blocks after leaving on dry ice for 45 minutes.
References : Di Sant'Agnese & De Mesy-Jansen (1984). Ultrastruct. Pathol 6: 247-253
Milroy (1985). J. Electron Microsc. Tech. 2: 399-400
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } We have a customer that is having difficulties with reembedding and would } like some help. The following is what our customer wants to do: } They want to reembed epon sections cut at 1-5 microns so that they can be } resectioned at 60 nm. } What is the best way to do this? The most effective with the least time and } the fewest steps. } Is thermanox the best material on which to collect these sections? } Is there a mounting medium, or some other technique that will seal the } sections to the Thermanox, but not create a thick layer on top of the } section? } Will the section sealed to the Thermanox actually cut at 60nm or will it just } come off the Thermanox at some point? } If the section on the Thermanox is inverted onto the top of a beem capsule } fulled with unpolymerized epon and baked, how can the Thermanox be removed so } that the section remains on the epon side? Will this procedure allow the } section to remain flat and close to the surface during baking? } Is there a reference that describes a useful protocol for this technique? } I look forward to all responses. } } Stacie Kirsch } EMS }
We are interested in adding a 2k x 2k image digitiser to a non digital SEM. If you know of such a system, could you advise us of the supplier, and, preferably, the price? 1k x1k systems are not appropriate.
thanks,
Mel Dickson Mel Dickson President, Australian Society for Electron Microscopy Director, Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
We have a JEOL 200CX which is currently running on Freon. Are there any points or pitfalls which we should know about before we undertake a conversion.
Thanks in advance.
Regards,
Keith. ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Dr. K. Moulding.
Materials Characterisation and Preparation Facility Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong.
Dear Mel, I have the Quartz PCI system, which captures images up to 4096 X 4096, depending upon the output of the SEM. They can hook up to any old SEM and uses a Pentium computer for display. It costs about $15,000CAN. In Canada the supplier is NSC, the Hitachi microscope supplier. You can contact the Quartz company at adbrown-at-qrtz.com (Andrew Brown) to ask about Australian suppliers. The lastest version does lots of neat things. You wrote: } We are interested in adding a 2k x 2k image digitiser to a non digital SEM. } If you know of such a system, could you advise us of the supplier, and, } preferably, the price? 1k x1k systems are not appropriate. } } thanks, } } Mel Dickson
Regards, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
I believe that ELI, a company here in Belgium, has what you are looking for. They have a very good product for your application and they are reliable. I believe that they sell worldwide and you can contact them for more info via E-mail at: orion-at-infoboard.be
The Sales Mgr is Paul Vanderlinden.
Disclaimer: I am not affiliated nor have any interest with that company, I am just a satisfied customer of one of their products.
Good luck and best regards, Michel
**************************************************** Michel Deschuyteneer, Ph.D. deschuyt-at-sbbio.be Scientist Electron Microscopy Laboratory
SmithKline Beecham Biologicals Rue de l'Institut, 89 B1330 Rixensart, BELGIUM Tel: +32-2-656 9290 Fax: +32-2-656 8164 **************************************************** Standard disclaimer: the opinions expressed in this communication are my own and do not necessarily reflect those of SmithKline Beecham. ****************************************************
There is a very good, reliable, well designed DISS system from German company Point Electronic. System consists from one specially designed PCB installed in standard PC with enough configuration. Can scan with adjustable resolutions from 64x64 till 4096x4096 pixels. It is mains-locked and stops on each scan point integrating signal. PCB has 8 video inputs and 6 digital TTL inputs for xray maps etc. allowing simultanous aquisition. Digitizing goes with 12 bit a/d converter. The newest software is very clever design and runs under windows as TWAIN driver - so for image manipulation one uses any PhotoStyler, Shop, etc software and selects for Aquire-command the SEM driver - it works excellent. Price range fro system is below 30.000 DM but includes PC, instalation (interfacing to SEM) etc. - in case of Australia will be probably different. There are few hundreds of such systems allready running around Europe. - contact is Dr W.Joachimi Point ElectronicGmbH Koethener Str 34, 06118 Halle/Saale, Germany phone: (+49 345) 202 3996 fx: (+49 345) 4235254
kind regards Krzysztof M. Herman (we are polish distributor of Point Electronic) LabSoft S.c. 05-500 Piaseczno, 21 Kosciuszki Str. Poland tel/fx: (48 22) 7502024, 7502028, 7570671 fax only: (48 22) 483787, Email: labsoft-at-ikp.atm.com.pl
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
My method is very similar to that used by Jane. When you have found the section of choice cut the slide into small pieces and lay the piece, with the section, on top of a Beem capsule slightly overfilled with resin then polymezire. Remove the block from the Beem capsule, briefly dip the glass into liquid nitrogen which will then fall away leaving block with section attached. Ian.
Wanting to control the presence of esterified pectins in plant cells I've tried the hydroxyalmine/ferric chloride reaction as described by Reeve (1959). I suppose this reaction is routinely used by many of us but.... I have a precipitation problem when I add to the fresh hydroxylamine reagent - i.e. a solution of equal volumes of sodium hydroxide (14g in 100ml EtOH 60%) and hydroxylamine hydrochloride (14g in 100ml EtOH 60%) - concentrated HCl in EtOH in order to acidify the reaction mixture. The precipitate I obtain is of cristalline structure and rather sticky... If you met this problem to when following exactly Reeve's protocole, if you were lucky in solving this problem or if you were not, etc... Please help ! Thanks Pascal
************************************ Pascal VEYS Laboratory of Plant Biology Catholic University of Louvain Place Croix du Sud 5 (bte 14) B 1348 Louvain-la-Neuve Belgium Phone : 0032 10473004 Fax : 0032 10473471 Email : Veys-at-bota.ucl.ac.be ************************************
In a message dated 97-05-26 23:39:41 EDT, M.Dickson-at-unsw.edu.au (Melvyn Dickson) writes:
} We are interested in adding a 2k x 2k image digitiser to a non digital SEM. } If you know of such a system, could you advise us of the supplier, and, } preferably, the price? 1k x1k systems are not appropriate. } }
You might want to give EOS and/or Evex Analytical (609-252-9192) a call. I know Evex has put together a harware & software package including Image Pro. The hardware is 4k by 4k, 12 bit, active imaging system. I believe the PCI system is passive
The price is $9,000 to $20,0000 depending on system configuration.
} There is a very good, reliable, well designed DISS system from German } company Point Electronic.
In the interests of not boring the rest of the subscribers to tears, I thought we vendor types were going to refrain from posting spec sheets to the whole list in response to requests for information.
Let's agree to send this kind of post just to the person making the request, please, and the list will be more enjoyable and useful for all of us...
Hello from Polaroid -- I am Brooks Corl, Senior Applications Manager.
You are clearly describing the Polaroid "SPRINTSCAN 35" scanner, which scans 35mm transparencies or negatives. There is also another model, "SPRINTSCAN 45", that scans up to 4x5 transparencies or negatives. That was the model, I think, that the original question concerned.
Please check our web site (www.polaroid.com) for more information, or feel free to get back directly to me via cc-mail (corlb-at-polaroid.com) if you have questions I can answer or research for you about these scanners. Thanks!
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
If I am not mistaken, a lab on the floor above us has the Polaroid Sprintscan (it is definitely a Polaroid, whether it is the same model Sprintscan I don;t know). The resolution is fine. It is easy to use. It does not advance the film automatically. It is slow at 2700 DPI, but otherwise decent speed.
It only take 35 mm or smaller, so it does not meet your specified needs. It can be used, if you cut a custom size metal insert or buy one from a company that has been advertising heavily (but I forgot the name anyway) to scan microscope slide.
-------------------------------------------- email sent from an account of the Analytical Imaging Facility The Albert Einstein College of Medicine of Yeshiva University 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 FAX: (718) 430-8996 --------------------------------------------
This may be too obvious, but did you make sure the surface of the water that you are dropping the parlodion onto is as level as possible? Uneven thickness occurs if the surface is concave or convex. If the uneven thickness is random or patchy, you probably have a contamination problem.
Karen Pawlowski
On Fri, 23 May 1997, Robert Underwood wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello fellow microscopists, } } We have routinely used parlodion coated grids for our EM thin sections. } 2.5% parlodion in amyl acetate dropped on to distilled water, then lowered } on to the grids and dried. However, lately we are having problems with } uneven thickness. We have tried new amyl acetate and different } percentages of parlodion but still no luck on solving the uneven } thickness. } } Any ideas?! Or protocols that work for you? } } Thanks in advance for any help. } } Bob Underwood } Morphology Core } U of Washington } Seattle, USA } } }
Gatan offers DigiScan, a system that converts your analog SEM to Digital. It will digitize up to 8k x 8k and will do more than that, it will drive the scan coils of your SEM. It is versatile and user friendly. The PCI system is a passive capture system and will not give control of the instrument. We have recently installed DigiScan on our 20 year old AmRay 1000A and it drives like a new microscope!
Gatan has a website which provides information and a demo copy of their micrograph image processing software, Digital Micrograph.
Disclaimer: We working with Gatan on developing remote microscopy but have no financial interest in the product or company.
Jacob
Jacob Bastacky, M.D. Room 116 Donner Laboratory Lawrence Berkeley Laboratory EMail: sjbastacky-at-lbl.gov University of California Phone: (510) 486-4606 Berkeley, California 94720 Fax: (510) 486-4750
Is there an imaging system or a program specifically made for bone morphometry? We are using a regular multipurpose one presently , but it takes long time and certain measurements we still have to do by hand. Thanking you in advance,
WE are sending, via federal express for am delivery, which should arrive at your offices in Springfield by Wednesday May 28, 1997, a photograph and descriptive copy for our Tacky Dot (tm) slides.
Please include this information in your review of new products to be published in the Microscopy & Microanalysis issue of American Lab/News.
THANKS for The Opportunity to expose our products to your audience!!!
I'm trying to locate David J. Barber, recently at the Physics Dept., Hong Kong Univ of Sci and Tech. I tried his e-mail there and it wasn't delivered. He isn't listed on their www site.
If any one knows where he is please send me a note off line.
Medjet is a public compamy for Refractive Surgery Instruments. Need to hire a person with histology skills (light microscopy, scanning microscopy), computer skills and experience in animal experiments; full-time job. Our phone is (908) 7383990; our fax is (980) 7383984
First, thanks to all of you who responded with suggestions on how to fix the leak in my nitrogen dewar. Most of the responses suggested I use a silicone sealant or an epoxy compound.
I do have a follow-up question though. Any suggestions on what's the best way to remove the flange ?. I think that in order to properly fix the leak I should first remove the flange from the dewar (rather than just applying an epoxy or sealant on the surface). The way I understand it, the bottom part of the flange fits into the dewar and an epoxy is used to seal it in place. In my case the leak seems to be right in this epoxy seal. Is there a proper way to remove this flange without damaging the rest of the flange or the dewar ? I don't think that brute force will do it, but what about heat ? If I heat up the neck of the dewar to burn off the epoxy, will I be damaging insulation in the dewar or in the flange ?
German overseas newscast reported the death of Manfred von Ardenne today. During the '30s he was involved in cathode ray work and later lived in East Germany and did significant cancer research. Not mentioned was his conceptual development of the SEM in the late 1930s. He outlined the underlying principles of SEM operation: an electron probe scanning a small region of the specimen, the emitted electrons are captured, amplified and time-sequentially displayed on a cathode ray tube. Magnification is the ratio between the area scanned and displayed. His was the fantastic notion of a microscope without a magnifying lens. It is interesting to contemplate that the now more complex TEM was built and developed from the mid '30s and was a high performer in the early '60s. It was at that time when the Cambridge group, under Oatley, was able to build the first SEM. A lot of technology had to mature before SEM was possible and the Cambridge group deserves great credit. But the development of several unique concepts of a then quite futuristic instrument is to von Ardenne's enduring credit. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 350+ Links, MSDS ************************ http://www.proscitech.com.au
Dear Mr Mott and all vendors/distributors who trace the user's discussions OK about this.... no problem for future ....but why to read boring things and cry? ... just click delete message like by SUBSCRIBE / UNSUBSCRIBE ones!! the real mess would be to give NON REQUESTED ADVERTISING which we not do. regards Krzysztof M. Herman LabSoft S.c. 05-500 Piaseczno, 21 Kosciuszki Str. tel/fx: (48 22) 7502024, 7502028, 7570671 fax only: (48 22) 483787, Email: labsoft-at-ikp.atm.com.pl
I'd like to chime in on another development of Manfred von Ardenne.
Back in my foundry days, I ran a marvelously engineered piece of machinery; an 800kW electron beam furnace. The heart of this machine was array of independently-controlled beams coming from four von Ardenne-produced guns. The furnace was five stories tall and could melt and purify a ton of tantalum an hour. Absolutely amazing technology.
------------------------------------------------ Opinions or statements expressed herein, rational or otherwise, do not necessarily reflect those of my employer.
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 Phone: (508) 750-1717 E-mail: crossman-at-osi.sylvania.com
Our web sites: www.sylvania.com www.siemens.com --
My wife has finished her class and should be getting her high school certification soon, provided that the rules don't change again. Up to this point, I have compiled the answers we've received from the Microscopy server. This was by far the richest source of responses. I would be happy to provide a copy of the responses from the server to anyone who is interested.
Attached is my wife's thank you. Thank you all very much for your help.
Chuck
P.S.
Dear Answer to a Prayer Scientists,
Thanks for taking the time to answer my survey. In all we had 17 responses from scientists in all walks of life. Many of you had some of the same ideas with slightly different twists. For the most part, people felt the history of science was important in education and research. The ideal classroom was a hands on experimental setting. The nature of science responses were varied just as it is in educational circles. Once again, thanks. Debbie Gilbert
------------------------------------- Name: Charles Gilbert VOC:(704)355-5261 Carolinas Medical Center FAX:(704)355-8424 Dept of Pediatric Research PO Box 32861 Charlotte, NC 28232-2861
Is it possible to do negative staining (2% PTA) on a suspension of virus -- which has been fixed with 1% glut. in cacodylate buffer? In the past I've only used unfixed materials. Thanx.
John Hardy E.M. Lab City of Hope Medical Center jhardy-at-smtplink.coh.org
First I want to thank everybody who responded to my question on suitable software to reconstruct the 3D-structure of organs (based on serial sections). I think it's a good idea to summarize all the tips I have got the last week. The list below is far from beeing complete (sorry to everybody who thinks that I have forgot something important).
NIH-IMAGE This popular freeware programme for image analyses is originally written for the Mac, but now is also available as Win95 program. Download: http://www.zippy.nimh.nih.gov/ (Mac)or http://www.scioncorp.com/ (Win95) Many informations e.g. online manual, macros, example-files and additional download possibilities can be found at: http://rsb.info.nih.gov/nih-image/ As an example animated reconstructions of plant cells (based on semithin sections) can be found on the homepage of Gary Chinga: http://www.nvg.unit.no/~gary Informations about the NIH-Image mailing list can be found at http://www.soils.agri.umn.edu/infoserv/lists/nih-image/
NEUROLUCIDA Informations about Neurolucida are available at http://www.microbrightfield.com/microb Although Neurolucida is primarily designed for the needs of neurobiologists, it has all the facilities which I need for the planed reconstruction based on serial tissue sections (Data entry by camera or digitizing tablet, alignment, tracing of areas of interest, 3D-modelling and volume measuring).
VOXBLAST Voxblast is available for Unix, Mac and Windows-Sytems. Informations about this software, e.g. instructive demo-versions and a quick-reference-quide can be found at http:/www.vaytek.com/ In order to transfer and prepare (enhancement and alignment of the slices) the images for the use with Voxblast, suitable image analyses software, e.g. NIH-image or ImageProPlus from Mediacybernetics (http://www.mediacy.com/) is needed.
T3D (formerly SLICER) Included in the data-management-package NOESYS of Fortner research. Product informations can be found at http://www.fortner.com/
SLICER DICER Informations about this volumetric data visualization software of Visalogic (available for MAC, Win95 and WinNT) could be found at http://www.visualogic.com/
MONTAGE This 3D reconstruction package is a UNIX-programm, but works also on a PC using the free Unix-system LINUX. It's available by anonymous FTP at ftp://retina.anatomy.upenn.edu:/pub/mont.linux.tgz Data entry is simply by a Summagraphics Digitizing Tablet.
A list of links to the distributors of the above mentioned software and many other programs for image analyses can be found at: http://ddsdx.uthscsa.edu/dig/sites.html
This is all I get up to now from the internet and of course I haven't tested all the programs listed above for their suitability.
Sincerely,
Jens Buecking
--------------------------------------------------------------- Dr. Jens Buecking Tel.: +49-(0)421-218 3745 University of Bremen Fax.: +49-(0)421-218 4042 Department of Biology email: jbueck-at-biologie.uni-bremen.de Leobener Str. - NW2 D- 28359 Bremen Germany ---------------------------------------------------------------
Dr. Heike Buecking University of Bremen UFT Physiological Plant Anatomy Leobener Str. D 28359 Bremen Germany TEL: +49-421-218-2954 or TEL: +49-421-218-7283 FAX: +49-421-218-3737 e-mail: heibueck-at-uft.uni-bremen.de
One of the scientists here wishes to use one micron gold particles as carriers for a plant virus in some transmission studies. He has asked me for suggestions re how to bind the virus to the gold. I don't have any references dealing with this size of gold particle, but have several questions: Can the virus be directly bound to these large gold particles? How? Could the specific antibodies be bound to the gold (as with smaller colloidal gold)? Is there perhaps a commercial supplier of one micron gold conjugated to an anti-rabbit antibody? Or something that will bind a virus?
I am currently looking at catepillars by SEM and would like to do TEM. Does anyone know of a specific stain that could be used prior to embedment to make the nerve cells in the taste receptors stand out? Is there a specific stain that could be used after embedment rather than using UA and PbCitrate? Thanks in advance.
Peace through Christ,
Phil Rutledge, Director e-mail: prutle1-at-gl.umbc.edu Center for Microscopy voice: (410) 455-3582 UMBC Dept. of Biology fax: (410) 455-3875 Catonsville, MD 21228 ///// / / / / /////// /////// / / /////// /////// / / / / / / / / / / / / /////
} Is it possible to do negative staining (2% PTA) on a suspension of } virus -- which has been fixed with 1% glut. in cacodylate buffer? In } the past I've only used unfixed materials. Thanx. }
Yes, this can be done with the following caveats:
1. If the virus was fixed while in suspension (not pelletized or in tissue) you should be OK otherwise the virus will be cross-linked to other virus or tissues in an inseparable pastiche that the beam will not penetrate.
2. The glut/cacodylate should be washed out, if possible, to prevent salt crystals from forming during the drying of the grid. An easy way of doing this would be to float the coated grid first on a suspension of the fixed virus in the glut/cacodylate for several minutes followed by floating the grid on 2% PTA for several seconds.
It certainly wouldn't take much to check it out.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
In message {9704288648.AA864839277-at-smtplink.coh.org} "HARDY, JOHN" writes: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Is it possible to do negative staining (2% PTA) on a suspension of } virus -- which has been fixed with 1% glut. in cacodylate buffer? In } the past I've only used unfixed materials. Thanx. } } John Hardy } E.M. Lab } City of Hope Medical Center } jhardy-at-smtplink.coh.org
Give it a try, but before you set up the neg stain, pellet the virus in a microfuge or ultracentrifuge (whatever it takes to get you enough g's), discard fixative supernatant, and resuspend either in distilled water or very weak buffer, ie, about 0.01 to 0.05 molarity. Otherwise the glut and the buffer will generally clog up the prep., coat the virus and mess things up.
Good luck!
--
Gib Ahlstrand, MMS Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
I have a questions which has been asked of me in the past: Why is it that there is less charging in a Field emission electron microscope as oppossed to a standard LaB6 or W system?
At about 1keV for most insulators, there is a charge balance condition between the number of electrons coming in (incident current) and the number of backscattered and secondary electrons (backscattered and secondary currents) so that I0= Iback + Isec. The absorbed current is zero (that's why it is an insulator). You can operate a FEG-SEM at lower voltages and get more electrons than with either LaB6 or W thermionic sources. The current available in these decreases with decreasing voltage. With a W source, there just isn't enough current density giving a sufficient signal to noise ratio to do anything. It gets better with a LaB6 and there's plenty with a FEG.
The answer to your question is that there isn't less in a FEG. The charging is dependent on the operating voltage of the microscope. If the voltages are the same in the different microscopes, then the above equation holds. In the W and LaB6 machines, you have to crank up the voltages to see anything in the image and you no longer have the charge balance condition. You can increase the voltage that you can work at by tilting the sample to high angles. For example if the insulators have charge balance at 1 kV 0 deg tilt, then the balance condition will be about 3 kV at 70 deg tilt. This condition can be used in an scanning Auger microscope for insulators if a orthoganal detector is used. (JEOL SAM's)
The question that I have about charging in an SEM is this. If you set up the imaging conditions for the sample not to charge at a high scan rate i.e. TV rate, and then drop it to a slow scan (as you do in an analog machine when recording an image), why does the image now charge?
- -
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I have a questions which has been asked of me in the past: } Why is it that there is less charging in a Field emission electron } microscope as opposed to a standard LaB6 or W system? } } ...
I can think of only two reasons ... (1) FE guns acquire noise-free images at lower beam currents ... or, (2) FE guns can't deliver higher beam currents. However, conventional guns at the same beam current shouldn't charge more ...
cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
How easily can bone be thin sectioned without demineralization? We are interested in viewing the callus area and osteoblasts and want to preserve this area as best as possible. I would appreciate any information or referral to literature. Thanks. Bill Meek, Ph.D. Meekwd-at-okway.okstate.edu
} I am currently looking at catepillars by SEM and would like to do TEM. } Does anyone know of a specific stain that could be used prior to embedment } to make the nerve cells in the taste receptors stand out? Is there a } specific stain that could be used after embedment rather than using UA and } PbCitrate? Thanks in advance. } } Phil Rutledge
Phil,
In brief, I think the answer is "no". When I was looking at amphipod sensory structures, the cells had to be identified by location, structure, and tracing contacts. LaNO3 could be used to detect intercellular spaces and tight junctions (the LaNO3 was supposed to not diffuse past them), but no specific stains for neurons.
Having said that, you might make a go of it by labelling the nerves, or receptor end-organs with horseradish peroxidase and staining by the usual methods. This requires keeping the beasties alive for a few days to let the HRP diffuse. A steady hand, no coffee, and minute dissecting instruments help.
If they're all dead, you might try the Di-I and Di-O fluorescence methods used in tracing mammal and bird nerves. This would give you a LM image of where the taste bud neurons are, and they could then be sought in the TEM. If it works. I had trouble using these on fish, and as I recall, they didn't work well aside from mammals and birds.
This could all be outdated now, and I would be happy to be corrected!
Phil
} Sic Hoc Legere Scis Nimium Eruditionis Habes { Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217) 355-1143 oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
[Apologies if this appears twice, my email program hiccuped.) } I am currently looking at catepillars by SEM and would like to do TEM. } Does anyone know of a specific stain that could be used prior to embedment } to make the nerve cells in the taste receptors stand out? Is there a } specific stain that could be used after embedment rather than using UA and } PbCitrate? Thanks in advance. } } Phil Rutledge
Phil,
In brief, I think the answer is "no". When I was looking at amphipod sensory structures, the cells had to be identified by location, structure, and tracing contacts. LaNO3 could be used to detect intercellular spaces and tight junctions (the LaNO3 was supposed to not diffuse past them), but no specific stains for neurons.
Having said that, you might make a go of it by labelling the nerves, or receptor end-organs with horseradish peroxidase and staining by the usual methods. This requires keeping the beasties alive for a few days to let the HRP diffuse. A steady hand, no coffee, and minute dissecting instruments help.
If they're all dead, you might try the Di-I and Di-O fluorescence methods used in tracing mammal and bird nerves. This would give you a LM image of where the taste bud neurons are, and they could then be sought in the TEM. If it works. I had trouble using these on fish, and as I recall, they didn't work well aside from mammals and birds.
This could all be outdated now, and I would be happy to be corrected!
Phil
} Sic Hoc Legere Scis Nimium Eruditionis Habes { Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217) 355-1143 oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
Hello again, Requesting help this time on the repair of our Hitachi H-600 film exchange. The films jam by not falling totally into the slot. When the air compressor moves them forward after the exposure, the front left corner of the plate jams on the support and is wedged in place. The entire camera must be opened, I reach inside (in the dark) and unjam the film. The assembly snaps back into place and seems ok for two or more films and then does it again. Our service contract was not renewed by our sage management team, and Hitachi has not yet returned a call on help over the phone, if help this way is available to us at all. Our in house PP&G people want to use the big hammer and lots of lubricant. HELP Linda Fox lfox1-at-wpo.it.luc.edu
} The question that I have about charging in an SEM is this. If you set up the } imaging conditions for the sample not to charge at a high scan rate i.e. TV } rate, and then drop it to a slow scan (as you do in an analog machine when } recording an image), why does the image now charge?
Because in a slower scan mode the beam dwells longer on the specimen permitting more of a static charge to build up; the charge collects with time.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 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 ####################################################################
Generally, FESEMs are operated at an accelerating voltage of around 1kV. At this voltage, most none conductive samples will not charge. Similar charging characteristics will arise at higher voltages regardless of electron gun type.
Steve Joens Standard SEM Section Manager Hitachi Scientific Instruments JOENS_S-at-NISSEI.COM
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have a questions which has been asked of me in the past: Why is it that there is less charging in a Field emission electron microscope as oppossed to a standard LaB6 or W system?
X-Sender: awilson-aw-at-mail.u-net.com (Unverified) X-Mailer: Windows Eudora Light Version 1.5.2 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii" To: Microscopy-at-Sparc5.Microscopy.Com
ATTENTION ALL IMMUNOHISTOCHEMISTS, IMMUNOCYTOCHEMISTS AND OTHER AFFINITY LABELLERS!!! VOTING IS NOW IN PROGRESS FOR A NEW IMMUNOCYTOCHEMISTRY NEWSGROUP!
The official CFV or CALL FOR VOTES for the proposed new newsgroup sci.bio.immunocytochem has recently been posted to news.announce.newgroups and news.groups. If you are keen to see a newsgroup dedicated to discussion of immunocytochemistry and related topics, then please do vote. All you need to qualify as a voter is a valid e-mail address.
To vote for the new group, use your newsreader to access USENET, find {news.announce.newgroups} or {news.groups} and look for the official posting named {CFV:sci.bio.immunocytochem} (posted on 26th May 1997) The VOTING INSTRUCTIONS and the BALLOT can be found at the end of the CFV. You MUST use the official ballot to vote, and it MUST be posted to the official votetaker for it to be counted. Thank-you.
Amanda Wilson, e-mail {awilson-at-aw.u-net.com} Proponent of the proposed newsgroup {sci.bio.immunocytochem}
Well, I'm about to leave Oz and EM to move to Europe and hopefully start a Ph.D. in Archaeology (a long way from Marine Zool. and EM!) but before I go I've got to do an animated sequence from the SEM, in stereo, on a very small spider (a couple of thousand frames). Should look good. The folks here said I should pass on the mounting technique, so here it is.
Tip #1: BSE detection and Carbon I'm shooting with a Robinson BSE detector. It gives a more "real" illumination than SE and has the benefit of not "seeing" carbon. The specimen is normally mounted on top of a minuten (entomological) pin which is clamped in a 12mm "vice-stub" and tilted at 90 degrees in the SEM. On my Cambridge S120, the top of the stage mechanism is now out of view but you still see the bottom plate below. I've covered that with a piece of alfoil which has been painted with thinned-out carbon dag, making it a perfectly black, "studio-shot" background when using BSE.
Tip #2: No unsightly props Further, instead of mounting the specimen on the usual minuten pin, I've fixed it to the end of the lead from a clutch pencil (draughtsman's pencil), so the pin is also virtually invisible under BSE. Doesn't the pencil lead get coated too? I've taken a medium-bore tip from a syringe, held it in an alligator clip, and fed the pencil lead down into it leaving just the specimen exposed for coating.
Tip #3: Accuracy in mounting Mounting very small specimens on a pin-tip is difficult at the best of times. As the animation sequence is to be in stereo, the specimen must be at exactly the right angles to the pin and we don't want to see any excessive blobs of dag. You'll need a small bench vice, micrometer screw guage and double sided tape (DST). Lay your stereomicroscope down on its front with the head reversed so that the eyepieces point up and the light path is (roughly) horizontal. Clamp the side of the screw guage into the side of the vice and place it in front of the 'scope. Secure your specimen to a foam block with crossed minuten pins and secure the block in position to the opposing pole of the screw guage with DST. Fix the mounting pin to the mobile shaft of the screw guage with DST. Watching it all through the 'scope, do a dummy run to ensure that everything lines up. Screw the pin up, apply a meniscus of dag to the end of the pin and screw it down to the specimen. Leave to dry.
Tip #4: Centre of rotation Because this sequence includes rotating the animal 360 degrees, the pin has to be at the centre of rotation of the stage. However, in my 12mm "vice-stub", the vice is offset. Instead, I'm using a pop-rivet. Push the nail out and use the rivet and its collar; the rivet has the same diameter as the shaft on a normal 12mm grooved stub. Put the pin down the bore and dag it into a central position.
Hope you find these tips helpful and if you're in Oz around September, drop into the Australian Museum, Sydney, to see the stereo animation sequence of the spider (unfortunately, I'll be gone by then).
Geoff Avern Manager Microscopy Labs Australian Museum Sydney, Australia
Dear fellow microscopist I have had a suggestion from one of my colleagues about staining of nerve cells. He recommends including tannic acid in the fixative (paraformaldehyde/gluteraldehyde) at a concentration of 0.25% to 2% and then to use phosphotungstic acid to stain sections or use UA as a block stain prior to embedding. I have no idea how well this will work but apparantly tannic acid stains membranes clearly. I hope you have some success
Cheers Nikki Bock Dept. of materials engineering & materials design University of Nottingham Nottingham NG7 2RD Email: emznjb-at-emn1.nott.ac.uk
Can anyone advise on best technique for preparation of Chlorides (polished sections), as these materials are extremely soft and highly rippable resulting in abundant cavities in polished section. Several sample prep techniques have been tried without success.
John Yes it can be done To do this, put a drop of fixed virus suspension on to your coated grid for about 60 seconds. Than very slowly suck up virus suspension with a whatman filter paper.(cut filter paper into triangle, with sharp arrow point like tip.) Leave to dry for about 60 seconds.
Than float grid containing virus onto a drop of 0.05M sodium cacodylate buffer. (2 x 2 minutes wash)
Again wash with double distilled water. (2 x 2 minutes)
Negative stain for 30 seconds with PTA.
Dry and view.
All the best
Vijay H Bandu University of Natal Centre for Electron Microscopy Private Bag X01 Scottsvile 3209 South Africa
Hi Marty I seem to be havivig difficulty with your e-mail address. thought I might reach you here. I am trying to find a source for dowty window seals for our polaron CPD.You suggested All Seals in Santa Ana. Ca. I contacted them, but they require the Dowty part number or their own. Which Idon't have. Do you have that number? Any help would be greatly appreciated. Reply to ToogoodM-at-em.agr.ca
ToogoodM-at-em.agr.ca Agriculture and Agri Food Canada Kentville Research Centre Kentville N.S. B4N 1J5
Message-Id: {3.0.1.32.19970529084940.006d08e8-at-biotech.ufl.edu} X-Sender: sdw-at-biotech.ufl.edu X-Mailer: Windows Eudora Light Version 3.0.1 (32)
I have archived two discussions you may find useful. One is on thinsectioning bone, and the other undecalcified teeth. Go to the url listed at the bottom of this message and click on the "Tips & Tricks " link. Do a search for "bone" and there will be a number of files listed. The two most useful will be "teeth.html" and "thinbone.html". Good luck
At 05:18 PM 5/28/97 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 352-392-1295 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
I'm currently evaluating replacements for my Kevex EDS detector with a Be window. I use a Kevex 4505P Pulse Processor which dumps the signal down to a 4pi SC-2 multichannel analyzer for spectral analyses and imaging.
My goal is to either change my Be window to a thin film window or replace my current detector with one equipped with a thin film window for light element analyses.
Does anyone know if:
1) The Be window of the Kevex Model 2003 EDS detector is replaceable with a thin film window?
2) Can the Kevex 4505P Pulse Processor handle light element pulse throughput?
3) Are there EDS detectors available with thin film windows which are adaptable to the 4505P Pulse Processor?
Any ideas or references out there would be highly appreciated.
I hope we haven't missed the deadline for the American Lab article. Please find our submission attached. I will Federal Express the corresponding 35mm slides today. Sincerely, -Jennifer Robinson Product Manager, Fluorescence Microscopy
At 07:01 PM 5/28/97 -0400, Scott Walck wrote: } } The question that I have about charging in an SEM is this. If you set up the } imaging conditions for the sample not to charge at a high scan rate i.e. TV } rate, and then drop it to a slow scan (as you do in an analog machine when } recording an image), why does the image now charge?
As John Bozolla said, the beam is dwelling longer at each spot allowing more charge to build up. I think this would be related to the old V=I*R equation. In fast scanning, the peak current (electron dose) before the beam moves away and the sample has a chance to discharge is relatively small so the resultant voltage would be small. At slow scans (say 30-100 times slower) the voltage should be that much higher and would become noticeable. Warren E. Straszheim Co-Director of Ambience for BIBLE-at-VIRGINIA.EDU
He who has knowledge spares his words, and a man of calm understanding is of a calm spirit. - Pr 17:27 Even a fool is counted wise when he holds his peace; when he shuts his lips, he is considered perceptive. - Pr 17:28 It is better to keep silent and be thought a fool than to open your mouth and remove all doubt. - Murphy
You can also add Kheops from Noesis S.A. and Noesis Vision Inc. The product is now being demonstrated commercially, it will be available in June and on display at the Microscopy show in Cleveland in early August.
Regards,
At 08:32 PM 5/28/97 +0200, Heike Buecking wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
---------------------------------------------------------------------------- ------------- Luc Nocente Tel: 514 345 1400 Noesis Vision Inc. Fax: 514 345 1575 e-mail: ln-at-noesisvision.com http://www.noesisvision.com 6800 Cote de Liesse, Suite 200 St-Laurent, PQ H4T 2A7,Canada ---------------------------------------------------------------------------- -------------
Dear Reembedders, To successfully reembed thick sections (Araldite, Epon, LR White) one must understand the process. 1. Interface - The interface between the slide and the sections to be reembedded is of crucial importance. Sections will stick tenaciously to super clean glass slides, but will beautifully depart on command from the correct interface. The best interface is gelatin (no additives when subbing slides - may interfere with the LM stains). The interface cannot present a "soft" mixture of epoxies, perhaps created by improper polymerization due to alcohol, acetone, or propylene oxide left on the slide. 2. Not all epoxy mixtures are suitable for reembedding. Araldite, some epon replacements containing dilutents or plasticicers my remain too elastic, and the face of the section will deform when detaching. 3. Coefficient of thermal expansion - the most successful (ours is 99.9% successful, we do not loose sections) of detachment methods. Some people use cold, we use heat. We prefer heat, it is simple, and the glass becomes fluid, so to speak, preventing cold glass chips from becomeing part of the new block. Briefly, the rates of expansion between block and slide must vary widely. The slide is rapidly cooled or heated, the epon lags behind, and detachment occurs. Detaching hot blocks from hot slides is not as successful. Frequently the sections deform (bad for correlative microscopy). 4. The water tower effect - Capsules which are reversed over sections on slides (capsules are filled with epoxy) must be of a certain size. Skinny, small capsules, result in a lot more seeping epoxy around the edges, than fat short ones (the standard size of embedding capsules). Seeping epoxy causes "collaring" which makes detachment difficult. 5. Correlative microscopy of immunocytochemical materials (DAB) requires flat sections- Sections will move and warp while being remembedded making exact correlations difficult. This is best prevented by using a glass microscope slide (subbed) as a bottom slide on which the sections rests. Thick sections are dried down on this slide, stained and photomicrographed. The capsule is then inverted over the section. The oven temp should not go over 60degC. If sections are free-floating, as in a DAB reaction, they are placed on subbed microscopy slides, carefully wiped clear of excess epoxy, and covered with another slide which has been treated with mold release (Trennmittel from EMS). (I own no stock in EMS). The two slides are than weighted or clamped together, and polymerized. The top slide comes off easily, the sections is flat for photomicrography, and then reembedded with a gelatin capsule full of epon. I have run numerous trials on epon mixtures and found the Luft's medium mixture to be ideal. It is best to allow the epon mixture to rest for 30 min after acceleration, because it inhibits seeping and "collaring" around the section. 6. Original processing of tissue - If the original tissue is poorly infiltrated or suboptimally prepared, it is essentially "over". During the reembedding process, the new epoxy will bind to the old in an unpredictable manner and the section may warp badly, split when pulled off, etc. Many of the above statements have their origins in the writing of Lee and Neville - Handbook of Epoxy Resins, or in the publications of Causton and the people who hold patents for LR White, etc. If anyone wants a step by step method for reembedding of sections from a Vibratome or an ultramicrotome, please request. This is lots of fun, and lots of challenge. Reembedding could be a masochist's pradise! Actually TEM could be a masochist's best bet! Right? Bye, Hildy
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear all,
Wanting to control the presence of esterified pectins in plant cells I've tried the hydroxyalmine/ferric chloride reaction as described by Reeve (1959). I suppose this reaction is routinely used by many of us but.... I have a precipitation problem when I add to the fresh hydroxylamine reagent - i.e. a solution of equal volumes of sodium hydroxide (14g in 100ml EtOH 60%) and hydroxylamine hydrochloride (14g in 100ml EtOH 60%) - concentrated HCl in EtOH in order to acidify the reaction mixture. The precipitate I obtain is of cristalline structure and rather sticky... If you met this problem to when following exactly Reeve's protocole, if you were lucky in solving this problem or if you were not, etc... Please help ! Thanks Pascal
************************************ Pascal VEYS Laboratory of Plant Biology Catholic University of Louvain Place Croix du Sud 5 (bte 14) B 1348 Louvain-la-Neuve Belgium Phone : 0032 10473004 Fax : 0032 10473471 Email : Veys-at-bota.ucl.ac.be ************************************
************************************ Pascal VEYS Laboratory of Plant Biology Catholic University of Louvain Place Croix du Sud 5 (bte 14) B 1348 Louvain-la-Neuve Belgium Phone : 0032 10473004 Fax : 0032 10473471 Email : Veys-at-bota.ucl.ac.be ************************************
Sounds like embedding in resin, followed by diamond (or glass) knife sectioning might be worth a try. This technique has been used for decades as a means of preparing soft metals for metallographic examination. Reaction of the chlorides with the standard embedding resins might be an issue. Also, some resins need modest heating (~60 C) for curing.
Tom Malis Group Leader - Characterization Materials Technology Laboratory Natural Resources Canada Ottawa, Canada K1A 0G1
______________________________ Reply Separator _________________________________ Subject: SAMPLE PREP. OR MICROSCOPY Author: petra-at-iafrica.com at internet Date: 5/29/97 11:57 AM
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Can anyone advise on best technique for preparation of Chlorides (polished sections), as these materials are extremely soft and highly rippable resulting in abundant cavities in polished section. Several sample prep techniques have been tried without success.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Technician Certification 5/29/97 12:45 PM
In light of the recent inquiries to accreditation, I would like to let listserver members know there is a Certification Program for biological electron microscopy technicians available through the Microscopy Society of America. It involves a practical and written examination. The practical requires the technician to prepare and embed samples for resin, section and take pictures. The written exam is a comprehensive 100 questions pertaining to fixation, embedding, microscopy equipment and sectioning techniques. If you would like more information you may contact the MSA Business Office at 4 Barlows Landing, Pocassett, MA 02559, telephone 1-800-538-3672, fax 1-508-563-1211 or email at BusinessOffice-at-MSA.Microscopy.Com
Linda Chicoine Center for Cell Imaging Yale University 333 Cedar Street, PO Box 208002 New Haven, CT 06520-8002 phone 203-785-3646 fax 203-785-7226
I have not prepared chlorides alone, but have prepared (polished) metallic mounts with chloride inclusions. Typical metallographic techniques were employed except decane (high purity kerosene) was used instead of water. The first prep using water disolved the inclusions. Using non-polar lubricants and cleaning agents solved our problem.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Can anyone advise on best technique for preparation of Chlorides (polished sections), as these materials are extremely soft and highly rippable resulting in abundant cavities in polished section. Several sample prep techniques have been tried without success.
I have not prepared chlorides alone, but have prepared (polished) metallic mounts with chloride inclusions. Typical metallographic techniques were employed except decane (high purity kerosene) was used instead of water. The first prep using water disolved the inclusions. Using non-polar lubricants and cleaning agents solved our problem.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Can anyone advise on best technique for preparation of Chlorides (polished sections), as these materials are extremely soft and highly rippable resulting in abundant cavities in polished section. Several sample prep techniques have been tried without success.
Hi: Does anyone have any information on the Life Cell Corp which produces the CF-100 slam freezer. I need their phone #. Also has anyone used one or a similar model. Pros and cons would be appreciated.
Thanks, Charlie Murphy cmurphy-at-ggpl.arsusda.gov
There is a regular full-time position available in the Department of Biological Imaging (a Shared Scientific Service) at the Jackson Laboratory in Bar Harbor, ME.
Duties include operation and training of end users on a PC-based image analysis system and the performance of customer-directed analysis of biological research specimens. Routine maintenance and use of a confocal microscopy system will be a major requirement for this position. Also included is the regular maintenance and alignment of upright, inverted and stereo research-level microscopes, with the following optics: brightfield, darkfield, phase contrast, differential inteference contrast and fluorescence. A successful candidate would have a Master's degree (or equivalent experience and qualifications) in biological sciences and a minimum two years experience in confocal microscopy and/or microscopic imaging, experience in both being most desired. Experience in rudimentary computer programming is also preferred. Experience in electron microscopy and/or histology would be an asset. The applicant must be able to work independently in a multi-user facility and deal with people on a one-to-one basis. Individual will be expected to attend seminars and participate in interest groups disseminating information about current microscopic techniques.
Interested applicants may forward resumes to : Joanne Bradt
At 09:18 29/05/1997 -0400, you wrote: } I hope we haven't missed the deadline for the American Lab article. Please } find our submission attached. I will Federal Express the corresponding } 35mm slides today. } Sincerely, } -Jennifer Robinson } Product Manager, Fluorescence Microscopy } } (See attached file: Cell_des.doc) } } Attachment Converted: "C:\EUDORA\Attach\Cell_des.doc" } Even if English is not my mother language, I think this Email is a commercial advertisement. Specially when it's loading on my attach directory some files I must delete by manually acting. We are a lot of commercial company using this mailing list and we must respect the ame the list. Meme si l'anglais n'est pas ma langue maternelle, je pense que cet Email est a vocation commercial. Je n'admet pas de recevoir en attach un fichier que je devrai detruire en recherchant dans mes fichiers. Nous sommes plusieurs societes commerciales a utiliser cette facilite et nous devons respecter le fond de la liste. Best regards. --=====================_864938563==_ Content-Type: application/msword; name="Cell_des.doc"; x-mac-type="42494E41"; x-mac-creator="4D535744" Content-Transfer-Encoding: base64 Content-Disposition: attachment; filename="Cell_des.doc"
I hope your journey back to Germany was uneventful. I am glad we had a chance to discuss some of issues that create challenges for all of us.
One major point that we failed to discuss was the status of UL listing for the DM LB 30 and DM LB 100. This is a Key success factor for the DM LB. These instruments are effectively being kept out of hospital labs and research labs in hospitals because they don't have UL. This a problem in the 3 largest marked in the US. Los Angeles, Ca --- Chicago, IL --- and New York City. What is our progress and when can we expect to see these marks on the instrument? By the way, it would be nice for the LM, LSP and LP as well.
I understand that another vendor is being considered to supply us with an HBO power supply. Let me add that this power supply should also be UL listed. CSA is also acceptable, but the UL is preferred in the States. Of course, just the opposite occurs in Canada.
Jurgen, I've recently been working with AgBr(x)Cl(1-x). What a pig of a material! The best way of making sections is to use a microtome; polishing using diamond impregnated plastic, finishing with syton, works reasonably well if you can avoid embedding any particles in it. Other problems, apart from the softness, is the reactivity (reacts with virtually everything; Au, Ag and Pt are the only 'safe' metals which can make contact with it), and decomposition of the material when hit with light, electron beams (you can watch the composition change using EDX), and ions (so any ion milling is out). Put this together with an incredibly high coefficient of thermal expansion and you have the material scientist's material from hell. Good luck!
Richard Beanland, GMMT Caswell, Towcester, Northants NN12 8EQ UK
} } } Can anyone advise on best technique for preparation of Chlorides (polished } sections), as these materials are extremely soft and highly rippable resulting } in abundant cavities in polished section. Several sample prep techniques have } been tried without success. } } Regards, } Jurgen Paetz } 9 shell Rd. } Bloubergrand 7441 } CAPE TOWN - R.S.AFRICA } } REPLY: Jurgen Paetz } E-Mail: {petra-at-iafrica.com}
Scott D. Walck wrote: } The question that I have about charging in an SEM is this. If you } set up the imaging conditions for the sample not to charge at a high } scan rate i.e. TV rate, and then drop it to a slow scan (as you do } in an analog machine when recording an image), why does the image } now charge?
John J. Bozzola answered: } Because in a slower scan mode the beam dwells longer on the specimen } permitting more of a static charge to build up; the charge collects } with time.
The observations reported in "Observation of voltage contrast at grain boundaries in YSZ" by Charlotte Clausen (now C. C. Appel) and me in Micron and Microscopica Acta vol. 23 (1992) p. 157-158 illustrates the point made by John. We show a sample of YSZ in which you can see the grain boundaries imaged with a dark contrast when the beam current is 6 nA and the SEM is operated at slow scan rate (50 s per frame). The grain boundaries obtain the dark contrast because they conduct the charge away easier than the grain interior and therefore obtain a positive potential with respect to the grain interior.
The contrast disappears if either the beam current is lowered at constant scan rate OR the scan rate is increased at constant beam current.
-at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- J. B. Bilde-Soerensen Materials Research Department Risoe National Laboratory DK-4000 Roskilde Denmark
I apologize for posting what was a commercial press release. I was responding to a request for submissions from Barbara Foster and I inadvertantly posting to the entire list instead of only Barbara. Please be assured that it was unintentional and I have the utmost respect for the aim of the list. -Jen R., Scanalytics
I need a lens with a "C"-mount for a Sony Model XC-77 analog video camera. Can anyone recommend a source? A macro lens would probably work the best for my application (image analysis).
Thank you.
*****************************************
Delilah Wood United States Department of Agriculture Western Regional Research Center 800 Buchanan Street Albany, CA 94710
Why is it that when I do double immunostaining with two successive monoclonals I get inconsistant results with 'stains' that are fine by themselves? If I do a polyclonal followed by a monoclonal, no problems. What is the story? Are there ways around this? Thanks in advance!
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (908)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
Delilah Wood wrote: } } I need a lens with a "C"-mount for a Sony Model XC-77 analog video camera. } Can anyone recommend a source? A macro lens would probably work the best } for my application (image analysis). } } Thank you. } } ***************************************** } } Delilah Wood } United States Department of Agriculture } Western Regional Research Center } 800 Buchanan Street } Albany, CA 94710 } } Tel: 510.559.5653 } Fax: 510.559.5777 } Email: wood-at-pw.usda.govDear Delilah,
We found a wide variety of c-mounts available from Diagnostic Instruments in Sterling Heights, MI. Phone number: (810)731-6000. Their people were also very knowledgeable about different types of applications and which lens was needed for which situation. They also have an intriguing zoom system.
Good luck, Barbara Foster Microscopy/Marketing & Education
Caveat: We are not a vendor or promotor for Diagnostic Instruments; only an interested user.
You can contact LifeCell Corporation at: Tel. (713) 367-5368 Fax (713) 363-3360
I used to work at LifeCell, and helped develop the CF100 and the molecular distillation drying device, so I'm probably not the one to give you an unbiased opinion. I'm sure LifeCell can give you some references to call.
I *will* take this opportunity to say that cryofixation has never been easier or more reproducible than with the CF100. This is because of the microporcessor control, which monitors all of the parameters of cryofixation and permits fixation only after all of the conditions have been satisfied. Also, the microprocessor automatically regenerates the cryofixation surface after it's been used by heating it under a vacuum and then re-cooling it to prepare for another fixation run. The throughput on the CF100 is truly amazing -- as I recall, you can do a cryofixation about once every 2-1/2 to 3 minutes.
Best of luck to you. If I can be of any further assistance, don't hesitate to contact me off-list at RCHIOVETTI-at-aol.com.
The Pathology Department here at the Boston Univ School of Medicine has asked me to post a notice that their Hitachi 11C TEM is freely available. It has been under service contract for most of its life although not during the last five years. Interested folks should respond at busmpath-at-bu.edu
Dr. Steven Barlow EM Facility/Biology Department 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/EM_Facility
Are you sure it is one micron gold particle? It does not sound right .. It is almot three times the size of an average cell or 50x the size of viral particle.. I believe it should be one nano meter.. That I may be able to help you with...
At 02:08 PM 5/28/97 -0400, Sharon Godkin wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I did send email to ListServer-at-... for unsubscribe by using the required command several times. But I am still on the list. Could you remove me from the list?
Are you working at the light level or the EM level?
If you are working with immunoEM, it sounds like the problem with your double immunolabeling may be cross-reactivity between the monoclonals and whatever you're using for the subsequent steps. This is frequently the situation with protocols that use, for example, mouse monoclonal primary antibodies, then goat anti-mouse Ig's followed by some method to localize with colloidal gold (maybe biotinylated goat anti-mouse, followed by streptavidin-colloidal gold?).
If you *are* working at the EM level, you might want to consider doing a double-sided labeling. In a nutshell, place the sections on uncoated hexagonal mesh nickel grids. Do one labeling run on one side, going through all the way to distilled water rinsing and drying. Flip the grid over, then perform the second labeling on the other side, using gold of a different size. This "compartmentalizes" each label, and greatly reduces the cross-reactivity.
Hope this helps. Feel free to contact me off-list if you need additional details.
Best regards,
Robert (Bob) Chiovetti, Ph.D. RCHIOVETTI-at-aol.com
} any suggestions where I can buy a water trap for my liquid carbon dioxide } tank attached to my CPD? } At one time Tousimis (spelling?) sold an in-line unit. I saw the set up at the EM facility of University of Wisconsin -at- Madison.
} ______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 The College of New Jersey fax: (609) 771-2674 Trenton, NJ 08650-4700
The Microscopy & Microanalysis '97 Search Engine is now on-line at
http://www.msa.microscopy.com
using this engine you may search the M&M' 97 program by Author, Title Keywords, Symposium Name or Day of the Week and find out when and where each presentation will occur.
Contrary to last year, the abstracts for all papers for the meeting will not be on line this year as the publication and organization of that aspect of the meeting is being coordinated by our new Journal.
A committee within the Dutch Microscopy Society (NVvM) is working on the general issue of calibration of microscopes (SEM, TEM, LM, AFM, STM). Special attention is paid to calibration in relation with accreditation. We would like to contact a microscopy lab that is certified or users of certified microscopes in order to exchange some ideas. Any general information on the issue is welcomed as well.
Thanks in advance.
Bart Nelissen Akzo Nobel Central Research Dept. Applied Physics Microscopy eMail: Bart.B.J.Nelissen-at-akzo.nl
I think it should work fine if both monoclonals are directly conjugated. But if you are using conjugated secondaries you will get your second primary sticking to your first secondary. You will also get your second secondary sticking to your first primary. So you will get various states of crossover. I think after you put on your first primary and secondary, block with 10x excess of anti-mouse Fab fragments to bind up all the free sites should work.
Bob Underwood Morphology Core U of W Seattle, WA, USA
On Fri, 30 May 1997, Geoff McAuliffe wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear Microscopists: } } Why is it that when I do double immunostaining with two } successive monoclonals I get inconsistant results with 'stains' that are } fine by themselves? If I do a polyclonal followed by a monoclonal, no } problems. What is the story? Are there ways around this? } Thanks in advance! } } Geoff } -- } *************************************************************** } Geoff McAuliffe, Ph.D. } Neuroscience and Cell Biology } Robert Wood Johnson Medical School } 675 Hoes Lane Piscataway, NJ 08854 } voice: (908)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu } *************************************************************** }
Steve, In the past I have purchased water filters and particle filters for CPD from Tousimis Res. Corp. 1-800-638-9558 is the last ph.# I have for them. (No affiliation, of course) cheers ed
Edward J. Basgall, PhD The Pennsylvania State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618
} any suggestions where I can buy a water trap for my liquid carbon dioxide } tank attached to my CPD? } } TIA } } steve } } --------------------------------------------------------------------- } } Dr. Steven Barlow } EM Facility/Biology Department } 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/EM_Facility
Please can you advertise the following posts which will come into effect from October 1997.
Thanks, Rik Brydson
************************************************************** School of Process, Environmental and Materials Engineering, University of Leeds, U.K.
Research Opportunities in Materials Characterization
Two positions are currently available:
o Earmarked EPSRC Ph.D. Studentship in Materials Characterization - Analytical Electron Microscopy and/or Surface Analysis applied to a wide range of materials (carbons/ ceramics/ electroceramics/ catalysts/ non-ferrous alloys and steels). Candidates should possess a 2.1 degree or higher in either Materials, Physics, Chemistry or related discipline.
o 3 Year EPSRC funded Postdoctoral Position on the modelling of electron energy loss near-edge fine structure (unoccupied electronic structure). Candidates should possess a Ph.D. in the physical sciences or an engineering discipline and have experience in two or more of the following: solid state physics/chemistry, electron microscopy and/or computing/programming.
For further details concerning both posts please contact: Dr Rik Brydson, Electron Optical Unit, Materials, Leeds LS2 9JT, U.K. (email: mtlrmdb-at-leeds.ac.uk/ tel: 0113 233 2369) ************************************************************** _____________________________ Dr. Rik Brydson, University Research Fellow, Electron Optical Unit, Department of Materials, School of Process, Environmental and Materials Engineering University of Leeds, Leeds LS2 9JT, U.K.
I am looking for an electrolyte in order to prepare some TEM thin foils of Al-Mg alloys. I have the following constraints for the eloctrolytic polishing solution : -polishing at room temperature. -solution that does not attack copper.
In addition what is the influence of the metal of the cathod (Platinum, stainless steel, copper, ...) on the result of the electrolytic polishing?
Frederic Basson McMaster University Department of Materials Science and Engineering 1280 Main Street West Hamilton, Ontario L8S 4L7, Canada Tel : (905)-525-9140 Ext 24862 Fax : (905)-528-9295 Email : bassonf-at-mcmail.CIS.McMaster.CA
We are using Tousimis otl/water filter now. The information on filter is:
Replacement Element for oil/water filter Cat. #8782A 2211 Lewis Ave. Rockville MD 20851 Tel: #301-881-2450
Wang
On Sat, 31 May 1997, Donald Lovett wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } On Fri, 30 May 1997, Steve Barlow wrote: } } } any suggestions where I can buy a water trap for my liquid carbon dioxide } } tank attached to my CPD? } } } At one time Tousimis (spelling?) sold an in-line unit. I saw the set up } at the EM facility of University of Wisconsin -at- Madison. } } } } ______________________________________________________________________ } Donald L. Lovett e-mail: lovett-at-tcnj.edu } Assoc. Professor, Dept. of Biology voice: (609) 771-2876 } The College of New Jersey fax: (609) 771-2674 } Trenton, NJ 08650-4700 } } } }
We are using Tousimis otl/water filter now. The information on filter is:
Replacement Element for oil/water filter Cat. #8782A 2211 Lewis Ave. Rockville MD 20851 Tel: #301-881-2450
Wang
On Sat, 31 May 1997, Donald Lovett wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } On Fri, 30 May 1997, Steve Barlow wrote: } } } any suggestions where I can buy a water trap for my liquid carbon dioxide } } tank attached to my CPD? } } } At one time Tousimis (spelling?) sold an in-line unit. I saw the set up } at the EM facility of University of Wisconsin -at- Madison. } } } } ______________________________________________________________________ } Donald L. Lovett e-mail: lovett-at-tcnj.edu } Assoc. Professor, Dept. of Biology voice: (609) 771-2876 } The College of New Jersey fax: (609) 771-2674 } Trenton, NJ 08650-4700 } } } }
I am experiencing an imaging problem with Photoshop 3.0.
Images are acquired by scanning in TEM negs using a Polaroid Sprintscan 45 and a Power Mac 8500. Files are saved on the Mac hard drive in TIF format for IBM's. No problem opening the IBM files from the Mac hard drive, but when the files are copied onto an IBM-formatted ZIP disk, they open as segmented images on the Mac -- like someone cut the images into strips and jumbled up the order.
I can open the images from the IBM ZIP disk using NIH Image and PageMaker on the Mac but NOT Photoshop. Now, if I drag the files from the IBM ZIP disk back onto the Mac hard drive, they open properly. I also noticed this problem on IBM formatted JAZ drive as well.
What's going on and how can this be remedied? Is this an Iomega glitch?
Thanks,
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 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 ATTEMPTED TO DO RESOLUTION MEASUREMENTS USING A LAB6 FILAMENT. I WAS ABLE TO RESOLVE A FEW MOCRONS USING A TUNGSTEN FILAMENT AT VARIOUS CURRENTS, HOWEVER WITH THE LAB6 FILAMENT I HAVE HAD PROBLEMS FOCUSING THE MICROSCOPE AND HAVE NOT BEEN ABLE TO RESOLVE THE MICRON STRUCTURES ON THE TEST SAMPLE. DOES ANYONE HAVE A REASONABLE EXPLANATION ? THE SEM IS A ISI SX40.
I am in charge of our EM facilities and have been asked to compare our charge schedule with other lab's. We have a Philips 201 and a 420,as well as an SEM. We do all types of specimen preparation (including freeze etching and cryosectioning) and dark room work. Thanks for any info which can be directed to me at the adress above or to revelj-at-cco.caltech.edu JP Revel.
} I ATTEMPTED TO DO RESOLUTION MEASUREMENTS USING A LAB6 FILAMENT. I WAS } ABLE TO RESOLVE A FEW MOCRONS USING A TUNGSTEN FILAMENT AT VARIOUS } CURRENTS, HOWEVER WITH THE LAB6 FILAMENT I HAVE HAD PROBLEMS FOCUSING } THE MICROSCOPE AND HAVE NOT BEEN ABLE TO RESOLVE THE MICRON STRUCTURES } ON THE TEST SAMPLE. DOES ANYONE HAVE A REASONABLE EXPLANATION ? THE SEM } IS A ISI SX40.
If you do not have good enough vacuum, chromatic aberration will effectively enlarge the spot size and destroy your resolution.
Also, the gun circuitry may need to be modified to permit you to effect good emission currents. Was your filament installed by service personnel and the scope properly set up to accept the filament?
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
I expect that the resolution change has nothing to do with that change-over, rather it points to a contamination/ stigmatism problem. Also, because of the greater brightness from the LaB6 you would probably use the condensor more focused and the smaller probe size should actually improve resolution. It is assumed that other vital working parameters (distance, kv, beam current, specimen type) are unchanged. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 350+ Links, MSDS ************************ http://www.proscitech.com.au ---------- } From: Bruce Brinson {brinson-at-rice.edu}
} Subject: EM LAB6 CONVERT. } Date: Tuesday, 3 June 1997 6:08 } I ATTEMPTED TO DO RESOLUTION MEASUREMENTS USING A LAB6 FILAMENT. I WAS } ABLE TO RESOLVE A FEW MOCRONS USING A TUNGSTEN FILAMENT AT VARIOUS } CURRENTS, HOWEVER WITH THE LAB6 FILAMENT I HAVE HAD PROBLEMS FOCUSING } THE MICROSCOPE AND HAVE NOT BEEN ABLE TO RESOLVE THE MICRON STRUCTURES } ON THE TEST SAMPLE. DOES ANYONE HAVE A REASONABLE EXPLANATION ? THE SEM } IS A ISI SX40. } } THANK YOU
Email: cesa-at-compass.com.ph Name: Christian Eric S. Abaya
School: University of the Philippines
Question: I am a graduate student of the University of the Philippines, and I } am now starting doing my masteral thesis which is somehow related on } electron microscopy of microorganisms (local isolates-bacteria, fungi and } actinomycetes). I am experiencing some problem isolating and preparing a } pure strain for } TEM and SEM observation. Can you provide me some basic technique in } isolating and processing microorganism (for TEM and SEM). } Your immediate response will be very much appriciated. Thank you } very much.
From experience I could say following - as in serviced by me Philips microscopes it is possible nicely to visualize the so called beam crossover which is showing image of the filament spot after Wehnelt - I observed following: - the Lab6 is more sensitive to Wehnelt - tip distance adjustment and tip centering as W. - close-to-saturation-point looks like malthese cross with 4 bright arms not like the torus by W-filament - if the Wehnelt distance is not good you can see on image status like saturation (no more light, by more heating) but you are on one of the arms or in the middle but arms are still bright - this causes the image to be not sharp - like the shadows of soccer players on the night-illuminated stadion. - if centering is wrong - you can have by saturation even to spots (one from arm one from centertip) - by not-centered W-filament you have banana shape which anyhow can reach spot-saturation. - also if you just exchanged filament - maybe you can not reach saturation value of the heating current ? - did this microscope ever worked with Lab6 ? kind regards Krzysztof M. Herman PHILIPS E.O. Service Poland LabSoft S.c. 05-500 Piaseczno, 21 Kosciuszki Str. tel/fx: (48 22) 7502024, 7502028, 7570671 fax only: (48 22) 483787, Email: labsoft-at-ikp.atm.com.pl
} Has anyone a suggestion on how to prepare a metal sample to get a strong } electron channeling effect?
I presume you're intending to look at the specimen on an SEM? I'd think that something with nice, large crystals would make a good specimen - grind and polish a flat surface for the channeling. Perhaps a brass would work?
} I ATTEMPTED TO DO RESOLUTION MEASUREMENTS USING A LAB6 FILAMENT. I WAS } ABLE TO RESOLVE A FEW MOCRONS USING A TUNGSTEN FILAMENT AT VARIOUS } CURRENTS, HOWEVER WITH THE LAB6 FILAMENT I HAVE HAD PROBLEMS FOCUSING } THE MICROSCOPE AND HAVE NOT BEEN ABLE TO RESOLVE THE MICRON STRUCTURES } ON THE TEST SAMPLE. DOES ANYONE HAVE A REASONABLE EXPLANATION ? THE SEM } IS A ISI SX40. } } THANK YOU
If the filament isn't set up correctly in the gun, it will generate a large fuzzy spot. It may be that you just need to increase the filament current to get proper saturation - but be careful not to blow the filament. This assumes that the gun circuitry is capable of driving LaB6 and that you have a good vacuum.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have been asked by Dr. John E. Johnson, Jr., Editor-in-Chief of Microscopy Research and Technique (MRT), to solicit selected articles addressing recent advances in our understanding of Silver Halide Photographic Processes. The invited articles would be the basis for a topical issue of MRT entitled "Microscopic Research of Silver Halides and Related Dispersed Systems", on which I would serve as guest editor. Research in this field may give a number of excellent examples of successful solution of highly diverse and complicated tasks of imaging science of more than 150 years history and modern high technology focused to the next century. The term "microscopy techniques" may cover all known methods of light microscopy, electron microscopy (stationary and scanning methods), scanning probe techniques, image analysis and so on.
In this regard, I am writing to invite you, either alone or in conjunction with a collaborator(s) of your choice, to submit articles discussing some of the following topics concerning methodology, sample preparation and applications of microscopy in studies of silver halides and related dispersed systems (small particles and clusters of metals and metal sulfides, polymer gels and latexes, dyes, etc.):
1. Microscopic insight to mechanisms of nucleation and growth of silver halide crystals of photographic emulsions and model systems. 2. Crystalline and defect structures and phase composition of silver halides: impact of microscopy. 3. Structural and analytical characterization of silver halides and related dispersed systems by imaging and spectroscopic microscopy techniques. 4. Microscopy and ultramicroscopy studies of mechanisms of chemical and spectral sensitization and latent image formation. 5. Development and processing of silver halide photographic systems as studied by microscopy methods.
A manuscript length limitation is set at about 30 double-spaced typewritten pages, plus micrographs. An Abstract should be included. Each manuscript will be refereed, and therefore will be suitable for inclusion in Grant Proposals or Renewals. The MRT Instructions to Contributors (enclosed) should be used as a guide when preparing the manuscript. Note that the journal has a format of 8 1/4" by 11", and the figure size is 6 3/4" wide by a maximum of 9" high for a full page width figure, or 3 5/16" wide by a maximum of 9" high for a single column width figure. Therefore, your figures should be trimmed to these dimensions if you wish them to be reproduced at the same size as the originals. There is a charge to authors for color figures, but no charge for black and white figures, regardless of the number. You would need to include copyright release forms signed by the publisher for any previously published figures. Please examine MRT for examples of what we are looking for in topical papers. If you would be willing to participate in this venture, I would appreciate knowing your intentions by 1.07.97. I would need your Article Outline (with approximate number of pages and figures) by 15.09.97., and I would, at that time, make available to you a list of other contributors and article titles. The completed manuscript would be due in my office by 1.11.97. All manuscripts are peer reviewed and may require revision. The topical issue will be published within 4-6 months (this is part of our agreement) after the manuscripts are received at the publisher's office in New York (John Wiley). I hope you will agree to contribute to what we believe will be a fascinating and beneficial update of current knowledge in Silver Halide Photographic Processes.
Sincerely,
Vladimir Oleshko ********************************************************** V.P. Oleshko, Ph. D. e-mail:oleshko-at-uia.ua.ac.be Micro-and Trace Analysis Centre Tel.:+32-3-820.23.64 Chemistry Department FAX :+32-3-820.23.76 University of Antwerp (UIA) Universiteitsplein 1 Antwerpen-Wilrijk B-2610 Belgium ***********************************************************
John, Do I understand you to say that when you read from the ZIP into MAC photoshop the image comes up in jumbled strips, but when you drag the image from the ZIP to the hard drive then the image reads in correctly? If that be the case, I would wonder if there is something wrong in the ZIP drivers or hardware.
On the other hand, if you only see the problem reading into PC Photoshop from the ZIP, then I would wonder if the problem is elsewhere.
Some years ago, I did a TIF converter for our KEVEX images. I had a dickens of a time trying to please all of the TIF readers out there. There were so many options available for TIF files and the readers did not handle all of the tags correctly. I checked PC Word, PageMaker and a few other readers and got various results. Some were much easier to please than others. And when I had them 'happy', some MAC applications had problems with my format.
Since then, TIF writers and readers have undoubtedly improved so that they support more of the standard correctly, but there still could be problems depending on the version.
Off hand, I wonder if your images are being stored in strips (as is often the case) and that the strip addresses are getting jumbled. Sometimes there are advanced options for saving the images that allow you to specify the strip size, e.g., 8K or 32K; I wonder if that would have an impact on what you see. Also, are you able to save it to any other image formats or to turn off the strip option? I don't think that either GIF or Windows BMP formats use strips.
If you still have problems, you might try sending an attached image to me (not to the list) or tell me where I can access one and I can take a look at the internals and see what I can see.
At 01:51 PM 6/2/97 -0600, you wrote: } } Calling all imaging gurus, } } I am experiencing an imaging problem with Photoshop 3.0. } } Images are acquired by scanning in TEM negs using a Polaroid Sprintscan 45 } and a Power Mac 8500. Files are saved on the Mac hard drive in TIF format } for IBM's. No problem opening the IBM files from the Mac hard drive, but } when the files are copied onto an IBM-formatted ZIP disk, they open as } segmented images on the Mac -- like someone cut the images into strips and } jumbled up the order. } } I can open the images from the IBM ZIP disk using NIH Image and PageMaker } on the Mac but NOT Photoshop. Now, if I drag the files from the IBM ZIP } disk back onto the Mac hard drive, they open properly. I also noticed } this problem on IBM formatted JAZ drive as well. } } What's going on and how can this be remedied? Is this an Iomega glitch? } } Thanks, } #################################################################### } John J. Bozzola, Ph.D., Director ---------------------------------------------------- Warren E. Straszheim 270 Metals Development, Ames Lab/ISU, Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-3091
We have a refurbished jsm 35C for sale at the present time for $13,750. If interested please give us a call at 770-448-3200. We need to make room for other instruments now. Talk to Mark Rigler when you call. Thanks
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Has anyone a suggestion on how to prepare a metal sample to get a strong } electron channeling effect?
Usually an electropolished sample gives a strong electron channeling contrast I know this works for steels, iron, Al, CuAg alloy and many others.
}
Dr. Ijaz A. Rauf Department of Physics, University of Alberta, Edmonton, Alberta, Canada, T6G 2J1.Ph:(403)492-3041 Fax:(403)492-0714 E-mail:irauf-at-phys.ualberta.ca **** Love For All ** Hatred For None **** I Express My Opinions Only *** ************************************************************************ * Ahmadiyyat, in fact, is the true Islam revealed to Muhammad (PBUH) * * You can watch for yourself on satellite TV, for details about time * * and channels in your region visit URL http://alislam.org/mta/ * ************************************************************************
} Klaus } =20 } The Biomaterials Science Group } Department of Oral and Dental Science=20 } in co-operation with the Physics Department } University of Bristol } Lower Maudlin Street, Bristol, BS1 2LY, England } =20 } =20 } =20 } For the project } =20 } "Protein-Biomaterials-Interfaces Investigated with Atomic=20 } Force Microscopy" } =20 } we are looking for } =20 } a PhD Student } =20 } with a background in Physics or Materials Science or Chemistry or Biolo= gy } or Engineering } =20 } or a student of related areas. } =20 } We expect: } =B7 EU citizenship (non-EU citizens can be considered if they } provide funding which covers the difference between the=20 } oversees fee and the home fee) } =B7 Upper Second Class degree or better } =B7 Enthusiasm for the work in the area of biomaterials interfaces } =B7 Eagerness to explore new areas, initiative and result oriented work } =B7 Starting date 1st October 1997 } =B7 Open, cooperative character for the work in an international team } =20 } We offer: } =B7 Research at one of the leading British research universities in att= ractive surroundings } =B7 Top departments } =B7 PhD fees are paid } =B7 Maintenance and travel grant (co-operation with a university in Cal= ifornia possible) } =B7 Applied scientific research on a high level } =B7 Intensive supervision } =B7 Work in international teams with an excellent working atmosphere } =B7 Future oriented research area } =B7 Premium (=A3 2000) from industrial sponsor paid upon successful com= pletion of PhD research programme } =20 } Closing date for applications 15th June 1997 } =20 } Applications should be submitted to: } =20 } Dr. Klaus D. Jandt =09 } Senior Lecturer } Dental Materials Science and Biomaterials } University of Bristol =09 } Department of Oral and Dental Science =09 } Lower Maudlin Street =09 } Bristol, BS1 2LY, UK =09 } Phone: ++ 44 (0)117 9 28 44 18 } Internet: K.Jandt-at-bris.ac.uk } =20 } ----------------------------------------- } Dr. Klaus D. Jandt } Senior Lecturer } Dental Materials Science and Biomaterials } University of Bristol } Department of Oral and Dental Science } Lower Maudlin Street } Bristol, BS1 2LY, UK } Phone: ++ 44 (0117) 9 28 44 18 } Fax: ++ 44 (0117) 9 28 47 80 } Internet: K.Jandt-at-bris.ac.uk } WWW: http://mail.bris.ac.uk/~omkdj/ } =20 } =20
On 06/02/97 19:12:06 you wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Our Safety officer is concerned about us decanting the liquid nitrogen from our dewar into our coldstage using a styrofoam cup. Does anyone have any suggestions on where we might purchase a cryo ladle of some sort. The cryo gloves that we have, are too cumbersome when using such a small styrofoam cup for the small quantity that we decant at a time.
Any suggestions would be greatly appreciated.
Susan
Susan Carbyn Atlantic Food and Horticulture Research Centre Agriculture and Agri-Food Canada Kentville, Nova Scotia B4N 1J5 Canada
A colleague would appreciate input on chemical etching Ge(111). He has used HF10%+HNO3 90% for Ge(001) and obtained nice thin regions around the hole, but for Ge(111) got only pinholes with this solution even when cutting the strength several times.
Any suggestions?
W. Sinkler
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Wharton Sinkler PhD Department of Materials Science and Engineering Northwestern University 2225 North Campus Drive Evanston, IL 60208-3108 tel: (847) 491-7809 fax: (847) 491-7820 email: sinkler-at-apollo.numis.nwu.edu
I've finished my senior research project and have an abstract for the MSA. Is there a specific format I should follow, or should I just attatch it as a Word document?
Thanks so much,
Erik Pauls
********************************************************* Cretaceous liability: Tyrannosaurus wrecks. Desmostylus: one weird mammal.
Erik Pauls P.O. Box 4960 Berkeley CA 94704 erik-at-uclink.berkeley.edu (510) 528-1945
I have been discussing an immunolabeling problem with Geoff McAuliffe. The original post was on the Microscopy Listserver a few days ago. The basic problem is one of double immunolabeling with mouse monoclonals.
The following message gives additional details on the labeling procedure. Since the original post, I have seen one response that suggested introducing a saturating step with fab fragments after the first labeling run.
If you have any suggestions or thoughts on other things to try, please correspond directly with Geoff at the address in the header.
Many thanks, and greetings from sunny (and HOT!) Arizona.
Bob Chiovetti (RCHIOVETTI-at-aol.com) --------------------- Forwarded message:
Dear Bob:
Thanks for responding to my querry. I should have been more specific re the objects of my study. Mouse CNS, light microscopy, paraformaldehyde fixation, 20 micron frozen sections, stained free-floating with mouse monoclonal anti Mac-1 (macrophages and microglia) with a Vector ABC Elite kit (biotinylated secondary, etc.). Mount sections on slides stain with mouse monoclonal anti-bromodeoxyuridine after trypsin unmasking (hence mounting on slides) also with a Vector ABC Elite kit. Done individualy the results are fine but when combined the staining with the second AB is greatly reduced and very spotty. The trypsin is essential after formalin fixation, and Mac1 does not survive any fixation that allows omission of trypsin (alcohol). I don't think I can do the Bromodeoxyridine first since the trypsin will digest the cell surface marker Mac-1 detects. Hot buffer antigen retrieval does not give good results with bromodeoxyuridine. If I use a rabbit polyclonal for GFAP at the first antibody I get beautiful double staining but of two seperate populations of cells so I am not sure if my problem is trying to use two mouse monclonals to stain two things in one cell or two things on one section. I am getting the idea that the secondaries are the problem? so perhaps one or both primaries linked to gold? I do need to see two colors, though. Thanks in advance!
Geoff -- *************************************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane Piscataway, NJ 08854 voice: (908)-235-4583; fax -4029 e-mail: mcauliff-at-umdnj.edu ***************************************************************
We are setting up a fluorescence microscopy lab. Would like info on insect tissue preps. lisa & vickie Vickie A. Kimler, Ph.D. Biology and Allied Health Department Mercyhurst College Erie, PA 16546 1-814-824-2169
} Our Safety officer is concerned about us decanting the liquid nitrogen } from our dewar into our coldstage using a styrofoam cup. Does anyone } have any suggestions on where we might purchase a cryo ladle of some } sort. The cryo gloves that we have, are too cumbersome when using } such a small styrofoam cup for the small quantity that we decant at a } time. } } Any suggestions would be greatly appreciated. } } Susan
This subject has been discussed a number of times before - it seems to be a common experience that safety officers don't have much/any experience of LN2 and tend to come up with all sorts of concerns which are generally wrong.
For example, gloves - they are cumbersome and make handling difficult, so you are more likely to spill the LN2. If it goes inside the gloves, you're in real trouble. Whereas some LN2 spilling on to bare skin will cause no damage at all.
First I would ask your safety officer to clarify precisely the concern about the way you are currently handling the LN2. As I say, a small spill on to bare skin will cause no damage - if you slowly pour several litres over your hands, then you might get a burn.
I would also bring up the question of shoes. If you are wearing shoes and spill LN2 into the shoe, you'll get a nasty burn. If you are not wearing shoes or socks, there will be no problem (unless you insist on standing in a puddle of LN2). So, logically, when handling LN2, all shoes (and socks) should be removed:) From personal experience, I would even suggest that handling LN2 is best done completely naked - if it spills on to your clothing, it'll get held against the skin and cause a burn.
In your case, if the safety officer insists, I'd get a strip of aluminium and bend it around to make a handle with a loop at the end where the styrofoam cup can sit.
I'm not against safety officers - it is an important job which needs doing. But enforcement of regulations, possibly written to cover rather different circumstances, by people who don't fully understand the real risks (for example, with LN2 large amounts of N2 gas are generated - has proper ventilation been considered) is not the right approach - it leads to people ignoring safety officers and saftey rules.
I'm not familiar with the kits you're using, however it appears that you're staining with a mouse primary followed by an anti mouse reagent followed by another mouse primary, is that right? If it is then there's a likelihood that the free arm/s of the second layer attached to the cells via the first primary are capturing the second primary when you put it in. You can prevent this cross-linking by using a FAB second layer, or more cheaply, if the second primary is directly conjugated (or biotinylated), you can block the spare arms of the antimouse by adding normal mouse serum after the first second layer and before the second primary.
Ray
At 8:33 pm -0400 3/6/97, RCHIOVETTI-at-aol.com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Ray Hicks ________________________________________________________________________ |University of Cambridge |Tel 01223 330149 | |Department of Medicine |Fax 01223 336846 | |Level 5, Addenbrookes Hospital |e-mail {rh208-at-cus.cam.ac.uk} | |Hills Road Cambridge |Web http://facsmac.med.cam.ac.uk | |CB2 |ftp server ftp://131.111.80.78 | |UK | | |_________________________________|_____________________________________|
On Wed, 4 Jun 1997 08:09:03 +0100 Larry Stoter {LPS-at-teknesis.demon.co.uk} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Our Safety officer is concerned about us decanting the liquid nitrogen } } from our dewar into our coldstage using a styrofoam cup. Does anyone } } have any suggestions on where we might purchase a cryo ladle of some } } sort. The cryo gloves that we have, are too cumbersome when using } } such a small styrofoam cup for the small quantity that we decant at a } } time.
} This subject has been discussed a number of times before - it seems to be a } common experience that safety officers don't have much/any experience of } LN2 and tend to come up with all sorts of concerns which are generally } wrong. } } For example, gloves - they are cumbersome and make handling difficult, so } you are more likely to spill the LN2. If it goes inside the gloves, you're } in real trouble. Whereas some LN2 spilling on to bare skin will cause no } damage at all. } } First I would ask your safety officer to clarify precisely the concern } about the way you are currently handling the LN2. As I say, a small spill } on to bare skin will cause no damage - if you slowly pour several litres } over your hands, then you might get a burn. } } I would also bring up the question of shoes. If you are wearing shoes and } spill LN2 into the shoe, you'll get a nasty burn. If you are not wearing } shoes or socks, there will be no problem (unless you insist on standing in } a puddle of LN2). So, logically, when handling LN2, all shoes (and socks) } should be removed:) From personal experience, I would even suggest that } handling LN2 is best done completely naked - if it spills on to your } clothing, it'll get held against the skin and cause a burn. } } In your case, if the safety officer insists, I'd get a strip of aluminium } and bend it around to make a handle with a loop at the end where the } styrofoam cup can sit. } } I'm not against safety officers - it is an important job which needs doing. } But enforcement of regulations, possibly written to cover rather different } circumstances, by people who don't fully understand the real risks (for } example, with LN2 large amounts of N2 gas are generated - has proper } ventilation been considered) is not the right approach - it leads to people } ignoring safety officers and saftey rules. } } Regards, } Larry Stoter } } I have the misfortune to be the safety officer in my deparment and I would like to endorse just about everything that Larry Stoter has to say about the handling of small volumes of liquid nitrogen. Eye protection in the form of goggles or a face shield is highly desirable, so total nudity is not a good idea.
Regards, Eric Lachowski
---------------------- Dr Eric E. Lachowski University of Aberdeen Department of Chemistry Meston Walk Old Aberdeen AB24 3UE +44 1224 272934 e.lachowski-at-abdn.ac.uk
On the subject of LN2, does anyone know the correct first aid treatment for a burn from this substance?
Normally, one would use cold water to cool a (heat) burn and prevent further tissue damage. But that seems inappropriate somehow...Hot water?
Perhaps in view of the known and imagined dangers and the lack of medical information re-treatment we should all wear a full immersion suit and breathing aparatus to enter any building where LN2 is stored. -- Anthony James Bentley Surface Data Scientific Instrumentation and Software Web site http:\\www.surface.demon.co.uk
I am pleased to see the thread on liquid nitrogen safety. Our lab does a lot of cryoTEM so we use lots of liquid nitrogen. Like Larry Stoter, we prefer no gloves and open safety glasses so any liquid nitrogen that might bounce onto the skin can escape easily. I was able to convince our safety officer that this made sense. We have one exception: we handle glass wide-mouth dewars (often with steel jackets.) We had one implode and send a shower of glass through the lab with such force that fragments were embedded in a fabric chair cushion. When we fill these wide-mouth glass Dewars, we wear a full face shield. -- Best Regards, John Minter
Eastman Kodak Company Phone: (716) 722-3407 Analytical Technology Division FAX: (716) 477-3029 Room 2112 Bldg 49 Kodak Park Site email: minter-at-kodak.com Rochester, NY 14562-3712 calendar: via PROFS
It probably is a function of the crystal orientation because 111 typically etches into tetrahedral holes while 100 etches with flat bottomed holes. I think if you consult some of th eearly literature you will find the best etch.
A colleague would appreciate input on chemical etching Ge(111). He has used HF10%+HNO3 90% for Ge(001) and obtained nice thin regions around the hole, but for Ge(111) got only pinholes with this solution even when cutting the strength several times.
Any suggestions?
W. Sinkler
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Wharton Sinkler PhD Department of Materials Science and Engineering Northwestern University 2225 North Campus Drive Evanston, IL 60208-3108 tel: (847) 491-7809 fax: (847) 491-7820 email: sinkler-at-apollo.numis.nwu.edu
Liquid nitrogen burns would be treated as frostbite. There are several degrees of frostbite depending on how deep the freezing damage is. Most exposure is acute, a splash on the hand or arm. The liquid nitrogen evaporates quickly enough that it cause at the most a slight reddening of the skin similar to a sunburn. This just requires observation of the sight to see if any further blistering occurs. A more severe case would involve submersion into the liquid nitrogen. This would cause more deeper damage. You must keep the frozen appendage still, in the case of hands and fingers (toes or feet) they can be rewarmed by placing in a pan of warm (~102 F) water, or under your arm. Wrap the damaged part in a sterile dry gauze, there will be blistering-do not break open the blisters, and seek immediate medical attention. If there is any doubt about the level of injury you should seek medical attention. This information is from the National Ski Patrol-Outdoor Emergency Care, and is the method of treatment for frostbite we teach to patrolers. Linda Barthel Research Associate II Department of Anatomy and Cell Biology University of Michigan lab (313) 764-7476 fax (313) 763-1166 barthel-at-umich.edu
and National Ski Patrol Outdoor Emergency Care Instructor, Mt. Brighton Michigan.
Susan Get a very small dewar, we use a 1 liter dewar for small transfers, keeps the safety people happy. Do not use a regular vacuum bottle (Thermos) as it can break and be more dangerous then the styrofoam cup. Also the plastic cryo dewars can break we broke two so far this year. I would recommend metal.
Ask the Safety person where to get a small dewar he should have the necessary catologs with part numbers. Then the safety people know you are interested in listening to them. Ours is made by Aladdin Ind.
our 4 liter is made by Union Carbide. Your LN2 supplier may also have these.
Keith Collins Albany Research Center Department of Energy Albany, Oregon collins-at-alrc.doe.gov
One person's two-pennyworth re. liquid nitrogen, cryogens etc:
I would personally be far more concerned about boiling water than boiling LN2, bearing in mind things already said on the list about the hazards of LN2 splashes on bare skin. Those who know me or who have attended the Seefeld-in-Tirol cryo-workshops will have seen my 'drinking' of the substance.
It is true that safety officers normally know very little about the realities of the LN2 and its handling, but they have a serious job to do (I know, I am also one!).
There is a huge difference between primary coolants i.e. those liquids which boil at very low temperatures (LN2 and LHe) and secondary coolants which have to be cooled by LN2 such as liquefied ethane and liquefied propane.
The secondary coolants will stick to the skin/eyes and boil off at c. -80 (ethane) and c. -40 (propane), in other words your body has to heat them up e.g. 150 degrees C in the case of propane to turn it into a gas. As body temperature is about 40 degrees C, this becomes a problem. With these, this boy does take the precautions (full face mask while liquefying and disposing), but with 'thinnish' leather gardening gloves so as to retain handling sensitivity (otherwise you are into a dangerous practice!). The thick cryo-gloves in this lab are retained solely for handling the delivery hose on the big LN2 storage tank.
A far greater risk from using LN2 is that of ASPHYXIATION resulting from poor ventilation, bearing in mind that each litre of LN2 forms approx 690 litres of gaseous N2 depending on ambient temperatures. This does tend to dilute the 18% (?) oxygen concentration in normal air somewhat. In this respect, I know of one very close encounter of the terminal kind.
In these situations I often cite "Safety consideration regarding the use of propane and other liquefied gases as coolants for rapid freezing purposes" by KP Ryan & MI Liddicoat (1987) J. Microsc. 147, 337-340.
Claimer - I wrote it.
With best wishes - Keith Ryan
++++++++++++++++++++++++++++++++++++++++++++++++++ Keith Ryan B.Sc., Ph.D., C.Biol., M.I.Biol. Plymouth Marine Laboratory, Citadel Hill, Plymouth, Devon PL1 2PB, England
My approach to this has been to be sure there are lots of open safety glasses around and to use one of those plastic beakers. You can get them with handles or, what we've done, is to save those handles from broken coffee carafes. They will last years or until some moron slams them down on a hard surface whilst cold. I've seen people trying to carefully decant lN2 from a wide mouth Dewar and it looks less safe. I've also seen people demonstrate the relative safety of lN2 by quickly dunking and removing their hand from a Dewar. It might convince your Safety Officer of something...
cheers, John Heckman Michigan State University Center for Electron Optics
Our Safety officer is concerned about us decanting the liquid nitrogen } from our dewar into our coldstage using a styrofoam cup. Does anyone } have any suggestions on where we might purchase a cryo ladle of some } sort. The cryo gloves that we have, are too cumbersome when using } such a small styrofoam cup for the small quantity that we decant at a } time. } } Any suggestions would be greatly appreciated. } } Susan } } } Susan Carbyn } Atlantic Food and Horticulture Research Centre } Agriculture and Agri-Food Canada } Kentville, Nova Scotia B4N 1J5 } Canada } } Phone: (902) 679-5566 } Fax: (902) 679-2311 } } E-mail: carbyns-at-em.agr.ca
Does anyone have on-line sign-up for equipment time? I remember this was discussed here ages ago, but I haven't heard about it recently. We are thinking about going to such a system, and would like to hear pros/cons/horror stories/suggestions.
........."Those who know me or who have attended the Seefeld-in-Tirol cryo-workshops will have seen my 'drinking' of the substance. "......
Yes, indeed I do recall the 'smoking ears and nose of Keith' during the workshops.
I have been using this substance since 1964 in large quantities. The best approach is common sense, that is what I have always demonstrated and taught to the many new comers in the exiting field of cryo applications.
Beware of plastic funnels. For several years I used a high density polyethelene funnel during some LN2 transfers. One day while being used, the HDPE exploded without warning. Shards of funnel flew all about the lab. Never did an examination to determine the cause, but suspect that over the years, micro-cracks had developed and at some point the thermally induced stress/strain was more than the (remaining) cold embrittled HDPE could stand.
I would suggest using funnels of teflon or stainless steel. ... Or none at all as is now my practice- not difficult to clean-up the spills {g} .
Responding to the message of {199706041753.NAA27849-at-phage.cshl.org} from howard-at-cshl.org (Tamara Howard): } } Does anyone have on-line sign-up for equipment time? I remember this was } discussed here ages ago, but I haven't heard about it recently. We are } thinking about going to such a system, and would like to hear } pros/cons/horror stories/suggestions. } yes, we have a web-based sign up based on our FileMaker Pro database. You can see what it looks like from our web site at URL http://resolution.umn.edu.
(It is linked as "Instrument sign-up") You will not be able to sign up (or even see the actual sign-up form) unless you have a username and ciode number here. This is a useful security feature to prevent millions on random surfers from booking all the time on our instruments.
I am currently trying to migrate to a less me-specific platform than the kludgy applescript that now does all the hard work.
This system has grown over the years and was not instantly developed as the giant behemoth it now is.
You will need to think about what size blocks of time you want, whether people can cancel their time and when, or whether they can delete other peoples time. I know it shouldn't be necessary, but have had it proved otherwise :(
Stuart
__________________ Stuart McKernan stuartm-at-tc.umn.edu Microscopy Specialist CIE Characterization Facility, University of Minnesota Phone: (612) 626-7594 100 Union Street S. E., Minneapolis, MN 55455 Lab: (612) 624-6590
To all list members on behalf of Richard Easingwood:
} To: richard.lander } From: Richard Easingwood {richard.easingwood-at-stonebow.otago.ac.nz} } Subject: Glutaraldehyde: safe limits
I originally posted this message to a safety listserver where there is a discussion running regarding safe limits for glutaraldehyde and glutaraldehyde monitoring (so it starts in mid 'discussion'). Somebody suggested I post this on this list as well, I think it is probably appropriate to anyone using GA, and I would certainly be interested to hear any feedback. From a personal point of view I'd be particularly interested to hear from anyone who thinks they've become sensitised to GA and what steps they take to minimise exposure.
} } I too am very skeptical about the maximum 'safe' exposure limit of 0.2ppm, } I am also doubtful of the use of the Glutaraldemeter (mentioned in one of } the previous replies in this discussion) for measuring safe limits as a } result. I should mention at this point that I have developed a sensitivity } to glutaraldehyde (GA), probably as a result of a small number of slight } exposures to this chemical over the last 6 years in my field of electron } microscopy. I know that if I exposed myself to air contaminated to 0.2ppm } (as measured by the Glutaraldemeter) it would be dramatically } debilitating for me - the 'steel band' goes around my chest, my energy } levels drop to nothing and I get depressed (common symptoms for this } ailment) The depression is so acute it is obviously something chemically } induced, as easy to pin point the cause as are jitters after too much } coffee. Takes about 12 hours before I feel normal again. } Now, I realise that most people suffer no such symptoms upon exposure but } neither did I for the first 5 years and I was very careful. Glutaraldehyde } was known to be hazardous when I first started using it in 1990 and this } was emphasized to me. In all the time I've used it I've actually only } smelt it a handful of times. Now I can only use our preparation lab when a } suitable mask, if I get a whiff of GA its too late for me. } } Yesterday we measured (using a Glutaraldemeter) the GA concentrations in } the air during various laboratory procedures (some very sloppy on purpose) } and the levels at no stage went higher than 0.15ppm. The smell of GA was } very strong during some of these as judged by two other volunteers (not } me, I wore my mask). Our OSH guidelines state that the odour threshold for } GA is about 0.04ppm which does not tally with our measurements made with } the meter, the smell was gauged as 'very strong' when the meter read below } this level. The meter was recalibrated before the tests and checked again } immediately afterwards. I can send a report to anyone interested. } I should say I have no desire to run down the Glutaraldemeter. We were } using it close to its limit of sensitivity afterall. I think badges fall } into the same category - they are probably fine if you want to ensure you } don't go over the official limit, the problem is that I think the limit is } too high. } } In conclusion I would say that the official safe maximum peak exposure } level of 0.2ppm is too high if you want to avoid the risk of becoming } sensitised to GA (and all the inconvenience that brings). I would say that } if you can smell it, the level of GA in the air is too high. I know that } this probably sounds hopelessly impractical for those working in Hospitals } where GA is in widespread use but I believe that we are only starting to } appreciate how dangerous GA is. } } Regards, } Richard } }
Richard Easingwood South Campus Electron Microscope Unit School of Medical Sciences University of Otago PO Box 913 Dunedin NEW ZEALAND
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I would also bring up the question of shoes. If you are wearing shoes and } spill LN2 into the shoe, you'll get a nasty burn. If you are not wearing } shoes or socks, there will be no problem (unless you insist on standing in } a puddle of LN2). So, logically, when handling LN2, all shoes (and socks) } should be removed:) From personal experience, I would even suggest that } handling LN2 is best done completely naked - if it spills on to your } clothing, it'll get held against the skin and cause a burn. } I agree with your comment about shoes, after cycling to work on a very wet night and immeadiately going to fetch an evening's supply of LN2, I found I had to fill the tipping 25l dewar from the pressurised 100l one first. This involves clouds of vapour pouring onto the floor for several minutes and I found my feet getting cold as my damp shoes started to freeze. Bare feet were safer, but maybe if I polished the shoes more often they wouldn't have got so saturated.
Alasdair Preston Dept of Mat Sci Met Cambridge Univ Pembroke Street Cambridge UK yx10000-at-cus.cam.ac.uk
} } Our Safety officer is concerned about us decanting the liquid nitrogen } } from our dewar into our coldstage using a styrofoam cup. Does anyone } } have any suggestions on where we might purchase a cryo ladle of some } } sort. The cryo gloves that we have, are too cumbersome when using } } such a small styrofoam cup for the small quantity that we decant at a } } time. } } } } Any suggestions would be greatly appreciated. } } } } Susan _______________________________________________________ Prabath Perera, Ph.D. SWT Department of Physics Research Associate San Marcos, TX 78666 Email: pp14-at-swt.edu Phone: 512/245-2131 http://www.swt.edu/~pp14/ Fax: 512/245-8233
PRINCIPAL DUTIES. Provide technical assistance to projects which focus on receptor mediated gene transfer into human cultured cells. Cut routinely gray serial sections from the embedded cultured cell monolayers followed by immunolabeling and in situ hybridization under the guidance of Associate Scientist. The projects are described on WWW: {smaller}
{/smaller} {/fontfamily} EXPERIENCE REQUIRED. Thorough hands-on experience with: a. Reichert, RMC, or PorterBlum microtomes, b. diamond, glass knives, platinum coated-glass knives; c. various embedments epon, araldite, lowicryl K4M and HM20, spurr required. Experience with embedding procedures including low temperature UV polymerization is welcome, but not essential. Ability to cut hydrated rapidly frozen or sucrose infused frozen samples is an advantage.
PROFESSIONAL OPPORTUNITIES. This job provides an opportunity to become involved in cell culture preparation for the cutting edge ultrastructural imaging technology program in energy filtering transmission electron microscopy, but ultramicrotomy will be the primary responsibility. This is a research job. Flexible work hours possible. Equal opportunity employer - minorities are strongly encouraged to apply.
APPLICATION. Please apply to:
Marek Malecki,M.D.,Ph.D.
P.I. {fontfamily} {param} Times {/param}
{/fontfamily} Address: Integrated Microscopy Resource and Molecular Biology Laboratory, 1675 Observatory Drive, Madison, WI53706. {fontfamily} {param} Times {/param}
{/fontfamily} Email: malecki-at-macc.wisc.edu {fontfamily} {param} Times {/param}
List members, This message on behalf of Allan Mitchell,
Sometime ago I asked several questions on the listserver about additives to use in the closed circuit cooling systems of electron microscopes. For various reasons I wanted to revisit the additive requirements of the closed circuit cooling system on our TEM's. Below is a summary of a larger report that outlines the information I was able to obtain and the conclusions I drew.
If any one would like a copy of the full document then contact me at the email address below.
Introduction For numerous reasons a closed-circuit water cooling system is the preferred option for providing cooling water to the electron microscope. Cooling water is required by the electron microscope to cool the diffusion pumps and to keep the electronic's and column temperature stable.
A closed-circuit water cooling system is essential if the local water supply has a high chloride concentration, has floating particles, is acidic, has a water temperature that fluctuates and is uncontrollable (this potentially leads to specimen drift problems in the TEM) and / or has a water temperature that is very cold (this potentially leads to condensation problems or diffusion pumps not functioning properly in the TEM).
Closed-circuit cooling systems have many advantages which includes providing significantly better control of the cooling water temperature through 'heat-producing' equipment and are less susceptible to biological fouling (from slime and algae) and to the build up of scale deposits. Because of the small water 'top-up' requirements control of potential problems is greatly simplified.
In many areas closed systems are required by local by-laws because open systems (ie, connected to town supply) are extremely wasteful of water.
For further information about closed-circuit water cooling systems on the EM can be obtained from the following two books;
Vacuum methods in electron microscopy, by Wilbur C. Bigelow (1994). Pg 214 to 217 Volume 15 in the series Practical Methods in Electron Microscopy, editor Audrey Glauert
Design of the Electron Microscope Laboratory, by R.H. Alderson (1975). Pg 46 to 49 Volume 4 in the series Practical Methods in Electron Microscopy, editor Audrey Glauert
My problem With the installation of our new TEM (Philips CM100) I decided it was time to review the additives we had been be using in our closed-circuit water cooling systems. We have two systems, one for cooling our Akashi 002A TEM and the other system for cooling a Philips 410LS TEM and a Philips CM100 TEM. Both systems ran different additives; this was a historical situation.
My first aim was to find out a more about the need for additives in the TEM cooling system and my second aim was to settle on one additive that I could use in both of our TEM cooling systems.
There are a number of factors to be considered when choosing such an additive. Firstly, the anti-corrosion function; over the years I had heard a few horror stories about the inside of TEM lens coils "rusting' away due to the fact that no precautions had been been taken against corrosion.
Secondly, the anti-microbial requirement; stopping bugs growing in the system and clogging up filters and small water lines.
Thirdly, the additive should not be depleted to quickly nor require the regular replacement of all the water in the cooling system, the additive activity must be able to be replenished (each of our two reservoir tanks hold 160 litres of water and I didn't want to be replacing these volumes very often).
To start off I decided to seek advice from the Microscopy Listserver, it is here where I found the widest range of opinions.
Next I consulted some 'water experts'. Interestingly it turned out that those who could advice me about keeping 'bugs' out of the system could not advise me much about corrosion and vica versa.
Our Solution Through BDH I had discussions with Coalite Chemicals, the supplier of a wide range of biocides.
They recommended to me a product called Phylatol. Phylatol is an aldehyde biocide, has a pH of 7.0 and contains no metal ions. It is a broad spectrum biocide with the same activity as Panacide M but with a neutral pH. They also indicated that the pH of the water containing Phylatol could be adjusted up slightly if desired. I had indicated that I would like to use a pH of 7.5 to 8 and would like to adjust the pH with Sodium Bicarbonate.
We are going to use Phylatol at a concentration of 0.2% and make it up in filtered tap water.
We will buffer the Phylatol to pH 8.00 with Sodium Bicarbonate. This requires only a very small amount of Sodium Bicarbonate, approximately 0.3 gram in 2 litres of water.
Some other interesting points. Some other interesting points that came up in the discussions follow. The dimensions of the water channels within the electron microscope are often quite small. It is very important to exclude fibrous or particulate matter form these pipes both during assembly of the water lines and during cleaning of the reservoir tanks.
We have had personal experience of the frustration of insufficient water flow only to eventually find a lump of plumbers hemp (or something similar) lodged in a lens coil cooling line restricting the flow.
Ideally the materials of construction of a closed circuit cooling system should have smooth surfaces to resist being colonised by microorganisms. Bacteria tend to grow on a solid surface but they can not lodge onto a surface while a brisk flow of water is maintained. It is advisable to avoid areas of 'stagnant' water where possible in the reservoir. Bringing the return into the tank in such a way that promotes a swirling motion is one way of doing this.
Storage tanks should be smooth walled inside with a conical or dished bottom so that water can be drained completely at the time of water replacement .
Pumps with a small number of large vanes should be avoided and instead use a pump with a large number of small vanes, an Archimedes screw type pump being the best. This is to reduce pulsations in the water line which could potentially lead to specimen movement in the TEM.
The pump should be able to deliver a greater pressure than required as pressure drops off quickly in small lines. It is a good idea to use large lines right up to the EM.
To help eliminate pulsations it is suggested that a coil of copper pipe (with several coils) be located beside the pump and use a long run of plastic tubing rather than metal pipe to the microscope. If the problem is very serious then a rubber diaphragm system with air above the diaphragm to compress will help. If using copper tubing at all it must not connect to the microscope to prevent any earth loops.
Right angle bends in the plumbing should be avoided as these can also set up turbulence in the water which can lead to specimen movement problems.
One last but very important aspect to consider is the maintaining the correct pressure to the various pathways inside the microscope. This aspect will be the 'subject' of my next investigation.
Allan Mitchell South Campus Electron Microscope Unit School of Medical Sciences Dunedin New Zealand
We are trying to scan thinsections of rock, and are interested in obtaining a suitable scanner. The resolution should be on the order of typical 35mm film scanners ... its just that the sample isn't a typical 35mm slide. We've tried a flatbed with a transparency option, but it doesn't give the detail we want. Has anyone tried this??
Please contact me directly ...
cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
I am trying to compare the distribution of particles in histological tissues and I would like to determine the distance(s) of particles relative to their nearest neighbor. Since the particles are not of uniform size I thought that if I determined the centroids it will give me a reproducible parameter. I can determine distances by triangulation but there must be a software out there that can do this faster. Does anyone know of such a software? I would appreciate comments.
Beside a full face mask when liquefying ethane, propane etc., I do insist on goggles when people are decanting from a e.g. 25 litre dewar on trunnions into the 1-litre glass dewards in steel shells (I know, we should replace these with plastic). The reason for this is a story to do with their seals: the seals at the top ens tend to be incomplete i.e. there is a gap. The anecdote, from a witness, is that in one lab some LN2 went down the gap aand blew out the glass dewar, in pieces, with some violence. Also, glass can simply fail eventually (and our glass dewars are now 16 years old and used constantly for topping-up the EDX detectors).
Another horror story: in this lab, unbeknown to me (as manager of EM plus LN2, and also Safety), a student was told bld by someone who shopuld have known better to take some LN2 in a standard picnic Thermos with a screw top. He screwed on the top and transferred to a nearby lab. It made a very good BOMB! We were picking glass out of fish tanks for days afterwards, plus all the work of transferring fish etc. DO NOT SCREW ANY LIDS ONTO LIQUID NITROGEN CONTAINERS, FOLKS! It boils back to a gas as temperature rises and if it is in defined volume then pressure rises until something gives.
Dear All I agree with what was said in this posting, except that in my case I would have to substitute formaldehyde.
The sensitisation occurred one evening in 1972. I spent several hours over a dissecting microscope working on a fish head/brain preparation after perfusion with 4% formalin + 5% glut. in a small room with no ventilation.
About 3 am I woke up with really burning eyes. I was taken to the (fortunately local) eye hospital immediatelt only to be flushed out (not nice) and given eye drops against infection. My eyes were covered for about 4 days. I couldn't go back to contact lenses for years afterwards.
I am now overpowered by low concentrations of formaldehyde. Even 0.04 ppm(measured by a meter) seems far too much too me and my eyes inflame, my throat becomes sore and sometimes my sinuses will become irritated if I stay there too long. In our lab, it is only used in a fume cabinet/cupboard or under a strong extractor fan in one place.
I would like to know why you moved to LaB6 as in my experience the SX40 will perform pretty well with W if handled correctly. What kV are you using would be of assistance in solving your problem, that is if it is no= t the LaB6 set up? For performance - 1. Working distance less than 10mm, 5 to 8 if working at {15kV 2. Spot size must be beyond half way, probably ~60% if gun is correctly set, depends on specimen emission characteristics 3. If gun is set up correctly for W emission current should be standing current plus 100uA 4. With LaB6 if filament is correctly set you should be able to obta= in an image even at the smallest spot size, if not double check your "saturation", first peak is bright but the dip following is the highest resolution position. 5. If 4 is not possible move the filament forward, current ~ standin= g current plus 20 to 50uA 6. Remember the highest resolution will only be obtained when the hi= gh voltage tank "heat gained =3D heat lost", about 1 to 11/2 hours with the= kV on! 7. Dirty column should not be a problem if you run at 30kV but lowering the kV will make the beam more susceptible to column contamination.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Haevy metals will give more backscattered electrons than light ones. A low dislocation content and a clean surface helps also.
If you intend to do just a demo of an electron backscattering pattern EBSP or a SACP pattern , I would suggest to take the filament (W) of an old incandescence bulb which died under normal aging. Just break the glass with a glass saw (or a hammer, after embedding it in a cloth to take care of your fingers), pick the filament. You will see nice faceted W crystals due to recristallisation. EBSP/SACP patterns present a very high contrast. Better use a spot/low voltage bulb, it will have a larger filament diameter.
Regards
Philippe-A. Buffat
__________________________________________________________________ Philippe Buffat Ecole Polytechnique Federale de Lausanne (EPFL) Centre Interdepartemental de Microscopie Electronique Address: EPFL-CIME, Batiment MX-C, CH-1015 Lausanne, Switzerland Phone: +41(21)693 29 83 Fax: +41(21)693 44 01 (Central European Time) E-mail: philippe.buffat-at-cime.uhd.epfl.ch, WWW URL http://cimewww.epfl.ch/ ______________________________ Eudora F2.1 ___________________________
I checked my "bible" and found about 80 pages of Ge etchants, not including Ge alloys. Etchants range from air to mercury nitrate.
CRC Handbook of Metal Etchants Walker, Perrin & Tarn, William H., Eds. 1991 CRC Press, Boca Raton, FL ISBN 0-8493-3623-6
1400 pages. Includes references.
------------------------------------------------ Opinions or statements expressed herein, rational or otherwise, do not necessarily reflect those of my employer.
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 Phone: (508) 750-1717 E-mail: crossman-at-osi.sylvania.com
Our web sites: www.sylvania.com www.siemens.com --
hi Folks, we would like to use xcosm to deconvolve some z series data sets, however we need to strip the data and present it as a raw series to the package, we are able to generate tifs, picts, or other more proprietary formats (BDS DAT files, or Imagespace series) from the micrscopes but need to prepare the data for xcosm, I would like to hear how others have gone about solving this problem Tx
Simon C. Watkins Ph.D. Associate Professor and Director CBI University of Pittsburgh Pittsburgh PA 15261 tel:412-648-3051 fax:412-648-8330 URL http://sbic6.sbic.pitt.edu
--IMA.Boundary.943125568 Content-Type: text/plain; charset=US-ASCII Content-Transfer-Encoding: 7bit Content-Description: cc:Mail note part
Michael,
How about using the Polaroid 35 mm slide scanner with the histoslide adapter? We use it quite successfully to scan large histoloty sections at high resolution. Down side is time to scan and large file size. Check with Polariod.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We are trying to scan thinsections of rock, and are interested in obtaining a suitable scanner. The resolution should be on the order of typical 35mm film scanners ... its just that the sample isn't a typical 35mm slide. We've tried a flatbed with a transparency option, but it doesn't give the detail we want. Has anyone tried this??
Please contact me directly ...
cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
(IMA Internet Exchange 2.1 Enterprise) id 0010A583; Wed, 4 Jun 97 19:13:02 -0500 Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by ns1.baxter.com (8.8.0/8.8.0) with SMTP id TAA19835; Wed, 4 Jun 1997 19:21:30 -0500 (CDT) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id QAA04873 for dist-Microscopy; Wed, 4 Jun 1997 16:57:05 -0500 Received: from aaem.amc.anl.gov (aaem.amc.anl.gov [146.139.72.3]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id QAA04870 for {Microscopy-at-sparc5.microscopy.com} ; Wed, 4 Jun 1997 16:57:04 -0500 Message-ID: {3395E48A.C31C36B2-at-darkwing.uoregon.edu}
On Wed, 4 Jun 1997, Corazon D. Bucana wrote:
} I am trying to compare the distribution of particles in histological tissues } and I would like to determine the distance(s) of particles relative to their } nearest neighbor. Since the particles are not of uniform size I thought } that if I determined the centroids it will give me a reproducible parameter.
It will but the data will be quite different from what you might get from measuring the shortest distance between the outer perimeters of the objects. That, of course, is a much more tedious process. (We have faced some of the these issues before since we are interested in the size and spatial distribution of myofibrils in striated muscle.) One way to assess the distances between object, as opposed to that between centroids, is to use stereological measurements of the spaces between the objects at varying angles of testing.
PRINCIPAL DUTIES. Provide technical assistance to projects which focus on receptor mediated gene transfer into human cultured cells. Maintain human cell culture stocks under the guidance of Associate Scientist. Record microscopic images of the cultured cells. This job shall also include preparation of the media, freezing and thawing of stocks, sterilization of glassware, keeping records, maintaining supplies, general lab clean-up. The projects are described on WWW: {smaller}
EXPERIENCE REQUIRED. Hands-on experience with sterile work with cell cultures under the laminar flow is required.
PROFESSIONAL OPPORTUNITIES. There is an opportunity to become involved in preparation of cells for modern imaging technologies e.g. two-photon excitation, confocal, low-dose fluorescence with deconvolution of images, rapid cryoimmobilization, in situ PCR, immunolabeling, eftem, but maintaining cell cultures will be the primary responsibility. This is a research job. Flexible work hours possible. Equal opportunity employer - minorities are strongly encouraged to apply. {fontfamily} {param} Times {/param}
{/fontfamily} APPLICATIONS. Job will become available on the15th of June1997. Applications will be accepted until position is filled. Candidates are encouraged to submit applications, CVs, names of referees, and/or letters of recommendations to: Dr. Marek Malecki, P.I. {fontfamily} {param} Times {/param}
{/fontfamily} Address: Integrated Microscopy Resource and Molecular Biology Laboratory, rm159, 1675 Observatory Drive, Madison, WI53706. {fontfamily} {param} Times {/param}
{/fontfamily} Email: malecki-at-macc.wisc.edu {fontfamily} {param} Times {/param}
} On Wed, 4 Jun 1997, Corazon D. Bucana wrote: } } } I am trying to compare the distribution of particles in histological tissues } } and I would like to determine the distance(s) of particles relative to their } } nearest neighbor. Since the particles are not of uniform size I thought } } that if I determined the centroids it will give me a reproducible parameter.
If the distance you really want is the separation between centroids of the sections in the plane, you can get if from their centroids. If what you really want is the 3D distance between the surfaces of the particles, you can get it by drawing random lines (which are random in 3D if your section was random) on the image and measuring the length of the intercept lines across the inter-particle background. This is a tad complicated sounding, but actually rather simple. Bear with me Each intercept length we will call L. You want to determine the average value of the reciprocal, 1/L. The average value of 1/L is two-thirds of the value of 1/T where T is the actual mean interparticle spacing (surface to surface). The proof is geometrical and somewhat arcane, but well known [;-)] in the stereological literature.
One of our customers is inquiring about the possibilities for time lapse video recording and motion analysis. If you have a system that you are happy with, I would appreciate hearing about it.
Vendors are also encouraged to respond, but *please,* no sales pitches on the list. Post to the list only if the material is of an educational nature or involves new technology. Otherwise, please respond directly to me off-list.
The questions we need answered are as follows. This would be for a system which is used on an inverted microscope for cell culture:
1. Option #1 would be to equip an existing inverted microscope for time-lapse video recording. This scope already has a CCD camera on it. What are the possibilities for time-lapse VCRs?
2. Option #2 (I'm assuming) would be to interface the camera to a computer with a framegrabber board that runs motion analysis software. What is the state of the art here? Also, what are the requirements for computing horsepower?
3. Option #3: Could we combine #1 and #2? In other words, could we acquire images with a time-lapse VCR, then later send the images to the computer for motion analysis?
Your thoughts and opinions would be greatly appreciated. Thanks!
Bob Chiovetti E. Licht Company (RCHIOVETTI-at-aol.com)
I am doing a friend a favor (no good deed ever goes unpunished). He has a backscatter detector on a Hitachi SEM but has lost most of the documentation. The unit is labeled "GW Electronics Type 113." His question is "What is the resolution of the detector (smallest change in Z detectable) and is it a function of accelerating voltage?"
I'm getting some human hepatocytes for possible DAB staining within the next couple weeks. I have no experience with human hepatocytes or DAB. Can anyone recommend an appropriate fixative and buffer? Also, how long can I store these and they'll be o.k. for DAB staining? Should I store them in buffer after a period of fixation? Any help / advice would be appreciated.
Marcia Pitzenberger Merck & Co., Inc. P.O.Box 4, WP45-251 West Point, Pa 19286-0004 (215)652-9767 marcia_pitzenberger-at-merck.com
Frank wrote, } } I am doing a friend a favor (no good deed ever goes } unpunished). He has a backscatter detector on a Hitachi } SEM but has lost most of the documentation. The unit is } labeled "GW Electronics Type 113." His question is "What } is the resolution of the detector (smallest change in Z } detectable) and is it a function of accelerating voltage?" } Dear Frank, Several conditions affect the smallest Z detectable. They are KV, atomic number, tilt of the stage, beam current and scan rate. If you would like, I can fax to you part of the manual for the type 30a/113a backscattered electron detector. It is about 18 pages long. Call or e-mail me.
Gregory Rudomen University Microscopy Imaging Center S.U.N.Y. Stony Brook Greg-at-UMIC.SUNYSB.EDU 516-444-3126 *The opinions expressed above are my own and are not necessarily shared by the Microscopy Center*
Post-Doctoral Position in Analytical Electron Microscopy
The Structural Materials Research Section at the Pacific Northwest National Laboratory (PNNL) has an opening in the area of analytical electron microscopy. This is a one-year post doctoral position with the possibility of an extension for a second year. The research will involve high resolution compositional measurements at grain boundaries and interfaces as well as some microstructural analysis. Materials are primarily Al-based alloys and the work would support programs in interfacial deformation, recrystallization and stress corrosion cracking. The candidate is expected to have a Ph.D. in materials science, physics or a related discipline and expertise in utilizing EDS and/or PEELS with a transmission electron microscope for quantitative compositional analysis. Expertise in operation of a FEG-TEM is also beneficial.
PNNL is a national laboratory located on the Columbia River in SE Washington state. We have a well-equipped microscopy lab featuring a JEOL 2010F (Oxford EDS system and Gatan PEELS) in addition to several other microscopes (JEOL 1200, Philips 400, VG HB501) and associated sample preparation facilities.
For consideration, please submit a resume (including references) and publication list by Aug. 1, 1997 to:
Dr. John Vetrano Pacific Northwest National Laboratory MSIN P8-16 P.O. Box 999 Richland, WA 99352
When an instrument is operating the transformers and rectifiers in the HT=
tank generate heat, Whilst they are warming up the high voltage drifts t= o a level that is visible as a focus change on the screen over a few minute= s at 30,000X plus, or on a photograph at 15,000X with a 30secs plus exposur= e. Give the tank time to warm up such that there is thermal eqilibrium and=
the stability will or should be very good. One to one and a half hours should be sufficient in a SEM depending on kV and model. ALL electron optical instruments suffer from this problem its physics!
This is a bigger problem with high resolution TEM where a 100kV high voltage tank may take up to two hours to stabilise. A freon, or similar media, filled HT tank warms, even with a 100kV machine to give stability = in about 45 minutes!
Most people confuse focus drift with a final lens problem in the SEM wher= e as it is more likely high voltage change resulting in focus change.
} I need a lens with a "C"-mount for a Sony Model XC-77 analog video camera. } Can anyone recommend a source? A macro lens would probably work the best } for my application (image analysis). } } Thank you.
Hi Delilah,
What I have done is fit an adaptor mount to the video camera which enables me to attach an ordinary 35mm-camera lens. The brand of adaptor I have is Rowi ('cos that's all that was available at the time) but there are probably other manufacturers. The Rowi works fine though.
In my case I have attached a Nikon 50mm macro lens to do image analysis on large histological sections which are on a light box about 2 feet away. The adaptor works fine in these circumstances, but because the lens is not mounted directly on the camera (i.e. is separated from it by the adaptor "tube") it is not possible to focus on objects far away. In other words it is optically like fitting bellows or extension tubes between a camera and its lens.
In case you don't know what an adaptor is, it is simply a black tube; one end screws into the C-mount, the other end has a bayonet fitting as found on advanced 35mm cameras to which a 35mm camera lens can be attached - in my case it is a Nikon-type bayonet mount. Any large photo or video store should be able to help you.
Regards
Stephen Edgar
Electron Microscope Unit, Pathology Department School of Medicine University of Auckland Private Bag 92019 Auckland New Zealand
I agree with the previous posts regarding the supposed "safe limits" as being too high. I too have become sensitized from years of exposure to low levels. If I catch even a whiff of glut. my throat becomes irritated and I cough for a couple of days. I insist that it always be dealt with in a fume hood, no exceptions. Just my two cents worth.
cheers ed
Edward J. Basgall, PhD The Pennsylvania State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618
Related to the cryo ladel and safety thread, does anyone know of a source for a dewar with a pouring spout?
We keep our LN2 in either a 5L or 10L dewar. Expecially for the SEM EDAX system, trying to pour from the 5L into the detector's vessel up near the ceiling is a backbreaking job. So we are left filling a very small (500 ml?) dewar about a million times going back and forth to finally fill the detector. As if this wasn't tedious enough even the small dewar doesn't pour well. (We've avoided using plastics such as polypropylene tripour beakers after having an exploding funnel episode of our own.) What would be great would be a ~1L dewar, kind of shortish, with a pouring spout. Anyone know of such a beast??
Thanks, Karen
-- Karen Zaruba kszaruba-at-mmm.com 3M Company, 3M Center Bldg. 270-1S-01 (612)737-2971 St. Paul, MN 55144 fax: 736-1519 "The opinions stated above are my own, not necessarily 3M's"
} } I need a lens with a "C"-mount for a Sony Model XC-77 analog video camera. } } What I have done is fit an adaptor mount to the video camera which } enables me to attach an ordinary 35mm-camera lens. snip } In my case I have attached a Nikon 50mm macro lens to do image } analysis on large histological sections which are on a light box } about 2 feet away.
I use the same setup on an XC-77 and it works very well. If you have no local supplier for the adapter, you can contact Edmund Scientific. They have a website and a (US) 800 number.
Let me add my voice-in-the-wilderness to this issue. I'm slightly sensitive to glut, not so bad I can't use it, but gloves and fume hoods are a *must*. Anyone not sensitized should demand them to prevent getting sensitized. Same for formaldehyde.
The problem doesn't end with these compounds, though--I've become sensitized to cacodylate and MS-222 (an anesthetic), and others have remarked on becoming sensitized to embedding resins. I would think the chronic exposure limits are too high generally, but in all cases gloves and fume hoods should be used more than they are.
The question is, what is to be done with anatomy classes? I've taken to recommending that any women who are pregant or working on it to seriously consider dropping the course.
Phil
P.S. There was a thread awhile ago on what gloves to use with what chemicals--was there a definitive list that came out of that discussion? I know glut goes through latex like the glove isn't there. P
} Sic Hoc Legere Scis Nimium Eruditionis Habes { Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217) 355-1143 oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
Most BSE detectors are said to be able to resolve about a tenth of an atomic number, particularly at the light element end of the spectrum. Ad= d a multi channel analiser and even better performance is possible.
As the accelerating voltage is decreased the efficiency of the detector will decrease due to detector conversion efficiency dropping along with specimen signal levels.
If the detector is from the 80s one would not expect to obtain results fr= om an accelerating voltage of much less than 15kV with a WD of 15 to 20 mm (detector specimen distance of 10 to 15mm).
If the detector comes from the early 90s we would expect to be able to wo= rk down to 5kV provided the electron gun was well adjusted giving us a good level of probe current.
This is for the jury-riggers out there and is definitely not UL or OSHA approved, but I think it is less hazard prone than a lot of LN2 ladling and pouring.
I have a 20 liter dewar for which I built a simple system to transfer LN2 at floor level to an EDS dewar at a height 63 inches.
Total cost is about 10 to 20 dollars plus a few hours of assembly. It requires a source of low pressure air.
The system uses a 6 foot length of 3/8" copper tubing insulated with dense foam tube, some PVC pipe, foam insulation spray, air couplings, two O-rings and a teflon seat ball valve.
My dewar plug is made from 3 inches of PVC pipe of the same diameter as the dewar throat. I machined some 'O' ring seats along its outside for a snug fit and seal. The long outlet tube and short air inlet tube are placed through the pipe and set in place with household foam insulation in a spray can. You may need to make an armature to hold the tubing in place during the cure. Try not to remember me while you are messing with this wet sub-foam slop. The inlet tube and outlet tube should not be in contact. The outlet tube should extend to near the bottom of the dewar and the inlet tube should be cut off at the bottom of the plug. The valve is on the inlet supply side.
I hold the plug while dispensing. If high pressure air is used and no restraint applied to the plug it could be blown out of the dewar neck. This and careless overfill are the only dangers unique to the apparatus I can think of.
A simpler system could probably be built with a two holed rubber stopper and a bracket to secure it in the dewar's throat. Stopper rubber, however, might not hold the copper tubing which immediately drops to LN2 temps and would develop a lubricating layer of ice and water. Keeper sleeves could be soldered onto the copper tubing on the underside of the stopper.
Transfer is by pressurizing the sealed dewar through the valved inlet tube with about 10 pounds of compressed air. I safely transfer 7.5 liters of LN2 in about 30 seconds with no back strain or sight of the liquid. If the exit tube is just the right length one can drain the dewar to the last "drop" without tipping the vessel.
A styrofoam float with a calibrated stick long enough to poke through the dewar's throat can be used to monitor fill progress when the destination dewar is located up high.
This is for the jury-riggers out there and is definitely not UL or OSHA approved, but I think it is less hazard prone than a lot of LN2 ladling and pouring.
I have a 20 liter dewar for which I built a simple system to transfer LN2 at floor level to an EDS dewar at a height 63 inches.
Total cost is about 10 to 20 dollars plus a few hours of assembly. It requires a source of low pressure air.
The system uses a 6 foot length of 3/8" copper tubing insulated with dense foam tube, some PVC pipe, foam insulation spray, air couplings, two O-rings and a teflon seat ball valve.
My dewar plug is made from 3 inches of PVC pipe of the same diameter as the dewar throat. I machined some 'O' ring seats along its outside for a snug fit and seal. The long outlet tube and short air inlet tube are placed through the pipe and set in place with household foam insulation in a spray can. You may need to make an armature to hold the tubing in place during the cure. Try not to remember me while you are messing with this wet sub-foam slop. The inlet tube and outlet tube should not be in contact. The outlet tube should extend to near the bottom of the dewar and the inlet tube should be cut off at the bottom of the plug. The valve is on the inlet supply side.
I hold the plug while dispensing. If high pressure air is used and no restraint applied to the plug it could be blown out of the dewar neck. This and careless overfill are the only dangers unique to the apparatus I can think of.
A simpler system could probably be built with a two holed rubber stopper and a bracket to secure it in the dewar's throat. Stopper rubber, however, might not hold the copper tubing which immediately drops to LN2 temps and would develop a lubricating layer of ice and water. Keeper sleeves could be soldered onto the copper tubing on the underside of the stopper.
Transfer is by pressurizing the sealed dewar through the valved inlet tube with about 10 pounds of compressed air. I safely transfer 7.5 liters of LN2 in about 30 seconds with no back strain or sight of the liquid. If the exit tube is just the right length one can drain the dewar to the last "drop" without tipping the vessel.
A styrofoam float with a calibrated stick long enough to poke through the dewar's throat can be used to monitor fill progress when the destination dewar is located up high.
I am looking for some direction on the preparation of diverse protozoans (several common ciliates, Euglena and A. proteus) for scanning electron microscopy. My major difficulties lie in the areas of keeping them "relaxed" during fixation and getting them to adhere well to some substrate such as polycarbonate filters, PLL coated coverslips, etc. Additionally, fixation of the Amoeba is a disaster for me, although the others look OK with a glut/osmium mix.
Any tips or references would be greatly appreciated; thank you in advance.
Dick Briggs Biology Department Smith College Northampton, MA 01063
I want to do some image processing and analysis of some secondary electron images of mineral grains. These are from grain mounts rather than polished mounts and are available as 8 bit grayscale images. The grains are not separated nicely against background but rather are piled up, giving substantial overlap. However, a sufficient number of grains are not overlapped that some assessment of their properties should be possible. I am able to segement the images to a binary state (all pixels 0 or 255) but attempts to find and analyse the objects are not very successful because partially hidden grains fade into the dark background and do not segment cleanly.
Does anyone have experience of doing this kind of processing? Is it possible? If anyone can give me a list of steps to follow I'd be most grateful.
NB I'm doing the image processing/analysis in Image-Tool 1.27.
If you choose to pressurize your LN2 dewar, be careful of condensing Liq Oxygen in it. The oxygen in the pressurizing air condenses at a higher temperature than the LN2 so that it is possible to wind up with a large amount of Liquid oxygen in the dewar. Liquid oxygen is hazardous to deal with. This is particularly a problem if you never completely the dewar as the LOX can build up over time. Pressurizing with N2 gas is MUCH safer.
Henk Colijn
} } This is for the jury-riggers out there and is definitely not UL or OSHA } approved, but I think it is less hazard prone than a lot of LN2 ladling } and pouring. } } I have a 20 liter dewar for which I built a simple system to transfer } LN2 at floor level to an EDS dewar at a height 63 inches. } } Total cost is about 10 to 20 dollars plus a few hours of assembly. It } requires a source of low pressure air. } {snip}
--- On Thu, 05 Jun 1997 15:48:24 -0400 "Pitzenberger, Marcia H." {marcia_pitzenberger-at-merck.com} wrote:
} I'm getting some human hepatocytes for possible DAB staining within the } next couple weeks. I have no experience with human hepatocytes or DAB. } Can anyone recommend an appropriate fixative and buffer? Also, how long } can I store these and they'll be o.k. for DAB staining? Should I store } them in buffer after a period of fixation? Any help / advice would be } appreciated. } } Marcia Pitzenberger } Merck & Co., Inc. } P.O.Box 4, WP45-251 } West Point, Pa 19286-0004 } (215)652-9767 } marcia_pitzenberger-at-merck.com } -----------------End of Original Message-----------------
Dear Marcia,
In our lab we have routine protocols for DAB staining of human neutrophils, so you may have to adapt a little.
Fixation: 1 hour at 4-6°C (on ice) with 3% glutaraldehyde in 0.1M cacodylate buffer, pH 7.35.
They are washed 3x and stored overnight in 0.1M cacodylate 7% sucrose buffer, pH 7.35.
The sooner DAB staining occurs the better. Activity falls off such we try to have the samples stained within 2 weeks (3 at the outside). The hepatocytes may differ from that however.
The DAB staining that we do is for myeloperoxidase (0.01% H2O2 for 30 min at room temp in the dark) but it also works for the various catalase procedures. If you require any references, I may be able to provide some. Let me know.
Hope this helps,
Chuck ------------------------------------- Name: Charles Gilbert VOC:(704)355-5261 Carolinas Medical Center FAX:(704)355-8424 Dept of Pediatric Research PO Box 32861 Charlotte, NC 28232-2861
Thanks for your suggestion. Although your method may be well known in the stereological literature, it probably isn't to microscopists. It would be nice for you to furnish an appropriate citation or two.
On Thu, 5 Jun 1997 DrJohnRuss-at-aol.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } In a message dated 6/5/97 2:02:29 PM, you wrote: } } } On Wed, 4 Jun 1997, Corazon D. Bucana wrote: } } } } } I am trying to compare the distribution of particles in histological } tissues } } } and I would like to determine the distance(s) of particles relative to } their } } } nearest neighbor. Since the particles are not of uniform size I thought } } } that if I determined the centroids it will give me a reproducible } parameter. } } If the distance you really want is the separation between centroids of the } sections in the plane, you can get if from their centroids. If what you } really want is the 3D distance between the surfaces of the particles, you can } get it by drawing random lines (which are random in 3D if your section was } random) on the image and measuring the length of the intercept lines across } the inter-particle background. This is a tad complicated sounding, but } actually rather simple. Bear with me } Each intercept length we will call L. You want to determine the average value } of the reciprocal, 1/L. } The average value of 1/L is two-thirds of the value of 1/T where T is the } actual mean interparticle spacing (surface to surface). } The proof is geometrical and somewhat arcane, but well known [;-)] in the } stereological literature. } } John Russ } }
Edmund Glaser, D. Eng. Dept. Physiol. Univ. Md. School. Med. Baltimore, MD 21201 USA Ph: (410) 706-5041 Fax: (410) 706-8341
I have a customer half way across the country who would like to examine the possibly corroded surface of a large mold in a plastics-forming plant. The mold is too big to be put in the SEM and it cannot be sectioned (big $). I suggested making a plastic film replica of the surface and sending it to me.
I can't seem to find my references/instructions for making acetate (?) replicas with acetone and I want the customer to be able to do it right (we have only one chance).
Any guidance would be appreciated, even from vendors. :-)
------------------------------------------------ Opinions or statements expressed herein, rational or otherwise, do not necessarily reflect those of my employer.
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 Phone: (508) 750-1717 E-mail: crossman-at-osi.sylvania.com
Our web sites: www.sylvania.com www.siemens.com --
Advice needed, We are about to purchase a high vac metal evaporator for shadowing and carbon deposition, and would like some recommendations. We need a dependable workhorse, as this equipment will be destined for many core users. High vac (10 -7 mbar) within a reasonable time, durability and ease of use is what we want. We are currently looking at the Edwards Auto 306 (possibly with a cryo-pump, more likely with a diff pump) and Denton's DV-502A (we have between 15-20K to spend). We have been using an old Polaron which is on its last legs. All thoughts and suggesstions are greatly appreciated.
Mike Delannoy JHMI Microscopy Facility (410) 955-1365
In a message dated 6/6/97 10:04:09 AM, eglaser-at-umabnet.ab.umd.edu wrote:
} Thanks for your suggestion. Although your method may be well known in } the stereological literature, it probably isn't to microscopists. It } would be nice for you to furnish an appropriate citation or two.
H. J. Gundersen et al, 1978, J. Microscopy v.113, p.27 J. C. Russ, 1986, Practical Stereology, Plenum, New York, p. 62
I can't resist the opportunity to point out that microscopists SHOULD study the stereological literature - they are examining 2D images of sections cut through 3D structures, and if they ignore the stereological meaning of their 2D measurements (no matter how carefully done) the results will not be meaningful. End of soapbox.
Dear Roger, here at Noesis Vision Inc we have lots of experience with particle separation. We have powerful separation algorithms in Visilog, send us your images and we can do a feasibility study for you at no charge.
Regards,
At 08:41 AM 6/6/97 -0230, Roger Mason wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
---------------------------------------------------------------------------- ------------- Luc Nocente Tel: 514 345 1400 Noesis Vision Inc. Fax: 514 345 1575 e-mail: ln-at-noesisvision.com http://www.noesisvision.com 6800 Cote de Liesse, Suite 200 St-Laurent, PQ H4T 2A7,Canada ---------------------------------------------------------------------------- -------------
We use old metal coffee pots for LN2. They have straight sides, have a pour spout, and plastic insulated handles. Well-designed to keep hot coffee from your hands, they do good service with liquid nitrogen. Water will condense on the outside and freeze, but this doesn't get into your dewars if transit time is reasonable. They are aluminum and available in most hardware stores in various sizes from 0.5 to 2 or more liters for around $10. They hang up well and don't break on falling or cooling.
I also like metal soup ladles, also from the hardware store, with about half liter capacity. Cost about $4. These are spot welded and come apart after a few years of jolting about. We keep half dozen hanging over our work area so that they warm up and dry out between uses.
Jacob
Jacob Bastacky, M.D. Room 116 Donner Laboratory Lawrence Berkeley Laboratory EMail: sjbastacky-at-lbl.gov University of California Phone: (510) 486-4606 Berkeley, California 94720 Fax: (510) 486-4750
Does anyone have suggestions of software for analysing negatives taken on a TEM with negatively stained material? (I'm currently using NIH-Image, but would like to render, if possible, 3D volumes of my material)
Also, I wonder if there's any known way to estimate/calculate the height of structures from negatively stained material scans...
In message {3396DE9A.5AF-at-accessone.com} , Bart Cannon {cannonmp-at-accessone.com} writes } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
What about a source of low pressure nitrogen - like for instance a dewar of LN2? (g)
Drop a small heating coil in the dewar. -- Anthony James Bentley Surface Data Scientific Instrumentation and Software Web site http:\\www.surface.demon.co.uk
I must confess I hadn't thought about oxygen condensation.
Luckily my design uses no sparkable materials.
While on the subject I wonder how my system would produce more gas stratification that any sitting, capped dewar since the leak tight cap is only used for 30 seconds.
This inspires the question: are all capped dewars oxygen bombs???
Some LN2 can be transferred from the introduction of the transfer tube into the LN2, but not enough for a fill. The compressed air is so quick and easy I hadn't bothered to look into N2 as a pressurizing gas.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi All-
I'm looking to purchase a sample holder to use with our stereo microscope. I need it to hold samples from one eighth inch to several inches and to be adjustable (turn the sample). I've looked through all of our catalogues with no success. Any ideas?
All over our building we have an argument raging as to the correct formulation for PBS. Mainly the argument consists of opinions that the phosphate concentration should be 0.1M, and other opinions that the concentration ought to be O.OlM. (We all agree that NaCl concentration is 0.15M). There are also conflicting statements in the literature as to the composition of PBS. What we need is a thorough explanation of why one or the other concentration of phosphate salts is correct for mammalian tissue for TEM in procedures such as the DAB process. I would so much appreciate help in logically settling this argument around here. So long, Hildy
Hildy Crowley University of Denver 2101 E. Wesley Ave Denver, CO 80208
Replicas are easy to make. Cut the Acetate to size then place a drop or two of acetone on surface to be replicated. Place the acetate on surface. The acetone will soften the acetate so you will want to apply slight pressure to ensure acetate molds to surface. I use a cotton swap, but have also used my finger.
If you need to do the top surface put the acetone on the acetate.
You can by a kit from Bueller (most likely spelled wrong) that has foil on the back which allow viewing with an optical microscope.
The first replica will clean the surface, save it for analysis as in this case it will contain corrosion products.
Do more than one just incase something goes wrong. (it does for me.)
For those of you who were looking for the Leaf 45, I think that I found the site for the new improved version of it. Check this site out http://www.hyperzine.com/photokina/brem.html
Here is a short paragraph from the page.
Bremson Negative Scanner
photokinaBremson Inc. announces the debut of the versatile new Bremson 45HS Scanner, a high resolution, multi-format negative scanner designed by Leaf Inc., for exclusive manufacture and distribution by Bremson Inc. Well below the price of comparable high-end, high cost drum scanner technology, the affordability of the new 45HS Scanner is a welcome addition to commercial and other labs that specialize in digitizing negatives.
I have no interest in this company and was surfing the net looking for negative scanners. No one seemed to know precisely what happened to the Leaf 45.
If you are looking into scanners, here's a place to start.
This is a site by the Scientific Photography Lab at the University of Basel that I found that has a long list of scanners and information on them separated by formats:
http://www.foto.unibas.ch/scanners.html
Very informative if you are looking for a scanner.
Richard, I was reading my wife's e-mail and noticed your question. I am an occupational hygiene chemist and have been measuring glutaraldehyde levels in hospitals and clinics throughout Queensland for about six years, and I agree that 0.2ppm is far too high. Following representations from a number of people, myself included, Worksafe Australia have reduced the limit to 0.1ppm . I dont consider myself to be particularly sensitive to glutaraldehyde, but I know that 30mins exposure to 0.09ppm will guarantee a headache. In my experience, to avoid complaints from staff, the level should be kept below 0.03ppm if possible, although I did come across someone who became sensitised at 0.02ppm while working in an X-ray dark room. X-ray film fixer contains a small amount of glutaraldehyde some of which gets blown around the room when the film is dried. I usually use a Glutaraldemeter for the measurements, and it is quite good unless there is alcohol in the room. However, it is so obvious when the meter starts to wind off scale, that it is not a problem.
The Glutaraldemeter is good in that it gives spot measurements, other methods I have used such as 2,4DNPH tubes give an average reading over time, but are not so good where glutaraldehyde is being used sporadically.
Imust say that I havent come across an EM lab without a fume hood, and so they dont seem to have your problem.
Hope this helps.
George Lee Chemist- Occupational Hygiene Queensland Health Scientific Services PO Box 594 Archerfield 4108 Brisbane Queensland
Edmund Glaser wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Thanks for your suggestion. Although your method may be well known in } the stereological literature, it probably isn't to microscopists. It } would be nice for you to furnish an appropriate citation or two. } } On Thu, 5 } Jun 1997 DrJohnRuss-at-aol.com wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } -----------------------------------------------------------------------. } } } } } } In a message dated 6/5/97 2:02:29 PM, you wrote: } } } } } On Wed, 4 Jun 1997, Corazon D. Bucana wrote: } } } } } } } I am trying to compare the distribution of particles in histological } } tissues } } } } and I would like to determine the distance(s) of particles relative to } } their } } } } nearest neighbor. Since the particles are not of uniform size I thought } } } } that if I determined the centroids it will give me a reproducible } } parameter. } } } } If the distance you really want is the separation between centroids of the } } sections in the plane, you can get if from their centroids. If what you } } really want is the 3D distance between the surfaces of the particles, you can } } get it by drawing random lines (which are random in 3D if your section was } } random) on the image and measuring the length of the intercept lines across } } the inter-particle background. This is a tad complicated sounding, but } } actually rather simple. Bear with me } } Each intercept length we will call L. You want to determine the average value } } of the reciprocal, 1/L. } } The average value of 1/L is two-thirds of the value of 1/T where T is the } } actual mean interparticle spacing (surface to surface). } } The proof is geometrical and somewhat arcane, but well known [;-)] in the } } stereological literature. } } } } John Russ } } } } } } Edmund Glaser, D. Eng. } Dept. Physiol. } Univ. Md. School. Med. } Baltimore, MD 21201 USA } Ph: (410) 706-5041 } Fax: (410) 706-8341
I never subscribed to this bloody list so please stop all this friggen mail from comming here...
Harold, plastic replicas for SEM are generally easy to prepare. Preparation of a second stage, perhaps in Pt/C is much more tricky, but not needed for SEM. I used Bioden acetate film for many years but any thin plastic replicating material should do well. Bioden is used with acetone or methyl acetate.
Cut a piece of film material to size. Place on top- of the region for replication. Drop solvent onto plastic until its just wet all over. Apply gentle pressure to the film just after solvent has just evaporated to ensure good contact. Wait for at least 10 minutes before pulling replica off. Pull replica off and throw this into the bin. Repeat and "bin" second replica - to clean surface. Retain third and subsequent replica for use. Metallise (sputter) specimen surface before mounting and viewing.
Notes: Handle edge of replica and only with tweezers. Mark replica's specimen site like sheet film notching; when facing (emulsion or replica) cut the top right corner. When viewing in SEM reverse the polarity to gain a realistic "hills and valleys" impression. Single stage replicas are reversed. You could look at a "bin" replica, after carbon coating in backscattered mode to check any of the extracted material and also to do EDS on that. Angle shadow casting with Pt (from wire wrapped around a tungsten 'V' filament) and subsequent carbon coating sometimes enhances features. 10 to 30 degree angles of evaporation are used. Low angles are best for replicas with very little topography. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 350+ Links, MSDS ************************ http://www.proscitech.com.au } } I have a customer half way across the country who would like to examine } the possibly corroded surface of a large mold in a plastics-forming } plant. The mold is too big to be put in the SEM and it cannot be } sectioned (big $). I suggested making a plastic film replica of the } surface and sending it to me. } } I can't seem to find my references/instructions for making acetate (?) } replicas with acetone and I want the customer to be able to do it right } (we have only one chance). } } Any guidance would be appreciated, even from vendors. :-) } } ------------------------------------------------ } Opinions or statements expressed herein, rational or otherwise, do not } necessarily reflect those of my employer. } } Harold J. Crossman } OSRAM SYLVANIA INC. } Lighting Research Center } 71 Cherry Hill Dr. } Beverly, MA 01915 } Phone: (508) 750-1717 } E-mail: crossman-at-osi.sylvania.com } } Our web sites: www.sylvania.com } www.siemens.com } -- } } "Crossman, Harold" {crossman-at-osi.SYLVANIA.com} }
Hildy: What is the DAB process? The primary function of a buffer is to set and hold the pH at the desired level. 0.1M of the phosphates is normally used with mammalian tissues and the addition of salts to a fixative is usually not required. I remember that NaCl is 8.5g/litre in tissue fluids - and that translates almost exactly to your 0.15M figure. 0.1M buffer with the addition of that salt is certainly hypertonic. Soerenson developed the phosphate based buffer recipe. The addition of 8.5g NaCl (or a more complex balanced salt solution) turns that into PBS. No doubt, people have different fancies (experiences, data?) as to what strength the buffer should be. I suggest that 0.01 is low and 0.1M is on the high side for PBS. 0.01M is about the minimum effective buffer concentration for fixatives. That level without salt is commonly used for plants since they have much lower osmolarity than mammals do. Now, had I known what the DAB process is, I might have kept quiet! Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 350+ Links, MSDS ************************ http://www.proscitech.com.au } Dear Folks, } } All over our building we have an argument raging as to the correct } formulation for PBS. Mainly the argument consists of opinions that the } phosphate concentration should be 0.1M, and other opinions that the } concentration ought to be O.OlM. (We all agree that NaCl concentration } is 0.15M). There are also conflicting statements in the literature as to
} the composition of PBS. } What we need is a thorough explanation of why one or the other } concentration of phosphate salts is correct for mammalian tissue for TEM } in procedures such as the DAB process. } I would so much appreciate help in logically settling this argument around } here. } So long, } Hildy } } Hildy Crowley } University of Denver } 2101 E. Wesley Ave } Denver, CO 80208 }
Harold Crossman has raised some questions about the use of "acetate" replicas for SEM. The material is actually cellulose acetate, and it is available from several sources, including my employer, Structure Probe/SPI Supplies :-)
The use this material goes back to the days when fractography was done by TEM because SEM was not commercially available. There is, therefore, some very old literature on how to use the material, but since most of it is art, anyway, most current authors assume that the reader is generally familiar with the technique.
There are actually two purposes in using this material. Even for relatively small samples, it may be necessary to remove corrosion products in a nondestructive manner, and in my opinion the best way to do this is to use acetone-softened cellulose acetate, which will remove the corrosion product but leave what is left of the metal. I know about various "nondestructive" chemical removal agents, and I have done enough testing of these agents that I question whether they are truly nondestructive. As Harold points out, we have one chance at a "real" failure.
The second purpose is to make a replica of the cleaned fracture surface for examination in the microscope. Whichever the purpose, the technique is basically the same.
For TEM replication, historic practice has generally been to dissolve the replicating material in a suitable solvent at a concentration around 2%, put a drop on the area of interest, place a grid on the drop and go away for a while. Replicating materials with which I have some familiarity include collodion, parlodion, formvar, butvar, pioloform, poly(acrylic acid) and good old cellulose acetate; some of these material names are trademarks and should be capitalized; there is an appropriate solvent for each, and part of the "art" is selecting the proper combination. For example, water, used to dissolve poly(acrylic acid) is very suitable for some polymer surfaces but not suitable for most metals. When the solvent is completely evaporated, the grid can be picked up using tweezers, shadowed (or coated for SEM) and examined. This can work very well for TEM on the large, flat surfaces of those rare fatigue failures which wind up in textbooks, and it gives an unequivocal indication which part of the fracture surface generated which replica, but it has problems for "real" fractures in the SEM.
The basic problem is that the procedure is too good. The solution gets into every crack and crevice of the fracture surface, and the replica is "locked" to the surface so that it is impossible to remove without tearing. For practical SEM use, then, normal practice has been to use "tape".
Cellulose acetate is available from various suppliers (including SPI Supplies) as tape, a strip of material about 2.5 cm wide, from which the piece to be used is cut. Also available from some suppliers are sheet product. There tend to be different thickness preferences; all thicknesses work, but some folks like it thicker, and others like it thinner. In use, the area to be replicated is moistened with the solvent (acetone for cellulose acetate), the tape is also moistened on the side that will form the replica, and the solvent is allowed to soften the tape enough, but not too much (yes, there is some "art" here). I find that forming a shallow "cup" in the tape helps me to hold the solvent in the area of interest without making a mess. The obvious idea of dipping the tape into the solvent is a recipe for a real mess, because the next step will fail.
When the tape has softened enough, but not too much, it is carefully pressed into the drop of solvent on the surface of interest. Thorough contact of wetted surfaces is essential. Our "official" tool for pressing is the eraser on a pencil, but when the situation is serious, I use my right thumb; I suppose that my left thumb would work, but I've never tried :-) If the tape is too soft, the pressing tool will go through the tape, ruining the replica; if the tape is not soft enough, it will sit above the surface of interest and not capture its fine features. This is an art, not an "exact" science.
The next step is the most critical--go away for a while. Most "bad" replicas were destroyed by being "pulled" too quickly; it is essential that the solvent be completely evaporated, or the fine features will be left on the fracture surface. When completely dry, the replica is removed by pulling the tape upward. With the obvious exception that the tape may be attacked by the solvents in various "paint" products, the replica can be prepared like any other nonconductive SEM sample and examined at a remote location.
In actual practice, I do ten times as much replication to clean fracture surfaces as I do to examine them. The "first pull" replica from cleaning the fracture surface provides a "clean" sample of the corrosion product without any interference from the substrate, and in some cases, this is actually a better procedure than attempting to analyze a corrosion product still on the substrate.
Two disclaimers, first, SPI Supplies has an obvious interest in promoting the use of replicating products, including cellulose acetate tape, and second, replication involves the use of volatile solvents, some of which are somewhat "nasty". My first experience with TEM was making replicas using collodion dissolved in amyl acetate; not knowing anything about amyl acetate at that time, I can only observe that I developed the habit of driving 90 miles an hour on the way home after breating the stuff all day. Please be careful.
I would be happy to try to answer specific questions from my experience. The problem Harold presents is a very real one--he has only one chance to get the replicas, using untrained staff, and good replication is an art which requires experience.
Andy
Andrew W. Blackwood, Ph.D. Structure Probe, Inc. P.O. Box 656 West Chester, PA 19381-0656 Ph: 1 610 436 5400 FAX: 1 610 436 5755 e-mail: ablackwood-at-2spi.com WWW: http://www.2spi.com
Has anyone heard about a reference called "Ranking of the cathodes of Round-Robin tests 1 and 2 according to the structure data". Jernkontoret 1988.? I don't know if it is a book or a paper, neither where to find it.
In the "old" pre OSHA days we used a 10 watt resistor taped to the bottom of the transfer tube with a resistance that allowed it to be wired directly to 115 VAC. This could be plugged in for long enough to build up the pressure needed for transfer. Today the 115 VAC would be considered too hazardous.
However there is a way to make everyone happy. Get one of the many 1 amp, 12 volt in-line or wall mount power modules that are available (115 VAC in and 12 VDC 1 A out). You can use the same principal as above but now with an intrinsically safe voltage. No need for any pressurizing gas at all.
I have been offered a donation of a working Zeiss EM-6 SEM by someone who doesn't know anything more about it than its name. Can anyone tell me anything about this scope?(e.g., age, size, reliability, parts availability, resolution, ease of operation)
Is there anyone else out there who could use this donation?
thanks
steve
----------------------------------------------------------------------------- Dr. Steven Barlow Electron Microscope Facility Dept. of Biology San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619) 594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu http://www.sci.sdsu.edu/EM_Facility
One solution that we use is a set of steps. Our carpenter shop built (over built!) a set of wooden stairs, three steps high, that sits behind our SEM. We just climb the stairs with a 4 liter dewar when we need to fill the EDS dewar. That way we are about chest high with the opening of the EDS dewar and pouring is easy. It also prevents any LN2 from splashing down on us while filling. You can buy portable metal stairs in catalogs such as Grainger.
By the way we have 12 foot high ceilings in the SEM room!
Louie Kerr
At 3:46 PM -0500 6/5/97, kszaruba-at-MMM.COM wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Louie Kerr Research and Education Support Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543 508-289-7273 508-540-6902 (FAX)
Apologies to the list, but I've lost their email address, and can't find their posts in the archives.
Mizuho Shimizu from this company recently posted about a Nipkow disc confocal microscope.
Would someone who kept their address please send me their email address? Or if this person, or someone from their company is reading this, please contact me.
Thank you.
Phil
} Sic Hoc Legere Scis Nimium Eruditionis Habes { Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217) 355-1143 oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
How much would you think, might be the value of a light microscope with no own light source, several objectives (max. 60 x) and brass exterieur ? My great-grandfather as a passionate precision mechanic built it himself around 1910, and now somebody would like to buy it, but we have no idea how much is a sensible price.
} Date: Mon, 9 Jun 1997 09:54:07 -0500 } To: HILDEGARD CROWLEY {hcrowley-at-du.edu} } From: Tom Phillips {tphillips-at-biosci.mbp.missouri.edu} } Subject: Re: TEM:Help!Correct PBS formula???? } Cc: } Bcc: } X-Attachments: } "Saline" is a physiological term for 0.9% NaCl solution which is 0.154 M. This is osmotically correct for human serum. If you add a large amount of phosphate buffer (e.g., 0.1M), one would have to lower the NaCl concentration in an equivalent manner if you want to maintain osmolarity. The Gibco catalog gives two formulations for PBS on page 2.9 of their 1997 catalog. one lists 9.0 g NaCl, 0.795 g NaH2POH-7H2O and 0.144 g KH2PO4 (all per liter). this is about 3 mM Na2PO4 and 1 mM KH2PO4. the other formulation is 0.21 g KH2PO4 and 0.726 g NaH2PO4-7H20. either one causes a slight hyperosmolarity in respect to straight "saline". In osmicated tissue, osmolarity might not be important but I suspect in aldehyde fixed tissue it can be. In TEM, one has to worry about PO4 based buffers causing precipitation of Ca salts. In procedures like DAB, one has to ensure the amount of buffer capacity is not exceeded. } } } All over our building we have an argument raging as to the correct } } formulation for PBS. Mainly the argument consists of opinions that the } } phosphate concentration should be 0.1M, and other opinions that the } } concentration ought to be O.OlM. (We all agree that NaCl concentration } } is 0.15M). There are also conflicting statements in the literature as to } } the composition of PBS. } } What we need is a thorough explanation of why one or the other } } concentration of phosphate salts is correct for mammalian tissue for TEM } } in procedures such as the DAB process. } } I would so much appreciate help in logically settling this argument around } } here. } } So long, } } Hildy } } } } Hildy Crowley } } University of Denver } } 2101 E. Wesley Ave } } Denver, CO 80208 }
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
Biological Physics. Postdoctoral position at the University of Virginia. A postdoctoral position is available in the Department of Molecular Physiology and Biological Physics of the School of Medicine. The research projects in which the applicant is expected to play a major role involves electron energy loss spectroscopy and energy filtered scanning transmission electron microscopy. The position will include both development of software and instrumentation for achieving 2 to 3 nm spatial resolution compositional imaging and hands-on application of the method to significant biological problems. The laboratory has been engaged in NIH- supported research developing and applying analytical electron microscopy for 25 years through an interdisciplinary program based on the collaboration between physicists and biologists. Equipment available includes a 200kV electron microscope equipped with field emission gun (Philips CM200-FEG), a GATAN electron spectrometer adapted to our own CCD camera, a CM12 electron microscope and two energy dispersive X-ray detectors. Investigators in the program are also members of the Center for Structural Biology of the University of Virginia and have programs involving collaborations with the X-ray crystallography and atomic force microscopy groups and with molecular biologists and investigators engaged in other biological disciplines.
Candidates should have a Ph.D. in Physics, material science or engineering and be familiar with computer programming, instrumentation and, preferably, experience with electron microscopy and electron energy loss spectroscopy. Applications with biographical sketch, bibliography and the names of three references should be sent to: Dr. Andrew P. Somlyo, Department of Molecular Physiology and Biological Physics, University of Virginia, P.O. Box 10011, Charlottesville, VA 22906-0011, USA. The University of Virginia is an Equal Opportunity/Affirmative Action Employer.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear specialists,
How much would you think, might be the value of a light microscope with no own light source, several objectives (max. 60 x) and brass exterieur ? My great-grandfather as a passionate precision mechanic built it himself around 1910, and now somebody would like to buy it, but we have no idea how much is a sensible price.
I've never subscribed to this list so please stop all these mailings....
Imager wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Edmund Glaser wrote: } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } -----------------------------------------------------------------------. } } } } Thanks for your suggestion. Although your method may be well known in } } the stereological literature, it probably isn't to microscopists. It } } would be nice for you to furnish an appropriate citation or two. } } } } On Thu, 5 } } Jun 1997 DrJohnRuss-at-aol.com wrote: } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } -----------------------------------------------------------------------. } } } } } } } } } In a message dated 6/5/97 2:02:29 PM, you wrote: } } } } } } } On Wed, 4 Jun 1997, Corazon D. Bucana wrote: } } } } } } } } } I am trying to compare the distribution of particles in histological } } } tissues } } } } } and I would like to determine the distance(s) of particles relative to } } } their } } } } } nearest neighbor. Since the particles are not of uniform size I thought } } } } } that if I determined the centroids it will give me a reproducible } } } parameter. } } } } } } If the distance you really want is the separation between centroids of the } } } sections in the plane, you can get if from their centroids. If what you } } } really want is the 3D distance between the surfaces of the particles, you can } } } get it by drawing random lines (which are random in 3D if your section was } } } random) on the image and measuring the length of the intercept lines across } } } the inter-particle background. This is a tad complicated sounding, but } } } actually rather simple. Bear with me } } } Each intercept length we will call L. You want to determine the average value } } } of the reciprocal, 1/L. } } } The average value of 1/L is two-thirds of the value of 1/T where T is the } } } actual mean interparticle spacing (surface to surface). } } } The proof is geometrical and somewhat arcane, but well known [;-)] in the } } } stereological literature. } } } } } } John Russ } } } } } } } } } } Edmund Glaser, D. Eng. } } Dept. Physiol. } } Univ. Md. School. Med. } } Baltimore, MD 21201 USA } } Ph: (410) 706-5041 } } Fax: (410) 706-8341 } } I never subscribed to this bloody list so please stop all this friggen } mail from comming here...
As "News and Commentary" Editor for the journal "Microscopy and Microanalysis," I put a "Future Meetings" section in vol 3.2, p. 167, and "Short Courses" on p. 170. We allowed the input of extended announcements for events that were scheduled to occur close to the date of vol 3.2 (March into Fall) and brief notices for events occurring later (1998 onwards).
I plan to put in an updated Future Meetings and Short Courses section in vol 3.5, which has an input closing date to me of July 14 and will be mailed on September 24th.
I invite organizers of meetings and courses to send input for events occurring from October onwards. Extended listings will be considered for events in the October to February 1998 time period and shorter listings for later events. We will repeat short listings up to the date of the event but will allow only one extended listing. Submitters should plan accordingly.
Please send input via e-mail, offline. Please follow the format in vol 3.2, p. 167. Text submissions mailed to me will be typed by me on a time available basis. (Scary thought! :-) ) Send e-mail as plain ascii text. No attached word processor formatted documents please.
The journal goes to about 5,000 microscopists world-wide. International listings are welcomed.
Ron Anderson ron-anderson-at-vnet.ibm.com or, -at- IBM, Mail Stop E-40 Hopewell Jct., NY 12533 USA (see "scary thought," above)
This query was forwarded to me by a friend at Los Alamos who asked if I would post it to the Listserver. We would be very grateful if anyone can advise Joanna McKittrick at UCSD on this matter. Her e-mail address is jmckittrick-at-ucsd.edu
Many thanks in advance,
Gill Bond Dept Materials & Met. Eng. New Mexico Tech
---------- Forwarded message ----------
} I have a technical question. Do you know of any computational } way to generate microstructures? For example, if I have a 2 } phase material, know the average and SD of the grain size for } each phase and the volume %, is there a way to generate } an image???? Do you know anyone who does this? } } Also, do you know anyone doing combustion research here? } I am interested in thermodynamic programs, too. It seems } with the tremendous computational facilities at LANL there } should be a lot of work done in these areas. } } Last--what is your favorite drawing program? I have been } using McDraw (yuk) and am looking for something a little } more powerful. } } Thanks! } } Joanna } } ************************************** } Joanna McKittrick } Department of Applied Mechanics and } Engineering Sciences and } Materials Science Program } University of California at San Diego } 9500 Gilman Drive } La Jolla, CA 92093-0411 } } 619-534-5425 (phone) } 619-534-5698 (fax) } jmckittrick-at-ucsd.edu } ************************************* }
VOTING IN PROGRESS FOR IMMUNOCYTOCHEMISTRY NEWSGROUP sci.bio.immunocytochem
If you want to see a NEWSGROUP dedicated to immunohistochemistry, immunocytochemistry and other affinity labelling methods, then please do try to vote if you havn't already done so. The votetaker must collect 100 more YES than NO votes for the group to be made official. The closing date for ballots to reach the votetaker is 16th June. All you need to vote is a valid e-mail address.
The 2nd (and final) CFV, or CALL FOR VOTES was posted to "news.announce.newgroups" and "news.groups" on the 6th June. Voting instructions and the ballot can be found at the end of the CFV.
Some people are having trouble accessing the CFV or "Call For Votes" via the Usenet Newsgroups. The VOTETAKER, David Bostwick, will SEND YOU the CFV (with voting instructions and the ballot) if you e-mail him bostwick-at-cas.chemistry.gatech.edu
Alternatively, you can go the the FTP site: ftp://ftp.isc.org/pub/usenet/news.announce.newgroups/sci/sci.bio.im munocytochem where all the old "Request For Discussions" are stored, together with the CFV (right at the end!)
Remember, the closing date for votes to reach the votetaker is 16th June, so hurry! Please feel free to e-mail me with any questions you may have about the proposed group or voting.
Many thanks for your very practical and timely tips on making replicas. Andy Blackwood's and Jim Darley's responses were so detailed that they may have saved me a trip to Indiana! I forwarded all the messages to my customer who will be able to handle the project locally.
------------------------------------------------ Opinions or statements expressed herein, rational or otherwise, do not necessarily reflect those of my employer.
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 Phone: (508) 750-1717 E-mail: crossman-at-osi.sylvania.com
Our web sites: www.sylvania.com www.siemens.com --
I am having problems with wrinkles in Epon thin sections (gold interference colors)and would like some advice or information about getting rid of them. The specimen is ameba and has been grown on the surface of an Epon bullet coated with a collagen matrix. The ameba are then fixed, dehydrated and embedded within the Epon bullet. Early sections (the ones first off) are of interest since the ameba tend to attach to substrate.
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Biological Physics. Postdoctoral position at the University of Virginia. A postdoctoral position is available in the Department of Molecular Physiology and Biological Physics of the School of Medicine. The research projects in which the applicant is expected to play a major role involves electron energy loss spectroscopy and energy filtered scanning transmission electron microscopy. The position will include both development of software and instrumentation for achieving 2 to 3 nm spatial resolution compositional imaging and hands-on application of the method to significant biological problems. The laboratory has been engaged in NIH- supported research developing and applying analytical electron microscopy for 25 years through an interdisciplinary program based on the collaboration between physicists and biologists. Equipment available includes a 200kV electron microscope equipped with field emission gun (Philips CM200-FEG), a GATAN electron spectrometer adapted to our own CCD camera, a CM12 electron microscope and two energy dispersive X-ray detectors. Investigators in the program are also members of the Center for Structural Biology of the University of Virginia and have programs involving collaborations with the X-ray crystallography and atomic force microscopy groups and with molecular biologists and investigators engaged in other biological disciplines.
Candidates should have a Ph.D. in Physics, material science or engineering and be familiar with computer programming, instrumentation and, preferably, experience with electron microscopy and electron energy loss spectroscopy. Applications with biographical sketch, bibliography and the names of three references should be sent to: Dr. Andrew P. Somlyo, Department of Molecular Physiology and Biological Physics, University of Virginia, P.O. Box 10011, Charlottesville, VA 22906-0011, USA. The University of Virginia is an Equal Opportunity/Affirmative Action Employer.
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} Date: Mon, 9 Jun 1997 09:54:07 -0500 } To: HILDEGARD CROWLEY {hcrowley-at-du.edu} } From: Tom Phillips {tphillips-at-biosci.mbp.missouri.edu} } Subject: Re: TEM:Help!Correct PBS formula???? } Cc: } Bcc: } X-Attachments: } "Saline" is a physiological term for 0.9% NaCl solution which is 0.154 M. This is osmotically correct for human serum. If you add a large amount of phosphate buffer (e.g., 0.1M), one would have to lower the NaCl concentration in an equivalent manner if you want to maintain osmolarity. The Gibco catalog gives two formulations for PBS on page 2.9 of their 1997 catalog. one lists 9.0 g NaCl, 0.795 g NaH2POH-7H2O and 0.144 g KH2PO4 (all per liter). this is about 3 mM Na2PO4 and 1 mM KH2PO4. the other formulation is 0.21 g KH2PO4 and 0.726 g NaH2PO4-7H20. either one causes a slight hyperosmolarity in respect to straight "saline". In osmicated tissue, osmolarity might not be important but I suspect in aldehyde fixed tissue it can be. In TEM, one has to worry about PO4 based buffers causing precipitation of Ca salts. In procedures like DAB, one has to ensure the amount of buffer capacity is not exceeded. } } } All over our building we have an argument raging as to the correct } } formulation for PBS. Mainly the argument consists of opinions that the } } phosphate concentration should be 0.1M, and other opinions that the } } concentration ought to be O.OlM. (We all agree that NaCl concentration } } is 0.15M). There are also conflicting statements in the literature as to } } the composition of PBS. } } What we need is a thorough explanation of why one or the other } } concentration of phosphate salts is correct for mammalian tissue for TEM } } in procedures such as the DAB process. } } I would so much appreciate help in logically settling this argument around } } here. } } So long, } } Hildy } } } } Hildy Crowley } } University of Denver } } 2101 E. Wesley Ave } } Denver, CO 80208 }
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
Do any of you folks have any experience using the Polaroid MicroCam with a Nikon Labophot-2 or equivalent microscope with Color-Free objectives? The MicroCam replaces the eyepiece of the microscope; it is unclear to me whether or not the MicroCam includes compensation for lateral chromatic aberation (also known as chromatic difference of magnification). The Nikon (as well as other recent microscope) objectives are corrected to provide a color free intermediate image. If the MicroCam includes compensation, it seems to me that this might negatively affect the resulting photomicrographs.
I have been unable to obtain any information from Polaroid telephone technical support on this question.
Does anyone have any experience with using a Pixera digital camera for routine light microscopy work? If so could you share your experiences/feelings/comments either to the list or off-line?
What I'm looking for:
(1) a means of directly capturing digital images from routine light microscopy (i.e. bright field, phase etc.) but not barely even visable fluorescence (i.e. no FISH!).
(2) Speed is not a factor, as we aren't interested in highend "live" video, just static images, and scan times of up 1 minute would not be a problem.
(3) Resolution is a factor: i.e. really would like better than tv resolution (i.e. 640x480), beter than 1024x1024 would be perferable.
(4) Must be color.
(5) PC systems only (i.e. Pentium Pro/PCI/Windows NT), but PCI, or SCSI is o.k..
(6) And of course $$ is always a factor (yes, I'd love a microlumina, but I only have ~$1,500 to spend on camera).
The pixera seemed like a really good compromise on resolution and price (ah, but theory and reality are two different things).
Thank you in advanced.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513-529-5712 Fax: 513-529-4243 E-mail: edelmare-at-muohio.edu
"WE ARE MICROSOFT. RESISTANCE IS FUTILE. YOU WILL BE ASSIMILATED."
I would like to announce the following open position; which is sepearte from and in addition to the one I posted for a senior analyst/microscopist on 4/23/97. Please spread the word-Thanks
A position is currently open for an analyst at the North Carolina State University Analytical Instrumentation Facility (AIF).
Duties and responsibilities include day to day operation and maintenance of a Cameca IMS-6F SIMS instrument, stylus profilometers, and sample preparation devices such as plasma metal coaters, vacuum ovens, etc; assistance with scheduling of access to and oversight of the above instrumentation, assistance with the teaching of surface analytical laboratory classes and assistance with other graduate level engineering classes. Requirements include a BS or higher degree in analytical chemistry or a materials related discipline (non biological) along with some hands on analytical experience, equivalent to the MS or PhD level. Experience in a multi-user SIMS facility highly desireable. Experience in vacuum equipment and/or electronics maintenance; experience with PC and Unix; and experience with analysis of semiconductor or other non biological materials a plus.
Please send resume, salary history and names of references to: Prof. Phil Russell,Director Analytical Instrumentation Facility; North Carolina State University; Box 7531, Room 118A EGRC; 1020 Main Campus Drive; Raleigh, NC 27695-7531
Email application is encouraged to prussell-at-ncsu.edu. Fax 919 515 6965
North Carolina State University is an Equal Opportunity, Affirmative Action Educator and Employer. Minority and Female Applicants are especially encouraged.
Phillip E. Russell Analytical Instrumentation Facility Box 7531, Room 318 EGRC North Carolina State University Raleigh, NC 27695-7531
In my experience the Polaroid MicroCam has several drawbacks that render it useless, at least for my applications, low and meduim mag of stained mammalian histological specimens.
1. Resolution is poor on the film no matter how carefully one focuses througth the camera.
2. Color reproduction of typical biological dyes (hematoxylin, eosin, analine blue, orange G, PAS, etc) is very poor.
So much for your concerns about chromatic abberation!
3. The camera only takes (expensive) Polaroid print film so you won't have a negative to print or a slide to project.
4. Exposue varies with the specimen so many test shots are necessary.
The results I saw at two different demos were so poor that if I had a free MicroCam and a free lifetime supply of Polaroid film, I still would not use it! I was amazed that Polaroid would try to sell such a piece of junk!
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 e-mail: mcauliff-at-umdnj.edu ***************************************************************
A very useful paper to help choose the best buffer concentration and how much of common additives is: MD Maser et al (1967), Relationships among pH, osmolality and concentration of fixative solutions, Stain Technology 42:175-182. It also describes why it is important to check the osmolality and has a useful list of further reading. Diana.
Diana van Driel Dept Ophthalmology Sydney University C09 AUSTRALIA 2006
- Antique or scientific instrument dealers as well as Sotheby's or Christie's or any other local auction house should be able to value such a rarity.
- The microscope journal "MIKROKOSMOS" Stuttgart (Gustav Fisher Verlag) also publishes "microscopes for sale".
- A brass microscope in excellent condition of LEITZ or ZEISS from early this century could cost about 1500 to 2000 DEM, or $900 to $1500 US, depending on the condition and sorts of accessoires.
- Microscopes from individual manufacturers are in general lower, but estimated on a more subjective basis between nothing, 250$ and very much more than 2000 DEM. If a microscope is not from a famous manufacturer, optics are generally estimated as comparably poor, which in the case of this microscope is more or less the case: it is not possible to focus more than about 60% of the field of view at the time.
Some were concerned why I would sell such a device which was manufactured by an ancestor. I did not want to upset anyone. So if you have time and if you are interested:
My great-grandfather was a very passionate precision mechanic with an unusual high professional pride. That must be why he passed his spare time building tons (to give you an idea, the approximate cost of only having the completely unusable part of his tools put away two weeks ago were about 1000 DM) of devices like a precision scale, microscopes, a telescope, carved wooden statues, building furniture, as well as countless oil paintings etc. etc. Also, he was known to having sold *his* ancestors things to be more mobile, as he was not the first one in the family to build things together.
Rather, we seem to be a whole tribe of such persons dedicated to the combination of anything with manual work. My grandfather (mechanic, passionate builder of exquisite wooden furniture still in use throughout the family, who carved some of the family into statues), my mother (certified master of graphoanalysis I.G.A.S., passionate manual worker with beautiful oil and water color paintings to entirely fill our house, wood, cloth etc.), and her sister (a physicist who worked in the field of semiconductor crystal epitaxy and earned an IBM research prize for that, passionate piano player), and her brother (electrician, passionate oil painter), and me (on my way to forensic pathology, passionate oil painter, wood carver, playing guitar, at high-school I was doing technical and comic drawings as well as giving guitar lessons aside to earn some money; in my first clinical year, I stitched together among others a complicated chainsaw skin lesion of about 2 x 4 cm using about 50 several stitches, which healed without complications) and my sister (physical therapist, working more with cloth and clay: you just should see her flat!), as well as our cousins (one engineer and draughtsman and passionate violinist who just played in a cathedral at Einsiedeln here in Switzerland, one also physicist doing semiconductor technology, two others medical doctors) all must have some of those genes: we are all passionate about manually working in our most interesting fields, we all virtually have the inability to do other in our spare time than to carve, paint, play instruments etc. and each amass our specific load of manufactured products, each in his particular area. For the family of my father: My father is working at the Robotics department here, his brother is an architect and passionate painter as well. Other relatives: my other great-uncle was a Saddler/upholsterer who built his small store into a Prime furniture house, and the one great-uncle who died at the age of 102 incredibly enjoyed playing piano until he was at least a century old, his daughter was a professional seamstress with her own business. But we all have also the strong wish to give at least some of our ancestors devices away for more mobility.
Also I just realize that we are continuing this tradition: My girl friend (physical therapist with back-breakingly stuffed schedule despite rising health cost, passionate wood carver as well who finished her first cat statue last summer) and my sister's husband (surveyor and passionate mechanic) also love their heavily manually oriented work, as well as their relatives do (examples: one is a mechanic producing technical solutions for his firm, then, one is a medical doctor in the field of orthopedics).
Because I have been asked by some distant relative, who would very much love to buy and exhibit one of his microscopes in her living room, we are probably going to offer this item as a gift or for a reasonably low price. Surely we would not dishonor our ancestor in any way !
As our family history goes back to about 1640, I am very grateful they were such great people about their work and hobbies and everything, as am I and all around here, but I am actually quite glad they did not keep *all* those things they all happily put together primarily out of the satisfaction just to do it, and I am especially glad they are doing Quality Control over there at LEITZ's and ZEISS'.
And I especially acknowledge your help and understanding, Wolf Schweitzer
I am an Information Research Specialist with Motorola SSTG in Scottsdale, Arizona. I am having difficulty locating a certain document and thought this list might be of assistance in my search. Any help or suggestion is greatly appreciated.
The document I am looking for is :
"Auger, EMPA, XPS, and EBSP Analysis of Discolored Au/Ti/Cu Multilayer Thin Film I/O Pads", R. E. Davis, J. L. Hurd, J. Gates Jinkins, and R. Flitsch, Microbeam Analytical Society, Aug. 1993, Los Angeles, CA
If you know of this document and where I may obtain it, please contact me via email directly at p28167-at-email.mot.com.
Thank you in advance for your assistance and best regards,
Donald Guy
Information Research Specialist SSTG Learning Center/Library Motorola Space and Systems Technology Group 8220 E. Roosevelt St., MD R1206 Scottsdale, Arizona 85252
We have been using a Web-based sign-up system for our confocal for about a year and like it very much. The system can be seen via the "Confocal Schedule" link on the following Web-page:
http://ww2.med.jhu.edu/confocal/
Only registered (i.e. properly trained) users can sign up, using their password, up to one month in advance. The system is such that any person's time can be cancelled by any other user in their own lab -- very handy in the case of illness, experiments going longer than expected, or the boss informing you that you really should be working on project X rather than Y. The facility does charge a "no-show" fee for folks who reserve time and don't use it. This is a putative measure that can always be avoided (even for last minute changes in plans) by informing the person in charge of billing and the person signed up after you.
Greg Martin Dept. of Cell Biology and Anatomy Johns Hopkins School of Medicine
On Wed, 4 Jun 1997, Tamara Howard wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Does anyone have on-line sign-up for equipment time? I remember this was } discussed here ages ago, but I haven't heard about it recently. We are } thinking about going to such a system, and would like to hear } pros/cons/horror stories/suggestions. } } Thanks! } } Tamara } CSHL (NY) } howard-at-cshl.org } } }
Hello all, What is the best way to process cells for TEM which are grown on glass coverslips affixed to plastic petri dishes. I will need to remove the glass from the plastic and cut ultra thin sections. I have previously tried to do this and could not remove the glass. I gave up on glass and started using aclar. I now must return to using glass. Any suggestions?
Sometimes if all else fails, I take a wood stick, dip it in to chloroform and wave it over the sections in the boat like a magic wand. If its not an infiltration problem, this can flatten the sections.
Bob U of Washington Seattle
On Mon, 9 Jun 1997, William Meek, Ph.D. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I am having problems with wrinkles in Epon thin sections (gold } interference colors)and would like some advice or information about } getting rid of them. The specimen is ameba and has been grown on the } surface of an Epon bullet coated with a collagen matrix. The ameba are } then fixed, dehydrated and embedded within the Epon bullet. Early } sections (the ones first off) are of interest since the ameba tend to } attach to substrate. } } Bill }
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We will be receiving an FTIR with a microscope accessory soon. I would appreciate comments about this technique from users whose application is drug identification or fragment ID of particulates sometimes found on food. Does it truly have the power to zero in on the suspect substance without sample extraction/cleanup? Formerly we used diffuse reflectance or made pellets for FTIR spectra of our old spec without a microscope. Thanks chemist SLI
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sally: Our EM facility has been using glass coverslips in several tissue culture experiments. After flat embedding with epoxy/alradite resin, we remove the glass, using cold HF[hydroflouric acid] (in a Fume Hood!) for -at-4-8 minutes. We get 100% success rate, using many different types of glass coverslips. However, the glass cover slips are not attached to plastic petri dishes. Do the coverslips need to be attached to petri dishes? The glass surface has to be exposed to the HF fluid. You can still use this technigue if your coverslips are atteched. you will have to add an extra step to remove the glass from the plastic, via sandpaper. Please contact me, if you would like a detailed step by step procedure for this technique. It is very straightforward.
The references we started with: Moore, M.J., 1975, Removal of Glass Coverslips from Cultures Flat Embedded in Epoxy Resins Using HF, . Microscopy, v. 104, p. 205.
Rieder, Conly and Bowser, Samuel, 1987, "Correlative LM and EM on the Same Epoxy Section", in Correlative Microscopy in Biology , Chapter 11, Academic Press, 1987.
-Louisa howard Dartmouth College EM Facility Hanover, NH. 03755 (603) 646-3492 Louisa.Howard-at- Dartmouth.edu
We are using some classical light microscopy staining techniques to screen our plant samples before we carry out EM on them. Many of these techniques include carrying the samples through xylene baths. We have been encouraged to find a safer alternative, if possible. I remember that years ago a xylene replacement was introduced into the marketplace to be used during paraffin embedding of samples, but I heard that this was not a good replacement, and it was discontinued. I don't know what happened after that or what is used these days.
Any help on this matter would be greatly appreciated. Thanks.
Paula.
Paula Allan-Wojtas Food Microstructure Specialist Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4R 1A1
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello all, } What is the best way to process cells for TEM which are grown on glass } coverslips affixed to plastic petri dishes. I will need to remove the } glass from the plastic and cut ultra thin sections. I have previously } tried to do this and could not remove the glass. I gave up on glass and } started using aclar. I now must return to using glass. Any suggestions? } } Sally } }
Group A reader of mine, not on this listserver, is "interested in obtaining electron micrographs of a number of microbes: T. Pallidium, S. pneumoniae, H. pylori, V. Cholerae, m. tuberculosis, and others." Should you be able to help, please contact Bruce Cameron directly at: bcameron-at-tigr.org Regards to all, Don Grimes, Microscopy Today
The newest version of the Image Processing Tool Kit is now completed. The CDs are being sent free of charge to upgrade all registered users of the kit. Version 2.1 includes about 80 Photoshop compatible plug ins with image processing and measurement functions. Many of the plugins offer speed improvements of 2x to more than 10x over version 1, and are fully compatible with the Layers structure used in Photoshop 4. The Mac and PC versions (both on the CD) are the same. The CD includes a much expanded tutorial (175 pages) and more than 100 test images. It also has versions of the tutorial that specifically discuss the use of the tool kit with NIH-Image (Mac) and ImageTool (Win). Full info on the package is available from http://members.aol.com/ImagProcTK/ A review of the original CD is available on-line from http://www.macscitech.org/stj/stj1997_apr/stj1997_apr.html#one
Jim, I tried immersing my plastic in liquid nitrogen and was unable to remove the glass. Perhaps you did something special during the whole process of embedding, or perhaps I am not using the best plastic. I use embed 812 kit. Also, I have heard that if you use serum in the culture medium, then the glass will not come off the surface of the plastic. Does this make sence?
Thanks for the info on Histoclear. I've been using a supply that was=20 here before me and I wasn't sure if it was available anymore or where to=20 get it. It's interesting to read that it's not for use with paramount- I= =20 use it all the time, even use the Histoclear to thin the paramount when=20 it starts to get too thick. I haven't noticed a big problem, although it= =20 takes the coverslip alot longer to dry down. In one of my applications,=20 I rinse celloidin sections in Histoclear prior to pressing them flat on a= =20 slide and trimming the edges of the section with a razor blade. I like=20 Histoclear better for this because is does take a while to evaporate and=20 gives me more time to work with the sections before they dry out. (They=20 need to stay moist throughout the mounting process.) =20
One more note. Althought Histoclear is supposed to be nontoxic, it gives= =20 me a headache, so I always work in a hood.
Karen
On Tue, 10 Jun 1997, Philip Oshel wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } =20 } } We are using some classical light microscopy staining techniques to } } screen our plant samples before we carry out EM on them. Many of these } } techniques include carrying the samples through xylene baths. We have } } been encouraged to find a safer alternative, if possible. I remember tha= t } } years ago a xylene replacement was introduced into the marketplace to } } be used during paraffin embedding of samples, but I heard that this was } } not a good replacement, and it was discontinued. I don't know what } } happened after that or what is used these days. } } } } Any help on this matter would be greatly appreciated. Thanks. } } Paula Allan-Wojtas } =20 } Paula, } =20 } There is a fine replacement available (unless it's been taken off the } market in the last year): HistoClear, marketed by National Diagnostics. } This is also sold by Fisher, or if they don't have HistoClear, they have } their own brand of the same thing. } =20 } It's "essential oils of citrus", and smells like oranges. Non-toxic, work= s } very well for paraffin sectioning, or any other procedure that uses } xylenes. But (there's always a but), you can't use Permount or the like t= o } mount the coverslips. N. D. has a replacement product HistoMount (as does } Fisher, I believe). This works very nicely, But ... , you can't warm the } slides to set the mountant as you do with Permount, etc. Otherwise the } HistoMount shrinks like mad. Shrinks some anyway, but this in only a majo= r } annoyance if you're doing very thick sections. The usual 10-15=B5m ones a= re } no trouble. } =20 } Note: companies are mentioned because I think National Diagnostics } developed (or first commercialized) this stuff, and because I know they a= nd } Fisher sell it. Other companies probably do as well. (N.D. also came out } with a replacement for foraldehyde as a fixative, using a sugar aldehyde. } This didn't seem to work so well, and *maybe* has been withdrawn, but I } don't know.) } =20 } Phil } =20 } } Sic Hoc Legere Scis Nimium Eruditionis Habes { } Philip Oshel } Station A } PO Box 5037 } Champaign, IL 61825-5037 } (217) 355-1143 } oshel-at-ux1.cso.uiuc.edu } *** looking for a job again ****************** } =20 } =20 } =20 } =20
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Subject: Time:7:32 AM OFFICE MEMO Post-doc position at LLNL Date:6/11/97
Our Center for Accelerator Mass Spectrometry (CAMS) is currently seeking a Postdoctoral Research Staff Member to work as part of an interdisciplinary team of physicists, chemists, and biologists. In this research position, you will work as part of a team to develop quantitative elemental analysis of biological tissues. These techniques will be used in studies of elemental kinetics, structural biology, toxicology or pharmacology by staff at LLNL and collaborators in the University of California system and other institutions. Duties include development/modification of techniques for preparation of biological tissues for quantitative elemental microanalysis, operation of the LLNL microprobes to analyze prepared biological samples, X#030#ray data reduction and analysis, planning/designing and executing independent and collaborative research projects and publication of results in peer#030#reviewed literature. Requires a recent Ph.D. in chemistry, biochemistry, toxicology, pharmacology or related field. Experience in trace element analysis and analytical microscopy is desired. LLNL offers a challenging work environment and a competitive salary/benefits package. Qualified individuals are invited to send their resume to: Lawrence Livermore National Laboratory, Attn.: Mary Anne Holman, L#030#725, P.O. Box 5510, R#030#4249, Dept. AJSC527MH, Livermore, CA 94551#030#5510. LLNL is an Equal Opportunity Employer. Lawrence Livermore National Laboratory
I have been using micro-FTIR for years to identify particle and fiber contaminants in pharmaceutical products. Your statement about being able to identify particulates without extraction is partially true; it depends alot on the matrix in which the contaminant is found. I have found it useful to at least "rinse" the contaminant in a solvent (in these applications the solvent is usually water) first. Otherwise, you may have trouble with minor extraneous absorptions which hamper the positive identification of your contaminant, especially with automated search routines.
I have found that a low power (10-60x) stereomicroscope with dark field lighting is imperative in the pre-analyis prep of such samples. A clear glass spot plate (or microscope slide with wells) can be used for extraction/cleanup of your particle. You can view it under the 'scope while the extraction occurs. You also should have a good supply of micro-probing tools (available from most SEM supply houses) if you don't already have them. If you "tease" the particle around in the solvent, and pull it away from the solvent as it evaporates under the heat of the 'scope you can usually clean up the particle enough to obtain a good spectra.
Particle size is another consideration. Typically, you can obtain a useful transmission spectrum from particles greater than about 20 microns. Reflectance spectra may require larger sizes. Fibers generally work quite well in transmission. In fact, I once built a library of spectra for the types of fibers found in the cleanroom garments used in the compounding area for a pharmaceutical product. I subsequently used that library to identify/troubleshoot particulate problems for the customer.
I hope this info is useful to you. Good luck in your identification work.
Regards,
Bob ***************************** Bob Citron Chiron Vision Corp. Claremont, CA 91711 USA (909)399-1311 Bob_Citron-at-cc.chiron.com *****************************
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We will be receiving an FTIR with a microscope accessory soon. I would appreciate comments about this technique from users whose application is drug identification or fragment ID of particulates sometimes found on food. Does it truly have the power to zero in on the suspect substance without sample extraction/cleanup? Formerly we used diffuse reflectance or made pellets for FTIR spectra of our old spec without a microscope. Thanks chemist SLI
X-Authentication-Warning: duchesse.zdv.Uni-Mainz.DE: windoff owned process doing -bs
Hello, I am working with cells growing on round cover slides. We are looking for an apporbiate holder to handle them in larger amounts during immunohistochemistry. Something like a smaller version of the staining jars which are used for normal microscopic slides. Can someone help me?
I use a very nice ceramic coverslip holder purchased from Thomas Scientific. You can also purchase glass jars of the appropriate size from the same catalog. Note that this item may be available from other vendors as well. This holder carries a dozen coverslips and has a metal handle that you can remove. It's actually a miniature version of the regular slide holders. I used it with square coverslips but regular sized round covers should hold without problem. Sorry, I don't have the current reference at hand. Good luck, Michel
**************************************************** Michel Deschuyteneer, Ph.D. deschuyt-at-sbbio.be Scientist Electron Microscopy Laboratory
SmithKline Beecham Biologicals Rue de l'Institut, 89 B1330 Rixensart, BELGIUM Tel: +32-2-656 9290 Fax: +32-2-656 8164 **************************************************** Standard disclaimer: the opinions expressed in this communication are my own and do not necessarily reflect those of SmithKline Beecham. ****************************************************
} Hello all, } What is the best way to process cells for TEM which are grown on glass } coverslips affixed to plastic petri dishes. I will need to remove the } glass from the plastic and cut ultra thin sections. I have previously } tried to do this and could not remove the glass. I gave up on glass and } started using aclar. I now must return to using glass. Any suggestions? } } Sally
Have you tried using Thermanox coverslips? Not cheap, but they are good for culturing and good for embedding and they peel away from resin with the application of a little heat from a hot-plate. Thermanox can also be sectioned along with the resin if you like, but I have had problems with this because the Thermanox and resin do not stretch to the same degree, so you end up with sections wrinkled on one edge.
We have "popped" cells grown in both serum and serum-free medi on glass coverslips using liquid nitrogen 100's of times. serum makes no difference. The coverslips are fixed either in the 12 well plastic trays or transfered to 20 ml glass scint vials. by the time we get to the dehydration steps, we always switch to the glass vials. after 100% ethanol, we infiltrate with embed 812 (1:1 with ethanol overnight) then 100% resin for 3 hrs then place the coverslip cell side up on a glass slide with the thin layer of resin that comes with it when we transfer to the slide (you don't want it too thin or too thick but it if you are in doubt, simply add 1 drop to the coverslip after transfering and let it flow naturally. you shouldn't get a final plastic spread of more than 1.5 - 2 times the diameter of the coverslip or you are using too much. when you take the slides out of the oven, let them cool for 15 min and touch with a razor blade. if you get a gooey strand when you pull the blade away, you haven't polymerized it enough. if it is not too gooey, cross-hatch the surface of the plastic using the razor blade to score deeply into the plastic (to the level of the glass). SLOWLY immerse the slide into Liquid N2 and the squares should pop right off. look at the surface of the coverslip and bottom of the square to ensure no glass is going with the square. if so, you have over polymerized. for some projects, we simply put the square in the flat microtome holder and cut it but ususally we re-embed the square in a standard mold. if you aren't re-embedding, you should heat the squares in a rubber mold overnight. once you have the timing of how long to polymerize the coverslips, it is trivial. we havent had a failure in years of doing this a lot of times. good luck.
Tom Phillips
} Jim, } I tried immersing my plastic in liquid nitrogen and was unable to remove } the glass. Perhaps you did something special during the whole process of } embedding, or perhaps I am not using the best plastic. I use embed 812 } kit. Also, I have heard that if you use serum in the culture medium, then } the glass will not come off the surface of the plastic. Does this make } sence? } } Sally
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
This fall and winter I need to fix and embed dormant blueberry buds; it seems likely that I will run into problems trying to infiltrate the somewhat woody exterior with fixatives and the embedding medium. We're going to be immunolabeling so we'll be using LR White but I'm wondering if anyone with experience with woody tissue can recommend buffers and fixatives ?
We generally use 0.2% glutaraldehyde, 3% paraformaldehyde in a 0.05M buffer and this works fine for non-woody tissues so I thought I'd start there but I would greatly appreciate any advice!
} there that can do this faster. Does anyone know of such a software? I } would appreciate comments.
GLOBAL LAB (Data Translation)
/-------------------------------------------------------------------------\ | Rui Costa | e-mail: ruicosta-at-esb.ucp.pt | | Escola Superior de Biotecnologia |telephone: 351-2-5580044 | | Rua Dr Antonio Bernardino de Almeida |fax: 351-2-590351 | | 4200 PORTO | | | PORTUGAL | | \-------------------------------------------------------------------------/
I've been asked to help some colleagues monitor the production of hydrogen peroxide in potato leaves undergoing bacterial attack. I have a paper citing a technique using CeCl3 - cut samples are incubated in the cerium prior to fixation (glutaraldehyde, paraformaldehyde, sodium cacodylate buffer , post-fixed in osmium), dehydration (ETOH and propylene oxide) and embedding (Eponaraldite). I've never done this before and am wondering: can anyone give me some helpful tips, hints, advice etc?
Hi, folks. I've inherited an Orthomat camera (old-style, not the more recent Wild-Leitz type) without the film cassette. If you have one or more that you'd be willing to part with for $$ or barter, please contact me at smithj-at-winthrop.edu. TIA Julian
Julian P.S. Smith III Biology Winthrop University Rock Hill, SC 29733 803-323-2111 x227 (vox) 803-323-2246 (fax)
I have come across the use of 'Histoclear' - sorry that's all I know about it. But in recent years we have used an even safer alternative called 'Citroclear' (I believe its an extract of citrus fruits) - in the UK this is supplied by: HD Supplies 44 Rabans Close Rabans Lane Industrial Estate Aylesbury Bucks HP19 3RS UK Phone Aylesbury (01296) 431920 - I will leave you to work out the international dialling codes Fax Aylesbury (01296) 392121
The cost in our 1995 price list was: catalogue number - CC500 Citroclear 5litres - 16.75 UK pounds 25litres - 74 UK pounds the prices are ex VAT(purchase tax) ex delivery and probably out of date.
It appears to do much the same job as xylene but is ( at this present time) considered to be a lot less toxic. It's hazard data sheet lists it's known hazards as an irritant with the possibility of dermatitis after long exposure. Other characteristics are a strong fruity smell which some people like and some don't and a tendency to turn yellow and throw out oily deposits with prolonged storage in light. Oh the suppliers say it is bio-degradable as well and can be carefully disposed of down the drains if your regulations allow.
I don't know if you can source this in Canada but good luck in your search. By the way I have never used xylene outside of a fumehood in years.
Malcolm Haswell University of Sunderland UK
Disclaimer - I have no connection with the company other than as a satisfied user.
----------
Separating glass from plastic is easy in my experience after thermal shock. Immerse the specimen in liquid nitrogen for a few seconds. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 350+ Links, MSDS ************************ http://www.proscitech.com.au
} From: Sally Shrom {sally-at-retina.anatomy.upenn.edu} } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM cell culture } Date: Tuesday, 10 June 1997 23:13
} } Hello all, } What is the best way to process cells for TEM which are grown on glass } coverslips affixed to plastic petri dishes. I will need to remove the } glass from the plastic and cut ultra thin sections. I have previously } tried to do this and could not remove the glass. I gave up on glass and } started using aclar. I now must return to using glass. Any suggestions? } } Sally
If the usual stretching with solvent or heat (which is required to compensate the compression artifact) does not do the job, try using a loop to pick up the section. Then deposit these onto a grid which is sitting on filter paper. This solved a wrinkle problem I had with thick cell walls. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 350+ Links, MSDS ************************ http://www.proscitech.com.au
} On Mon, 9 Jun 1997, William Meek, Ph.D. wrote: } I am having problems with wrinkles in Epon thin sections (gold } interference colors)and would like some advice or information about } getting rid of them. The specimen is ameba and has been grown on the } surface of an Epon bullet coated with a collagen matrix. The ameba are } } then fixed, dehydrated and embedded within the Epon bullet. Early } sections (the ones first off) are of interest since the ameba tend to } attach to substrate. } } Bill
} We are using some classical light microscopy staining techniques to } screen our plant samples before we carry out EM on them. Many of these } techniques include carrying the samples through xylene baths. We have } been encouraged to find a safer alternative, if possible. I remember that } years ago a xylene replacement was introduced into the marketplace to } be used during paraffin embedding of samples, but I heard that this was } not a good replacement, and it was discontinued. I don't know what } happened after that or what is used these days. } } Any help on this matter would be greatly appreciated. Thanks. } Paula Allan-Wojtas
Paula,
There is a fine replacement available (unless it's been taken off the market in the last year): HistoClear, marketed by National Diagnostics. This is also sold by Fisher, or if they don't have HistoClear, they have their own brand of the same thing.
It's "essential oils of citrus", and smells like oranges. Non-toxic, works very well for paraffin sectioning, or any other procedure that uses xylenes. But (there's always a but), you can't use Permount or the like to mount the coverslips. N. D. has a replacement product HistoMount (as does =46isher, I believe). This works very nicely, But ... , you can't warm the slides to set the mountant as you do with Permount, etc. Otherwise the HistoMount shrinks like mad. Shrinks some anyway, but this in only a major annoyance if you're doing very thick sections. The usual 10-15=B5m ones are no trouble.
Note: companies are mentioned because I think National Diagnostics developed (or first commercialized) this stuff, and because I know they and =46isher sell it. Other companies probably do as well. (N.D. also came out with a replacement for foraldehyde as a fixative, using a sugar aldehyde. This didn't seem to work so well, and *maybe* has been withdrawn, but I don't know.)
Phil
} Sic Hoc Legere Scis Nimium Eruditionis Habes { Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217) 355-1143 oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
} } I am having problems with wrinkles in Epon thin sections (gold } } interference colors)and would like some advice or information about } } getting rid of them. The specimen is ameba and has been grown on the } } surface of an Epon bullet coated with a collagen matrix. The ameba are } } then fixed, dehydrated and embedded within the Epon bullet. Early } } sections (the ones first off) are of interest since the ameba tend to } } attach to substrate.
Usually the first sections to come off of organisms growing on surfaces tend to be somewhat problematic probably due to the the irregularities of the surface and the modifications of the cell that facilitate adherence to the surface. Such sections tend to wrinkle and may split apart in some cases. We find that picking up the sections onto a naked grid (flamed nickel grid, for example) and passing the sections (still floating on a droplet of water on the grid) between a heated electrical loop will tremendously relax the sections - more so than chloroform. If this does not work, then consider varying the clearance angle of the knife as well as the sectioning speed, increasing the hardness of the plastic, or use a good, smaller angled (or diamond) knife. Good luck.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 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 ####################################################################
Are your sections dried down onto a formvar or other support film? Wrinkles can also occur when sections "loosen" during the staining process and then subsequently dry down again (unevenly). This can be remedied by firmly attaching the sections to the support film by heating the grids at 60 C for 15-30 min. This is best done using a heating block. Good luck.
Tim Bourett DuPont Experimental Station Wilmington, DE USA
I am seeking to purchase used, the epi-fluoresence accessories for a Leitz Orthoplan that is over 20 years old.
Please reply privately.
Thanks, Greg Erdos ******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
We have used FTIR microscopy routinely in our labs for a few years and are enthusiasticly for it. Two of the most important advantages are the simple sample preparation and the ability to take spectra of small particles. We often wish to find polymorphs (which may be a small percentage of the whole) of investigational drug substances and FTIR allows us to look at individual particles. Its great for aiding in the identification of small bits of polymer contaminants which can be tough by polarized light microscopy.
One of the limitations is a familiar one to all of microscopy - sampling. How can one know whether the selected particle is truly representative of the whole? We also think it is necessary to crush the particle to reduce IR polarization effects. The minimum size given by some of the manufacturers also seems a bit small in normal applications. We like particles in the 30 to 70 um region.
Our routine sample preparation technique involves mounting the particle or particles on a KBr window under a stereomicroscope. We then gently crush the particle with a stainless steel roller. We then either mark the location or keep in mind the surroundings. One problem we've had is finding the particle of interest using the optics of the IR scope. We are then ready to test.
I have a big problem, osmium precipitates!! does anyone can help me? Thanks in advance Nuria Cortadellas Department of Electron Microscopy University of Barcelona
I'm sorry I can't address the question about the corrections of the Polariod Micro camera but I would like to respond to the torching it received. I agree that the results are far from critical photomicrography, but it does have a niche in my lab. I have a stereomicroscope without a camera and there are times that I require simple, quick photo documentation. For instance, I just worked on some tablets with unsightly spots and I wanted to document their location and color before I analyzed the spots by EDS. The Polaroid Micro worked quite nicely. The thing I like the most is that it is an SLR so I get to see exactly what I'm photographing and can judge focus easily. Its surely not a Cadillac and probably not a Chevy but it may be a Moped.
} Hello, } I am working with cells growing on round cover slides. We are looking for } an apporbiate holder to handle them in larger amounts during } immunohistochemistry. Something like a smaller version of the staining } jars which are used for normal microscopic slides. Can someone help me? } } Thanks } Reinhard } } . . . . . . . . . . . . . . . . . . . } Dr. Reinhard Windoffer Fon: (00)49 (0)6131/39 3720 } Universitaet Mainz Fax: (00)49 (0)6131/39 4615 } Anatomisches Institut e-mail: windoff-at-mail.uni-mainz.de } Becherweg 13 } D-55099 Mainz } Germany } . . . . . . . . . . . . . . . . . . . Reinhard, I have made round coverslip holders from fairly thick walled flexible polyethylene and silicone tubing by cutting it in half lengthwise and then cutting slits perpendicular to the length. When the tubing is flexed in the correct direction the slits are open for inserting the cover slips. When the tubing is held straight (in a pyrex tube or syringe barrel) the slits pinch closed and hold the cover slips. Use a tubing diameter near that of the cover slips. A cautionary note, if you plan to critical point dry samples held this way, be advised to test the tubing material in the CPD unit first. Some tubing types will turn into foam during CPD.
good luck
cheers
ed
Edward J. Basgall, PhD The Pennsylvania State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618
} } Hello, } I am working with cells growing on round cover slides. We are looking for } an apporbiate holder to handle them in larger amounts during } immunohistochemistry. Something like a smaller version of the staining } jars which are used for normal microscopic slides. Can someone help me? } } Thanks } Reinhard } } . . . . . . . . . . . . . . . . . . . } Dr. Reinhard Windoffer Fon: (00)49 (0)6131/39 3720 } Universitaet Mainz Fax: (00)49 (0)6131/39 4615 } Anatomisches Institut e-mail: windoff-at-mail.uni-mainz.de } Becherweg 13 } D-55099 Mainz } Germany } . . . . . . . . . . . . . . . . . . . Reinhard, In my previous response I forgot to mention that a company here in the US, Tousimis Research Corp. also sold, at one time, a metal holder designed for CPD of stacks of cover slips. I have used it with good results for CPD. They come in two sizes. 13mm and 25mm? Ph: 800-638-9558 or 301-881-2450. (No affiliation, only a satisfied customer)
cheers ed
Edward J. Basgall, PhD The Pennsylvania State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618
After reading the Glutaraldehyde/formaldehyde safety discussion, I've decided to add my note of warning. After working with GAL and formaldehyde and other chemicals of the TEM trade for 30 years with only slight reactions to them, I was exposed to a major formaldehyde spill. Not only did I suffer major respiratory tract irritation, but my lung collaped and I soon developed hypersensitivity to a wide range of volatile organic chemicals, including formaldehyde, GAL, liquid epoxy resins, solvents, 2-mercaptoethanol, pesticides (including herbicides), auto exhaust, etc. When exposed to these I get headaches and "fuzzy" in the head almost instantly, and a delayed hypersensitivity reaction in my pleural cavity about 30' to 2 hr later that is extremely painful. This lead to further pneumothoraces until I had lung surgury. Our hypothesis is that I have developed an immune response to formaldehyde-altered proteins of my lung or pleural cavity, as well as the limbic system (neural) response often seen in others with chemical hypersensitivity.
Needless to say, I recommend keeping your exposures to a minimum. All use of aldehyde fixatives should be in a well-functioning fume hood. Anatomy classes should have individual fume hoods for each dissection station and/or switch to non-aldehyde fixed cats for example (we did both). Incidently, my unofficial survey of former occupations of persons with MCS indicates that histology technicians have the highest incidence and some formal surveys show similar results.
It's definitely less fun trying to function (and very difficult to do research) in today's chemically-dependent society when you are hypersensitive to nearly every volatile organic chemical around. -BE CAREFUL WITH THE GAL AND FORMALDEHYDE!-
-Dennis
Dr. M. Dennis Goode Phone (301) 405-6917 Department of Zoology Fax (301) 314-9358 University of Maryland e-mail goode-at-zool.umd.edu College Park MD 20742 ************************************************************* "If the Lord Almighty had consulted me before embarking upon the creation, I should have recommended something simpler." -Alphonso X of Castile, 15th Century
I am trying to obtain the contour of wheat to build the three dimension model of wheat. The contour image of wheat is not good. It is not continuous. If someone has good idea or method, please let me know. I sliced the wheat using ultramicrotome with glass knife. Then I obtained the image of wheat slice. I tried to obtain the contour image using Imaging software. Thanks.
Question #1: I have a Haskris RO75 water chiller and I am in need of another; I have found ALL of the components to make it a double pumper though. Has anyone turned their single pump RO75 into a double pumper? What are the pitfalls? Am I nuts or just cheap?
Question #2: A client of mine wants to see if she can see the difference between elastin and collagin at the TEM level {she is looking for the relative amounts in various samples}. Is this possible? What are the references/protocols? Am I nuts to try this?
Thanks, John Grazul Rutgers University Electron Imaging Facility
Global Lab is no longer sold by DT. You can try Visilog from Noesis Vision Inc. We have a version available for 2,995.00$ US.
At 05:43 PM 6/11/97 +0100, Rui Costa wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
---------------------------------------------------------------------------- ------------- Luc Nocente Tel: 514 345 1400 Noesis Vision Inc. Fax: 514 345 1575 e-mail: ln-at-noesisvision.com http://www.noesisvision.com 6800 Cote de Liesse, Suite 200 St-Laurent, PQ H4T 2A7,Canada ---------------------------------------------------------------------------- -------------
I recently asked for information regarding a Zeiss SEM 6. Subsequent direct conversations with the donor clarified that, in reality, a free Zeiss 9A TEM is available from here in San Diego. It was donated to the school district and they have been awaiting funds to install it. Those funds never materialized, so the district now wants to pass it along. I have no personal knowledge of the working condition of the scope--I do know it is currently sitting in a warehouse. Anyone interested in getting this donation should contact Debbie at the Grossmont School District Instructional Services Dept. at (619)579-9711. They are going to dispose of it in the very near future.
Dr. Steven Barlow EM Facility/Biology Department 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/EM_Facility
} Hello plant pro's, } } This fall and winter I need to fix and embed dormant blueberry buds; it } seems likely that I will run into problems trying to infiltrate the } somewhat woody exterior with fixatives and the embedding medium. We're } going to be immunolabeling so we'll be using LR White but I'm wondering if } anyone with experience with woody tissue can recommend buffers and } fixatives ? } } We generally use 0.2% glutaraldehyde, 3% paraformaldehyde in a 0.05M } buffer and this works fine for non-woody tissues so I thought I'd start } there but I would greatly appreciate any advice! } } Thanks,
Peggy,
I am no plant pro, however I have had some experience with doing some plant stuff for TEM. I had a couple of users looking at grass samples and also some clover root nodules, As you would probably know, very hard to fix and infiltrate adequately. We used Microwave fixation for this purpose and also used the microwave to aid in the infiltration of the resin. Initially, this was not for Immuno but the results were far superior and in a much shorter time. We did fix some grass bits for immuno using the MW, but still had a few problems with infiltration at low temps (Lowicryl K11M). I would say that using the MW would be able to help you infiltrate your 'woody tissues' with LR White too.
Rich.
----------------------------------------------------------------------- Richard Lander Senior Technician South Campus Electron Microscope Unit c/- Pathology Department Otago Medical School P.O. Box 913 Dunedin New Zealand. Tel. National 03 479 7301 Fax. National 03 479 7254
"Southernmost EM Unit in the World!" ------------------------------------------------------------------------
I will be involved with operating an ESEM in the near future and need information about any courses available concerning this technology. Any information will be greatly appreciated.
I will be involved with operating an ESEM in the near future and need information about any courses available concerning this technology. Any information will be greatly appreciated.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Subject: Time:7:32 AM OFFICE MEMO Post-doc position at LLNL Date:6/11/97
Our Center for Accelerator Mass Spectrometry (CAMS) is currently seeking a Postdoctoral Research Staff Member to work as part of an interdisciplinary team of physicists, chemists, and biologists. In this research position, you will work as part of a team to develop quantitative elemental analysis of biological tissues. These techniques will be used in studies of elemental kinetics, structural biology, toxicology or pharmacology by staff at LLNL and collaborators in the University of California system and other institutions. Duties include development/modification of techniques for preparation of biological tissues for quantitative elemental microanalysis, operation of the LLNL microprobes to analyze prepared biological samples, X#030#ray data reduction and analysis, planning/designing and executing independent and collaborative research projects and publication of results in peer#030#reviewed literature. Requires a recent Ph.D. in chemistry, biochemistry, toxicology, pharmacology or related field. Experience in trace element analysis and analytical microscopy is desired. LLNL offers a challenging work environment and a competitive salary/benefits package. Qualified individuals are invited to send their resume to: Lawrence Livermore National Laboratory, Attn.: Mary Anne Holman, L#030#725, P.O. Box 5510, R#030#4249, Dept. AJSC527MH, Livermore, CA 94551#030#5510. LLNL is an Equal Opportunity Employer. Lawrence Livermore National Laboratory
Message-Id: {3.0.1.32.19970612093328.006c7374-at-pop3.unsw.edu.au} X-Sender: s7001031-at-pop3.unsw.edu.au X-Mailer: Windows Eudora Pro Version 3.0.1 (32)
At 11:45 AM 6/11/97 +0200, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Osmium which has been reduced can be reoxidised with hydrogen peroxide. Add it drop by drop until the light straw colour of OsO4 is restored. I cant see why it shouldn't be as good as new afterwards. Hydrogen peroxide is also good for wshing out a bottle before making up OsO4 solution in it.
Mel Mel Dickson President, Australian Society for Electron Microscopy Director, Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
We are looking to purchase a used Carbon evaporator for SEM, EPMA work. The Denton DV-502 or 502A is a desired unit. The Denton Bench Top Turbo unit would also be highly considered. Any information would be greatly appreciated.
Thank You Harry A. Ekstrom AlliedSignal Engines M/S 46-00/302-101 P.O.Box 52181 Phoenix, AZ 85072-2181 602-231-2744
It is very easy to distinguish collagen fibers from elastin in routinely fixed and stained biological samples. Any atlas of ultrastructure (e.g. Johannes A. G. Rhodin, etc.) will have typical examples of each. Basically collagen in longitudinal section displays a characteristic 640 Angstrom banding pattern while elastic fibers have an amorphous central substance surrounded by a peripheral mantle of microfibrils. In cross-section, the two components of elastin will be visible while collagen will appear to be uniform (protein). At least that's the way I remember them from my connective tissue research days.
I was wondering if anyone knows where I can get a copy of the Transmission Electron Microscopy Monograph series by Edington. I believe they are out of print but I was hoping that there may be a place that still has a supply of them. Thanks.
Your problems with wrinkels- can be caused through many ways Are you sectioning with glass or diamond knife? It is possible to eliminate wrinkels by vapours of organic solvent (xylene, chloroform, trichloroethylene, etc) to do this for example, a sheet of filter paper or a wooden applicator stick saturated with the solvent is held as close as possible above the sections on the water surface, but without touching the sections. this should expand the sections, and get rid of wrinkels. I found chloroform to be more successful.
If you cannot expand with all solvents available - than use heat. to do this suitable wires are needed, can be bought commercially. You can use a wire loop, used for picking cryo sections 2mm diameter. Heat wire with a bunsen burner (until red hot) and than wave over sections. Plastic sections have the advantage that they can be spread by heat.
If the above does not work, than check your knife height, knife angle -check to see if specimen block , knife etc is tightened. - Specimen face propely trimmed (small face) -if you are sectioning with glass knife, try tungsten coated knife or diamond knife. ( clean knife, dirty knife will give you this problem) -If wrinkels still persists, pick up sections onto grids and view. It sometimes expands under the electron beam. - Check your epon resins it might be soft. - polmerization for epon resin in our lab. we cure for 48 hours at 70 degrees centigrate. I have vast experience in sectioning all types of resins, if your problem still exists, you are most welcome to contact me and we can discuss your problems.
I do hope some of my contributions will help to sort out your problems.
Best of luck
Vijay H Bandu Centre for Electron Microscopy University of Natal Private Bag X01 Scottsville 3209 South Africa
} Question #2: A client of mine wants to see if she can see the } difference between elastin and collagin at the TEM level {she is } looking for the relative amounts in various samples}. Is this } possible? What are the references/protocols? Am I nuts to try this? } } Thanks, } John Grazul
I've seen one response to this question already, which points out quite rightly that elastin and collagen are both quite distinctive in appearance in the TEM, and so they can't be confused with one another.
However I suspect John's problem is that neither collagen nor elastin stains well with the usual U acetate/Pb citrate combination. Here are our methods (cheap!):
For elastin - Make up 0.2% orcein in acid alcohol (0.1g orcein, 50ml 70% ethanol, 0.3ml conc. HCl) and filter. Keeps well. Stain grids on drops of this stain (e.g. on a wax sheet, covered) for 30 mins at room temp., then rinse with 50% ethanol and stain with Ua and Pbc as usual.
We use this to demonstrate the elastin in artery walls, and both newly-forming elastin and "old" elastin stains well. I don't have a reference for this method, I'm afraid ("Ancient lecipe, rost in time...").
For collagen - Make up 0.01% tannin (tannic acid) in water, preferably fresh, and stain grids on drops of this for 3 mins at 60 deg.C. Rinse with water and then stain with Ua and Pbc as usual.
Collagen shows up well, but so do some other extracellular components including elastin sometimes. The reference is: Dingemans et al. (1990) Ultrastructural Pathology ,vol.14, 519-527.
N.B. - We always post-stain with Ua and Pbc because we want to see other features in the sections, and it is possible orcein and tannin won't work on their own. So if you want to stain only collagen and elastin, and nothing else, you might have to resort to immunogold methods.
Regards
Stephen Edgar
Electron Microscope Unit, Pathology Department School of Medicine University of Auckland Private Bag 92019 Auckland New Zealand
If you are using coverslips of 13mm size you can process them in 24 well tissue culture plates.. Else the 12 well plates for larger ones.
Neelima Shah.
At 11:29 AM 6/11/97 +0200, Reinhard Windoffer wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear All: Steve Barlov wrote about a used Zeiss 9 from San Diego. Although I do not know this particular EM, I am quite familiar with this type. Here are some details: It is some 25 years old, has a max mag of about 60k and 60kV accelerating voltage. The camera system is automatic. It requires little space and is contained in one unit. The electronic still uses tubes. It lend itself excellent for lower resolution work or student training. If you need more info or any other help please contact me. Peter Jordan
Dear Coordinator, I write to confirm that I have received and read your mail concerning the Microscopy forumand that my particulars are correct. i will therefore be gratefull if you finally include me on your list.
I have had several years' experience of working with hardwood tissues in studying cambium and vascular differentiation (in Aesculus hippocastanum) for routine TEM and immunolocalization of cytoskeleton and cell wall and plasmalemma epitopes. If you have access to a library, can I suggest you look at Protoplasma vol 197, pages 64-75 (1997)(Chaffey et al. TEM, JIM-staining and cytoskeleton localization) and Int. J Plant Sci. vol. 158, pages 97-109 (1997) (Chaffey et al. TEM, Thiery staining, ZIO-staining and JIM-staining) for protocols and pictures? Also look at the two recent papers by Farrar and Evert in Trees vol. 11, pages 191-202 and 203-215 (1997) (so I don't just quote my own work!)
In short, for TEM I use 2.5% glut, 3.7% form in 25 mM PIPES buffer, pH 6.9, Os post-fix, alcohol dehydration, prop oxide transfer and Spurr resin. For JIM-staining, I use same primary fix, alcohol dehydration and LR white embedding either UV-cured at -20 C, or at c. 45 C. I have not yet got a TEM localization method for tubulin or F-actin in this system (although I can supply details of indirect immunolocalization of these components at the light level).
I hope some of the above is useful. If you want to know/ask more, please contact me direct (please supply fax number(s) if protocols required: I just can't get the hang of sending 'attachments'!).
Good luck!
Nigel Chaffey
----------------------------------------------------- Dr Nigel Chaffey, Dept Forest Genetics & Plant Physiology, Swedish University of Agricultural Sciences, SE-901 83 Umea, Sweden Phone: +46-90-166305 Fax: +46-90-165901 eMail: nigel.chaffey-at-genfys.slu.se
Excuse me about my short question. I believe that I have problems with the osmium because I haven't got that problem when the tissue is not osmicated. I buffered the osmium solution with phosphate and cacodylate buffer and the osmium precipitates appear in the sample (using different animals tissues, I haven't got the problem in plant cells). The protocol is as follow: .fixation: 0-3% paraformaldehyde and 1-2.5% glutaraldehyde in phosphate or cacodylate buffer 0.1M pH 7.4. Time: 30'- 12 h. Temp: Room temp. or 4C. .Rinse with the same buffer solution.Time: 4-5 x 10'. Temp: Room temp. .Postfixation: 1-2% osmium tetroxide buffered with the same buffer solution. Time: 1 h. 4C .Rinse with water (before the osmium precipitates problem we rinse with buffer). Time: 4 x 10'. Temp: room temp. .Dehydration : acetone .Embedding: Spurr, Araldite..
The last test is buffer the osmium solution with veronal acetate but I fhaven't got the results yet.
Thanks very much for your help
Nuria Cortadellas Department of Electron Microscopy University of Barcelona
I just read the FTIR postings and know nothing about this method. I note that they refer to particulates but can it be used to identify hydrocarbons? With pollution in mind. I remember years ago that I kept a leaflet about an instrument which could do this using very small samples (on microscope slides, if I remember correctly). Now, I can't find the leaflet!
I may have inadvertently sent two messages to the list that were intended for indivviduals. To compound the error they both have hefty (280k) attachments. Please accept my apologies.
On Wed, 11 Jun 1997 Robert414-at-aol.com wrote: About 5 years ago it was common for our Taiwanese students to have a locally printed book containing all five volumes of Edington. I understand that at that time in Taiwan the laws of copyright were different from in Britain.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I was wondering if anyone knows where I can get a copy of the Transmission } Electron Microscopy Monograph series by Edington. I believe they are out of } print but I was hoping that there may be a place that still has a supply of } them. Thanks. } Alasdair Preston Dept of Mat Sci and Met Cambridge University Pembroke Street Cambridge
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Reply to: RE} osmium precipitates
Dear Nuria, Sounds like your phosphate buffer interacting with the osmium may be the cause of precipitates. We generally fix for resin embedding in cacodylate buffer. If phosphate buffer is used for the fixative, we wash it away 3 x 10 minutes with cacodylate buffer then proceed with osmication. The osmium is also never diluted in phosphate buffer. We keep a stock solution of 4% made in water then dilute to 2% with cacodylate buffer when ready to use. Hope this helps. Linda Chicoine Center for Cell Imaging Dept. of Cell Biology Yale University 333 Cedar Street New Haven, CT 06520-8002 203-785-3646 tel. 203-785-7226 fax
--------------------------------------
Excuse me about my short question. I believe that I have problems with the osmium because I haven't got that problem when the tissue is not osmicated. I buffered the osmium solution with phosphate and cacodylate buffer and the osmium precipitates appear in the sample (using different animals tissues, I haven't got the problem in plant cells). The protocol is as follow: .fixation: 0-3% paraformaldehyde and 1-2.5% glutaraldehyde in phosphate or cacodylate buffer 0.1M pH 7.4. Time: 30'- 12 h. Temp: Room temp. or 4C. .Rinse with the same buffer solution.Time: 4-5 x 10'. Temp: Room temp. .Postfixation: 1-2% osmium tetroxide buffered with the same buffer solution. Time: 1 h. 4C .Rinse with water (before the osmium precipitates problem we rinse with buffer). Time: 4 x 10'. Temp: room temp. .Dehydration : acetone .Embedding: Spurr, Araldite..
The last test is buffer the osmium solution with veronal acetate but I fhaven't got the results yet.
Thanks very much for your help
Nuria Cortadellas Department of Electron Microscopy University of Barcelona
------------------ RFC822 Header Follows ------------------ Received: by QuickMail.Yale.edu with SMTP;12 Jun 1997 03:40:34 -0400 Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id CAA03478 for dist-Microscopy; Thu, 12 Jun 1997 02:14:01 -0500 Received: from giga.sct.ub.es (giga.sct.ub.es [161.116.19.30]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id CAA03475 for {Microscopy-at-Sparc5.Microscopy.com} ; Thu, 12 Jun 1997 02:13:58 -0500 Received: from localhost (nuriac-at-localhost) by giga.sct.ub.es (8.8.0/8.8.0) with SMTP id JAA05570 for {Microscopy-at-Sparc5.Microscopy.com} ; Thu, 12 Jun 1997 09:11:47 +0200
A student asked me to query about the density of Fe3Si. I did not see it in the phase diagram available to me.
Thus does it exist and if it does do some have a measured density for it?
Thanks
## [########] ## ## ## ## ## Stephan H Coeztee Electron Microscope Unit Private Bag 3 Wits 2050 South Africa
I have always tried to avoid cacodylate buffer by using safer alternatives such as phosphate or Pipes but I am about to embark on some new work where all of the references have used cacodylate in the past. Obviously it is good practice to try and reduce the use of hazardous materials in the lab., especially in e.m., where we have so many. This is embodied in our safety laws in the UK especially in COSHH regulations (Control of Substances Hazardous to Health 1988) but, judging by recent worries on the list about formaldehyde and glutaraldehyde, is a principal that we all adhere to.
My question is: has anyone come up with a safer alternative to cacodylate that could be used as a direct substitute? Phosphate won't do because of its problems with Ca++ but Pipes seems safer (the safety data sheets I have are not as helpful as they could be), although it is expensive and probably won't keep as long.
A student from Botany is experiencing problems with staining cassava somaltic embryos at the globulas stage. She is embedding in Spurs Resin. Reynold's staining is used with the staining times: 30 min uranyl acetate 1 hr in lead staining
The staining is very week.
## [########] ## ## ## ## ## Stephan H Coeztee Electron Microscope Unit Private Bag 3 Wits 2050 South Africa
We have a DV-502 which we use mostly for TEM samples. In general it works just fine but you might want to keep in mind the following :
1)The carbon rod is spring loaded, and the spring that we purchased from Denton is quite stiff. This tends to cause a premature brake of the carbon tip. I would recomend changing this spring to one that is less stiff.
2) With time the bell jar and the various components within the chamber will get cabon coated and will have to be cleaned. Depending on the level of coating this can take quite some time. I would suggest that you modify the shield ( i.e. build a larger one ) that will result in less carbon build up in the surrounding areas and which will be easier to clean.
Jordi Marti.
P.S. Dending on the kind of work/samples a bench top model (evaporator and sputter unit) with a mechanical pump might be sufficient for your work and you might save some $$$. -----------------------------------------
Although Jan may know of some cache at PEO with these books (and different Philips personnel have different amounts remaining), my contacts with Philips over the past few years to obtain these volumes have indicated that the only remaining copies are the least requested volumes. The two (or three?) most used volumes have been out of stock for some time now. However, the volumes have been combined into a single volume and are still in print, or at least available, (although in a *much* poorer quality) from TechBooks. Techbooks is at 4012 Williamsburg CT Fairfax, VA 22032-1139 (703-352-0001). (I just found another Techbooks listing at: Campus Square Klamath Falls, OR 97601 (541-884-3882) but don't know if they would also have Edington's book.)
I have also seen a number of versions printed in Asia, but even aside from the copyright concerns, the quality of the ones I've seen has been no better (and may have been worse) than the TechBooks version.
Good luck in your search.
Dick Fonda
In message {39f71d60-at-pei.philips.com} jan ringnalda writes: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } The Edington books are still available through Philips Electron Optics } although supplies now are getting limited. } } Cheers, Jan
_____________________________________________________________ Richard W. Fonda Naval Research Laboratory (202) 767-2622 Code 6324 (202) 767-2623 fax Washington DC 20375 _____________________________________________________________
At 04:13 PM 6/11/97 EDT, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America It would all depend on the heat load being produced by the instruments that you are trying to cool. Check with your manufacturers. We once considered using a single Haskris with a "Y" in the supply line to serve two instruments with low heat load at the same time, while we were waitng for repair parts for the other Haskris. Hitachi service man though it should work. Ultimately we were able to get along without resorting to that, so I can't say for sure if it would work.
} Question #2: A client of mine wants to see if she can see the } difference between elastin and collagin at the TEM level {she is } looking for the relative amounts in various samples}. Is this } possible? What are the references/protocols? Am I nuts to try this?
Could antibodies be used to differentiate the two in some way } } Thanks, } John Grazul } Rutgers University } Electron Imaging Facility } } ******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
thanks to everyone who replied to my two very different questions. The collagen/elastin experiment will start next week if the researcher can get the samples, and the chiller will be retrofitted ASAP.
John Grazul Rutgers University Electron Imaging Facility
The purity of your glutaraldehyde may be the problem. At one time we re-distilled ours to avoid the "pepper" problem. We still sometimes have a the problem you describe especially in muscle. It appears from time to time for no explicable reason.
} } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } At 09:11 AM 6/12/97 +0200, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
Here in Reading, we are still using a Philips 301 TEM with a 35 mm camera. Until about a year ago, we were using Agfa Scientia 35 mm unperforated roll, when this was discontinued. Many people have offered us Eastman 5302, but this has two disadvantages (1) it is only about 25% as sensitive to electrons (2) it suffers stress marks when packed too tightly into the cassette.
We have already contacted microscopy groups on an individual basis, from Finland in the North to New Zealand in the South, and it appears this situation is global. If you have any ideas, please contact us IF:
(a) you know of another source of film; (b) you would like to see 35 mm Agfa Scientia, and would like to form a group to approach the company.
Please don't reply simply to say "we use plates", on the principle:
"I only want a little bit of butter for my bread" "But many people nowadays like marmalade instead"
(from A.A.Milne)
Thank you
+------------------------------------------------------------------------+ | 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 do not like Spurr Resin because it gives inconsistent results. Sometimes it stains poorly. Try Epon 812 or its substitutes or LR White. Lead staining for 1 hr seems too long and it is likely to precipitate. I usually stain for 5 min. on Epon sections.
A student from Botany is experiencing problems with staining cassava somaltic embryos at the globulas stage. She is embedding in Spurs Resin. Reynold's staining is used with the staining times: 30 min uranyl acetate 1 hr in lead staining
The staining is very week.
Stephan H Coeztee Electron Microscope Unit Private Bag 3 Wits 2050 South Africa
I saw your posting on the listserver and just wanted to re-send my message dated 6/5/97 in case that you did not receive it.
Dear Richard:
Since you are getting faint lattice images from the YBCO sample that we prepared, it is quite possible that the carbon film is the source of scattering. The "conventional" method of FIB preparation requires that the specimen is mounted on a grid, but a carbon support is not needed, that is, the electron beam will be passing through the electron transparent area only. We can try to prepare the YBCO film using conventional methods of FIB specimen preparation. We might be able to try it with the remaining sample, but this technique might require a larger piece of sample. Can you send us an additional piece?
Can you also please forward the lattice parameters of YBCO to me so that we can test and view the specimen for lattice images prior to shipping it to you?
If this specimen works out, we will charge you the previously quoted rates for TEM FIB specimen preparation:
TEM Sample Preparation:
FIB sample preparation $250/hr other sample preparation $150/hr
TEM work $200/hr
I anticipate that the preliminary specimen preparation might require up to 5 hours, FIB usage will probably be about 3 hours with no more than 5 hours of use. The TEM will be used for ~ 1 hour to check for the usefulness of the specimen. Based on these figures, the specimen can be prepared for ~ $1,700 - $2,200. If this is something that you would like to pursue please let me know at your earliest convenience.
Thank you,
Lucille Giannuzzi
************************************************************************* Lucille A. Giannuzzi, Ph.D. Assistant Professor
Dept. of Mechanical, Materials, and Aerospace Eng.
University of Central Florida phone (407) 823-5770 PO Box 162450 fax (407) 823-0208 4000 Central Florida Blvd. email lag-at-pegasus.cc.ucf.edu Orlando, FL 32816-2450 USA *************************************************************************
Dear Microscopists, Our laboratory urgently needs formula for preparation Lowicryl K4M resin for low temperature embedding after freeze-drying. We have all set of chemicals in hand, but without any precise instruction how to mix them. Thanks in advance for all help Best regards Jolanta Przybylowicz
Independent from the fixative I would consider using Lowicryl HM20 instead of LR White, its viscosity is much lower and immuno labelling works nicely. For example aleurone cells of barley grains (which have huge cell walls, which even represent a problem with Spurrs) are easy to infiltrate with this stuff at -35 C, and organells are preserved quite well.
Good luck, Stefan
Dr. Stefan Hillmer Albrecht-von-Haller Institut fuer Pflanzenwissenschaften Universitaet Goettingen Untere Karspuele 2 37073 Goettingen Deutschland
Tel (+49) 551-392013 Fax (+49) 551-397833 e-mail shillme-at-gwdg.de
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I will be involved with operating an ESEM in the near future and need } information about any courses available concerning this technology. Any } information will be greatly appreciated. } } Well Gary, I don't know where you are at Geographically, - But if anyone is interested - The Institute of Meteoritics at the University of New Mexico holds an EM course once a year that includes teaching on the EMP, SEM, ESEM, and LVSEM, as well as sample prep, limitations, new technology and an overview of Secondary Ion Mass Spec. (SIMS). We also teach courses in TEM, ICPMS, and a number of other types of equipment (XRD, OM...etc...). Of course the problem is that you have to get to New Mexico to take the course.
For More Info, reply to yoyodine-at-unm.edu
I would suggest to anyone looking for courses in Microscopy of any kind to contact the nearest University...classes of this type or offered in depts. such as Materials Science, Earth and Planetary Science, Geology, Biology, Chemistry, and Engineering...SEMS are rather common and ESEM are basically the same machine with the addition of a gated vacuum system...any hands on SEM course should do the trick.
Hope this helped Christopher Adcock
PS: I replied to all on the list because I thought others might be interested, but I have noticed that I get alot of clutter in my mbox and others most likly do to. So if you want more info, it would be best to reply to me alone and keep others boxes clean.
In some cases, cacodylate is the preferred buffer because it helps the fixative to penetrate the tissue faster and further. Certainly, it is hazardous, but if one uses a fume hood and wears gloves one can minimise the danger, and of course you should be doing this anyway, since the fixative itself is so much more dangerous than the cacodylate.
Lesley Weston
On Thu, 12 Jun 1997, HASWELL Malcolm wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Dear all } } I have always tried to avoid cacodylate buffer by using safer alternatives } such as phosphate or Pipes but I am about to embark on some new work where } all of the references have used cacodylate in the past. Obviously it is good } practice to try and reduce the use of hazardous materials in the lab., } especially in e.m., where we have so many. This is embodied in our safety } laws in the UK especially in COSHH regulations (Control of Substances } Hazardous to Health 1988) but, judging by recent worries on the list about } formaldehyde and glutaraldehyde, is a principal that we all adhere to. } } My question is: } has anyone come up with a safer alternative to cacodylate that could be used } as a direct substitute? Phosphate won't do because of its problems with Ca++ } but Pipes seems safer (the safety data sheets I have are not as helpful as } they could be), although it is expensive and probably won't keep as long. } } thanks in advance, } } Malcolm Haswell } University of Sunderland } UK } } }
} Excuse me about my short question. I believe that I have problems with } the osmium because I haven't got that problem when the tissue is not } osmicated. I buffered the osmium solution with phosphate and cacodylate } buffer and the osmium precipitates appear in the sample (using different } animals tissues, I haven't got the problem in plant cells). The protocol } is as follow: } .fixation: 0-3% paraformaldehyde and 1-2.5% glutaraldehyde in phosphate or } cacodylate buffer 0.1M pH 7.4. Time: 30'- 12 h. Temp: Room temp. or 4C. } .Rinse with the same buffer solution.Time: 4-5 x 10'. Temp: Room temp. } .Postfixation: 1-2% osmium tetroxide buffered with the same buffer } solution. Time: 1 h. 4C } .Rinse with water (before the osmium precipitates problem we rinse with } buffer). Time: 4 x 10'. Temp: room temp. } .Dehydration : acetone } .Embedding: Spurr, Araldite.. } } The last test is buffer the osmium solution with veronal acetate but I } fhaven't got the results yet.
If freshly depolymerized paraformaldehyde is not used, the short formaldehyde 'mers are left behind in your tissues and react with the osmium to form granular precipitates (very small, peppery looking).
Phosphate buffers are notorious precipitators (especially with uranyl salts) but cacodylate should be precipitate free.
In some (perhaps a lot of) instances it is not necessary to buffer the osmium - simply use an aqueous solution.
Is your distilled water good? Water containing volatile amines or other organics may react with the osmium/aldehydes to cause precipitates.
Please describe the precipitates (large, anglular, blobs, everywhere, localized, etc).
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
I am trying to locate a laboratory or University that has Scanning White Light Interferometry available for contract research. The two instruments I am familar with and have provided us with useful data are manufactrued by Zygo and WYKO.
Thanks, John Catino Union Camp Corp. Princeton, NJ
Absolutely - Actually, I think I can speak for the other respondents in saying that most of the particles and fibers that we are referring to are organic in nature. Organic compounds (i.e. various forms of hydrocarbons) are the primary analytes examined by FTIR. Generally speaking, depending upon the sample condition and matrix, one can obtain very good qualitative and even quantitative FTIR assays. CH absorptions abound in the following regions of the spectrum (in wavenumber):
2800-3000 cm-1 1300-1490 cm-1
If you were to collect airborne hydrocarbon particulates onto filters, you could qualitatively analyze these by micro-FTIR - you may not get a positive identification of a single compound (because you are likely to have a mixture), but you might be able to identify chemical families.
Regards,
Bob ************************* Bob Citron Chiron Vision Claremont, CA USA (909)399-1311 Bob_Citron-at-cc.chiron.com **************************
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello from Plymouth, UK
I just read the FTIR postings and know nothing about this method. I note that they refer to particulates but can it be used to identify hydrocarbons? With pollution in mind. I remember years ago that I kept a leaflet about an instrument which could do this using very small samples (on microscope slides, if I remember correctly). Now, I can't find the leaflet!
Just want to thank all of you for your responses to my question on xylene replacements. I now have a number of leads which I will follow up on shortly.
Paula.
Paula Allan-Wojtas Food Microstructure Specialist Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
To get good electron channeling patterns you need a surface that is completely free of any form of mechanical distortion such as can be introduced by most mechanical cutting, grinding and polishing operations. If you must use mechanical techniques to prepare your specimen's surface, then you must etch it rather heavily after each stage(after cutting, after grinding, and after each polishing step) to remove as much cold-worked metal as possible, and at the end you must use a much heavier etch than you normally would for just microscopic analysis - you must see clearly defined, distortion-free grains. This will require some experimentation to find an etchant that will remove lots of metal without pitting. If possible, it is preferable to use electrolytic polishing, because this does not introduce mechanical deformation. To get started, might I suggest getting a piece of single crystal silicon of the type used in the semiconductor industry, which should give good channeling patterns as-received, and play around with that until you get the hang of the technique.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
I have found dimethylsulfoxide (DMSO) to be a truly remarkable material to use for treating minor burns of the ordinary variety. I have found all the claims made on p. 107, and elsewhere, in the book 'DMSO, The Pain Killer' (by Barry Tarshis, Berkley Books, 1981, ISBN: 0-425-07488-9) to be unbelievably true. The minute you experience a common-type, 'small' burn, wash it for a few minutes with cold water, then apply a 70% DMSO-30% water solution libally immediately, and then two or three more times at about one-hour intervals, and then again twice daily for a couple more days. I find this procedure will usually stop the pain almost immediately, and more surprisingly, will usually keep the burn from blistering. I have found it also works if you begin to develop a blister from having a shoe rub your heel, or from raking the lawn without gloves, etc.- prompt and persistent application of DMSO will usually relieve the pain and keep a blister from developing. I haven't tried it for LN2 burns, but would sure give it a try if I happened to experience one. I keep a bottle in my office, and another at home, for just such occurrences.
Dimethlysulfoxide is not a material that is approved for medical use, and so if you make use of it you do so under your own responsibility; however, it has long been used by athletes to relieve the pain from bruises and sore muscles, and it is reported (see above source) to also be effective in relieving the pain associated with some cases of bursitis, arthritis, interstitial cystitis, scleroderma, headaches, gout,hemorroids, herpes infections, shingles, etc., etc.
DMSO is not medically approved because it is a cheap by-product of the pulp and paper industry, and therefore not patentable as a drug. Thus, no drug company is willing to go to the expense of running the necessary test to get it approved. In addition, most people develop a garlic-like taste from using it, and so it is almost impossible to run the double-blind tests needed to gain such approval. It is, however, widely available in health food stores.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
Many thanks for your kind input regarding my PBS argument. I have decided to use a slightly hypertonic (4mM) phosphate buffering system in 9% saline solution. The slight hypertonicity (and slightly increased buffering capacity) may help preserve tissue during the DAB reaction, because this tissue still has semi-permeable membranes due to the absence of osmium fixation. Thanks again. This Listserver is better than chocolate! Bye, Hildy
john catino wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } I am trying to locate a laboratory or University that has Scanning White } Light Interferometry available for contract research. The two } instruments I am familar with and have provided us with useful data are } manufactrued by Zygo and WYKO. } } Thanks, } John Catino } Union Camp Corp. } Princeton, NJ } } -- } II*Dear John,
Both Zygo and Wyko have facilities here in the Northeast which are available on a contract basis. Call Zygo at (460)247-8372 (Ask for Barbara - not me) and Wyko at (602)741-1297.
} } P.S. There was a thread awhile ago on what gloves to use with what } } chemicals--was there a definitive list that came out of that discussion? I } } know glut goes through latex like the glove isn't there. P } } } } } Sic Hoc Legere Scis Nimium Eruditionis Habes { } } Philip Oshel
Phil and anyone else who is interested, I spent some time researching the issue of what gloves our Unit should be recommending for all our users. As a result I drew up a table of which gloves to wear when handling various common chemicals in our Unit. I can send anyone interested a copy of this, however I take no responsibility for the information contained in it in any way whatsoever (except to people using our EM Unit), I'm still updating it as I learn more.
The whole subject is more complex than I first thought. There is quite a bit of apparently contradictory information on glove suitabilty, largely resulting from the large number of different glove polymers, glove styles, chemical combinations and work practices around. Because we are a multiuser lab with about 90 users on our books at present we really were only interested in disposable gloves. Thick, heavy gloves that hindered fine manipulation were also out. I came to the conclusion that, with a few important exceptions, latex gloves are actually fairly good for most of the chemicals we used based on the studies I read.
Although latex has a poor reputation for use with glutaraldehyde there is at least one paper (Jordan et al, Glutaraldehyde permeation: Choosing the right glove, Union Carbide 1996) which guardedly suggests that it is probably OK for use with the low concentrations of glutaraldehyde usually used for fixing tissue, as long as contaminated gloves are removed as soon as possible. That really is one of the crucial aspects of wearing gloves for chemical protection - all gloves are permeable eventually to many chemicals and one of the best ways to minimise exposure risk is to remove gloves as soon as possible after contamination and don new ones. In the above paper the authors state that the breakthrough time for a single layer of latex examination gloves exposed to 2% glutaraldehyde was between 30 and 45 minutes (at 25degC however). This is possibly good enough for people dealing with specimens in low concentrations of glutaraldehyde, provided they don't continue to wear the gloves when contaminated and provided the individual is not sensitised to glutaraldehyde. Therefore it is probably not true to say that 'glut goes through latex like the glove isn't there', it does provide short-term protection. Whether it provides enough is another matter, it may be possible that latex permits enough glutaraldehyde through to eventually cause sensitisation to glutaraldehyde. If you want to be safer, nitrile and butyl synthetic rubber provide a much better barrier than latex.
I think that if you want to minimise the risk you should use nitrile instead of or as well as (over) latex for glutaraldehyde. I personally get slightly itchy hands from using our disposable nitrile gloves so I tend to wear them over latex gloves. Nitrile is also supposed to be better as a barrier for acrylic resins too. You can't substitute disposable nitrile gloves for latex totally however because nitrile doesn't cope with exposure to some resins and solvents. Nitrile is particularly ineffective as a barrier against propylene oxide (less effective than latex anyway - however latex isn't too good against propylene oxide either).
You can apparently get extra protection by using one of the spray-on barrier creams too.
Regards,
Richard
Richard Easingwood South Campus Electron Microscope Unit School of Medical Sciences University of Otago PO Box 913 Dunedin NEW ZEALAND
Subject: Time:4:25 PM OFFICE MEMO Need EM200 sample holder Date:6/12/97
Hello All,
We're looking for an inexpensive TEM sample holder for the Philips EM200 or EM 400 series microscopes. Used, simple, single tilt, is fine, if not bent. Key parameter is inexpensive.
DGCollins-at-lbl.gov (510) 486-7859, or DCollins 2841 Kinney Dr, Walnut Creek, CA 94595 (510)939-2006
*Received: from SpoolDir by IM-PW (Mercury 1.13); Thu, 12 Jun 97 23:33:05 -1800 *Return-path: {Microscopy-request-at-sparc5.microscopy.com} *Received: from Sparc5.Microscopy.Com by mailer.inmat.pw.edu.pl (Mercury 1.13); * Thu, 12 Jun 97 23:32:54 -1800 *Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id PAA06007 for dist-Microscopy; Thu, 12 Jun 1997 15:00:13 -0500 *Received: from srvr7.engin.umich.edu (srvr7.engin.umich.edu [141.212.2.69]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id PAA06004 for {microscopy-at-sparc5.microscopy.com} ; Thu, 12 Jun 1997 1
*:00:12 -0500 *Received: from [141.212.131.74] (dow2146amac74.engin.umich.edu [141.212.131.74]) * by srvr7.engin.umich.edu (8.8.4/8.8.4) with ESMTP * id QAA25518 for {microscopy-at-msa.microscopy.com} ; Thu, 12 Jun 1997 16:00:00 -0400 (EDT) *X-Sender: bigelow-at-srvr5.engin.umich.edu *Message-Id: {v03007802afc5bce8e0dc-at-[141.212.131.74]} *Mime-Version: 1.0 *Content-Type: text/plain; charset="us-ascii" *Date: Thu, 12 Jun 1997 10:58:02 -0400 *To: Microscopy Listserver {microscopy-at-Sparc5.Microscopy.Com} *From: Wil Bigelow {bigelow-at-engin.umich.edu} *Subject: RE:Electron Channeling
Professor is absolutly right. There should be also kept in mind that the investigated material should be free of cold work in order to obtained sharp channeling patern. In the past, there was done a lot of work by Prof. W.W.Gerberich and members of his research group on influance of strain on channeling pattern.
Witold Zielinski Warsaw University of Technology Narbutta 85 02-524 Warszawa, Polska
I am interested in gloves resistant particularly to acetone, which most of our dehydration is done in.
Having seen the initial posting, I tried a crude test today on some new, thin nitrile gloves: 15 minutes dangling the finger ends (empty!) in conc. HNO3, 25% glut and pure acetone (in separate beakers). Then the gloves were washed carefully on their exterior and dabbed dry with a towel. The HNO3 trial showed a serious failure problem! The other gloves were then blown up by mouth and tested with the Mark I nose. The acetone penetration was serious, but the glut was not detectable (and this Mark I nose is normally quite sensitive). That's it.
} From: "Robert H. Olley" {R.H.Olley-at-reading.ac.uk} } To: Microscopy Newsgroup {Microscopy-at-Sparc5.Microscopy.Com} } Cc: # {R.H.Olley-at-reading.ac.uk} } Subject: TEM: 35 mm film - where to find? } } Here in Reading, we are still using a Philips 301 TEM with a 35 mm } camera. } Until about a year ago, we were using Agfa Scientia 35 mm unperforated roll, } when this was discontinued. Many people have offered us Eastman 5302, but } this has two disadvantages (1) it is only about 25% as sensitive to } electrons (2) it suffers stress marks when packed too tightly into the } cassette. } } We have already contacted microscopy groups on an individual basis, from } Finland in the North to New Zealand in the South, and it appears this } situation is global. If you have any ideas, please contact us IF: } } (a) you know of another source of film; } (b) you would like to see 35 mm Agfa Scientia, and would like to form a } group to approach the company. } Thank you } } +------------------------------------------------------------------------+ } | 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 | } +------------------------------------------------------------------------+ } Dear Mr. Olley,
Many of our M. users have already changed to Kodak 35 mm Technical Pan Film and are very positive about it. However with respect to Agfa Scientia and Kodak 5302 it has two disadvantages: it is perforated and you need a special developer.
Regards,
Bert Boxstart Philips Electron Optics B.V. Comm. Support dep.
Hi I am urgently looking for the E-mail address of either Dr DJ Cork or J P Krueger from the Illinois Institute of Technology in Chicago. If you can be of assistance please reply to my E-mail address.
I apologize for this non-related microscopy request
Thank you Alan N Hall Unit for Electron Microscopy Faculty of Biological & Agricultural Sciences University of Pretoria Pretoria 0002 Republic of South Africa Tel: +27-12-420 3297 Fax: +27-12-420 3266
Sorry this is so late in the train of information here, but I have been told by two or three technicians who use DAB regularly to use Tris buffer instead of phosphate buffer with the DAB solution. One even had me doing a Tris buffer rinse after a fix with PBS to make sure there was no phosphate buffer remaining at the DAB-labeling step. I forgot what the problem was, but maybe someone else out there can add some info. The point is, you may have solved the tonicity problem, but you could still have some problem with the DAB substrate reaction if you are using PBS with the DAB. P.S. I got the recipe for Tris buffer from Handbook of Immunoperoxidase Staining Methods by Janice A. Bourne, published by DAKO Corp. in 1983. Tris buffer (0.05M) is 6.1g trishydroxymethyl aminomethane (tris base), 50 ml dH2O, mix and add 37 ml 1N HCl, dilute to 1 liter with dH2O. The pH should be 7.6+/- 0.2 at 25oC (I often use a pH or 7.2, just adjust the volume of HCl). They also have a recipe for Tris buffered saline using 100ml of this Tris with 900ml of 0.85% saline or PBS. They say it can reduce background staining.
Another way to help preserv tissue at the DAB step is to include a non-peroxide DAB step prior to the actual reacting step. I find that this enhances the reaction and often cuts down the time of exposure to the peroxide. Hope this is helpful.
Karen
On Thu, 12 Jun 1997, HILDEGARD CROWLEY wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Dear Folks, } } Many thanks for your kind input regarding my PBS argument. I have } decided to use a slightly hypertonic (4mM) phosphate buffering system in } 9% saline solution. The slight hypertonicity (and slightly increased } buffering capacity) may help preserve tissue } during the DAB reaction, because this tissue still has semi-permeable } membranes due to the absence of osmium fixation. } Thanks again. This Listserver is better than chocolate! } Bye, } Hildy }
I am working with the human cornea and am exploring options for tissue preparation techniques and staining procedures for LM. Sections of 0.5-1.0 microns will be used to distinguish between the epithelium and underlying Bowman's layer. I need to find the optimal embedding medium as well as staining procedure.
I'd like to know how could I get histological specimens from titanium implants inserted into rabbit bone. The problem is cutting bone with metal. It seems that the suitable equipment is named Exact. How does it work? What about its price? Where can I get it?
Yours sincerly,
Marcelo Henrique Prado PEMM - COPPE/UFRJ Po.Box.:68505 Cidade Universitaria - Ilha do Fundao Rio de Janeiro-R.J. CEP.: 21941-900
Richard Lander wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Phil Oshel wrote; } } } } P.S. There was a thread awhile ago on what gloves to use with what } } } chemicals--was there a definitive list that came out of that discussion? I } } } know glut goes through latex like the glove isn't there. P } } } } } } } Sic Hoc Legere Scis Nimium Eruditionis Habes { } } } Philip Oshel } } Phil and anyone else who is interested, } I spent some time researching the issue of what gloves our Unit should be } recommending for all our users. As a result I drew up a table of which } gloves to wear when handling various common chemicals in our Unit. I can } send anyone interested a copy of this, however I take no responsibility for } the information contained in it in any way whatsoever (except to people } using our EM Unit), I'm still updating it as I learn more. } } The whole subject is more complex than I first thought. There is quite a } bit of apparently contradictory information on glove suitabilty, largely } resulting from the large number of different glove polymers, glove styles, } chemical combinations and work practices around. } Because we are a multiuser lab with about 90 users on our books at present } we really were only interested in disposable gloves. Thick, heavy gloves } that hindered fine manipulation were also out. I came to the conclusion } that, with a few important exceptions, latex gloves are actually fairly } good for most of the chemicals we used based on the studies I read. } } Although latex has a poor reputation for use with glutaraldehyde there is } at least one paper (Jordan et al, Glutaraldehyde permeation: Choosing the } right glove, Union Carbide 1996) which guardedly suggests that it is } probably OK for use with the low concentrations of glutaraldehyde usually } used for fixing tissue, as long as contaminated gloves are removed as soon } as possible. That really is one of the crucial aspects of wearing gloves } for chemical protection - all gloves are permeable eventually to many } chemicals and one of the best ways to minimise exposure risk is to remove } gloves as soon as possible after contamination and don new ones. In the } above paper the authors state that the breakthrough time for a single layer } of latex examination gloves exposed to 2% glutaraldehyde was between 30 and } 45 minutes (at 25degC however). This is possibly good enough for people } dealing with specimens in low concentrations of glutaraldehyde, provided } they don't continue to wear the gloves when contaminated and provided the } individual is not sensitised to glutaraldehyde. } Therefore it is probably not true to say that 'glut goes through latex } like the glove isn't there', it does provide short-term protection. Whether } it provides enough is another matter, it may be possible that latex permits } enough glutaraldehyde through to eventually cause sensitisation to } glutaraldehyde. If you want to be safer, nitrile and butyl synthetic } rubber provide a much better barrier than latex. } } I think that if you want to minimise the risk you should use nitrile } instead of or as well as (over) latex for glutaraldehyde. I personally get } slightly itchy hands from using our disposable nitrile gloves so I tend to } wear them over latex gloves. Nitrile is also supposed to be better as a } barrier for acrylic resins too. You can't substitute disposable nitrile } gloves for latex totally however because nitrile doesn't cope with exposure } to some resins and solvents. Nitrile is particularly ineffective as a } barrier against propylene oxide (less effective than latex anyway - however } latex isn't too good against propylene oxide either). } } You can apparently get extra protection by using one of the spray-on } barrier creams too. } } Regards, } } Richard } } Richard Easingwood } South Campus Electron Microscope Unit } School of Medical Sciences } University of Otago } PO Box 913 } Dunedin } NEW ZEALAND } } Telephone: 64-03-479 7301 } Facsimile: 64-03-479 7254 } } e-mail: richard.easingwood-at-stonebow.otago.ac.nz
I think it should be clearly stated that neither latex nor nitrile gloves provide any effective barrier to epoxy and acrylic resins. I became sensitized, and had a very severe reaction to acrylic resins through the misconception that nitrile gloves were the gloves of choice for handling resins. The only glove I would recommend to anyone working with resins is the 4H glove from Safety 4 A/S, 9765 Widmer, Lenexa KS 66215 (913) 492 0860. Some people may find them a bit cumbersome at first, but we usually put a pair of nitrile gloves over the 4H gloves (not for added protection, but for a better finger feel). Don't forget to also wear a protective sleeve (also 4H), as the area between the glove and lab coat is still susceptible to the fumes of the monomer. The 4H glove also has a break-through time of } 240 minutes with a 25% glutaraldehyde solution. This is my personal opinion, after much research, and suffering, and does not represent the views of Texas Scottish Rite Hospital for Children.
Ronnie Houston Histology Coordinator Texas Scottish Rite Hospital for Children Dallas, TX 75219
On Thu, 12 Jun 1997 MESJASZ-at-NACDH4.NAC.AC.ZA wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Our laboratory urgently needs formula for preparation Lowicryl K4M resin } for low temperature embedding after freeze-drying. We have all set of } chemicals in hand, but without any precise instruction how to mix them. } Thanks in advance for all help
Hi Jolanta,
Here is the K4M recipe:
Cross linked A 2.7 g Monomer B 17.02 g Initiator C 20.1 g
The dehydration can be done with a series of ethanol at a low temperature. For the infiltration, the following schedule can be carried out at a low temperature.
1 : 1 ethanol : embedding medium 1 hr 1 : 2 ethanol : embedding medium 1 hr 100 % embedding medium, 2 X 1 hr, ea.
polymerization at -30 to -40 C for 24 hrs.
Good luck
*********************************************** * Ming H. Chen, PhD * * Medicine/Dentistry Electron Microscopy Unit * * University Of Alberta. * * Edmonton, Alberta, Canada * * * * Visit My Page At: * * http://www.ualberta.ca/~mingchen * ***********************************************
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Subject: Time:7:32 AM OFFICE MEMO Post-doc position at LLNL Date:6/11/97
Our Center for Accelerator Mass Spectrometry (CAMS) is currently seeking a Postdoctoral Research Staff Member to work as part of an interdisciplinary team of physicists, chemists, and biologists. In this research position, you will work as part of a team to develop quantitative elemental analysis of biological tissues. These techniques will be used in studies of elemental kinetics, structural biology, toxicology or pharmacology by staff at LLNL and collaborators in the University of California system and other institutions. Duties include development/modification of techniques for preparation of biological tissues for quantitative elemental microanalysis, operation of the LLNL microprobes to analyze prepared biological samples, X#030#ray data reduction and analysis, planning/designing and executing independent and collaborative research projects and publication of results in peer#030#reviewed literature. Requires a recent Ph.D. in chemistry, biochemistry, toxicology, pharmacology or related field. Experience in trace element analysis and analytical microscopy is desired. LLNL offers a challenging work environment and a competitive salary/benefits package. Qualified individuals are invited to send their resume to: Lawrence Livermore National Laboratory, Attn.: Mary Anne Holman, L#030#725, P.O. Box 5510, R#030#4249, Dept. AJSC527MH, Livermore, CA 94551#030#5510. LLNL is an Equal Opportunity Employer. Lawrence Livermore National Laboratory Or contact me Graham bench Ph 510-423-5155 Cheers Graham Bench
You have unfortunately, just missed the best course on the subject. The Lehigh Short Course on SEM and Microanalysis. Contact Sharon Coe at {slc6-at-Lehigh.edu} for details od next years course.
Patrick Echlin
On Wed, 11 Jun 1997, Gary Lovell wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I will be involved with operating an ESEM in the near future and need } information about any courses available concerning this technology. Any } information will be greatly appreciated. } }
I cannot tell you how much I appreciate and enjoy your replies to my problems. Our department is rather isolated here away from the town's huge medical research center, so instead of working with a vaccum, we often work in a vaccum! I am unable to repay you all individually, but I will take the time and effort to answer any problems on the LIST SERVER on which I feel I have expert information, and in such manner hope to be helpful to some of you who have gotten me out of a corner. (Corners) Thanks again! Hildy Crowley University of Denver Denver, CO
} DMSO is not medically approved because it is a cheap by-product of the } pulp and paper industry
In a chemical lab, DMSO is a dangerous substance because it takes any chemicals you might have on your skin and carries them into your blood. Anyone who applies DMSO to their skin should do so only after washing thoroughly and then not use any chemicals until the DMSO is really gone. Also the bottle of DMSO should not be kept near any toxic chemicals, and you should be sure that no one else has contaminated it with toxic chemicals. If I were using DMSO (I am not), I would want to use a new sealed bottle each time. What if the soap contains something toxic? What if the soap fails to remove something toxic?
} Dimethlysulfoxide is not a material that is approved for medical use, and } so if you make use of it you do so under your own responsibility; however, } it has long been used by athletes to relieve the pain from bruises and sore } muscles, and it is reported (see above source) to also be effective in } relieving the pain associated with some cases of bursitis, arthritis, } interstitial cystitis, scleroderma, headaches, gout,hemorroids, herpes } infections, shingles, etc., etc.
I have found Bigelow's information interesting but it left out a very important detail about DMSO's use by athletes, especially male athletes. DMSO has the ability to neutralize testosterone. Many male athletes thrive on testosterone as a feature needed for competition. This is one of the reasons that athletic trainers quit using DMSO.
Another problem with topical application of DMSO is that it can take with it live biological materials that may be on the surface of the skin. So it can pull with it virus for example that would not normally gain access to the interior of the body. For these and other reasons DMSO has not been approved for "general" medical use.
As I read Bigelow's last sentence above, snake oil comes to mind.
Blystone in Texas
-------------------------------- Robert V. Blystone, Ph.D. rblyston-at-trinity.edu
Department of Biology Trinity University 715 Stadium Drive San Antonio, Texas 78212 210.736-7243 FAX 210/736-7229
Gustave H. Wanner wrote: ---------------------------------------------------------------------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Co ---------------------------------------------------------------------- } Hi Everyone, } } Do any of you folks have any experience using the Polaroid MicroCam } with a Nikon Labophot-2 or equivalent microscope with Color-Free } objectives? The MicroCam replaces the eyepiece of the microscope; } it is unclear to me whether or not the MicroCam includes compensation for lateral chromatic aberation (also known as chromatic difference of magnification). The Nikon (as well as other recent microscope) objectives are corrected to provide a color free intermediate image. If the MicroCam includes compensation, it seems to me that this might negatively affect the resulting photomicrographs. } I have been unable to obtain any information from Polaroid telephone technical support on this question. } } Best regards, } } Gus Wanner } Exitech Corporation } 102 E. Broadway } Maryville, TN 37804 } (423) 983-9101 ListServer-at-MSA.Microscopy.Com Gustave H. Wanner {ghw-at-EXITECH.com}
Hello Gus, I am retired now from Polaroid, but was the microscopist on the development team for the MicroCam. I investigated just the question that you raised concerning Chromatic Difference of Magnification. I tested the MicroCam on a Nikon Optiphot with F head, using a 10X/0.25 CF objective, and a stage micrometer as the target, and found no color fringing.
I tested for fringing by focusing on a stage micrometer, or on the fine grating area of an Air Force Resolution Test Target; both provided a flat, high contrast specimen. I then looked for color fringing (orange or blue) in the image near the edge of the micrograph.
In making the MicroCam, it was our goal to make an inexpensive, ($795) camera compatible with a large number of microscopes, even those without phototubes, providing through the lens viewing, and automatic exposure control, --- and ease of use. These goals led to using an eyepiece integral to the camera.
As you mentioned, the lateral chromatic abberation, also known as CDM, for Chromatic Difference of Magnification, affects the compatibility of eyepieces with objectives. CDM is the difference in magnification between red and blue images at the primary image plane. CDM occurs in the objective and is the result of correcting for axial chromatic abberation in apochromatic objectives. This difference has traditionally been corrected in the eyepiece, and a manufacturer will typically design the same amount of correction into all their objectives so that all can be used with the same eyepiece. This practice has led to the advice against mixing eyepieces and objectives from different manufacturers.
(The Handbook of Optics, published by the Optical Society of America and McGraw Hill lists a number of eyepieces from a variety of manufacturers. The CDM for many of these eyepieces were listed, with all in the range of +0.3% to -1.5%. The microscopes in my laboratory used eyepieces which had a -1.4% correction, and the chromatic abberations were discernible at the periphery of the micrographs produced on those scopes if a correcting lens was not used.
Nikon introduced their "Chrome-Free" optics in 1976, and the Zeiss microscopes introduced in 1985 also corrected for CDM in the intermediate optics of the microscope. The objectives used in stereo microscopes do not exhibit CDM and do not need CDM corrections in the eyepiece. (Wild microscopes are an exception.)
Since one of the major uses for the MicroCam is for stereo microscopes, (often having no other photographic capability), and since the trend was toward correction before the eyepiece, we chose to incorporate in the MicroCam an eyepiece with no CDM. Therefore no fringing should be seen with chrome free optics, and with most stereo microscopes. (Some zoom systems do introduce some fringing.)
One response comment that you received had a number of complaints about the MicroCam system. The first concerned focus and resolution. Care with focusing is of course critical. It is important that the focusing crosshair be seen sharply before focusing the image. The MicroCam uses integral film, so exposure of the negative is through a clear polyester cover sheet. This makes the integral film sensitive to excess flare, so it is also important the substage iris of the microscope be adjusted to minimize flare and maximize contrast. (There is always the balance between contrast and resolution, and the condenser iris should be set at that point just before resolution is lost.) This comment may seem pedantic, and you might respond "of course", but the control of flare is particularly important with this film. The resolution of the color integral film, is ~ 10 line pairs per millimeter. This matches the resolution in the microscope image as governed by the resolution capabilities of most microscope objectives. Integral black and white films have a higher resolution, ~20 line pair per millimeter.
The second concerned color rendition. I have a number of commercially prepared, moderately stained slides, mainly H&E, some aniline blue and I have been satisfied with their color rendition. The MicroCam incorporates a light balancing filter approximating an 80B, which would be the appropriate filter for a tungsten halogen lamp at its rated voltage. Some adjustment of filtration may be necessary with the users own filters, to optimize color rendition for their light sources or for transmission characteristics of their microscopes.
Third, indeed, you do wind up with a print, and not a negative or a transparency. The print is self-developing outside the camera. I have used it with a Polaroid scanner, to get an electronic image for image analysis or incorporation in communications.
The fourth point mentioned that "Exposure varies with the specimen so many test shots are necessary." The exposure control measures the entire image area, and responds similarly to any other automatic camera system averaging a large area. The camera has electronic correction for reciprocity failure, both for speed and for color balance. The exposure adjustments are clear and easily accessible, for specimens with widely varying backgrounds.
I hope I have answered your question, and some of the other questions which have been raised. If I can be of further help, my email address is "mccanns-at-tiac.net".
Having retired from Polaroid, I no longer have any financial interest in this product. However, I am proud of this product and its role as a versatile instant camera with automatic exposure control providing low cost photomicrography capability.
Mary McCann McCann Imaging email: mccanns-at-tiac.net Tel: 617-484-7865 Fax: 617-484-2490
Nuria Cortadellas wrote: ============================================== } I have a big problem, osmium precipitates!! } does anyone can help me? ============================================== Are you sure it is "osmium" (probably OsO2 if it is of osmium composition) and not iron oxide contamination from corrosion from the tweezer tips? Vapor staining with osmium tetroxide does corrode the tips of the "normal" antimagnetic stainless steel types. And such corrosion product can migrate to a sample being supported on a TEM grid. While the chances are greater that the problem is indeed from the osmium tetroxide, that might not always be the case.
Disclaimer: Our firm, SPI Supplies offers alternatives to the "normal" antimagnetic stainless steel tweezers for holding grids so we have a vested interest in making this point.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
} } From: Nuria Cortadellas \ Internet: (nuriac-at-giga.sct.ub.es) } To: MICROSCOPY BB \ Internet: } (microscopy-at-sparc5.microscopy.com) } } Subject: Osmium precipitates } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I have a big problem, osmium precipitates!! } does anyone can help me? } Thanks in advance } Nuria Cortadellas } Department of Electron Microscopy } University of Barcelona } } }
-------------------------------------------------------------- Peling Fong Melville Senior Scientific Assistant Interdepartmental Facilities American Museum of Natural History Central Park West at 79th Street New York, NY 10024-5192 U.S.A. ****************************** E-mail: peling-at-amnh.org Work #: (212) 769-5469 FAX #: (212) 769-5495
After discussed dangers of the use of DMSO suggested by Wil Bigelow, I have to share with you the old knowledge that the same effect on small burns can be obtained by a simple use of the egg white. You remove the membrane lining the egg shell and put it on the burned area. If needed, you add more of these membranes. It stops pain immediately and prevents blister formation. The method is apparently known for centuries and around the world, as the egg white treatment was also described by Gabriel Garcia Marquez in "One Hundred Years of Solitude". I never experienced, nor heard about, any negative effect.
I have an older TEM with many extras for which a new home is needed. This is a Philips EM301 with both the high resolution stage and the goniometer stage. It is presently owned by a friend whos husband died before he could finish putting his lab together. The price is negotiable and the instrument, like most of the older Philips microscopes is quite servicable. If you have any interest or questions, please contact me via e-mail.
Thank you. ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ Alexander Greene Scientific Instrumentation Services, Inc. Number 499, Post Office Box 19400 Austin, Texas 78760 Phone: 512/282-5507 FAX 512/280-0702
REASONABLY PRICED ELECTRON MICROSCOPE REPAIR ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
I would like to add to Linda's recommendations that this precipitate phenomenon may be most common in EM but it is less obvious than lead precipitates. It was published (don't ask where) over ten years ago. It's a triangle which requires phosphate, GA and Os. If no free GA remains after thorough rinsing with buffer, the Os will not result in the precipitate. A rinse which would remove precipitate (prior to the sections irradiation by an electron beam) was also published, perhaps somebody else can post that, I do not remembers the detail. In the end cacodylate solved the precipitation problem and rinsing between GA and Os is not required. It also causes no precipitation with Ca (seawater) and results in better preservation. Pity is, because phosphate buffer is o.k. to drink, whereas cacodylate is an arsenic compound which is toxic and is a carcinogen. I believe reasonable facilities and good working habits can make it quite safe to use. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 350+ Links, MSDS ************************ http://www.proscitech.com.au
---------- } Reply to: RE} osmium precipitates } } Dear Nuria, } Sounds like your phosphate buffer interacting with the osmium may be the cause } of precipitates. We generally fix for resin embedding in cacodylate buffer. If } phosphate buffer is used for the fixative, we wash it away 3 x 10 minutes with } cacodylate buffer then proceed with osmication. The osmium is also never } diluted in phosphate buffer. We keep a stock solution of 4% made in water then } dilute to 2% with cacodylate buffer when ready to use. Hope this helps. } Linda Chicoine } Center for Cell Imaging } Dept. of Cell Biology } Yale University
Hi Jolanta,
My recipe is (and it is the recipe from Polysciences who sells all the LWs : For K4M it is : Crosslinker A : 3.6 ml ( or 2.7g) Monomer B : 25 ml ( or 17.3 g) Initiator C : 100mg (and NOT 20.1g)
They say : 1 weigh out, into a tared vial, the crosslinker and the monomer. Mix gently by one of the following methods for three to five minutes : =
-bubble a continuous stream of dry nitrogen gas into the mixture with a Pasteur pipette. The nitrogen stream will mix the resin, and at the same time it will prevent the incorporation of oxygen. -mix gently with a glass rod. -if the vial has a small cap or lid, slowly rock the covered vial from side to side, avoiding the formation of air bubbles or foaming. Add the initiator and continue mixing until the initiator is completely dissolved in the resin. The mixture is given for ultraviolet polymerization from -50=B0C to 0=B0C= =2E Above 0=B0C, the initiator C should be replaced by the same amount of benzoin ethylether.
IN ALL THE LWs MIXTURES, YOU ONLY USE 100 TO 150 MG OF INITIATOR
You might wish to ask EMS for their booklet on the use of lowicryl and their lowicryl letters. =
My recipe is (and it is the recipe from Polysciences who sells all the LWs : For K4M it is : Crosslinker A : 3.6 ml ( or 2.7g) Monomer B : 25 ml ( or 17.3 g) Initiator C : 100mg (and NOT 20.1g)
They say : 1 weigh out, into a tared vial, the crosslinker and the monomer. Mix gently by one of the following methods for three to five minutes : =
-bubble a continuous stream of dry nitrogen gas into the mixture with a Pasteur pipette. The nitrogen stream will mix the resin, and at the same time it will prevent the incorporation of oxygen. -mix gently with a glass rod. -if the vial has a small cap or lid, slowly rock the covered vial from side to side, avoiding the formation of air bubbles or foaming. Add the initiator and continue mixing until the initiator is completely dissolved in the resin. The mixture is given for ultraviolet polymerization from -50=B0C to 0=B0C= =2E Above 0=B0C, the initiator C should be replaced by the same amount of benzoin ethylether.
IN ALL THE LWs MIXTURES, YOU ONLY USE 100 TO 150 MG OF INITIATOR
You might wish to ask EMS for their booklet on the use of lowicryl and their lowicryl letters. =
} I desperately need to convert "old spectra files" from } old systems to the newer one Intel/Windows platform and from/to Edax {--} Link !
Dear Andrea, Here at Mektech we manufacture MS Windows based EDS system that connects to Link AN10000 pulse processor. It can also read Link AN10000 and eXL spectra. For more info visit our website at www.visionol.net/~mektech or contact as directly.
I have been asked to study electrodeposited copper microstructure. My goal is to determine the relationship among electrodeposition process operational variables (electrolyte concentration, impurities deposition, aditives, current intensity applied etc.), microstructure (texture, grain size...), and mechanical properties of the metal.
You are all welcome to visit 5 copper micrographs I have posted on the Internet and please fell free to make any comments on them:=20
http://metallography.com/marti.htm
Any ideas will be welcome. Any reference info on the specific subject of electrodeposited copper microstructure will also help me. Can you tell me where to find copper cathode micrographs?
Please reply to Juan Marti at: jmartip-at-cepade.es
The descriptuion of the pictures is as follow:
The 5 micrographs correspond to the same cathode sample and show different structures among which I am not able to determine which one is the real one. I hope that you may help me in the interpretation of these images:
- Bottom left picture (cobre4). Etched with HNO3 (25%) at 70=BAC during only 5 seconds. Longitudinal section of the cathode showing what seems to be the grain boundaries. Lens Objective: 50x.
- Upper left and right pictures (cobre1). Etched with HNO3 (25%) at 70=BAC during 15 seconds. Longitudinal section of the cathode. It apparently shows big irregular grains but I=92m not sure if the sample might be under etched, thus not showing the real microstructure. Lens Objective: 50x.
- Middle right picture (cobre3). Etched with HNO3 (25%) at 70=BAC. More etching time. Longitudinal section of the cathode showing much smaller grains. Grains also seem irregular. Lens Objective: 50x.
- Middle left picture (cobre2). Etched with HNO3 (25%) at 70=BAC. Transversa= l section of the cathode showing what appears to be large grains grown in the current flow direction. Lens Objective: 20x.
- Bottom right picture (cobre6). Shows detail of abnormal grain size. Lens Objective: 20x.
Among the first 3 micrographs (cobre4, cobre1 and cobre3) can you tell which one is the real pure copper microstructure?
Any ideas will be welcome. I would also appreciate any help on specific references to copper cathode microstructures descriptions and micrographs.= =20
Thanks in advance.
Please reply to Juan Marti at: jmartip-at-cepade.es
Micrographs web site at: http://metallography.com/marti.htm
Biological Physics. Postdoctoral position at the Department of Molecular Physiology and Biological Physics of the University of Virginia School of Medicine. The research projects in which the applicant is expected to play a major role involves electron energy loss spectroscopy and energy filtered scanning transmission electron microscopy. The position will include both development of software and instrumentation for achieving 2 to 3 nm spatial resolution compositional imaging and hands-on application of the method to significant biological problems. The laboratory has been engaged in NIH- supported research developing and applying analytical electron microscopy for 25 years through an interdisciplinary program based on the collaboration between physicists and biologists. Equipment available includes a 200kV electron microscope equipped with field emission gun (Philips CM200-FEG), a GATAN electron spectrometer adapted to our own CCD camera, a CM12 electron microscope and two energy dispersive X-ray detectors. Investigators in the program are also members of the Center for Structural Biology of the University of Virginia and have programs involving collaborations with the X-ray crystallography and atomic force microscopy groups and with molecular biologists and investigators engaged in other biological disciplines.
Candidates should have a Ph.D. in Physics, material science or engineering and be familiar with computer programming, instrumentation and, preferably, experience with electron microscopy and electron energy loss spectroscopy. Applications with biographical sketch, bibliography and the names of three references should be sent to: Dr. Andrew P. Somlyo, Department of Molecular Physiology and Biological Physics, University of Virginia, P.O. Box 10011, Charlottesville, VA 22906-0011, USA. The University of Virginia is an Equal Opportunity/Affirmative Action Employer.
I apologize for the false start. It's for the Philips EM201/300/301 series microscopes we need (NOT the 200 or 400, as I thought earlier). Again, used, simple, single tilt, is fine, if not bent. Key parameter is inexpensive.
DGCollins-at-lbl.gov (510) 486-7859, or DCollins 2841 Kinney Dr, Walnut Creek, CA 94595 (510)939-2006
There is a program (NTRANS) in the MSA Software library which runs on your PDP 11 computer and can do SOME of the work you request. The software is free, but you will have to get someone to compile it. Alternatively, you ask the manufacturers to supply to you a copy of their program which translates their data into the MSA/MAS Standard Spectral File Format. This should be even more effective, as each manufacturer only need to write a R/W subroutine for their format to/from the one standard. The advantage here is that the manufactures may already have the translator compiled and running on each of the platforms that you already have. The other option is DTSA which is a National Institute of Standards and Technology (NIST) computer program for XEDS , which has many translators built in. That program however must be purchased from NIST. (Disclaimer: I have no financial interests in DTSA).
If you want a Copy of NTRANS you can get it here;
The anonymous ftp server address is
Host: ftp.msa.microscopy.com UserId: anonymous Passwd: your email address
or you can go to the master ftp site
Host: ftp.amc.anl.gov UserId: anonymous Passwd: your email address
Go to the public directory, find the MMSLib Go to the XEDS directory Go to the NTRANS directory
All the files are in there...
Here is a copy of the on-line abstract file
Ntrans.abs.
Title :NTRANS Keywords :XEDS, EELS Computer :DEC VAX 11/730-785, DEC PDP 11/2-11/73 Operating System :VAXVMS, RT-11 Programming Language :Fortran IV Hardware Requirements :None Author(s) :Nestor J. Zaluzec Correspondence Address :Argonne Nat. Lab, Electron Microscopy Center,Bldg 212 :Materials Science Division, Argonne, Illinois 60439, Abstract:
NTRANS is a computer program which translates manufacturers XEDS and EELS spectral data into the EMMPDL data format. It utilies the RWEMMPDL subroutines contained in the EMMPDL. At present this version translates both EDAX and TRACOR-Northern and Link Systems data files. Examples of translated spectra can be found spectra can be found in the XEDS and EELS subdirectories of the EMMPDL. -------------------------------------------------------------------------------
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} I buffered the osmium solution with phosphate and cacodylate } buffer and the osmium precipitates appear in the sample
Residual gluaraldehyde is famous for causing Os04 precipitates. Although you say that you rinse 4-5x, try rinsing more to be sure that you are removing all GA. Also, if your specimen size is too large, the GA may not be diffusing out of the tissue entirely because of the great distance.
(Just my two cents worth),
Good luck. ______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 The College of New Jersey fax: (609) 771-2674 Trenton, NJ 08650-4700
Having spent several years wordering just how many different formats Link could possibly come up with for storing X-ray spectra, I was just a little bit enraged when they changed things AGAIN with the introduction of the ISIS system. The converters Nestor has pointed out won't work with that system.
I'd like to echo Nestor's suggestion to the manufacturers about them supplying converters and it seems that this forum could be a useful place to gather a weight of opinion. I know that there are many parameters that are sometimes stored along with the 'raw' data which would make the free translation of data formats somewhat perilous or difficult, but it ought to be possible for access to most of the data to be a darned sight easier than it is now.
Dr Simon Dumbill AEA Technology Tel: +44 1235 434245 220, Harwell Fax: +44 1235 435941 Didcot Email: Simon.Dumbill-at-aeat.co.uk Oxfordshire OX11 0RA UK
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Andre'
There is a program (NTRANS) in the MSA Software library which runs on your PDP 11 computer and can do SOME of the work you request. The software is free, but you will have to get someone to compile it. Alternatively, you ask the manufacturers to supply to you a copy of their program which translates their data into the MSA/MAS Standard Spectral File Format. This should be even more effective, as each manufacturer only need to write a R/W subroutine for their format to/from the one standard. The advantage here is that the manufactures may already have the translator compiled and running on each of the platforms that you already have. The other option is DTSA which is a National Institute of Standards and Technology (NIST) computer program for XEDS , which has many translators built in. That program however must be purchased from NIST. (Disclaimer: I have no financial interests in DTSA).
If you want a Copy of NTRANS you can get it here;
The anonymous ftp server address is
Host: ftp.msa.microscopy.com UserId: anonymous Passwd: your email address
or you can go to the master ftp site
Host: ftp.amc.anl.gov UserId: anonymous Passwd: your email address
Go to the public directory, find the MMSLib Go to the XEDS directory Go to the NTRANS directory
All the files are in there...
Here is a copy of the on-line abstract file
Ntrans.abs.
Title :NTRANS Keywords :XEDS, EELS Computer :DEC VAX 11/730-785, DEC PDP 11/2-11/73 Operating System :VAXVMS, RT-11 Programming Language :Fortran IV Hardware Requirements :None Author(s) :Nestor J. Zaluzec Correspondence Address :Argonne Nat. Lab, Electron Microscopy Center,Bldg 212 :Materials Science Division, Argonne, Illinois 60439, Abstract:
NTRANS is a computer program which translates manufacturers XEDS and EELS spectral data into the EMMPDL data format. It utilies the RWEMMPDL subroutines contained in the EMMPDL. At present this version translates both EDAX and TRACOR-Northern and Link Systems data files. Examples of translated spectra can be found spectra can be found in the XEDS and EELS subdirectories of the EMMPDL. -------------------------------------------------------------------------------
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
After read all the posted messages about Glutaraldehyde and Formaldehyde with their hazadous fumes, we Electron Microscopy Sciences want to remind you all that we have been introducing in our catalog, in the Safety Section, the LAB-AIR System - an Electronic Air Purifiers, which complied with OSHA Regulations and minimizes Occupation Exposure To Toxic Vapors. The System that destroys the odors and fumes, and doesn't just mask them. Electronic Air Purifiers produce a controlled level of Ozone (O3) electrically by converting molecules of Oxygen (O2) into molecules of Ozone (O3).. Ozone, sometimes called activated oxygen, as part of the process of returning to oxygen, casts off its extra atom. That extra atom combines with the molecule of the odor's source and thereby destroys the odor by oxidation. Once Ozone's extra atom is consumed fresh air is leftbehind which was created by a natural process. For instance: HCHO + O3 = HCOOH + O2 Formaldehyde Ozone Formic acid Oxygen HCOOH + O3 = CO2 + H2O + O2 Formic acid Ozone Carbon dioxide * Water* Oxygen* * All Harmless Gases. We are not intended to introduce our product on the site, but we thought this messages are helpfull to all of our Scientists and Technicians, whose is dealing with chemicals daily in theirs enclosed labs. For more information, please contacting us at 1 800 523 5874
Electronic microscopy by transmission (Zeiss EM109)
We performed more than 20,000 photographies on not-perfored rollfilm 70 mm as AGFA Scientia, AGFA Rapidoline, AGFA Aviortho, KODAK Kodalith. Unfortunately the production of all these rollfilms are now stopped , as well as Ortho AGFA films (sheet and roll 120/135).
Is anybody has suggestions about the replacement of these high specific products ?
In fairness to Oxford, it should be mentioned that the ISIS may have its own new file format for spectra, but it does read and write the earlier eX-L and AN10000 formats, as well as the EMSA/MAS format.
The x-ray analyser manuals for the AN10000 and the eX-L both contain detailed descriptions of the spectrum file formats in appendices. The MSDOS convert program, available on eX-L's and later AN10000's (those with 3.5" floppy's, I think) will copy the spectra to MS-DOS disks, from which a simple program will readily convert them to text files.
I have written such a program. It is available by anonymous FTP from IMAGES.MIT.EDU. There are several files there, but the readme files explain what is what. In addition to the conversion program above, there is a program to duplicate the MSDOS Convert program (by reading Genie/DEMON 3.5" disks on the PC) and a program to extract images from studies.
Hope this is useful.
Tony Garratt-Reed
**************************************************** **************************************************** ** ** ** Anthony J. Garratt-Reed ** ** Room 13-1027 ** ** Center for Materials Science and Engineering ** ** Massachusetts Institute of Technology ** ** 77 Massachusetts Avenue ** ** Cambridge, Massachsetts 02139-4307 ** ** U. S. A. ** ** ** ** Phone: 617-253-4622 ** ** Fax: 617-258-5286 or 617-258-6478 ** ** ** **************************************************** ****************************************************
Please be advised of the following position opening. Contact John May at the address below.
Bob Wise
} Date: Mon, 16 Jun 1997 10:55:14 -0500 } From: "John F. May" {shayala-at-fdldotnet.com} } Subject: Electron microscopy position } X-Sender: shayala-at-pop.fdldotnet.com } To: wise-at-vaxa.cis.uwosh.edu } Cc: jedunphy-at-mariancoll.edu } X-Mailer: Windows Eudora Light Version 1.5.4 (32) } } Dr. Wise, } } I would like to alert you to a FULL TIME faculty position in Biology } at Marian College in Fond du Lac for the 1997-98 school year. We want } someone to teach the following courses: } } Bi/PhS 331 Transmission Electron Microscopy (2 cr.) } Bi 100 Life Systems (lecture only) (3 cr.) } Bi 100 Life Systems lecture & lab (evening section) } (4 cr.) } Sci 101 Integrated Physical/Biological Science (3 cr.) } (team-taught with a Phsycal Science } faculty member) } OR } Bi 201 Anatomy & Physiology (4 cr.) } } } The Spring semester schedule would be similar, with Sci 102 and Bi 202 being } offered as the second semester of those courses. The faculty member would } be expected to teach the Biology portion of the integrated course. } } Our normal course load is 24 hours per year. } } Please share this announcement with anyone you feel would be interested or } colleagues who might know a potential applicant. Have interested } individuals contact me be e-mail or phone. } } Thank you. } } John F. May, Ph.D. } Biology Coordinator } Marian College } Fond du Lac, WI 54935 } Phone: 414-923-7646 } e-mail: shayala-at-fdldotnet.com } }
Ozone sounds good in theory. I'd certainly prefer it to formaldehyde, but is that a "Hobson's choice" (choosing between the lesser of two evils)? Can you comment on the effects of ozone on lung tissue?
} To all, } } After read all the posted messages about Glutaraldehyde and Formaldehyde with } their hazadous fumes, we Electron Microscopy Sciences want to remind you all } that we have been introducing in our catalog, in the Safety Section, the } LAB-AIR System - an Electronic Air Purifiers, which complied with OSHA } Regulations and minimizes Occupation Exposure To Toxic Vapors. The System } that destroys the odors and fumes, and doesn't just mask them. } Electronic Air Purifiers produce a controlled level of Ozone (O3) } electrically by converting molecules of Oxygen (O2) into molecules of Ozone } (O3).. Ozone, sometimes called activated oxygen, as part of the process of } returning to oxygen, casts off its extra atom. That extra atom combines with } the molecule of the odor's source and thereby destroys the odor by oxidation. } Once Ozone's extra atom is consumed fresh air is leftbehind which was created } by a natural process. } For instance: } HCHO + O3 = HCOOH + O2 } Formaldehyde Ozone Formic acid Oxygen } HCOOH + O3 = CO2 + H2O + O2 } Formic acid Ozone Carbon dioxide * Water* Oxygen* } * All Harmless Gases. } We are not intended to introduce our product on the site, but we thought this } messages are helpfull to all of our Scientists and Technicians, whose is } dealing with chemicals daily in theirs enclosed labs. } For more information, please contacting us at 1 800 523 5874 } } Bang Nguyen } Electron Microscopy Sciences } } Dr. M. Dennis Goode Phone (301) 405-6917 Department of Zoology Fax (301) 314-9358 University of Maryland e-mail goode-at-zool.umd.edu College Park MD 20742 ************************************************************* "If the Lord Almighty had consulted me before embarking upon the creation, I should have recommended something simpler." -Alphonso X of Castile, 15th Century
I am currently trying to get Link to provide a simple X-Y output (channel- counts) of their spectra in tab-separated variable format from their ISIS system. That way I could easily import the spectra into Kaleidagraph or Excel. I don't care about the headers or other information; I can get that from the original data. They are "working on it", but I haven't heard anything from them for a month or so.
I'm not a programmer, but is it that hard to make an output like that? Since the spectra are plotted within their program it seems that those raw data in X-Y format MUST exist somewhere.
If they come up with something, I'll let people know.
Cheers,
John Vetrano Pacific Northwest National Laboratory _______________________________________________________________________________
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Having spent several years wordering just how many different formats Link could possibly come up with for storing X-ray spectra, I was just a little bit enraged when they changed things AGAIN with the introduction of the ISIS system. The converters Nestor has pointed out won't work with that system.
I'd like to echo Nestor's suggestion to the manufacturers about them supplying converters and it seems that this forum could be a useful place to gather a weight of opinion. I know that there are many parameters that are sometimes stored along with the 'raw' data which would make the free translation of data formats somewhat perilous or difficult, but it ought to be possible for access to most of the data to be a darned sight easier than it is now.
Dr Simon Dumbill AEA Technology Tel: +44 1235 434245 220, Harwell Fax: +44 1235 435941 Didcot Email: Simon.Dumbill-at-aeat.co.uk Oxfordshire OX11 0RA UK
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Message-Id: {1.5.4.32.19970616180604.0068f0ac-at-pop.fast.net} X-Sender: goldmrkr-at-pop.fast.net (Unverified) X-Mailer: Windows Eudora Light Version 1.5.4 (32) Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Colleagues -
A friend of mine is interested to sell his microscope, so I said I would put it on the server since he is not a member. Please contact me if you have an interest and I'll pass the word.
Thanks and regards, Don Cox
---------------------------------
CARL ZEISS RESEARCH MICROSCOPE (No model # that I can find---but it is their standard research microscope body that they carried for years. Probably bought in the mid 70's. } } Trinocular head with beam splitter. } } Condenser (phase and darkfield) } } Seven (7) zeiss lenses: } 16X /0.40 Neofluar, (phase) } } 40X /0.65 Plan } } 40X /0.65 Plan (phase) } } 40X /1.0 Oil (APO) Iris Diap. (0.6-1.0) } } 100X /1.25 Oil Iris Diap. (1.25-0.8)
Hello, We are starting a histochemical EM study on acetylcholinesterase activity in brain tissue. We are searching for a method of quantifying the levels of AChE activity. I would appreciate any information from an experienced researcher, and any appropriate references.
Recently, I am having problems when sectioning my blocks. I work on=20 animal tissue, a sea pansy which is a coelenterate. I only use the polype.
MY PROBLEM: when I section, I only get the resin and where my tissu should= =20 be I have a hole!! I never had this problem. Could it be a deshydratation too long (10 min. for each ethanol, 2*10 for the 100% and= =20 2*15 for oxyde propylene.. Or residual oxide propylene..Or could it be the= =20 inclusion in agarose. My resin is hard but when I come close to the tissue it is smooth.
Is it possible to recuperate these sections? and how
My PROTOCOLE BRIEFLY: I fix my tissue in glutaraldheyde or=20 paraformaldheyde 4% (various=20 fixation I use. ) After fixing for 3 or 4 hours, I embed the tissue in an= =20 agarose solution for 24 hours and I then section those agarose blocks=20 with a vibratome. Those sections of a thickness varying between 50 to=20 200 micrometers are incubated in 1% OsO4 for 2 hours. I then dehydrate in= =20 ethanol and embed in epon and araldite for 24 hours at 60 degrees. =09=09 =20 I hope that someone can help=20 thank you.
Natacha Benrimoh Universt=E9 de Montr=E9al benrimon-at-ere.Umontr=E9al.CA (514) 343-6111 poste 1052.
To sidetrack a bit, Hobson's choice actually means "no choice", there's only one recourse and that's it!!! Hope you don't mind a bit of ribbing, Dennis! :-) As for effects of ozone on lung tissue, I'm no chemist but as I understand it, it's a free radical and nothing good comes out of mixing those radicals with living tissue....that's why we have all those quacks toting beta-carotenes and Vitamin C & E etc. Well, that's my penny's worth of comments! :-)
Meng
On Mon, 16 Jun 1997, Dennis Goode wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Bang, } } Ozone sounds good in theory. I'd certainly prefer it to } formaldehyde, but is that a "Hobson's choice" (choosing between the } lesser of two evils)? Can you comment on the effects of ozone on } lung tissue? } } -Dennis } } } Date: Mon, 16 Jun 1997 07:46:30 -0400 (EDT) } } From: BNguyen260-at-aol.com } } To: goode-at-zool.umd.edu, Microscopy-at-sparc5.microscopy.com, } } richard.lander-at-stonebow.otago.ac.nz, KPR-at-wpo.nerc.ac.uk } } Subject: Re: Glutaraldehyde: safe limits -Reply } } } To all, } } } } After read all the posted messages about Glutaraldehyde and Formaldehyde with } } their hazadous fumes, we Electron Microscopy Sciences want to remind you all } } that we have been introducing in our catalog, in the Safety Section, the } } LAB-AIR System - an Electronic Air Purifiers, which complied with OSHA } } Regulations and minimizes Occupation Exposure To Toxic Vapors. The System } } that destroys the odors and fumes, and doesn't just mask them. } } Electronic Air Purifiers produce a controlled level of Ozone (O3) } } electrically by converting molecules of Oxygen (O2) into molecules of Ozone } } (O3).. Ozone, sometimes called activated oxygen, as part of the process of } } returning to oxygen, casts off its extra atom. That extra atom combines with } } the molecule of the odor's source and thereby destroys the odor by oxidation. } } Once Ozone's extra atom is consumed fresh air is leftbehind which was created } } by a natural process. } } For instance: } } HCHO + O3 = HCOOH + O2 } } Formaldehyde Ozone Formic acid Oxygen } } HCOOH + O3 = CO2 + H2O + O2 } } Formic acid Ozone Carbon dioxide * Water* Oxygen* } } * All Harmless Gases. } } We are not intended to introduce our product on the site, but we thought this } } messages are helpfull to all of our Scientists and Technicians, whose is } } dealing with chemicals daily in theirs enclosed labs. } } For more information, please contacting us at 1 800 523 5874 } } } } Bang Nguyen } } Electron Microscopy Sciences } } } } } Dr. M. Dennis Goode Phone (301) 405-6917 } Department of Zoology Fax (301) 314-9358 } University of Maryland e-mail goode-at-zool.umd.edu } College Park MD 20742 } ************************************************************* } "If the Lord Almighty had consulted me before embarking upon the } creation, I should have recommended something simpler." } -Alphonso X of Castile, 15th Century }
Dear All, I keep a Aloe Vera plant on my windowsill. If I get a burn, I just snip off a leaf and squeeze out the jelly onto the burn. It forms a protective skin and cools the hurt. Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
I spoke to Agar Scientific yesterday - they are our normal supply source for film. They are trying to obtain another film for me to try, but no details as yet.
We have used unperforated since 1967 (1969 personally) on Philips microscopes. We found many years ago that if the negatives were well focused and also in the darkroom then no-one could differentiate between prints from plates (as then) and 35mm film on 20 x 25 cm paper.
Not a bad idea. Aloe Vera has been used in the Tropics by many for a number of ailments. Good Ole Folk Medicine.
Leo
On Mon, 16 Jun 1997, Mary Mager wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear All, } I keep a Aloe Vera plant on my windowsill. If I get a burn, I just snip off } a leaf and squeeze out the jelly onto the burn. It forms a protective skin } and cools the hurt. } Regards, } Mary } } Mary Mager } Electron Microscopist } Metals and Materials Eng., UBC } 6350 Stores Rd. } Vancouver, B.C. V6T 1Z4 } CANADA } tel:604-822-5648, fax:604-822-3619 } e-mail: mager-at-unixg.ubc.ca } }
Hello everybody! Many thanks to all of you who have replied to my question about TEM film. Here, however, is a reply I got privately, which I am going to try out. Youall might like to try it, too, but I cannot yet vouch for any results.
* * We still run 35mm film in our EM Unit for both our Philips TEM's * (CM10 and 201c) as well as our Cambridge 250 SEM. * * However; on the TEM's we had the same problem you are now * experiencing. Changing to plates, I think is a step backwards. We use * to use Kodak FGP but we that became unavailable...we switched to AGFA * COPEX Pet 10 which we found as good. Perfect in the sense that it is * as sensitive, and not perforated. * } Does it have to loaded in the dark, or will red light do? (much more } convenient). * * A RED SAFELIGHT IS ALL WE USE *
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
} On the subject of LN2, does anyone know the correct first aid treatment } for a burn from this substance? } } Normally, one would use cold water to cool a (heat) burn and prevent } further tissue damage. But that seems inappropriate somehow...Hot water? } Dear Anthony, To quote from p. 41 of the Electron Microscopy Safety Handbook, 2nd Ed.: "Cryogens cause burns similar to frostbite and should be treated by warming of the affected part to body temperature." Yours, Bill Tivol
Please accept my apologies if I'm going over a well traveled path but I can't seem to find the answer to my question in my records from the list, namely: what are some of the best cameras to hook up to a TEM so that we can digitize images? We have a JEOL 100CX fitted with a YAG crystal from Fullam, a Macintosh 8500, and NIH Image for our software program. I imagine we would also end up purchasing a frame grabber such as one of the SCION boards.
But meanwhile what about a camera? I know there are quite a few out there varying widely in price and capabilities and I guess I'm wondering if anyone could help me with some feedback? I'm anticipating being able to spend between $10K and (hope hope) $20K. Any and all information will be greatly appreciated!
In 1992, we studied corneal endothelial cell damage as a result of incidental contact during ocular surgery. Although we were not concerned with the epithelium or Bowman's layer, we found a stain that worked quite well with the endothelium; Trypan Blue at a concentration of 0.2%. If it helps, please refer to my article in the 1992 EMSA proceedings, Part II, p. 1106:
"Evaluation of the Biocompatibility of Polymer Surface Modifications with the Corneal Endothelium", R. Citron, B. Tunberg, A. Yamada.
Regards,
Bob ************************* Bob Citron Chiron Vision Claremont, CA 91711 (909)399-1311 Bob_Citron-at-cc.chiron.com *************************
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am working with the human cornea and am exploring options for tissue preparation techniques and staining procedures for LM. Sections of 0.5-1.0 microns will be used to distinguish between the epithelium and underlying Bowman's layer. I need to find the optimal embedding medium as well as staining procedure.
EDAX Offers a conversion program for the PV9900 called PVconvert. The DX-4 system allows you an option to save spectra as a .csv file for spreadsheet use.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am currently trying to get Link to provide a simple X-Y output (channel- counts) of their spectra in tab-separated variable format from their ISIS system. That way I could easily import the spectra into Kaleidagraph or Excel. I don't care about the headers or other information; I can get that from the original data. They are "working on it", but I haven't heard anything from them for a month or so.
I'm not a programmer, but is it that hard to make an output like that? Since the spectra are plotted within their program it seems that those raw data in X-Y format MUST exist somewhere.
If they come up with something, I'll let people know.
Cheers,
John Vetrano Pacific Northwest National Laboratory _______________________________________________________________________________
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Having spent several years wordering just how many different formats Link could possibly come up with for storing X-ray spectra, I was just a little bit enraged when they changed things AGAIN with the introduction of the ISIS system. The converters Nestor has pointed out won't work with that system.
I'd like to echo Nestor's suggestion to the manufacturers about them supplying converters and it seems that this forum could be a useful place to gather a weight of opinion. I know that there are many parameters that are sometimes stored along with the 'raw' data which would make the free translation of data formats somewhat perilous or difficult, but it ought to be possible for access to most of the data to be a darned sight easier than it is now.
Dr Simon Dumbill AEA Technology Tel: +44 1235 434245 220, Harwell Fax: +44 1235 435941 Didcot Email: Simon.Dumbill-at-aeat.co.uk Oxfordshire OX11 0RA UK
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
What are the people at Marion College thinking? They want someone to teach electron microscopy and three other courses in one semester? I have taught a 4-hour course in electron microscopy for 10 years. The first 7 I was on my own and was spending better than 50 hours a week just with that course. The last three years I have had an assistant and the two of us stay very busy. If electron microscopy is taught hands-on, it is a labor intensive course for both the students and the teachers, and the only way it can be a useful and meaningful course is to be hands-on. That is what it is all about. One can read all the theory in the world, but nothing substitutes for sitting in front of an ultramicrotome for a few hours a day and putting both hands on all those neat controls on the electron microscope.
You are nearly certainly experiencing problems with tissue falling out of the section because of suboptimal infiltration. You probably have a very hard item which is difficult to infiltrate. Epon-Araldite combinations are very viscous, but the adhesive qualities of Araldite make it a good choice in these cases. Try infiltration with propylene oxide and resin 2:1 for one hour, 1:1 for 2 hours, 1:3 for 3 hours. Then pure resin for one hour. Then pure resin (new tube) for overnight. In the AM change resin again and rotate for another 2 hours. Your specimen vials must be in motion the entire time that infiltration is taking place. If your problem is not solved this way, please call or E-mail me. There may be other influences at work here. I assume that you have a sharp diamond knife at the correct angle, etc. Bye, Hildy Crowley hcrowley-at-DU.edu
I agree 200% with you on this Joyce. It's about time some of these administrators are set straight regarding EM. True it is a tool, but a very labor intensive one both to learn and to use. I just rubs me the wrong way to hear of all the downsizing and putting undertrained, undereducated people in charge of EM labs. I can't help but howl when I see an EM position requiring extensive knowledge and training in many sophisticated techniques for a $22,000 annual salary.
my $00.02 worth ed
Edward J. Basgall, PhD The Pennsylvania State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618 } } What are the people at Marion College thinking? They want someone to } teach electron microscopy and three other courses in one semester? } I have taught a 4-hour course in electron microscopy for 10 years. The } first 7 I was on my own and was spending better than 50 hours a week } just with that course. The last three years I have had an assistant and } the two of us stay very busy. } If electron microscopy is taught hands-on, it is a labor intensive } course for both the students and the teachers, and the only way it can } be a useful and meaningful course is to be hands-on. That is what it is } all about. One can read all the theory in the world, but nothing } substitutes for sitting in front of an ultramicrotome for a few hours a } day and putting both hands on all those neat controls on the electron } microscope.
Some months ago I asked for suggestions regarding the best way to handle mechanical pump oil vapors. We had a problem with pumps working in a room with people and computers. I received many helpful replies and thought I should share our experience.
First, it is the unanimous opinion of everyone who responded and with all those I checked with that exhausts from mechanical pumps and people do not mix. For reasons of health, fire safety, cleanliness and liability, pump exhaust should be controlled.
Second, many filters designed to stop oil mist from pumps may not be sufficient to capture all the vapors and/or they may need more frequent changing to be effective, ie if you smell oil vapor, filter or not, you need to fix it.
Third, the best solution to the problem is to vent the pumps to the outside or at least to the exhaust ventilation of the building.
So, here is how we tried to solve our problem of controlling pump exhaust. I proposed that the pump exhausts be connected to the building ventilation system. I did not get too far with this plan. I got a lot of resistance, mostly based on the expense involved with ducting (it is about 50' to the nearest fume hood) and rebalancing the air circulation system of the entire 5 floor building. The story was that it would take an incredibly long time to get anything like that done because it would involve getting architects and engineers to plan and spec the project etc.
I tried to enlist the help of our health& safety office but they couldn't do much because there seem to be no guidelines to use to determine if we were violating health standards. It was sort of a Catch-22, they agreed oil mist was probably not healthy, but they could not use their leverage to demand building modifications because there were no standards to enforce.
So, it was back to the drawing board for a solution. Several replies suggested using PVC pipe to make a pathway for the vapors either to the outside or to the nearest fume hood. I was already to try that when the campus fire dept. vetoed the use of PVC pipe for any kind of pump exhaust.
By now I had made good friends with one of the campus plumbers who was trying to help me with the job. He found an acceptable flame resistant hose and a large wall mounted filter that should do the job. The filter is supposed to remove all oil vapors and it has a pressure gauge to indicate when the filter should be changed. Only time will tell if this is a good solution to our problem, so far, so good.
Correcting the problem of mechanical pumps discharging oil mist into the air turned out to be more of a problem than I had anticipated. We are all breathing a lot easier in our lab now and I would encourage everyone to at least check their pump exhaust situation to make sure that it is adequately controlled.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
COMPUTER INTERACTIVE HIGH-RESOLUTION TRANSMISSION ELECTRON MICROSCOPY
N.C.E.M, LBNL, Berkeley, California
The National Center for Electron Microscopy announces its fourth ANNUAL SUMMER SCHOOL on COMPUTER INTERACTIVE HIGH-RESOLUTION TRANSMISSION ELECTRON MICROSCOPY, including Image Acquisition, Image Processing and Image Simulation, to be held at the National Center for Electron Microscopy during the week of August 25-29, 1997.
The aim of the School is to train participants in the techniques of computer-assisted high-resolution electron microscope image acquisition and image interpretation, including remote-control microscopy. Participants will learn general principles and apply them to specific cases. Participants will be taught the use of computers to obtain images on NCEM microscopes, followed by training in the use of application programs for image interpretation by image processing and image simulation. Participants wanting to apply school techniques to their own projects will be encouraged to extend their visit to NCEM into the next week -- note that this requires a proposal be submitted with advance notice sufficient for project approval.
For more information, please see - http://ncem.lbl.gov/NCEM/workshops.html
:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:. Michael A. O'Keefe, Deputy Head National Center for Electron Microscopy Lawrence Berkeley National Laboratory University of California Berkeley, California 94720 tel: (510) 486-4610 fax: (510) 486-5888 email: maok-at-lbl.gov http://ncem.lbl.gov/ :.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.
Request for Marine phytoplankton preparation protocol for scanning electron microscopy.
I am working on the structure of phytoplankton (Marine Chlorella, Isochrysis, Tetraselmis, Chaetocerus) before and after cryopreservation.
At present I am using this protocol 1. a. centrifuge the phytoplankton samples b. add ammonium formate to remove salts crystal. c. fix in 4% buffered gluteraldehyde for 12 to 24 hrs at 4 =B0C
2. a. washing with distill water for 3 changes of 10 minutes =09 each.
3. a. Fix in 1% buffered Osmium tetroxide for 2 hrs at 4 =B0C
4. a. Rinse in distill water for 3 changes of 10 minutes each.
5. Dehydration in a series of acetone a. 35% 5 minutes b. 50% 5 minutes c. 75% 5 minutes d. 95% 5 minutes e. absolute acetone 15 minutes, 3 changes
6. Viewing under electron microscope (JEOL 6400) The cell deformed/shrunken
Can you suggest other protocol for marine phytoplankton.=20
Your suggestion is very much appreciated.
Thank you.
Hishamuddin Omar Department of Biology Faculty of Science and Environmental Studies Universiti Putra Malaysia Malaysia
Hi all, I am after some advice on the care of the FE tip and vacuum on a Hitachi 4500.
1. How often should one bake out? The manual recommends baking out when the vacuum deteriorates. After about 8 months operation ours is better than it was to start with - IP1&2 off-scale, IP3 at 7x10-7 Pa. On the other hand many people seem to recommend baking at fairly short intervals "whether it needs it or not". We are inclined to a non-interventionist approach but are getting a bit nervous...any advice?
2. What should the flash current intensity be? Ours started at around 15-20 (and we sometimes flashed twice to get a reading in the high twenties) but has steadily crept up and is now in the high forties. Is this good, bad or indifferent? Is it perhaps related to question 1? If it gets too high does it wreck the tip? We are generally flashing once or twice a day.
cheers Sally ---------------------------------------------------------------------- Sally Stowe |Email: stowe-at-rsbs.anu.edu.au Facility Coordinator |Post: ANU Electron Microscopy Unit |ANUEMU (RSBS) Ph 61 6 249 2743 |Australian National Univ. FAX 61 6 249 4891 |Canberra, http://online.anu.edu.au/EMU/home.htm |AUSTRALIA 0200
It seems like you should't worry so much...the Hitachi FE-SEMs typically run many years under your conditions before tip replacement. Our experience is with a 10-year old S-800 and a 5-year old S-4500. The first emitter on the S-800 lasted 62 months, and the second is still working perfectly 61 months later. The first S-4500 emitter is, interestingly, 61 months old at present, and it still has the same flash characteristics as it started with...the flash current is in the mid-forty range. It is typically flashed once a day, or perhaps twice if the operation extends into the evening. I am not aware of any bake-outs of the gun except for the extensive (~10 day?)initial bake-out upon installation. Anecdotally, I have heard of Hitachi FE-SEMs whose emitters lasted in the 8 year range...perhaps you will have responses from operators of some of those instruments also with their experiences.
Larry
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
My E-Mail system has crashed a couple of times lately, and I believe that I have lost some messages. First, based on OHSS (Occupational Health and Safety Standards) provides that no employee should be in an environment where the ozone level on average is more than 0.1 parts-per-million for more than 40 hours per week or more than average of 0.3 parts-per-million for more than 15 minutes at any one time. ( Because of the limitation of the site, we can not display a chart , which is shows the Human Tolerance for Ozone). The LAB AIR Units are designed to produce the ozone level less than the limitation, and to operate within the OHSS guidelines. You should know that even with strong odor, the amount of ozone required to mask them out is only approximetely 0.04ppm, medium odor approx. 0.03ppm and light odor approx. 0.02ppm. Secondly, You are not breathing ozone air, you're breathing normal air, the Lab-Air turns on only when needed and ozone air is produced by the lab air just enough to mask (oxidizing) the odor sources. For instance, you're drinking water, not drinking chlorine, but in the water that you are drinking has some amount of chlorine, which is used to remove bacterias. When you do an embedding mixture, for instance !00ml Araldite-Epon mixture, you are using only maximum 1.5% of DMP-30 (1.5ml) to make the whole 100ml ot that mixture turn into a solid plastic block. Back to ozone air, you need just a small amount of O3 to oxidize the unwanted odor or unwanted chemicals which presence inside the room. Each Lab-Air has the timer to set the length of time which you want the Lab-Air to work, as well as setting for ozone to control the ozone air output, but the ozone output never exceeds the limit which is the Human Tolerance for Ozone. Thirdly, the O3 is unstable, which means it has a very short life, by its very nature Ozone will revert to oxygen within a short period of time.
Request for Marine phytoplankton preparation protocol for scanning electron microscopy.
I am working on the structure of phytoplankton (Marine Chlorella, Isochrysis, Tetraselmis, Chaetocerus) before and after cryopreservation.
At present I am using this protocol 1. a. centrifuge the phytoplankton samples b. add ammonium formate to remove salts crystal. c. fix in 4% buffered gluteraldehyde for 12 to 24 hrs at 4 =83C
2. a. washing with distill water for 3 changes of 10 minutes each.
3. a. Fix in 1% buffered Osmium tetroxide for 2 hrs at 4 =83C
4. a. Rinse in distill water for 3 changes of 10 minutes each.
5. Dehydration in a series of acetone a. 35% 5 minutes b. 50% 5 minutes c. 75% 5 minutes d. 95% 5 minutes e. absolute acetone 15 minutes, 3 changes
6. Viewing under electron microscope (JEOL 6400) The cell deformed/shrunken
Can you suggest other protocol for marine phytoplankton.
Your suggestion is very much appreciated.
Thank you.
Hishamuddin Omar Department of Biology =46aculty of Science and Environmental Studies Universiti Putra Malaysia Malaysia
Despite my occasional flippant remarks on various aspects of life in the Microscopy lane, sometimes about safety (being a Local Safety Advisor), I do take some aspects very seriously.
I came across a web page today which I feel is worthy of attention by those involved with or responsible for histotechnologists. It is about Repetitive Motion Disorder, Carpal Tunnel Syndrome etc. concerning the the use in particular of rotary microtomes.
I cannot believe (at present) that the reported statistical data relates only to this field, but it bears thinking about. As a sufferer, induced by home computer use, I can say that seious cases do not go away. Even typing this is a regular reminder tat it stays with you for a long time. It doesn't help typing accuracy either!
In our organisation (a nationally spread research council) there were 34 cases of RSI among computer users last year, among about 2,000 employees. This included one Laboratory Director! In this lab. in the last couple of weeks, a researcher has had a couple of weeks off with a suspected connection to pc use. We also have a an on-going case who is primarily a (light) microscopy user who records data simultaneously. The problem is such that our organisation recently called all Local Safety Advisors to head office for a special seminar on health and safety aspects of personal computers.
The web page referred to above is at:
http://www.hbu.de/rmd.htm#Definition I repeat - htm#Definition (I don't know what it means). This is part of Leica's web site.
I just got this request from a non-microscopist collegue, and I know there are people here better qualified to help him than I am. This would be for light microscopy. I have sent him some information and URLs, but more detailed ideas from experts would be appreciated. He is in Tasmania, so Australian product sources would be particularly useful.
} Do you know anything about using digital image "slices" to reconstruct 3-D } (kinda) images? There was an article in TREE last issue, and I had some } info on software from a company called Vaytek, but I'd like to know what is } needed to set up the microscope. } Alastair Richardson } alastair.richardson-at-zoo.utas.edu.au
Thanks.
Phil
} Sic Hoc Legere Scis Nimium Eruditionis Habes { Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217) 355-1143 oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
I agree with Ed and Joyce. I am in the process of finishing my PhD and I was curious about the job. I have over 15 years experience in EM and have assistant taught several other types of classes, so this job sounds like something I might have enjoyed...EXCEPT for the number of classes they wanted taught in one semester. My first thought was my husband would never see me again! The part about people wanting EM experts for minimum wage is also true. I used to supervise an EM research lab were I was responsible for ensuring grants and projects got done well and on time. My salary was equal to the secretarys'. They went home at 4:30 and I didn't.
Masters students in the graduate program here at UT Dallas have asked me about job opportunities in biological EM and after I tell them the salary they can expect, they usually loose interest in the area.
If my invert. zoo memory is correct the polyp form you speak of is mostly a gelatinous mass. If so a med. to softer resin would match your tissue, since a too hard resin compared to the tissue would also develop a falling out condition. The dehydration and PO times sound nominal but you could go an extra 10 min in 100% ETOH and 3 x10 min PO since the gelatin and your tissue does tend to retain water. Ms Crowley's suggestion of extended infiltration might also help. You shouldn't have trouble infiltrating into 200 micron (.2MM) thick tissue though. The type and amount of accelerator you use is not listed and that will have a dramatic effect on cutting quality. I have done several experiments with vibratome sections of chick embryos embedded in gelatin or egg yolk matrix using your technique and they came out fine, so it's just some fine tuning that your missing.
****************************snip************************** At present I am using this protocol 1. a. centrifuge the phytoplankton samples b. add ammonium formate to remove salts crystal= . c. fix in 4% buffered gluteraldehyde for 12 to 24 hrs at 4 =B0C
2. a. washing with distill water for 3 changes of 10 minutes each.
3. a. Fix in 1% buffered Osmium tetroxide for 2 hrs at 4 =B0C
4. a. Rinse in distill water for 3 changes of 10 minutes each.
5. Dehydration in a series of acetone a. 35% 5 minutes b. 50% 5 minutes c. 75% 5 minutes d. 95% 5 minutes e. absolute acetone 15 minutes, 3 change= s
6. Viewing under electron microscope (JEOL 6400) The cell deformed/shrunken ****************************end of snip*************** A few questions and suggestions to think about: Ammonium formate? buffered? pH? temperature? Temperature and pH- Try to keep it at or near that of the normal phytoplankton environment. Buffer osmolality - It should be nearly iso-osmotic to that of the organism or environment. Rinsing - It is generally not recommended to switch from buffer to water to buffer again. (buffered glut to dist water to buffered osmium) I would rinse in buffer and fix in buffered osmium then wash in dist water or use osmium in dist water followed by a dist water rinse. Dehydration - acetone may be harsher than ethanol. A continuous gentle gradient from 10% to 100% is preferred over a stepwise one. Drying method no mentioned? CPD? I would try HMDS and CPD in separate but identical preps. =46rom 100% ethanol to 1:1 ethanol:HMDS 10 min. to 100% HMDS (2-3x, 10 min.) then gently air dry over absorbent material (CaSO4 or silica gel). A gentle vacuum may be applied, using a water aspirator set-up.
good luck
ed
Edward J. Basgall, PhD The Pennsylvania State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618
****************************snip************************** At present I am using this protocol 1. a. centrifuge the phytoplankton samples b. add ammonium formate to remove salts crystal= . c. fix in 4% buffered gluteraldehyde for 12 to 24 hrs at 4 =B0C
2. a. washing with distill water for 3 changes of 10 minutes each.
3. a. Fix in 1% buffered Osmium tetroxide for 2 hrs at 4 =B0C
4. a. Rinse in distill water for 3 changes of 10 minutes each.
5. Dehydration in a series of acetone a. 35% 5 minutes b. 50% 5 minutes c. 75% 5 minutes d. 95% 5 minutes e. absolute acetone 15 minutes, 3 change= s
6. Viewing under electron microscope (JEOL 6400) The cell deformed/shrunken ****************************end of snip*************** A few questions and suggestions to think about: Ammonium formate? buffered? pH? temperature? Temperature and pH- Try to keep it at or near that of the normal phytoplankton environment. Buffer osmolality - It should be nearly iso-osmotic to that of the organism or environment. Rinsing - It is generally not recommended to switch from buffer to water to buffer again. (buffered glut to dist water to buffered osmium) I would rinse in buffer and fix in buffered osmium then wash in dist water or use osmium in dist water followed by a dist water rinse. Dehydration - acetone may be harsher than ethanol. A continuous gentle gradient from 10% to 100% is preferred over a stepwise one. Drying method no mentioned? CPD? I would try HMDS and CPD in separate but identical preps. =46rom 100% ethanol to 1:1 ethanol:HMDS 10 min. to 100% HMDS (2-3x, 10 min.) then gently air dry over absorbent material (CaSO4 or silica gel). A gentle vacuum may be applied, using a water aspirator set-up.
good luck
ed
Edward J. Basgall, PhD The Pennsylvania State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618
} Listers, } } I just got this request from a non-microscopist collegue, } and I know there are people here better qualified to help } him than I am. This would be for light microscopy. I have } sent him some information and URLs, but more detailed } ideas from experts would be appreciated. He is in } Tasmania, so Australian product sources would be } particularly useful. } } } Do you know anything about using digital image "slices" } } to reconstruct 3-D (kinda) images? There was an article } } in TREE last issue, and I had some info on software from } } a company called Vaytek, but I'd like to know what is } } needed to set up the microscope. Alastair Richardson } } alastair.richardson-at-zoo.utas.edu.au } } Thanks. } Phil
Dear Phil and Alastair, Sounds like you are talking about confocal microscopy. There are several light microscope companies offering this setup. Zeiss and Nikon to name just two. Breifly, only light from the plane of focus is collected. A stepping motor changes the focus a predetermined amount. The images are digitally collected and stored. Software then takes the images and stacks them to produce a 3D rendering. Depending upon the software it is possible to do many types of measurements, rotate the image, "fly" into it, examine each "slice" independtly. Learning how to use the software to get the most out of it is the hardest part. There is a listserver for confocal microscopy like this one but I don't know where it is. Hope this helps.
Gregory Rudomen University Microscopy Imaging Center S.U.N.Y. Stony Brook Greg-at-UMIC.SUNYSB.EDU 516-444-3126 *The opinions expressed above are my own and are not necessarily shared by the Microscopy Center*
I should add, please reply directly to Alastair at his email address given. } Listers, } } I just got this request from a non-microscopist collegue, and I know there } are people here better qualified to help him than I am. This would be for } light microscopy. I have sent him some information and URLs, but more } detailed ideas from experts would be appreciated. He is in Tasmania, so } Australian product sources would be particularly useful. } } } Do you know anything about using digital image "slices" to reconstruct 3-D } } (kinda) images? There was an article in TREE last issue, and I had some } } info on software from a company called Vaytek, but I'd like to know what is } } needed to set up the microscope. } } Alastair Richardson } } alastair.richardson-at-zoo.utas.edu.au ^^^^^^^^^^^^^^^^^^^^^^^^^^ } } Thanks. } } Phil
} Sic Hoc Legere Scis Nimium Eruditionis Habes { Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217) 355-1143 oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
} The carbon composite grid, while conductive, gives high Bremsstrahlung } background radiation reducing experimental sensitivites.
Since brehmsstrahlung production goes up strongly with Z, higher production from these grids can only be due to the greater amount of material in the path of the beam. If the total mass of the grid can be reduced to the same level as that for other grid materials, brehmsstrahlung would be quite low. Yours, Bill Tivol
We would be very interested in any recommendations and/or possible suppliers for the following:
1. An automatic negative processor which would handle black and white negatives that are 3.25 X 4 inches in size.
2. An automatic tissue processor to be used in processing for epon embedding.
Any experiences, good or bad, with similar equipment would be much appreciated.
I am also trying to contact Philip Slackman who originally contacted me after reading a message I left on this listserver. If you read this, Philip, please contact me again!
Pat Hales Dept. of Anatomy & Cell Biology McGill University hales-at-hippo.medcor.mcgill.ca
Sally, We have a S4000 which is a little older than yours, but should behave about the same. Firstly, I only bake out the vacuum after the power has been out for more than four hours, this happens about every six weeks in Utah. If the power would stay on longer I would bake less frequently.
Secondly, If you wish to extend your tip life, only flash when the machine asks to be flashed. Otherwise, it is difficult to know who flashed last. Over flashing will degrade tip performance and be the cause of tip replacement much more frequently than a blown tip due to a microdischarge. Flash Int. is set at 2. When you flash the emmision should read atleast 25, in the thirties indicates that the tip needed to be cleaned.
I hope is helps,
Sally Stowe wrote: } Hi all, } I am after some advice on the care of the FE tip and vacuum } on a Hitachi 4500. } } 1. How often should one bake out? The manual recommends baking out } when the vacuum deteriorates. After about 8 months operation ours is } better than it was to start with - IP1&2 off-scale, IP3 at } 7x10-7 Pa. On the other hand many people seem to recommend baking at } fairly short intervals "whether it needs it or not". We are inclined } to a non-interventionist approach but are getting a bit nervous...any } advice? } } 2. What should the flash current intensity be? Ours started at around } 15-20 (and we sometimes flashed twice to get a reading in the high } twenties) but has steadily crept up and is now in the high forties. } Is this good, bad or indifferent? Is it perhaps related to question } 1? If it gets too high does it wreck the tip? We are generally } flashing once or twice a day. } } } cheers } Sally } ---------------------------------------------------------------------- } Sally Stowe |Email: stowe-at-rsbs.anu.edu.au } Facility Coordinator |Post: } ANU Electron Microscopy Unit |ANUEMU (RSBS) } Ph 61 6 249 2743 |Australian National Univ. } FAX 61 6 249 4891 |Canberra, } http://online.anu.edu.au/EMU/home.htm } |AUSTRALIA 0200
William R. McManus Electron Microscopy Facility Department of Biology Utah State University Logan UT 84322-5305
The nice thing about Vaytek is, they will configure the hardware (step motor and filter wheels to most microscopes whether it is upright or inverted. And they have software for either IBM or MAC. to capture the stacks of images and do 3D reconstruction. I think the bottom line is: You need a good scope and a good camera to make the jump from 2D to 3D worthwhile.
Bob Morphology Core
On Wed, 18 Jun 1997, Philip Oshel wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Listers, } } I just got this request from a non-microscopist collegue, and I know there } are people here better qualified to help him than I am. This would be for } light microscopy. I have sent him some information and URLs, but more } detailed ideas from experts would be appreciated. He is in Tasmania, so } Australian product sources would be particularly useful. } } } Do you know anything about using digital image "slices" to reconstruct 3-D } } (kinda) images? There was an article in TREE last issue, and I had some } } info on software from a company called Vaytek, but I'd like to know what is } } needed to set up the microscope. } } Alastair Richardson } } alastair.richardson-at-zoo.utas.edu.au } } Thanks. } } Phil } } } Sic Hoc Legere Scis Nimium Eruditionis Habes { } Philip Oshel } Station A } PO Box 5037 } Champaign, IL 61825-5037 } (217) 355-1143 } oshel-at-ux1.cso.uiuc.edu } *** looking for a job again ****************** } } } }
} I am after some advice on the care of the FE tip and vacuum } on a Hitachi 4500. } } 1. How often should one bake out? The manual recommends baking out } when the vacuum deteriorates. After about 8 months operation ours is } better than it was to start with - IP1&2 off-scale, IP3 at } 7x10-7 Pa. On the other hand many people seem to recommend baking at } fairly short intervals "whether it needs it or not". We are inclined } to a non-interventionist approach but are getting a bit nervous...any } advice?
We have a Hitachi S-800 which is basically the slightly older, analog version of your instrument. I bake out when the vacuum deteriorates, like IP3 at worse than 3x10-6, or when we get a lot of tip noise. We have multiple users and strange samples, so this happens about 3 times a year. It sounds like your vac is great, so I would *not* bake out! We occasionally have post-bakeout problems...
} 2. What should the flash current intensity be? Ours started at around } 15-20 (and we sometimes flashed twice to get a reading in the high } twenties) but has steadily crept up and is now in the high forties. } Is this good, bad or indifferent? Is it perhaps related to question } 1? If it gets too high does it wreck the tip? We are generally } flashing once or twice a day.
CHeck with your Hitachi field service person. I don't think the intensity should be creeping up. I had this problem once, plus some other stability problems, which were solved with the replacement of the high voltage cable. The intensity is adjustable via a small pot on one of those boards under the 'scope - call Hitachi for advice. Generally you should see the need to flash about every 4 hours of use. A good indication of the condition of your tip is the extraction voltage, V1. Remember, the closer you get to 6.3KV, the blunter your tip (if your 'scope is like mine). There is a relationship between flashing intensity, V1, resolution, and tip condition. E-mail me if you need more details.
Do you love your FESEM like I love mine?!
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
I have a TEM tech position open. The job includes negative staining, thin sectioning, maintenance of laboratory (solutions, ordering, etc.), recording and filing related to College of American Pathologists certification, working with clinical samples looking for viruses, working with research samples looking at or for almost anything. Salary: $10.23/hr. Serious inquiries may be sent directly to me via email (saram-at-ac.pub.duke.edu) not to the server, or call me--see below.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Microscopy & Microanalysis '97, the joint Annual Meeting of the Microscopy Society of America, Microbeam Analysis Society and The Histochemical Society, will be held August 10-14, 1997, at the Convention Center in Cleveland, Ohio.
Please note the following deadlines:
July 7 Hotel Reservations July 15 Advance Registration at Reduced Rate
8,100 Meeting Information Pamphlets were mailed to members of the microscopy and microanalysis community in the United States on June 1. The Pamphlet contains a meeting-week-at-a-glance calendar, descriptions of special events and social events, and an Advance Registration Form.
If you have not received a Meeting Information Pamphlet but would like to, contact Microscopy & Microanalysis '97, particulars listed below. Please provide your fax number for quickest response. Or visit the Meeting web site at www.bright.net/~strecker/msno/mm97.html
Limited commercial exhibit space remains. Companies and organizations considering an exhibit should contact Microscopy & Microanalysis '97 as soon as possible. -- ********************************************************** * Microscopy Society of America * * 4 Barlows Landing Rd., Suite 8 * * Pocasset, MA 02559 * * Toll Free: 800-538-3672 * * Phone: 508-563-1155 * * Fax: 508-563-1211 * * Email: BusinessOffice-at-MSA.Microscopy.Com * * URL: http://WWW.MSA.Microscopy.Com * **********************************************************
Dear list members, In the next couple of weeks I have to make brass (70% Cu, 30% Zn) TEM specimens. Some of the samples will come from heavily deformed brass. Even though I have very limited reference books and journals available to me, I thought that it would be trivial to find electropolishing solutions and conditions for brass. Surprisingly I was wrong.
I am still looking up articles, but I would appreciate it if anyone can e-mail me recipes for electropolishing solutions and/or conditions they have used for this material. Maybe this material is listed in some reference books you have?
Please e-mail me privately. I am trying to get on the list, but have not been successful yet. I will post a summary of the replies to the list (if there is interest).
Position Available: The Analytical Imaging Facility of the Albert Einstein College of Medicine has an opening for an experienced electron microscopy technician. The laboratory is a comprehensive microscopy facility offering technologies that include transmission and scanning electron microscopy, fluorescence microscopy, histology, digital light microscopy and confocal microscopy.
Qualifications: The successful applicant must be well versed in a wide variety of microscopic methods, with at least two years of electron microscopy experience. BS degree minimum, MS degree and MSA certification desirable. The preferred candidate must be proficient in sample preparation techniques for transmission and scanning electron microscopy including: embedding, ultrathin sectioning, critical point drying, vacuum evaporation and photographic and digital image archiving. Operating knowledge of transmission and scanning electron microscopes as well as light microscopes for brightfield, phase contrast and fluorescence imaging is essential. Experience in histology, cryo EM, immunogold EM, video and digital imaging, confocal microscopy and image analysis software desirable. The applicant must have good communicative skills and the ability to work well with many people in a multi-user environment.
Duties: All aspects of specimen preparation of a variety of biological samples for TEM, SEM, and histology. All aspects of photographic and digital image archiving and analysis. Operation and routine maintenance of transmission and scanning electron microscopes, a variety of light microscope imaging stations and related laboratory equipment. Instruction of new users on all facility equipment, ordering supplies, and record keeping.
The position is full time with a full University benefits package and is available immediately. Applications will be accepted until the position is filled.
Please send CV, salary requirements and names of three references to Frank Macaluso, Analytical Imaging Facility, Albert Einstein College of Medicine, 1300 Morris Park Avenue, Bronx, NY 10461. fax: (718) 430-8996; e mail: macaluso-at-aecom.yu.edu **************************************************************************** Frank Macaluso tel: 718-430-3547 Analytical Imaging Facility fax: 718-430-8996 Albert Einstein College of Medicine e-mail: macaluso-at-aecom.yu.edu 1300 Morris Park Avenue Bronx, NY 10461 ****************************************************************************
As a side project I would like to attempt to do some immunocytochemistry and I would like to know if there is commercially available any DNA binding proteins directly conjugated to colloidal gold. In my former job I was a microbiologist at the NIH and I did PAg labelling on thin sections, so I am familar with the technique. These last six years I have made a change to materials science so I am a bit out of touch with what is commerically availble as far as gold probes are concerned. Another question I have - I already have nicely prepared blocks of the samples that I want to label but the resin I used was Durcopan (I could not dehydrate my samples with acetone or propylene oxide, that is why I chose the Durcopan resin). Do I have to start over again and use something like the LR White (that is what I used when I was at NIH) or is there some sort of etching that I can do to my sections in the Durcopan so that I can just label them? Thank you all in advance. This newsgroup is great!
Cheers, Peggy
Margaret E. Bisher
NEC Research Institute 4 Independence Way Princeton, NJ 08540. Tel.: (609) 951-2629 Fax: (609) 951-2496 e-mail: peggy-at-research.nj.nec.com
PS: Any commercial vendors should reply directly to me as per Nestor's request in the bylaws of this Newsgroup.
Does anyone have the number of the enlarger manufacturer called Simmon Omega (Or whatever it is called now). I have an old dichroic enlarger that I am trying to get information on.
Thanks in advance,
Michael Coviello The University of Texas-at-Arlington
Noesis Vision will be displaying such a 3d reconstruction package at the Microscopy show in Cleveland in August.
Literature will be available within a couple of weeks.
At 08:32 AM 6/18/97 -0500, Philip Oshel wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
---------------------------------------------------------------------------- ------------- Luc Nocente Tel: 514 345 1400 Noesis Vision Inc. Fax: 514 345 1575 e-mail: ln-at-noesisvision.com http://www.noesisvision.com 6800 Cote de Liesse, Suite 200 St-Laurent, PQ H4T 2A7,Canada ---------------------------------------------------------------------------- -------------
We are interested in finding (purchasing) a used SEM which is 10 or less years old to replace a very old SEM in the Biology Dept. Microscope Facility of Univ. of Calif, San Diego. Price is also a factor. We will also need know maintenance and performance history. Please respond to me at the addresses and numbers listed below.
Sincerely, Gina Sosinsky
Director, Biology EM Facility
***************************************** * Gina Sosinsky * * Department of Biology 0322 * * University of California at San Diego * * 9500 Gilman Drive * * La Jolla, CA 92093-0322 * * 619-534-6264 (phone) * * 619-534-0053 (fax) * * gsosinsky-at-ucsd.edu (email) * *****************************************
Philip Oshel wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } I should add, please reply directly to Alastair at his email address given. } } Listers, } } } } I just got this request from a non-microscopist collegue, and I know there } } are people here better qualified to help him than I am. This would be for } } light microscopy. I have sent him some information and URLs, but more } } detailed ideas from experts would be appreciated. He is in Tasmania, so } } Australian product sources would be particularly useful. } } } } } Do you know anything about using digital image "slices" to reconstruct 3-D } } } (kinda) images? There was an article in TREE last issue, and I had some } } } info on software from a company called Vaytek, but I'd like to know what is } } } needed to set up the microscope. } } } Alastair Richardson } } } alastair.richardson-at-zoo.utas.edu.au } ^^^^^^^^^^^^^^^^^^^^^^^^^^ } } } } Thanks. } } } } Phil } } } Sic Hoc Legere Scis Nimium Eruditionis Habes { } Philip Oshel } Station A } PO Box 5037 } Champaign, IL 61825-5037 } (217) 355-1143 } oshel-at-ux1.cso.uiuc.edu } *** looking for a job again ******************Alistair, There is another way. Differential Interference Contrast produces very thin optical sections which have been used very successfully with packages which construct 3-D images from serial sections. If you have a steady hand, you can move the stage (upwards is best) in specific increments. See what markings you have on your fine focus. Usually, it is marked in 0.2um increments (some are as fine as 0.1). These should be fine for most samples and may even be overkill for more gross structures, especially when viewed at lower magnifications.
Alternatively, you can have a Z drive installed on your fine focus and, under software control (ex: some of the image analysis packages like Media Cybernetics' Image Pro Plus have this sort of acquire-move the stage-acquire capability), again, move the stage upwards by specific increments and collect the stack of images.
Both of these alternatives are much less expensive than investing in confocal. The latter approach is better suited to those applications where the objects present a great deal of haze and glare and you really need the extra optical boost to image the true structures.
Good luck.... and let me know how things turn out.
Barbara Foster
The views expressed here are entirely my own and not intended to convey any commercial bias.
We are interested in finding (purchasing) a used SEM which is 10 or less years old to replace a very old SEM in the Biology Dept. Microscope Facility of Univ. of Calif, San Diego. Price is also a factor. We will also need know maintenance and performance history. Please respond to me at the addresses and numbers listed below.
Sincerely, Gina Sosinsky
Director, Biology EM Facility
***************************************** * Gina Sosinsky * * Department of Biology 0322 * * University of California at San Diego * * 9500 Gilman Drive * * La Jolla, CA 92093-0322 * * 619-534-6264 (phone) * * 619-534-0053 (fax) * * gsosinsky-at-ucsd.edu (email) * *****************************************
The 2-day symposium " {bigger} Materials Applications of Electron Holography and Related Techniques" (Session HH) {fontfamily} {param} Helvetica {/param} will be held at the Fall MRS meeting, Dec. 1-5, 1997. There are still a few days to prepare and submit abstracts. Please check the MRS website at http://www.mrs.org for further details, and use the website for submission of abstracts. The *absolute deadline* for abstract submission is June 23 (next Monday)...absolutely *no* abstracts will be accepted by MRS after this time.
There is an outstanding list of invited speakers for this symposium, and a number of excellent contributed papers have already been received. A few additional contributed papers will be considered, so please take this opportunity to submit your abstract. The MRS website provides a simple mechanism for abstract submission, and it only takes a few minutes.
Funds may be available to help support students/post-docs who are principal authors. Please submit requests for support to any organizer.
I have been following with interest the discussion on file format translation over the last few days. ANS is new to the EDS field. We will be introducing a line of systems at M&M '97.
Our software incorporates a file translator which can import several formats and save them in simple ASCII or our binary format. We are in the process of adding MSA ASCII format to both import and export sides.
We are prepared to add ANY widely used format to the translator DLL and make the DLL and a small translator application available on our web site FREE to anyone who would like it.
If you have a file format you would like to see added please send complete documentation to me and some sample spectra I can use for testing. You can e-mail the information to me at: bhardy-at-qtmsys.com
If you need to use anonymous ftp instead please use: ftp.qtmsys.com/pub/incoming/ and send me an e-mail to let me know it's there.
or mail it to me at:
American Nuclear Systems, Inc. 12633 Red Canyon Road Knoxville, TN 37922 423-671-0292 FAX 423-671-0293
Mike Coviello wrote =============================================== Does anyone have the number of the enlarger manufacturer called Simmon Omega (Or whatever it is called now). I have an old dichroic enlarger that I am trying to get information on. ================================================ I believe the firm you are looking for is the following:
Like in the world of microscopy, consolidations and corporate take-overs are happening in the photographic industry as well. But I believe these are the people you are looking for.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Jean - Pioloform and formvar require carbon evaporation on the film to make it stable under the electron beam. Butvar is pretty good without. Carbon only films a more trouble to make or more expensive to purchase. If the UA is too thick that too will cause trouble because a lot of heat is generated in such electron dense material. Apply and blot about six times by touching the grid against a drop of 'stain' and finish by blotting, well but not blotting off all of the solution. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 350+ Links, MSDS ************************ http://www.proscitech.com.au
} Hi there! } } I'm using uranyl acetate to get a negative stain of my material, that on } pioloform (0.5%) and I've even tried Formvar (0.5%) coated grids. } } Now my problem is this: the stain heats-up very fast, and blows my } coating, too often before I get a chance to take a picture. } } Should I try a more concentrated pioloform solution (because I found } Formvar to be even less stable) or does anyone know of a tougher coating? } } (I work at 75-100kV, most of the time at 50,000 X) } } Thanks for any advice! } } ???????????????????????????????????????????????????????????????????????????? ? } } "Life is the leading cause of Death" -B.C. } } Jean Le Clerc } } Institut de Recherche en Biologie Vegetale } } leclercj-at-magellan.umontreal.ca } Voice: 514-277-7938 } FAX: 514-277-7938 *call first* }
After dehydration did you Critical point dry (CPD) your cells? If you did not then that's your problem.
Looking at your protocol, I would like to suggest the following :-
1. Fix Marine phytoplankton cells in 2,5% glutaraldehyde in 0,1M sodium cacodylate buffer pH 7.2 for 1hour 2. Wash in 0.1M sodium cacodylate buffer pH 7.2 for 2 X 5 minutes 3. Post fixation - 2% osmium in 0.1M sodium cacodylate buffer for 1 hour 4. Wash as in (2) : Before dehydration transfer pelleted cells into a specimen processing capsule. They are 13mm diameter and 18mm high and have very small holes in each end, permitting liquid exchange, but which tend to retain a small amount of solution thereby reducing surface tension effects. Specimen can therefore be retained in the "wet" state up to dehydration and the CPD process. The capsule are very useful for CPD small specimens like Marine cells etc. (available from Agar - G 3314 catalogue no.). Place the closed capsules into no. 2 pill vials and change ethanol solutions in the pill vials using pasteur pipettes.
5. Dehydration: Ethanol 10% 30% 50% 70% 90% - 2 x 5 minutes each 6. 100% 3 x 5 minutes.
7. The capsules containing cells are than transferred into the CPD chamber and critical point dried with liquid carbon dioxide. After CPD mount specimen, coat and view.
Please note: do not wash with distilled water. Your salts will wash off during fixation and buffer rinses
all the best.
Vijay H Bandu Centre for Electron Microscopy University of Natal Private Bag X01 Scottsville 3209 South Africa
e- mail bandu-at-emu.unp.ac.za telephone : 0331 2605157 fax 0331 2605776
Try exporting to EMSA interchange format. That's available and I use it myself for this purpose.
Peter Statham
In article {"0015012D."-at-ccmail.pnl.gov} , John S Vetrano {js_vetrano-at-ccmail.pnl.gov} writes } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We are interested in the polytypism of SiC. Some references said that there are more than 200 polytypes in SiC. But we can only find details (such as 6H, 3R...) about 150 kinds. Is there anyone who knows the more details or where to find the details?
Any help would be greatly appreciated. Thanks.
Yaqiao Wu STATE KEY LABORATORY FOR FATIGUE AND FRACTURE OF MATERIALS INSTITUTE OF METAL RESEARCH CHINESE ACADEMY OF SCIENCES
At 11.14 17/06/97 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have a very good experience with Kodak Megaplus 1.6 (1500x1024 pixels approx.) Slow Scan CCD Camera fitted on a Philips CM120 TEM -35 mm camera port.
The camera is sw controlled by a PC + Windows. The framegrabber is a F-64 Board from Matrox, capable to grab images up to 1600x1200 pixels, but should exist also a cheaper solution from GrabBit.
The nice side of this solution is: you may use that camera also for an optical microscope !
I have no idea if exist also a version for Mac 8500. You may contact the following address:
SIS Soft-Imaging Software Corp. 2102 Beech Court Golden, CO 80401
Are you carbon coating your plastic? This makes an enormous difference because the plastic coatings are poor conductors of heat and electrons.
Other suggestions: How dry are the solvents that you use for making up your plastic solutions? If they are old bench reagents use fresh ones from a good supplier. Can you produce a relatively light preparation of negative stain? If its too thick have you tried a spreading agent such as bacitracin. Is the sample heavily contaminated with culture medium or excess extracellular material and can you wash the specimen to improve it? Have you tried a smaller mesh grid eg 400 mesh (400 linear squares to the inch or higher)?
You didn't say what you were looking at but for most preparations we get a way with carbon coated formvar (made from 0.3% formvar in chloroform) on 400 mesh copper grids
Good luck Malcolm Haswell University of Sunderland UK ----------
Hi there!
I'm using uranyl acetate to get a negative stain of my material, that on pioloform (0.5%) and I've even tried Formvar (0.5%) coated grids.
Now my problem is this: the stain heats-up very fast, and blows my coating, too often before I get a chance to take a picture.
Should I try a more concentrated pioloform solution (because I found Formvar to be even less stable) or does anyone know of a tougher coating?
(I work at 75-100kV, most of the time at 50,000 X)
Message-Id: {1.5.4.32.19970619130450.006bd010-at-biotech.ufl.edu} X-Sender: gwe-at-biotech.ufl.edu X-Mailer: Windows Eudora Light Version 1.5.4 (32) Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
We generally use pure carbon films that have ben floated off mica, with or without an underlying Formvar film.
Or you can carbon coat your Formvar, but a pure carbon film alone gives ther best reolution. } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } . At 05:31 PM 6/18/97 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
I have a bunch of Hitachi SEM filaments (tungsten) that need to be re-tipped. I checked Pella and EMS catalogs and they seem to offer re-tipping of everything BUT Hitachi filaments. Anyone know of a place that can help me out?
Gary Radice, Associate Professor gradice-at-richmond.edu Department of Biology 804-289-8107 (voice) University of Richmond VA 23173 804-289-8233 (FAX)
} I'm using uranyl acetate to get a negative stain of my material, that on } pioloform (0.5%) and I've even tried Formvar (0.5%) coated grids. } } Now my problem is this: the stain heats-up very fast, and blows my } coating, too often before I get a chance to take a picture. } } Should I try a more concentrated pioloform solution (because I found } Formvar to be even less stable) or does anyone know of a tougher coating?
Hi Jean,
I use 0.3% of pioloform for all my negative staining, I never have any problem like you described. I am wondering that you may make your supporting membrane too thin , such as you pull up the slide too fast from the solution. You also can check the tickness of the film on the basis of interference colors. Other than that, I may suggest you to use 300-400 mesh for negative staining.
Wish you luck,
*********************************************** * Ming H. Chen, PhD * * Medicine/Dentistry Electron Microscopy Unit * * University Of Alberta. * * Edmonton, Alberta, Canada * * * * Visit My Page At: * * http://www.ualberta.ca/~mingchen * ***********************************************
Our laboratory has had frustrating problems with paper jams in our Mitsubishi color video printer, model CP700U. Has anyone else had this problem, and do you have any solutions to offer??
The printer was purchased to accompany a new Philips XL-30 SEM with EDAX system. It only worked for about 2 months before the first paper jam. Once the paper jams, it cannot be re-threaded by the user to get it functioning again. The only option is to send the printer in to the repair center in CA, which takes about 2 weeks each time! This has happened a number of times over the past 8 months.
As if this wasn't bad enough, the latest repair produced a printer which, when powered on, causes disruptions to the SEM monitor image! Philips, as always, has been very helpful but the problem is not with the SEM, it is with the printer. Unfortunately, the Mitsubishi warranty/service department has been less than helpful, since their warranty policy only covers repairs, not returns. So we are stuck with trying to find a fix for it ourselves, since sending it in for repair has only made things worse.
This sort of situation is extremely frustrating. We would really like to get an idea of how often this happens with these roll printers. Does this happen with other models besides Mitsubishi? At this point we would love to just throw the thing in the trash and start over but we don't know if there is a better option out there. [The Sony SOUP-1200 is not compatible with our system].
Your comments or tips are appreciated, Karen
-- Karen Zaruba kszaruba-at-mmm.com 3M Company, 3M Center Bldg. 270-1S-01 (612)737-2971 St. Paul, MN 55144 fax: 736-1519 "The opinions stated above are my own, not necessarily 3M's"
Bill Tivol wrote: =================================================== Since brehmsstrahlung production goes up strongly with Z, higher production from these grids can only be due to the greater amount of material in the path of the beam. If the total mass of the grid can be reduced to the same level as that for other grid materials, brehmsstrahlung would be quite low. ==================================================== The carbon composite grids need their basic "thickness" in order to make them less fragile since they are made from an extremely brittle carbon material. A better alternative might be the diamond grids (actually made from diamond) which have a brehmsstrahlung level comparable to that of Be (even though the diamond grids are thicker). However these diamonds might not be your best friend since they are more expensive even than the Be grids . But the good news is that there are no toxicity questions with diamond (we don't believe there are problems with Be grids being used in a TEM either but that view is not shared universally).
Disclaimer: SPI offers both Be, diamond, and carbon composite grids and we really don't care which ones are used!
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Someone recently posted a question requesting information on Omega enlargers (sorry, I accidentally deleted the message). If this was a repair and/or parts related question, I have come across the following address for a business that specializes in repairing and rebuilding these. (Disclaimer---I have no financial interest in this company whatsoever):
Also, since most, if not all, EM labs have darkroom facilities and other photographic equipment, the following e-mail address may be useful:
http://www.fargo-ent.com
This site has an incredible wealth of information on all aspects of photo equipment repair, parts, and technical matters, as well as links relating to repair of almost anything imaginable.
Hope this is useful.
Randy Tindall Center for Electron Microscopy Southern Illinois University at Carbondale Carbondale, IL 62901
Gary Radice wrote: ================================================== I have a bunch of Hitachi SEM filaments (tungsten) that need to be re-tipped . I checked Pella and EMS catalogs and they seem to offer re-tipping of everything BUT Hitachi filaments. Anyone know of a place that can help me out? ================================================== We have offered retipping services for Hitachi filaments for some years and information about that service and current prices can be found on our website shown below. Turn around time is about 3 weeks after receipt of bases.
My guess is that the service might be offered by the other mentioned firms as well but for some reason the listing might have not made it into their catalogs.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
It is almost meeting time. As most of you know one of the big drawing cards at the FORUM booth in the convention hall is the availability of printed "Hints and Tips" for the microscopist. We need your help. Please support the TF by bringing with you some piece of methodology, a protocol, a discussion of a small convenience which you use in your labroatory in the form of 50 printed copies. It can be as short as a paragraph, or, if you like, several pages long. Please list your name and institution, e-mail, FAX, on the paper. Return the help which you have gotten from your colleagues over the years. Please bring your contribution to the TF booth in the convention hall on Monday or talk to me (I would love to talk to you - maybe we can go for a cookie, or cherry pie, or desert, etc., too) before the meeting. I will be arriving Sat noon before the meeting, and I will be staying at the Sheraton. Please, please, write some little thing. People who come by the booth are so appreciative of it. And - it is good for the reputation of TF. See you soon, Hildy
We are looking for a method to label cytochrome C oxidase on fungal hyphae in thin sectioned material. If anyone has experience with a technique to acconplish this, or knows of any relavent references, we would be very apprecative if you could pass along the information.
TIA
William R. McManus Electron Microscopy Facility Department of Biology Utah State University Logan UT 84322-5305
Gary Radice wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } I have a bunch of Hitachi SEM filaments (tungsten) that need to be } re-tipped. I checked Pella and EMS catalogs and they seem to offer } re-tipping of everything BUT Hitachi filaments. Anyone know of a place that } can help me out? } } Gary Radice, Associate Professor gradice-at-richmond.edu } Department of Biology 804-289-8107 (voice) } University of Richmond VA 23173 804-289-8233 (FAX)
Gary, We at Ladd would be happy to re-tip your Hitachi filaments. Our catalog # is 9-63025R for and the price is $12.95 each. Please contact me via e-mail or phone 1-800-451-3406 and I will tell you where to send them.
We are looking for a method to label cytochrome C oxidase on fungal hyphae in thin sectioned material. If anyone has experience with a technique to acconplish this, or knows of any relavent references, we would be very apprecative if you could pass along the information.
TIA
William R. McManus Electron Microscopy Facility Department of Biology Utah State University Logan UT 84322-5305
} } Hi all, } } We are looking for a method to label cytochrome C oxidase on fungal hyphae } in thin sectioned material. If anyone has experience with a technique to } acconplish this, or knows of any relavent references, we would be very } apprecative if you could pass along the information. } } TIA } } William R. McManus } Electron Microscopy Facility } Department of Biology } Utah State University } Logan UT 84322-5305 } } } In our laboratory we have devised a technique to detect chromophores or any compounds with an absorption in the visible region of the spectrum using electron energy loss spectroscopy (EELS).
How much of cytochrome C is contained in fungal hyphae? The compound could be localized directly (without a label) as long as it is present in a large enough concentration.
Also, what function does cytochrome C serve and what is the appeal in localizing this protein? (please forgive my ignorance on the subject)
Melanie
________________________________ Melanie Barfels Department of Medical Biophysics University of Toronto (416) 946-2000 XT 5185
Dear all Can you help me? I have been asked to look at some wax embedded tissue sections in the SEM and to do EDX analysis to detect gold in the sample. My problem is I no longer have access to a carbon coating unit ( mine recently died) and I am only able to sputter coat with gold - not a good idea if I want to detect gold in the sample!
Does anyone have any hints and tips as to how best to set up the SEM (a J6400) for low voltage so I can examine the specimen uncoated, prevent charging and analyse for Au? Another possibility is to examine the sample on a FEG-ESEM ( we are having one installed within the next few weeks). Any suggestions about this would be of help, but for quick results I have to use a conventional SEM. I also thought about TEM/EDX. Is it possible to re-embed wax sections for TEM and does anyone have a protocol for this?
All ideas will be much appreciated. Thanks very much
Nikki Bock ********************************************************************** Nikki Bock EM Technician Dept. of Materials Engineering & Materials Design University of Nottingham Nottingham NG7 2RD Tel: (0115) 9513759 Fax: (0115) 9513741 Email: emznjb-at-emn1.nott.ac.uk
Does anyone know of a stain, either for light microscopy or SEM, that could differentiate a gelled starch binder from a cellulose fiber matrix? If not a stain, any other good methods? The magnification would be 50X-300X, and the sample is dry.
Thanks --
Dave Stadden DRStadden-at-Armstrong.com 717-396-5109
Does anyone have a chrome coater in the neighbourhood???? 15 angstroms of chrome (Edwards Xenosput) won't get in the way of detecting gold.
} Can you help me? I have been asked to look at some wax embedded } tissue sections in the SEM and to do EDX analysis to detect gold in } the sample. My problem is I no longer have access to a carbon coating } unit ( mine recently died) and I am only able to sputter coat with } gold - not a good idea if I want to detect gold in the sample! } } Does anyone have any hints and tips as to how best to set up the SEM } (a J6400) for low voltage so I can examine the specimen uncoated, } prevent charging and analyse for Au? } Another possibility is to examine the sample on a FEG-ESEM ( we are } having one installed within the next few weeks). Any suggestions } about this would be of help, but for quick results I have to use a } conventional SEM. } I also thought about TEM/EDX. Is it possible to re-embed wax sections } for TEM and does anyone have a protocol for this? } } All ideas will be much appreciated. } Thanks very much } } Nikki Bock } ********************************************************************** } Nikki Bock } EM Technician } Dept. of Materials Engineering & Materials Design } University of Nottingham } Nottingham NG7 2RD } Tel: (0115) 9513759 } Fax: (0115) 9513741 } Email: emznjb-at-emn1.nott.ac.uk
Cheers :) George Department of Earth and Atmospheric Sciences University of Alberta Edmonton, Alberta T6G 2E3 Canada ph: 403-492-5746 fax: 403-492-2030
We were wondering if anyone out there is planning to upgrade their camera system and has an existing CCD camera they would like to sell. We are looking for one to attach to our JEOL 100CX. I would appreciate any information. Please send directly to my e-mail address. Thank you in advance. Cathy Kelloes
Karen, I have a Mitsubishi dye-sub printer that I have been happy with for a few years, but now it needs repair, and I agree with you in your opposition to mailing equipment across the country. Does anyone know of someone who repairs Mitsubishi printers in the Chicago area? Do I realy have to have to pack it and send it all the way to California? The next equipment I buy will have service in this area or no sale. Joyce Craig Chicago State University
Does anyone know of a source of small glass strips, about 3-5mm x 10-12mm, having the thickness of microscope slides (optimally) or coverslips?
What I need them for is to help in handling/weighting down adhesive tapes during chemical processing for biological TEM. I've found the tapes separate from ACLAR, double-stick tape and Permanox dishes during dehydration and infiltration, but they remain adherent to glass coverslips. However, most glass coverslips are too large to comfortably fit into my processing vials. I can always get our machine shop to cut up glass slides for me, but a commercial source might be more convenient.
Thanks as always for your help -- this list is wonderful! Karen
-- Karen Zaruba kszaruba-at-mmm.com 3M Company, 3M Center Bldg. 270-1S-01 St. Paul, MN 55144 "The opinions stated above are my own, not necessarily 3M's"
On 20 Jun 1997 David_R_Stadden-at-armstrong.com wrote:
} } Does anyone know of a stain, either for light microscopy or SEM, that could } differentiate a gelled starch binder from a cellulose fiber matrix?
Try Schiff's reagent (I get mine from Fisher, but I am sure many places can supply it). Then, do a control where you digest with alpha-amylyase followed by reaction with Schiff's. Any place on the specimen that was starch or glycogen would be magenta with just Schiff's and colorless (or faint pink) following digestion.
Please contact me directly for a full protocol.
Good luck.
Don
______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 The College of New Jersey fax: (609) 771-2674 Trenton, NJ 08650-4700
------------------------------------- To All Phillips Tansmission Microscope users
I have Phillips TEM and I am using LaB6 , the only problem is that in the past two years we seem to have used three filaments . I feel that this is a very short life for LaB6 "our vacuum is within specs,we have set up the filament spacing between the wehnelt and the tip, and we are not over-saturating" .What I would like to know is the other Phillips 420 users what type of LaB6 are you using and what sort of life do you get from your filament.
I see references for cleaning TEM/SEM metal parts such as Wehnelt caps by sonicating them in acetone or alcohol. Yet my sonicator manufacturer specifically warns against using these solvents in their sonicator. I understand why you wouldn't want to use water based solutions for cleaning these parts but I also don't want to blow up the lab by sonicating flammable liquids, if this really is cause for concern.
What do you people do?
Gary Radice, Associate Professor gradice-at-richmond.edu Department of Biology 804-289-8107 (voice) University of Richmond VA 23173 804-289-8233 (FAX)
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I put my dirty samples into a beaker, put the solvent in the beaker and then the beaker goes into the sonicator that has water in it. I don't use the solvent in the sonicator directly.
Margaret E. Bisher
NEC Research Institute 4 Independence Way Princeton, NJ 08540. Tel.: (609) 951-2629 Fax: (609) 951-2496 e-mail: peggy-at-research.nj.nec.com
Why not use a diamond scribe to score the slides or coverslips and break the? You can even use a straight edge to wind up with straight and square sides. Similar to old fashioned glass knives or as they do with window glass I have made small slivers from coverslips regularly this way.
Hope this helps, Louie
} Does anyone know of a source of small glass strips, about 3-5mm x } 10-12mm, having the thickness of microscope slides (optimally) or } coverslips? } } Karen Zaruba } kszaruba-at-mmm.com } 3M Company, 3M Center Bldg. 270-1S-01 } St. Paul, MN 55144 } "The opinions stated above are my own, not necessarily 3M's"
Louie Kerr Research and Education Support Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543 508-289-7273 508-540-6902 (FAX)
} Name: Anaspec } E-mail: anaspec-at-.icon.co.za } Date: 6/20/97 } Time: 5:38:56 PM } } ------------------------------------- } To All Phillips Tansmission Microscope users } } I have Phillips TEM and I am using LaB6 , the only problem is that in the } past } two years we seem to have used three filaments . I feel that this is a very } short life for LaB6 "our vacuum is within specs,we have set up the filament } spacing between the wehnelt and the tip, and we are not over-saturating" } .What I } would like to know is the other Phillips 420 users what type of LaB6 are you } using and what sort of life do you get from your filament.
I'd say it depends on how and for how long you are using your TEM. I would expect upto around 1,000 hrs from LaB6 used for fairly hard analytical TEM, so 3 filaments in 2 years sounds OK, unless you are doing low mag ( {20k), imaging when it is perhaps not so good.
Regards, Lary Stoter
-- Dr L. P. Stoter Technical Editor, MICROSCOPY & ANALYSIS Technesis M&A web site - 17, Rocks Park Road http://www.microrgc.demon.co.uk Uckfield, E. Sussex email: LPS-at-teknesis.demon.co.uk TN22 2AT Phone: +44 (0)1825 766911 United Kingdom Fax: +44 (0)1825 766911
Ultimately we would like to label F-actin at the EM level in growth plate chondrocytes and matrix vesicles. We have done some pilot experiments at the light microscope using:
1) Rabbit anti-actin antibody (A 2066 from Sigma) on paraffin sections. The label was confined only to a subset of smooth muscle cells in control tissues.
2) Phalloidin-BODIPY FL worked very nicely on cryostat sections, therefore we tried anti-BODIPY antibody, followed by biotinylated goat-anti-rabbit, Streptavidin peroxidase and AEC substrate. However, we got no labelling.
We would appreciate any input on F-actin labelling at EM level in non-muscle cells or any experience dealing with rabbit anti-BODIPY FL antibody.
Thank you,
Sarka Lhotak EM Facility, McMaster University Hamilton, Ontario, Canada
3 filaments within 2 years??? First answer to your question: Kimball, Denka, and FEI LaB6 filaments all have good track records in a variety of Philips scopes including the EM420. As for the short filament life, I have to present a couple of observations for your consideration. I am assuming both your HV2 vacuum is at least below 35 and that if the filament is kept saturated between specimen changes, the change in HV2 vacuum does not go so high as to open the V6 valve(HV2 reading approx. 80), AND that the HV2 recovery time to the 35 or below level is within spec. Also, the gun has been properly conditioned and occurences such as flash-overs, micro-discharges, or other gun instabilities are ok, then:
1) Is the LaB6 tip evenly wearing away in it's crucible? If so, then the mount of the tip and probably the whole filament itself is good. If this is the case, I would first suspect that the bias(Emission) current setting you are using is well above the typical 10-20 microamps which will severally shorten any filaments life. Some customers will sacrifice the LaB6 life to get more "light" or beam density, depending on the application, especially when using a smaller C1(Spot) probe or a C2(Condensor) aperture. If the emission reading appears within the typical 10-20 uA, have your service person monitor the actual current to the filament and compare it to the meter reading. If the feedback circuit is not working properly, your filament could be getting unknowingly overheated.
2) If the tip appears to have a good portion left, look under a light microscope to see if the crystal is loose or broken away from the crucible or lead-attaching holder. Also check the filament leads for unusual stress or fractures. This can indicate a poorly manufactured filament. It can also indicate problems occuring within the emission chamber.
Perhaps the easiest of all would be this suggestion: the above mentioned LaB6 manufacturers all give each filament a serial number. If you still have the 3 blown filaments and the manufacturer has labeled them in this way, send the bad ones to them for their assessment.
------------------------------------- To All Phillips Tansmission Microscope users
I have Phillips TEM and I am using LaB6 , the only problem is that in the past two years we seem to have used three filaments . I feel that this is a very short life for LaB6 "our vacuum is within specs,we have set up the filament spacing between the wehnelt and the tip, and we are not over-saturating" .What I
would like to know is the other Phillips 420 users what type of LaB6 are you using and what sort of life do you get from your filament.
3 filaments within 2 years??? First answer to your question: Kimball, Denka, and FEI LaB6 filaments all have good track records in a variety of Philips scopes including the EM420. As for the short filament life, I have to present a couple of observations for your consideration. I am assuming both your HV2 vacuum is at least below 35 and that if the filament is kept saturated between specimen changes, the change in HV2 vacuum does not go so high as to open the V6 valve(HV2 reading approx. 80), AND that the HV2 recovery time to the 35 or below level is within spec. Also, the gun has been properly conditioned and occurences such as flash-overs, micro-discharges, or other gun instabilities are ok, then:
1) Is the LaB6 tip evenly wearing away in it's crucible? If so, then the mount of the tip and probably the whole filament itself is good. If this is the case, I would first suspect that the bias(Emission) current setting you are using is well above the typical 10-20 microamps which will severally shorten any filaments life. Some customers will sacrifice the LaB6 life to get more "light" or beam density, depending on the application, especially when using a smaller C1(Spot) probe or a C2(Condensor) aperture. If the emission reading appears within the typical 10-20 uA, have your service person monitor the actual current to the filament and compare it to the meter reading. If the feedback circuit is not working properly, your filament could be getting unknowingly overheated.
2) If the tip appears to have a good portion left, look under a light microscope to see if the crystal is loose or broken away from the crucible or lead-attaching holder. Also check the filament leads for unusual stress or fractures. This can indicate a poorly manufactured filament. It can also indicate problems occuring within the emission chamber.
Perhaps the easiest of all would be this suggestion: the above mentioned LaB6 manufacturers all give each filament a serial number. If you still have the 3 blown filaments and the manufacturer has labeled them in this way, send the bad ones to them for their assessment.
------------------------------------- To All Phillips Tansmission Microscope users
I have Phillips TEM and I am using LaB6 , the only problem is that in the past two years we seem to have used three filaments . I feel that this is a very short life for LaB6 "our vacuum is within specs,we have set up the filament spacing between the wehnelt and the tip, and we are not over-saturating" .What I
would like to know is the other Phillips 420 users what type of LaB6 are you using and what sort of life do you get from your filament.
} To All Phillips Tansmission Microscope users } } I have Phillips TEM and I am using LaB6 , the only problem is that in the } past } two years we seem to have used three filaments . I feel that this is a very } short life for LaB6 "our vacuum is within specs,we have set up the filament } spacing between the wehnelt and the tip, and we are not over-saturating" } .What I } would like to know is the other Phillips 420 users what type of LaB6 are you } using and what sort of life do you get from your filament.
We have been using a Philips 400T with a LaB6 for over ten years. Typically we obtain lives of twelve to eighteen months. We found the key to obtaining this longevity was to leave the HT on all the time with at least sixty percent of the saturation current, this keeps the LaB6 crystal hot preventing thermal cycling where most of the damage to the crystal can occur. To prevent excessive wear on the viewing screen we leave the microscope at maximum magnification and intensity (C2) fully clockwise.
We try to limit emission current to less than 15 microamperes. Early on leaving the HT on all the time did cause some problems with oil leaking from the HT tank cable well, but our service engineer replaced the seal and that problem was solved. It is also necessary to turn off the HT for camera changes, but if the camera is changed within a reasonable time (under 10 minutes) the HT can be brought back up to 120 KV in under a minute while watching HV2, and returned to sixty percent saturation. Of course if the gun is brought to atmosphere for any reason, or allowed to cool due to utility shut downs then an extended time is required to bring the LaB6 back up to 120 KV and reheated. The only problems we have experienced with running the LaB6 this long is that; 1. usually the last few weeks you can experience instabilities. 2. We remove the anode and clean at each filament change. 3. Replace wehnelt aperture each filament change.
Gatan Duomill Tip; To save expense of replacing sample holders we use carbon double sticky tape made for SEM samples to protect the sample holder posts from milling, you still have to replace the plates but you don't have to keep buying the complete holder when the threads are milled away.
I think all sonicator manufacturers include a "do not use with flammable solvents" warning, and all EM labs probably sonicate something in acetone or alcohol. Of course the primary liquid in the sonicator is water and the flammable solvents are in a beaker. We use ours in a hood.
As far as water based solutions for cleaning - we've been using (for 10+ years) a "Mr. Clean"/water solution for removing metal polish from our electron gun cartridges (followed by water, methanol, acetone, methanol) with no problems.
At 02:54 PM 6/20/97 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have been asked to study electrodeposited copper microstructure. My goal is to determine the relationship among electrodeposition process operational variables (electrolyte concentration, impurities deposition, aditives, current intensity applied etc.), microstructure (texture, grain size...), and mechanical properties of the metal.
You are all welcome to visit 5 copper micrographs I have posted on the Internet and please fell free to make any comments on them:=20
http://metallography.com/marti.htm
Any ideas will be welcome. Any reference info on the specific subject of electrodeposited copper microstructure will also help me. Can you tell me where to find copper cathode micrographs?
Please reply to Juan Marti at: jmartip-at-cepade.es
The descriptuion of the pictures is as follow:
The 5 micrographs correspond to the same cathode sample and show different structures among which I am not able to determine which one is the real one. I hope that you may help me in the interpretation of these images:
- Bottom left picture (cobre4). Etched with HNO3 (25%) at 70=BAC during only 5 seconds. Longitudinal section of the cathode showing what seems to be the grain boundaries. Lens Objective: 50x.
- Upper left and right pictures (cobre1). Etched with HNO3 (25%) at 70=BAC during 15 seconds. Longitudinal section of the cathode. It apparently shows big irregular grains but I=92m not sure if the sample might be under etched, thus not showing the real microstructure. Lens Objective: 50x.
- Middle right picture (cobre3). Etched with HNO3 (25%) at 70=BAC. More etching time. Longitudinal section of the cathode showing much smaller grains. Grains also seem irregular. Lens Objective: 50x.
- Middle left picture (cobre2). Etched with HNO3 (25%) at 70=BAC. Transversa= l section of the cathode showing what appears to be large grains grown in the current flow direction. Lens Objective: 20x.
- Bottom right picture (cobre6). Shows detail of abnormal grain size. Lens Objective: 20x.
Among the first 3 micrographs (cobre4, cobre1 and cobre3) can you tell which one is the real pure copper microstructure?
Any ideas will be welcome. I would also appreciate any help on specific references to copper cathode microstructures descriptions and micrographs.= =20
Thanks in advance.
Please reply to Juan Marti at: jmartip-at-cepade.es
Micrographs web site at: http://metallography.com/marti.htm
Hi, It sounds like these creatures may be better observed without fixation in their own environment. Would the use of an environmental microscope be better than a low-vacuum system? In an environmental SEM or ESEM, 100% relative humidity can be maintained allowing observation of living creatures and water... The Philips/Electroscan ESEM is the only system that can do this.
Back in February I asked for names of labs where we could outsource some of our polymer work (cryo, staining, TEM). I was asked to compile such a list by our management. Quite a few professional labs and some Universities were kind enough to answer. I was in the process of retrieving and archiving this information when my computer crashed. I was able to save the names of the commercial labs but I lost all of the information on the universities. So, I would appreciate it if Universities that accept contract work (on a per sample basis) for TEM of polymers would send me their address, contact name and tel. no. Please send answers to the following e-mail address:
Sorry, I'm late in picking up this thread. Jan's comment about avoiding fixation sparked a thought. Perhaps you might inquire about the Leitz Elsam accoustic microscope. Specimens are kept in a fluid (possibly seawater?) and scanned by a sound wave focussed onto the specimen by a sapphire lens. The resultant accoustic map looks very similar to a electron micrograph.
I don't know that there are many around so you would need to locate one (the List would undoubtedly be a good place to start) and ask whether the obtainable magnifications would be appropriate for your work.
Just a thought,
Geoff Avern Manager Microscopy Labs Australian Museum Sydney, Australia
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi, It sounds like these creatures may be better observed without fixation in their own environment. Would the use of an environmental microscope be better than a low-vacuum system? In an environmental SEM or ESEM, 100% relative humidity can be maintained allowing observation of living creatures and water... The Philips/Electroscan ESEM is the only system that can do this.
There was a recent posting re: carpal syndrome among microscopists. At the time it was not of special interest to me and apparently I did not save it. Yesterday a friend asked me about the subject so it turns out to be of interest after all. If anyone can give me the reference or resend me (personally) a copy of the posting, I would appreciate it.
Dear all, Thanks very much for all the advice about my wax samples. I've now found someone locally who can carbon coat my samples until our coater is repaired, so my problems are solved. Thanks again. Nikki ************************* Nikki Bock Dept. ME&MD University of Notingham Nottingham NG7 2RD Email: EMZNJB-at-EMN1.NOTT.AC.UK
--------------------------------------------------------------------- -- The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear all, Thanks very much for all the advice about my wax samples. I've now found someone locally who can carbon coat my samples until our coater is repaired, so my problems are solved. Thanks again. Nikki ************************* Nikki Bock Dept. ME&MD University of Notingham Nottingham NG7 2RD Email: EMZNJB-at-EMN1.NOTT.AC.UK
Hi Folks, a post-doctoral position is available within the Center for biologic imaging at the University of Pittsburgh Medical School to study the biology of dystrophin and related proteins in skeletal muscle. This fellowship is directed to furthering our understanding of how dystrophin is involved in the development and maintenance of the muscle fiber in health and disease. The principal approaches to be taken in this project will be a combination of light (live cell, confocal, etc) and EM (immuno-EM and freeze-fracture) techniques. If you are interested or know someone who might be please forward applications and CVs by email to this address Thanks Simon
-- Simon C. Watkins Ph.D. Associate Professor Director CBI University of Pittsburgh Pittsburgh PA 15261 tel:412-648-3051 Fax:412-648-2004 URL:http://sbic6.sbic.pitt.edu
----------------------------------------------. } } } } } } Hello, } } } } } } I'd like to know how could I get histological specimens from } } } titanium implants inserted into rabbit bone. The problem is cutting } } } bone with metal. } } } It seems that the suitable equipment is named Exact. How does it } } } work? What about its price? Where can I get it? } } } Good morning, } the address for EXACT that I have is 5100 N Brookline } Suite 740 } Oklahoma } City, Ok. 73112 } phone: (405) 943-4441 } } I believe the system costs about $80,000.00. We have been cutting } titanium and glass implants in various models for 12 years using the Leitz } 1600 saw microtome. I understand they have a new model on the market that } is very fine. the price is much less than the EXACT system. Depending on } what you need to do I would look at both instruments. Leitz address is E. } Leitz } } Rockleigh, New Jersey } } phone:(207) 767-1100 } } hope this helps. Mary Stone } } } Yours sincerly, } } ^-^^-^^-^^-^^-^^-^^-^^-^^-^^-^^-^^-^^-^^-^^-^^-^^-^^-^^-^^-^^-^^-^ } Mary Stone ph: (904) 462-0861 } Research Histology Core fax: (904)462-0875 } 12085 Research Drive } Alachua, FL 32615 } email: marystone-at-mailman.biotech.ufl.edu } ^-^^-^^-^^-^^-^^-^^-^^-^^-^^-^^-^^-^^-^^-^^-^^-^^-^^-^^-^^-^^-^^-^ } } } } ******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
About a dozen of you responded to my query about what solvents to use when sonicating EM parts for cleaning. I thought I would summarize the responses.
Most of you indeed sonicate in acetone and alcohol (and one brave soul has sonicated ether!) but use these solvents in beakers suspended in or sitting on the bottom of the water-filled sonicator tank. Those who sonicate flammable solvents always do this in the fume hood, to avoid build-up of fumes that could be ignited by a spark from the sonicator electronics. Some people use cool water to reduce vaporization of the flammables but this also probably reduces the efficiency of sonication. Sonication times vary from a few minutes to a half hour or more. No respondent reported ever having a problem sonicating these solvents.
A couple of respondents use "Mr. Clean" as the initial sonication solvent, followed by water, acetone, then ethanol or methanol. (For those outside North America, Mr. Clean is a liquid, general purpose household cleaner often used for washing floors and walls).
One respondent highly recommended sonicating in 5-10% dilute ammonia, and also recommended avoiding metal polishes that offer "long life" because they leave an anti-oxidation residue that is difficult to remove.
One repondent uses a commercial sonicating solution for metals sold by Ladd.
Thanks to all who offered advice.
Gary Radice, Associate Professor gradice-at-richmond.edu Department of Biology 804-289-8107 (voice) University of Richmond VA 23173 804-289-8233 (FAX)
I am interested to obtain some information about WDX system with SEM or LVSEM. What's the quality of image at high probe current and what's the evolution of sample (damaged or not). Is it possible to decrease damage if I use a cryo system? Thanks in advance for your help (comments, references...).
Best regards to all Didier
-------------------------------------------- Didier Le Thiec I.N.R.A. Centre de Recherches Forestieres Unite d'Ecophysiologie Forestiere Laboratoire de Pollution Atmospherique 54280 Champenoux - France
Recently, Jordi Marti requested information from Universities that accept contract work (on a per sample basis) for TEM of polymers.
As the proprietor of a commercial analytical service laboratory, I wish to point out that Universities which provide such services would be in violation of NSF Important Notice 91. This document recognizes that private analytical and testing labs make significant contributions to the technical and research infrastructure of the United States. Private for-profit labs produce a significant amount of innovative research and new analytical technology. When production processes are stopped, e.g. due to unknown contamination, manufacturing plants rely on the rapid turnaround provided by private analytical labs to get up and running again. However, the existence of this infrastructure is threatened when government-supported non-profit universities step outside their proper research role and improperly provide analytical services in competition with commercial laboratories.
Don Chernoff
Advanced Surface Microscopy, Inc. E-Mail: asm-at-indy.net 6009 KNYGHTON RD. Voice: 317-251-1364 INDIANAPOLIS IN 46220 Toll free: 800-374-8557 (in USA) web: http://www.a1.com/asm Fax: 317-254-8690 (If you experience difficulty in accessing our website, note that the web address uses numeral "1" in "a1")
A high probe current always means a large spot size with a W filament driven system. A large spot size will always provide an inferior image quality i.e. limited to the low thousands or even worse in a standard SEM=
or LVSEM.
Combining the EDX with backscattered images adds information about conten= t in the form of atomic number contrast. Also the relationship between the=
BSE image and the EDX analysis is much better in terms of similar reactio= n volumes when compared with the so called SE image. In LVSEM all these points are enhanced by the lower charge situations.
Low charge will mean less damage but the "damage" will relate to the specimen type e.g. high probability - polymers, waxes, animal tissue: low=
(zero) probability - metals and ceramics.
There is no substitute for small working distances ( {10mm but great care with a BSE detector) when trying to obtain the highest specimen currents,=
in this case the lower aberration coefficients improve the spot size for you and the exclusion of external fields also help to reduce the probe si= ze improving the image quality.
A high probe current always means a large spot size with a W filament driven system. A large spot size will always provide an inferior image quality i.e. limited to the low thousands or even worse in a standard SEM=
or LVSEM.
Combining the EDX with backscattered images adds information about conten= t in the form of atomic number contrast. Also the relationship between the=
BSE image and the EDX analysis is much better in terms of similar reactio= n volumes when compared with the so called SE image. In LVSEM all these points are enhanced by the lower charge situations.
Low charge will mean less damage but the "damage" will relate to the specimen type e.g. high probability - polymers, waxes, animal tissue: low=
(zero) probability - metals and ceramics.
There is no substitute for small working distances ( {10mm but great care with a BSE detector) when trying to obtain the highest specimen currents,=
in this case the lower aberration coefficients improve the spot size for you and the exclusion of external fields also help to reduce the probe si= ze improving the image quality.
} Does anyone know of a source of small glass strips, about 3-5mm x } 10-12mm, having the thickness of microscope slides (optimally) or } coverslips? }
} Karen Zaruba } kszaruba-at-mmm.com } 3M Company, 3M Center Bldg. 270-1S-01 } St. Paul, MN 55144 } "The opinions stated above are my own, not necessarily 3M's"
Hi Karen,
They may be a bit large, but leighton tube cover slips are available from Fisher, Corning, etc. I have some old ones from Bellco Glass which are 9x22mm.
good luck ed
Edward J. Basgall, PhD The Pennsylvania State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618
I am doing pre-embedding EM-ICC using Vectastain ABC with DAB detection. The ICC was performed after the primary fixation, then the tissue (2-3 mm cube of golfish retina) was fixed in osmium with potassium ferricyanide. Everything looks very promising, except I would like to generate a DAB product with a little more contrast. We did not do an Nickle choride or any other types of amplification. Any suggestions?? Linda Barthel Research Associate II Department of Anatomy and Cell Biology University of Michigan lab (313) 764-7476 fax (313) 763-1166 barthel-at-umich.edu
Thanks to all those who answered my posting requesting information about outsourcing labs.
I also thank those of you who brought to my attention NSF 91, I was not aware of it [ although I should have been] and I fully agree with your position. It would be my expectation that those universities that do perform contract work would honor the requirements of NSF 91. At the same time I should point out that some of the responses came from universities outside of the US . Furthermore, the list of EM service providers published by "Microscopy and Analysis" contains a number of such universities.
I like to think that, in the majority of cases, the contract work assigned by corporations is determined mostly by factors such as the qualifications of the EM laboratory ,the quality of the work performed, the dependability of the service and ( these days) the turn-around -time. It is in some of these factors where the private EM labs can have an edge over university facilities since these [the universities] are quite often involved in long term research and therefore do not have the flexibility to accommodate the short term needs of corporations.
Hello, I have just subscribed to the group and am asking if there are any Amateur Microscopist on this list? From what I can see it is mainly professional especially when the discussion is on the electron variety! I am an amateur in the UK. If anyone knows of a list that is more for Amateurs I would be grateful too.
Also where is the best type of Environment to find an Amoeba! I can find all the other types of protozoa (such as Paramecium and coleps) yet have failed to come across this text book example!
Thanks in advance
Conrad Perfett
------------------------------------------------------------------------ ----------- "Any sufficiently advanced technology is indistinguishable from magic" ----------------------------------------------------------- Arthur C Clarke ----
I run an old Cameca MBX microprobe. I currently have a faulty bargraph display on SP1. I've troubleshot the board, and have found a faulty bargraph display chip. The component is a Burroughs "Self Scan Bar Graph', (BG12201-2).
As all I need to replace is the above component, I was hopeful somebody out there may know how I can contact Burroughs.
In one form or another commercial services are at most Universities, that's a fact. Various methods/terminology has been established to avoid "unfair competition". The often quoted NSF Important Notice 91 should be read by anyone in the interest of what it really means. First of all it only applies to NSF purchased equipment and it is only a" guideline". According to an article in "Science", June 83 "no penalities are mentioned for violations of the guidelines, and NSF is apparently counting on the effect of clearly serving notice on the issue to avoid trouble". Noteing the above , and the reality of what is going on, it seems useless to continue to play a cat and mouse game (ie--commercial companies threating with the NSF rule and Universities forming alternate paths to avoid the legal terms of unfair competition). Ultimately the customer will make the choice as whom to use based on the services available--and they are different!! Commercial companies should note that there is not a conspiracy out there by the EM operators to grab their business. In fact you have friends on the inside of Universities who have spent countless hours trying to keep uninformed administrators from trying to make a quick buck. These same people have also directed customers to you without your knowledge.Many Universities also seem to have good working relationships with industry through state approved programs to assist their citizens and build a strong economy. Service facilities probably could find some way to interface with these programs for everyone's benefit. On the other hand University personnel should be sensitive to unfair prcatices and stick within establised guidelines.Also keep in mind potential conflicts of interest.
The above statements are only my personal opinion as a person who has been on both sides of the fence. These statements have not been approved by the University of Maryland in any way shape or form. I have no intention of starting, partcipating in futher discussion, or responding to follow ups. I no longer read the MSA bulletin board so if you want to contact me please do so directly , but I may not get back soon due to a very busy summer.
One possible solution to your problem is the Osmium-Thiocarbohydrazide-Osmium (OTO) method. I don't have the reference at hand, I remember it was a technical note in J. Histochem. Cytochem.in the mid-80's. It is quite simple: after regular osmication, soak the block with thiocarbohydrazide then incubate again briefly with osmium. I had to adapt their protocol for my materia. In particular, I found it necessary to rinse twice my cells with dd water between the steps to avoid unsightly precipitate. I hope this helps. Good luck, Michel **************************************************** Michel Deschuyteneer, Ph.D. deschuyt-at-sbbio.be Scientist Electron Microscopy Laboratory
SmithKline Beecham Biologicals Rue de l'Institut, 89 B1330 Rixensart, BELGIUM Tel: +32-2-656 9290 Fax: +32-2-656 8164 **************************************************** Standard disclaimer: the opinions expressed in this communication are my own and do not necessarily reflect those of SmithKline Beecham. ****************************************************
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Conrad: Any murkey pond is a possibility. But my former boss at U.C. Berkeley found the giant amoeba that he used for years in his research on the glass wall of the tropical fish tank in his office. There's an excellent new book on protozoa. You'll find it listed in the Project MICRO bibliography on MSA's Web page, along with many others that will interest adult amateurs. Here's the description:
Anderson, R. and Druger, M., eds. 1997 Explore the World Using Protozoa 240pp, paperback, 8.5x11", $29.95. ISBN 0-87355-159-1 Order #PB137X from the National Science Teachers Association, 1840 Wilson Blvd.,Arlington, VA 22201-3000; 800-722-NSTA. The NSTA has collaborated with the Society of Protozoologists to produce a selective, reviewed collection of 28 investigations; microscopes are the only "specialty equipment" required. This manual shows how to present an entire "live" biology class (morphology, physiology, ecology, ethology, taxonomy - everything!) with protozoa. It's intended for grades 9 and up, but parts can be adapted for use as a middle school supplement. More information is available at the NSTA website at http://www.nsta.org.pubs. High school. RECOMMENDED
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
Conrad Perfett wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello, } I have just subscribed to the group and am asking if there are any } Amateur Microscopist on this list? From what I can see it is mainly } professional especially when the discussion is on the electron variety! } I am an amateur in the UK. If anyone knows of a list that is more for } Amateurs I would be grateful too. } } Also where is the best type of Environment to find an Amoeba! I can find } all the other types of protozoa (such as Paramecium and coleps) yet have } failed to come across this text book example! } } Thanks in advance } } Conrad Perfett } } ------------------------------------------------------------------------ } ----------- } "Any sufficiently advanced technology is indistinguishable from magic" } ----------------------------------------------------------- Arthur C } Clarke ----
Conrad,
Please see Microscopy UK site at:http://www.microscopy-uk.org.uk/indexnew.html.
Henrik
-- Henrik Kaker SEM-EDS Laboratory, Metal Ravne d.o.o. Koroska c.14, 2390 Ravne, Slovenia Tel: +386-602-21-131, Fax: +386-602-20-436 SEM-EDS Laboratory Web Site http://www2.arnes.si/guest/sgszmera1/index.html Microscopy Vendors Database http://www.kaker.com/mvd/vendors.html Kaker.Com http://www.kaker.com
One of our group just did the drying routine with HMDS and we found lots of crystals during SEM examination. The crystals look either like christmas tree branches or like small pyramids. Since we have previously done the procedure with no problems we were wondering what went wrong. Anyone have any ideas?
} ---------- } From: Black, Cary (CK) } Sent: Tuesday, June 24, 1997 7:53 AM } To: microscopy-at-Sparc5.Microscopy.Com } Subject: MBX probe parts and pieces } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
A number of people have written me privately concerning my recent posting about university competition with private labs. Some of their thoughts (and my comments on them) are: } "the university (and I) need the money" -- true, but I think this is short-sighted. While your unit certainly benefits in the short run from the added income (and you are personally able to continue in your soft-money position), the long-term outlook is cloudy. In particular, such activities weaken the case for government support to university research. So, the downward spiral continues.
} Don, you're just looking out for your own selfish interests. -- true, but so is the individual who says "_I_ need the money from my university's contract services". Now consider that the owner of an analytical lab typically has _all_ of his or her personal assets (house, car, life savings) at risk. Put yourself in her position and see whether you'd feel differently. I'm not trying to change the system. I'm just looking for enforcement of existing regulations. In particular, I don't want to see my tax dollars go to support organizations that undermine my ability to earn the money to pay the taxes.
} I read Notice 91 and I'm sure it leaves some pretty wide loopholes, with room for interpretation. -- I intend to post the text of notice 91 on my web site (and I'll announce that here when I accomplish that). In the meantime, I beg to differ. Notice 91 is dated March 11, 1983. The cover letter, signed by Edward A. Knapp, director of the NSF (at that time), includes the following sentence: "It is contrary to the NSF's intent for grantees to use NSF-supported research instrumentation or facilities to provide services for a fee in direct competition with private companies that provide equivalent services." I feel that one's interpretation of notice 91 is a worthwhile test of ethical judgment. If you feel compelled to rationalize your participation in offering analytical services by saying "well, I'm not competing _directly_", then you may be engaging in "situational ethics" (bending the rules to fit the situation).
} My equipment and facilities weren't funded by the NSF, so notice 91 doesn't apply. -- true. In this case, you'll want to become familiar with OMB Circular A-110. Section __.34b reads: "The recipient shall not use equipment acquired with Federal funds to provide services to non-Federal outside organizations for a fee that is less than private companies charge for equivalent services, unless specifically authorized by Federal statute, for as long as the Federal Government retains an interest in the equipment."
respectfully addressed to the Microscopy community by
Don Chernoff, President
Advanced Surface Microscopy, Inc. E-Mail: asm-at-indy.net 6009 KNYGHTON RD. Voice: 317-251-1364 INDIANAPOLIS IN 46220 Toll free: 800-374-8557 (in USA) web: http://www.a1.com/asm Fax: 317-254-8690 (If you experience difficulty in accessing our website, note that the web address uses numeral "1" in "a1")
Dear All, Thanks for all the replies! I have been swamped and I mean this in the best possible sense of the word!! For those Amateurs interested, I have the following equipment:
Wedmore Microscope from Brunel Microscopes with bulb illumination complete (and a nice wooden case!) Objectives: x4, x10, x40, x63 Eyepieces: x5, x10, x12.5 thus giving a magnification range of x20 - x787.5 Camera attachment + adapter ring for my old Praktica SLR Camera (MTL 50) I also have Methyl blue general dye and the protozoa solution which is a must for slowing the little critters down!
I would be interested in what camera equipment other Amateurs use. I find the focusing screen on the camera makes focusing indistinct especially at higher powers of magnification. Would an SLR with a clear screen be better? I have been told there are cameras with an interchangeable screen but they are very expensive and so would like to enlightened as to how others go about photomicrography!
As to my background I am a programmer by day and although I am a fan of computer and associated technology, its nice to get away from them for my hobby! I was always interested in biology and related subjects and was thinking of becoming a biologist when at school but went into electronics etc instead!
Many thanks,
Conrad
------------------------------------------------------------------------ ----------- "Any sufficiently advanced technology is indistinguishable from magic" ----------------------------------------------------------- Arthur C Clarke ----
Dear All, Thanks for all the replies! I have been swamped and I mean this in the best possible sense of the word!! For those Amateurs interested, I have the following equipment:
Wedmore Microscope from Brunel Microscopes with bulb illumination complete (and a nice wooden case!) Objectives: x4, x10, x40, x63 Eyepieces: x5, x10, x12.5 thus giving a magnification range of x20 - x787.5 Camera attachment + adapter ring for my old Praktica SLR Camera (MTL 50) I also have Methyl blue general dye and the protozoa solution which is a must for slowing the little critters down!
I would be interested in what camera equipment other Amateurs use. I find the focusing screen on the camera makes focusing indistinct especially at higher powers of magnification. Would an SLR with a clear screen be better? I have been told there are cameras with an interchangeable screen but they are very expensive and so would like to enlightened as to how others go about photomicrography!
As to my background I am a programmer by day and although I am a fan of computer and associated technology, its nice to get away from them for my hobby! I was always interested in biology and related subjects and was thinking of becoming a biologist when at school but went into electronics etc instead!
Many thanks,
Conrad
------------------------------------------------------------------------ ----------- "Any sufficiently advanced technology is indistinguishable from magic" ----------------------------------------------------------- Arthur C Clarke ----
I copy this from a book of Olga Flint, Food Microscopy:
If you stain with aqueous iodine viewed brightfield and between crossed polars: Raw, amylose-containig starches -} blue, birefringent Raw, amylopectin starches-} Reddish, birefringent Chemically modified starches-} yellow or brown, birefringent Pre-gelatinized, i.e., precooked starch powders-} Reddish, swollen particles, which may contain blue-stained granules (not birefringent) Dextrins-} Blue-purple or reddish particles which quickly disperse Proteins in cryosections and powders-} yellow
I was taken aback by Conrad Perfett's enthusiasm about microscopy. His comments reminded my of when I was a boy and eagerly looked for the microscopic creatures swimming in the swamp in back of my house. What a thrill it was to find some single-celled wriggler with whipping flagella among the flotsam of decaying plants. My father, a high school biology and ecology teacher for a quarter of a century, could usually identify the animal, sometimes down to the species. Cool! Thanks Dad.
I work in an industrial lab and it is easy to lose that excitement in the midst of the latest crisis or long term project. Sometimes it takes a fresh voice like Mr. Perfett's to bring the thrill of discovery back. Thanks Conrad.
Harry Crossman
------------------------------------------------ Opinions or statements expressed herein, rational or otherwise, do not necessarily reflect those of my employer.
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 Phone: (508) 750-1717 E-mail: crossman-at-osi.sylvania.com
Our web sites: www.sylvania.com www.siemens.com --
} Fellow microscopists.......Thought I'd forward this response to my query on Cameca MBX bargraph tubes....................I ordered 2.
Cary
} } } } Just went through the search for a replacement Bar Graph for my MBX. } } } Source: Dale Electronics } } } 402-563-6286 (ask for Lynn) } } } Part Number: PBG-12201 } } } Cost: about $110. } } } } } I Just ordered 2. } } } } Cheers and happy probing } } } } } } Cary } } Dow Chemical } } }
Not that I disagree with your thesis, but I would interpret the section cited below as encouraging price-fixing. Significant differences exist between "academic" and private facilities. If the differences didn't exist, private facilities could not survive. While I don't believe (and I would hope this is true of everyone on the list) that academic facilities should use equipment funded by federal agencies to make a profit (or even to recover costs), a fair comparison of rates between the two organizations might lead one to believe that the advantages offered by the commercial groups are not worth the extra cost. What the customer pays for, in my estimation, are: convenience (i.e., access), detailed and professional reports, and skilled operation. Academic facilites can fall down in the last two categories. In the absence of a careful evaluation by the customer before the work is started, the academic unit will probably be a better deal because the extra dollars paid for the commercial services only assure better performance in the latter categories. If I were in charge of getting samples analyzed for some corportation, and I wanted to marry cost-effectiveness and "advancement of knowledge", I would go to commercial sources for my routine analyses (for consistency, speed, and quality reports) and send my "tricky" stuff to an academic "collaborator" (for new perspectives and state-of-the-art equipment). Rob Palmer Univ Tenn
} } My equipment and facilities weren't funded by the NSF, so notice 91 } doesn't apply. } -- true. In this case, you'll want to become familiar with OMB Circular } A-110. Section __.34b reads: } "The recipient shall not use equipment acquired with Federal funds to } provide services to non-Federal outside organizations for a fee that is } less than private companies charge for equivalent services, unless } specifically authorized by Federal statute, for as long as the Federal } Government retains an interest in the equipment." } } respectfully addressed to the Microscopy community by } } Don Chernoff, President
Please excuse if this is a second post, but my email here at work is often unreliable, and I don't think my initial posting made it to the listserver. So, here goes the question for a second time!
We have some polymer blends (ethylene-vinyl acetate and polyethylene) which most likely have blend domains of the PE in the EVA. We think we have seen these by SEM. I am looking for a method to selectively stain one phase (the EVA I assume) to confirm our domain theory.
Table 4.2 in Sawyer & Grubb's "Polymer Microscopy" lists some stains for esters, including EVA. These stains include phosphotungstic acid and methanolic NaOH. The text in that section discusses phosphotungstic acid, but not as applied to EVA; it fails to mention the methanolic NaOH at all.
Anyone have knowledge/experience with these stains? Many "thank-you's" will go to those who help out the domain analysis.
TIA,
Jim Passmore Cryovac North America
james.passmore-at-corp.wrgrace.com (often unreliable) jimpsm-at-aol.com (frustrating, but usually works)
I have been asked about my transcription of NSF Important Notice 91, which I had made available on the Web. We have just suffered from a hacker, so the server is offline until we re-secure it, but within the next couple of days, the text will be accessible again at http://18.82.0.42/nsf.in91/in91.html
Tony Garratt-Reed
**************************************************** **************************************************** ** ** ** Anthony J. Garratt-Reed ** ** Room 13-1027 ** ** Center for Materials Science and Engineering ** ** Massachusetts Institute of Technology ** ** 77 Massachusetts Avenue ** ** Cambridge, Massachsetts 02139-4307 ** ** U. S. A. ** ** ** ** Phone: 617-253-4622 ** ** Fax: 617-258-5286 or 617-258-6478 ** ** ** **************************************************** ****************************************************
I can only endorse what Henrik said about the Microscopy -UK site. It is one of the best I have seen and even has an on-line magazine 'Micscape' which is archived. I am sure that they have done one or two practical articles on protozoa but I notice that someone is doing a series on pondwater and will be covering protozoa in the near future.
Malcolm Haswell Electron Microscope Unit University of Sunderland UK ----------
Conrad Perfett wrote: } } Hello, } I have just subscribed to the group and am asking if there are any } Amateur Microscopist on this list? From what I can see it is mainly } professional especially when the discussion is on the electron variety! } I am an amateur in the UK. If anyone knows of a list that is more for } Amateurs I would be grateful too. } } Also where is the best type of Environment to find an Amoeba! I can find } all the other types of protozoa (such as Paramecium and coleps) yet have } failed to come across this text book example! } } Thanks in advance } } Conrad Perfett } } ------------------------------------------------------------------------ } "Any sufficiently advanced technology is indistinguishable from magic" } ----------------------------------------------------------- Arthur C Clarke ----
Conrad,
Please see Microscopy UK site at:http://www.microscopy-uk.org.uk/indexnew.html.
Henrik -- Henrik Kaker SEM-EDS Laboratory, Metal Ravne d.o.o. Koroska c.14, 2390 Ravne, Slovenia Tel: +386-602-21-131, Fax: +386-602-20-436 SEM-EDS Laboratory Web Site http://www2.arnes.si/guest/sgszmera1/index.html Microscopy Vendors Database http://www.kaker.com/mvd/vendors.html Kaker.Com http://www.kaker.com
Thank you very much for the informative replies on osmium precipitates.
Yours, Nuria Cortadellas Unitat de Microscopia de Transmissio Serveis Cientifico-Tecnics Universitat de Barcelona C/ Sole i Sabaris, 1-3 08028 Barcelona Tel: +34 3 402 13 52 Fax: +34 3 402 13 98 E-mail: nuriac-at-giga.sct.ub.es
} One of our group just did the drying routine with HMDS and we found lots } of crystals during SEM examination. The crystals look either like } christmas tree branches or like small pyramids. Since we have previously } done the procedure with no problems we were wondering what went wrong. } Anyone have any ideas?
Residual buffer salts. Wrinse (dilute) out the buffer prior to HMDS drying.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 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 ####################################################################
Has anyone coupled Cy 5 to membrane probes? I am trying to use a probe that will light up all phagosomal membranes, are some choices better? Molecular Probes does not use dyes in the infrared range, and that's what I need. Any and all advice is greatly appreciated. Has anyone coupled Cy5 to any gram negative bacilli, and maintained viability? Any tips? Again thanks for the help.
Anyone aware of any restrictions on equipment purchased through state or corporate donations?
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 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 ####################################################################
Some states have laws, but otherwise not. } } Anyone aware of any restrictions on equipment purchased through state or } corporate donations? } } #################################################################### } John J. Bozzola, Ph.D., Director } Center for Electron Microscopy } Neckers Building, Room 146 - B Wing } Southern Illinois University } Carbondale, IL 62901-4402 } 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 } #################################################################### } } } } ******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
If you just want a nonspecific membrane probe that fluoresces like Cy5, Molecular probes has Bodipy 665/676, cat # B-3932. It is completely hydrophobic, not amphipathic like DiI or the labeled phospholipids. It will label external membranes (and phagosomes too, I'm sure) but I can't tell you whether it will label internal membranes like mitochondria that don't exchange with the labeled plasma membrane. You can use a stock solution of a few mg/ml in DMSO; I haven't tried alcoholic stocks but they may work also. The price would be much cheaper than conjugating Cy5 yourself.
If you do find a source of Cy5 conjugated to lipids, please let me know!
Has anyone coupled Cy 5 to membrane probes? I am trying to use a probe that will light up all phagosomal membranes, are some choices better? Molecular Probes does not use dyes in the infrared range, and that's what I need. Any and all advice is greatly appreciated. Has anyone coupled Cy5 to any gram negative bacilli, and maintained viability? Any tips? Again thanks for the help.
A colleague is seeking an individual or lab experienced in autoradiography techniques (preferably with brain tissue) for some advice and potential contract work at the light and/or EM level. The tissue will be perfused rat brain containing the labelled compound directed toward cell membrane incorporation. A major goal is to determine whether the compound is specificically incorporated into nerve membranes. Currently things are in the planning stage, he has the C-14 labelled compound already in the animals and will have results on the extent and gross localization of incorporation in about two weeks. Whole brain lysates from a preliminary study indicates about 6000 cpm present (quenching effects are not known).
Specific areas he needs advice on: What will be the best fix, embedding combination for ultra-thin plastic, light and em sections etc. He would also be looking at hippocampal, cortex and proabaly one other area of the brain. The major question is how to do this to get the best localization at the ultra structural level in the neurons. He can have brains ready to go in about 2 weeks and he wants results by early September for publication and presentation at Neuroscience. Of course, whoever does it also gets their name on the paper.
If any experts out there can provide tips, tricks, pitfalls or are willing to actually collaborate on this project please contact:
Dr. Mark Clarke at mclarke-at-medics.jsc.nasa.gov
thanks
Edward J. Basgall, PhD The Pennsylvania State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618
We have been trying to prepare some specimens of unstained plasmid DNA on thin (approx. 3nm) carbon films but have been experiencing some difficulties in getting the DNA to stick properly. We have been trying to follow your procedure (Microbeam Analysis Vol. 2, 1993, pp. 69-79) as closely as possible. I was wondering how critical the glow-discharge of the films is? Also, do you have any other tips/suggestions that are not mentioned in the paper.
Any advice would be greatly appreciated.
Best regards,
Richard Leapman NIH, Bethesda (301) 496-2599 E-mail: leapman-at-helix.nih.gov
Soory about this, but I listed an outdated server address in my previous post. Please use the one listed below.
ed basgall ____________________________________________________________________ Hello fellow microscopists,
A colleague is seeking an individual or lab experienced in autoradiography techniques (preferably with brain tissue) for some advice and potential contract work at the light and/or EM level. The tissue will be perfused rat brain containing the labelled compound directed toward cell membrane incorporation. A major goal is to determine whether the compound is specificically incorporated into nerve membranes. Currently things are in the planning stage, he has the C-14 labelled compound already in the animals and will have results on the extent and gross localization of incorporation in about two weeks. Whole brain lysates from a preliminary study indicates about 6000 cpm present (quenching effects are not known).
Specific areas he needs advice on: What will be the best fix, embedding combination for ultra-thin plastic, light and em sections etc. He would also be looking at hippocampal, cortex and proabaly one other area of the brain. The major question is how to do this to get the best localization at the ultra structural level in the neurons. He can have brains ready to go in about 2 weeks and he wants results by early September for publication and presentation at Neuroscience. Of course, whoever does it also gets their name on the paper.
If any experts out there can provide tips, tricks, pitfalls or are willing to actually collaborate on this project please contact:
Dr. Mark Clarke at mclarke-at-sdmail.jsc.nasa.gov
thanks
Edward J. Basgall, PhD The Pennsylvania State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618
Here at the University of Delaware, we have an ISI WB-6 SEM that is going to be scrapped. We will be retaining the mechanical pump, the Polaroid film back, and possibly the ion pump assembly: with those exceptions, the entire machine, or any part of it, is available to anyone who will pick it up. We have both the chamber/HV unit and the console unit. This machine has not been run up for more than a year, but was operating when last used. Everything has been carefully labeled, disconnected, and tied up: the machine could be put back the way it was (nothing's been "ripped out"). We also have a small (scant) collection of manuals, and a box of schematics & drawings (annoted in Japanese). The machine has tungsten filament assemblies; we have a very few spare filaments. Also a handful of spare stubs. The University will not pack or ship. Anything wanted must be picked up by 4:30 PM EDT WED 02 JUL 97. Handcart, freight elevator, loading dock, and ramp down to ground level are all available. E-mail wieland-at-me.udel.edu for directions & arrangements. Sorry for the short notice, but I was not given the schedule & the order to remove it until yesterday.
Robert Wieland wieland-at-me.udel.edu You can't go faster than light, you can't get colder than absolute zero, and you can't help somebody by not telling them the truth.
You might also try staining the PE following the method of Hodge and Bassett (J. Mat. Sci. 12, (1977) 2065-2075. The method involves immersing the sample in chlorosulphonic acid at 60 C for a few hrs. ( they give a range of 3-27 hrs) in sealed containers. This is followed by soaking in uranyl acetate for 3 hrs. CHeck the reference for more details.
We got good results with extended chain PE fibers, I do not know for sure if it will work with your samples.
Please excuse if this is a second post, but my email here at work is often unreliable, and I don't think my initial posting made it to the listserver. So, here goes the question for a second time!
We have some polymer blends (ethylene-vinyl acetate and polyethylene) which most likely have blend domains of the PE in the EVA. We think we have seen these by SEM. I am looking for a method to selectively stain one phase (the EVA I assume) to confirm our domain theory.
Table 4.2 in Sawyer & Grubb's "Polymer Microscopy" lists some stains for esters, including EVA. These stains include phosphotungstic acid and methanolic NaOH. The text in that section discusses phosphotungstic acid, but not as applied to EVA; it fails to mention the methanolic NaOH at all.
Anyone have knowledge/experience with these stains? Many "thank-you's" will go to those who help out the domain analysis.
TIA,
Jim Passmore Cryovac North America
james.passmore-at-corp.wrgrace.com (often unreliable) jimpsm-at-aol.com (frustrating, but usually works)
Jordan Marti wrote: =========================================== So, I would appreciate it if Universities that accept contract work (on a per sample basis) for TEM of polymers would send me their address, contact name and tel. no. =========================================== I would respectfully question whether the above represents proper use of this listserver.
At least here in the USA there is a line that is drawn between what does and does not constitute appropriate use of the facilities of a university for commercial customers. While we might as reasonable people quibble about just where a line should be drawn, I would think that at least for the most part, we could all agree that there is a line somewhere. After all, a university that is organized as a nonprofit tax-exempt educational organization should not be competing in the commercial marketplace with for- profit tax-paying laboratories.
The best attempt yet at determining just where to draw that line, was NSF Important Notice 91. In a nut shell, at least the NSF, for the first time, recognized that it is quite inappropriate for any NSF grantee institution to be using NSF funded instrumentation in ways that are competitive with for- profit tax-paying firms offering substantially similar kinds of services. There is no prohibition against doing work for private firms so long as the results contribute to educational objectives, that is, the work is basic and fundamental in nature, suitable for inclusion in a student's thesis and the results would be intended for prompt publication. However work outside of those specifications is not permitted.
The request for universities that could do "contract work (on a per sample basis)" surely sounds to me like a call to university providers to make proposals that would be in direct competition with for-profit commercial firms and if the instrumentation was NSF-funded, then it is an open invitation for a university to violate its own agreements with NSF.
Further, with regard to Important Notice 91 being, as Gene Taylor says, "only" a guidline, again, maybe he should have a chat with his own Chancellor. The University of Maryland on an annual basis "signs off" on a special certification (as do all NSF grantee institutions) that they are in compliance with NSF Important Notice 91. So any thought that these are some lose set of suggestions, that can be followed or not followed in some discretionary way is pure nonsense. If this is the way some labs at the U. of Maryland are being operated, then they should be investigated by NSF. If other laboratories are certifying statements that are not correct, then the Office of Inspector General of the NSF should be taking a look at them as well.
The United States has no shortage of competent for-profit tax-paying laboratory organizations to do this kind of work. The only complaint is that such laboratories have higher charges than universities. But that should not be all that surprising given the fact that a university may have its instrumentation provided as part of a grant and enjoys a whole list of other advantages that flow to it because of its nonprofit tax-exempt and educational status.
So while I have my own vested interest in seeing that this type of work is done in the for-profit sector and not in the university sector, there is an element of fairness here, not to mention the legality of it, and it is for that reason that I respectfully suggest that such an advertisement for would -be university providers is not appropriate.
Disclaimer: Structure Probe, Inc. has been offering "contract work (on a per sample basis) for TEM of polymers". We have been an outspoken critic of universities that commercialize their facilities and compete unfairly, some would say illegally, with the private sector.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI Structure Probe, Inc. FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Since you've got Sawyer's and Grubb's book on polymer microscopy, you can check page 97, part 4.4.2.3 and the reference 99 which describes a method of pretreatment (alkaline saponification) before staining EVA with osmium tetroxyde.
Regards,
============================================================= Dr. Antoine Ghanem SOLVAY Research and Technology Rue de Ransbeek 310 1120 Brussels Belgium Tel. (32)(2)2643422 Fax. (32)(2)2642055 e-mail : Antoine.Ghanem-at-solvay.com =============================================================
how can I get in touch with US firm called Plasma Sciences Inc?
.attila(L)
Attila L. Toth ------------------------------------------ MTA MFKI Research Institute for Technical Physics of the Hungarian Academy of Sciences H-1325 Budapest POB 76 tel: (36.1) 169-2100 x 226 fax:(36.1) 169-8037 ------------------------------------------ email: tothal-at-mufi.hu (EUDORA:=CDrj =E9kesen!) ------------------------------------------
Hi all again, I now have a collection of photographs from my microscope! They are encouraging and have convinced me to invest in a camera with a clear focusing screen (such as the Olympus OM2 I have heard about from various sources) since more often than not they are just out of focus! The ones that are in focus are whetting my appetite though. I mean if I can get a sharp picture of one of these critters I may just get a small poster size photo done of it!
I also have a tiny microscope that is labelled 'BIJOU' no doubt due to its diminutive size! It is metal with removeable objectives and magnification range x100 - x300. It is not much taller than a coffee mug yet it is of much better quality than the cheap plastic toy microscopes. Has anyone any ideas of who manufactured it? I guess it is difficult without a photo of it, but maybe someone out there may know!
Conrad
------------------------------------------------------------------------ ----------- "Any sufficiently advanced technology is indistinguishable from magic" ----------------------------------------------------------- Arthur C Clarke ----
Just a general question to help me clear something up. I am sure this question has been asked a million times, but I see different answers from different sources.
ie what is the maximum theoretical magnification of a light microscope and what is the maximum USEABLE magnification? I have been told that around x1000 is the maximum useable, which makes me wonder why even with my microscope you can go up to x1350!
Following the maxim that most beginners buy microscopes with too high a magnification I decided that a good selection of low power objectives was the thing to have on mine which is why I have the x4 objective allowing me to go down to x20 which is useful to do a general 'search' when hunting the creatures down! I confess that I too have fallen into the trap of buying a 'toy microscope' whith too high a magnication which obviously cannot be used especially with lenses that are plastic and not adjusted for aberations!
Conrad.
------------------------------------------------------------------------ ----------- "Any sufficiently advanced technology is indistinguishable from magic" ----------------------------------------------------------- Arthur C Clarke ----
Are the small CCD cameras that can be purchased from electronics catalogues (such as MAPLIN for people in UK!) any good to build a video setup or are they not suitable? If they are, what additional equipment would you need to plug them into a TV? At first sight they seem financially within reach, but I guess the cost will probably escalate with additional bits and pieces!
Conrad
------------------------------------------------------------------------ ----------- "Any sufficiently advanced technology is indistinguishable from magic" ----------------------------------------------------------- Arthur C Clarke ----
For your PE/PVAc blends, you might have more success trying the Reading etching technique with permanganic reagent. For a first look, see my personal home page (URL below) and somewhere you will find a hyperlink _etching_ which should start you off, and give you the necessary references. On word of warning, the 1979 reference in SUPERSEDED and you should follow the 1982 recipe.
If you have any more questions, please feel free to contact me at any time. The staining technique was actually originated by Kanig, but we applied it to a variety of polymer systems so do look at the reference suggested by Jordi Marti.
And MOLTAS GRACIES / MUCHAS GRACIAS Jordi for bringing the technique to the attention of the Microscopy Newsgroup.
Best regards,
+------------------------------------------------------------------------+ | 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 | +------------------------------------------------------------------------+
Toth Attila wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear Colleagues, } } how can I get in touch with US firm called Plasma Sciences Inc? } } .attila(L) } } Attila L. Toth } ------------------------------------------ } MTA MFKI } Research Institute for Technical Physics } of the Hungarian Academy of Sciences } H-1325 Budapest POB 76 } tel: (36.1) 169-2100 x 226 } fax:(36.1) 169-8037 } ------------------------------------------ } email: tothal-at-mufi.hu (EUDORA:Írj ékesen!) } ------------------------------------------
Here is short information about Plasma Sciences, Inc.
Plasma Sciences, Inc. 7200 A Telegraph Sq. Dr. Lorton, VA 22079 USA Tel: 703 550 7888 Fax: 703 339 9860
-- Henrik Kaker SEM-EDS Laboratory, Metal Ravne d.o.o. Koroska c.14, 2390 Ravne, Slovenia Tel: +386-602-21-131, Fax: +386-602-20-436 SEM-EDS Laboratory Web Site http://www2.arnes.si/guest/sgszmera1/index.html Microscopy Vendors Database http://www.kaker.com/mvd/vendors.html Kaker.Com http://www.kaker.com
dear all im wondring if some one can give me a explication concerning EXELFS. exacetly about extraction of this signal. so in general they use a polynimial function to describe atomic distribution, i want to knowe why they use this than a power function who generaly used to remove a background.
Plasma Sciences is out of business; their products are available through:
South Bay Technologies East Coast Office 4019 S 16th St Arlington, VA 22204
phone & fax (703) 486-7999 ask for Steve Collins
Best regards, Steven Slap ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
Is there a methode to label negatively charge size results from carboxyl = groups and positive charge size results from amino groups at membran = layers like S-layer. The resolution I want to get are about 3nm. Due to = the fact that I also investigate additionally to S-Layer microtubuli = structure which are very sensitive to pH variation all labeling = techniques should be work at pH =3D 7 and if possible in solution.
Remo Kirsch "kirsch-at-tmfs.mpgfk.tu-dresden.de"
Technical University of Dresden / Germany Dept. Material research
Conrad Perfett wrote: } } I now have a collection of photographs from my microscope! They are } encouraging and have convinced me to invest in a camera with a clear } focusing screen (such as the Olympus OM2 I have heard about from various } sources) since more often than not they are just out of focus!
I use a Nikon F3 HP and an "M" focusing screen. Nikon F3's are becoming fairly inexpensive on the used camera market as Nikon owners move up to the newer automatic models. The "M" screen is clear with hatch marks for measuring scale. Also, the Nikon has a lock up mirror for less vibration, a feature not available on many other cameras.
Even with this effort to reduce vibration, I mount the camera on a copy stand. The camera is connected to the microscope, but it is supported by the copy stand. Vibration is, hopefully, thus transferred to the table and not the microscope. I also turn off all fans in the house before taking photographs.
As for focusing, the brain is a marvelous thing. It quickly adapts to things that are out-of-focus by making them appear in-focus to the eye. You cannot stare at an oject for any length of time to get it in focus. You have to quickly look at the object to see if it is in focus. If not, make a quick adjustment, and then look away for a short time. Look again, make another adjustment, and repeat the process. If you keep staring at the object while trying to get it in focus, after awhile the brain will remember the best image, no matter what is actually taking place in the microscope/camera combination.
These are only a couple of things that will affect photo results, and I've not even mentioned optics. If these things go bad a once, a photo of an object, appearing in-focus to the viewer, will be terribly blurred when the photograph is retrieved from the processor. The unevenness of the results will be difficult to identify until the user makes every effort to reduce vibration and errors of the brain.
I'd love to hear more on this subject. Thanks to Conrad for bringing it up.
I am sure this one will bring a flood of answers but the important thing is resolution (closely followed by contrast) ie an image must be sharp and different bits distinguishable. If you can achieve 0.2 micron resolution from a microscope and your eye can resolve 0.2 mm then 1000x would seem like a good limit.
In practice of course it's not just the magnification of the objective that is important (eg 100x) because eyepiece lenses may be 10x, 16x or something else. Also if you produce photographs then you can enlarge them and you may use a different lens from the eyepiece in the camera path. If this isn't bad enough then the quoted magnification may vary by at least 1 or 2% and you would need to use calibration specimens to check it.
The simplest way to check resolution would be to use Ernst Abbe formula: resolution - 0.61 x wavelength/NA the 0.61 constant may vary in other formulae and depends on how easy it is to distinguish two points wavelength of green light is about 0.5 micrometres NA (numerical aperture) of oil immersion lens can be 1.25 or a bit higher then resolution in the above formula = 0.244 micrometres You could try checking the NA of your lens and comparing it to this.
Obviously resolution can be improved by using shorter wavelengths or objective lenses with higher numeric apertures (usually printed on the lens somewhere). But there is a limit to the light that you can see (you can get UV microscopes, X-ray and electron microscopes using shorter wavelengths) and the numeric aperture is a simple product of the refractive index(n) and the sine of the semi-angle of the lens (sine alpha) and won't get much better than about 1.5. So microscopes may vary a bit.
If you want pictures much bigger than 1,000x then you would do it photographically anyway and often the reason for doing this is to view posters from a distance or measure things more easily but remember that you will suffer from "empty magnification" because the detail will become more blurred as you enlarge more.
I am sorry if this has rambled a bit but it's the simplest way I could do this without diagrams. I am a biologist and I can understand the above so I hope it's what you wanted.
Malcolm Haswell Electron Microscope Unit University of Sunderland UK ----------
Just a general question to help me clear something up. I am sure this question has been asked a million times, but I see different answers from different sources.
ie what is the maximum theoretical magnification of a light microscope and what is the maximum USEABLE magnification? I have been told that around x1000 is the maximum useable, which makes me wonder why even with my microscope you can go up to x1350!
Following the maxim that most beginners buy microscopes with too high a magnification I decided that a good selection of low power objectives was the thing to have on mine which is why I have the x4 objective allowing me to go down to x20 which is useful to do a general 'search' when hunting the creatures down! I confess that I too have fallen into the trap of buying a 'toy microscope' whith too high a magnication which obviously cannot be used especially with lenses that are plastic and not adjusted for aberations!
Conrad.
------------------------------------------------------------------------ "Any sufficiently advanced technology is indistinguishable from magic" ----------------------------------------------------------- Arthur C Clarke ----
The question is not what is the maximum magnification, but rather what is the maximum resolution. The old maxim (without quibbling about how many decimal places to run the answer out to) is that the eye can discriminate two objects that are about 0.2 of mm apart (I use to be able to do this when I was younger) and the theoretical diffraction limited resolution is about 0.2 of a um. Thus any magnification above 1,000X is empty magnification- no new structures will be discerned but people like me with weakening eyes may be able to see the smaller structures better. There are now available means of slightly increasing the diffraction limited resolution (i.e. discriminating slightly smaller objects) and make higher magnification lens practical.
This is the simple answer. I would be happy to provide details about how to extend the resolution if you need them.
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-bgsm.edu * **************************************************************
On Thu, 26 Jun 1997, Conrad Perfett wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Just a general question to help me clear something up. I am sure this } question has been asked a million times, but I see different answers } from different sources. } } ie what is the maximum theoretical magnification of a light microscope } and what is the maximum USEABLE magnification? I have been told that } around x1000 is the maximum useable, which makes me wonder why even with } my microscope you can go up to x1350! } } Following the maxim that most beginners buy microscopes with too high a } magnification I decided that a good selection of low power objectives } was the thing to have on mine which is why I have the x4 objective } allowing me to go down to x20 which is useful to do a general 'search' } when hunting the creatures down! I confess that I too have fallen into } the trap of buying a 'toy microscope' whith too high a magnication which } obviously cannot be used especially with lenses that are plastic and not } adjusted for aberations! } } Conrad. } } ------------------------------------------------------------------------ } ----------- } "Any sufficiently advanced technology is indistinguishable from magic" } ----------------------------------------------------------- Arthur C } Clarke ---- } }
In response to the following two postings: ---------------------------------- From Atilla Toth: how can I get in touch with US firm called Plasma Sciences Inc? --------------------------------- From Steven Slap: } } } } Plasma Sciences is out of business; their products are available through:
South Bay Technologies } } East Coast Office } } 4019 S 16th St } } Arlington, VA 22204 ----------------------------------- I can report the following:
On February 21, 1997 there was an involuntary filing (Chapter 7 liquidation) in the Bankruptcy Court, Eastern District Virginia (Alexandria Division) by Plasma Sciences, Inc. It is Case No. 97-11289-SSM.
On May 16, 1997, in another court action, the bankruptcy was converted to Chapter 11 of the Bankruptcy laws.
On June 26, 1997 there will be a meeting of the creditors (a "341" meeting it is called) at 2:00 pm, in Alexandria, VA.
September 24, 1997 is the bar date for the filing of claims.
Debtor's Bankruptcy Counsel: Steve Allen Mandell, Esq. Ph: (703) 734-9622
Creditor's attorney: David C. Lynn, Esq. Ph: (202) 628-6800
I plan to attend the creditor's meeting, representing SPI Supplies, on June 26 (today) and will communicate privately with anyone wanting to know what transpired, assuming I would not be divulging anything that should be deemed to be confidential. Just make your request by e-mail and I will reply by e-mail since such information would not be of general interest to this group.
If anyone has need to contact Plasma Sciences, Inc. on matters relating to their Chapter 11 filing, in theory at least, the company continues to operate so the phone and FAX numbers should still be operating.
If Steven Slap is correct in that they really are "out of business", then I can only say that the entire microscopy community has suffered a setback, this was a good firm, run by good people and they had a history of making good reliable products.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Well this list is certainly keeping me busy with the information I am getting! Especially useful are the tips on photomicrography. The information on Numerical Aperture was extremely useful as well.
Another in my endless list of questions! Is there a fairly comprehensive book on protozoa which actually has photographs that anyone could recommend. The reason being is that drawings tend to include all the features present in the creature which are not so obvious under a microscope making identification difficult. Paramecium being a prime example. I am sure that what I have viewed are these creatures but I cannot see a gullet or the nucleus at all. I have seen what I assume are rotifers but again identification is difficult, but then thats half the fun! I only have a little observers book on pond life, which for its 2 pound price is excellent considering it seems to list more protozoa etc than books 5 or 6 times its price!
Conrad
------------------------------------------------------------------------ ----------- "Any sufficiently advanced technology is indistinguishable from magic" ----------------------------------------------------------- Arthur C Clarke ----
Conrad Perfett wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Are the small CCD cameras that can be purchased from electronics } catalogues (such as MAPLIN for people in UK!) any good to build a video } setup or are they not suitable? If they are, what additional equipment } would you need to plug them into a TV? At first sight they seem } financially within reach, but I guess the cost will probably escalate } with additional bits and pieces! } } Conrad } } ------------------------------------------------------------------------ } ----------- } "Any sufficiently advanced technology is indistinguishable from magic" } ----------------------------------------------------------- Arthur C } Clarke ---- Conrad, I have used these cameras in a number of applications including custom light microscopes. they are ok. Output format (NTSC) will go right into your VCR, monitor... If your not imaging to the ccd array, that is you are relaying the image with the optics that come on most of the cameras...well most of them will rarify an atmophere... Distortion is pretty bad but... for a few bucks more (now your up to ~$150.US) you can buy these cameras with a C-mount adaptor (or have one made) & buy nice optice. ($50-$150.US).
std discaimer: I don't know 'em, they don't send me checks.
The rule of thumb that I was taught was 1000x the numerical aperture of the objective is the maximum usable magnification from a optical light microscope. I think of it in terms of: The light coming from a specimen is information, the more information you can collect the better. The numerical aperture is the direct indicator of the percentage of the information that the objective can collect from the specimen as measured by the angle of the cone of light that an objective can accept from the specimen. On most objectives the NA is the number printed next to the magnification power on the side of the objective. People pay big bucks to get objectives with bigger numbers on the side.
If you end up with an final image that is greater than 1000x NA, it is termed empty mag, since it is just bigger without resolving any more detail. However, we do this all the time when we project transparency slides across the room.
Enjoy
Bob Morphology Core U of Washington Seattle, WA, USA
On Thu, 26 Jun 1997, Conrad Perfett wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Just a general question to help me clear something up. I am sure this } question has been asked a million times, but I see different answers } from different sources. } } ie what is the maximum theoretical magnification of a light microscope } and what is the maximum USEABLE magnification? I have been told that } around x1000 is the maximum useable, which makes me wonder why even with } my microscope you can go up to x1350! } } Following the maxim that most beginners buy microscopes with too high a } magnification I decided that a good selection of low power objectives } was the thing to have on mine which is why I have the x4 objective } allowing me to go down to x20 which is useful to do a general 'search' } when hunting the creatures down! I confess that I too have fallen into } the trap of buying a 'toy microscope' whith too high a magnication which } obviously cannot be used especially with lenses that are plastic and not } adjusted for aberations! } } Conrad. } } ------------------------------------------------------------------------ } ----------- } "Any sufficiently advanced technology is indistinguishable from magic" } ----------------------------------------------------------- Arthur C } Clarke ---- } }
We have a problem with weak fluorescence using BRDU (bromodeoxy- uridine) which goes to the DNA in replicating cells. They are treated in a standard manner using a formalin fix, with immunocytochemistry using a primary and a fluorescent secondary antibody. So far they have been cut as 1 -micron thick resin sections. Would thicker sections improve anything? (my thought is that this will only increase background). My colleague is wondering if it is worth pursuing gold labelling at EM level.
Thanks in advance - Keith Ryan Plymouth Marine Lab., UK
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Conrad -
Borrow one from a friend and try it! All home video cameras will "plug in". And the little ball-shaped Handycam will do video capture for your computer. The simplest mount is a camera tripod. You shouldn't mount the camera rigidly on the scope;a vibration-free connection between camera & microscope is essential. It's easy. Go to the hardware store and buy a short length of the slightly flexible black foam that's sold for insulating water pipes. Slip it over eyepiece & lens & experiment with length and alignment.
Here's another item from the Project MICRO bibliography that's relevant:
Discovery Scope, Inc. 1992 Activity Booklets. 6-15pp each, paperback, 5.5x8.5", $3.00 each. Discovery Scope, Inc., P.O.Box 607, Green Valley AZ 85622; 800-398-5404. Titles available in this series are: Investigating wetlands, Investigating arthropods, Investigating seashore life, Investigating termites, Investigating protozoans, and Macrophotography. They're well written but relatively expensive, considering their brevity; all emphasize the Discovery Scope and its accessories.. The photography booklet is particularly useful and is recommended for the teacher or advanced student who wants to do still or video photos. Middle school-adult.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
The signal from a 1 micron section should look very weak. Thicker sections will increase the signal as long as the antiboby can get access to the tissue via hydrophilic resin like LR white or etch the resin away. If you have a signal you can see at the light level on 1 micron thick sections, the signal at the EM level should be great!
Bob Morphology Core
On Thu, 26 Jun 1997, Keith Ryan wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } This is on behalf of a colleague: } } We have a problem with weak fluorescence using BRDU (bromodeoxy- } uridine) which goes to the DNA in replicating cells. They are treated in a } standard manner using a formalin fix, with immunocytochemistry using a } primary and a fluorescent secondary antibody. So far they have been } cut as 1 -micron thick resin sections. Would thicker sections improve } anything? (my thought is that this will only increase background). My } colleague is wondering if it is worth pursuing gold labelling at EM level. } } Thanks in advance - Keith Ryan } Plymouth Marine Lab., UK } }
Position Available: The Laboratory for Ultrastructural Research at the Shriners Hospital Research Unit in Portland, Oregon has an opening for a junior level electron microscopy technician (Research Technologist II). This research laboratory offers expertise to collaborative projects which primarily involves transmission electron microscopy, with emphasis on immunocytochemical protocols, rotary shadowing, and some scanning electron microscopy, fluorescence microscopy, and histology. The goal of this small, two person facility is to provide support for a variety of basic research projects in the field of connective tissue microarchitecture. The laboratory is well equipped and has a history of excellent productivity and adequate funding.
Qualifications: The successful applicant must be well versed in a wide variety of microscopic methods, with at least 1.5 years of electron microscopy experience. BS degree in Biology or Chemistry a minimum, MSA certification desirable. The preferred candidate must be proficient in sample preparation techniques for transmission and scanning electron microscopy including: dissection, embedding, ultrathin sectioning, critical point drying, vacuum evaporation and photographic technique. Operating knowledge of transmission and scanning electron microscopes as well as light microscopes for brightfield, phase contrast and fluorescence imaging is essential. Experience in histology, cryo EM, immuno-EM methodology, microwave processing technique, confocal microscopy and image analysis software desirable. The applicant must have excellent communicative skills and the ability to work well with a variety of personalities. Must work independently with a high degree of efficiency.
Duties: All aspects of specimen preparation of a variety of biological samples for TEM, SEM, and histology. All aspects of photography. Accurate record keeping and flawless computer entry of data for future retrieval. Operation of transmission and scanning electron microscopes, light microscopes and related laboratory equipment. Laboratory maintenance including general housekeeping, ordering supplies, record keeping, and solution maintenance. Must be efficient, self motivated, and work independently under general supervision, exercising judgment in day to day assignments.
The position is full time with an attractive benefits package and is available August 31, 1997. Current salary range for this position begins at $28,184 per annum. Send CV, statement of future goals, salary requirements and names of three references to:
Douglas R. Keene Electron Microscopy Facility Portland Shrine Research Unit Shriners Hospital for Children 3101 S.W. Sam Jackson Park Road Portland, Oregon 97201 FAX: 503-221-3451 ---------------------- Doug Keene DRK-at-shcc.org
} We have some polymer blends (ethylene-vinyl acetate and polyethylene) which } most likely have blend domains of the PE in the EVA. We think we have } seen these by } SEM. I am looking for a method to selectively stain one phase (the EVA } I assume) to } confirm our domain theory.
Our user today is working with polymers, so I asked him. He thinks that RuO4 will stain the two polymers differentially, and that he doesn't know anything that will be completely selective. He says the relevant referrence for RuO4 is by Trent from the early 80's. Good luck. Yours, Bill Tivol
We used iodine to stain the EVA for PLM work contrast since the PE wouldn= 't absorb it. This would probably work for SEM especially with an EDX syste= m.
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
There are two ways to answer your question concerning the useful magnification of an optical microscope. The rule of thumb used by most microscopists is that the maximum useful mag is 1000 times the numerical aperture (page 38 of Chamot and Mason's Handbook of Chemical Microscopy). An immersion lens can have a numerical aperture around 1.3 which gives 1300 times as the highest magnification. For a dry objective of 40 times magnification with N.A. 0.75 the useful magnification is 750 times. BUT, that rule of thumb is just that - a general rule. I measure particles using an image analyser and the effective magnification to the computer monitor is better than 2000 times and I use that for the best precision of measurements and since I'm well over forty and my personal optical system is degrading. I find the additional mag useful to say the least!
A better way of thinking of this issue is to consider the resolving power. This is often stated as the smallest feature which can be resolved using the specific optical system. It depends on the wavelength of light used and on the numerical aperture of the objective. The equation is 1.2 times the wavelength of light divided by 2 times the numerical aperture (page 13 of Chamot and Mason). That yields a limit of resolution of approximately 0.2 micrometers for the immersion objective I described and about 0.5 micrometers for the dry objective. So my 2000 times computer monitor magnification may help my aging eyes but I get no more information than at 750 times mag.
Jim: I agree with Bill's suggestion. Vapour staining with RuO4 will work. It will stain the polymers differentially. If you know the ratio of the two polymers in the blend it will be obvious otherwise you may have to experiment staining a blend of the two of a known ratio with the one predominant in the blend. This seems to be the best approach. However, you will have to section the blend at a temperature -120C or so due to PE. Then stain sections in RuO4 vapours for 10-15 minutes. If you need more information you may give me a call. Good Luck!
Vin Berry GE Plastics 304 863 7528 {vinod.berry-at-gep.ge.com} } ---------- } From: PBGVM1/FORWARDR } Sent: Thursday, June 26, 1997 1:47 PM } To: Berry, Vinod (GEP) } Subject: FORWARDED NOTE } } To: P1PTP37 --MSMAIL1 } } From: Your PROFS Id } Subject: FORWARDED NOTE } *******WARNING********* This note has been forwarded to you from your } Profs } userid. YOU MUST USE THE "FORWARD" ICON TO RESPOND TO THE ORIGINAL } SENDER -- } YOU CANNOT USE THE "REPLY" ICON. } ====================================================================== } === } ====================================================================== } === } Received: from thomas.ge.com by GE1VM.SCHDY.GE.COM (IBM VM SMTP V2R2) } with TCP; } Thu, 26 Jun 97 13:25:03 EST } Received: from ns.ge.com (ns.ge.com |192.35.39.24|) by thomas.ge.com } (8.8.4/8.7 } .5) with ESMTP id NAA22473; Thu, 26 Jun 1997 13:25:29 -0400 (EDT) } Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com } |206.69.208.10|) by } ns.ge.com (8.8.4/8.7.3) with SMTP id NAA06150; Thu, 26 Jun 1997 } 13:24:57 -0400 } (EDT) } Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com } (8.6.11/8.6.11) id M } AA10792 for dist-Microscopy; Thu, 26 Jun 1997 12:14:20 -0500 } Received: from gatekeeper.wadsworth.org (gate1.wadsworth.org } |199.184.16.5|) by } Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id MAA10789 for } {microscopy-at-s } parc5.microscopy.com} ; Thu, 26 Jun 1997 12:14:18 -0500 } Received: (from uucp-at-localhost) by gatekeeper.wadsworth.org } (8.7.1/8.7.1) id NA } A05908; Thu, 26 Jun 1997 13:11:45 -0400 (EDT) } Received: from newton.wadsworth.org(172.16.1.6) by gatekeeper via smap } (V1.3) } |id sma005903; Thu Jun 26 13:11:31 1997 } Received: (from tivol-at-localhost) by newton.wadsworth.org } (8.8.6.Beta3/8.7.1) id } NAA18355; Thu, 26 Jun 1997 13:13:14 -0400 (EDT) } From: William Tivol {tivol-at-wadsworth.org} } Message-Id: {199706261713.NAA18355-at-newton.wadsworth.org} } Subject: Re: EM-polymer staining } In-Reply-To: {C5F7CF9A-at-MHS} from "James.Passmore-at-corp.wrgrace.com" at } "Jun 25, } 97 08:29:59 am" } To: James.Passmore-at-corp.wrgrace.com } Date: Thu, 26 Jun 1997 13:13:14 -0400 (EDT) } Cc: microscopy-at-sparc5.microscopy.com } X-Mailer: ELM |version 2.4ME+ PL28 (25)| } MIME-Version: 1.0 } Content-Type: text/plain; charset=US-ASCII } Content-Transfer-Encoding: 7bit } Errors-to: Microscopy-request-at-sparc5.microscopy.com } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } ---------------------------------------------------------------------- } -. } } } Dear Jim, } } } We have some polymer blends (ethylene-vinyl acetate and } polyethylene) which } } most likely have blend domains of the PE in the EVA. We think we } have } } seen these by } } SEM. I am looking for a method to selectively stain one phase (the } EVA } } I assume) to } } confirm our domain theory. } } |Our user today is working with polymers, so I asked him. He thinks } that RuO4 will stain the two polymers differentially, and that he } doesn't } know anything that will be completely selective. He says the relevant } referrence for RuO4 is by Trent from the early 80's. Good luck. } } ||||Yours, } ||||Bill Tivol } }
-- ******************************************************************** Pat Kingman Continuum Mechanics and Analysis Team Immediate plans: Code jock....Computer hawk....Cybercadet...
Although I have not done biological microscopy, my general experience in both light & electron optical microscopy is that in many if not most practical situations, the character of the specimen and the quality of preparation limit the resolution long before the limits of the microscope are reached. Resolving the limit with a test specimen is one thing; resolving the same level of detail in a practical sample is often another. A large proportion of the important information in practical microscopy is discerned at a fraction of the maximum magnification.
Also, the sample may intrinsically lack contrast. (My favorite people are the ones who show up with a trashed-up piece of high-strength martensitic steel and are disgruntled because the micrographs don't look "like this"..."this" being somebody's carefully selected award-winning micros of titanium twins or beta brass.)
With light microscopy in particular, the depth of field at high mags is often the limiting factor in how high you can go. Many novices who sit at a research metallograph for the first time rush to crank the magnification to the max and are disappointed. As well, you can feel like fool after spending hours looking for the wrong thing..which you discover after 10 seconds with a low-power stereo microscope!
In message {c=US%a=_%p=Historical_Colle%l=IT3NT-970626102235Z- 5419-at-it3nt.pasttimes.com} , Conrad Perfett {ConradPerfett-at-pasttimes.com} writes } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Don't go for one of the small pcb mounted cameras they sell with a lens attached, the distortion is terrible.
Actually their C mount lenses are not too wonderful (no iris!) but you can probably workt without a lens if you remove the eyepiece from the microscope and make an adaptor (I had a 35mm camera mount on my Zenit which worked that way) The Objective should focus straight onto the ccd.
The output is the same as a camcorder. They won't plug directly into the TV arial - that needs a UHF signal. If the TV has a SCART connector you can use that (you can buy an adaptor at Dixon's or Curry's) but they will always plug into a VCR's video input which can then be connected to the TV. Or you can video your experiments and really get one up on your relatives holiday video.
-- Anthony James Bentley Surface Data Scientific Instrumentation and Software Web site http:\\www.surface.demon.co.uk
Hot darn! A conversation about light microscopy, resolution and magnification, which are some of my favorite subjects, so here comes my two cents:
Yes, the rule of thumb is magnification is equal to 1000 times N.A. under ideal conditions. I suggest a better rule of thumb is 850 times N.A.
So why do microscopes come with magnification greater then 1000X? The 1000*N.A. pertains to resolution of a small distance between two points. If the details we want to image are greater than this distance you can increase the magnification above 1000*N.A. and still get a good image. The fine detail will be fuzzy. Also, some time what we want is a large N.A. so we use low power oculars. N.A and magnification are linked, but we don't have to use the max magnification for every purpose.
Thanks for time and good luck! Frank ---------------------------------------------------------------------------- -------
These opinions are mine alone and have no relationship to my employer. Thank you.
Frank Karl fskarl-at-goodyear.com Goodyear Tire & Rubber Co. Voice: 330-796-7818 Analytical Services Dept. 415B Fax: 330-796-3304 142 Goodyear Blvd. Akron, OH 44305 USA
They that give up essential liberty to obtain a little temporary safety deserve neither liberty or safety. Benjamin Franklin
Another perspective on this thread... after working in and managing a "cost center" for materials analysis at a major university for several years, several points (most of which have been addressed) are clear 1) universities are doing TEM/SEM/AFM analysis for outside clients, and yes this is in direct competition with commercial labs. 2) most facilities which do this know of and try to comply with NSF Important Notice 91, and set their price schedules accordingly. and only conduct such business to defray their maintenance contracts on their equipment, not to "earn income" (ie. profit)
which leads to my point, the NSF/NIH is willing to grant funds to acquire equipment, but rarely is their any funding for maintenance/upgrades for that equipment. researchers are spending millions of tax dollars for equipment which requires up to $100k/year in service contracts (lets call it insurance) but often find out too late that they cannot use equip. money for maint., or choose to get all the bells and whistles on the equip. and neglect to budget for maint.
so the catch 22 begins...
should we allow this equip. to be used without insurance? or should the ins. be included in the purchase price? or should we let the uninsured equip. sit idle, after spending all the cash to get it until we find a way to support it? or do we form a non-profit govt. subsidized entity to raise the necessary money to pay for the service contracts?
we tried various combinations of all these ideas, and still have many more questions than answers.
I do BRDU with peroxidase on formalin fixed sections of mouse CNS and find that some sort of antigen retrieval is necessary to get good labeling. The formalin fixation is the culprit. Either switch to acid alcohol fix or treat the formalin-fixed sections with trypsin or hot buffer. E-mail me directly if you need more information.
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 e-mail: mcauliff-at-umdnj.edu ***************************************************************
A discussion has been going on in our lab regarding storage of valuable samples in a vacuum desiccator. Question: If the samples are stored in a plastic storage box, then put under vacuum, will the plastic from the boxes eventually coat everything (especially the samples) in the vacuum desiccator? Thanks in advance - Katharine
Katharine Kato SETI Institute 239-14 NASA Ames Research Center Moffett Field CA 94035-1000 ph# 415-604-5218 fax# 415-604-1088
} ------------------------------------------------------------------------ There is a little children's book I made with Robert Allen and Sean Morrison calle "the Amoeba". It was published by Coward, McCann & Geoghegan, Inc.NY in 1971. It has interesting photomicrographs of Amoebas and a few other protozoans. You might enjoy it. Regards, Nina Allen } Well this list is certainly keeping me busy with the information I am } getting! Especially useful are the tips on photomicrography. The } information on Numerical Aperture was extremely useful as well. } } Another in my endless list of questions! Is there a fairly comprehensive } book on protozoa which actually has photographs that anyone could } recommend. The reason being is that drawings tend to include all the } features present in the creature which are not so obvious under a } microscope making identification difficult. Paramecium being a prime } example. I am sure that what I have viewed are these creatures but I } cannot see a gullet or the nucleus at all. I have seen what I assume are } rotifers but again identification is difficult, but then thats half the } fun! I only have a little observers book on pond life, which for its 2 } pound price is excellent considering it seems to list more protozoa etc } than books 5 or 6 times its price! } } } Conrad } } ------------------------------------------------------------------------ } ----------- } "Any sufficiently advanced technology is indistinguishable from magic" } ----------------------------------------------------------- Arthur C } Clarke ----
Nina Stromgren Allen Professor, Department of Botany Box 7612 North Carolina State University Raleigh, NC 27695-7612 Phone: 919-515-8382 (Office), 515-3525 (Lab), 515-2727 (Department Secretary) Fax: 919-515-3436
In order to keep up with our continued growth. EDAX International is looking for an Applications Specialist for our Mahwah, NJ location. The primary functions of this position would be providing applications support and performing demonstrations of our x-ray microanalysis product lines.
The applicant should have good working knowledge and experience with microanalysis systems, and should possess excellent communication, presentation, and "people" skills.
Resumes may be sent to: Jim Nowak EDAX International 91 McKee Drive Mahwah, NJ 07430 (201)-529-6229 phone (201) 529-3156 fax
Someone in cyberspace recomended Loctite 460 adhesive to use as a means to attach samples to the Tripod Polisher's pyrex pedestle. We tried dozens of products but this one has excellent work time and adhesion similar to a product which we used until it's work time decreased as noticed on our last order. Loctite 460 is a good thing. Happy Thinning!
Stanley John Klepeis
P.S. Whover sent us the recomendation for Loctite 460, Thank's.
Katharine Kato wrote: ============================================= A discussion has been going on in our lab regarding storage of valuable samples in a vacuum desiccator. Question: If the samples are stored in a plastic storage box, then put under vacuum, will the plastic from the boxes eventually coat everything (especially the samples) in the vacuum desiccator ? ============================================= We have supplied to SEM users for some number of years (since about 1976) plastic storage boxes for SEM mounts. Other suppliers offer different boxes utilizing different plastics, but polymers in general don't "evaporate" and "redeposit" somewhere else, at least not the kinds of molecular weights being used for the boxes. The same would be true for the base plates that hold the mounts.
However, the plasticizer in some of the box bottoms is another story. You want to be careful NOT to put in even for a short term, "reactive" samples such as might have been given the O-T-O osmium tetroxide procedure. Now in that case, the osmium will definitely react, but primarily with the plasticizer in the base plate, and the end result is a fine somewhat feathery deposition onto your mounted samples. And I have never found anyone who has been able to resurrect such samples for further examination.
So if your samples are not of the "reactive" type, I can see no reason why you would have any problems, except where noted above.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
What kind of plastic is that storage box made of? Most plastics don't have or need plasticizers. An exception is PVC, which is frequently plasticized, mostly to give it a "soft touch" (like in car dashboards or fake leather). Probably the maker of a sample box wouldn't plasticize the PVC for that application (they certainly wouldn't want to put much in since the box has to be rigid), but you may want to stay away from PVC just in case.
Disclaimer: These are my personal opinions and not those of my employer. My employer's parent company is involved in myriad plastics businesses. ____________________
} A discussion has been going on in our lab regarding storage of valuable } samples in a vacuum desiccator. Question: If the samples are stored in a } plastic storage box, then put under vacuum, will the plastic from the boxes } eventually coat everything (especially the samples) in the vacuum } desiccator? } Thanks in advance - Katharine
} Katharine Kato } SETI Institute } 239-14 NASA Ames Research Center } Moffett Field CA 94035-1000 } ph# 415-604-5218 fax# 415-604-1088
Well its a small world, Last night as I was selling my computer to someone and he noticed that I had a microscope and it turned out that he was a retired scientist who worked at a hospital in Bristol (here in UK) using microscopes for years. He was quite fascinated to find someone who had microscopy as a hobby and was saying that you don't meet often meet people who have a 'scientific' hobby that often! I guess this is so true in this world of Video games etc. (My excuse is that I am useless at video games anyway!) Of course a fascinating conversation inevitably followed.
As someone on here remarked about themselves, it was a case of someone doing microscopy for a living entering the world of computers while I am a programmer (therefore in the world of computers!) entering the world of microscopy!
One thing that never ceases to amaze me is how toy manufacturers treat microscopes like cars ie high spec is the thing to have regardless of how useful one may find it! A lot of the microscopes you see in Toys 'R' Us have magnifications around 750x to x1000!! which given the plastic lenses is really conning young kids don't you think?
Conrad Perfett
------------------------------------------------------------------------ ----------- "Any sufficiently advanced technology is indistinguishable from magic" ----------------------------------------------------------- Arthur C Clarke ----
You need to define your standard method for BrdU. We routinely do frozen sections at 3 um with excellent results. Are you certain the amount of BrdU you are injecting(?)/treating your animal(?)/cells with is adequate? Our standard procedure is to fix in 4% paraformaldehyde for 1 hr, prepare for 3 um frozen sections and prior to the ICC we treat the tissue for 1/2 hr - 45 min. with 2N HCl. This gives us excellent labeling of dividing cells found in the teleost retina. We have found that a Cy3 fluorescent conjugate works well for us. Do you have an absolute need to embed in resin? This may be introducing a problem with pentration and antigen/antibody binding. We also have found that in our hands any glutaraldehyde fixation significantly reduces or eliminates BrdU ICC.
Linda Barthel Research Associate II Department of Anatomy and Cell Biology University of Michigan lab (313) 764-7476 fax (313) 763-1166 barthel-at-umich.edu
Katherine, I have been storing my thin film samples in vacuum desiccators and haven't found any problem. In general, most of the plastics which are used for these applications do not erode when put in vacuum unless it reacts with sample or temperature is raised. Hence, there shouldn't be any problem in storing samples in plastic desiccators.
senthil
//// ___|--00_____________________________________________________________________ C ^ S.Senthil Nathan \ ~/ Vacuum and Thin Films Lab., Dept. of Instrumentation, {} {} Indian Institute of Science, Bangalore, India - 560 012 e-mail:sen-at-isu.iisc.ernet.in URL : http://isu.iisc.ernet.in/~sen/ voice: +80 3092349 fax: 91 80 3345135
On Thu, 26 Jun 1997, Katharine Kato wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } A discussion has been going on in our lab regarding storage of valuable } samples in a vacuum desiccator. Question: If the samples are stored in a } plastic storage box, then put under vacuum, will the plastic from the boxes } eventually coat everything (especially the samples) in the vacuum } desiccator? } Thanks in advance - Katharine } } Katharine Kato } SETI Institute } 239-14 NASA Ames Research Center } Moffett Field CA 94035-1000 } ph# 415-604-5218 fax# 415-604-1088 } }
Toy microscopes quote areal not linear magnifications so these actually have top mag of ~30x. Good microscopes quote a mag on the eyepiece and on the objective. I wonder which sort your 'Bijou' is? In UK the Royal Microscopy Society founded by Amateurs over 150 years ago is now running a campaign called AMFES A Microscope For Every School to encourage the use in primary school of a well made basic microscope rather than a cheap over specified one. Up to date and accurate information rather than my vague recollections from
Royal Microscopical Society 37/38 St Clements Oxford OX4 1AJ, UK They are also on the internet. } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Well its a small world, } Last night as I was selling my computer to someone and he noticed } that I had a microscope and it turned out that he was a retired } scientist who worked at a hospital in Bristol (here in UK) using } microscopes for years. He was quite fascinated to find someone who had } microscopy as a hobby and was saying that you don't meet often meet } people who have a 'scientific' hobby that often! I guess this is so true } in this world of Video games etc. (My excuse is that I am useless at } video games anyway!) Of course a fascinating conversation inevitably } followed. } } As someone on here remarked about themselves, it was a case of someone } doing microscopy for a living entering the world of computers while I am } a programmer (therefore in the world of computers!) entering the world } of microscopy! } } One thing that never ceases to amaze me is how toy manufacturers treat } microscopes like cars ie high spec is the thing to have regardless of } how useful one may find it! A lot of the microscopes you see in Toys 'R' } Us have magnifications around 750x to x1000!! which given the plastic } lenses is really conning young kids don't you think? } } Conrad Perfett } } ------------------------------------------------------------------------ } ----------- } "Any sufficiently advanced technology is indistinguishable from magic" } ----------------------------------------------------------- Arthur C } Clarke ---- } }
} One thing that never ceases to amaze me is how toy manufacturers treat } microscopes like cars ie high spec is the thing to have regardless of } how useful one may find it! A lot of the microscopes you see in Toys 'R' } Us have magnifications around 750x to x1000!! which given the plastic } lenses is really conning young kids don't you think? } } Conrad Perfett } Conrad makes an excellent point here, and one that I think the MSA educational people might help with. We all want kids and primary schools to use microscopes, not just for our own self-interest, but because of their general value in science education. But most 'scopes for children are not only cheaply made, but as Conrad notes, over-spec'ed. And I agree with him that the claims made for these toy 'scopes are deceptive, and are likely to disappoint both the child and the parent. Then no more microscopy, no science interest, the kid grows up a bitter, twisted Artist, and ends with a Performance Art piece wrapping the planet in plastic, ... the possiblities are horrible.
What is needed is a good, 5X-100X binocular stereoscope for about US$100 (less would be better--$50, say). This would allow children (and teachers) to examine insects, many protozoans and much of pond life, minerals, fractures, salt crystals, and of course, the ubiquitous "e". Such a scope could be made with good quality optics that would give a clear, undistored image, unlike the cheap toys sold in stores.
I have seen an instrument sold through Science News and maybe the Edmund catalog that is something like this, but not binocular, and of a more limited mag range. (I don't know about the optics.)
A question for the MSA officials: given the arguements in favor of microscopy in primary schools, and as a home hobby, how about MSA designing or endorsing such a microscope? That official imprimateur might make parents more willing to spend $50 or $100 for a 'scope instead of $10 or $25, as it would be an assurance of quality. MSA educational materials could be included. If Tasco or Edmund (or ... ) could be talked into being the merchant for this, MSA could even get some bucks for this.
Phil
} Sic Hoc Legere Scis Nimium Eruditionis Habes { Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217) 355-1143 oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
Item Subject: What is maximum magnication of light microscope?
Conrad,
If you want a good source of used microscopes and other "stuff" you might try the following:
Martin Microscopes Easley, SC Phone: 803-859-2688 803-242-3424 fax 803-859-3332
I got a great little microscope from them some years ago that my kids enjoy immensely. They frequently have used "school" microscopes that they resell and more objectives than they even know that they have.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Just a general question to help me clear something up. I am sure this question has been asked a million times, but I see different answers from different sources.
ie what is the maximum theoretical magnification of a light microscope and what is the maximum USEABLE magnification? I have been told that around x1000 is the maximum useable, which makes me wonder why even with my microscope you can go up to x1350!
Following the maxim that most beginners buy microscopes with too high a magnification I decided that a good selection of low power objectives was the thing to have on mine which is why I have the x4 objective allowing me to go down to x20 which is useful to do a general 'search' when hunting the creatures down! I confess that I too have fallen into the trap of buying a 'toy microscope' whith too high a magnication which obviously cannot be used especially with lenses that are plastic and not adjusted for aberations!
Conrad.
------------------------------------------------------------------------ ----------- "Any sufficiently advanced technology is indistinguishable from magic" ----------------------------------------------------------- Arthur C Clarke ----
} } One thing that never ceases to amaze me is how toy manufacturers treat } } microscopes like cars ie high spec is the thing to have regardless of } } how useful one may find it! A lot of the microscopes you see in Toys 'R' } } Us have magnifications around 750x to x1000!! which given the plastic } } lenses is really conning young kids don't you think? } } } } Conrad Perfett } } } Conrad makes an excellent point here, and one that I think the MSA } educational people might help with. We all want kids and primary schools to } use microscopes, not just for our own self-interest, but because of their } general value in science education. But most 'scopes for children are not } only cheaply made, but as Conrad notes, over-spec'ed. And I agree with him } that the claims made for these toy 'scopes are deceptive, and are likely to } disappoint both the child and the parent. Then no more microscopy, no } science interest, the kid grows up a bitter, twisted Artist, and ends with } a Performance Art piece wrapping the planet in plastic, ... the } possiblities are horrible.
Phil Oshel
you are right but I believe the same can be said about any kind of industry or business branch. there is a big mistake between the price of things and their value, as said recently the late Captain Cousteau. I know this has nothing to do with microscopy but it is worth thinking a bit more deeply sometimes about errouneous behaviours...
Many thanks for book references, Could people supply ISBN numbers if possible when quoting a book since here in UK it would make it easier when trying to locate books especially if they have similar names!
Conrad
------------------------------------------------------------------------ ----------- "Any sufficiently advanced technology is indistinguishable from magic" ----------------------------------------------------------- Arthur C Clarke ----
Phil Oshel Wrote: A question for the MSA officials: given the arguments in favor of microscopy in primary schools, and as a home hobby, how about MSA designing or endorsing such a microscope? That official imprimateur might make parents more willing to spend $50 or $100 for a 'scope instead of $10 or $25, as it would be an assurance of quality. MSA educational materials could be included. If Tasco or Edmund (or ... ) could be talked into being the merchant for this, MSA could even get some bucks for this. ----------------------------------------------------
I see a lot of value in this possibility. If MSA did endorse a good school / home educational scope for children and hobbies etc.
This could bring the MSA organization to the public attention, and become a more publicly known logo / organization.
This would also lead to possibilities such as future microscopists or laboratory scientists. Young people having become familiarized with MSA and it's resources may go into microscopy fields when they otherwise would not. Especially with good tools. I remember such a great disappointment in my toy scope as a youngster, I had expected so much more. Also some of the people exposed early on to the organization may end up being the bosses and administration that would ok or veto funding for microscopy related equipment etc etc. There are a lot of possibilities for a stronger future in doing things like this.
As a Medical Technologist, I see parallels between the public's perception of a Registered Nurse (RN) verses a Medical Technologist [MT(ASCP)]. Everyone understands, ( and most appreciate) what a Nurse does. Many do not understand or appreciate a Medical Technologist. ( perhaps because we pack needles for blood drawing sometimes) We are often asked if we went to a whole year of schooling ( try 5 years of college, 34 hours of week of scheduled class time -- all science). Med Techs are often called nurses. Often has been the lament that our recognition, saleries etc would have benefited from a more public persona. The same could be said for microscopists.
Lou Ann
Lou Ann Miller Center for Microscopy & Imaging College of Veterinary Medicine Dept. of Veterinary Biosciences University of Illinois Rm 1108 Basic Sciences Bld 2001 S Lincoln Ave. Urbana, Illinois 61802
Phone: 217-244-1566 email: lamiller-at-uiuc.edu
Center for Microscopy & Imaging Home page: http://www.cvm.uiuc.edu/MicImagLab/MicImagLab.html
Central States Microscopy Society: http://www.cvm.uiuc.edu/Homepages/LouAnnMiller/CSMS/csms.html
Personal Home Pages: http://www.cvm.uiuc.edu/Homepages/LouAnnMiller/LAM.html
We have an opportunity to purchase a small freeze dryer in the near future for materials applications (mostly aqueous polymers for GPC, Thermal and some microscopy work). I am looking for some feedback on the following possibilities: 1. Labconco Lyph-lock 4.5L This is a basic model with a 4.5 liter ice capacity, a -54 C condenser and comes in floor or bench models. Cost is ~$5.5K without a vacuum pump.
2. VirTis Benchtop 5-SL, -55 C This unit also has a -55 C condenser and has a 4 liter ice capacity. The advantages of this unit are a built in vacuum pump and shell bath for freezing samples can be included. Cost is $9160 with the pump and shell bath. The manifold is ~1K extra. Total cost is ~$11K.
3. VirTis FreezeMobile 5EL, -85 C This unit has a -85 C condenser, allowing operation with organic solvent systems, and requires 208 volt line. Unit is floor mounted on casters and has a 2.5 liter ice capacity. Total cost for this unit with vacuum pump, shell bath and manifold is ~$13K.
I am open to other suggestions as well. I am concerned about efficiency, ease of use and reliability of the units.
jeharper-at-amoco.com wrote: } } } Conrad, } } If you want a good source of used microscopes and other "stuff" you might try } the following: } } Martin Microscopes } Easley, SC } Phone: 803-859-2688 } 803-242-3424 } fax 803-859-3332
New Area Code is 864.
And Bob Martin is excellent. Tell him Eric Metzler said "Hi"
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Phil -
I disagree with your call for a BINOCULAR inspection/dissection scope, because 1) many children have difficulty with convergance, 2) the interocular spacing usually can't be set for young faces, 3) binocular is both more expensive and easy to misalign in rough classroom/home use, & 4) 10% of the population is ambliopic (including me!), with faulty binocular vision. There is an excellent 20x monocular available; the ultimate source is a Hong Kong distributor, and it's sold under a variety of brand names in the U.S for ~$80-90. I'll send a dealer list to anyone who sends me a SASE. The RMS already has an "approved" list (see my reply to Conrad in today's Email), which includes the scope described above. I'll be discussing the approval concept with MSA's Standards committee at the August meeting. Is anyone out there interested in serving on a scope review subcommittee? You, Phil? A MSA scope has been suggested, but the society (my opinion) shouldn't get into the business if the private sector has a good product at a fair price; the "approved" list is needed first. "Microscopic Explorations", the MSA-sponsored manual, will be published in the Lawrence Hall of Science GEMS (Great Explorations in Math & Science) series next spring. That's a top quality series which is sold by several major suppliers. Caroline
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } One thing that never ceases to amaze me is how toy manufacturers treat } microscopes like cars ie high spec is the thing to have regardless of } how useful one may find it! A lot of the microscopes you see in Toys 'R' } Us have magnifications around 750x to x1000!! which given the plastic } lenses is really conning young kids don't you think? } Conrad - It's worse than a con job; it's likely to turn kids off completely, since such junk is somewhere between frustrating and impossible to use. You can get a list of "Royal Microscopical Society approved" children's microscopes (currently 20x inspection/dissection scopes for the primary grades, 50x compound coming soon) from Juliet Dyson, coordinator of the RMS "A Microscope For Every School" program. Her address is Moor Gate Farm, Netherthong, Holmfirth, Huddersfield HD7 2UP, U.K. English addresses are charming, but I wish she had Email... Caroline
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
Bart Cannon wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello, } } Does anyone know of a source for bulk volumes of the blue luminescing } plastic scintillator material which is sometimes used in SEM electron } detectors? } } Thanks. } } Bart
Company NE America is producer of various scintillator materials: Here is short information from the Microscopy Vendors Database (http://www.kaker.com/mvd/vendors.html):
NE America Princeton Corporate Plaza 7 Deer Park Drive Monmouth Junction, NJ 08852 USA Tel: 201 329 1177 Fax: 201 329 2221 Full range of NIM modules, plastic scintillators, special liquid scintillators, crystal detectors of all types.
-- Henrik Kaker SEM-EDS Laboratory, Metal Ravne d.o.o. Koroska c.14, 2390 Ravne, Slovenia Tel: +386-602-21-131, Fax: +386-602-20-436 SEM-EDS Laboratory Web Site http://www2.arnes.si/guest/sgszmera1/index.html Microscopy Vendors Database http://www.kaker.com/mvd/vendors.html Kaker.Com http://www.kaker.com
Hi there, does anyone know of a commerc ially available Nomarski microscope with a near field attachment which can take 200 mm wafers? I am interested in looking at very small (tens of nm) features on wafer surfaces after they have been detected on a light scattering tool. The stage would have to be programmable to be able to go to the co-ordinate given by the light scattering tool. Any suggestions?
thanks
Lucio
Dr.Lucio Mule'Stagno MEMC Electronic Materials Inc University of Missouri -St.Louis Silicon Materials Research Group Physics Dept., 501 Pearl Dr., 8001 Natural Bridge rd., St.Peters, St.Louis MO 63376 MO 63121 tel: 314 279 5338 314 516 5931/3 fax: 314 279 5363 314 516 6152 email: lmulestagno-at-memc.com luciom-at-newton.umsl.edu
Hi Lynne, I would opt for the freeze dryer that can hold the lowest temp. If you are using aqueous suspensions you want to remain below the ice recrystalization point ( { -80C). Of the choices listed the last one seems the best. You might also want to consider vacuum cleanliness. If you do any subsequent chemical analysis can you tolerate pump oil contaminants? I would want a turbo or cryosorption pumped freeze dryer.
good luck
ed
Edward J. Basgall, PhD The Pennsylvania State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618
} We have an opportunity to purchase a small freeze dryer in the near } future for materials applications (mostly aqueous polymers for GPC, } Thermal and some microscopy work). I am looking for some feedback on } the following possibilities: } 1. Labconco Lyph-lock 4.5L } This is a basic model with a 4.5 liter ice capacity, a -54 C } condenser and comes in floor or bench models. Cost is ~$5.5K without } a vacuum pump. } } 2. VirTis Benchtop 5-SL, -55 C } This unit also has a -55 C condenser and has a 4 liter ice } capacity. The advantages of this unit are a built in vacuum pump and } shell bath for freezing samples can be included. Cost is $9160 with } the pump and shell bath. The manifold is ~1K extra. Total cost is } ~$11K. } } 3. VirTis FreezeMobile 5EL, -85 C } This unit has a -85 C condenser, allowing operation with organic } solvent systems, and requires 208 volt line. Unit is floor mounted on } casters and has a 2.5 liter ice capacity. Total cost for this unit } with vacuum pump, shell bath and manifold is ~$13K. } } I am open to other suggestions as well. I am concerned about } efficiency, ease of use and reliability of the units. } } Thanks in advance, } } Lynne Garone } GaroneL-at-Polaroid.com
Dear microscopists, Does anyone know of a reliable study relating color to the thickness of Silicon ( {20 micron)? I have been using one set of values, and then came across another set which was quite different, and I am now wondering which (if either) is correct. Thanks in advance for your time and help.
Mick Thomas Materials Science Center Cornell University
Tasco manufactures of 10X,20X and 30X stereomicroscope which sells at their Kent, Washington outlet store for $97.00. Its image is sharper and more chromatic aberration-free than that obtained by my B&L Stereozoom 7.
We are starting some in situ work with digoxigenin labeled probes. A lot of the kits and studies seem to use Alkaline Phosphatase labeled antibodies to detect the digoxigenin. I am curious why they seem to prefer Alk Phos over peroxidase coupled antibodies which are the more standard immunocytochemical choice. Anybody have any thoughts on this?
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
} Phil - } } I disagree with your call for a BINOCULAR inspection/dissection } scope, because 1) many children have difficulty with convergance, 2) the } interocular spacing usually can't be set for young faces, 3) binocular is } both more expensive and easy to misalign in rough classroom/home use, & 4) } 10% of the population is ambliopic (including me!), with faulty binocular } vision. There is an excellent 20x monocular available; the ultimate source } is a Hong Kong distributor, and it's sold under a variety of brand names in } the U.S for ~$80-90. I'll send a dealer list to anyone who sends me a SASE.
These are good points, but I would argue that there are equally valids arguments against monocular 'scopes. I have had numerous students in college classes who got headaches, or otherwise couln't cope with a monocular, but did well with a binoc. The obvious solution is to offer both.
I would argue against a fixed mag. 'scope. They have a limited usefulness, and usually cause more frustration that anything else. Whatever mag is choosen, it will either be too high or too low for the specimens kids want to look at.
} The RMS already has an "approved" list (see my reply to Conrad in } today's Email), which includes the scope described above. I'll be } discussing the approval concept with MSA's Standards committee at the } August meeting. Is anyone out there interested in serving on a scope } review subcommittee? You, Phil?
Yes, but given the situation implicite in my signature, this is not likely to be practical. Ask me again in a couple of months or so.
} A MSA scope has been suggested, but the society (my opinion) } shouldn't get into the business if the private sector has a good product at } a fair price; the "approved" list is needed first. "Microscopic } Explorations", the MSA-sponsored manual, will be published in the Lawrence } Hall of Science GEMS (Great Explorations in Math & Science) series next } spring. That's a top quality series which is sold by several major } suppliers. } Caroline } } Caroline Schooley } Educational Outreach Coordinator } Microscopy Society of America
I agree the Society shouldn't be in the business of making or selling microscopes, but there is too much expertise in the society membership not to be involved in designing them, or choosing one or more to endorse.
I also think that although it is very important to get good microscopes for kids and schools, any such program should include adult hobbyists. One reason (yes, of many) astronomy gets funding is the strong amateur astronomer community, which includes corporate and government people. Why shouldn't MSA--and other countries' societies--support and encourage microscopy amateurs? This can only be good for microscopy.
The manual sounds excellent. This is the sort of thing that could be included with a MSA endorsed/designed product.
Phil
} Sic Hoc Legere Scis Nimium Eruditionis Habes { Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217) 355-1143 oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
Hi Can anyone suggest what to use as a length standard with a 100x objective? I intend to print out an image of this standard when I print images of samples, in order to determine final magnification. I am a little concerned with the use of my micrometer slide (10 microns between graduations) because the width of the individual lines is significant compared to the distance being measured between lines. Also there is no tolerance stated for the separation between the lines. I can think of using calibrated latex beads suspended in water, but the refractive index difference between latex and the aqueous medium (or air) causes a lot of problems. Are there other standards I can use?
I have an outstanding reference scale made by John McCaffrey* of the National Research Council of Canada. He has shown the silicon ranging in thickness up to 10 microns using both daylight and a tungsten filament. It is really quite nice. It should be up on our web site soon, but hasn't made it there yet. I do have a digitized copy that I could probably attach to an e-mail message. Of course, I am a bit of a novice when it comes to that stuff so you may end up with a picture of my 2 year old son instead - not an altogether bad thing either! :-)
Anyway, I should have it up on the web site by next week and then you could download it and print it out on a nice color printer. We also plan to make them into mouse pads soon, if you or anyone else would like one when we have them available, please contact me directly via e-mail.
* As a matter of interest, John is also the developer of the MAG*I*CAL, the world's smallest ruler (according to the Guinness Book of World Records). The MAG*I*CAL is a TEM calibration sample which is used to perform all of the 3 major calibrations for a TEM: 1) Magnification at all magnification ranges; 2) Camera constant and; 3) Image diffraction pattern rotation calibrations. We will have these available at the MSA meeting in Cleveland. Sorry for the semi-commercial plug, but it really seemed to fit in with the posting.
David Henriks TEL: 800-728-2233 (toll-free in USA) South Bay Technology, Inc. 714-492-2600 1120 Via Callejon FAX: 714-492-1499 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.Best regards Message text written by Malcolm Thomas } Dear microscopists, Does anyone know of a reliable study relating color to the thickness of Silicon ( {20 micron)? I have been using one set of values, and then came across another set which was quite different, and I am now wondering which (if either) is correct. Thanks in advance for your time and help.
Mick Thomas Materials Science Center Cornell University {
My apologies to the list, but Robin's email isn't working or something.
Robin, please send me your current address. The one on the email you sent me malfunctions.
Phil
} Sic Hoc Legere Scis Nimium Eruditionis Habes { Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217) 355-1143 oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Marcello:
We have several customers who have used both our Model 650 Low Speed Diamond Wheel Saw and our Model 860 Diamond Band Saw for cutting titanium implants. The Model 650 is more precise and more gentle, but the Diamond Band Saw can cut faster and is less expensive.
Just thought I'd throw in a few options for you. If you'd like more detailed information, please contact me directly.
David Henriks TEL: 800-728-2233 (toll-free in USA) South Bay Technology, Inc. 714-492-2600 1120 Via Callejon FAX: 714-492-1499 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 "Marcelo Henrique Prado da Silva" } Hello,
I'd like to know how could I get histological specimens from titanium implants inserted into rabbit bone. The problem is cutting bone with metal. It seems that the suitable equipment is named Exact. How does it work? What about its price? Where can I get it?
On Jun 27, 3:36pm, Edward J. Basgall wrote: } Subject: Re: Recommendations on freeze dryers } .... } I would opt for the freeze dryer that can hold the lowest temp. If you are } using aqueous suspensions you want to remain below the ice recrystalization } point ( { -80C).
Just to remind everybody that the recrystallization of vitrified water has been directly observed to take place at a temp of at least 135 - 140 K, i.e. -138 C to -133 C. Lit: Dubochet and MacDowell 1981 Dubochet and Lepault 1984 see also: Bachmann and Mayer, 1987, Chapter 1, in: Cryotechniques in Biological EM (Steinbrecht and Zierold, ed) Springer Berlin
Dr. Reinhard Rachel {Reinhard.Rachel-at-biologie.uni-regensburg.de} Universitaet Regensburg, Lehrstuhl fuer Mikrobiologie D - 93040 Regensburg (Tel.: xx49-941-943-4534) http://www.biologie.uni-regensburg.de/Mikrobio/Stetter http://www.biologie.uni-regensburg.de/Mikrobio/Stetter/Gruppen/rachel.html
} Dear microscopists, } Does anyone know of a reliable study relating color to the thickness of } Silicon ( {20 micron)? I have been using one set of values, and then came } across another set which was quite different, and I am now wondering which } (if either) is correct. Thanks in advance for your time and help. } } Mick Thomas
Mick,
a bit more than a year ago Daniel Callahan posted the following, that I believe is what you are looking for. I suppose it is still available, though I haven't checked it out.
------------------ forwarded Good Day:
I have placed a picture of a flat-wedge of optically transmitting (100) silicon on a web page: the image has thickness scale bars from 0 to 10 micrometers. This isn't an ideal sample: for example, there are no interference fringes visible in the submicron regime. However, it should serve as a starting reference. Please let me know what you think. The image is at
http://www.owlnet.rice.edu:80/~dlc/silicon.html
I am considering making a higher angle wedge hoping to preserve more of the thin region and also taking an image under a Na lamp as advised by Ron Anderson. Other suggestions are welcome as would other images, particularly of other orientation.
Daniel L. Callahan Department of Mechanical Engg. and Materials Science Rice University dlc-at-owlnet.rice.edu _____________________ end
Yves MANIETTE Universitat de Barcelona Serveis Cientifico Tecnics Unitat ESCA TEM Carrer Lluis Sole i Sabaris, 1-3 E-08028 BARCELONA ESPANYA
To the microscopy list: The discussion concerning microscope resolution which then moved to toy microscopes has been quite interesting: Abbe formula, NA values, and use disappointment.
I did not read in any of these discussions the concept of the point spread function of the optics of the microscope in question. As I understand it, the point spread function is a measure of the quality of the optics. As an image passes through the optics of the instrument, it is convolved. The point spread function is a measure of the degree to which the image in degraded. Microscope manufacturers (toy and otherwise) do not share this number with buyers.
For some time now I have hoped to see a public domain or shareware deconvolution algorithm which could take care of digitally recorded microscope images. Commercial deconvolution software usually costs $6,000 or more. With the software, one can vastly improve the image quality of a microscope, especially less expensive microscopes. I have seen prices of software like Authorware come down in price from over $5,000 to less than $500. I have not seen the same movement in price reduction for deconvolution software.
Could someone comment on point spread functions or deconvolution algorithms?
Blystone in Texas
Robert V. Blystone, Ph.D. {RBLYSTON-at-Trinity.edu} Professor of Biology Trinity University San Antonio, Texas 78212 210.736-7243 210.736-7229 FAX
Hello everyone, I operate a Philips SEM 505 interfaced to a Tracor Northern 5500 energy = dispersive x-ray spectrometer (EDX). If anyone would like to share = their ideas/methods of interfacing the now almost ancient (but working = perfectly) DEC PDP 11 which is the main guts of the TN5500 I would love = to hear from you. My email address is roleka-at-octonline.com or tel 1-800-561-5551 ext 2304 during business hours (or leave voice = mail). Thanks in advance for responces Ron Oleka
{P} {FONT face=3DArial size=3D2} I operate a Philips SEM 505 interfaced to = a Tracor=20 Northern 5500 energy dispersive x-ray spectrometer (EDX). If anyone = would=20 like to share their ideas/methods of interfacing the now almost ancient = (but=20 working perfectly) DEC PDP 11 which is the main guts of the TN5500 I = would love=20 to hear from you. {/FONT}
{P} {FONT face=3DArial size=3D2} My email address is {A=20 href=3D"mailto:roleka-at-octonline.com"} roleka-at-octonline.com {/A} {/FONT}
{P} {FONT face=3DArial size=3D2} or tel 1-800-561-5551 ext 2304 during = business hours=20 (or leave voice mail). {/FONT}
{P} {FONT face=3DArial size=3D2} Thanks in advance for responces Ron=20 Oleka {/FONT} {/P}
This message might come under you get what you pay for. I will also try to KISS (Keep It Simple Stupid), as the original questioner was an amateur. I won't even get into Achromats, Fluorites, or Apochromats.
The quotes below are from POLARIZED LIGHT MICROSCOPY by McCrone, McCrone, and Delly. They say it a lot better and succinctly than I ever could.
Lenses have several types of aberrations which will cause the loss of detail unless corrected for. Toy microscopes are not.
Spherical aberration - "is especially apparent in lenses having sperical surfaces. Light paths near the center of the lens focus at different points compared to light paths near the periphery."
Chromatic aberration - "is caused by refractive index variations with wavelength (dispersion). Thus, a lens receiving wihte light from an object will form a blue image closest to the lens, a red one farthest away." This is what causes those color fringes in the toy microscopes.
Field curvature - "is a natural result of using lenses with curved surfaces. The image plane produced by such lenses will be curved. This kind of image occurs in microscopy unless plano (flat-field) objectives are used."
Now onto a point on resolution and Numerical Apertures (NA - a measure of the resolving power) which I have not seen covered. Lenses having an NA greater than 1.0 require that an immersion oil be used. That oil must be used both between the top of the cover slip and the objective *AND* between the condenser and the bottom of the slide. If one of these is missing the effective NA of the "system" will be that of the air space, theoretically 1.00.
To the amateurs - welcome! We need you to keep us fresh and excited in what we do.
Shalom from Jerusalem,
Azriel Gorski, Head Optical Microscopy Laboratory Division of Identification and Forensic Science Israel National Police
I've stirred a little interest in the product and a little commentary from my mailing about the performance of the "$97" stereo microscope.
The scope was purchased from:
Tasco Sales, Inc. 7818 So. 212th Street Kent, WA 980032
The price was really $97.99 + tax. Tasco sales' hours and months of operation are variable. The same scope appears to be available from numerous other sources including Edmund Scientific as their part A52,167 with a price of $199.00 in catalog N971. I have no sales association with any of the companies I mention.
In my mailing I stated that the "$97" scope's image was sharper and more chromatic aberration free than that of my B&L Stereozoom 7. It has been pointed out to me that this can not be true if my B&L is truly in good shape. My Stereozoom 7 is 12 years old and has never been serviced. Since no specs are published with the cheapie I wonder.... the $97 scope has fixed objectives and straight-line eyepieces.... is it possible that, though less ergonomic, it possess a less compromised light path? Or... am I just a poor observer?
My testimonial was based upon a side by side comparison of the two scopes at 30X using the same light source. The subject was a sparkling array stout to slender, colorless 1 mm quartz crystals studded with 0.2 mm light green diopside crystals. The cheapie's image was whiter, brighter and sharper than the B&L with less dazzle and fringing.
I would not prefer to use the $97 scope on a regular basis over my B&L. The cheapie's straight eyepieces make it less comfortable, its depth of focus and field of view might be a little smaller and zoom magnification is a dream compared to fumbling with the cheapie's loose objective lenses.
My business requires about an hour a day around the stereomicroscope. I could actually get my work done with only minor sacrifices using the $97 scope. This makes the scope more of a TOOL than a TOY to me, and at about 1/35th (?) the cost of the Stereozoom 7, a possible bargain for those on a budget and who require only short scope hours.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Azriel - Thank you for the excellent summary; you've extracted it from a good source. Now what we need are a few REALLY SIMPLE tests that can be used by parents, teachers (and maybe even school district purchasing agents!) to use before purchase. Not to select for research quality, but to eliminate junk. Would anyone like to make a suggestion? Caroline
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
{ {Now what we need are a few REALLY SIMPLE tests that can be used by parents, teachers (and maybe even school district purchasing agents!) to use before purchase. } }
Why not "suggest" they take a sample of what they will expect to be viewed? For example a selection - a mounted flie wing, pollen, a algal mount.... these slides are cheap and could be used to evaluate at different magnifications - cheaply.
another lurking amateur.... -- Jacklyn L. Ryrie Mid Kirk, Bower, Wick,
} Azriel - } Thank you for the excellent summary; you've extracted it from a } good source. Now what we need are a few REALLY SIMPLE tests that can be } used by parents, teachers (and maybe even school district purchasing } agents!) to use before purchase. Not to select for research quality, but } to eliminate junk. Would anyone like to make a suggestion? } Caroline } Caroline,
A finely ruled grid could be used to check for pincushion and barrel distortions, as well as flatness of field, spherical aberration, and field size. If mass-produced, this could be made cheaply. A plate with various sizes (for different mags) of tiny pinholes might be used to check for chromatic aberration--color fringes inside the hole. A slide of radiolarians or diatoms (centric would be best) could also be used for this. These latter would probably be preferable to pinholes, as they would be cheaper, but species with simple tests would need to be chosen.
Phil
} Sic Hoc Legere Scis Nimium Eruditionis Habes { Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217) 355-1143 oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
Dan Callahan, I and two others recently published a direct measure of the colors of silicon in transmission as a short note - MICRON, Vol. 27, No. 6, pp. 407- 411 (1996). If you email me off the listserver, I'll send you a reprint. The sample used was a 7 degree cleaved wedge of single crystal silicon. This unique sample was imaged optically, by SEM and by TEM, and the three sets of images compared. There is another study that has just been submitted that uses a 2 degree tripod polished silicon sample, where the colors are again illustrated and the interference fringes analysed. Again, if you are interested, please email me and I'll get the information to you. This is basically a thin films problem, where the tristimulus values and the position and intensity of the interference fringes can be calculated.
Cheers John McCaffrey ------------------------------------------------------------------------ ------ REPLY FROM: McCaffrey, John (IMS)
} Dear microscopists, } Does anyone know of a reliable study relating color to the thickness of } Silicon ( {20 micron)? I have been using one set of values, and then came } across another set which was quite different, and I am now wondering which } (if either) is correct. Thanks in advance for your time and help. } } Mick Thomas
Mick,
a bit more than a year ago Daniel Callahan posted the following, that I believe is what you are looking for. I suppose it is still available, though I haven't checked it out.
------------------ forwarded Good Day:
I have placed a picture of a flat-wedge of optically transmitting (100) silicon on a web page: the image has thickness scale bars from 0 to 10 micrometers. This isn't an ideal sample: for example, there are no interference fringes visible in the submicron regime. However, it should serve as a starting reference. Please let me know what you think. The image is at
http://www.owlnet.rice.edu:80/~dlc/silicon.html
I am considering making a higher angle wedge hoping to preserve more of the thin region and also taking an image under a Na lamp as advised by Ron Anderson. Other suggestions are welcome as would other images, particularly of other orientation.
Daniel L. Callahan Department of Mechanical Engg. and Materials Science Rice University dlc-at-owlnet.rice.edu _____________________ end
Yves MANIETTE Universitat de Barcelona Serveis Cientifico Tecnics Unitat ESCA TEM Carrer Lluis Sole i Sabaris, 1-3 E-08028 BARCELONA ESPANYA
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
These are good tests, but when I say REALLY SIMPLE, I mean any hardware or art supply store. Maybe plastic window screening for the grid and real pinholes in aluminum foil?
Caroline
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
} Dear microscopists, } Does anyone know of a reliable study relating color to the thickness of } Silicon ( {20 micron)? I have been using one set of values, and then came } across another set which was quite different, and I am now wondering which } (if either) is correct. Thanks in advance for your time and help. } } Mick Thomas } Materials Science Center } Cornell University
Don't know the answer to your problem but ....
My understanding is that the colour at various thicknesses is dependent on, among other things, the bandgap, which in turn is orientation dependent. So, 100 Si will have a different colour/thickness dependence compared with 111 or 110.
This might account for the different valuses you have seen.
I am looking for all types of images (light & EM) which deal with algae, especially sea weeds. I plan to teach a course in which I will compare and contrast vegetative and reproductive adaptations of algae with land plants. The emphasis will be at the light microscope level, but SEM and TEM images which deal with major aspects of form and function would be welcome.
I would like to embed and cut murine femurs which were treated in vivo with tetracycline, and hence must remain undecalcified. At present we are fixing in formalin or paraformaldehyde and embedding in methacrylate plastic. We are cutting transverse sections and quantifying the amount of bone formation between the 2 tetracycline labels using confocal microscopy. We are having trouble with the actual sectioning, as the bone cracks and breaks up. We are presently trying to cut them using a tungsten carbide knife at between 3 and 5 microns. Any ideas to make this tedious process easier would be greatly appreciated.
I regret to inform you that the subscribe/unsubscribe functions will be off-line for a few days. During a system reconfiguration I managed to corrupt some system files, and while backup's exist I cannot for the moment access them to restore the status quo.
The listserver should continue to function (I hope) but the subscriber list is in statis until I sort out the problem.
Hi Maya I don't know about the first, I've never had really good results from cleaning off coated grids, but as to the second; in the absence of a Glow Discharge apparatus you might get some result out of using a Zerostat Antistatic Pistol. It is not so reliable as Glow Discharge and takes some practice to obtain neutral charge grids.
They are still available as far as I know. We bought one recently for zapping our Balance after frustrating sessions of attempting to weigh out powders which were flying all over the bench. It cured that.
If you want to see if it works before buying, find yourself a Black Vinyl Record collector and borrow one. They often use them for discharging static on Discs before playing. I used one myself before going over to CD's many years ago. Regards Stephen Griffiths
} Just wondering about a couple of things. } First the different ways there are to clean grids that have a } film (2% parlodion), and a carbon coat, but no sample. } Second the best way to reduce or eliminate static on coated grids, } without using a glow discharge apparatus. } Thanks, } Maya Moody
Thanks for the address etc. I already have an excellent microscope from Brunel Microscopes (so if there is anyone connected with them here, you can be assured that I am happy!) I have heard various people say what they think would be an ideal started microscope and low magnifications. I would have thought a monocular with say x20 x50 and x100 would be a good start since x20 is ideal to search with and x100 enlarges the subject to a nice level of detail. I have been doing my first photomicrographs using this magnification.
Considering some 'toy' microscopes can cost up to 50UK pounds I think it would be better for them to junk all the extras in the dearer kits as this seems to be what you pay up to ?30 extra for, as the microscope itself doesn't seem all that different from that sold on its own! Another case of manufacturers enticing kids into thinking 'more' is better! Considering the base model of the microscope that I purchased cost just 79 UKpounds and is solid metal for a start, I am sure that they could at least put glass lenses in the cheap toy ones!
Conrad ------------------------------------------------------------------------ ----------- "Any sufficiently advanced technology is indistinguishable from magic" ----------------------------------------------------------- Arthur C Clarke ----
Last week I started an 'infusion' with some slightly composting grass that was left in a container after the lawn had been cut a while ago. It seems that the lazy ways are sometimes the best ways since there is now a nice collection of protozoa including the ubiquitous paramecium although I still haven't found an Amoeba! I guess I got the wrong impression from books that they are the most common protozoa. I also got a a look at the bacteria swarming away digesting the grass which is where having the higher powers of my microscope came in useful! This is where looking at a real live culture beats the hell out of pictures or even prepared slides! Its fascinating seeing how fast some of them actually move!
Conrad
------------------------------------------------------------------------ ----------- "Any sufficiently advanced technology is indistinguishable from magic" ----------------------------------------------------------- Arthur C Clarke ----
Related to the discussion on toy microscopes, I also have a pet hate in toy binoculars x2 or x3. I know they have to be cheap and plastic, but it irks me that a roughly $15 pair from a "reputable" toy manufacturer performs worse than a $3 set from Hong Kong. This is simply because the "reputable" ones have got a wrong design for the meniscus of the objective.
Next up in price one finds all these 21mm aperature x20 zooms from Tasco, etc. The aperture-magnification ratio is all wrong. One has to go to $200 or so to get a half decent pair. This will put the kids off astronomy, too!
Sorry for intruding a semi-relevant grouch, but on matters like this, I run under OScar in the Grouchputer, the world's most unfriendly computer from Sesame Street Systems:
Garbage In - Yes Please (delete that last word) Gargage Out - Well whad'ya expect!!!
+------------------------------------------------------------------------+ | 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 | +------------------------------------------------------------------------+
1. *Before* you go to buy/test a microscope - define what *you* want the microscope to do and in simple task testable terms.
2. If you can talk to/take someone who knows his way around microscopes, do so. Many professional microscopists will be willing to devote some time to "hook" newcomers on something they love.
3. Sit and *play* (no I did not say test, I said play, your students will) with the microscope. Take and examine samples that you plan to use from your decission in step No. 1. *Tune into your impressions*, if the microscope is "uncomfortable," hard to use, gives poor to middlen images, light is hard to get just right, and on... your students will feel the same, and in spades.
We are having trouble to get a good fixation of a putative cop vesicle fraction. Our prim. fixative is 1% glutaraldehyde in 50 mM P-buffer containing 0.05% tannic acid(Sigma T-0125). Postfixation is in 1% osmium in 50 mM P-buffer containing 35% sucrose(from the density gradient). Unfortunately the membranes look fuzzy and it is hard to clearly identify vesicles. Since all fixation protocolls of cop or clathrin coated vesicles seem to contain tannic acid we wonder what we are doing wrong! Any comments are wellcome.
Thank you, Stefan
Dr. Stefan Hillmer Albrecht-von-Haller Institut fuer Pflanzenwissenschaften Universitaet Goettingen Untere Karspuele 2 37073 Goettingen Deutschland
Tel (+49) 551-392013 Fax (+49) 551-397833 e-mail shillme-at-gwdg.de
Bart Cannon wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Hello, } } Does anyone know of a source for bulk volumes of the blue luminescing } plastic scintillator material which is sometimes used in SEM electron } detectors? } } Thanks. } } Bart
Bart, You might want to try Gene Taylor at ME Taylor, tel 1-301-774-6246, fax 1-301-774-6711. He can help with the scintillator material.
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Related to the discussion on toy microscopes, I also have a pet hate in toy binoculars x2 or x3. I know they have to be cheap and plastic, but it irks me that a roughly $15 pair from a "reputable" toy manufacturer performs worse than a $3 set from Hong Kong. This is simply because the "reputable" ones have got a wrong design for the meniscus of the objective.
Next up in price one finds all these 21mm aperature x20 zooms from Tasco, etc. The aperture-magnification ratio is all wrong. One has to go to $200 or so to get a half decent pair. This will put the kids off astronomy, too!
Sorry for intruding a semi-relevant grouch, but on matters like this, I run under OScar in the Grouchputer, the world's most unfriendly computer from Sesame Street Systems:
Garbage In - Yes Please (delete that last word) Gargage Out - Well whad'ya expect!!!
+------------------------------------------------------------------------+ | 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 | +------------------------------------------------------------------------+
As an undergraduate and graduate student in Metallurgical Engineering, I had lots of courses and lectures on SEM, TEM and all the other high-tech microscopy equipment. However, I never had a lecture devoted entirely to light microscopy although we did use it in lots of labs. Although much of the information needed for light microscopy is embedded in the SEM, TEM stuff I think that we all tend to ignore it. If you are teaching undergrads they certainly need it more than the high tech stuff. If you are teaching graduate students they need both the optical and electron microscopy information.
On Sat, 28 Jun 1997 23:37:11 Ron Oleka wrote: } I operate a Philips SEM 505 interfaced to a Tracor Northern 5500 energy } dispersive x-ray spectrometer (EDX). If anyone would like to share their } ideas/methods of interfacing the now almost ancient (but working perfectly) DEC } PDP 11 which is the main guts of the TN5500 I would love to hear from you. Dear Ron, Here at Mektech we are working on adding TN 5500 to the list of analyzers we currently interface to MS Windows platform. We will make the announcement shortly on our website http://www.visionol.net/~mektech (free demo software available there). Mektech Inc. Mektech Inc.
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thanks for the address etc. I already have an excellent microscope from Brunel Microscopes (so if there is anyone connected with them here, you can be assured that I am happy!) I have heard various people say what they think would be an ideal started microscope and low magnifications. I would have thought a monocular with say x20 x50 and x100 would be a good start since x20 is ideal to search with and x100 enlarges the subject to a nice level of detail. I have been doing my first photomicrographs using this magnification.
Considering some 'toy' microscopes can cost up to 50UK pounds I think it would be better for them to junk all the extras in the dearer kits as this seems to be what you pay up to ?30 extra for, as the microscope itself doesn't seem all that different from that sold on its own! Another case of manufacturers enticing kids into thinking 'more' is better! Considering the base model of the microscope that I purchased cost just 79 UKpounds and is solid metal for a start, I am sure that they could at least put glass lenses in the cheap toy ones!
Conrad ------------------------------------------------------------------------ ----------- "Any sufficiently advanced technology is indistinguishable from magic" ----------------------------------------------------------- Arthur C Clarke ----
Tom, Alkaline phosphatase is more sensitive than peroxidase. The reaction product is a dense purple-blue (when using NBT/BCIP). If your signal is quite strong there can be a problem with diffusion of the reaction product, in this case DAB can be a good alternative. See our paper using both NBT/BCIP and peroxidase in J. of Neurosci Methods, 1993, 50:145-152. Linda Barthel Research Associate II Department of Anatomy and Cell Biology University of Michigan lab (313) 764-7476 fax (313) 763-1166 barthel-at-umich.edu
Yes I agree, I think some manufacturers cut TOO many corners. I mean surely a good glass lense shouldn't add that much to the price? But then I guess pennies etc matter more at this level of cost.
I know they always say its best to buy a good quality 'proper' microscope/ telescope etc but for a youngster the price of a good quality piece of equipment may just be a bit too much especially if they do not know if they are going to like it. After all its alright for me to think that 70-100 UK pounds for a microscope is a good price now, but when I was young that was in the realms of professional equipment!
Conrad
} -----Original Message----- } From: Robert H. Olley [SMTP:R.H.Olley-at-reading.ac.uk] } Sent: Monday, June 30, 1997 10:06 AM } To: Microscopy Newsgroup } Subject: OM and Binoculars } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Stefan, In my opinion your problem lies in the field of pure buffer capacity. 50 mM is too low for both tannic acid and glutaraldehyde. Did you check pH?
I would like to suggest the following protocol.
Mix equal volumes of 2% glutaraldehyde in 0.15 M HEPES (pH 7.3) and your fraction. Cetrifudge. Wash with 0.1 M cacodylate buffer (pH 7.3) (I do not know details of your isoaltion procedure). Fix with reduced OsO4. For this mix equal volumes of 2% OsO4 and 3% potassium ferrocyanide in 0.2 M cacodylate buffer (pH 7.3) and then add to your gradient. Thus, solution will contain sucrose from density gradient. Wash with 0.05 M cacodylate buffer (pH 7.0). Treat with freshly prepared 1% tannic acid in 0.05 M CB, wash with 1% NaSO4. Dehydrate and embed.
Sincerely yours, Alexander Mironov fax: +39 872 578 240
On Mon, 30 Jun 1997, S.Hillmer wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear microscopists, } } We are having trouble to get a good fixation of a putative cop vesicle } fraction. Our prim. fixative is 1% glutaraldehyde in 50 mM P-buffer } containing 0.05% tannic acid(Sigma T-0125). Postfixation is in 1% osmium } in 50 mM P-buffer containing 35% sucrose(from the density gradient). } Unfortunately the membranes look fuzzy and it is hard to clearly identify } vesicles. Since all fixation protocolls of cop or clathrin coated } vesicles seem to contain tannic acid we wonder what we are doing wrong! } Any comments are wellcome. } } Thank you, } Stefan } } Dr. Stefan Hillmer } Albrecht-von-Haller Institut fuer Pflanzenwissenschaften } Universitaet Goettingen } Untere Karspuele 2 } 37073 Goettingen } Deutschland } } Tel (+49) 551-392013 } Fax (+49) 551-397833 } e-mail shillme-at-gwdg.de } }
Dutch Society for Microscopy Philips Electron Optics - 5th Cryoworkshop Eindhoven, The Netherlands August 25-29 1997
From 25th to 29th August 1997 we are holding the 5th cryoworkshop at the laboratories of Philips Electron Optics, Eindhoven the Netherlands. The workshop is a blend of practical and theoretical work that has proved very successful in the past.
Practical and theoretical sessions will include :
cryo-ultramicrotomy,immunogold labelling, cryo-electron microscopy, X-ray microanalysis, Electron Energy Loss Spectroscopy (EELS) and Electron Tomography.
Considerable emphasis is placed on the development of practical skills under the guidance of experienced cryo-users using modern TEM's and SEM's with relevant cryo-accessories.Some of the lecture topics include :
"Two dimensional protein crystals in ice" Dr. Alan Brisson, University of Groningen, The Netherlands
"Optimising elemental analysis for life science" Dr. Karl Ziergold, Max-Planck-Institute for Molecular Physiology,Germany
"Three-dimensional image interpretation" Dr. Tim Baker, Purdue University, USA
If you are interested in attending : we will be happy to send you full programme details : please contact Ms. Annemieke Coppelmans by phone on +31 40 2766234 or Fax +31 40 2766102. Alternatively, you can fill in the reply form at our website : http://www.peo.philips.com.
The numbers of participants are limited and all registrations must be recieved by the 31st July 1997.
For general information : please contact Jeremy Rees by phone on +31 40 2766777 or by email at jar-at-eo.ie.philips.nl.
Dr. Jeremy Rees Philips Electron Optics Building AAE, Achtseweg Noord P.O.Box 218, 5600 MD Eindhoven The Netherlands
Caroline has posed a tough problem: a simple and easily available test specimen for microscope optics.
My favorite test specimen is a slide of diatoms of different sizes and frustule patterns. Each species has tiny holes in the frustrules (shells) that are either really small and close together, so visible only at high resolution, or larger and farther apart so visible with poorer optics. These slides and an explanatory key are available in the US from Carolina Biological (800-227-1150, catalog number P7-B25D) but they are expensive, about US $25.75 each. Too expensive and not easily available.
I'm having a tough time thinking of a widely available test specimen that is mostly transparent, but has high contrast, has a gradation of sizes from 0.2 to about 2 micrometers to test resolution, a regular geometric pattern to test other aberrations, and that would be easy for a novice to interpret.
Perhaps a thin onion peel or onion root tip? If you could see vacuoles and nuclei, that might be a good test. Might even see chromosomes.
Gary Radice, Associate Professor gradice-at-richmond.edu Department of Biology 804-289-8107 (voice) University of Richmond VA 23173 804-289-8233 (FAX)
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Three simple steps which can perhaps help.
1. *Before* you go to buy/test a microscope - define what *you* want the microscope to do and in simple task testable terms.
2. If you can talk to/take someone who knows his way around microscopes, do so. Many professional microscopists will be willing to devote some time to "hook" newcomers on something they love.
3. Sit and *play* (no I did not say test, I said play, your students will) with the microscope. Take and examine samples that you plan to use from your decission in step No. 1. *Tune into your impressions*, if the microscope is "uncomfortable," hard to use, gives poor to middlen images, light is hard to get just right, and on... your students will feel the same, and in spades.
Tell me about it. I am trying to talk the Medical School here into letting me give a short course on what to do and how to work that microscope they give to every medical student for their medical school tenure. Seems they feel that there is no room in the student's demand full schedule, AND "everyone knows how to use a microscope."
Go ahead and rant..... SANCHO MY HORSE.
Shalom from Jerusalem, Azriel Gorski
On Mon, 30 Jun 1997, Robin Griffin wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } And one other spin off topic... } } As an undergraduate and graduate student in Metallurgical Engineering, I } had lots of courses and lectures on SEM, TEM and all the other high-tech } microscopy equipment. However, I never had a lecture devoted entirely } to light microscopy although we did use it in lots of labs. Although } much of the information needed for light microscopy is embedded in the } SEM, TEM stuff I think that we all tend to ignore it. If you are } teaching undergrads they certainly need it more than the high tech } stuff. If you are teaching graduate students they need both the optical } and electron microscopy information. } } Just a side rant! }
Following on from the discussion about resolution, I recently had a letter asking whether using diamond (funds permitting) to make all of the optics of a light microscope, in conjunction with a suitable immersion oil might increase the NA and, hence, resolution. I really don't know about the fundamentals of optical microscopy, so thought I might as well throw it out to you all. Anyone bother to comment?
Keith
--- Interface Analysis Centre, University of Bristol, Oldbury House, 121, St. Michael's Hill, Bristol, BS2 8BS, England Telephone: +44 (0)117 925 5666 | Facsimile: +44 (0)117 925 5646 URL: http://www.phy.bris.ac.uk/research/iac/home.html
Dear colleagues, Recently we were asked to analyze the uniformity of hydrogel coating on the surface of the 100 micron particles. The hydrogel consists of 10% bifunctional polyacrylate and some additives. The rest, of course, is water. We froze the samples in liquid N2, and sublimed frozen water using freeze dryer. After SEM observation, teh coatings looked collapsed anyway. This is not what our customer wants. AFM analysis of the obtained samples yields similar results. In addition, it is compounded by the fact that the particles are spherical and the curvature really throws off the small magnificAtion images. We recommended the customer a liquid Tapping Mode AFM (we are not eqiupped with it). Is there any other way we can prepare and analyze the surface morphology of these samples? This is my first time addressing the ListServer, but there is always time for first. Thanks for possible help, Alex Mejiritski Ph. D. Student Center for Photochemical Sciences Bowling Green State University Bowling Green , Ohio 43402 (419) 372-7830
} Date: Mon, 30 Jun 1997 11:24:03 -0500 } To:Richard Thrift {Richard_Thrift-at-depotech.com} } From:ejb11-at-psu.edu (Edward J. Basgall) } Subject:Re: LM: length standard } } } } } Hi } } Can anyone suggest what to use as a length standard with a 100x } } objective? I intend to print out an image of this standard when I print } } images of samples, in order to determine final magnification. I am a little } } concerned with the use of my micrometer slide (10 microns between } } graduations) because the width of the individual lines is significant } } compared to the distance being measured between lines. Also there is } } no tolerance stated for the separation between the lines. I can think of } } using calibrated latex beads suspended in water, but the refractive index } } difference between latex and the aqueous medium (or air) causes a lot of } } problems. Are there other standards I can use? } } } } Thanks } } Richard } } Richard_Thrift-at-Depotech.com } } } Richard, } } I was under the impression that micrometer slides were meant to measure } from one side of a line to the corresponding side on the adjacent line, } not between the lines. Have I been mis-led? } I thought the line thickness is accounted for in this manner. } } } | } | not | { } | } } Cheers } ed } } Edward J. Basgall, PhD } The Pennsylvania State University } Surface Chemistry Group ejb11-at-psu.edu } Materials Research Institute Building Ph: 814-865-0493 } University Park, PA 16802-7003 FAX: 814-863-0618 } }
Does anyone know where I can purchase a Zerostat antistatic pistol? My old source no longer carries them, and I find them really useful in dry sectioning in low humidity (not a problem in Chicago in the summer, but in the winter...) and in keeping grids from bouncing onto the lid of the Petrie dish. Cheers from Joyce Craig Chicago State University
Hi Larry, Regarding colour variation with silicon crystal orientation; silicon is isotropic, hence the colours of silicon in transmission are identical for any crystal orientation. We have a very pretty picture of this (lack of) effect in a more thorough investigation of "thickness fringes and the colours of silicon in transmission" which has just been submitted. ("More thorough" relative to a short note now out - MICRON, Vol. 27, No. 6, pp. 407- 411 (1996)). A more likely source of differences in quoted transmission colours for silicon is the illumination source. the colour scheme looks something like this:
A = CIE standard illuminant A; gas-filled tungsten lamp, color temp. of 2854K. C = CIE standard illuminant C; average daylight, with a color temp. of 6770K.
Illumination source -} A C Thickness (microns)
0 yellow clear
1 light orange orange/yellow
2 orange light orange
3 orange/red orange
4 reddish orange orange/red
5 red reddish orange
more deeper and darker red deeper and darker red
Notice that the colours for light source A "lag" the colours for light source C by about 1 micron; i.e., the transmission colour for silicon at 1 micron for light source A is nearly identical to the transmission colour for light source C at 2 microns.
Cheers John ----------------------------------------------------------------------- - | John P. McCaffrey tel: Canada 613-993-7823 | | National Research Council of Canada fax: Canada 613-990-0202 | | Inst. for Microstructural Sciences _____ _____ | | Montreal Road Labs, Bldg. M-50 | | __/\__ | | | | Ottawa, Ontario, K1A 0R6 | | __/\\ //\__ | | | | Canada | | \ \\ // / | | | | | | /___ ___\ | | | | email: john.mccaffrey-at-nrc.ca | | /__ __\ | | | | | | || | | | | |_____| |_____| | ----------------------------------------------------------------------- - ------------------------------------------------------------------------ ------ REPLY FROM: McCaffrey, John (IMS) ----------------------------------------------------------------------- - The Microscopy ListServer -- Sponsor: The Microscopy Society of America ----------------------------------------------------------------------- .
} Dear microscopists, } Does anyone know of a reliable study relating color to the thickness of } Silicon ( {20 micron)? I have been using one set of values, and then came } across another set which was quite different, and I am now wondering which } (if either) is correct. Thanks in advance for your time and help. } } Mick Thomas } Materials Science Center } Cornell University
Don't know the answer to your problem but ....
My understanding is that the colour at various thicknesses is dependent on, among other things, the bandgap, which in turn is orientation dependent. So, 100 Si will have a different colour/thickness dependence compared with 111 or 110.
This might account for the different valuses you have seen.
Hi Larry, Regarding colour variation with silicon crystal orientation; silicon is isotropic, hence the colours of silicon in transmission are identical for any crystal orientation. We have a very pretty picture of this (lack of) effect in a more thorough investigation of "thickness fringes and the colours of silicon in transmission" which has just been submitted. ("More thorough" relative to a short note now out - MICRON, Vol. 27, No. 6, pp. 407- 411 (1996)). A more likely source of differences in quoted transmission colours for silicon is the illumination source. the colour scheme looks something like this:
A = CIE standard illuminant A; gas-filled tungsten lamp, color temp. of 2854K. C = CIE standard illuminant C; average daylight, with a color temp. of 6770K.
Illumination source -} A C Thickness (microns)
0 yellow clear
1 light orange orange/yellow
2 orange light orange
3 orange/red orange
4 reddish orange orange/red
5 red reddish orange
more deeper and darker red deeper and darker red
Notice that the colours for light source A "lag" the colours for light source C by about 1 micron; i.e., the transmission colour for silicon at 1 micron for light source A is nearly identical to the transmission colour for light source C at 2 microns.
Cheers John ----------------------------------------------------------------------- - | John P. McCaffrey tel: Canada 613-993-7823 | | National Research Council of Canada fax: Canada 613-990-0202 | | Inst. for Microstructural Sciences _____ _____ | | Montreal Road Labs, Bldg. M-50 | | __/\__ | | | | Ottawa, Ontario, K1A 0R6 | | __/\\ //\__ | | | | Canada | | \ \\ // / | | | | | | /___ ___\ | | | | email: john.mccaffrey-at-nrc.ca | | /__ __\ | | | | | | || | | | | |_____| |_____| | ----------------------------------------------------------------------- - ------------------------------------------------------------------------ ------ REPLY FROM: McCaffrey, John (IMS) ----------------------------------------------------------------------- - The Microscopy ListServer -- Sponsor: The Microscopy Society of America ----------------------------------------------------------------------- .
} Dear microscopists, } Does anyone know of a reliable study relating color to the thickness of } Silicon ( {20 micron)? I have been using one set of values, and then came } across another set which was quite different, and I am now wondering which } (if either) is correct. Thanks in advance for your time and help. } } Mick Thomas } Materials Science Center } Cornell University
Don't know the answer to your problem but ....
My understanding is that the colour at various thicknesses is dependent on, among other things, the bandgap, which in turn is orientation dependent. So, 100 Si will have a different colour/thickness dependence compared with 111 or 110.
This might account for the different valuses you have seen.
Mick, there was a homepage, I believe at Rice University of someone who had a picture with a thickness bar below it. Unfortunately I lost the link.
I believe they had wedged a sample at a known angle and calculated the thickness that way. I have done some work on dimpled samples in the past. My calculations have always been that for p- silicon amber is at about 12 microns going almost linearly to white at {1 micron.
Good luck
Lucio
Dr.Lucio Mule'Stagno MEMC Electronic Materials Inc University of Missouri -St.Louis Silicon Materials Research Group Physics Dept., 501 Pearl Dr., 8001 Natural Bridge rd., St.Peters, St.Louis MO 63376 MO 63121 tel: 314 279 5338 314 516 5931/3 fax: 314 279 5363 314 516 6152 email: lmulestagno-at-memc.com luciom-at-newton.umsl.edu
My suggestion would be to find someone with a cryo stage on their SEM and look at the sample in the frozen, fully hydrated state. } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } At 10:50 AM 6/30/97 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
Reinhard, Thank you for the correction. I was referencing "Low Temperature Methods in Biological EM" A.W. Robards and U.B. Sleytr in Practical Methods in Electron Microscopy, A.M. Glauert, ed.1985.
Dubuchet and McDowell 1981 are cited as working with pure water to determine the low recystallization temp of 130K (p11) and "The temperature below which pure ice does not recrysatallize is as low as 130K but this is temperature-dependent and recrystallisation probably does not take place at significant rates until the temperature is above 180-190K "(Moor, 1973; Nei 1973). (p19)
Later, on page 253, the book states: "As discussed in Chapter 2, for most biological specimens with an average water content, recrystallization phenomena can be expected at temperatures above -80C (193K), but since recrystallization is also a time-dependent phenomenan, no detectable damage may occur during short drying periods at higher temperatures."
I'll have to look at the more recent references...
Ed Basgall
} } I would opt for the freeze dryer that can hold the lowest temp. If you are } } using aqueous suspensions you want to remain below the ice recrystalization } } point ( { -80C). } } Just to remind everybody that the recrystallization of vitrified water } has been directly observed to take place at a temp of at least } 135 - 140 K, i.e. -138 C to -133 C. } Lit: Dubochet and MacDowell 1981 } Dubochet and Lepault 1984 } see also: Bachmann and Mayer, 1987, Chapter 1, in: Cryotechniques in Biological } EM (Steinbrecht and Zierold, ed) Springer Berlin } } Dr. Reinhard Rachel {Reinhard.Rachel-at-biologie.uni-regensburg.de} } Universitaet Regensburg, Lehrstuhl fuer Mikrobiologie } D - 93040 Regensburg (Tel.: xx49-941-943-4534) } http://www.biologie.uni-regensburg.de/Mikrobio/Stetter } http://www.biologie.uni-regensburg.de/Mikrobio/Stetter/Gruppen/rachel.html
} Following on from the discussion about resolution, I recently had a letter } asking whether using diamond (funds permitting) to make all of the optics } of a light microscope, in conjunction with a suitable immersion oil might } increase the NA and, hence, resolution. I really don't know about the fun- } damentals of optical microscopy, so thought I might as well throw it out } to you all. Anyone bother to comment? } Dear Keith, Since the refractive index is 2.4, this could give a high NA. Whe- ther there is a suitable oil or whether there is anything to gain by making lenses other than the objective out of diamond, I do not know. Another pos- sible difficulty could be chromatic aberration--I don't know the dispersion for diamond or whether that would be easy to correct. It goes without say- ing that there would be some minor fabrication problems :-). Yours, Bill Tivol
When I started my position here I inherited a Microanatomy course. At the time it was kind of an antique--basically a medical school type survey of mammalian cell and tissue types. Not usually taught to undergraduates anymore. The microscopes were crummy so the previous instructor taught the whole thing using Kodachrome slides.
With the help of NSF, I was able to get new microscopes and a couple of Macintosh imaging workstations with videocameras. I have students fix their own tissues, embed, section, and stain their own preps. Everyone also does SEM preps, and a few students can do TEM as an option. They all learn how to clean and align a microscope, and they all learn K=F6hler illumination. I= n fact, I test them three times on K=F6hler illumination so they all know it cold by the end of the course. I go over enough optics so that they most of them understand things like the difference between refraction and diffraction, what a focal point is, where the real and virtual images are formed, different lens aberrations, and conjugate image planes. (Some are surprised to see that their physics course is actually relevant to biology.) I cut back on the tissue survey aspect of the course and emphasize using microscopy as tool to understand tissue function.
I thought that it wouldn't be a very popular course since it is not very molecular, but I can tell you that students LOVE it. They like learning something practical for a change, they take great pride in their specimens, they like working with something they can see. Those that have gone on to medical or vet school report that they are WAY ahead of other students when it comes to histology class.
Anyway, I know I am preaching to the choir here but I definitely believe that teaching light microscopy has an important place in a basic biology curriculum, because it remains a basic tool. If you need an argument for your administrators, tell them that knowing how to use a microscope gives your students an edge over their competitors.
Gary Radice, Associate Professor gradice-at-richmond.edu Department of Biology 804-289-8107 (voice) University of Richmond VA 23173 804-289-8233 (FAX)
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} Azriel -
It's more like "everyone knows how to MISUSE a microscope, isn't it? The only classroom technology that gets respect these days is the computer, so if you can't lick 'em, join 'em. There's a new CD-ROM (Windows & Mac) out from a reputable source (Center for Bioengineering, U. of Washington, Seattle) that may do some good (tho I haven't had a chance to review it yet for the Project MICRO bibliography). Microscopy-Tutor, Lippincott-Raven, ISBN 0-7817-1217-3, $195.00. http://www/lrpub.com, 800-777-2295.
Caroline
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
Keith wrote from Bristol: } Dear All, } } Following on from the discussion about resolution, I recently had a letter asking } whether using diamond (funds permitting) to make all of the optics of a light } microscope, in conjunction with a suitable immersion oil might increase the NA } and, hence, resolution. I really don't know about the fundamentals of optical } microscopy, so thought I might as well throw it out to you all. Anyone bother to } comment? } } Keith
---
Being the token optical engineer on this list makes me the token lens designer, too :) Although I have never designed an immersion type microscope objective the general trend is that as you go up in index of refraction the job gets easier. A high index "glass" that won't etch should make the lens design practical. Of course the high index of diamond should make it possible to get higher NA's
I remember seeing an advertisement recently about a stereoscope set up for fluorescence microscopy. Does anyone have experience with such a configuration? Any recommendations? In our lab we are very often limited not by how high we can go in magnification but how low. Another good zoom stereoscope might help a lot, especially if we could image fluorescent objects. Most of our immediate applications would require UV excitation; however the typical FITC & rhodamine bands would also be useful. But of course I don't want to spend much money :-) An alternative might be some sort of UV lighting setup for my video camera and macro lens. Any recommendations on this option?
Any tips would be appreciated! And thanks to all who responded to my questions about glass strips and video printer problems!
Karen
-- Karen Zaruba kszaruba-at-mmm.com 3M Company, 3M Center Bldg. 270-1S-01 St. Paul, MN 55144 "The opinions stated above are my own, not necessarily 3M's"
On a related subject, has anyone else noticed that Si samples get darker when you start to ion mill them? I usually get TEM cross-sections down to ~8-10 um, and when the sample goes in the mill it's an amber/red colour (from the rather poor light shining through it). Ten minutes later it's almost opaque. Maybe it's just the surface roughness increasing; or perhaps the effect of an amorphous layer? And while I'm on the subject of looking through samples in the ion mill, has anyone come up with a way of seeing how thick GaAs or InP are while milling? GaAs seems to become transparent at about 1 um (ish), and InP always looks dark no matter what. A real pain when setting the timer!
Cheers,
Richard Beanland GMMT Ltd., Caswell, Towcester, Northants NN12 8EQ UK
} Recently we were asked to analyze the uniformity of hydrogel coating on the } surface of the 100 micron particles. The hydrogel consists of 10% bifunctional } polyacrylate and some additives. The rest, of course, is water. We froze the } samples in liquid N2, and sublimed frozen water using freeze dryer. After SEM } observation, teh coatings looked collapsed anyway. This is not what our } customer wants. AFM analysis of the obtained samples yields similar } results. In } addition, it is compounded by the fact that the particles are spherical } and the } curvature really throws off the small magnificAtion images. We recommended the } customer a liquid Tapping Mode AFM (we are not eqiupped with it). Is there any } other way we can prepare and analyze the surface morphology of these samples? } This is my first time addressing the ListServer, but there is always time for } first. } Thanks for possible help, } Alex Mejiritski } Ph. D. Student } Center for Photochemical Sciences } Bowling Green State University } Bowling Green , Ohio 43402 } (419) 372-7830 }
How about an Environmental SEM? You could look at the sample in a fully hydrated state, then slowly dehydrate, to reveal sub-surface structure.
Ed is quite right, one space plus one line. If possible, the accuracy is improved if many spaces/lines and are included. A stage micrometer could be used to establish the magnification and for a given lens combination that figure would be "permanent". For some uses it could be most useful to then establish the length of the negative in micrometers and use that as a standard, but some people would prefer large latex spheres incorporated with the specimens. For low power SEM and reflected LM there is a 0.01mm graduated scale available for calibration of those instruments (See Pelco or ProSciTech). Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 350+ Links, MSDS ************************ http://www.proscitech.com.au } } } Can anyone suggest what to use as a length standard with a 100x } } } objective? I intend to print out an image of this standard when I print } } } images of samples, in order to determine final magnification. I am a little } } } concerned with the use of my micrometer slide (10 microns between } } } graduations) because the width of the individual lines is significant } } } compared to the distance being measured between lines. Also there is } } } no tolerance stated for the separation between the lines. I can think of } } } using calibrated latex beads suspended in water, but the refractive index } } } difference between latex and the aqueous medium (or air) causes a lot of } } } problems. Are there other standards I can use? } } } } } } Thanks } } } Richard } } } Richard_Thrift-at-Depotech.com } } } } } } Richard, } } } } I was under the impression that micrometer slides were meant to measure } } from one side of a line to the corresponding side on the adjacent line, } } not between the lines. Have I been mis-led? } } I thought the line thickness is accounted for in this manner. } } } } } | } | not | { } | } } } } Cheers } } ed } } } } Edward J. Basgall, PhD } } The Pennsylvania State University } } Surface Chemistry Group ejb11-at-psu.edu } } Materials Research Institute Building Ph: 814-865-0493 } } University Park, PA 16802-7003 FAX: 814-863-0618 } } } } } }
What about a piece of black and and white photographic negative film for a cheap test specimen? On a good microscope, the silver grains appear with sharp edges. I don't have a toy microscope around to compare. But film is widely available and cheap, dry, and you don't even need a microscope slide or coverslip. No regular pattern to detect barrel or pincushion distortion, but these probably wouldn't matter to most users anyway.
BTW, someone around here made up a bunch of cheap stage micrometers by photographing a metric ruler from a distance with a 35 mm camera. With the reduction, the one mm divisions on the scale are 100 micrometers apart on the negative. These were then just cut up and glued to microscope slides. They are a little fuzzy but if you measure center to center of each line it is close enough. Perhaps one could photograph a grid to make a test specimen for distortion.
Gary Radice, Associate Professor gradice-at-richmond.edu Department of Biology 804-289-8107 (voice) University of Richmond VA 23173 804-289-8233 (FAX)
David - use the find function on your browser and search for "Protist" on our link page. That link takes you a sites at the Uni of Montreol which has the very things you are looking for. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 350+ Links, MSDS ************************ http://www.proscitech.com.au
---------- } From: David Webb {davehawaiiedu-at-msn.com} } To: Microscopy-at-sparc5.microscopy.com } Subject: Images of Algae and Sea Weeds } Date: Monday, 30 June 1997 5:12 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I am looking for all types of images (light & EM) which deal with algae, } especially sea weeds. I plan to teach a course in which I will compare and } contrast vegetative and reproductive adaptations of algae with land plants. } The emphasis will be at the light microscope level, but SEM and TEM images } which deal with major aspects of form and function would be welcome.
Cleaning grids? Go your hardest if you have nothing better to do: Collect grids in a 20 or 30ml glass vial. Soak them in chloroform, give one change in chloroform, pour off solvent, replace with ethanol or acetone, replace with water, add drops of detergent, ultra sonicate for a short time. Rinse with water, etch for a moment in weak acid (10% acetic or 0.2N HCl) rinse in distilled water. Pour water off. Use washbottle with water (or for faster drying ethanol) to flush grids into a Petrie dish lined with filterpaper; pipette off excess fluid. Sit dish in an incubator until dry. Thousands of grids can be treated in one batch but the joy is in sorting the grids and throwing the bad once out. We sell standard copper grids at A$10/vial/100; which is a little over US$7. In a labour-costly country, with the odd exception, cleaning grids is a waste of time. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 400+ Links, MSDS ************************ http://www.proscitech.com.au
} I don't know about the first, I've never had really good results from } cleaning off coated grids, but as to the second; in the absence of a Glow } Discharge apparatus you might get some result out of using a Zerostat } Antistatic Pistol. It is not so reliable as Glow Discharge and takes some } practice to obtain neutral charge grids. } } They are still available as far as I know. We bought one recently for } zapping our Balance after frustrating sessions of attempting to weigh out } powders which were flying all over the bench. It cured that. } } If you want to see if it works before buying, find yourself a Black Vinyl } Record collector and borrow one. They often use them for discharging static } on Discs before playing. I used one myself before going over to CD's many } years ago. } Regards } Stephen Griffiths } } {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} } Stephen Griffiths e-mail:- s.griffiths-at-ucl.ac.uk } Visual Science Department Phone:- 0171 608 6914 } Institute of Ophthalmology Fax:- 0171 608 6850 } Bath Street, London. EC1V 9EL } {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} } } } Just wondering about a couple of things. } } First the different ways there are to clean grids that have a } } film (2% parlodion), and a carbon coat, but no sample. } } Second the best way to reduce or eliminate static on coated grids, } } without using a glow discharge apparatus. } } Thanks, } } Maya Moody }
We still have a "bunch" available. See our on-line catalogue. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 400+ Links, MSDS ************************ http://www.proscitech.com.au ---------- } From: joyce craig {bafpjec-at-csu.edu} } To: Listserver {Microscopy-at-sparc5.microscopy.com} } Subject: Cleaning grids (Zerostat) } Date: Tuesday, 1 July 1997 15:18 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Does anyone know where I can purchase a Zerostat antistatic pistol? My } old source no longer carries them, and I find them really useful in dry } sectioning in low humidity (not a problem in Chicago in the summer, but } in the winter...) and in keeping grids from bouncing onto the lid of the } Petrie dish. } Cheers from } Joyce Craig } Chicago State University
From Mark Lund: } } --- } } Being the token optical engineer on this list makes me the token lens } designer, too :) Although I have never designed an immersion type } microscope objective the general trend is that as you go up in index } of refraction the job gets easier. A high index "glass" that won't } etch should make the lens design practical. Of course the high index } of diamond should make it possible to get higher NA's
But diamond has a high dispersion relative to quartz (0.044 vs 0.013). Wouldn't this make correcting for chromatic aberration difficult?
How about corundum? RI = 1.762 -1.77, dispersion = 0.018
On a separate note: as the "token optical engineer", have you been following the thread about microscopes for kids and lower-cost-but-still-quality ones for amateurs?
Phil
} Sic Hoc Legere Scis Nimium Eruditionis Habes { Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217) 355-1143 oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
The index of refraction of diamond is 2.4173. I don't know as much about the theory of microscope optics as I should but I do know that this plays a role.
Joe Tabeling Delaware Diamond Knives 3825 Lancaster Pike Wilmington, DE 19805 302-999-7476
The server was down for nearly a full day while I replaced the main drive. Expect a hic-cup or two while things settle in.
The subscribe/unsubscribe functions should (?) be functioning tomorrow AM. For those of you that have tried to unsubscribe you should be "off" the list tomorrow.
I apologize for the inconvenience...
Nestor Your Friendly and Blurry Eyed Neighborhood SysOp
We want to do immunohistochemistry on human heart muscle tissue cryosections. Which embedding medium should be used for cryosectioning on a Leica cm 3000 kryostat for light microscopy? Are there simple recipies and procedures (cryoprotection...)? Which freezing techniques should be used? We get the tissue in the operating theatre which freezing technique can be used directly there? I sthere a freezing technique for both, cryosectioning and RNA-preparation of the same tissue-block.
Thank you all for your answers
Christoph Guenther Klinikum der Charite der Humboldt Universitaet Ziegelstrasse 5-7 10117 Berlin /Germany Tel.: +49 30 2802 6327 Fax + 49 30 2802 6608
} There's a new CD-ROM (Windows & Mac) out } from a reputable source (Center for Bioengineering, U. of Washington, } Seattle) that may do some good (tho I haven't had a chance to review it yet } for the Project MICRO bibliography). Microscopy-Tutor, Lippincott-Raven, } ISBN 0-7817-1217-3, $195.00. http://www/lrpub.com, 800-777-2295.
The URL for Microscopy-Tutor is "http://www.lrpub.com/media/m1208.htm".
Here's an advertising blurb from the web site.
============================================== Microscopy-Tutor, CD-ROM for PC and Macintosh, with Two-Color Insert, by The Department of Laboratory Medicine and the Center for Bioengineering, University of Washington, Seattle, WA.
This interactive computer program guides students of biology and the health professions through the basic concepts of bright field light microscopy, developing and refining the user's knowledge by providing a more active role in learning. Simple, approachable, and largely qualitative, Microscopy-Tutor uses three-dimensional animation to simulate a microscope with an integrated illuminator and adjustable field diaphragm. The CD-ROM's animated diagrams are more accurate than those found in most university-level texts, but are also easy-to-understand because key concepts and equations are presented visually rather than symbolically. Since using Microscopy-Tutor feels more like operating a microscope than a computer it's successful in presenting changes in dynamic processes that occur during alignment and use of the microscope. Rather than explaining what happens through the use of words and pictures, the user -- who learns by doing -- can see what happens and better understand the implications. In some sections two animated perspectives are synchronized, allowing the viewer to change microscope settings and simultaneously see the resulting image. Interactive quizzes test the user's knowledge and accent areas for improvement. ==============================================
The information below is slightly wrong. Not a big deal, but in the interest of science I thought I would set things slightly straighter :)
Azriel Gorski wrote tht McCrone and McCrone wrote:
} Lenses have several types of aberrations which will cause the loss of } detail unless corrected for. Toy microscopes are not. } } Spherical aberration - "is especially apparent in lenses having sperical } surfaces. Light paths near the center of the lens focus at different } points compared to light paths near the periphery."
The name "spherical aberration" has historical roots in astronomy where a spherical mirror has this aberration but a parabolic mirror does not. In some instances a spherical mirror will have no spherical aberration wheras a parabolic mirror will have maximum spherical aberration. The aberration really has nothing to do with the spherical surfaces of lenses. The confusion comes because making a surface "aspheric" can cure spherical aberration in many instances.
} Field curvature - "is a natural result of using lenses with curved } surfaces. The image plane produced by such lenses will be curved. This } kind of image occurs in microscopy unless plano (flat-field) objectives } are used."
Actually, field curvature is a natural result of the geometry of the real world. Since an object at the edge of the field of view is farther from the lens "center" it will tend to be focussed closer to the lens than an object on axis. This naturally leads to field curvature unless the designer makes the lens weaker for off axis points. It has nothing to do with the lenses being curved. The confusion comes from the formula for Petzval curvature, which has lens powers in it.
To the amateurs - welcome! We need you to keep us fresh and excited in what we do.
The alk phos is more sensitive because it will continue to reduce substrate long after peroxidase has quit.
Bob Morphology Core
On Fri, 27 Jun 1997, Tom Phillips wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } We are starting some in situ work with digoxigenin labeled probes. A lot } of the kits and studies seem to use Alkaline Phosphatase labeled antibodies } to detect the digoxigenin. I am curious why they seem to prefer Alk Phos } over peroxidase coupled antibodies which are the more standard } immunocytochemical choice. Anybody have any thoughts on this? } } } Thomas E. Phillips, Ph.D. } Associate Professor of Biological Sciences } Director, Molecular Cytology Core Facility } 3 Tucker Hall } University of Missouri } Columbia, MO 65211 } (573)-882-4712 (voice) } (573)-882-0123 (fax) } } }
Talk about a step backwards... Historically, some objectives were made with different gems in the 1800's (I believe, I don't have my history references at work), but they suffered from several problems. First there are problems with the optical clarity of the gem, color in the case of rubys and garnets, polishing the surfaces to the optimum shape and of course, cost. In the case of polarized light, the double refractive index stones were disastrous. This development did not lasted very long as it was too impractical. Also remember, an immersion system requires more than oiling the front lens of the objective. The condenser should be oiled to the slide as well as the objective. In these systems the resolution is limited by the lowest refractive index, which for a diamond condenser and objective could be the immersion oil.
I think Aldrich still sell them, catalogue number: Z10,881-2, I've only got an old catalogue, and Aldrich might be your old source, so this might not be too useful.
Ray
At 10:18 pm -0700 30/6/97, joyce craig wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Ray Hicks ________________________________________________________________________ |University of Cambridge |Tel 01223 330149 | |Department of Medicine |Fax 01223 336846 | |Level 5, Addenbrookes Hospital |e-mail {rh208-at-cus.cam.ac.uk} | |Hills Road Cambridge |Web http://facsmac.med.cam.ac.uk | |CB2 |ftp server ftp://131.111.80.78 | |UK | | |_________________________________|_____________________________________|
I have looked at the discussion on recrystallization with interest. As I am an agreeable fellow, I would say that you are both correct!
It is true that amorphously vitrified pure water will devtrify at about -130 C or lower, depending partly on the rates of cooling and rewarming. The other extreme was described by MacKemzie AP (1981) Modellling the ultra-rapid freezing of cells and tissues. In: Microprobe analysis of Biological Specimens (eds) Hutchinson TE & Somlyo AP. Academic Press, NewYork, London, 397-421. In this work, which used starches and gels, some specimens could be rewarmed to about -6 C before crystal growth took place (i.e. in the higher molecular weight substances).
I published one micrograph in Scanning Microsc. 6, 715-743 (KP Ryan, Cryofixation of tissues for electron microscopy: a review....) in 1992 which showed some fish spleen tissue elements stored at -60 for 48 hopurs which showed no observable signs of crystal growth. Other results (some published only in my thesis) showed that the samwe tissue could be stored at -40 for some days but after 8 days there were signs of crystal growth in the extracellular fluid (more 'watery'?).
This is an interesting aspect of 'cryo' and deserves more attention by someone. I only did a piece of this to validate the 48 h at -80 C freeze substitution that I used. It should not induce crystal growth. The same f/s has been done at -50 C without apparent crystal development (but not by myself). In tthat case I presume the substitution got to the frozebnwater before it had time to show growth in the crystals
Dear colleagues: All of our scopes are JEOL made and we have been using filaments made by JEOL. The engineers from JEOL also recommend us to use their parts. As far as I know, however, there are companies which also sell filaments. Could anyone of you give me some information as to which source is better than the others? Thank you very much. Regards, Yuhui
} } From Mark Lund: } } } } --- } } } } Being the token optical engineer on this list makes me the token lens } } designer, too :) Although I have never designed an immersion type } } microscope objective the general trend is that as you go up in index } } of refraction the job gets easier. A high index "glass" that won't } } etch should make the lens design practical. Of course the high index } } of diamond should make it possible to get higher NA's } .But diamond has a high dispersion relative to quartz (0.044 vs 0.013). } Wouldn't this make correcting for chromatic aberration difficult? } } How about corundum? RI = 1.762 -1.77, dispersion = 0.018 } } On a separate note: as the "token optical engineer", have you been } following the thread about microscopes for kids and } lower-cost-but-still-quality ones for amateurs?
First: sorry about my first post. My computer decided that it had more important things to think about and while I was trying to get its attention the message posted before I could finish it.
The important thing is not the dispersion--many glasses have higher dispersion than diamond, but that the dispersion is much higher than you would expect for a refractive index of 2.4. Not only is the dispersion anomalous, but the partial dispersions are all well off the normal "glass line." This means that making a lens where one of the elements is diamond would be much easier than otherwise. Not only would correcting the color be easier but correcting residual color (i.e. making the lens apochromatic) would be easier. Also, for an immersion lens (with the right index matching oil) you could probably get an NA greater than 2.0, maybe even as high as 2.3. Of course the condensor and slide would also need to be high index for this to be useful :(
It looks like a fun idea, let me know if you need someone to design one!
Sapphire would be the perfect first element except for its natural birefringence--it would come out astigmatic on axis.
best regards mark
Mark W. Lund, PhD Director } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"The state is good at simple tasks, like killing people and seizing their wealth. It has far more trouble reaching inside individuals and making them good." Doug Bandow
With regard to cutting undecalcified bone samples (with and without implants), a lot of people have found the best method of achieving high straightness, low damage, thin cuts is with an annular diamond blade.
This blade involved in this process is held in a precision chuck and mounted at high tension, to give very little lateral movement, thus producing the high quality of cut. There are many exponents of the method in the hard tissue & dental research fields.
I'd be pleased to send you a relevant technical article on the use of this system for this application and also further information on the commercial version of the annular cutting machine, the 'Microslice 2'. Please send me your address details so I may get the information through to you.
(Note to Group: This product is sold by us -- Please accept this as a notification of our commercial interest in this product)
I look forward to hearing from you.
Best Regards
Tim Hazeldine Product Manager, ULTRA TEC Mfg., Inc. ...............................................................the cutting and polishing specialists 1025 E. Chestnut Avenue Santa Ana CA 92701 - 6425 USA Tel. +1 714 542 0608 Fax. +1 714 542 0627 e-mail. info-at-ultratecusa.com or ultratecex-at-aol.com .............................................................................. .....................................
} The dispersion is 0.044. } In units of per nanometer? In any case, I still don't know what effect this might have on the practicality of using diamond lenses. If the illumination were single-wavelength, there would be no problem.
} Fabrication problems? } Yes, but a colleague suggests that one can use diamond powder as a grinding agent. You at DDK would know a lot more than I. On the plus side, diamond lenses would be very hard to scratch. Yours, Bill Tivol
Recently we did an SEM/EDX analysis that gave rather baffling results. After some thought we came up with what seemed to be a plausible explanation, however, the people for whom the analysis was done remained skeptical. Therefore I am writing the Listserver for a second opinion. The sample consisted of irregular shaped alumino-silicate particles roughly 2-5 microns is size. The specimen was uncoated resulting in some charging. Analysis (windowless) at 1000x mag. resulted in a carbon peak much larger than the Al or Si peaks. At 5000x the carbon peak was smaller; i.e., comparable to the Al and Si. Using a spot mode on individual particles, no carbon was detected. However, spot mode analysis after lowering the accelerating voltage from 20 to 10 kv, showed a small carbon peak. The only explanation we could envision to fit these results was a thin carbonaceous coating on the particle surface. Is this a plausible explanation or we missing something? Does anyone have an alternative suggestion? Any help would be appreciated.
Dan Schwab Center for Microscopy and Microanalysis Bowling Green State University Bowling Green, OH
Leica (and probably other manufacturers) sells NIST (US) or NPL (UK) traceable stage micrometers, certified to tenths of a micron. Measurements are from centers of the lines. They're not cheap.
If you want to do your own certification, start with some regular fine grid, perhaps a dime store holograph, measure it against some standard you trust, and figure out the spacing.
I have just today found a company that supplies the Zerostat antistatic pistol in the U.S.
Bellex International Voice: (302) 761-9885 FAX: (302) 761-9896
The company is located in Wilmington, DE. Ask for Deborah Hunt.
-=W.L. Steffens=- Department of Veterinary Pathology College of Veterinary Medicine University of Georgia Athens, GA 30602 USA http://www.vet.uga.edu/vpp/wls/steffens.html
There are at least two problems involved in storing objects (specimens, pole pieces, specimen holders, and other things commonly stored in dessicators) in a plastic vacuum dessicator under vacuum: (1) the dessicator may have a small leak, and over time come back up to atmospheric pressure, thus exposing the objects to ordinary air, (2) under the influence of the vacuum plasticizers may bleed out of the plastic from which the desiccator is made and find their way onto the stored objects, as noted by others.
Both of these problems can be minimized quite simply, however, by filling the dessicator to atmospheric pressure with a dry, inert gas, rather than leaving it evacuated. For most purposes dry nitrogen would be a satisfactory gas to use here, although helium or argon might be preferred for storing some reactive materials. If the gas is purchased in a high pressure tank it is necessary to be sure that it is oil-free, otherwise the fill-gas may carry oil vapors onto the stuff you are storing inside the dessicator, thereby defeating the purpose of the whole operation, a matter discussed on p. 64 of my book 'Vacuum Methods in Electron Microscopy'.
The vapor pressure of water at the temperature of liquid nitrogen is in the realm of 10-20 Torr, and the vapor pressure of most oils is even lower, and so the gas that is constantly boiling off each container of liquid nitrogen is about as clean and dry as any you can get. As described on p. 65 of Vac Meth in EM, this dry nitrogen can be collected and used to fill vacuum apparatus rather simply. Fit a one-hole stopper into the LN2 storage flask and connect it to the inlet valve of the vacuum container with non-collapsable, flexible tubing (ordinary polyethylene tubing works well). Attach a large, collapsable plastic beach ball to a Tee joint in this tubing with a length of soft, surgical-rubber tubing, and make a clean slit in this surgical tubing about 100 mm long with a sharp razor blade or scalpel. Ordinarily this slit will close thghtly enough so that the nitrogen gas evolved from the storage container will be directed into the beach ball; however, if the ball becomes full the slit will serve as a primative pressure-release valve by opening slightly and allowing the gas to escape so that there is no danger of over-pressurization. When the inlet valve to the evacuated chamber is opened, the nitrogen will flow into the chamber only under the influence of atmospheric pressure acting on the collapsable beach ball, and so there is no danger of exceeding atmospheric pressure in the chamber. An ordinary beach ball will hold enough gas to fill most dessicators several times.
If an oil-sealed rotary vane pump is used to evacuate the storage chamber then some care must be exercised to avoid having oil vapours backstream from the pump into the chamber. To avoid this, it is important not to pump the chamber down below the range of viscous flow (i.e. below about 0.1 Torr - see p.29 of Vac Meth in EM). If this is not considered to be a sufficient vacuum to remove as much atmospheric gas as desired, then the container can be filled with the dry gas, pumped out and filled again a couple of times. Each time the container is pumped out some 90% of the existing gas molecules are removed, and so two or three flushings should leave only an insignificant trace of the original atmospheric gases.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
On Jul 1, 4:09pm, Christoph Guenther wrote: } We want to do immunohistochemistry on human heart muscle tissue cryosections. } Which embedding medium should be used for cryosectioning on a Leica cm } 3000 kryostat for light microscopy? } Are there simple recipies and procedures (cryoprotection...)? } Which freezing techniques should be used? We get the tissue in the } operating theatre which freezing technique can be used directly there? I } sthere a freezing technique for both, cryosectioning and RNA-preparation of } the same tissue-block.
In skeletal muscle, we embed in OCT compound (Miles, Inc.) and plunge freeze in melting isopentane. To avoid freeze artifact, go with smaller pieces of tissue ( { 20-30 mg). Also, keep the specimen in the melting isopentane only for 8-12 sec otherwise sample cracking occurs. For more detail, see the "Color Atlas of Muscle Histochemistry" by R.A. Brumback and R.W. Leech (PSG Publishing Co., 1984).
-- Gordon L. Warren, Ph.D. Research Scientist Muscle Biology Laboratory 158 Read Building Texas A&M University College Station, TX 77843-4243 Fax (409) 862-4808 Phone (409) 862-4809 office e-mail root-at-rangers.tamu.edu
He remarked that freeze-drying (for SEM) produced collapsed coatings and that AFM (imaging in air) produced the same result.
} We recommended the } customer a liquid Tapping Mode AFM (we are not eqiupped with it). -- I am happy to say that ASM is equipped to do exactly this. We have had good success imaging wet samples.
Don Chernoff Analytical Services Division
Advanced Surface Microscopy, Inc. E-Mail: asm-at-indy.net 6009 KNYGHTON RD. Voice: 317-251-1364 INDIANAPOLIS IN 46220 Toll free: 800-374-8557 (in USA) web: http://www.a1.com/asm Fax: 317-254-8690 (If you experience difficulty in accessing our website, note that the web address uses numeral "1" in "a1")
We use a 1:2 mixture of OCT with our cryoprotectant buffer (PO4 + 20% sucrose) freezing in isopentane cooled in liquid nitrogen. This gives us a block consistency that allows for 3 um cryosections at -20 C, see Barthel & Raymond, J. Histochem Cytochem, 1990, 38:1383-1388 or our web site for a detailed protocol, http://www.personal.umich.edu/~praymond/ Others have used this combination with good success for both thin and thick (+10 um) sections.
To avoid sample cracking we try to be sure that ratio of block size to tissue size is large. It seems our tissue swells somewhat when it freezes. If the block is not large enough then it cracks. Sometimes we avoid the cracking by "quick" dipping the sample in the cooled isopentane until it is completely frozen.
If anyone has other suggestions to avoid cracking that would be great. It is a continuous battle with our samples. Seems some days are worse than others.
For using the samples for RNA (I presume you mean in situs?) we treat the samples no differently except that our reagents and labware (including poly-L-lysine slides) are RNase free.
When collecting your sample from the OR is it necessary to directly freeze there? Your best preservation would be if you placed your sample in a bottle of fix and returned to the lab to continue the processing. Linda Barthel Research Associate II Department of Anatomy and Cell Biology University of Michigan lab (313) 764-7476 fax (313) 763-1166 barthel-at-umich.edu
We use a 1:2 mixture of OCT with our cryoprotectant buffer (PO4 + 20% sucrose) freezing in isopentane cooled in liquid nitrogen. This gives us a block consistency that allows for 3 um cryosections at -20 C, see Barthel & Raymond, J. Histochem Cytochem, 1990, 38:1383-1388 or our web site for a detailed protocol, http://www.personal.umich.edu/~praymond/ Others have used this combination with good success for both thin and thick (+10 um) sections.
To avoid sample cracking we try to be sure that ratio of block size to tissue size is large. It seems our tissue swells somewhat when it freezes. If the block is not large enough then it cracks. Sometimes we avoid the cracking by "quick" dipping the sample in the cooled isopentane until it is completely frozen.
If anyone has other suggestions to avoid cracking that would be great. It is a continuous battle with our samples. Seems some days are worse than others.
For using the samples for RNA (I presume you mean in situs?) we treat the samples no differently except that our reagents and labware (including poly-L-lysine slides) are RNase free.
When collecting your sample from the OR is it necessary to directly freeze there? Your best preservation would be if you placed your sample in a bottle of fix and returned to the lab to continue the processing. Linda Barthel Research Associate II Department of Anatomy and Cell Biology University of Michigan lab (313) 764-7476 fax (313) 763-1166 barthel-at-umich.edu
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Not just an idea, but two GREAT ideas! I knew that the free spirits on the listserver would think of good stuff. Thanks. CS } } What about a piece of black and and white photographic negative film for a } cheap test specimen? On a good microscope, the silver grains appear with } sharp edges. I don't have a toy microscope around to compare. But film is } widely available and cheap, dry, and you don't even need a microscope slide } or coverslip. No regular pattern to detect barrel or pincushion distortion, } but these probably wouldn't matter to most users anyway. } } BTW, someone around here made up a bunch of cheap stage micrometers by } photographing a metric ruler from a distance with a 35 mm camera. With the } reduction, the one mm divisions on the scale are 100 micrometers apart on } the negative. These were then just cut up and glued to microscope slides. } They are a little fuzzy but if you measure center to center of each line it } is close enough. Perhaps one could photograph a grid to make a test } specimen for distortion. } } Gary Radice, Associate Professor gradice-at-richmond.edu } Department of Biology 804-289-8107 (voice) } University of Richmond VA 23173 804-289-8233 (FAX)
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
Yuhui wrote: ================================================== All of our scopes are JEOL made and we have been using filaments made by JEOL. The engineers from JEOL also recommend us to use their parts. As far as I know, however, there are companies which also sell filaments. Could anyone of you give me some information as to which source is better than the others? =================================================== While new filaments are always an option, have you considered retipping your left over filament bases? While not everyone would agree, there is a good number of persons who would claim that a good retipped filament is indistinguishable in its performance from a brand new one. Of course, not all retipped filaments from different sources are created equal. But if they work for you, a considerable amount of money can be saved.
Disclaimer: SPI Supplies offers a service to retip filaments as do others such as some of our competitors such as Pella, EBS, PLANO, etc. so we would all have a vested interest in seeing more people using retipped filaments instead of new ones. You can find out more information about the retipping of filaments from our website given below.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ =================================================
I have a sample that I have labeled with gold tagged antibody and I want to sputter coat it with aluminum for viewing in the SEM. We have a sputter coater with a turbomolecular pump and an aluminum target. We tired to sputter coat the sample using essentially the same "settings" used for chromium, but could not see that any coating had occurred. Does anyone have any ideas? Thanks in advance for any help any of you can give this humble student.
Dan - Never trust the low energy peaks from a specimen that is charging. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 400+ Links, MSDS ************************ http://www.proscitech.com.au
} Recently we did an SEM/EDX analysis that gave rather baffling results. } After some thought we came up with what seemed to be a plausible } explanation, however, the people for whom the analysis was done remained } skeptical. Therefore I am writing the Listserver for a second opinion. } The sample consisted of irregular shaped alumino-silicate particles roughly } 2-5 microns is size. The specimen was uncoated resulting in some charging. } Analysis (windowless) at 1000x mag. resulted in a carbon peak much larger } than the Al or Si peaks. At 5000x the carbon peak was smaller; i.e., } comparable to the Al and Si. Using a spot mode on individual particles, no } carbon was detected. However, spot mode analysis after lowering the } accelerating voltage from 20 to 10 kv, showed a small carbon peak. } The only explanation we could envision to fit these results was a thin } carbonaceous coating on the particle surface. Is this a plausible } explanation or we missing something? Does anyone have an alternative } suggestion? Any help would be appreciated. } } Dan Schwab } Center for Microscopy and Microanalysis } Bowling Green State University } Bowling Green, OH }
} How about an Environmental SEM? You could look at the sample in a fully } hydrated state, then slowly dehydrate, to reveal sub-surface structure. } } Regards, } Larry Stoter
We have successfully images hydrogel microspheres in the wet state using ESEM. See Applications of the environmental scanning electron microscope to the analysis of pharmaceutical formulations. D'Emanuele A, Gilpin C Scanning 1996 Oct 18:7 522-7
Chris
Chris Gilpin Biological Sciences Electron Microscope Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 161 275 5170 fax +44 161 275 5171
Is there a method of restricting a protozoa such as paramecium from moving about? The protozoa solution I have is great for studying them, but not if I want to look at the internal structure. Also what stain is a good stain to bring out the details in paramecium such as the nucleus?
Finally how do you pronounce 'paramecium'?!!!
Conrad :)
------------------------------------------------------------------------ ----------- "Any sufficiently advanced technology is indistinguishable from magic" ----------------------------------------------------------- Arthur C Clarke ----
I am working with the human cornea and wish to find the optimal buffers for glutaraldehyde and osmium tetroxide dilutions in the preparation of specimens for TEM. Any suggestions or references would be greatly appreciated.
} Cleaning grids? Go your hardest if you have nothing better to do: } Collect grids in a 20 or 30ml glass vial. Soak them in chloroform, give one } change in chloroform, pour off solvent, replace with ethanol or acetone,
Ethanol is better, if you store acetone and chloroform together in a waste bottle there is a risk, as those two will react explosively under some cirecumstances.
Also, after a final wash in aqueous media, if they are such as to remove the natural oxide coating, the copper underneath can oxidize and go all 'orrible.
} In a labour-costly country, with the odd exception, cleaning grids } is a waste of time.
I don't know if this applies to the USA, but in the UK we find that non-standard washing up is too difficult to be left to untrained staff!
+------------------------------------------------------------------------+ | 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 | +------------------------------------------------------------------------+
Yuhui Xu asked: } All of our scopes are JEOL made and we have been using filaments made by } JEOL. The engineers from JEOL also recommend us to use their parts. As far as } I know, however, there are companies which also sell filaments. Could anyone } of you give me some information as to which source is better than the others?
We (Energy Beam Sciences) have been manufacturing filaments for electron microscopes for more than 25 years. I know that some third party filaments are equal in quality to those purchased from the original equipment manufacturer, and some are not. In shopping for filaments, it is important to ask whether the filaments are vacuum-annealed (to minimize drift when heated) and precentered *after* annealing.
It is also important to know that, although the original equipment manufacturers all offer only one style of filament loop on their filament bases, usually a "V" hairpin filament (eg Philips) or a "curved" loop (eg JEOL), other filament loops can be mounted on the same bases to maximize performance in one way or another. For example, a pointed filament can dramatically increase brightness, while another loop configuration, such as our trademark "AR" loop, will provide longer life.
Anyone interested in the details of these special loop configurations can contact me directly back-channel.
Disclaimer: Energy Beam Sciences manufactures a wide range of tungsten filaments for electron microscopes.
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
I have had some similar samples generate noise in the spectrum which formed a peak in the area of carbon's. Usually the low energy side did not return toward baseline which was caused me to investigate a bit more. I found that if I used my Ultra Thin (metal/polymer) window the unusual response disappeared. If carbon was really there, the peak would be only slightly attenuated, but would head back "down hill" on the low side. I did no more experiments to determine the cause, but it was suggested to me by several that the sample was likely generating photons (~visible spectrum light) which were hitting the crystal - causing detector noise...
Regards, Woody White ______________________________ Reply Separator _________________________________
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello everybody,
Recently we did an SEM/EDX analysis that gave rather baffling results. After some thought we came up with what seemed to be a plausible explanation, however, the people for whom the analysis was done remained skeptical. Therefore I am writing the Listserver for a second opinion. The sample consisted of irregular shaped alumino-silicate particles roughly 2-5 microns is size. The specimen was uncoated resulting in some charging. Analysis (windowless) at 1000x mag. resulted in a carbon peak much larger than the Al or Si peaks. At 5000x the carbon peak was smaller; i.e., comparable to the Al and Si. Using a spot mode on individual particles, no carbon was detected. However, spot mode analysis after lowering the accelerating voltage from 20 to 10 kv, showed a small carbon peak. The only explanation we could envision to fit these results was a thin carbonaceous coating on the particle surface. Is this a plausible explanation or we missing something? Does anyone have an alternative suggestion? Any help would be appreciated.
Dan Schwab Center for Microscopy and Microanalysis Bowling Green State University Bowling Green, OH
Thats ok then, as that was how I did tend to pronounce it, I just thought it may have been pronounced:
PARA-MEK-EE-UM!!
I shall make a note of the useful info on the dyes! I currently have the standard methyl blue.
Conrad
} -----Original Message----- } From: Lou Ann Miller [SMTP:lamiller-at-uiuc.edu] } Sent: Wednesday, July 02, 1997 1:57 PM } To: Conrad Perfett } Subject: Re: keeping a protozoa still } } Pronouncing paramecium.... the US way: } } } pair a me see um } } } } I'm not an expert in the field, but I can tell you that ~ 0.5%( grams per } 100ml) Tolluidine Blue is often used for fungi. } } Crystal Violet along with iodine is often used for bacteria that's been } dried on a slide. } } Might try iodine, that would be inexpensive to try, maybe even a generic } Betadine at the drugstore / pharmacist's. } Would need a more dilute solution than what form it comes in, so as to not } overpower your object. } } } Also, speaking as one that has done about 10,000 hospital urine analysis, } try lowering the condensor by quite a bit, and it makes some things more } clear at about 40KX objective. ie floating blood cells, mucus strands, } casts , and a little protozoan called trichamonous. ( forgive my spelling } , it's been a while, and we just called them trich.) Which you hope you } never ever have! } } Also, 2 pieces of Xray film crossing at right angles ( polorized material) } will make the background darker, and make things like uric acid crystals } stand out. } } } Keep having fun! } } } } Lou Ann } } } } Finally how do you pronounce 'paramecium'?!!! } } } } Conrad :) } } Lou Ann Miller } Center for Microscopy & Imaging } College of Veterinary Medicine } Dept. of Veterinary Biosciences } University of Illinois } Rm 1108 Basic Sciences Bld } 2001 S Lincoln Ave. } Urbana, Illinois 61802 } } Phone: 217-244-1566 } email: lamiller-at-uiuc.edu } } Center for Microscopy & Imaging Home page: } http://www.cvm.uiuc.edu/MicImagLab/MicImagLab.html } } Central States Microscopy Society: } http://www.cvm.uiuc.edu/Homepages/LouAnnMiller/CSMS/csms.html } } Personal Home Pages: } http://www.cvm.uiuc.edu/Homepages/LouAnnMiller/LAM.html } }
How were the samples mounted during analysis? When we do EDX at our lab, we generally use clean, pure carbon pin-type stubs and carbon adhesive tape or discs. At a mag of 1000x while viewing particles 2-5 microns in size, it seems that your probe would be scanning a large area of whatever they were mounted on. If this is the case, going to higher mag and/or doing spot analysis, thereby eliminating more of the background, would be likely to reduce or eliminate the x-rays from the background, and this is what you observed.
Also, unless I'm mistaken, lowering the accelerating voltage would tend to emphasize the peaks of lower atomic weight elements, relative to heavier ones, so if you were getting x-rays from your mounting materials, they might tend to show up more at lower kv's.
Finally, if you're using a variable pressure scope, lower vacuum in the column can scatter the electron beam to a large degree. Again, this would tend to cause x-ray emission from areas other than the particles you are interested in.
Hope I'm not belaboring the obvious, but it really sounds to me like the carbon is from your mount.
Randy Tindall Center for Electron Microscopy Southern Illinois University at Carbondale
Dear microscopists, We will soon be purchasing an inverted metallograph for specimen prep. It appears all the major brands are quite good, but perhaps there are subtle advantages / disadvantages that are only evident after much use and experience. If anyone has any strong recommendations, I would appreciate your input. Please feel free to contact me off-line if you prefer. Thanks in advance for your time and help.
Sincerely, Mick Thomas
Materials Science Center Cornell University Ithaca, NY 14853 mgt3-at-msc.cornell.edu
At 6:59 PM 7/1/97, Linda Barthel wrote: see } Barthel & Raymond, J. Histochem Cytochem, 1990, 38:1383-1388 or our web } site for a detailed protocol, http://www.personal.umich.edu/~praymond/ } Others have used this combination with good success for both thin and } thick (+10 um) sections. } I tried to look up your web site and got an error message to the effect that the web site could not be found. Is this the right address?
thanks
Rob Homer
Robert Homer, MD, PhD Asst Prof, Pathology Yale University School of Medicine 310 Cedar St. PO Box 208023 New Haven, CT 06520-8023
Just a comment on a comment maninly on semantics...
} the cause, but it was suggested to me by several that the sample was likely } generating photons (~visible spectrum light) which were hitting the crystal } causing detector noise...
Visible light does not create noise, it is signal. Remember the SiLi detector is an ENERGY DISPERSIVE SPECTROMETER. The photons are energy packets just like X-rays both of which are creating electron/hole pairs. The energy is just such that is at the very low energy range of the detector and hence in correctly gets called noise. It may be signal you do not want to see, but it is signal. Preferential attenuation of the light by a very thin visible light absorbing "window" is a pretty standard trick to FILTER OUT the light (unwanted signal) from the low energy x-rays desired signal.
I guess that is one way! I did think of that, but I guess I wanted to keep it alive so I can see the processes going on! Although it may seem obvious, what is the best way to kill them? putting in something like a few drops of an alcohol solution?
Conrad
} -----Original Message----- } From: Robert Wieland [SMTP:wieland-at-me.udel.edu] } Sent: Wednesday, July 02, 1997 1:37 PM } To: Conrad Perfett } Subject: Re: keeping a protozoa still } } On Wed, 2 Jul 1997, Conrad Perfett wrote: } } [lots cut] } } Is there a method of restricting a protozoa such as paramecium from } } moving about? The protozoa solution I have is great for studying them, } } but not if I want to look at the internal structure. Also what stain is } } a good stain to bring out the details in paramecium such as the nucleus? } } There are any number of sources of methyl cellulose, which makes the } "water" they live in more viscous so they don't dart out of the field. To } make them absolutely still, I believe it's best to kill them. } } } } } Finally how do you pronounce 'paramecium'?!!! } } } par-ah-ME-see-um } } } } } Conrad :) } } } } ------------------------------------------------------------------------ } } ----------- } } "Any sufficiently advanced technology is indistinguishable from magic" } } ----------------------------------------------------------- Arthur C } } Clarke ---- } } } } } } Robert Wieland wieland-at-me.udel.edu } You can't go faster than light, you can't get colder than absolute } zero, and you can't help somebody by not telling them the truth. } }
Sorry about the web site address, there is one minor error, the correct address is: http://www-personal.umich.edu/~praymond/
On Wed, 2 Jul 1997, R Homer wrote:
} At 6:59 PM 7/1/97, Linda Barthel wrote: } see } } Barthel & Raymond, J. Histochem Cytochem, 1990, 38:1383-1388 or our web } } site for a detailed protocol, http://www.personal.umich.edu/~praymond/ } } Others have used this combination with good success for both thin and } } thick (+10 um) sections. } } } I tried to look up your web site and got an error message to the effect } that the web site could not be found. Is this the right address? } } thanks } } Rob Homer } } Robert Homer, MD, PhD } Asst Prof, Pathology } Yale University School of Medicine } 310 Cedar St. } PO Box 208023 } New Haven, CT 06520-8023 } } }
You don't mention what you mounted your sample on. If it is carbon based (carbon stub, conductive carbon tape, other adhesive using cargon), you will see the carbon peak. Going to smaller areas of excitation (i.e. increasing the relative proportions of sample to background scanned) will cause the carbon peak to be reduced. At spot mode on a particle, you are pretty much just exciting the particle and not the background so you do not see the carbon. At the lower accelerating voltage the emission volume may broden out closer to the surface of the background giving you the small carbon peak. It is important to remember that just because the initial point of excitation is small, it doesn't mean that the volume of exitation is small. With your sample, you are probably exciting several cubic microns of your sample. A good monte carlo program will demonstrate this to you.
Hope this helps
Michael D. Standing Electron Microscopist Brigham Young University e-mail: MDStandi-at-bioag.byu.edu
} I have a sample that I have labeled with gold tagged antibody and I want to } sputter coat it with aluminum for viewing in the SEM. We have a sputter } coater with a turbomolecular pump and an aluminum target. We tired to } sputter coat the sample using essentially the same "settings" used for } chromium, but could not see that any coating had occurred. Does anyone } have any ideas? Thanks in advance for any help any of you can give this } humble student. } } VTY, } Beth Bray } bbray-at-netside.com
Is it possible to successfull sputter Al? Al oxidises rapidly, so I would have thought you'd have a nice coating of Al oxide, an excellent insulator, on your specimen.
} I have a sample that I have labeled with gold tagged antibody and I want to } sputter coat it with aluminum for viewing in the SEM. We have a sputter } coater with a turbomolecular pump and an aluminum target. We tired to } sputter coat the sample using essentially the same "settings" used for } chromium, but could not see that any coating had occurred. Does anyone } have any ideas? Thanks in advance for any help any of you can give this } humble student.
I assume that you are coating with the lower atomic weight Al versus Cr in order to image the gold using backscattered imaging in the SEM. Lower atomic numbered metals are difficult to sputter coat properly - although it can be done using more energy. Also, Al is very reactive and will oxidize almost immediately after coating. Carbon would provide a better sputter coat if you have the capability. Actually, thermal evaporation is the preferred technique with both Al and C.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
In 1992, I did some SEM studies on corneal endothelial cells from rabbits. I found that a 0.1M phosphate buffered saline worked fine for that purpose. Osmolarity was maintained between 300 - 310. This work was published in the EMSA proceedings for that year; "Evaluation of the Biocompatibility of Polymer Surface Modifications with the Corneal Endothelium", p. 1106.
Regards,
Bob *************************** Bob Citron Chiron Vision Claremont, CA USA (909)399-1311 Bob_Citron-at-cc.chiron.com ***************************
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am working with the human cornea and wish to find the optimal buffers for glutaraldehyde and osmium tetroxide dilutions in the preparation of specimens for TEM. Any suggestions or references would be greatly appreciated.
Sorry for the incomplete info in my original posting. Several replies questioned the mounting method. The specimen was packed into small grooves in a silica-lead glass -- no carbon containing mountant of any kind was involved.
Thank you all for your expert advice and interest.
How about cubic zirconia for microscope objectives? RI = 2.15 - 2.45, dispersion = 0.060, birefringence = 0, hardness = 8.5 (quartz = 7), no cleavage with good toughness. Diamond is very hard, but has perfect cleavage in 4 directions (which means it chips relatively easily). So, CZ gets into diamond's range for RI, and should therefore make high NA lenses. CZ is a synthetic, so it can be grown under known conditions, and is relatively cheap and easy to produce. The high dispersion could be a problem for correcting chromatic aberration, but how much of one?
Phil
} Sic Hoc Legere Scis Nimium Eruditionis Habes { Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217) 355-1143 oshel-at-ux1.cso.uiuc.edu *** looking for a job again ******************
Beth, Al can be vey differcult to sputter off with DC process since it builds up a very thick, closely bonded native oxide layer on its surface. You might want to manually abrade the target to remove the oxide layer, pump down and sputter immeadiately. John Arnott
True... I was thinking in terms of undesired signal as noise. "Undesirable signal" is certainly more accurate!
Speaking of undesirable signal:
Some of my specimens have generated dead times as high as 90+ percent - with the beam OFF! Spectra (w/ beam on) have very wide peaks and a relatively huge background. I know the reason... Highly radioactive specimens wreak havoc with my EDS. Noise - or - signal???? {g}
BTW, I am using an (old) Kevex "Extra" detector. The massive turret snout does provide a bit of shielding. Would like to add more, but have no chamber room left (Etec).
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Just a comment on a comment maninly on semantics...
} the cause, but it was suggested to me by several that the sample was likely
} generating photons (~visible spectrum light) which were hitting the crystal
} causing detector noise...
Visible light does not create noise, it is signal. Remember the SiLi detector is an ENERGY DISPERSIVE SPECTROMETER. The photons are energy packets just like X-rays both of which are creating electron/hole pairs. The energy is just such that is at the very low energy range of the detector and hence in correctly gets called noise. It may be signal you do not want to see, but it is signal. Preferential attenuation of the light by a very thin visible light absorbing "window" is a pretty standard trick to FILTER OUT the light (unwanted signal) from the low energy x-rays desired signal.
HEEEEELP! I am a graduate student trying to get through my LAST expt and it is not working. I am coating copper grids with collodion (2% in amyl acetate) and trying to analyze mitochondrial dna on them. Initially this worked beautifully. The last 200 grids I've done have been a bust. Problems 1)the collodion keeps frying under the beam, it often seems to "wobble" under the beam. I have tried 3 different bottles of new collodion, cleaned the grids well with detergent-water-then acetone rinses all to no avail.
2)the dna seems to have stopped adhering to the collodion. I first heat nick the dna so it will be open circular (80 degrees, 30 min, in 0.5M NHAc) then I add cytochrome c to bind to the dna (I've tried concentrations from 2-100ug/ml) I then rotary shadow with platinum.
I do polarized light chemical microscopy in the style (or school) of Walter McCrone as part of doing general analytical chemistry for a pharmaceutical and chemical manufacturer. Examples of the types of problems I deal with are relating particle morphology of pure substances to properties and some rudimentary optical crystallography to distinguish crystal types. Can anyone provide experience or references on the possible usefulness of confocal or other "modern" light microscopic techniques in this area? Are there good journals covering such topics as well as classical PLM?
Dr. Leonard R. Corwin Principal Research Chemist Fort Dodge Animal Health Cyanamid Agricultural Research Center Quaker Bridge & Clarksville Roads PO Box 400 Princeton, NJ 08543-0400 609-716-2278 609-275-5239 fax corwinl-at-pt.cyanamid.com
Does anyone know where we can get our LKB 7800B knife breaker serviced? If possible, by someone in the Chicago area? It is making sporadically bad knives and we have adjusted all the knobs by the instruction booklet. If it needs to be replaced, can anyone recommend a good one? This one has been with us for over 20 years.
Thanks, Linda Fox, Loyola Univ. Medical Center, Chicago lfox1-at-wpo.it.luc.edu
To: Didier Le Thiec I.N.R.A. Centre de Recherches Forestieres Unite d'Ecophysiologie Forestiere Laboratoire de Pollution Atmospherique 54280 Champenoux - France
In response to request for information about WDX system with SEM or LVSEM.
Does anyone have a good SEM fixation method for planaria?
Nancy R. Smith Director of Operations Microscope And Graphic Imaging Center California State University, Hayward nsmith-at-csuhayward.edu http://www.csuhayward.edu/SCI/sem
} ... } } } I have a sample that I have labeled with gold tagged antibody and I } want to } } sputter coat it with aluminum ... } } ... } can be done using more energy. Also, Al is very reactive and will } oxidize } almost immediately after coating. ...
An Al oxidation layer tends to be very thin and impervious to further oxygen ... I might believe that under a vacuum the oxidization wouldn't occur until exposed to air, and would still be conductive (??)
cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
No, it's MEESE. And for a stain, go to a tropical fish store and get "ich" medecine. It's an aqueous copper salt & it works fine (see "Fun at 40 Power" in the MMMMMMProject MICRO bibliography).. Caroline
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
Try "Protoslo" quieting solution from Carolina Science and Math, Phone number: 1-800-334-5551. You might consider requesting a catalog. I believe that you might find they sell a number of interesting living and dead organisms for observation by light microscopy.
Regards,
Kirk J. Czymmek, Ph.D. Biological Electron Microscopy Facility 111 Wolf Hall University of Delaware Newark, DE 19716
Dear microscopists, We will soon be purchasing an inverted metallograph for specimen prep. It appears all the major brands are quite good, but perhaps there are subtle advantages / disadvantages that are only evident after much use and experience. If anyone has any strong recommendations, I would appreciate your input. Please feel free to contact me off-line if you prefer. Thanks in advance for your time and help.
Sincerely, Mick Thomas
Materials Science Center Cornell University Ithaca, NY 14853 mgt3-at-msc.cornell.edu
Dear Mick:
What would you be using this inverted metallographic microscope for? If you would use it for mostly tripod-type TEM sample preparation, I would highly recommend Lieca's model. It has a nice design and has higher resolution than other models that we've tested. If it is for general all purpose sample preparation and inspection, others would be more qualified to answer your question.
Regards,
Michael Coviello EM Lab Manager The University of Texas -at- Arlington Arlington, TX 817-272-5496
A week before our service contract for our Hitachi-7000 was to be renewed by Thomas Technical we were informed that they did not have the staffing for our TEM. We currently are totally without a contract. We cannot afford Hitachi's offer for the limited service we need. Can anyone recommend an independent company which we might contact? Any ideas would be appreciated. Thanks, Hildy Crowley Dept of Biol Sciences University of Denver 2101 E Wesley Ave Denver, CO 80208 Tel. 303-871-3026 E-mail { hcrowley-at-DU.edu } FAX 303-871-3471
The classic way to quite protozoa was a dilute solution of cocaine HCl. (Try explaining that to your local DEA officer) A solution of methyl cellulose will increase the viscosity of the mount to slow activity down for a good look. Years ago I tried Novocaine, which I obtained from my dentist, with limited success.
Best wishes...Frank ---------------------------------------------------------------------------- -------
These opinions are mine alone and have no relationship to my employer. Thank you.
Frank Karl fskarl-at-goodyear.com Goodyear Tire & Rubber Co. Voice: 330-796-7818 Analytical Services Dept. 415B Fax: 330-796-3304 142 Goodyear Blvd. Akron, OH 44305 USA
They that give up essential liberty to obtain a little temporary safety deserve neither liberty or safety. Benjamin Franklin
Is there a method of restricting a protozoa such as paramecium from moving about? The protozoa solution I have is great for studying them, but not if I want to look at the internal structure. Also what stain is a good stain to bring out the details in paramecium such as the nucleus?
Finally how do you pronounce 'paramecium'?!!!
We are teaching a summer course to high school kids using protozoans as our subject. They will be doing a lot of staining to show various internal structures, and a list of these stains is available on the "under construction" website for the course (http://131.229.114.77/LAL) - look under the section on examining protozoa. Most of the solutions are about .01-.05% aqueous. Takes some fussing around to get it right, as different types of protozoans vary a lot. There is a great text called "Protocols in Protozoology" edited by Lee and Soldo and published by the Society of Protozoologists. It has a lot of info on slowing 'em down - ranging from compression under the coverslip to various exotic techniques. Methylcellulose (10% aqueous, heating (don't boil it) to dissolve) is good. I have used CO2 (club soda water) added to the drop, as well as .01% nickel chloride for ciliates. Also try MgCl2 and KCl. Good luck. Dick Briggs
To: Didier Le Thiec I.N.R.A. Centre de Recherches Forestieres Unite d'Ecophysiologie Forestiere Laboratoire de Pollution Atmospherique 54280 Champenoux - France
In response to request for information about WDX system with SEM or LVSEM.
WDX requires a large probe current which means a large spot size and inferior resolution. Depending upon the specimen, the large beam current can produce specimen damage - not a problem for most bulky physical specimens but probably a problem for organic specimens. Cold stages can reduce damage for some specimens, but because damage is radiation damage breaking down molecules anot heat damage, the reduction in damage is not universal.
Operating in the Low Vacuum of Variable Pressure mode does not do anything to prevent or enhance the beam damage. the residual gas molecules will scatter the beam, resultiong in x-rays being generated at positions away from the major beam incidence region. For this reason, care should be exercised in interpreting the results obtained from x-ray studies in the variable Pressure Sem mode. Shorter working distances minimize this effect. I hope shortly to report on another technique for further minimizing this effect.
I can't give you references on WDX in variable pressure SEM, but following are some references for the early work on the development of the variable pressure SEM capability.
References on Variable Pressure SEM
V.N.E. Robinson, A Wet Stage Modification to a Scanning Electron Microscope - Proceedings of the Eighth International Conference on Electron Microscopy, Ed. J.V. Sanders and D.J. Goodchild, Australian Academy of Science, Canberra, Vol. II, pp 50-51.
V.N.E. Robinson, A Wet Stage Modification to a Scanning Electron Microscope, J. Microscopy, 103, pp 71-77, 1975.
V.N.E. Robinson, The Elimination of charging artifacts in the Scanning Electron Microscope, J. Phys. E: Sci. Instrum. 8, pp 638-640, 1975.
V.N.E. Robinson, Scanning Electron Microscope Environmental Cells, Scanning Electron Microscopy 1976/I, Ed. O Johari and I Corvin, IITRI, Chicago pp 91-100.
V.N.E. Robinson, The Examination of Hydrated Biological Specimens in a Scanning Electron Microscope Environmental Cell, Electron Microscopy, 1976, Proc. 6th European Congress, Ed. Y. Ben-Shaul, Tal International, Jerusalem. Vol. II, pp 85-90.
D.A. Moncrieff, V.N.E. Robinson and L.B. Harris, Charge Neutralisation of Insulating Surfaces in the SEM by Gas Ionisation, J. Phys. D: Appl. Phys. Vol 11, pp 2315-2325, 1978.
D.A. Moncrieff, P.R. Barker and V.N.E. Robinson, Electron Scattering by Gas in the Scanning Electron Microscope, J. Phys. D: Appl. Phys. Vol. 12, pp 481-487, 1979.
G.D. Danilatos and V.N.E. Robinson, Principles of Scanning Electron Microscopy at High Specimen Chamber Pressures, Scanning, Vol 2, pp 72-82, 1979.
V.N.E. Robinson, the Examination of Hydrated Specimens in electron Microscopes, in Analysis of Organic Biological Surfaces, John Wiley and Sons, New York, 1984, Ed. P. Echlin, Ch. 8, pp 191-207.
Tom Ruscica ETP USA, Electron Detectors Inc. For V.N.E. Robinson
Thanks to all of you who sent replies in answer to my question about sputter coating with aluminum. The consensus appears to be that, in general, "you can't get there from here." Aluminum was chosen because, as John Bozzola correctly assumed, I want to image the gold using backscattered imaging in the SEM. I should have included that little fact in my initial query. My next move will be to check into using Zn, Cr, Cu, Ag, or Pd--which was another suggestion, and to research some of the suggested articles, as well as looking at using thermal evaporation. There are so many things to learn--whew!--but I am having more fun than I would ever have thought possible. I am a "mature student" who is fortunate enough to work for a company that is paying my way back to college to finish a degree that I started many years ago. I will graduate in December of this year (God willing and the creek don't rise) and I am having a blast in school! Any further ideas will very much welcomed. Again, many thanks!
Dear microscopists, We will soon be purchasing an inverted metallograph for specimen prep. It appears all the major brands are quite good, but perhaps there are subtle advantages / disadvantages that are only evident after much use and experience. If anyone has any strong recommendations, I would appreciate your input. Please feel free to contact me off-line if you prefer. Thanks in advance for your time and help.
Sincerely, Mick Thomas
Materials Science Center Cornell University Ithaca, NY 14853 mgt3-at-msc.cornell.edu
REPLY:
Dear Mick:
What would you be using this inverted metallographic microscope for? If you would use it for mostly tripod-type TEM sample preparation, I would highly recommend Lieca's model. It has a nice design and has higher resolution than other models that we've tested. If it is for general all purpose sample preparation and inspection, others would be more qualified to answer your question.
Regards,
Michael Coviello EM Lab Manager The University of Texas -at- Arlington Arlington, TX 817-272-5496
} The classic way to quite protozoa was a dilute solution of cocaine HCl.
As was mentioned use methyl cellulose. It is sold under the name Protoslo. Another trick is to chill the Protozoa medium and to use neutral density filters to keep the heat of the lamp off of the little buggers.
Blystone in Texas
Robert V. Blystone, Ph.D. {RBLYSTON-at-Trinity.edu} Professor of Biology Trinity University San Antonio, Texas 78212 210.736-7243 210.736-7229 FAX
Sorry to be so long in geting to this. In the rat race of life the rats are winning. Now to split a few hairs with you Mark, and hopefully we will not bore the list to death or scare away those amateurs. On Tue, 1 Jul 1997, Dr. Mark W. Lund wrote:
} } Lenses have several types of aberrations which will cause the loss of } } detail unless corrected for. Toy microscopes are not. } } } } Spherical aberration - "is especially apparent in lenses having sperical } } surfaces. Light paths near the center of the lens focus at different } } points compared to light paths near the periphery." } } } The name "spherical aberration" has historical roots in astronomy where } a spherical mirror has this aberration but a parabolic mirror does not. } In some instances a spherical mirror will have no spherical aberration } wheras a parabolic mirror will have maximum spherical aberration. } The aberration really has nothing to do with the spherical surfaces } of lenses. The confusion comes because making a surface "aspheric" } can cure spherical aberration in many instances.
What confusion? Spherical aberation (also sometimes called aperture aberation) is a recognized term. While the refraction of light remains constant, the "distance traveled" in the lens and the angles of the lens to the object change as a function of the curvature of the lens surface. That curvature resembles the surface of a sphere.
} } } } Field curvature - "is a natural result of using lenses with curved } } surfaces. The image plane produced by such lenses will be curved. This } } kind of image occurs in microscopy unless plano (flat-field) objectives } } are used." } } Actually, field curvatureis a natural result of the geometry of the } real world. Since an object at the edge of the field of view is } farther from the lens "center" it will tend to be focussed closer to the lens } than an object on axis. This naturally leads to field curvature unless } the designer makes the lens weaker for off axis points. It has nothing } to do with the lenses being curved. The confusion comes from the formula } for Petzval curvature, which has lens powers in it.
And the geometry you refer to is due to what? I would suggest that it is due to the curved surface/s of the lens. The curvature of the image does mimic the curvature of the lens.
As to "off axis points" we are not getting into Coma aberration are we?
To those one the list, hopefully this friendly discussion/hair spliting session does not bore you all to tears.
as an alternative to having your knife-maker serviced have you considered replacing the scoring tool part. I am familiar with the LKB 7801A and it's a simple task to replace the cutting wheel (it should be in the instructions). If you haven't got any spares then you may have to trace a supplier but a pack of several replacements would cost you less than the call-out for a service. My experience with these knife makers is that they virtually go on for ever.
Sorry if you've already done this.
Malcolm Haswell e.m. unit University of Sunderland UK ----------
Beth:
An additional thought, why not coat with a thin layer of evaporated carbon. You would only need to put on a few nm.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Thanks to all of you who sent replies in answer to my question about } sputter coating with aluminum. The consensus appears to be that, in } general, "you can't get there from here." } Aluminum was chosen because, as John Bozzola correctly assumed, I want } to image the gold using backscattered imaging in the SEM. I should have } included that little fact in my initial query. My next move will be to } check into using Zn, Cr, Cu, Ag, or Pd--which was another suggestion, and } to research some of the suggested articles, as well as looking at using } thermal evaporation. } There are so many things to learn--whew!--but I am having more fun than } I would ever have thought possible. I am a "mature student" who is } fortunate enough to work for a company that is paying my way back to } college to finish a degree that I started many years ago. I will graduate } in December of this year (God willing and the creek don't rise) and I am } having a blast in school! } Any further ideas will very much welcomed. Again, many thanks! } } Beth Bray } bbray-at-netside.com } } }
As one fellow amateur to another, I'd be interested in the type of camera you are intending to use! I currently use a Praktica MTL50 but am thinking of getting an OM1 or 2. I have looked around and even secondhand they are still expensive! but it seems Olympus are the only ones that have the optionable clear screen (which seems to cost anywhere from 30 to 40 UK pounds for that alone!). I was thinking of following someones advice on here (from Australia if I remember correctly) and cementing a square coverslip with canada balsam to the focusing screen of another cheap camera, probably another Praktica as they are dirt cheap here secondhand. Anyone else tried this? Is it satisfactory? I realise it would make the camera only suitable for microphotography but that does not matter as it would be vastly cheaper.
Conrad
} -----Original Message----- } From: Frank J. Hogan [SMTP:jhogan-at-freenet.npiec.on.ca] } Sent: Sunday, June 29, 1997 4:16 AM } To: Conrad Perfett } Subject: RE: Amateur Microscopy } } Hi again Conrad, } Been away for a bit and just got caught up with the traffic on the } mic list....wow! you really started something good. It really is time } somebody looked into the toy microscope scene or should I say scam. I had } one when I was a little guy and it really put me off scoping until I got } my Bushnell lab model for med school. } I didn't graduate from med school but switched to journalism where I } earned my bread until recent early retirement. Fortunetely I kept my scope } after med school days. Now, I hope to make use of it as a hobby tool. } My scope has all the bells and whistles a lab scope should have I } guess. I am limited by a three objective turret, 4x,10x,40x and an 8x and } 10x eyepieces. I am equiped to do photomicroscopy and it is something I'm } looking forward too. } From the sound of things all you folks on the list are way ahead of me } equipment-wise, but I'm content to putt along with what i've got for now. } It also sounds like the more advanced people are providing all the info } you can handle. } } Very best } Frank }
I am a non-smoker! Also I *do* already have a protozoa solution to slow the critters down, but was looking more for a way to keep them still so I can look and photograph them
Thanks for the replies I now have a wealth of information!
Conrad :)
} -----Original Message----- } From: Keith Ryan [SMTP:KPR-at-wpo.nerc.ac.uk] } Sent: Thursday, July 03, 1997 9:07 AM } To: fskarl-at-goodyear.com; microscopy-at-Sparc5.Microscopy.Com } Subject: RE: keeping a protozoa still -Reply } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Conrad, You could try putting the little critters in the fridge for a while before you look at them. This works for fruit flies and slows them down, so it may be worth a try. Good luck Nikki
******************************** Nikki Bock EM Technician Dept.ME&MD University Of Nottingham Nottingham NG7 2RD Tel: (0115) 9513759 Email: emznjb-at-emn1.nott.ac.uk
I am re-posting my question, since I have not received any replies and our further trials are still unsuccessfull. Maybe this time, with a different Subject, somebody will notice and help us. My appologies to those not interested in immuno.
Dear immuno colleagues,
We would like to label F-actin at the EM level in growth plate chondrocytes and matrix vesicles. We have done some pilot experiments at the light microscope level using:
1) Rabbit anti-actin antibody (A 2066 from Sigma) on paraffin sections. The label was confined only to a subset of smooth muscle cells in control tissues.
2) Phalloidin-BODIPY FL worked very nicely on cryostat sections, therefore we tried anti-BODIPY antibody, followed by biotinylated goat-anti-rabbit, Streptavidin peroxidase and AEC substrate. However, we got no labelling.
We would appreciate any input on F-actin labelling at EM level in non-muscle cells or any experience dealing with rabbit anti-BODIPY FL antibody.
Thank you,
Sarka Lhotak EM Facility, McMaster University Hamilton, Ontario, Canada
I am re-posting my question, since I have not received any replies and our further trials are still unsuccessfull. Maybe this time, with a different Subject, somebody will notice and help us. My appologies to those not interested in immuno.
Dear immuno colleagues,
We would like to label F-actin at the EM level in growth plate chondrocytes and matrix vesicles. We have done some pilot experiments at the light microscope level using:
1) Rabbit anti-actin antibody (A 2066 from Sigma) on paraffin sections. The label was confined only to a subset of smooth muscle cells in control tissues.
2) Phalloidin-BODIPY FL worked very nicely on cryostat sections, therefore we tried anti-BODIPY antibody, followed by biotinylated goat-anti-rabbit, Streptavidin peroxidase and AEC substrate. However, we got no labelling.
We would appreciate any input on F-actin labelling at EM level in non-muscle cells or any experience dealing with rabbit anti-BODIPY FL antibody.
Thank you,
Sarka Lhotak EM Facility, McMaster University Hamilton, Ontario, Canada
I believe that Leica is now handeling (or has acquired) the LKB knife breaker. You may try to contact your local Leica sales rep. for more information.
Disclaimer: I have no personal or financial interest in Leica
Michael Standing Electron Microscopist Brigham Young University e-mail: MDStandi-at-bioag.byu.edu
---------- } From: NANCY SMITH {nsmith-at-gauss.sci.csuhayward.edu} } To: microscopy-at-Sparc5.Microscopy.Com } Subject: Planaria } Date: Mon, 30 Jun 1997 15:03:41 PSD8PDT } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I think this one works ok, it has been some years since I used it, but it was fine then! 200ml dist. H2O, 2 ml conc. HNO3, 4.5 ml formalin and 2.5 g MgSO4. Fix by dropping solution onto the planaria, done at room temp, a higher temp causes improper relaxation, a lower one, mucus secretion. Fixation complete within 24hrs, the specimens may be stored in 70% ethanol, or 5% formalin. Osmicate specimens and dehydrate as per usual. Good luck!
Alex Black Department of Anatomy University College Galway IRELAND
Further to my earlier note about tobacco smoke, I now have the reference: CFA OPantin. Notes on Microscopical Technique for Zoologists. Cambridge University Press. 1969., page 7.
Funnily enough, guess what it is recommended for - Paramecium !! Also flagellates, the cilia of Mytilus and Hydra. Not very politically correct (but in certain matters, I may not be!). Expedient, if not finally terminal for both the Paramecium and the microscopist!!
Also in this small book (of 76 pages) are recommended: 1. 10% alcohol - if you are an amateur, maybe whisky etc could be added to your water. 2. magnesium chloride - 7.5% MgCl2.6H2O or 20% MgCl2.7H20 are both isotonic with seawater, use half seawawater and half MgCl2 solution. This works atraet within minutes with baby cuttlefish and squid (in fact, we got a paper about it!)
Use 2.5% Mgcl2.6H2O fo freshwater organisms, it is slow e.g. 2 hours for Planaria.
3. Menthol. - scatter a few crystals on the surface of the water and leave overnight. Our specimen dept. used to do this with marine invertebrates.
4. Ether vapour - good luck (explosively flammable!)
A more modern item is polyethylene oxide with a Molecular Weight of 4,000,000. From the Aldrich catalogue, cat. no. 18,964-4, 5 grams costs #12.20 in UK before 17.5% tax. Used at 1% in your medium, this works very well. This is a tip from the person for whom I recently posted a fluorescence microscopy question.
Best wishes - Keith Ryan Plymouth Marine Lab., UK (Bonjour, Daniele!)
Does anyone know where we can get hydrophilic glass coverslips, please? We have seen a reference to them in a German paper, as supplied by ILMGLAS, but we cannot find this company on the web. Any other suppliers?
Thanks in advance,
+------------------------------------------------------------------------+ | 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 | +------------------------------------------------------------------------+
POSITION AVAILABLE: Laboratory Manager, Interdepartmental Laboratories
The American Museum of Natural History is seeking a Laboratory Manager to manage the Museum's Interdepartmental Laboratories. Duties include maintenance and operation of analytical microscopy equipment, particularly a scanning electron microscope with energy dispersive spectrometer, confocal laser scanning microscope, and digitial imaging and printing facilities; day to day management of laboratory operations; training users on analytical instrumentation; and participation in biological, geological and anthropological research. Qualifications include an advanced degree in Museum science-related field, 5 years experience in analytical laboratory operation and with scanning electron microscopes and energy dispersive systems. Ability to work with research scientists and to manage day-to-day operation of a digital imaging facility also required Background in confocal laser scanning electron microscopy desirable. Full Benefits. Please send resume with salary history to:
Human Resources American Museum of Natural History Central Park West at 79th Street New York, NY 10024-5192.
You may also e-mail your application to: barnett-at-amnh.org
An Equal Opportunity Employer. No phone calls or faxes, please.
-------------------------------------------------------------------------- | William K. Barnett, Ph.D. | | Director, Interdepartmental Laboratories barnett-at-amnh.org | | American Museum of Natural History | | Central Park West at 79th Street | | New York, NY 10024-5192 | --------------------------------------------------------------------------
The recent discussion about inexpensive, high quality microscopes, reminded me that I have a couple of excess microscopes that I would be happy to sell. These were my primary instruments until I recently moved up to Zeiss. They are older, Bausch and Lomb/American Optical Spencer microscopes with illuminators, excellent objectives, lenses, and condensors. They are in excellent condition with available lenses ranging from 3.5x to 100x and include oil immersion. If I remember right, eyepieces are either 6x or 10x. These are very good for amateur, exploration, etc. at a low cost. No plastic lenses or cheap parts.
Price: 150 U.S. to $250 U.S. (depending on what you want in the say of objectives and lenses) plus shipping.
I won't be able to answer you for a few days, but let me know what you think. I'll get back to you shortly.
We're redefining the way the world looks at life sciences. We're the new life sciences company at Monsanto. We're even more focused on the future. That's why our entire organization, from agricultural biotechnology to pharmaceuticals to food ingredients, is dedicated to life sciences. These are areas that can directly impact our future. Now, we can continuously answer the questions, needs and demands of our ever-changing planet. After all, the world's resources are finite, but our vision for its future is not.
Electron Microscopy Research Associate -
Monsanto Corporate Research has an opportunity for a Research Associate in electron microscopy. The position is located in the Analytical Sciences Center (ASC) in St. Louis, MO. The ASC is an exciting, state-of-the-art research organization serving the analytical needs of Monsanto's new life sciences company.
The successful candidate will join a small team of experts involved in the structural examination of catalysts, polymers, biomaterials, and other nanophase materials. The position requires an M.S. or B.S. or equivalent in material science or a related field with 3+ years experience in transmission electron microscopy (TEM) associated with materials. A strong background in TEM and associated preparative techniques, including embedding, ultramicrotomy, and dark room procedures is essential.
The successful candidate will be highly motivated, self-directed, interested in learning new skills, and effective working independently or in a team-based research environment. Excellent interpersonal, verbal and written communication skills are necessary. Experience with SEM and other imaging techniques, as well as digital imaging and image analysis are desired but not required.
Monsanto offers a competitive salary and benefits package, as well as a challenging and outstanding career opportunity in a fast-paced scientific organization. Interested applicants should send their CV or resume (suitable for scanning into our database), with the names and addresses of three references to: Monsanto Company, Staffing Code 0276, 800 N. Lindbergh Blvd., Mail Zone E2NA, St. Louis, MO 63167. EEO/AA Employer M/F/D/V. Please visit our website at www.monsanto.com.
} Aluminum was chosen because, as John Bozzola correctly assumed, I want } to image the gold using backscattered imaging in the SEM. I should have } included that little fact in my initial query. My next move will be to } check into using Zn, Cr, Cu, Ag, or Pd--which was another suggestion, and } to research some of the suggested articles, as well as looking at using } thermal evaporation.
Be careful in chosing the proper metal to coat your specimen because if you use a metal with too high atomic number (Pd, Ag, Au, Cr, etc) the backscatter emissions may mask completely the gold emission from the specimen - especially since it will be weak to begin with. In this case, thermally evaporated carbon is the coating of choice.
Isn't microscopy fun! Good luck in your career |8 { ))
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
I have been asked to study copper cathode microstructure with LM. Can anyone direct me to any descriptions, micrographs or papers written about cathode copper microstructure. Addresses of Companies and/or organizations to contact with for help on this subject would also be very helpful to me.
} HEEEEELP! I am a graduate student trying to get through my LAST } expt and it is not working. I am coating copper grids with } collodion (2% in amyl acetate) and trying to analyze mitochondrial } dna on them. Initially this worked beautifully. The last 200 } grids I've done have been a bust. Problems } 1)the collodion keeps frying under the beam, it often seems to } "wobble" under the beam. I have tried 3 different bottles of new } collodion, cleaned the grids well with detergent-water-then acetone } rinses all to no avail.
Check that you are not over-irradiating the specimen in the TEM (emission too high, too large condenser spot size or aperture, too low kV). OR you may have overloaded the grid with specimen material.
} 2)the dna seems to have stopped adhering to the collodion. I } first heat nick the dna so it will be open circular (80 degrees, 30 } min, in 0.5M NHAc) then I add cytochrome c to bind to the dna (I've } tried concentrations from 2-100ug/ml) I then rotary shadow with } platinum.
The cytochrome c may have gone bad. This happened to me once and the dna was not coated properly. Same batch used as previously?
Is the rotary shadowing being done properly - i.e., adequate Pt being deposited? If you put down enough Pt, this should stabilize the collodion as well. Not enough Pt will lead to lack of contrast (no dna seen) as well as drifting/unstable films.
Did you try checking out plain collodion coated grids, minus specimen? If bad, try a different vendor.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 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 Microscopy ListServer -- Sponsor: The Microscopy Society of America
An aqueous extract of tobacco leaf usually has enough nicotine to make them comatose. And if you're a classroom volunteer, it's a dramatic & definitely PC demonstration for the young folk... Caroline
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
We have a similar setup though we use our TN 5500/5600 on a microprobe. There is a flex module that enable you to choose to send data out one of the four ports on the back of your 5500. It is the +} DV command. +} DV 0 is the default which sends data out the printer port. We send data to +} DV 4 and print out formated information to a PC via a 9-pin cable and into comm port 1 or 2. This is 1200 baud communications so images take a long time. On the PC side, we have basic programs that capture what is comming in. There are programs for both data string (analyses) and image transfer. All are very limited but being able to transfer data out of the PDP is an absolute nec.
E-mail if you want more info.
Ciao for now, Ken
Kenneth JT Livi Department of Earth and Planetary Sciences 34th and Charles Streets Johns Hopkins University Baltimore, Maryland 21218 USA Phone: (410) 516-8342 Fax: (410) 516-7933 e-mail: klivi-at-jhu.edu
Hi! Just a note on tobacco juice. I was told that stuffing tobacco leaves on your socks will prevent leaches from crawling up your legs as you cross rivers. Any scientific truth in it? Anything to do with the nicotine?
K.M. Khoo
On Thu, 3 Jul 1997, Caroline Schooley wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } -----------------------------------------------------------------------. } } } } Further to my earlier note about tobacco smoke, I now have the } } reference: CFA OPantin. Notes on Microscopical Technique for } } Zoologists. Cambridge University Press. 1969., page 7. } } } } Funnily enough, guess what it is recommended for - Paramecium !! Also } } flagellates, the cilia of Mytilus and Hydra. Not very politically correct (but } } in certain matters, I may not be!). Expedient, if not finally terminal for } } both } } the Paramecium and the microscopist!! } } An aqueous extract of tobacco leaf usually has enough nicotine to make them } comatose. And if you're a classroom volunteer, it's a dramatic & } definitely PC demonstration for the young folk... } Caroline } } } } Caroline Schooley } Educational Outreach Coordinator } Microscopy Society of America } Box 117, 45301 Caspar Point Road } Caspar, CA 95420 } Phone/FAX (707)964-9460 } Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html } Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/ } } }
I travel the world teaching practical electron microscopy and it worries = me the emphasis that people place on "Spot Mode". I do not teach the use of=
spot mode for the following reasons-
1. Just because the spot appears in a certain position on the screen=
is this its correct position. I have found machines many microns out of step in X and Y directions.
2. Specimens charge and switching out of spot mode does not give the=
operator an easy opportunity to see if the spot had moved during the analysis.
3. Spot mode gives a false sense of analytical volume, no matter how=
small that spot may be we are almost certainly evaluating microns of material.
Do others worry about spot mode accuracy, do others test the spot mode accuracy? Is it not better to simply increase the magnification watching=
the area of interest all the way up before the analysis and all the way down after the analysis?
I have made an experiment with mites and than an embedding with Spurr=B4s epoxy resin. After polymerisation the blocks are fragile and shatter. Is there a possiblity to resolve Spurr=B4s and than to repeat the embedding of the samples or other solutions?
Thanks in advance
Heike Buecking Dr. Heike Buecking University of Bremen UFT Physiological Plant Anatomy Leobener Str. D 28359 Bremen Germany TEL: +49-421-218-2954 or TEL: +49-421-218-7283 FAX: +49-421-218-3737 e-mail: heibueck-at-uft.uni-bremen.de
unsubscribe =================================================================Dr. Francois Goutenoire Laboratoire des Fluorures - UPRES A 6010 Faculte des Sciences-Universite du Maine Avenuue O. Messian, BP 535 Fax:(33).2.43.83.35.06 72000 Le Mans Cedex, France Tel:(33).2.43.83.33.53 =================================================================
} Date: Fri, 04 Jul 1997 09:41:38 +0200 } To: microscopy-at-msa.microscopy.com } From: Heike Buecking {heibueck-at-uft.uni-bremen.de} } } Dear all, } } I have made an experiment with mites and than an embedding with Spurr=B4s epoxy resin. After polymerisation the blocks are fragile and shatter. Is there a possiblity to resolve Spurr=B4s and than to repeat the embedding of the samples or other solutions? } } Thanks in advance } } Heike Buecking Dr. Heike Buecking University of Bremen UFT Physiological Plant Anatomy Leobener Str. D 28359 Bremen Germany TEL: +49-421-218-2954 or TEL: +49-421-218-7283 FAX: +49-421-218-3737 e-mail: heibueck-at-uft.uni-bremen.de
I would agree with Stephen that it is important to emphasise the possible inaccuracies with using spot mode. However I would not go as far as saying don't use spot mode.
Spot mode can be useful for both quick looks to check compositions of a variety of particles in a field of view as well as for accurate microanalysis. Obviously with the latter it is important to be aware of the possibility of drift and to be sure of the position of the spot. Certainly with thin sections it is often possible to see the burn marks where the beam hit the sample. I tend to emphasise to students that when using spot mode the analysis will be a volume of 1-2 microns diameter and depth depending on sample and conditions. Even when using general rastering the same problem will be there.
This is just my opinion.
} I travel the world teaching practical electron microscopy and it worries me } the emphasis that people place on "Spot Mode". I do not teach the use of } spot mode for the following reasons- } } 1. Just because the spot appears in a certain position on the screen } is this its correct position. I have found machines many microns out of } step in X and Y directions. } } 2. Specimens charge and switching out of spot mode does not give the } operator an easy opportunity to see if the spot had moved during the } analysis. } } 3. Spot mode gives a false sense of analytical volume, no matter how } small that spot may be we are almost certainly evaluating microns of } material. } } Do others worry about spot mode accuracy, do others test the spot mode } accuracy? Is it not better to simply increase the magnification watching } the area of interest all the way up before the analysis and all the way } down after the analysis? } } What do you think?
************************************** * Pete Ainsworth * * Dept. Geology & Applied Geology * * Lillybank Gardens * * University of Glasgow * * Glasgow G12 8QQ * * e-mail: ainswort-at-geology.gla.ac.uk * * Tel : 0141 330 5505 (direct) * **************************************
} ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } ----------------------------- } -----------------------------------------. } } I travel the world teaching practical electron microscopy and it } worries me } the emphasis that people place on "Spot Mode". I do not teach the use } of } spot mode for the following reasons- } } 1. Just because the spot appears in a certain position on the } screen } is this its correct position. I have found machines many microns out } of } step in X and Y directions. } } 2. Specimens charge and switching out of spot mode does not give } the } operator an easy opportunity to see if the spot had moved during the } analysis. } } 3. Spot mode gives a false sense of analytical volume, no matter } how } small that spot may be we are almost certainly evaluating microns of } material. } } Do others worry about spot mode accuracy, do others test the spot mode } } accuracy? Is it not better to simply increase the magnification } watching } the area of interest all the way up before the analysis and all the } way } down after the analysis? } } What do you think?
Stephen - you are absolutely right - infact the only way to be sure where the spot was is to look for the specimen damage after the fact! If you have to analyze a small area (first think about the excitation volume) you are better off doing a redcued area scan or go to high magnification so you can at least "see" where the beam is hitting the sample.
I am looking for an electron diffraction analysis software package. I have to identify some particles (wether they're cubic, tetragonal, etc) and there must be an easier way than to hand draw the expected diffraction patterns....Does anybody know of something useful on the Web?
Thanx Sara
========================================================== Sara Prins Surface and Structure Analytical Services Division for Material Science and Technology CSIR PO Box 395 Pretoria SOUTH AFRICA
Depending on the stability of your stage/specimen, you should verify the position of the spot at required intervals. My collegues and I and numerous grad. students and post-docs have used this approach for decades.
On Fri, 4 Jul 1997, Bill Miller wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Stephen Chapman wrote: } } } ------------------------------------------------------------------------ } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America } } To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } ----------------------------- } } -----------------------------------------. } } } } I travel the world teaching practical electron microscopy and it } } worries me } } the emphasis that people place on "Spot Mode". I do not teach the use } } of } } spot mode for the following reasons- } } } } 1. Just because the spot appears in a certain position on the } } screen } } is this its correct position. I have found machines many microns out } } of } } step in X and Y directions. } } } } 2. Specimens charge and switching out of spot mode does not give } } the } } operator an easy opportunity to see if the spot had moved during the } } analysis. } } } } 3. Spot mode gives a false sense of analytical volume, no matter } } how } } small that spot may be we are almost certainly evaluating microns of } } material. } } } } Do others worry about spot mode accuracy, do others test the spot mode } } } } accuracy? Is it not better to simply increase the magnification } } watching } } the area of interest all the way up before the analysis and all the } } way } } down after the analysis? } } } } What do you think? } } Stephen - you are absolutely right - infact the only way to be sure } where the spot was is to look for the specimen damage after the fact! If } you have to analyze a small area (first think about the excitation } volume) you are better off doing a redcued area scan or go to high } magnification so you can at least "see" where the beam is hitting the } sample. } } Bill Miller } }
} I have made an experiment with mites and than an embedding with Spurr=B4s } epoxy resin. After polymerisation the blocks are fragile and shatter. Is } there a possiblity to resolve Spurr=B4s and than to repeat the embedding of } the samples or other solutions?
Spurr's is notoriously susceptible to moisture which, when present, results in a brittle block that shatters during trimming. The moisture may be absorbed from the atmosphere if one allows the uncapped specimens to sit overnight (presumably to evaporate propylene oxide) or, of course, it may not have been removed properly from the specimen during the dehydration stage. I suspect that your resin has absorbed moisture from the atmoshere. Check this out by polymerizing a block of plain resin. If it shatters, toss out any components that may have absorbed moisture and start over. Spurr's is a good resin but it is toxic, potentially carcinogenic and sometimes a pain. Take precautions against contact (nitrile gloves), inhalation (fume or exhaust hood) and properly dispose of the components by polymerizing them.
########################### Dr. John Bozzola, Director Center for Electron Microscopy Southern Illinois University Carbondale, IL 62901 Phone: 618-453-3730 =46ax: 618-453-2665 ###########################
Dear friends, I have some problem in interpreting my transmittance electron microscopy. Please, let me know if you can help me. Sincerely Soroush Sardari PhD student, Faculty of Pharmacy, Univ. of Alberta, Edmonton, Canada, T6G 2N8 Fax: (403) 492-1217
I routinely demonstrate to users of our JEOL 840 that they need to be careful of hysteris during scanning. We have TV rate, a rapid scan, and two slow scan modes on our SEM. I switch on the cross hair display at TV rate and select a point of interest. I then switch to rapid scan and have to recenter the cross hairs. I then switch to slow scan mode and recenter again. Then I ask them which is correct? Of course the slower scan is most nearly correct. I can demonstrate by watching the brightness LEDs as I move the spot across a bright feature.
Fortunately, we are not using spot mode, per se, all that much. Both our scopes now have digital imaging with point-and-shoot x-ray analysis. It is much easier locating the point(s) of interest. But users still need to be aware that there may some lag in the scan while recording the digital image. Since we tend to use rather slow scans that is not much of a problem. But users still need to be aware.
Depending on the rush, I still often use your method of cranking up the manification so that the feature fills the field of view. I am fairly assured that what I see is what I get for x-rays.
At 03:23 AM 7/4/97 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
One way to record experimental data on objects, rooms, and even reciprocal-space, may be to record images along directions evenly-distributed over the surface of a sphere. As Bucky Fuller pointed out in our neck of the woods some years ago, the 20 face-normals of an icosahedron are especially convenient in this regard, in part because the pictures required can be shot with a single 24-exposure roll of film.
To illustrate, HTML templates for exploring and animating data-sets on a Mathematica-drawn Klein bottle, and our Philips EM430ST in its new low-vibration site, are now on the web*. A technical note on the angles for taking such pictures is linked to each template set. Let me know if you decide to make available similar views from your own labs, or have other application-related questions we might help with.
} A major problems with scanning at high mag are either the beam is } blanked while the beam waits for "start frame" or "start line" sync } signals, thus count times are inaccurate ... or, if the beam isn't } blanked then the volume analyzed is weighted to the upper left corner } and left edge ...
These aren't the only alternatives. I can't speak for other EDS manufacturers, but our system links the scan generator and the X-ray pulse processor so that raster retrace and settling times are treated as "dead time". So the counting (live) time is correct and the pixel weightings are equal.
} 1. Just because the spot appears in a certain position on the screen } is this its correct position. I have found machines many microns out of } step in X and Y directions.
In my case, I'm always worried about this. I have, however, found that for every electron-opaque object I have measured I have found the ap- propriate element(s) when the beam is on the object and not when it is ~1 micrometer away. In many cases, there is a dark spot at the measured site after the measurement, but not before. Since these spots are at the position where the beam was set, and since occasional specimen drifts are accompanied be shifts of the spot and lowered or absent peaks (compared to subsequent measurements), I think I can be confident about the positions of my analyses. } } 2. Specimens charge and switching out of spot mode does not give the } operator an easy opportunity to see if the spot had moved during the } analysis. } A distinct advantage of the use of high voltage (~1MV) and very low beam current is that charging is very seldom a problem. Since I don't have scanning capability, it is of little concern if I have to take a long time to accumulate counts. If I were to do mapping, however, low beam current would be very detrimental.
} 3. Spot mode gives a false sense of analytical volume, no matter how } small that spot may be we are almost certainly evaluating microns of } material. } Especially in my case where the typical specimen is a biological section ~1 micrometer thick. David Joy's Monte Carlo program shows pretty well what the measurement volumes are.
} Do others worry about spot mode accuracy, do others test the spot mode } accuracy? Is it not better to simply increase the magnification watching } the area of interest all the way up before the analysis and all the way } down after the analysis?
Having complete control of the spot size by being able to set the condenser aperture and two condenser lenses independently of the magnifica- tion, I just use standard settings for each analysis--10K mag, 30 micrometer C2 aperture, maximally excited C1 lens, C2 to crossover. Yours, Bill Tivol
Dear Colleagues: Recently we came across a sample for SEM which we have great difficulty to process. We would like to ask for your help: This is a kind of poly-electrolyte polymer, that has been treated to form microspheres ( 1 to 10 micron in diameter ) and cross-linked to maintein the folded shape. Since the beads are highly negatively charged they do not stick to poly-L-lysine coated cover slips. We tried to run the pellets in Eppendorf tubes through alcohol dehydration, critical point drying, and then sprinkled the dried ( as powder) onto double-sided scotch tape and sputter coated. The results are generally not satisfactory partly due to the fact that it is very difficult to dry these beads thoroughly in tubes. I wonder whether there have been better methods for this type of sample which we do not know of. Any suggestions will be very much appreciated. Best regards, Yuhui Xu
} I travel the world teaching practical electron microscopy and it worries me } the emphasis that people place on "Spot Mode". I do not teach the use of } spot mode for the following reasons- } } 1. Just because the spot appears in a certain position on the screen } is this its correct position. I have found machines many microns out of } step in X and Y directions. } } 2. Specimens charge and switching out of spot mode does not give the } operator an easy opportunity to see if the spot had moved during the } analysis. } } 3. Spot mode gives a false sense of analytical volume, no matter how } small that spot may be we are almost certainly evaluating microns of } material. } } Do others worry about spot mode accuracy, do others test the spot mode } accuracy? Is it not better to simply increase the magnification watching } the area of interest all the way up before the analysis and all the way } down after the analysis? } } What do you think?
While the spot mode leaves a lot to be desired, particularly if the user isn't aware of the problems, analysis in raster mode is not without its difficulties. Most SEMs use some sort of of sychronisation of the line and frame scan signals. If so, the analysis obtained in raster mode is biased towards one edge and one corner of an area which may not even be identical with the image area - the beam 'waits' at the beginning of each line and each frame to synch, and the display may actually be blanked for a small distance after the scan starts and before the scan ends (over scanning) to hide image distortions arising from hysteresis, which is usually most evident at the edges of the scans.
Get a nice dirty specimen and scan it with a coarse raster until the contamination builds up. Now image the area at a lower mag, and look at the contamination pattern - there will probably be a heavier contamination line down one edge (the line scan 'wait') and a spot at one corner (the frame 'wait'). In addition, you may also be unlucky enough to find that the lines making up the coarse raster are bent at each end - you don't see the distortion in your images however because this part of the frame is blanked from display, but it will contribute to an EDX analysis.
More modern SEMs have reduced these problems, but I believe that they are still present in many.
I think that this demonstrates that whatever 'employers' might want, and manufacturers try to provide, for anything beyond the most basic EM, you need a skilled and trained operator with a full understanding of the instrumentation, and the time to fully check and calibrate the machine (who needs to be properly paid for undertaking a highly sophisticated and skilled job). Otherwise you get results that are at best doubtful and at worst wrong.
Malcolm Thomas wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear microscopists, } We will soon be purchasing an inverted metallograph for specimen prep. It } appears all the major brands are quite good, but perhaps there are subtle } advantages / disadvantages that are only evident after much use and experience. } If anyone has any strong recommendations, I would appreciate your input. } Please feel free to contact me off-line if you prefer. Thanks in advance } for your time and help. } } Sincerely, } Mick Thomas } } Materials Science Center } Cornell University } Ithaca, NY 14853 } mgt3-at-msc.cornell.edu Hi Mick,
Several of my customers have the new Leica. It is a low profile scope, provides easy access, very little stage drift and great amount of working distance. It is a new design and seems to be very popular. Call the 800 number for information for their number (800) 555-1212.
With regards to my recommendation for the LocTite 460, how did it perform?
Good Luck,
Sincerely,
Gary Liechty Allied High Tech Products 2376 E. Pacifica Place Rancho Dominguez, Ca 90220
800-675-1118 310-762-6808 Fax
Products for Materiallographic, SEM and TEM Sample Preparation
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I will soon need to get one or more new diamond knives for use with biological specimens. I haven't purchased a new knife for several years, and am curious about the quality of knives currently available. I'd appreciate hearing from anyone who has purchased and used a new knife in the last year or so.
Does anyone know of a method/stain that would localize glycogen in cryo sections of tissue (specifically goldfish neural retina) fixed with 4% paraformaldehyde? We are interested in the light level, not EM. Linda Barthel Research Associate II Department of Anatomy and Cell Biology University of Michigan lab (313) 764-7476 fax (313) 763-1166 barthel-at-umich.edu
You may simply put one drop of your sample onto a 0.45 u milipore filter or a normal stub and air dry it for 2-3 hours.
Good luck,
On Mon, 7 Jul 1997, yuhui xu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear Colleagues: } Recently we came across a sample for SEM which we have great difficulty to } process. We would like to ask for your help: } This is a kind of poly-electrolyte polymer, that has been treated to form } microspheres ( 1 to 10 micron in diameter ) and cross-linked to maintein the } folded shape. Since the beads are highly negatively charged they do not stick } to poly-L-lysine coated cover slips. We tried to run the pellets in Eppendorf } tubes through alcohol dehydration, critical point drying, and then sprinkled } the dried ( as powder) onto double-sided scotch tape and sputter coated. The } results are generally not satisfactory partly due to the fact that it is very } difficult to dry these beads thoroughly in tubes. I wonder whether there have } been better methods for this type of sample which we do not know of. Any } suggestions will be very much appreciated. } Best regards, } Yuhui Xu }
*********************************************** * Ming H. Chen, PhD * * Medicine/Dentistry Electron Microscopy Unit * * University Of Alberta. * * Edmonton, Alberta, Canada * * * * Visit My Page At: * * http://www.ualberta.ca/~mingchen * ***********************************************
I have been trying to identify ESEM EDS resolution using a 50 micron diameter Nickel standard embedded in epoxy (C, O and Cl). My instrument parameters are 1: 15kv 2: 12.0 mm working distance 3: Chamber pressure=3.5T 4: Chamber Gas = N2 or H2O (did'nt make any difference which gas was used) 5: Condensor setting = 50% 6: Sample tilt = 30, 20, 10. Doing a spot analysis in the center of the Ni standard showed abundant C and O, and some Cl as well as the expected Ni counts. Is the skirting effect of the ESEM such that one can't perform EDS on smples of 50 microns or less? I would be interested in hearing from anyone who has any results for ESEM EDS resolution.
PAS will stain glycogen but also other things in frozen sections. Can the sections be treated to reomve lipids? Otherwise, how about Best's Carmine? Give a day or two and I'll look in Lillie and Fullmer.
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 e-mail: mcauliff-at-umdnj.edu ***************************************************************
Responding to the message of {199707072115.AA24968-at-relay.ppco.com} from gllovel-at-ppco.com (Gary Lovell): } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I have been trying to identify ESEM EDS resolution using a 50 micron } diameter Nickel standard embedded in epoxy (C, O and Cl). My instrument } parameters are 1: 15kv 2: 12.0 mm working distance 3: Chamber } pressure=3.5T 4: Chamber Gas = N2 or H2O (did'nt make any difference which } gas was used) 5: Condensor setting = 50% 6: Sample tilt = 30, 20, 10. } Doing a spot analysis in the center of the Ni standard showed abundant C and } O, and some Cl as well as the expected Ni counts. Is the skirting effect of } the ESEM such that one can't perform EDS on smples of 50 microns or less? I } would be interested in hearing from anyone who has any results for ESEM EDS } resolution. } } From the literature on this subject I think that the chamber pressure is too high; resulting in too much scattering of the incoming beam. There are several papers by Brendon Griffin in Australia, and by the Danish group in RISO - Horsewell, Appel and Bilde-Sorensen - where they have extensively documented the EDS resolution as a function of voltage, pressure and working distance. See MSA proceedings from 1995 and 1996 for papers from the ESEM symposia which cover this topic
Good luck,
__________________ Stuart McKernan stuartm-at-tc.umn.edu Microscopy Specialist CIE Characterization Facility, University of Minnesota Phone: (612) 626-7594 100 Union Street S. E., Minneapolis, MN 55455 Lab: (612) 624-6590
In my undergrad. zoology lab long ago we added 7-Up to slow down marine invertebrates. Essentially they are a bit oxygen starved from all the CO2. Nicotine would cause larger organisms, such as chitons, to curl and distort.
} } } } Further to my earlier note about tobacco smoke, I now have the } } reference: CFA OPantin. Notes on Microscopical Technique for } } Zoologists. Cambridge University Press. 1969., page 7. } } } } Funnily enough, guess what it is recommended for - Paramecium !! Also } } flagellates, the cilia of Mytilus and Hydra. Not very politically } correct (but } } in certain matters, I may not be!). Expedient, if not finally terminal } for } } both } } the Paramecium and the microscopist!! } } An aqueous extract of tobacco leaf usually has enough nicotine to make } them } comatose. And if you're a classroom volunteer, it's a dramatic & } definitely PC demonstration for the young folk... } Caroline } } Glen MacDonald Virginia Bloedel Hearing Research Center Box 35-7923 University of Washington Seattle, WA 98195-7923 (206) 616-4156 glenmac-at-u.washington.edu *---------------------------------------------------------------------* The box said "Requires Windows 95 or better.", so I bought a Macintosh. *---------------------------------------------------------------------*
Hi Guys, I'm currently acquiring mosaic images of muscle fibers with Mosaic Tiling under Oncor Image version 2.0.5d. However, the copy of Mosaic Tiling that I have doesn't allow me to get high resolution mosaic image. Does anyone know if there is a copy of Mosaic Tiling for Oncor or anyother alternative that will allow me to get high resolution mosaic images good for quantification. Thanks,
--Ciprian Have fun and keep the sun on your back and a smile on your face. __________________________________________________________ Ciprian A. Almonte University of Pittsburgh Center for Biologic Imaging Pittsburgh, PA 15261
Visit my web site at http://www.pitt.edu/~calmonte Laboratory's website: http://sbic6.sbic.pitt.edu __________________________________________________________
Dear Roadwalk, The main difference between a biological and metallurgical light microscope is that the metallurgical microscope views in the reflection mode, where the light comes through the objective lens and reflects off the opaque sample, since very few metal samples are transparent to light. Many metallurgical microscopes are inverted, that is, the sample is put upside down on a stage on the top of the microscope, with the objective nosepiece underneath, so there is very little restriction on sample size. Several of our microscopes are dual purpose and can work in either transmitted or reflected light mode. They look like an ordinary biological microscope.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Regards, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
We would like to examine a pore size of cellulose acetate with SEM. Please advise us, how to prepare the sample becuase it is very sensitive to vacuum. Can we dry by using a CPD?
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Linda, Periodic Acid Schiff is first favourite. Any histochemistry text will give you the necessary details, failing that, e-mail me and I can send you my protocol. Ian.
well I now know why I am having difficulty finding the Amoeba since according to the book I have just bought, at least the classic Amoeba Proteus is rare in the wild!!! I remember being told by a retired microscopist that it was rare also. So at least I can take comfort in the fact that its more than likely there are none in my samples rather than my mis-identification! But I will not be giving up the search as everyone needs a mission in life *laugh*
Conrad
------------------------------------------------------------------------ ----------- "Any sufficiently advanced technology is indistinguishable from magic" ----------------------------------------------------------- Arthur C Clarke ----
I am trying to attatch various small samples (Diatoms, Microspores and Cecaria) onto Poly-L-lysine coated coverslips, but am finding that the samples do not attach.
Does anyone have any ideas?
My Method
Clean coverslips in 100% ethanol Air dry 0.1% Poly-L-lysine (MW60000) in distilled water for 1minute Rinse in distilled water Drop of fixative containg sample (30 - 60 minutes) rinse, dehydrate and CPD
I also remember reading somewhere about using Poly-D-lysine instead, has anyone tried this?
Thanks
Kevin Mackenzie Tillydrone E.M. Unit University of Aberdeen Tillydrone Avenue Aberdeen AB9 2NT
Tel 01224-272847 Fax 01224-272396 Web site- http://www.abdn.ac.uk/~nhi691/
} Responding to the message of {199707072115.AA24968-at-relay.ppco.com} } from gllovel-at-ppco.com (Gary Lovell):
} } I have been trying to identify ESEM EDS resolution using a 50 micron } } diameter Nickel standard embedded in epoxy (C, O and Cl). My instrument } } parameters are 1: 15kv 2: 12.0 mm working distance 3: Chamber } } pressure=3.5T 4: Chamber Gas = N2 or H2O (did'nt make any difference which } } gas was used) 5: Condensor setting = 50% 6: Sample tilt = 30, 20, 10. } } Doing a spot analysis in the center of the Ni standard showed abundant C and } } O, and some Cl as well as the expected Ni counts. Is the skirting effect of } } the ESEM such that one can't perform EDS on smples of 50 microns or less? I } } would be interested in hearing from anyone who has any results for ESEM EDS } } resolution. } } } } } } From the literature on this subject I think that the chamber pressure is too } high; resulting in too much scattering of the incoming beam. There are several } papers by Brendon Griffin in Australia, and by the Danish group in RISO - } Horsewell, Appel and Bilde-Sorensen - where they have extensively documented the } EDS resolution as a function of voltage, pressure and working distance. See MSA } proceedings from 1995 and 1996 for papers from the ESEM symposia which cover } this topic } } Good luck, } } Our group in Cambridge have also been investigating this, and will be presenting a paper at this years MSA conference in Cleveland :-
'Variations in the probe beam broadening with operating conditions in ESEM'
Tuesday 11:45 - Rm. 206
Ian Bache Research Student Polymers & Colloids Group, Cavendish Laboratory Cambridge University, Madingley Road, Cambridge CB3 0HE UK
SIGEE, D.C. and GILPIN, C.J. (1994) X-ray microanalysis with the environmental scanning electron microscope: Interpretation of data obtained under different atmospheric conditions. Scanning Microscopy Supplement 8: 219-229
1 GILPIN, C.J. and SIGEE, D.C. (1995) X-ray microanalysis of wet biological specimens in the environmental scanning electron microscope 1. Reduction of specimen distance under different atmospheric conditions. Journal of Microscopy 179: 22-28
Chris
Chris Gilpin Biological Sciences Electron Microscope Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 161 275 5170 fax +44 161 275 5171 http://www.biomed.man.ac.uk/biology/emunit/emhome.html
We do both theoretical and experimental work on high pressure SEM microscopy . (This is a noncommercial name for ESEM.) We have been looking at the effect of water vapour and water liquid layer on the emission characteristic x-rays. We have recently deduce from our Monte Carlo calculations that the true contribution from the 'image' area can be estimated by applying a correction factor. 'Resolved image area' in turn depends upon the thickness of the water layer or the pressure of the water vapour above the specimen. The exact proportion of the characteristic x-rays coming from the actual region being analysed increases with the 'increase' in the resolavable radius. Thus the value of the correction factor reduces as the resolution worsens. These results and others will be presented at the Meeting MSA.
Jitu Shah H.H. Wills Physics Laboratory, University of Bristol Royal Fort. Bristol BS 8 1TL UK email: jss-at-siva.bristol.ac.uk
} Stuart McKernan wrote: } } } Responding to the message of {199707072115.AA24968-at-relay.ppco.com} } } from gllovel-at-ppco.com (Gary Lovell): } } } } I have been trying to identify ESEM EDS resolution using a 50 micron } } } diameter Nickel standard embedded in epoxy (C, O and Cl). My instrument } } } parameters are 1: 15kv 2: 12.0 mm working distance 3: Chamber } } } pressure=3.5T 4: Chamber Gas = N2 or H2O (did'nt make any difference which } } } gas was used) 5: Condensor setting = 50% 6: Sample tilt = 30, 20, 10. } } } Doing a spot analysis in the center of the Ni standard showed abundant C and } } } O, and some Cl as well as the expected Ni counts. Is the skirting effect of } } } the ESEM such that one can't perform EDS on smples of 50 microns or less? I } } } would be interested in hearing from anyone who has any results for ESEM EDS } } } resolution. } } } } } } } } } From the literature on this subject I think that the chamber pressure is too } } high; resulting in too much scattering of the incoming beam. There are several } } papers by Brendon Griffin in Australia, and by the Danish group in RISO - } } Horsewell, Appel and Bilde-Sorensen - where they have extensively documented the } } EDS resolution as a function of voltage, pressure and working distance. See MSA } } proceedings from 1995 and 1996 for papers from the ESEM symposia which cover } } this topic } } } } Good luck, } } } } Ian Bache wrote: } Our group in Cambridge have also been investigating this, and will be } presenting a paper at this years MSA conference in Cleveland :- } } 'Variations in the probe beam broadening with operating conditions in } ESEM'
We do both theoretical and experimental work on high pressure SEM microscopy . (This is a noncommercial name for ESEM.) We have been looking at the effect of water vapour and water liquid layer on the emission characteristic x-rays. We have recently deduce from our Monte Carlo calculations that the true contribution from the 'image' area can be estimated by applying a correction factor. 'Resolved image area' in turn depends upon the thickness of the water layer or the pressure of the water vapour above the specimen. The exact proportion of the characteristic x-rays coming from the actual region being analysed increases with the 'increase' in the resolavable radius. Thus the value of the correction factor reduces as the resolution worsens. These results and others will be presented at the Meeting MSA.
Jitu Shah H.H. Wills Physics Laboratory, University of Bristol Royal Fort. Bristol BS 8 1TL UK email: jss-at-siva.bristol.ac.uk
} Stuart McKernan wrote: } } } Responding to the message of {199707072115.AA24968-at-relay.ppco.com} } } from gllovel-at-ppco.com (Gary Lovell): } } } } I have been trying to identify ESEM EDS resolution using a 50 micron } } } diameter Nickel standard embedded in epoxy (C, O and Cl). My instrument } } } parameters are 1: 15kv 2: 12.0 mm working distance 3: Chamber } } } pressure=3.5T 4: Chamber Gas = N2 or H2O (did'nt make any difference which } } } gas was used) 5: Condensor setting = 50% 6: Sample tilt = 30, 20, 10. } } } Doing a spot analysis in the center of the Ni standard showed abundant C and } } } O, and some Cl as well as the expected Ni counts. Is the skirting effect of } } } the ESEM such that one can't perform EDS on smples of 50 microns or less? I } } } would be interested in hearing from anyone who has any results for ESEM EDS } } } resolution. } } } } } } } } } From the literature on this subject I think that the chamber pressure is too } } high; resulting in too much scattering of the incoming beam. There are several } } papers by Brendon Griffin in Australia, and by the Danish group in RISO - } } Horsewell, Appel and Bilde-Sorensen - where they have extensively documented the } } EDS resolution as a function of voltage, pressure and working distance. See MSA } } proceedings from 1995 and 1996 for papers from the ESEM symposia which cover } } this topic } } } } Good luck, } } } } Ian Bache wrote: } Our group in Cambridge have also been investigating this, and will be } presenting a paper at this years MSA conference in Cleveland :- } } 'Variations in the probe beam broadening with operating conditions in } ESEM'
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Why would a silica shell of a diatom stick to a positively charge surface---- You can collect diatoms on a 0.22 micron or 0.45 micron silver filter sold by the electron microscopy houses. These filters are conductive if you are going to look at the diatoms in SEM.
George C. Ruben Dept. Biological Sciences Dartmouth College Hanover, NH USA
Mr-Received: by mta FWVX01; Relayed; Tue, 08 Jul 1997 08:37:27 -0400 Alternate-Recipient: prohibited Disclose-Recipients: prohibited
Gary,
I noticed a couple of things about your conditions which are probably not optimum for the best resolution. I assume you are using one of the Electroscan instruments. If so, you can reduce the beam skirt by using the long assembly secondary electron detector (17 mm) and a working distance of 19 mm. That leaves a 2 mm gas path length. Also, the voltage to the secondary detector has a profound effect on many of the measurement parameters such as the beam current (see Wight in 1996 MSA proceedings p838-839). Reducing that voltage probably has a beneficial effect As was mentioned previously, the size of the skirt is directly related to the gas pressure and one wants to operate as low as possible for the samples being examined. I use 2.0 torr for my polymer samples and lower for others. Also, higher accelerating voltages reduce the skirt size but at an obvious cost if doing EDS of light elements.
Notice how cleverly I gave an answer without answering your question? I should have been a lawyer or better a politician! Frankly, I don't think the final word has been written on EDS-ESEM resolution. The original work seemed to indicate that we should be talking millimeters not micrometers. More recent work, however, suggests that maybe micrometers are appropriate with sufficient care. There is a session on Tuesday morning on the ESEM at the MSA/MAS meeting and there will surely be more discussion of this issue there and in the literature.
By the way, I have been trying to locate the articles by Horsewell, Appel and Bilde-Sorenson mentioned by Stuart McKernan with no success. Does anyone have any suggesstions on how to get ahold of them?
Dear Yuhui; For particulate samples that cannot be air dried, I sandwich the sample between two Millipore filters held in a Millipore Swinney stainless steel filter holder. It is designed to fit on a syringe, but I cut the ends off (which allows for good fluid exchange), and run the holder through the dehydration series and CPD. Alternatively, some samples can be dried in HMDS or TMS, and you can skip the critical point dryer. Also, I recomend using conductive carbon tabs for your adhesive. They give a nice smooth background, and are more conductive than double-stick tape (mine came from Pella, but other companies may also offer them). Please feel free to contact me off-line for more details if necessary.
Leslie Eibest Zoology Dept., Box 90325 Duke University Durham, NC 27708 USA (919) 684-2547 leibest-at-duke.edu
} Assuming there is a device built on a 1cm by 1cm Si wafer and the } device has several 1-2nm thick layers and about 0.2um wide circuit } "wiring", how can you find failure points, preferably without breaking } the device? I know Sandi is doing X-TEM of devices and am really curious } about the way to find, say, a short circuit on the device. } Any comments are greatly appreciated. } Dear Chao-Ying, What are the compositions of the parts of the device you want to examine? That is, are the "wires" also mainly Si, are they a different material with similar Z--i.e. aluminum--or do they have much different Z? What I'm really asking is if there is a contrast-producing effect in the device. If all the device is Si with small amounts of various dopants for the different parts, you will have a difficult time, but if not, either imaging or element mapping could give you the info you want. Other questions are what resolution is necessary and what is the total thickness of the device--is there a thick backing? Yours, Bill Tivol
I have worked in biological EM for 13 years now, but am curious about metallurgical EM in Eng. and am wondering how difficult it would be for me to make a switch, because I've noticed that there seems to be more jobs for EM technologists in metallurgy than biology.
Are there any people out there that have switched from one side to the other? I think that looking after the microscope would be essentially the same, but I have no experience in electro-polishing and sample preparation for metallurgy. How difficult is this?
I am interested in hearing from anyone doing diagnostic EM with a company called MDS. We are about to be privatized here, but I have no idea what this company plans to do with electron microscopy. All the information that I have regarding MDS basically only applies to their high volume "core-lab" but not to their small specialized labs.
Responding to the message of {01IKZKXJZCKW90OHU4-at-mr.fwvx03.com} from "Robert A. CARLTON 610-454-3949" {CARLTRA-at-rpr.rpna.com} : } } By the way, I have been trying to locate the articles by Horsewell, } Appel and Bilde-Sorenson mentioned by Stuart McKernan with no success. } Does anyone have any suggesstions on how to get ahold of them? } The references I have are all to conference proceedings: MSA 1996 p847, Scandem 1996 Aarhus Denmark, Scandem 1997 Goteborg Sweden (and EUREM 1996 Dublin Ireland - proceedings on CR-ROM; incomplete and virtually useless!)
__________________ Stuart McKernan stuartm-at-tc.umn.edu Microscopy Specialist CIE Characterization Facility, University of Minnesota Phone: (612) 626-7594 100 Union Street S. E., Minneapolis, MN 55455 Lab: (612) 624-6590
} Date: Tue, 8 Jul 1997 08:51:14 +0100 (BST) } From: Kevin Mackenzie {nhi691-at-abdn.ac.uk} } To: Microscopy-at-sparc5.microscopy.com } Subject: Poly-L-lysine problem } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hi } } I am trying to attatch various small samples (Diatoms, Microspores and } Cecaria) onto Poly-L-lysine coated coverslips, but am finding that the } samples do not attach. } } Does anyone have any ideas? } } My Method } } Clean coverslips in 100% ethanol } Air dry } 0.1% Poly-L-lysine (MW60000) in distilled water for 1minute } Rinse in distilled water } Drop of fixative containg sample (30 - 60 minutes) } rinse, dehydrate and CPD } } I also remember reading somewhere about using Poly-D-lysine instead, } has anyone tried this? } } Thanks } } Kevin Mackenzie } Tillydrone E.M. Unit } University of Aberdeen } Tillydrone Avenue } Aberdeen } AB9 2NT } } Tel 01224-272847 } Fax 01224-272396 } Web site- http://www.abdn.ac.uk/~nhi691/ } SAMPLES THAT ARE FIXED FREQUENTLY DON'T LIKE TO STICK TO ANYTHING ELSE. TRY COATING YOUR SURFACE (WE USED 1% PL LYS FOR 10 MIN, RINSE, AIR-DRY), ADDING YOUR SAMPLE (LET A ROUNDED DROP SIT AND DON'T LET IT OVERFLOW THE EDGE) ON THE COVERSLIP FOR 30 MIN COVERED IN A MOIST CHAMBER. THEN REMOVE THE EXCESS FLUID WITH A PASTEUR PIPET AND *!*GENTLY*!* ADD FIXATIVE TO ONE SIDE AND LET IT SIT IN A ROUNDED UP DROP FOR 10-20 MIN. WASH WITH BUFFER (GENTLY) AND PROCEED WITH WHATEVER ELSE YOU WANT TO DO.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
} I have been trying to identify ESEM EDS resolution using a 50 micron } diameter Nickel standard embedded in epoxy (C, O and Cl). My } instrument parameters are 1: 15kv 2: 12.0 mm working distance 3: } Chamber pressure=3.5T 4: Chamber Gas = N2 or H2O (did'nt make any } difference which gas was used) 5: Condensor setting = 50% 6: } Sample tilt = 30, 20, 10. Doing a spot analysis in the center of the } Ni standard showed abundant C and O, and some Cl as well as the } expected Ni counts. Is the skirting effect of the ESEM such that } one can't perform EDS on smples of 50 microns or less? I } would be interested in hearing from anyone who has any results for } ESEM EDS resolution.
Dear Gary,
Approximately 75 % of the primary electrons will be scattered when you use a working distance of 12 mm and a pressure of 3.5 torr and a significant fraction of these scattered electrons will hit the sample further away from the beam target than 50 micrometer. (The scattered intensity is approximately given by Is/Io = 1 - exp(-psL/kT) where p is the pressure, s the total scattering cross section for electron scattering on the gas used, L the distance between the last pressure limiting aperture and the sample, k the Boltzmann constant and T the absolute temperature). Examples of skirt shapes are e. g. given in:
D. A. Moncrieff et al., J. Phys. D: Appl. Phys. vol. 12 (1979) 481-88. D. C. Joy, Microscopy and Microanalysis ' 96, Proc. Annual Meeting MSA, Minneapolis, Minnesota, 11 - 15 August 1996
It is to a large degree possible to correct for the beam skirt effects:
1) You can extrapolate from spectral measurements made at several different pressures to the result that would have been found without scattering provided that the measurements are made in the single scattering regime (i.e. pL { approx. 1.6 Pa.m for measurements in water vapour). In order to obtain single scattering conditions you can use a so-called X-ray bullet to reduce the working distance. 2) If there is plural scattering, you can take two spectra, one with a fine needle (of the kind used for field ion microscopy or scanning tunneling microscopy) inserted over the point of interest, and the other with the needle slightly retracted. Subtraction of the first from the second spectrum will approximately give the spectrum from the point of interest.
Neither method will give you as exact an analysis as you will get under high vacuum, but you can get rid of most of the skirt effects. The pressure variation method in particular yields pretty good results if carefully performed. The methods are described in:
J. B. Bilde-Soerensen and C. C. Appel, Proc. 48th Annual Meeting of the Scandinavian Society for Electron Microscopy, Aarhus, 2 - 5 June 1996, pp. 4 - 5. J. B. Bilde-Soerensen and C. C. Appel, Proc. 11th European Congress on Microscopy EUREM' 96, Dublin, 26-30 August 1996. Session T6. J. B. Bilde-Soerensen and C. C. Appel, Proc. 49th Meeting of the Scandinavian Society for Electron Microscopy, Gothenburg, 10 - 13 June 1997, pp. 12-15.
Best wishes, Jorgen.
J. B. Bilde-Soerensen Materials Research Department Risoe National Laboratory DK-4000 Roskilde Denmark
Job Title: TEM Analyst Manager: Dr. Carolyn Gondran Department: Materials Analysis Division: Internal Technical Support - SEMATECH 2706 Montopolis Dr. Austin, TX 78741-6499
(512) 356-3149 phone (512) 356-7008 FAX
Job Summary: Provide analytical support to SEMATECH and I300I projects and to the ATDF through TEM analysis of semiconductor materials and devices (in plan view and cross section) and direct interactions with internal customers. Operate the TEM (EDAX, STEM, PEELS, Electron diffraction etc.) interpret electron micrographs, electron diffraction patterns and EDAX data and provide written reports of all analyses. Work with and oversee the activities of TEM technician(s) including selection of sample preparation techniques, preparation of samples, photographic image processing, maintenance of lab equipment and supplies and the development/refinement of new sample preparation techniques as needed.
Qualifications: An advanced degree in Physics, Chemistry or Materials Science with proven TEM experience. 5 - 10 years of experience in TEM analysis and sample preparation and analysis of semiconductor materials and devices is desirable.
Hi All I have a need for a second hand Philips CM12 or CM20 preferably with compustage. If there is anyone with one to sell or is thinking of upgrading? I would take an instrument that is underused and give full access to the owner if that would help. The need is fairly urgent so a speedy reply would be appreciated even if only to express an interest Please reply directly to me and not the list. Many thanks
Chris
Chris Gilpin Biological Sciences Electron Microscope Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 161 275 5170 fax +44 161 275 5171 http://www.biomed.man.ac.uk/biology/emunit/emhome.html
} As was mentioned previously, the size of the skirt } is directly related to the gas pressure and one wants to operate as } low as possible for the samples being examined. I use 2.0 torr for my } polymer samples and lower for others. Also, higher accelerating } voltages reduce the skirt size but at an obvious cost if doing EDS of } light elements.
As people who know me will already know I am a biologist using ESEM. I almost always view hydrated samples. Try keeping a sample wet at 2.0 Torr! There are many people who use an ESEM to look at dry uncoated samples and in this case there are a number of parameters available for change. In any general discussion on the list remember that not all samples and imaging requirements are the same. Sounds like lots of discussion for the ESEM session at MSA and also the users group meeting.
Chris
Chris Gilpin Biological Sciences Electron Microscope Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 161 275 5170 fax +44 161 275 5171 http://www.biomed.man.ac.uk/biology/emunit/emhome.html
Hi, We have a large staff and many students. We have about 22 thousand dollars worth of various kinds of diamond knives. We always buy DIATOME. The quality is excellent and the service and support superior. I would not consider buying any other knife from another company. In 15 years of buying and using these knives, I have never been sent a poor one or one with any detectable flaw. I do not even "check" them out anymore when we get one resharpened. I know it is good. (I have no stock in EMS who sells these knives). Sincerely, Hildy
Kevin Mackenzie wrote: =============================================== I am trying to attatch various small samples (Diatoms, Microspores and Cecaria) onto Poly-L-lysine coated coverslips, but am finding that the samples do not attach.
Does anyone have any ideas? ================================================ So long as these are relatively "free flowing" powders, larger than about 4 um, then one of our Tacky Dot (TM) Slide products should work fine in this application. In addition, there is the added bonus that the particles are mounted in an orthogonal array, making it possible to do analytical work on the powder far more quickly if not also more accurately.
Information about Tacky Dot Slides can be found on our website.
Disclaimer: SPI is the sole worldwide manufacturer, under license from DuPont, of Tacky Dot Slides so we have a vested interest in seeing that more are used! We know of no other product like this one so there are no other references to be given.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Paiboon NUANNIN wrote: ================================================ We would like to examine a pore size of cellulose acetate with SEM. Please advise us, how to prepare the sample becuase it is very sensitive to vacuum. Can we dry by using a CPD? ================================================= Could you give more information about your system, for example, a) what is the cellulose acetate "wet" with?, b) what would be the expected pore size, and c) what is the physical form of the sample, is it a thin film coating on a substrate or is it more of a bulk sample? With regard to cellulose acetate, I don't think we have ever found porosity in that polymer system. But then again, maybe we did not look hard enough either.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I have a customer who will be using SEM/EDS to characterize a mixed oxide layer (Fe-, Ni-, Cr-, Mn-, Si-oxides, approximately 1 micrometer thick) furnace-grown from the superalloy substrate. While measuring the desired features will be successful at a comfortable rate in the lab, the ultimate goal is to develop a low-prep, rapid, reliable process that can be used on the plant floor by semi-skilled workers.
I checked around and it looks like x-ray thickness measurement tools may do the trick. Many fluorescence units are portable, easily used, etc. and are appropriate to our sample size (approximately 1 CM2)
My questions: Do any of you use industrial-duty x-ray fluorescence thickness gages for measuring oxide layers over metal substrates? If so, how accurate, repeatable, etc. Any caveats?
If you would prefer to respond directly, I can be reached at:
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 Phone: (508) 750-1717 E-mail: crossman-at-osi.sylvania.com
Our web sites: www.sylvania.com www.siemens.com --
well I now know why I am having difficulty finding the Amoeba since according to the book I have just bought, at least the classic Amoeba Proteus is rare in the wild!!! I remember being told by a retired microscopist that it was rare also. So at least I can take comfort in the fact that its more than likely there are none in my samples rather than my mis-identification! But I will not be giving up the search as everyone needs a mission in life *laugh*
Conrad
------------------------------------------------------------------------ ----------- "Any sufficiently advanced technology is indistinguishable from magic" ----------------------------------------------------------- Arthur C Clarke ----
I finally got a good book on Protozoa identification. In the end I found that the place I got my Microscope had a good book. Only 4.40 uk pounds! Thats the kind of price I like :)
Conrad
------------------------------------------------------------------------ ----------- "Any sufficiently advanced technology is indistinguishable from magic" ----------------------------------------------------------- Arthur C Clarke ----
Hello, I agree completely with the praise to DIATOME. Well deserved indeed. Sally
On Tue, 8 Jul 1997, HILDEGARD CROWLEY wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Hi, } We have a large staff and many students. We have about 22 thousand } dollars worth of various kinds of diamond knives. We always buy } DIATOME. The quality is excellent and the service and support superior. } I would not consider buying any other knife from another company. In 15 } years of buying and using these knives, I have never been sent a poor one } or one with any detectable flaw. I do not even "check" them out anymore } when we get one resharpened. I know it is good. (I have no stock in EMS } who sells these knives). } Sincerely, } Hildy }
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi YuHui, You can try using Cryo-SEM. Put a drop of beads in suspension onto a substrate, blot excess solution using filter paper, and fast-freeze it. In order to expose the surface for SEM viewing, you need to etch the ice on surface, but the bottom part of beads are still embedded in ice. Then apply cryo-coating and cryo-observation in a cryo-SEM. If you have any questions, please contact me offline.
Ya Chen
Ya Chen
========================================================================= \ / Integrated Microscopy Resource (IMR)-- \ / __ an NIH Biomedical Research Resource TEL : 608-263-8481 \/ / / University of Wisconsin-Madison FAX : 608-265-4076 / / / 1675 Observatory Drive #159 Email1:ychen14-at-facstaff.wisc.edu / /__/_ Madison, WI 53706 Email2:chen-at-calshp.cals.wisc.edu ========================================================================= IMR WWW Home Page: http://www.bocklabs.wisc.edu/imr.html
The Integrated Microscopy Resource and Carnegie Mellon University will be sponsoring a symposium and short course on multi-photon excitation imaging, August 9-10, 1997, in Cleveland Ohio.
We have never gotten a bad diamond knife from MicroStar in the 10 plus years we have been dealing with them. We have at least a dozen knives and get 2 or more resharpened every year.
} } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } At 12:16 PM 7/8/97 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America Scientific Director, ICBR Electron Microscopy Core Lab PO Box 118525 Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
We are trying to fix diatomaceous algae for SEM. Does anyone have any suggestions or warnings they could pass along.
Specifically, we are interested in the best solution for long-term storage (greater than 3 months).
The four suggested so far are: 1)lugol's solution, 2)formalin, 3)1% glutaraldehyde, 3)2% paraforaldehyde EM grade, and 4)a combination of 1% glutaraldehyde and .1% paraformaldehyde.
If you would like to post directly to me, I would be happy to submit a summary to the group.
Dear Microscopists, For the Microscopy and Microanalysis '98 meeting in Atlanta, the local arrangements committee is seeking a volunteer to organize a fun 5K run/walk for Sunday morning, July 12, 1998. For further information, please contact me off-line, at my e-mail address: yolande.berta-at-mse.gatech.edu Yolande Berta Georgia Tech School of Materials Science and Engineering 778 Atlantic Dr. Atlanta, GA 30332-0245 (404)894-2545
My two cents worth on EDS at elevated pressures, I published results of quantitative analysis of 70 microns Cr-spinel in Mg-olivine matrix at pressure in the specimen chamber from 1 Tr to 16.6 Tr with still good GSE resolution. Even at 10,000x magnification and analytical window width 4.5 microns, there is small but significant contribution from Mg-olivine towards Cr-spinel composition. And yes, changing working distance (shorter) and accelerating voltage (higher) can minimise skirt. My advice is if you have to do it in ESEM, go to the lowest possible pressure in the specimen chamber to avoid charging and try WD and kV for the best results. For biological specimen use a cold stage and keep a specimen at minimum temp. close to 0 deg. In this case you will still deal with a fully hydrated specimen at relatively low pressure of water vapor in the specimen chamber ( 4.647 Tr at 0.2 deg. C) Wis Jablonski OiC EM/X-ray Microanalysis Unit, CSL, Uni of Tasmania
Ref: W Jablonski, ESEM-2020-a key research tool in the university environment. The Third Biennial Symposium on SEM Imaging and Analysis: Applications and Techniques, Proceedings, Australian Microbeam Society, Feb 15-17, Melbourne 1995, pages16-17.
PS More recent and quantitative work by Bilde-Soerensen could be a good help.
Dear Garry, If you have done biological EM then metallurgical EM should be a breeze. Although my degree is in Microbiology, all my EM has been in Metallurgy and Materials Engineering. There is much more SEM than TEM and a lot of EDX, including the problems of quantitative EDX. Specimen preparation is straight forward and can consist of just putting the metal piece on a stub and into the SEM. You need to learn all about EDX and some basic principals of metallurgy. Electropolishing is just recipe following and there are some neat instruments, like ion beam thinners, to help you. It depends on what the particular lab you work at specializes in. I don't think there is anything as difficult as biological TEM specimen preparation and ultramicrotoming in the metallurgical field. The trick is to get the training you need from someone who knows it well.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
Further to my mail on EDS at elevated pressures: I would like to clarify the sentence No2: Even at 10,000x magnification and analytical window width 4.5 microns, there is a small but significant contribution from Mg-olivine towards Cr-spinel composition at the lowest ( in this case) 1 Tr specimen chamber pressure. Hope this will help. WJ
} Help! I need a source for indium foil. I need to use this to pick up } particles and residue from the equipment in our wafer fab.
Try ESPI, phone toll free (800) 638-2581, fax (818) 889-7098, at 5310 Derry Avenue, Agoura, CA 91301. Their catalog lists 9 different thicknesses each in 3 purity grades!!
I have no connection with them, I merely drool over their catalog occasionally.
Ritchie
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
I have to admit a very need solution. Being restricted on budget AND the exchange rate! (totally unfair) We are using a cheap solution. We produce envelopes from a ~ 5cm x 3cm piece of lint free paper (lens paper), staple it closed and CPD the normal way.
} For particulate samples that cannot be air dried, I sandwich the } sample between two Millipore filters held in a Millipore Swinney stainless } steel filter holder. It is designed to fit on a syringe, but I cut the } ends off (which allows for good fluid exchange), and run the holder through } the dehydration series and CPD. Alternatively, some samples can be dried } in HMDS or TMS, and you can skip the critical point dryer. } Also, I recomend using conductive carbon tabs for your adhesive. } They give a nice smooth background, and are more conductive than } double-stick tape (mine came from Pella, but other companies may also } offer them). } Please feel free to contact me off-line for more details if necessary. ## [########] ## ## ## ## ## Stephan H Coeztee Electron Microscope Unit Private Bag 3 Wits 2050 South Africa
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
G'day,
Do any of you have hints and advises to give me about studies of gels with TEM ? 1) What kind of preparation would you use ? 2) What would you characterise first ? 3) Would try to go for cryo TEM ? 4) Would you do replica or try to use a cold stage ?... Your experience is more than welcommed ! Thanks.
I have made a note in my palmtop to remember to bring the details of the protozoan book that I bought into work so I can send them to this list.
I can't say how good it is when compared to other possible books due to not having seen other books to compare it with! but I thought my little Observers Book was good value for 2 UK pounds and that book was concerned mainly with pond life in general. This book costs just over twice as much and is full of detailed pictures of various protozoan including of course several types of Amoeba which I am currently searching for...!
It is quite interesting though how different pictures in books can be with respect to certain creatures since I think the little observers book has a better drawing of COLEPS than this protozoan book in my opinion as it resembles a 'knurled barrel' whereas in this protozoan book it doesn't give so much an impression of a 'knurled' surface which is how it appeared to me under the 'scope!
I liked the warning about culturing Amoeba in that if you do it at too high a temperature you can favour the culture of a LETHAL PATHONOGENIC species of amoeba!!! although it says if careful it is unlikely as the temperatures it quotes are above 35C which is hot for a country like the UK!!! unless of course the central heating is turned up.....
For any amateurs on this list in the UK I obtained this book from Brunel Microscopes for the cost of 4.40 UK Pounds.
Conrad
------------------------------------------------------------------------ ----------- "Any sufficiently advanced technology is indistinguishable from magic" ----------------------------------------------------------- Arthur C Clarke ----
Dear all, we are in a desperate search for an old vacuum electronic module E-U12A Philips EM400 (Philips Cat. No. 532269514353) as our old one is down and we cannot afford to buy a new one from Philips - the old parts are outrageously expensive. Isn't there someone owning an old Philips 400 that is being replaced and could give us the module?
Thank you for any help or suggestion.
Pavel Hozak
__________________________
Pavel HOZAK, PhD Inst. of Experimental Medicine Dept. of Cell Ultrastructure & Molecular Biology Videnska 1083 142 20 Prague 4 - Krc Czech Republic
Laura L. Estok Asst. to the President M.E. Taylor Engineering, Inc. 21604 Gentry Lane Brookeville, MD 20833 Phone: 301-774-6246 * FAX: 301-774-6711 * e-mail: Metengr-at-aol.com
I'm a Metallugical engineer taking Ph.D. course on Biomaterials. My thesys is on titanium implants for odontology. My M.Sc. thesis was on a beta titanium alloy for aircraft industry and I did TEM characterization of this alloy. Now, I'm on the opposite way: I need to do histological cuts and analisys of the specimens (titanium implants inserted into the tibiae of rabbits).
I'd like you to give me some information about the interpretation of histological specimens on laser confocal microscopy or optical microscopy. First, about specimens preparation: We have a diamond wheel machine (ISOMET). I intend to embed the specimen with resin and gently cut (low speed, low weight). To obtain a histological slice, must I grind it? Till how many microns? Second: How can I quantitatively characterize de degree of osseointegration?
I hope you can help me.
Yours sincerely,
Marcelo Henrique Prado PEMM - COPPE/UFRJ Po.Box.:68505 Cidade Universitaria - Ilha do Fundao Rio de Janeiro-R.J. CEP.: 21941-900
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
REGARDING Indium foil
Beckey,
The best source is the Indium Corporation of America, 1-800-4-indium. I've used the foil to capture particles for years with excellent results. I work as a semiconductor failure analyst, 19 years here at Motorola.
Hope this is the information you need. Have a nice day!
Hi, I hope that the I didn't imply that the little book that I have bought had photographs in it! It has only drawings but still at 4.40 its good value I think. I intend to post the book details when I remember to bring them in!
Conrad
------------------------------------------------------------------------ ----------- "Any sufficiently advanced technology is indistinguishable from magic" ----------------------------------------------------------- Arthur C Clarke ----
Mary and Garry wrote about switch from Biological to Metallurgical EM:
Although I agree that sample prep can be far more demanding with biological specimens, as far as TEM is concerned, one thing biologists will most likely lack is training in crystallography. Most biologists I've met are not fond of reciprocal space. Garry, if you want to switch to crystalline solids, take a few courses in crystallography. Materials Science is far more than just generating a conventional TEM image.
Ciao for now, Ken
Kenneth JT Livi Department of Earth and Planetary Sciences 34th and Charles Streets Johns Hopkins University Baltimore, Maryland 21218 USA Phone: (410) 516-8342 Fax: (410) 516-7933 e-mail: klivi-at-jhu.edu
Can any of our German colleagues help with translation please? Came across the term PHOSPHORWOLFRAM acid in a technical paper. Suspect it is phosphoric acid but want to be sure before proceeding. Thanks in advance Ronnie Houston Texas Scottish Rite Hospital for Children Dallas
In message {33C3BC88.5577-at-airmail.net} writes: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Can any of our German colleagues help with translation please? Came } across the term PHOSPHORWOLFRAM acid in a technical paper. Suspect it is } phosphoric acid but want to be sure before proceeding. } Thanks in advance } Ronnie Houston } Texas Scottish Rite Hospital for Children } Dallas
Ronnie, wolfram in the old Germanic (I think) name for tungsten, so you have phosportungstic acid there, which when dissolved in water to a few percent is good ol' PTA, commonly used as a negative stain for viruses, bacteria, particulates in transmission electron microscopy.
--
Gib Ahlstrand, MMS Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
Plato: "When the mode of the music changes, the walls of the city will shake."
Chuck Berry: "There's a whole lotta shakin' goin' on!"
I have used spot modes to do EDX "maps" on the fly.
If I suspect a high concentration of a particular element in a specific local area of an image, I start acquiring and then move the spot onto and away from the feature of interest while simultaneously watching the growth of a peak for the element of interest. It is a crude but effective way of matching the location of particular to features observed.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} I travel the world teaching practical electron microscopy and it worries me } the emphasis that people place on "Spot Mode". I do not teach the use of } spot mode for the following reasons- } } 1. Just because the spot appears in a certain position on the screen } is this its correct position. I have found machines many microns out of } step in X and Y directions. } } 2. Specimens charge and switching out of spot mode does not give the } operator an easy opportunity to see if the spot had moved during the } analysis. } } 3. Spot mode gives a false sense of analytical volume, no matter how } small that spot may be we are almost certainly evaluating microns of } material. } } Do others worry about spot mode accuracy, do others test the spot mode } accuracy? Is it not better to simply increase the magnification watching } the area of interest all the way up before the analysis and all the way } down after the analysis? } } What do you think?
While the spot mode leaves a lot to be desired, particularly if the user isn't aware of the problems, analysis in raster mode is not without its difficulties. Most SEMs use some sort of of sychronisation of the line and frame scan signals. If so, the analysis obtained in raster mode is biased towards one edge and one corner of an area which may not even be identical with the image area - the beam 'waits' at the beginning of each line and each frame to synch, and the display may actually be blanked for a small distance after the scan starts and before the scan ends (over scanning) to hide image distortions arising from hysteresis, which is usually most evident at the edges of the scans.
Get a nice dirty specimen and scan it with a coarse raster until the contamination builds up. Now image the area at a lower mag, and look at the contamination pattern - there will probably be a heavier contamination line down one edge (the line scan 'wait') and a spot at one corner (the frame 'wait'). In addition, you may also be unlucky enough to find that the lines making up the coarse raster are bent at each end - you don't see the distortion in your images however because this part of the frame is blanked from display, but it will contribute to an EDX analysis.
More modern SEMs have reduced these problems, but I believe that they are still present in many.
I think that this demonstrates that whatever 'employers' might want, and manufacturers try to provide, for anything beyond the most basic EM, you need a skilled and trained operator with a full understanding of the instrumentation, and the time to fully check and calibrate the machine (who needs to be properly paid for undertaking a highly sophisticated and skilled job). Otherwise you get results that are at best doubtful and at worst wrong.
Ronnie Houston wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Can any of our German colleagues help with translation please? Came } across the term PHOSPHORWOLFRAM acid in a technical paper. Suspect it is } phosphoric acid but want to be sure before proceeding. } Thanks in advance } Ronnie Houston } Texas Scottish Rite Hospital for Children } DallasDear Ronnie,
This sounds like a "common" rather than proper chemical name. If it helps at all, Wolfram is the source for the chemical symbol "W", for tungsten. I would try looking up tungsten salts of phosphoric acid.
Good luck.
Barbara Foster Consortium President Microscopy/Marketing & Education 53 Eton Street Springfield, MA 01108 USA (413)736-6931 fax: (413)746-9311 email: mme-at-map.com (
Linda Barthel asked for a method to localize glycogen in cryosections. I sent her some background information needed in the microscopic study of glycogen, and I repeat them for all researchers interested in the subject.
The problem of glycogen is more complex than commonly appreciated, and the understanding of this complexity is a sine qua non condition in the microscopic study of glycogen.
Glycogen in the cell appears in the organelles, GLYCOSOMES, composed of glycogen and enzymes involved in glycogen synthesis and degradation. The structures stained by uranium and lead salts, and interpreted in EM as "particles of glycogen" represent in fact the protein component of glycosomes. Glycogen does not react with the ionic stains, but it can be demonstrated histochemically, by the PAS technique (periodic acid - Schiff reagent) in LM, and by the modification of this procedure (Thiery technique) in EM.
The problem is that glycosomes are easily destroyed during tissue processing. The most common destructive factor is the change in pH which breaks the bond between glycogen and protein. The effect is that the soluble protein component (enzymes) is washed out, and glycogen which is not fixed, floats in the cell and aggregates into clumps. The acidic treatment is inherent in the PAS procedure where periodic acid is used, therefore, in the vertically processed slides, the unfixed glycogen often accumulates as crescents at the bottom of the cells (the effect well known in the classical histochemistry).
In EM the common destructive factor is uranyl acetate (strongly acidic) used before tissue dehydration. In tissue processed without uranyl acetate glycosomes appear intact, even after the priodic acid treatment in Thiery technique, because the histochemical reaction is performed on sections which are already embedded in the resin. This embedding prevents the floating of the unfixed glycogen.
Freezing seems to be another destructive factor for glycosomes. Raether et al, 1977 (Z.Parasitenkunde, 54, 149) used deep-freezing of Entamoeba cultures, and found that only a few amoebae retained normal structure. Their micrographs indicate that in the destroyed organisms the glycosomes were destroyed.
Additional complication is that the described factors affect only free glycosomes, whereas others, which are bound to different cell structures remain resistant.
The review of glycosomes was published by K.K.Rybicka, 1996, Tissue & Cell 28 (3) 253-267.
} Can any of our German colleagues help with translation please? Came } across the term PHOSPHORWOLFRAM acid in a technical paper. Suspect it is } phosphoric acid but want to be sure before proceeding. } Thanks in advance } Ronnie Houston } Texas Scottish Rite Hospital for Children } Dallas
Ronnie,
"Wolfram" is the former chemical name for the element "Tungsten," hence the symbol of "W" on the periodic table. I think it's safe to assume that PHOSPHORWOLFRAM ACID is the same as PHOSPHOTUNGSTIC ACID. Phosphotungstic acid is listed in references as a stain for EM work, but I have no experience with it myself.
Jim Passmore Analytical Chemist Cryovac North America Duncan, SC
This may have nothing to do with your problem and others on this listserver may understand my story better than I do, but you may also want to note the batch of your collodion as well as the cytochrome c. We do routine TEM autoradiography which involves coating slides with a 1% collodion/amyl acetate solution, then placing the sections on the slides, carbon coating, processing the sections through autoradiography and placing grids on the sections, then removing the collodion before scanning on the TEM. At one point a couple years ago we ran into real problems removing the collodion after the procedure - the sections were removed before the collodion!! After much investigation and many trials a friend of mine finally spoke with the chemist working at the company we bought our stock collodion solution from. Apparently they had switched to a newer? better? safer? method of preparing the stock collodion. We now specially order collodion prepared somehow with ethanol and ethyl ether - and have never had a problem since!!!
Pat Hales McGill University Dept. of Anatomy & Cell Biology hales-at-hippo.medcor.mcgill.ca
Anyone knows anything about pyroxylin? I would like to know what it is exactly, and if it might be more stable than some other films for coating grids for the TEM. (Note that I'm looking at thick stuff - no thin sections here!)
I will be purchasing a TOPCON SEM soon and I am interested in studying clays. I am most interested in the preparation techniques that investigators have used to get good pictures of kaolinite, smectite, illite, etc.
I have been using an X-ray diffractometer w/clays for years but wonder about how to mount oriented/unoriented specimens on those stubs.
I'd agree with the assessment of Diatome knives as being excellent. We have 6 of them here, and they all seemed to be of excellent quality. Of course, I also like Dupont and DDK.
My thoughts,
Garry
} ---------- } From: Greg Erdos[SMTP:gwe-at-biotech.ufl.edu] } Sent: 8 July, 1997 12:55 } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Re: Good diamond knives } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Mounting the specimen may be as simple as placing it onto a stub using a conducting adhesive, waiting 20 minutes for it to dry, and then running your new microscope at a low ( {5kV) accelerating voltage. Keep the specimen as small as possible to limit contamination. Do not fall into t= he trap of believing that every specimen should be evaluated at 25kV. Vary the kV to see how the information you visualise changes with beam penetration, there is always one particular kV that will best present the=
specimen features. Obtain a simple Monte Carlo program so that you can calculate the depth from which the backscatter and x-ray information is coming. Changing Z and tilt will vary the backscatter contribution to yo= ur image and change the way the information is presented.
However if you wish to carry out EDX analysis in the microscope you will=
need up to 25kV to enable a full range spectrum evaluation. In this case=
if the material is non conducting you will need to coat it, preferably wi= th carbon. The simplest carbon coating systems are desk top and may either = be self contained or as an accessory for a sputter coater. In either case a=
carbon string system should be all that is required. The coating system i= s not the most important feature but the type of string you use will decide=
how easy it is to coat the specimen. We find the thick string like a boo= t lace is the best. If you new SEM is of the variable pressure variety th= is will help you reduce any problems of specimen charge
For both imaging atomic number contrast and to better visualise your specimen for EDX evaluation you need to use a backscattered electron detector. Varying the kV, viewing by backscatter differing depths of information, you will effectively be able to section the specimen, from 5= kV through to the microscopes maximum.. The Monte Carlo program may be used=
Diffraction pattern indexing can be difficult even if you know the general structure, i.e. orthorhombic, because of the different length sides of the unit cell (let alone angles in other structures). Then, just because you can find a set of reciprocal space planes at the right distances and angles from each other doesn't mean that the structure factor allows for that spot to be strong in reciprocal space. So any indexing software usually requires that you know positions of atoms in the unit cells of all the structures that would potentially fit the pattern that you are trying to index. EMS can be run on unix or vax machines. You must not only know the possible structures but also a range of camera length.
But,...I have found an alternate although cumbersome and limited method: I have generated and used successfully in Microsoft Excel a 3 sheet spreadsheet that notifies you of all planes that match the input specifications of the pattern to be indexed. You must also input the cell parameters and cell angles. This does not take into account any structure factors. And, it is limited by computer RAM. If you are interested, I can tell you more how to generate it yourself -- better that my old one.
When staining sections of Douglas-fir twigs, I use safranine/fast green. I would like to see a stain difference between early and late wood, but with this stain I can only tell the difference by the size of the vessels and the thickness of their walls. I would like a cleaner method. Any ideas?
Hello everyone! I'm sending this message to you in behalf of Ms. Louise Tayl or because I believe that her question might get more replies from this netw ork. Please send your replies to Ms Taylor directly at this address:
179LOU-at-chiron.wits.ac.za Thanks!
At 1:51 PM 97.7.9, Ms Louise Taylor wrote: } Hi there histonetters } } I have a query from an electron microscopist in our department. They } are having some problems doing IHC on their samples, particularly } with HMB45 (DAKO). } Are there any suggestions out there regarding optimum dilution of the } antibody or conversely another source that works well at EM level. I } realise that this is not necessarily a histonet query, but I would } appreciate whatever contacts, info etc I can get. } } Many Thanks } Louise Taylor
Laurent NORMAND wrote: ================================================ Do any of you have hints and advises to give me about studies of gels with TEM? 1) What kind of preparation would you use ? 2) What would you characterise first? 3) Would try to go for cryo TEM ? 4) Would you do replica or try to use a cold stage ?... ================================================= There might be as many philosophies of approach as there are people doing this kind of work. My first thought is always to determine what kind of features are you really trying to resolve in the gel structure. For example , certain greases (they act as gels because they have "thickeners") contain various metal stearates that form fibrillar type networks, and the differences between samples show up by just taking the gel, often times not even diluting it but just smearing it out on a glass slide to the thinnest possible film, and after RT drying, carrying out Pt/C shadowing and replication and looking at it that way. You see the metal stearate structures quite nicely and they usually, or at least some of them, get picked up with the replica so you can even do ED and DF studies. And the experimental procedure is very definitely not a complicated one, using just conventional equipment that just about everyone has at their fingertips in their laboratories.
On the other hand, if you have something like an aviation jet fuel kind of gel, and one is interested in seeing the network features of the polymeric additives (e.g. the ones that are responsible for the gelling of the system) , freeze fracture TEM is more appropriately indicated. If you are seeking to resolve "micropores", you might see them this way as well. If the gel, such as a so-called "hydro gel" type system is being looked at, being aqeuous based, some freeze etching should also be done.
If there is some kind of a non-organic type additive, that would have enough electron density in its own right, and the goal is to see its degree of dispersion in the gel, then cryo-TEM would be our preference. If the gel is aqueous based, and the presence of pores is what is to be resolved, then we have looked for ways to precipitate silver chloride into the pores to serve as a "decorator".
The conspicuous absence of mention of SEM was not accidental. We have almost never found SEM to be good enough to resolve the kinds of features people want to see when doing this kind of work.
It did not sound like you are asking about things like "gel" spots in polymer films which of course would require entirely different approaches, none requiring cryo, except maybe for diamond knife thin sectioning.
Disclaimer: We have no connection with any of these techniques except that we do offer them as a service for clients wanting to do this type of work.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Dear Hank, When I prepare clays to see the crystal structure, I usually suspend the clay in alcohol to the consistancy of milk and then put a drop on a SEM stub with a small glass cover slip on top, to get a very smooth surface. Dry, then gold coat. This will allow you to see the "books" that are characteristic of kaolinite. I use this method to look at the different crystal shapes of kaolinite, halloysite and illite for a lab. You wrote: } } I will be purchasing a TOPCON SEM soon and I am interested in studying } clays. } I am most interested in the preparation techniques that investigators have } used } to get good pictures of kaolinite, smectite, illite, etc. } } I have been using an X-ray diffractometer w/clays for years but wonder } about how } to mount oriented/unoriented specimens on those stubs. } } I'd appreciate any help or references. } } Hank Bart Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
as it might be of interest for some of you, I wish to point you to a new listserver at the medical faculty of the Westfalian Wilhelms-University. It discusses anatomical and related topics. However, it is announced for the english and for the german language. On the other hand it has just started and traffic is still low; therefore it might become common to discuss in English as Germans appear sometimes to be a bit offish :-)
To subscribe send an e-Mail to Majordomo-at-medweb.uni-muenster.de with the following body text: subscribe ANATOMIE-D {your-eMail-address}
Sorry if I have annoyed people but with all due respect Ray, I was asked to share the information about this book by fellow amateurs. I admit that I may have rambled on a bit. Although I have been on the net for a long time, I guess with respect to this list I am one of those 'dreaded newbies'. and thought it was like a newsgroup and forgot it was composed mainly of proffesionals.
I only posted because it would have taken too much time to reply to everyone and I had better end this here before this becomes another rambling post!
Regards,
Conrad
} -----Original Message----- } From: Ray Hicks [SMTP:rh208-at-cus.cam.ac.uk] } Sent: Wednesday, July 09, 1997 6:54 PM } To: Conrad Perfett } Subject: Re: protozoan book } } Conrad, } } It's good that you've found some information useful to you, but why not } wait until someone asks before you share it? You'll notice that there } isn't very much spontaneous posting to the list by anyone else, about } microscopy but especially about their research interests. Remember that } the common interest of the list is microscopy, not microbiology, histology } or metallurgy. If, for instance, someone has a microscopy related } metallurgical question they post it and get an answer, generally from } another metallurgist, you don't tend to get "I looked at a piece of } pearlite yesterday - interesting eh?" type comments. If you did you might } get twenty anecdotes a day from each of the members of the list, and it } would stop being so useful. } } How about lurking around until someone asks a question that's in your area } of expertise, and then helping them out? } } By the way, my school teachers used to provide amoebae for practical }
Dear all, we are in a desperate search for an old vacuum electronic module E-U12A Philips EM400 (Philips Cat. No. 532269514353) as our old one is down and we cannot afford to buy a new one from Philips - the old parts are outrageously expensive. Isn't there someone owning an old Philips 400 that is being replaced and could kindly give us the module or to suggest a solution?
Thank you for any help or suggestion.
Pavel Hozak
__________________________
Pavel HOZAK, PhD Inst. of Experimental Medicine Dept. of Cell Ultrastructure & Molecular Biology Videnska 1083 142 20 Prague 4 - Krc Czech Republic
I am working on single aerosol particle analysis using windowless SEM/EDX system. I hope to quantitatively analyze the contents of C, N, and O in microparticles. CITZAF program has been known to work for particles, so I am looking for where I could get it. I know it is a shareware and was distributed by Caltech. However, Dr. Armstrong seems not working over there any more. Would someone kindly help me to have a copy of CITZAF program? Many thanks in advance.
Sincerely yours, Chul-Un Ro
Chul-Un Ro, Ph.D Department of Chemistry University of Antwerp B-2610 Wilrijk Belgium
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Howdy, We have a JEOL 2010 STEM that must be isolated from a huge vibrating air compressor in a different part of the building (no they won't move it or isolate the compressor). Our plan is to cut the floor (8" of high density reinforced concrete sitting on dirt) around the microscope. I am interested in tips and lessons learned,e.g., width of cut, filler material in the cut, is it ok to tile over the gap, how far from the scope to place the cut, etc.
Brad Storey Materials Scientist Argonne National Lab - West P.O. Box 2528 Idaho Falls, ID 83403 Ph. 208-533-7685 Fax 208-533-7683 e-mail brad.storey-at-anl.gov
Greetings, Any of you erudite types out there know the origins of the word "Wolfram"? or why or when the word "tungsten" (which I believe comes from sweedish words for "heavy + steel") came to be the prefered word? Please forgive my curiousity if this is too far off the microscopical axis for your taste.
Short article on pyroxylin in Merck Index. Cellulose nitrate in old film base and plastics is not stable over decades, but this probably does not matter for your needs. Note hazards.
I am currently staining milk proteins with phosphotungtic acid (2%/water ph 7.3) and uranyl acetate (1%/water) and often I do get what I called a background consisting of very small (about 20-40nm) bubbles-like structures that interfere with my samples...
I also recall seeing the same thing with viruses but this time it is rather annoying because it also looks a lot like my protein...
Does somebody out there has a explanation to this phenomenon....I though maybe it was perhaps a hydrophobic reaction between the formvar and the stain or irregularities/defects in the formvar film....
thank you,
Diane Montpetit Food research center agriculture canada st-hyacinthe, quebec, Canada fax 514 773 8461 tel; 514 773 1105 e mail; montpetitd-at-em.agr.ca
The tungsten mineral wolframite was known in the tin mines of the Saxony-Bohemia region long before the element itself was discovered. In 1781, the Swedish chemist Scheele, who had been working with the stony mineral, elucidated the composition of this mineral to be a compound of calcium with an unknown acid. The acid-forming element thus discovered was named tungsten by A. F. Cronstedt in 1755. He derived this name, from the Swedish words "tung", meaning heavy, and "sten", meaning stone.
Wolfram
In 1783, the brothers J.J. and F. de Elhujar found that wolframite also contained tungsten. After success in obtaining metallic tungsten from wolframite, and gave it the name "wolfram" The origin of the word is not clear, but it is assumed to be derived from German words "Wolf" and "Rahm" or Swedish word "wolfrig".
Wolfram is the official international alternate name for tungsten. Tungsten is preferred in the United States.
Hello fellow microscopists, I'm in the process of using the DGD resinless embedding technique developed by Capco, Krochmalnic and Penman and I'm looking for: 1)Short cuts-the ethanol, nbutanol to DGD transition is very long (6 hours) and I am working with monolayers, so is there a poblem with this version: Graded ethanol dehydration to 100% 30 min 100% ethanol 4 changes 1 hr 1:1 nbutanol:100% ethanol 30 min 100% nbutanol 1 hr 1:1 nbutanol:DGD 45 min 100% DGD (with DMSO) 1 hr RT harden O/N
2) Also any suggesstions on how to harden this wax on tissue culture dishes, coverslips (glass) and platic slides will be greatly appreciated. Thank you.
Dear all, we are in a desperate search for an old vacuum electronic module E-U12A Philips EM400 (Philips Cat. No. 532269514353) as our old one is down and we cannot afford to buy a new one from Philips - the old parts are outrageously expensive. Isn't there someone owning an old Philips 400 that is being replaced and could give us the module?
Thank you for any help or suggestion.
Pavel Hozak
__________________________
Pavel HOZAK, PhD Inst. of Experimental Medicine Dept. of Cell Ultrastructure & Molecular Biology Videnska 1083 142 20 Prague 4 - Krc Czech Republic
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi Brad, I have done a bit of work staining core samples and twig sections of Doug fir in an attempt to age the wood, and safranin staining was the best way to evaluate this. Unfortunately, the only difference between early and late wood is vessel diameter (ie. secondary cell wall thickness). There may be differences in minor cell wall components from early to late wood but none of them (I predict) would show as good a demarcation as safranin does in staining lignin. Don't overstain with fast green as you can lose contrast, in fact, it can be left out altogether because you are not interested in staining cytoplasmic components. Remember that in Doug fir there will be a gradual transition from early to late wood. You have to estimate where the change is based on an arbitrary cell wall thickness that you get to choose (I get a real rush making executive decisions like that). Cheers, John
================= C. John Runions, Ph.D Section of Ecology and Systematics Corson Hall Cornell University Ithaca, New York USA 14853
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi Brad, I have done a bit of work staining core samples and twig sections of Doug fir in an attempt to age the wood, and safranin staining was the best way to evaluate this. Unfortunately, the only difference between early and late wood is vessel diameter (ie. secondary cell wall thickness). There may be differences in minor cell wall components from early to late wood but none of them (I predict) would show as good a demarcation as safranin does in staining lignin. Don't overstain with fast green as you can lose contrast, in fact, it can be left out altogether because you are not interested in staining cytoplasmic components. Remember that in Doug fir there will be a gradual transition from early to late wood. You have to estimate where the change is based on an arbitrary cell wall thickness that you get to choose (I get a real rush making executive decisions like that). Cheers, John
================= C. John Runions, Ph.D Section of Ecology and Systematics Corson Hall Cornell University Ithaca, New York USA 14853
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi Brad, I have done a bit of work staining core samples and twig sections of Doug fir in an attempt to age the wood, and safranin staining was the best way to evaluate this. Unfortunately, the only difference between early and late wood is vessel diameter (ie. secondary cell wall thickness). There may be differences in minor cell wall components from early to late wood but none of them (I predict) would show as good a demarcation as safranin does in staining lignin. Don't overstain with fast green as you can lose contrast, in fact, it can be left out altogether because you are not interested in staining cytoplasmic components. Remember that in Doug fir there will be a gradual transition from early to late wood. You have to estimate where the change is based on an arbitrary cell wall thickness that you get to choose (I get a real rush making executive decisions like that). Cheers, John
================= C. John Runions Section of Ecology and Systematics Corson Hall Cornell University Ithaca, New York USA 14853
In message {s3c4b19e.001-at-EM.AGR.CA} Diane Montpetit writes: } hello everyone, } } I am currently staining milk proteins with phosphotungtic acid (2%/water } ph 7.3) and uranyl acetate (1%/water) and often I do get what I called a } background consisting of very small (about 20-40nm) bubbles-like } structures that interfere with my samples... } } I also recall seeing the same thing with viruses but this time it is rather } annoying because it also looks a lot like my protein... } } Does somebody out there has a explanation to this phenomenon....I though } maybe it was perhaps a hydrophobic reaction between the formvar and } the stain or irregularities/defects in the formvar film.... } } thank you, } } Diane Montpetit } Food research center } agriculture canada } st-hyacinthe, quebec, Canada } fax 514 773 8461 } tel; 514 773 1105 } e mail; montpetitd-at-em.agr.ca
Perhaps you could modify your formvar/grid preparation techniques to see if it makes any improvement. You could try: 1. Coat the Formvar coated grids with a thin layer of evaporated carbon. 2. Treat the Formvar coated grids in a glow discharge device if one is available. Either treatment may improve spreading and negative staining of your samples.
I looked at milk proteins, too, a few years ago using PTA staining and I don't recall any problems. The "bubbles" you see in the background may be due to contaminant that got on the Formvar film during preparation or handling, so look at your Formvar coating technique to see if you can clean it up; use pure double distilled water to float film off a previously cleaned glass slide, water in clean trough, etc, etc.
Good Luck!
--
Gib Ahlstrand, MMS Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
Plato: "When the mode of the music changes, the walls of the city will shake."
Chuck Berry: "There's a whole lotta shakin' goin' on!"
} Date: Wed, 9 Jul 1997 16:22:22 -0400 (EDT) } From: Leclerc Jean {leclercj-at-magellan.umontreal.ca} } To: Microscopy Association of America {Microscopy-at-Sparc5.Microscopy.Com} } Subject: Pyroxylin coating
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hi there!^ } } Anyone knows anything about pyroxylin? I would like to know what it is } exactly, and if it might be more stable than some other films for coating } grids for the TEM. (Note that I'm looking at thick stuff - no thin } sections here!) } } Thanks! } Our lab uses Pyroxylin, also known as parlodian or collodion, frequently, as it is easy to use. We buy as a 2% solution in amyl acetate from an EM supplier. The method: one drop on DDW in a 10mm diameter dish, let the film dry and then remove with the tip of a glass pipet to remove any dust particles, followed by another drop and let dry (interference colors become visible when viewed at a low angle). Then very gently place clean 10 to 25 grids dull side down on the film's surface.Carefully lay a piece of parafilm on the surface, pull the parafilm back up, and the grids plus film come along. Place in petri dish to thoroughly dry, followed by carbon evaporation. It is a very quick method and suitable for a number of applications.
you didn't mention what sort of coating you are using on your grid. Could these bubble structures be defects in the plastic coating caused by moisture absorbed into the liquid plastic stock solution? I must admit they sound very small but you can get small semi-perforate holes when you try to make holey plastic films.
If they are present before sample and stain they would be more difficult to see but should show up with a small objective aperture and lower KV.
Sorry if you've already thought of this.
Malcolm Haswell e.m. unit University of Sunderland UK ----------
hello everyone,
I am currently staining milk proteins with phosphotungtic acid (2%/water ph 7.3) and uranyl acetate (1%/water) and often I do get what I called a background consisting of very small (about 20-40nm) bubbles-like structures that interfere with my samples...
I also recall seeing the same thing with viruses but this time it is rather annoying because it also looks a lot like my protein...
Does somebody out there has a explanation to this phenomenon....I though maybe it was perhaps a hydrophobic reaction between the formvar and the stain or irregularities/defects in the formvar film....
thank you,
Diane Montpetit Food research center agriculture canada st-hyacinthe, quebec, Canada fax 514 773 8461 tel; 514 773 1105 e mail; montpetitd-at-em.agr.ca
We have an older, DOS-based version of Image Pro Plus image analysis software that has us stymied. We are capturing video images of chloroplast movements and dumping them into a DOS-based computer. We want to be able to sum all of the white areas in a field and then express that total white area as a percentage of the total image area. It seems simple enough, but it's not. We can identify all of the "white" objects, we can define what we are calling 'white', we can get the area of each individual white object, but we cannot sum the areas of all of the white objects. Image Pro Plus corporation is no help. They sold the rights to the DOS version to another company and no one a IPP knows anything about it. They are quite willing to sell us the Windows version for $3500 but cannot help us with their older version. Does anyone out there in microscope land have experience with this program?
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Greetings, Any of you erudite types out there know the origins of the word "Wolfram"? or why or when the word "tungsten" (which I believe comes from sweedish words for "heavy + steel") came to be the prefered word? Please forgive my curiousity if this is too far off the microscopical axis for your taste.
Back in my early graduate school days when I used to be a botanist, we used a stain called phloroglucinol which gave excellent demarcation of wood based on lignin content. I remember that you have to be very careful not to overstain, as everything will become crimson. The rest of the tissue, parenchyma etc. was contrasted with fast green. The stain can be modified to distinguish between deciduous and conifer wood (excluding Ginko), although I never tried this. The technique is described in Johansen's "Plant Microtechnique". I'm not sure if this stain would apply to your case, but it might be worth a try.
-=W.L. Steffens=- Department of Veterinary Pathology College of Veterinary Medicine University of Georgia Athens, GA 30602 USA http://www.vet.uga.edu/vpp/wls/steffens.html
The "bubbles" on the substrate that you describe are wetting artifacts...at least that is what I was told by an oldtimer. I get them consistently on negative stained viral samples, unless I incorporated a small amount (a very small amount) of bacitracin into the negative stain. I suppose some other surfactants would work, but bacitracin does the trick for me every time.
-=W.L. Steffens=- Department of Veterinary Pathology College of Veterinary Medicine University of Georgia Athens, GA 30602 USA http://www.vet.uga.edu/vpp/wls/steffens.html
My experience is better with isopropyl alcohol, more dilute suspension plus in some stubborn cases a litle ultrasonic bathing... Everything will be visible! Kris
} When I prepare clays to see the crystal structure, I usually suspend the } clay in alcohol to the consistancy of milk and then put a drop on a SEM stub } with a small glass cover slip on top, to get a very smooth surface. Dry, } then gold coat. This will allow you to see the "books" that are } characteristic of kaolinite. I use this method to look at the different } crystal shapes of kaolinite, halloysite and illite for a lab.
Disclaimer first: I'm not a chemistry/physics person, so please excuse me if this question is incredibly stupid :) Is it possible for a pigment to scatter/reflect light strongly enough to look like fluorescence? If so, how could you distinguish such behaviour from "real" fluorescence? I'm trying to work with some pigmented plant samples that "glow" under my rhodamine/Tx Red excitation/emission filters (standard fluorescence scope); the pigmented areas look normal under FITC and UV conditions. The pigment itself is red under regular light. We'd like to know if the pigment autofluoresces or if this is just some light/pigment interaction. I'm lost and our library isn't heavy on this type of references. Any of you "hard science" types willing to try to educate me?!
On Thu, 10 Jul 1997 wise-at-vaxa.cis.uwosh.edu wrote: } We have an older, DOS-based version of Image Pro Plus image } analysis software that has us stymied. We are capturing video images of } chloroplast movements and dumping them into a DOS-based computer. We want } to be able to sum all of the white areas in a field and then express that } total white area as a percentage of the total image area. It seems simple } enough, but it's not. We can identify all of the "white" objects, we can } define what we are calling 'white', we can get the area of each individual } white object, but we cannot sum the areas of all of the white objects. } Image Pro Plus corporation is no help. They sold the rights to the DOS } version to another company and no one a IPP knows anything about it. They } are quite willing to sell us the Windows version for $3500 but cannot help } us with their older version. } Does anyone out there in microscope land have experience with this } program?
Yes, a bit. Is the remaining non-white area contiguous? If so, measure that as the reciprocal of the white areas.
I'm trying to find a vendor who might sell a dissecting microscope that has 2 heads. We need something like this for demonstration purposes. It's a lot easier to use one of those than to keep switching chairs and asking if they see what we're talking about. So if anybody knows please send me the information so I can check into it further.
Thanks!
Paula = )
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
} I am currently staining milk proteins with phosphotungtic acid (2%/water } ph 7.3) and uranyl acetate (1%/water) and often I do get what I called a } background consisting of very small (about 20-40nm) bubbles-like } structures that interfere with my samples... } } I also recall seeing the same thing with viruses but this time it is rather } annoying because it also looks a lot like my protein...
This annoying phenomenon, sometimes called "champagning", may be caused by several factors. In my own experience, I would see it whenever using PTA in preparations with high protein concentrations. If you are quick, you can actually see the phenomenon taking place as the beam vaporizes the PTA. I believe that the proteins serve to sequester water which is then enrobed in PTA. When the beam strikes the hydrated proteins, the water is vaporized and bursts through the PTA crust (like erupting gases in magma) leaving behind uniformly sized, spherical, electron light areas that may be mistaken for viruses or protein subunits. One workaround may be to dilute the protein, use phosphomolybdic or silicotungstic acids as the negative stain and to dry the grids in a 60 degree oven over the weekend.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
I am looking for a software package to construct and visualize an amorphous 3-D SiO2 network with options that allow for insertion of other elements such as nitrogen, carbon, and hydrogen and also the possibility of forming various molecular groups such as silanol, siloxane, etc..
Does anyone know if such a software package exists?
} I am looking for an electron diffraction analysis software package. I have } to identify some particles (wether they're cubic, tetragonal, etc) and } there must be an easier way than to hand draw the expected diffraction } patterns....Does anybody know of something useful on the Web?
Try the EMS On-line soft running at WWW URL http://cimewww.epfl.ch/. This is a short version of P. Stadelmann EMS programme.
Sara, You can build a library of possible structures (lattice parameters, space group and if known the atom position in the cell to account for kinematical extinctions). Then you enter two or three diffracted vectors (lengths on the pattern, angles, approximate camera length) and the programme tells you on which zone axis of which structure you took the diffraction pattern plus the exact camera length needed to do the fit. If there are some ambiguities, you can also view/print the computed pattern=8A and more!
Philippe-A. Buffat
__________________________________________________________________ Philippe Buffat Ecole Polytechnique Federale de Lausanne (EPFL) Centre Interdepartemental de Microscopie Electronique Address: EPFL-CIME, Batiment MX-C, CH-1015 Lausanne, Switzerland Phone: +41(21)693 29 83 Fax: +41(21)693 44 01 (Central European Time) E-mail: philippe.buffat-at-cime.uhd.epfl.ch, WWW URL http://cimewww.epfl.ch/ ______________________________ Eudora F2.1 ___________________________
At 08:49 10/07/1997 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Wolfram is a German word and it's a reference to the gray color of "Tungsten" similar to the color of a wolf. Tungsten is a swedish word and means heavy stone. We use the word "Tunstene" in France, Wolfram is an alternate word but not very common. Salutations. ========================================================== Jacky Larnould mailto:larnould-at-mnet.fr voice:33 (0)4 67 72 28 26 fax :33 (0)4 67 79 54 90
I came across some sort of porous superlattice silicon(I think) in my rearch, I would be happy if someone provides me some information on diffraction patterns of porous superlattice silicon.
Thanks in advance
S Suder
Suli Suder Joule Physics Lab University of Salford Salford M5 4WT United Kingdom Tel: 0161 745 5000 ext. 53264 Fax: 0161 745 5119 E-mail: s.suder-at-eee.salford.ac.uk
Mark et.al. - We have the confocal listserver linked in our comprehensive listing of microscopy sites. When you go to it you will get this notice:
"Season's Greetings Our server is down for repairing. We will gradually bring the web up in the coming week. Please visit us after the New Year. Have a happy holiday!"
I think they are a bit premature! Seasons greetings Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 400+ Links, MSDS ************************ http://www.proscitech.com.au
_________________________ } Dear all, } Could someone please give me the address of the listserver for } confocal microscopy. } } Thanks a lot, } } Mark Munro } SAC Aberdeen } m.munro-at-ab.sac.ac.uk
Mark et.al. - We have the confocal listserver linked in our comprehensive listing of microscopy sites. When you go to it you will get this notice:
"Season's Greetings Our server is down for repairing. We will gradually bring the web up in the coming week. Please visit us after the New Year. Have a happy holiday!"
I think they are a bit premature! Seasons greetings Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 400+ Links, MSDS ************************ http://www.proscitech.com.au
_________________________ } Dear all, } Could someone please give me the address of the listserver for } confocal microscopy. } } Thanks a lot, } } Mark Munro } SAC Aberdeen } m.munro-at-ab.sac.ac.uk
Jean and all: Here is a copy from our on-line catalogue:
"PARLODION Pyroxylin, in strips. A highly purified form of cellulose nitrate; prepared for embedding tissu= es for sectioning and for making support films for EM grids from 1% parlodio= n in Amyl Acetate."
I prefer parlodion over formvar because it seems to release better when cast on a microscope slide, also is seems more suitable for thicker film= s. It comes in hard strips. Cut and weigh a suitable piece and then calcula= te the solvent required to make the percentage solution. Because it is very flammable parlodion it is frequently stored under water. Blot, then dry = in an incubator before weighing the small piece. Parlodion, like formvar requires=20 a carbon coating for stability in TEM. Butvar does not.=20 ProSciTech and I trust all other EM suppliers handle Parlodion. =20 Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 400+ Links, MSDS =20 ************************ http://www.proscitech.com.au =20 ---------- } From: Leclerc Jean {leclercj-at-magellan.umontreal.ca} } Hi there!=88 } =20 } Anyone knows anything about pyroxylin? I would like to know what it is } exactly, and if it might be more stable than some other films for coati= ng } grids for the TEM. (Note that I'm looking at thick stuff - no thin } sections here!) } =20 } Thanks! } ---------- } =20
At 10:49 AM 7/10/97 -0400, Michael Delannoy wrote: I'm in the process of using the DGD resinless embedding technique developed by } Capco, Krochmalnic and Penman and I'm looking for: 1)Short cuts- } the ethanol, nbutanol to DGD transition is very long (6 hours) and I am working } with monolayers
{snip}
It is often possible to dramatically speed up the dehydration steps of TEM specimen preparation by carrying them out in a laboratory microwave. The technique is described on pp352-353 of _The Microwave Cookbook for Microscopists._
disclaimer: Energy Beam Sciences manufactures laboratory microwaves, and has a vested interest in seeing more people utilizing this technique.
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
O.K. you professionals here's a real puzzler for you:
I have a user here who has about 24 SPI Slide-A Grid Boxes. They were a little dusty (they may have been used before - heavens not to say they didn't come absolutely pristine from SPI), and so he throughly washed them and rinsed them with Ethanol, only now he can not get them back open any longer! They are really stuck.
I don't know if he removed some type of lubication on the box, maybe the "natural" lube from the manufacturing process, or if he caused the plastic to swell with the EtOH, but I could certianly use any suggestion. Maybe we should throw this up to the group - surely someone has cleaned grib boxes before.
We can get the box lids to move upto 5mm or so but then they stop and with work we can get them to move back, but we still have freed any lids. We've tried prying up the lids a little from the back edges (where they slide along rasied ridges) - didn't help much. We've even tried soaking them again in water or EtOH hoping to provide some "lubrication" (there are no grids in the boxes, but didn't really want to soak them in some thing oily) - still with no success.
I oringinally asked this of Charles Garber (of SPI) and neither of us have come up with any ideas, but he did throw in the following:
Of the box parts:
} The plastic is an antistatic formulation however, alcohol being so } polar is not going to swell the base piece at least. If the alcohol } was hot or if it was in contact with the plastic for some extended } period of time, then who knows, but I just can not believe that the } alcohol would have done anything to it in the way you are } suggesting. } } The sliding top might be another story. I forgot the plastic that is } used for it, but it might be a clear PVC. But even then, room } temperature alcohol should not have had time to do anything. }
Any suggestions?
The things fools will do ....
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513-529-5712 Fax: 513-529-4243 E-mail: edelmare-at-muohio.edu
"640K ought to be enough for anybody." -- Bill Gates, 1981
ebs-at-ebsciences.com wrote: } It is often possible to dramatically speed up the dehydration steps of TEM } specimen preparation by carrying them out in a laboratory microwave. The } technique is described on pp352-353 of _The Microwave Cookbook for } Microscopists._
Another way to speed up dehydration is to do it chemically - use acidified 2,2 dimethoxypropane (DMP). For a monlayer complete dehydration will occur in a few min. I usually dehydrate small blocks (up to 3x3x3 mm) of plant tissue for 15 min, and then go directly to embedding medium:dimethoxypropane 1:1. Any residual water from the dehydration step will be removed during the first embedding step. However, I do not know if DGD can be mixed or react with DMP, so you have to test it first.
Any reason why you wouldn't use an anti-vibration platform? We have a Micro-g isolation table made by Technical Manufacturing Corporation for our JEM 1010. We were in a room next to the furnace and air conditioners for the whole building and NEVER had any problems. If you are interested, I have the information, so should JEOL, since they both have offices in Peabody, Massachusetts.
Cheri Owen Detroit Neurotrauma Institute Wayne State University Detroit
Microscopy & Microanalysis '97 in Cleveland is less than a month away! The deadline for early registration is this coming Tuesday, July 15. If you have not yet done so, register now to qualify for the lower rates. If you need registration forms, please call the business office at (800) 538-3672 . If you have internet access to the www, forms are available for downloading at http://www.bright.net/~strecker/msno/mm97.html.
If you are planning on attending and have not yet reserved hotel space, you must contact the hotels directly. Hotel information is also available from our web site or from the business office. I can also e-mail you this information if required. At our last planning meeting it was announced that there are no rooms available downtown Cleveland for Saturday, August 9, so if you need a room, please act now.
A special note to those who have been following the Mars Pathfinder program. A debate on the pros and cons of life on Mars will take place on Monday, August 11. Following this will be a presentation by Al Worden, Apollo 15 astronaut. Mr. Worden's talk will feature many of his breath-taking photographs from space.
---------------------------------------------------------------- Dave Strecker mailto:dave.strecker-at-ab.com Rockwell Automation/Allen-Bradley Phone: (216)646-3250 Component Engineering ND246 Fax: (216)646-3416 1 Allen-Bradley Dr. Mayfield Hts., Ohio 44124 USA ----------------------------------------------------------------
Sorry Geoff! I guess I was forgetting that other people may want to know the details of this book, so I am posting the details here:
A beginners guide to the collection, isolation, cultivation and identification of Freshwater Protozoa
Pblished in 1988 by Culture Collection of Algae Associaction (CCAP)
Freshwater Biological Association The Ferry House Ambleside Cumbria United Kingdom
ISBN 1 871105 03 X
but remember it has drawings not photos! still for 4.40 uk pounds you can't complain :)
} -----Original Message----- } From: Geoff McAuliffe [SMTP:mcauliff-at-UMDNJ.EDU] } Sent: Friday, July 11, 1997 4:34 PM } To: Conrad Perfett } Subject: protozoa book?? } } Dear Conrad: } } You have been on the MSA list praising a book on microscopy of protozoa } quite often. So what is the title of the book??????? I keep reading your } posts hoping to find some mention of the title, author, publisher, } something! Either post the title or email it to me, please. } } 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 e-mail: mcauliff-at-umdnj.edu } ***************************************************************
Greetings, I received many replies to my original query, and in case others are interested, here is the gist of them. Thanks to all of you who answered.
Hi Tamara,
If you have a proper filter setup, the emission filter should be excluding all the exitation wavelength. Therefore I think you are looking at autofluorescence. Any light that has not changed wavelength should be blocked.
Bob
On Thu, 10 Jul 1997, Tamara Howard wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Disclaimer first: I'm not a chemistry/physics person, so please } excuse me if this question is incredibly stupid :) } Is it possible for a pigment to scatter/reflect light strongly } enough to look like fluorescence? If so, how could you distinguish such } behaviour from "real" fluorescence? I'm trying to work with some pigmented } plant samples that "glow" under my rhodamine/Tx Red excitation/emission } filters (standard fluorescence scope); the pigmented areas look normal } under FITC and UV conditions. The pigment itself is red under regular } light. We'd like to know if the pigment autofluoresces or if this is just } some light/pigment interaction. I'm lost and our library isn't heavy on } this type of references. Any of you "hard science" types willing to try to } educate me?! } } TIA! } } Tamara Howard } CSHL } howard-at-cshl.org } } } }
Does anyone know where I can get a used LogEtronics 55-EM enlarger (or something similar with auto-dodge & burn-in capabilities)in good working condition?
Thanks,
Michael Coviello Lab Manager The University of Texas -at- Arlington
Hello again, DGD stands for diethylene glycol disterarate, which is a waxy "solid" at room temp (although sweats/melts at r.t. like the London resins-white,gold) and liquid at 70 deg.C. The references are: Pennman (1995) PNAS. 92:5251-5257 review Capco etal (1984) JCB. 98:1878-1885 These will get you started. Right now I've run some samples on tissue culture plates, coverslips and a glass slide (which got trashed). Separating the DGD from the plates was easy (light hammer tapping) but some cells did remain on the dish. The coverslips separated easily, so I've mounted a few and will be cutting soon (I can't see the cells so I'm sectioning blindly). One problem is the block sweating at the cell surface, I've placed everything in the fridge hopefully to stop this-cutting should be interesting. The second paper used glass petris which I don't have, but may have to get. The n-butanol did not dissolve the culture plate. I'll keep you posted, more suggesstions are welcome.
Sweating in Baltimore, Mike D.
P.S. I got my DGD from Polysciences who also sent a protocol. This stuff may work better with tissue blocks and pellets, monolayers are tricky.
An excellent source for used photographic equipment of all kinds, including digital imaging, is a monthly magazine called "Shutterbug". I think it's put out by the same people who do the "Computer Shopper" magazine. They began as an advertising magazine dedicated to used photo stuff, but have expanded into a large format glossy, general photography magazine. They still have loads of ads and classifieds for all sorts of used you-name-it relating to photography. It can be found in many camera shops and magazine newstands.
Randy Tindall Center for Electron Microscopy Southern Illinois University at Carbondale
Hi, In 25 years of being in a microscopy laboratory I have witnessed impossibly sticky grid boxes 3 times. Twice they were manufactured by LKB, once they came from another source. Each time the grid boxes were washed in alcohol. No time were we able to rescue the boxes. We threw them away. We now strictly wash all grid boxes with soap and water only. So sorry, HHC
In Image-Pro for DOS, after you count and measure all the white objects, the total area of these objects can be found by going to View-Statistics. The Sum measurement is the total area of all counted objects.
Additionally, to clarify a few points, Image-Pro for DOS was not sold to anyone - Media Cybernetics - the original developer of the Image-Pro family of products - still owns all rights to these products. However, the DOS versions of Image-Pro were discontinued quite some time ago (nearly 4 years). Although we make our best effort to support all of our products and customers, there have been at least 6 new Windows versions published since the last DOS version, and the vast majority of our customers have taken advantage of the upgrades or extended maintenance plans in order to stay 'current'. If you have registered your product with us, then you have probably seen the announcements and upgrade offers over the years.
If you, or any other DOS users out there, would like to upgrade to the latest version - Image-Pro Plus 3.0, please feel free to contact me directly.
-------------------------------------- Scott D. Ireland Regional Sales Manager Eastern North America & Latin America Media Cybernetics, LP "The Imaging Experts" Tel: (716) 473-0222 Fax: (716) 473-8048 scott-at-mediacy.com http://www.mediacy.com ---------------------------------------
---------- } From: wise-at-vaxa.cis.uwosh.edu } To: Microscopy-at-sparc5.microscopy.com } Subject: Image Pro Plus } Date: Thursday, July 10, 1997 9:21 AM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } To all, } } We have an older, DOS-based version of Image Pro Plus image } analysis software that has us stymied. We are capturing video images of } chloroplast movements and dumping them into a DOS-based computer. We want } to be able to sum all of the white areas in a field and then express that } total white area as a percentage of the total image area. It seems simple } enough, but it's not. We can identify all of the "white" objects, we can } define what we are calling 'white', we can get the area of each individual } white object, but we cannot sum the areas of all of the white objects. } Image Pro Plus corporation is no help. They sold the rights to the DOS } version to another company and no one a IPP knows anything about it. They } are quite willing to sell us the Windows version for $3500 but cannot help } us with their older version. } Does anyone out there in microscope land have experience with this } program? } } Yours in computer frustration, } } Bob } }
} } Does anyone know a TEM protocol for "Tungsten Shadowing" say, for DNA } strand enhancement? } } Thanks! } } Cheri Owen } Detroit Neurotrauma Institute } Wayne State University } Detroit
Cheri, There are two sections on tungsten shadowing in Chap 7 " High Resolution Shadowing" by Henry S. Slayter in the book: Electron Microscopy in Biology, J.R. Harris, ed. 1991, IRL Press at Oxford Univ. Press. ISBN 0-19-963215-4 (pbk)
cheers ed
Edward J. Basgall, PhD The Pennsylvania State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618
Often in our lab we are in a big rush to get prints right after the user has finished using the EM. I've sometimes just washed the negatives (3.25X4" Kodak EM film) just 2 minutes instead of 20, and then after I've printed them all, I go back and wash them for 30 minutes to remove any trace fixative that might still be remaining in the emulsion. This way I can shave 18 minutes off of the delivery time. I also print them with just a bit of water at their edges so that they are not completely dry, and shave another 5 minutes off of the drying time, so that they are 23 minutes out the door faster.
Does anyone else have any long term experience with this wash-after-wash technique. I'd like to know more about the long term effect, i.e.: whether I can indeed wash them later and be sure to remove all the traces of fix from the film emulsion.
} However, the DOS } versions of Image-Pro were discontinued quite some time ago (nearly 4 } years). Although we make our best effort to support all of our products } and customers, there have been at least 6 new Windows versions published } since the last DOS version, and the vast majority of our customers have } taken advantage of the upgrades or extended maintenance plans in order to } stay 'current'.
There is a difference in perspective about these issues for many of us.
'Nearly 4 years' is not a long time for the life of a useful product after discontinuation and, in fact, I am often loath to upgrade software if the current version is working despite the 'enhancements' of newer versions. For example, previous correspondance has revealed that IP has changed some of the ways in which it measures certain parameters and I am concerned about data compatibility. In addition, interface changes are tedious even if the newer interface is better; one has to unlearn too many things.
So, your statement that IPWin has had at least 6 versions since the last DOS version (which I have) is an indication of progress but it is also a matter of great concern about support for these lapsed versions.
} Often in our lab we are in a big rush to get prints right after the user } has finished using the EM. I've sometimes just washed the negatives } (3.25X4" Kodak EM film) just 2 minutes instead of 20, and then after } I've printed them all, I go back and wash them for 30 minutes to remove } any trace fixative that might still be remaining in the emulsion. This } way I can shave 18 minutes off of the delivery time. I also print them } with just a bit of water at their edges so that they are not completely } dry, and shave another 5 minutes off of the drying time, so that they } are 23 minutes out the door faster. } } Does anyone else have any long term experience with this wash-after-wash } technique. I'd like to know more about the long term effect, i.e.: } whether I can indeed wash them later and be sure to remove all the } traces of fix from the film emulsion. } Dear Garry, We don't use the wash-after-wash technique; rather, we wash for ~1 min in H2O, 1 min in Permawash and ~1 min in H20. This removes the fixer and negs which have been processed this way keep for several years at least. I guess if a user were *really* in a hurry, we could save ~1 min, but our way the user can take the negs home. ;-) Yours, Bill Tivol
} } } Often in our lab we are in a big rush to get prints right after the user } } has finished using the EM. I've sometimes just washed the negatives } } (3.25X4" Kodak EM film) just 2 minutes instead of 20, and then after } } I've printed them all, I go back and wash them for 30 minutes to remove } } any trace fixative that might still be remaining in the emulsion. This } } way I can shave 18 minutes off of the delivery time. I also print them } } with just a bit of water at their edges so that they are not completely } } dry, and shave another 5 minutes off of the drying time, so that they } } are 23 minutes out the door faster. } } } } Does anyone else have any long term experience with this wash-after-wash } } technique. I'd like to know more about the long term effect, i.e.: } } whether I can indeed wash them later and be sure to remove all the } } traces of fix from the film emulsion. } } } Dear Garry, } We don't use the wash-after-wash technique; rather, we wash for } ~1 min in H2O, 1 min in Permawash and ~1 min in H20. This removes the } fixer and negs which have been processed this way keep for several } years at least. I guess if a user were *really* in a hurry, we could } save ~1 min, but our way the user can take the negs home. ;-) } Yours, } Bill Tivol
Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
Please recommend, if it's possible, suitable, detailed references or your own procedure, how to isolate leukocyte-rich plasma and process it without sedimentation media, for electron microscopy.
Thanks for your understanding, Margarita Chrysanthou E-mail : cozzika-at-athena.copmulink.gr
The Department of Biological Sciences of the Exacts and Natural Sciences Faculty of the Buenos Aires University are looking for donation of a TEM and-or SEM equipment, not older than 15 years and still woorking. We will paid any handling and shipping cost. It would be very helpfull for us any other kind of assistance. Thank you vey much in advance.
Please respond to me at the addresses and numbers listed below.
Gabriel Adriano Rosa Area Microscopia Electronica, Depto. Cs. Biologicas Fac. de Ciencias Exactas y Naturales, Universidad de Buenos Aires Ciudad Universitaria, 4 piso, Pab. II, CP 1428, Buenos Aires, ARGENTINA
Cheri and all - Specific to DNA strands, I know this as the Kleinschmitt technique, now over 25 years old. The osmotically opened DNA strands are rotary shadowed. For that the specimen grids are rotated at about 60 rpm, 60-100mm from the source and at an 6-10 degree angle of incidence. The rotary shadowing makes it easier to check the continuity and to follow the under/over of the fibers. I used to evaporate Pt/Pd or for finer grain, Pt and C simultaneously. Ed Basgall's posting should help with the tungsten evaporation. Tungsten is finer still but really requires special equipment. It is possible to evaporate W - "Wolfram" in a normal evaporator, but it is slow and a lot of heat is generated. A heat shield for the specimen with a suitable aperture is advised. But why that trouble? The coarser grain from other evaporation material does not impair tracing and measuring of the DNA fibres and fibre detail is better using negative staining, or positive staining using methanolic UA. Regards Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 400+ Links, MSDS ************************ http://www.proscitech.com.au
---------- } From: Cheri Owen {cowen-at-cmb.biosci.wayne.edu} } To: Microscopy-at-sparc5.microscopy.com } Subject: Tungsten shadowing } Date: Friday, 11 July 1997 23:33 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Does anyone know a TEM protocol for "Tungsten Shadowing" say, for DNA } strand enhancement? } } Thanks! } } Cheri Owen } Detroit Neurotrauma Institute } Wayne State University } Detroit }
A Research Associate position is open in the dDpartment of Radiology, University of Texas Health Science Center at San Antonio, as the primary technician for the operation of a new atomic force microscope (Nanoscope, Digital Instruments). The person in this position would participate in studies of novel vascular biomaterial surfaces as well as in studies of vascular cell responses to mechanical forces. Applicants should have a Master's degree or a bachelor's degree with several years experience . Preferred areas of training and/or experience include scanning electron microscopy, image analysis, and cell biology. The position is currently open and we seek to hire someone as soon as possible. All applicants must apply through Human Resources, UTHSCSA. Inquiries should be directed to Dr. Gene Sprague, 210 567-5564, e-mail sprague-at-uthscsa.edu
We have used the Struers Tenupols here for many years and are generally happy with them. They wear out after a while because of all the nasty electropolishing solutions that people want to use in them but they seem to be good for at least 3-5 years (probably a lot longer if they did't get the battering that our students and RFs give them). Even then, the first things to go are the jets and these can be replaced (at a price).
You do (at least with tenupols) have to think a bit about how you will cool your electrolytes. We have a system where alcohol cooled with either dry ice or liquid nitrogen is pumped through the cooling coils on the tenupol. The temperature of the electrolyte is controlled (to give between 0 and -40 degrees C) by a homebuilt control box using the reading from a electrical resistance thermometer to control a relay that turns the pumping of the cooling alcohol on or off.
Hope this helps
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
When I had worked as a newspaper photographer back in my college days, time was always something that we were trying to cheat on. We used the EtOH wash technique to get things to dry faster.
One thing that has not been addressed is what are the photos going to be used for. Previously a typical morning at the EM would produce 80 to 100 films. We would print 8X10 images of these photos. These would later be used to keep track of the films and to keep track of the portions that would were later digitized. The printing to RC paper and processing with an Ilford developer went fairly quickly. I would hate to think of how much time it would take to send 100 several meg images to the Fugi Pixtograph printer. First there would be the storage problem, the output problem, lower resolution, and then the cost.
Dana noji-at-snowman.med.umn.edu __~0_\___ Minneapolis, Minnesota (_________) (612) 624-4687 O O http://www.tc.umn.edu/nlhome/g118/post-doc/dtn.htm
Margarita: One can simply make a buffy coat by spinning the sample in a wintrobe tube ( a skinny glass tube obtained from hematology) immediately and fixing the buffy coat by aspirating off most of the serum and replacing with glutaraldehyde/paraformaldehyde EM fixative. then cut the top and bottom off the wintrobe tube (after diamond scoring) and float this half inch long segment in the fixative until the buffy starts to get firm. Simply take a wooden applicator stick and poke the pellet out of the tube segment, process for EM and embed on edge to see all the leukocyte types in layers. bob M
We have a Fishione unit which we have used for the past fifteen years. We found the unit to be very reliable. Our unit came with a glass dish used as the reservoir. This tended to affect the response of the light detector used to stop the electropolishing when a hole appeared. In most instances however this was not a severe limitation for us. The jets and the specimen holder do need to be aligned occasionally. Also, we did have to replace the jets and the holder once since the electrodes do have a finite life. As long as the unit is rinsed thoroughly after use it should last a long time. We found the unit to be very compact and it was a simple matter to cool the electrolyte using a double beaker with the larger beaker containing the coolant. Please not that I am not familiar with the newer units and that configuration might have changed by now.
I need your suggestions on the twin-jet electropolishers for preparation of TEM samples. I need a reliable brand with reasonable price. Right now, I have information from Struers,SouthBay Technology and Fischione. Fischione has a reasonable price. Does anyone have any experience with Fischione?
Thank you for your all suggestions. Please reply my own adress.
Ibrahim Karaman Mechanical Engineering Department University of Illinois 1206 W. Green Street Urbana, IL 61801
I've never heard about Permawash. I've heard of "hypoclear" from Kodak, but the instructions state that this should be discarded after 24 hours, so it would be expensive and time consuming to make this up all the time. Is this Permawash stuff stable enough that I could keep it in a stainless steel covered tank for a week or so, to use as required?
Garry
} } Dear Garry, } } We don't use the wash-after-wash technique; rather, we wash for } } ~1 min in H2O, 1 min in Permawash and ~1 min in H20. This removes the } } fixer and negs which have been processed this way keep for several } } years at least. I guess if a user were *really* in a hurry, we could } } save ~1 min, but our way the user can take the negs home. ;-) } } Yours, } } Bill Tivol } }
It would have been my thoughts that the film need not be washed for such a long period of time, but I'm just trying to follow the instructions as supplied by Kodak as a film insert. Somehow they felt that 20 minutes was the minimum time required unless one used hypoclear or Permawash.
Garry
} Garry, } } It's probably not necessary to use the wash-after-wash technique. William } Tivol's reply probably provides the best compromise. Film, as opposed to a } print on paper, is not really very absorbent and can be washed pretty clean } pretty fast. Fix tends to rinse out readily. (I repeat, this is NOT } necessarily true for resin-coated prints, although they also wash pretty } fast, and is CERTAINLY not true for fiber-based prints.) } } As an example, in their IlfoPro newsletter, Ilford recently recommended a } wash technique for their 35mm and roll films that involved filling the } developing tank with water and inverting it 5 times, refilling it and } inverting it 10 times, and filling it a third time and inverting it 20 } times. They claim this is an archival wash technique. } } After only a couple minutes wash, I'd bet that your negatives will show no } signs of change for many years. (I can already hear the screams of } outrage!) Hypo clearing agents, PermaWash, etc., are excellent for } assuring that the negatives will outlive the people taking them, and I } certainly recommend their use for safety's sake. I still have negatives, } however, that were taken when I was 12 years old and processed in a } makeshift darkroom (it required nighttime in the country to make it } light-tight, IF the moon wasn't too bright). They were washed pretty } sloppily and still show no signs of deterioration 30 years later. } } Regarding digital imaging---in my opinion, silver-based photography is } still the most cost-effective, highest information content, easily stored } method for image CAPTURE. After capture, however, digital is more and more } the way to go for manipulation and publication. Nothing yet beats a } negative as an imaging starting point, especially considering the sizeable } investment in decent digital imaging equipment. Most labs already have } darkrooms. (I bet this will generate some comments!) } } Hope this helps. } } Randy Tindall } Center for Electron Microscopy } Southern Illinois University at Carbondale } } }
We are doing diagnostic electron microscopy as part of the Department of Pathology, in a major health care centre. The users that demand fast turn around time are the Pathologists especially when we are processing a fine needle aspirate biopsy sample. Our technique spares the patient from more invasive procedures. Because of our speed, we have been able to deliver the micrographs within 8 working hours from fresh tissue, to dispell the myth that EM as applied to biological tissue is necessarily very very slow. Often we get micrographs after 6 working hours or less. So, speed is of the essence for us. (at least that is what they tell us)
Garry
} ---------- } From: Brian G. Demczyk[SMTP:demczyk-at-erxindy.rl.plh.af.mil] } Sent: 13 July, 1997 06:35 } To: Garry Burgess } Subject: Re: RUSH PRINTS! Rush Lab. } } Just out of curiosity, what industry are you in that users } demand such a rapid turn around?! }
Sometimes when I want to make a transparency for seminars, or workshops or whatever, I put the EM negative into the enlarger and project it to a small 2X2" size on to yet another piece of EM film that I've cut in half for this purpose. (you need a special enlarger lens to get this small!). Then I cut it to size and mount it in a glass slide mount, and voila, I have supersize black and white slides, with good resolution and contrast. (at least better than 35mm projection slides) But sometimes in the past, if I tried to rush things, I notice that the image turned brown. To fix this problem though, I simply re-fix and re-wash this image, and the brown discoloration (which is probably some residual silver halide and fix) is removed, and all is well again.
Garry
} ---------- } From: rtind-at-siu.edu[SMTP:rtind-at-siu.edu] } Sent: 14 July, 1997 14:51 } To: Garry Burgess } Subject: RE: Rush Lab } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
from what i understand, Permawash is just soapy water. i simply use a mild detergent solution instead and it works fine
} I've never heard about Permawash. I've heard of "hypoclear" from Kodak, } but the instructions state that this should be discarded after 24 hours, } so it would be expensive and time consuming to make this up all the } time. Is this Permawash stuff stable enough that I could keep it in a } stainless steel covered tank for a week or so, to use as required?
} } } We don't use the wash-after-wash technique; rather, we wash for } } } ~1 min in H2O, 1 min in Permawash and ~1 min in H20. This removes the } } } fixer and negs which have been processed this way keep for several } } } years at least. I guess if a user were *really* in a hurry, we could } } } save ~1 min, but our way the user can take the negs home. ;-)
Evans blue is commonly used as a counterstain for flourescence work. It stains the specimen for observation by white light, and since it has no autoflourescence, it is invisible under the UV wavelengths.
For this application, a bottle should last an academic lifetime...I can't imagine what a case would be used for (except maybe spending surplus year-end funds)
-=W.L. Steffens=- Department of Veterinary Pathology College of Veterinary Medicine University of Georgia Athens, GA 30602 USA http://www.vet.uga.edu/vpp/wls/steffens.html
The photos are going to be used by Pathologists in a hospital for diagnostic purposes.
Garry } } One thing that has not been addressed is what are the photos going to be } used for. Previously a typical morning at the EM would produce 80 to 100 } films. We would print 8X10 images of these photos. These would later be } used to keep track of the films and to keep track of the portions that } would were later digitized. The printing to RC paper and processing with } an Ilford developer went fairly quickly. I would hate to think of how } much time it would take to send 100 several meg images to the Fugi } Pixtograph printer. First there would be the storage problem, the output } problem, lower resolution, and then the cost. } } Dana } noji-at-snowman.med.umn.edu __~0_\___ } Minneapolis, Minnesota (_________) } (612) 624-4687 O O } http://www.tc.umn.edu/nlhome/g118/post-doc/dtn.htm } } }
} I've never heard about Permawash. I've heard of "hypoclear" from Kodak, } but the instructions state that this should be discarded after 24 hours, } so it would be expensive and time consuming to make this up all the } time. Is this Permawash stuff stable enough that I could keep it in a } stainless steel covered tank for a week or so, to use as required? } Dear Garry, The bottle of Permawash says that working solution will oxidise in an open tank/tray in 6-8 hrs. A tank with a floating lid will keep for 30 days and a stoppered bottle for 90 days. The undiluted stock solution will keep a lot longer. We have our working solution in a SS covered tank like yours; I'm confident that it keeps easily for the week or so you need. Yours, Bill Tivol
Sorry to do this to you, but there was a discussion a while back about inexpensive light microscopes for children. In particular there was a recommendation from someone in the Seattle area about a $99 model (Tasco brand?). Of course, I deleted the messages but was recently asked by a friend for a recommendation. I just about went blind looking (unsuccesfully) through the June archive so it must have been further back then that! To save my eyes from further abuse, could the person posting about that microscope please just e-mail me the company name and address? I would greatly appreciate it.
Oh, also any recommendations for things to look at for kids ages 6-10 would be appreciated.
Thanks in advance.
Cheers,
John Vetrano Pacific Northwest National Laboratory P.O. Box 999 Richland, WA 99352
I need your suggestions on the twin-jet electropolishers for preparation of TEM samples. I need a reliable brand with reasonable price. Right now, I have information from Struers,SouthBay Technology and Fischione. Fischione has a reasonable price. Does anyone have any experience with Fischione?
Thank you for your all suggestions. Please reply my own adress.
Ibrahim Karaman Mechanical Engineering Department University of Illinois 1206 W. Green Street Urbana, IL 61801
We have just received an enquiry about SEM analysis for: a) pigments particles dispersed in mineral oils b) pigments particles in oil-water emulsions
Does any of you have any experience in that subject? Unfortunately, we do not count with equipment for sample preparation other than the sputter-coater and carbon evaporator.
Any help will be very welcome.
Thanks in advance,
Nora Pratta Centro Regional de Investigacion y Desarrollo Santa Fe - Argentina
} } A *very* good argument for Digital Imaging! } } Larry
Yes! but at what cost! with dwindling funds and tighter budgets all arguments are in vain......... Neelima Shah Regards... :-) :-) :-) :-) :-) :-) :-) :-) :-) :-) ;-) ;-) Visit us at http://www.med.upenn.edu/~path/core/EMCMAIN1.htm
----------------snip---------------} } 3] Is there any fixative other than Osmium which would best retain phenolics } in small root explants? } } I am not using a cryoprotectant like DMSO because I want to avoid soaking the
} roots in it, as this could allow phenolics to diffuse from the plant cells. } However, small root explants are soaked in Os-KI for several hours under } vacuum, to overnight at 4 C prior to microtomy. This allows for Os } penetration. I am relying on Os to "fix" the phenolics. Perhaps there is } something else which would penetrate more rapidly and fix or condense the } phenolics. } } Mahalo = Hawaiian for Thanks !!!!!!!! }
Dave,
have you tried RuO4? I have found it works well with a number of unsaturated ring structure compounds as a fixative. I make it up from the chloride just before use and it seems to act quite rapidly.
Advanced TEM Specimen Prep Techniques Overview: A Short Course
* Precision Angle Lapping (PAL); the =93Wedge Technique=94 * Focused Ion-Beam (FIB) Milling w. and w/o. Mechanical Pre-thinning * Materials Ultramicrotomy (MUM) * Precision Angle Cleaving (PAC)
This new one-day course provides an intensive overview of all the above techniques. The program mainly focuses on the practical aspects of the methods, the criteria for their selection, the latest developments in the= ir applications and also reviews the related instrumentation, tools, and accessories.
The next 1997 upcoming course will be offered on August 15th in Cleveland= , OH, following the MSA Annual Meeting. During the rest of this year, the course will be offered eight more times in U.S., Europe, and Asia. Pleas= e note that our regular 3-day hands-on workshops covering each one of the a= bove techniques are still offered twice a year in May and November.
To receive a copy of the short course brochure including the Registration Form and detailed information, please provide us with your complete maili= ng address, fax, and phone numbers. Thank you.
Rene E. Nicholas AMC Group (A Division of Promotech Associates, Inc.) e-Mail amcgroup2-at-aol.com
Dear Conrad } } } well I now know why I am having difficulty finding the Amoeba since } } according to the book I have just bought, at least the classic Amoeba } } Proteus is rare in the wild!!! I remember being told by a retired } } microscopist that it was rare also. } I did send a previous reply to the list. Since I got no copy I have to assume it is lost? Depending on what you see as "wild", "nature" and "natural", Amoeba is easy to find in large concentrations. At water refinery plants they are in high concentrations in the activated sludge. There is normally a buildup of material on the edge above water level, different species and even "waterbears" can be found. (Remember to use gloves collecting and handling samples.)
## [########] ## ## ## ## ## Stephan H Coeztee Electron Microscope Unit Private Bag 3 Wits 2050 South Africa
Stephan-at-Gecko.biol.WITS.ac.za
Tell: +27 11 716 2419 Fax : +27 11 339 3407
} ## [########] ## ## ## ## ## Stephan H Coeztee Electron Microscope Unit Private Bag 3 Wits 2050 South Africa
If time is of the essence have you considered doing a final wash in ethanol (70% or thereabouts)? It must be clean of course but you should knock a few more drying minutes off. Try it on something unimportant first though.
Malcolm Haswell e.m. unit University of Sunderland UK ----------
Often in our lab we are in a big rush to get prints right after the user has finished using the EM. I've sometimes just washed the negatives (3.25X4" Kodak EM film) just 2 minutes instead of 20, and then after I've printed them all, I go back and wash them for 30 minutes to remove any trace fixative that might still be remaining in the emulsion. This way I can shave 18 minutes off of the delivery time. I also print them with just a bit of water at their edges so that they are not completely dry, and shave another 5 minutes off of the drying time, so that they are 23 minutes out the door faster.
Does anyone else have any long term experience with this wash-after-wash technique. I'd like to know more about the long term effect, i.e.: whether I can indeed wash them later and be sure to remove all the traces of fix from the film emulsion.
We have been using two methods (Fe Acetate & Osmium KI) to stain for phenolics in thick (30-60u) CRYOsections of plant roots. We checked these methods on coffee leaves which have a lot of phenolics, and while both worked, the Os-KI method was superior and more specific in terms of the literature. In our research specimens (roots of alfalfa) neither of these works very well by itself. However, we get a strong staining reaction from Os-KI treated specimens if we add Toluidine Blue just prior to observation. This reveals cells with large dark deposits in their vacuoles. These could be phenolics but they could be something else. We are planning to screen several more protocols, as it would be encouraging to have several methods which yield approximately the same results, because the specificity of histochemical methods is often not understood. I have the following questions.
1] Has anyone used Toluidine Blue in consort with other treatments (especially Os-KI) to localize phenolics? In other words is this staining reaction indicative of the presence of phenolics?
2] Are there any other histochemical procedures which are known to be specific for phenolics that may be appropriate for frozen plant sections?
3] Is there any fixative other than Osmium which would best retain phenolics in small root explants?
I am not using a cryoprotectant like DMSO because I want to avoid soaking the roots in it, as this could allow phenolics to diffuse from the plant cells. However, small root explants are soaked in Os-KI for several hours under vacuum, to overnight at 4 C prior to microtomy. This allows for Os penetration. I am relying on Os to "fix" the phenolics. Perhaps there is something else which would penetrate more rapidly and fix or condense the phenolics.
We have been using two methods (Fe Acetate & Osmium KI) to stain for phenolics in thick (30-60u) CRYOsections of plant roots. We checked these methods on coffee leaves which have a lot of phenolics, and while both worked, the Os-KI method was superior and more specific in terms of the literature. In our research specimens (roots of alfalfa) neither of these works very well by itself. However, we get a strong staining reaction from Os-KI treated specimens if we add Toluidine Blue just prior to observation. This reveals cells with large dark deposits in their vacuoles. These could be phenolics but they could be something else. We are planning to screen several more protocols, as it would be encouraging to have several methods which yield approximately the same results, because the specificity of histochemical methods is often not understood. I have the following questions.
1] Has anyone used Toluidine Blue in consort with other treatments (especially Os-KI) to localize phenolics? In other words is this staining reaction indicative of the presence of phenolics?
2] Are there any other histochemical procedures which are known to be specific for phenolics that may be appropriate for frozen plant sections?
3] Is there any fixative other than Osmium which would best retain phenolics in small root explants?
I am not using a cryoprotectant like DMSO because I want to avoid soaking the roots in it, as this could allow phenolics to diffuse from the plant cells. However, small root explants are soaked in Os-KI for several hours under vacuum, to overnight at 4 C prior to microtomy. This allows for Os penetration. I am relying on Os to "fix" the phenolics. Perhaps there is something else which would penetrate more rapidly and fix or condense the phenolics.
It's probably not necessary to use the wash-after-wash technique. William Tivol's reply probably provides the best compromise. Film, as opposed to a print on paper, is not really very absorbent and can be washed pretty clean pretty fast. Fix tends to rinse out readily. (I repeat, this is NOT necessarily true for resin-coated prints, although they also wash pretty fast, and is CERTAINLY not true for fiber-based prints.)
As an example, in their IlfoPro newsletter, Ilford recently recommended a wash technique for their 35mm and roll films that involved filling the developing tank with water and inverting it 5 times, refilling it and inverting it 10 times, and filling it a third time and inverting it 20 times. They claim this is an archival wash technique.
After only a couple minutes wash, I'd bet that your negatives will show no signs of change for many years. (I can already hear the screams of outrage!) Hypo clearing agents, PermaWash, etc., are excellent for assuring that the negatives will outlive the people taking them, and I certainly recommend their use for safety's sake. I still have negatives, however, that were taken when I was 12 years old and processed in a makeshift darkroom (it required nighttime in the country to make it light-tight, IF the moon wasn't too bright). They were washed pretty sloppily and still show no signs of deterioration 30 years later.
Regarding digital imaging---in my opinion, silver-based photography is still the most cost-effective, highest information content, easily stored method for image CAPTURE. After capture, however, digital is more and more the way to go for manipulation and publication. Nothing yet beats a negative as an imaging starting point, especially considering the sizeable investment in decent digital imaging equipment. Most labs already have darkrooms. (I bet this will generate some comments!)
Hope this helps.
Randy Tindall Center for Electron Microscopy Southern Illinois University at Carbondale
Herbert McLean Evans was chair of Anatomy at Univ of Calif Berkeley, from 1915. His early research included use of vital dyes for studies on blood volume, macrophages, ovarian estrous cycle, etc. In a 1920 paper (Dawson, Evans, Whipple. "Blood volume studies. III. Behavior of large series of dyes introduced into the circulating blood." Amer J Physiol 51:232-256) it was shown that a blue azo dye (T-1824) was "slightly superior" for blood volume measurement than the vital red series previously used. The dye came to be know as "Evans blue." Criteria that were important were: (1) non-toxic, (2) not stored in tissue, (3) color allows accurate determination, (4) removed quite slowly from the blood stream.
An example of more recent use: Bergh, Damber, 1988, Int J Androl 11:449.
Kent (A. Kent Christensen, Dept of Anatomy and Cell Biology, University of Michigan Medical School), {akc-at-umich.edu} )
------------------------------
On Sun, 13 Jul 1997, Gilbert T. Groehn wrote:
} Our lab acquired a case of EVANS BLUE stain } from one of our clients and we can not locate } this in any of our chemistry manuals. } } What are the normal uses for this stain (eg. } histology, hematology, etc.) ? We would be } using it only for light microscopy. } } What is the standard mixing ratios and } formulas for various applications? } } Any help on this item would be most appreciated. } } TIA } Gil Groehn } ULTRAMED, INC. } } ============================================================= } ULTRAMED, INC. Research Div. ad408-at-detroit.freenet.org } Grosse Pointe Farms, MICH 48236 USA Biomedical Consultants } =============================================================== }
We use the spot mode pretty routinely to identify inorganic particles in our polyester films. The size of these particles is anywhere from about 0.5 =B5m to 10 =B5m. So far it has worked quite well for us. When we = switch back to full scan after using the spot mode, we invariably see a "burn" mark which is exactly where we think it ought to be. We often look at a 2nd spot away from the structure that interests us to be sure that we're not just picking up a background.
} I travel the world teaching practical electron microscopy and it = worries me } the emphasis that people place on "Spot Mode". I do not teach the use = of } spot mode for the following reasons-
} 1. Just because the spot appears in a certain position on the = screen } is this its correct position. I have found machines many microns out = of } step in X and Y directions.
} 2. Specimens charge and switching out of spot mode does not give = the } operator an easy opportunity to see if the spot had moved during the } analysis.
} 3. Spot mode gives a false sense of analytical volume, no matter = how } small that spot may be we are almost certainly evaluating microns of } material.
} Do others worry about spot mode accuracy, do others test the spot mode } accuracy? Is it not better to simply increase the magnification = watching } the area of interest all the way up before the analysis and all the way } down after the analysis?
Youre vibration problem resembles a problem I had about 7 years ago when I was working with scanning tunneling microscopy at the university of Nijmegen (The Netherlands). Our STM in a UHV system was on the 4th floor, so we had some trouble with vibrations, mainly stemming from air ventilation compressors in the base of the building. In the same basement there were three vibration free sites consisting of concrete bloks (2x2x2 m) placed in sand an having no connection to the foundation of the building. We measured the vibrations on these bloks and found that they were only a little bit better than on the 4th floor!! We found out that the vibrations from the compressors propagates not only through the building construction but also through the ground or sand. So the effect of de-coupling the concrete blocks from the building foundation did not eliminate the vibration-coupling, but it did reduce the (effective) mass of the blocks. Hence the vibration amplitude growths! The foundation of the building, although directly coupled to the floors of the compressors, showed a quit clean vibration spectrum, because the mass is high (effectively the whole building).
So, back to your question: cutting the floor around your microscope will only help if this floor is the only vibration coupling to the microscope. If also the ground (dirt) or air (acoustic) couples vibrations to your microscope, the end result will be worse, since the mass of your system is reduced.
Success, Bart
Bart Nelissen
Akzo Nobel Central Research
Dept of Applied Physics, Microscopy
Tel: +31 26 366 1371
Fax: +31 26 366 5272
eMail: Bart.B.J.Nelissen-at-Akzo.nl
Note: Do not use automatic reply or answer facilities in your e-mail program
to respond to this message (the sender-address is distorted by our 'gate-keeper').
I remember many years ago trying to clean some grid boxes (the white LKB type with a clear lid) with ethanol and it all went disastrously wrong. I can't remember whether both types of plastic deteriorated but the boxes were unusable. It may be that SPI boxes are made of similar material - try asking them. I now just give boxes a wash with soapy water (maybe a quick ultrasonic clean) and rinse in distilled water then dry for a couple of days I don't know if this helps.
Malcolm Haswell e.m. unit University of Sunderland UK ----------
O.K. you professionals here's a real puzzler for you:
I have a user here who has about 24 SPI Slide-A Grid Boxes. They were a little dusty (they may have been used before - heavens not to say they didn't come absolutely pristine from SPI), and so he throughly washed them and rinsed them with Ethanol, only now he can not get them back open any longer! They are really stuck.
I don't know if he removed some type of lubication on the box, maybe the "natural" lube from the manufacturing process, or if he caused the plastic to swell with the EtOH, but I could certianly use any suggestion. Maybe we should throw this up to the group - surely someone has cleaned grib boxes before.
We can get the box lids to move upto 5mm or so but then they stop and with work we can get them to move back, but we still have freed any lids. We've tried prying up the lids a little from the back edges (where they slide along rasied ridges) - didn't help much. We've even tried soaking them again in water or EtOH hoping to provide some "lubrication" (there are no grids in the boxes, but didn't really want to soak them in some thing oily) - still with no success.
I oringinally asked this of Charles Garber (of SPI) and neither of us have come up with any ideas, but he did throw in the following:
Of the box parts:
} The plastic is an antistatic formulation however, alcohol being so } polar is not going to swell the base piece at least. If the alcohol } was hot or if it was in contact with the plastic for some extended } period of time, then who knows, but I just can not believe that the } alcohol would have done anything to it in the way you are } suggesting. } } The sliding top might be another story. I forgot the plastic that is } used for it, but it might be a clear PVC. But even then, room } temperature alcohol should not have had time to do anything. }
Any suggestions?
The things fools will do ....
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513-529-5712 Fax: 513-529-4243 E-mail: edelmare-at-muohio.edu
"640K ought to be enough for anybody." -- Bill Gates, 1981
} } Can any of our German colleagues help with translation please? Came } across the term PHOSPHORWOLFRAM acid in a technical paper. Suspect it is } phosphoric acid but want to be sure before proceeding. } Thanks in advance } Ronnie Houston I think you will find that it is phosphotungstic acid (hence the atomic symbol W for tungsten)
Chris
Chris Gilpin Biological Sciences Electron Microscope Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 161 275 5170 fax +44 161 275 5171 http://www.biomed.man.ac.uk/biology/emunit/emhome.html
I have a client who will be purifying a retrovirus using a sucrose gradient.
The question is what medium should the sample be in to do the negative stain. Sucrose? Buffer? If anyone has worked with retrovirus I would like to hear your suggestion.
Thank you in advance
Karen Vaughn,EM Technician phone: (352)392-1184 University of Florida fax: (352)846-0251 Electron Microscopy Core Lab. e-mail: klv-at-biotech.ufl.edu 214 Bartram Hall web: http://www.biotech.ufl.edu/~emcl/ Gainesville, Fl. 32611
I'm not sure what is causing the brown staining, especially since you indicate that it's temporary. My GUESS is that it's inadequate fixing time or exhausted fixer. What are your fixing times?
Randy,
Usually I fix for 2-3 minutes, but in this case, it might be more, since the slides were actually just piled up in a staining beaker filled with hypo. Since the slides were in contact with each other, I'm quite sure that this explains the inadequate fixation. Actually usually I look at them under the safelight to ensure that they are adequately fixed, but sometimes, when I had a huge volume, then I must have missed a couple, because I had so many. (about 200 or so)
I use some sort of Kodak fixer, but I can't remember the name right at this second. I usually do not use hypochek, because as I've said, usually it's obvious when the fixer is losing it's strength, because it starts to take significantly longer than 2 minutes to fix our EM negatives. Because we work under safelight, this is something that can be checked each and every time that we develop film.
What fix do you use? Do you check your fix for exhaustion with HypoChek or a similar product that gives you a white precipitate when the fixer is saturated with silver compounds? Interesting.
I agree with you about the confusion about Permawash. It would have to be something more chemically like Hypo Clear than soap, because I can't imagine a soapy solution removing fix from the emulsion, though it's an interesting thought. However, I also don't think that I'd like a soapy solution as a substitute for Photoflo, because I think that a soapy solution might leave a residue, which is probably the last think that one would want in the last step of their film developing.
Incidentally, someone on the server said that they thought PermaWash was a soap solution. I think they're confusing it with PhotoFlo, which, as you know, is just a wetting agent to prevent water drops from marking the film when it dries. PermaWash is more like Hypo Clear in that it chemically removes fixer or changes it into more soluble, harmless compounds (I forget which!). Another very good product is Orbit Bath, which is very cheap and effective, but unfortunately seems to be now sold under a different name. Ask in a good photo shop, if you're interested. There is also Hypo Eliminator, another Kodak product, which the Kodak folks once told me is different than Hypo Clear and, I believe, is meant strictly for films, rather than both film and paper.
The history of these compounds seems to be kind of interesting, because I've read somewhere that ordinary sea water works as a hypo clearing agent and that it forms the basis for the modern products. Take it for what it's worth...
Neat method of making slides, by the way. I'm going to remember it for possible future use.
Yes, it works fine. But the biggest obstacle for someone doing this for the first time is to make sure that they have a lens that is capable of making such a small focused image. You also have to make sure that you use glass slide holders, because negative film cannot stand up to the heat of a slide projector without glass protection, because it's not as tough as slide film.
Garry
} ---------- } From: rtind-at-siu.edu[SMTP:rtind-at-siu.edu] } Sent: 15 July, 1997 09:08 } To: Garry Burgess } Subject: RE: Rush Lab + Projection Slides } } Garry, } } I'm not sure what is causing the brown staining, especially since you } indicate that it's temporary. My GUESS is that it's inadequate fixing time } or exhausted fixer. What are your fixing times? } } Randy, } } Usually I fix for 2-3 minutes, but in this case, it might be more, since the } slides were actually just piled up in a staining beaker filled with hypo. } Since the slides were in contact with each other, I'm quite sure that this } explains the inadequate fixation. Actually usually I look at them under the } safelight to ensure that they are adequately fixed, but sometimes, when I had } a huge volume, then I must have missed a couple, because I had so many. } (about 200 or so) } } I use some sort of Kodak fixer, but I can't remember the name right at this } second. I usually do not use hypochek, because as I've said, usually it's } obvious when the fixer is losing it's strength, because it starts to take } significantly longer than 2 minutes to fix our EM negatives. Because we work } under safelight, this is something that can be checked each and every time } that we develop film. } } } What fix do you use? Do } you check your fix for exhaustion with HypoChek or a similar product that } gives you a white precipitate when the fixer is saturated with silver } compounds? Interesting. } } I agree with you about the confusion about Permawash. It would have to be } something more chemically like Hypo Clear than soap, because I can't imagine } a soapy solution removing fix from the emulsion, though it's an interesting } thought. However, I also don't think that I'd like a soapy solution as a } substitute for Photoflo, because I think that a soapy solution might leave a } residue, which is probably the last think that one would want in the last } step of their film developing. } } Incidentally, someone on the server said that they thought PermaWash was a } soap solution. I think they're confusing it with PhotoFlo, which, as you } know, is just a wetting agent to prevent water drops from marking the film } when it dries. PermaWash is more like Hypo Clear in that it chemically } removes fixer or changes it into more soluble, harmless compounds (I forget } which!). Another very good product is Orbit Bath, which is very cheap and } effective, but unfortunately seems to be now sold under a different name. } Ask in a good photo shop, if you're interested. There is also Hypo } Eliminator, another Kodak product, which the Kodak folks once told me is } different than Hypo Clear and, I believe, is meant strictly for films, } rather than both film and paper. } } The history of these compounds seems to be kind of interesting, because } I've read somewhere that ordinary sea water works as a hypo clearing agent } and that it forms the basis for the modern products. Take it for what it's } worth... } } Neat method of making slides, by the way. I'm going to remember it for } possible future use. } } Yes, it works fine. But the biggest obstacle for someone doing this for the } first time is to make sure that they have a lens that is capable of making } such a small focused image. You also have to make sure that you use glass } slide holders, because negative film cannot stand up to the heat of a slide } projector without glass protection, because it's not as tough as slide film. } } Garry } }
} Sorry to do this to you, but there was a discussion a while back about } inexpensive light microscopes for children. In particular there was a } recommendation from someone in the Seattle area about a $99 model (Tasco } brand?). Of course, I deleted the messages but was recently asked by a friend } for a recommendation.
Younger children do MUCH better with an erect image, so the first decision is "dissecting" vs. compound. And another choice is monocular vs. binocular. Young children have problems with both the eye spacing and the convergance required by binocs, so the extra cost of two oculars may not be justified. } } Oh, also any recommendations for things to look at for kids ages 6-10 would be } appreciated.
Children want to DO things, rather than just look. My grandson was fascinated with watching epsom salt crystals grow when he was 6. You'll find a wealth of suggestions in the Project MICRO bibliography (see below); look in section IIA, "The microscopic world".
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
} Date: Mon, 14 Jul 1997 15:54:34 -0500 } From: David Knecht {knecht-at-uconnvm.uconn.edu} } To: microscopy-at-sparc5.microscopy.com } Subject: ConA-labeled } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Does anyone know any good sources besides Sigma for conA labeled with } Rhodamine, Texas Red or another non-FITC dye. Thanks- Dave } } Dr. David Knecht } Department of Molecular and Cell Biology } University of Connecticut } U-125 } Storrs, CT 06269 } Knecht-at-uconnvm.uconn.edu } Linscott's Directory of Immunological and Biological Reagents (ISBN: 0-9604920-4-6, Phone: 415-544 9555, about $70) is a fantastic reference that lists almost every antibody made by every company. It is a valuable book I recommend for anyone doing immunostaining.}
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Hello Gilbert and cyber-folk Evans blue can also be used to determine cellular integrity in plant cell suspensions, squashes, etc. The dye will not penetrate intact plasmalemma, while staining damaged cells.
Yet another use for your case of stain! Have you considered a "garage sale"?
James Wesley-Smith EM Unit University of Natal Durban, South Africa
------------------------------
On Sun, 13 Jul 1997, Gilbert T. Groehn wrote:
} Our lab acquired a case of EVANS BLUE stain } from one of our clients and we can not locate } this in any of our chemistry manuals. } } What are the normal uses for this stain (eg. } histology, hematology, etc.) ? We would be } using it only for light microscopy. } } What is the standard mixing ratios and } formulas for various applications? } } Any help on this item would be most appreciated. } } TIA } Gil Groehn } ULTRAMED, INC. } } ============================================================= } ULTRAMED, INC. Research Div. ad408-at-detroit.freenet.org } Grosse Pointe Farms, MICH 48236 USA Biomedical Consultants } =============================================================== }
--IMA.Boundary.790299868 Content-Type: text/plain; charset=US-ASCII Content-Transfer-Encoding: 7bit Content-Description: cc:Mail note part
Mike,
It would be interesting to know a situation where this enlarger would be the preferred method of producing prints as opposed to taking the negatives that need dodge/burn, scan them and then digitally "correct" them? Good luck on finding this enlarger used etc,!
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello Everyone:
Does anyone know where I can get a used LogEtronics 55-EM enlarger (or something similar with auto-dodge & burn-in capabilities)in good working condition?
Thanks,
Michael Coviello Lab Manager The University of Texas -at- Arlington --IMA.Boundary.790299868 Content-Type: text/plain; charset=US-ASCII; name="RFC822 message headers" Content-Transfer-Encoding: 7bit Content-Description: cc:Mail note part Content-Disposition: inline; filename="RFC822 message headers"
Received: from ns2.baxter.com (159.198.1.38) by ccmailgw.mcgawpark.baxter.com
(IMA Internet Exchange 2.1 Enterprise) id 00142195; Fri, 11 Jul 97 12:42:20 -0500 Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by ns2.baxter.com (8.8.0/8.8.0) with SMTP id MAA15639; Fri, 11 Jul 1997 12:48:58 -0500 (CDT) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id JAA19418 for dist-Microscopy; Fri, 11 Jul 1997 09:53:28 -0500 Received: from mae.uta.edu (mae.uta.edu [129.107.25.180]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id JAA19415 for {Microscopy-at-sparc5.microscopy.com} ; Fri, 11 Jul 1997 09:53:27 -0500 Received: from ns1 ([129.107.25.216]) by mae.uta.edu (8.8.4/8.8.4) with SMTP id JAA02515 for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 11 Jul 1997 09:57:39 -0500 Message-Id: {3.0.32.19970711100029.0091e930-at-mae.uta.edu} X-Sender: coviello-at-mae.uta.edu X-Mailer: Windows Eudora Pro Version 3.0 (32)
} from what i understand, Permawash is just soapy water. i simply use a } mild detergent solution instead and it works fine } You may be thinking of Photoflo - which is essentially a wetting agent, a detergent - used to break the hydrophobicity of film and permit sheeting of the water. This is similar to the wetting agents used in dishwashing machines (e.g., for spotless glassess). Permawash should contain some chemical scavengers used to remove resisual fixers in the film/papers. Any photo-chemical types listening to this?
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
I am interested in the size of particle that you talk about, what densit= y do you see in your material having carried out the analysis?
For materials similar to carbon the volume of material involved with an electron beam of an energy suitable for most EDX analysis (say 15kV) wil= l be in excess of 2.7 microns? To carry out an analysis of 0.5 micron structures with material the density of carbon would require a beam energ= y of less than 6kV or for iron less than 10kV.
The mark you call a "burn" is this not really contamination? There is usually no damage to a specimen if you see a dark mark, contamination. T= he problem is that with your materials you may well drive off the volatile components and then you will burn out an area of specimen, the depth dependant upon the time of your observation or analysis.
However you seem to have conclusive proof that you are analysing the area=
that you have made your target. My point was not that a spot analysis is=
taboo but that we should not blindly use a spot analysis without knowing more about its accuracy. Some replies have taken the attitude that becau= se they have used the technique for many many years there cannot be a proble= m! Unless you know exactly what is happening in a microscope I do not agree=
that this attitude is sufficient, I can show you instruments where this attitude would be a disaster.
In the consultancy business the biggest problem that one confronts is the=
"scientist" who has been doing things his way for 20 years and will not change. I understand that we may well compare driving a microscope to driving a car. Who would by a new car and then go on a course to learn t= o drive it? The same approach in SEM however would be constructive; new instruments open up new techniques and new areas of investigation. From what I see the topic of scanning electron microscopy changes considerably=
within a two year cycle and operators who persist in using their 20 year old techniques, even on an old instrument, are somewhat lost! The "I onl= y use 25kV" and " an SEM will not work above 5,000X" bunch really worry me and they should also worry their supervisors! =
Is a true scientist someone who experiments, most SEM operators do not! =
This is not a one country thing it is the same world wide, just because w= e are microscopists we seem to forget we are also scientists. What do the masses think?
} Now, with negative, flatbed scanners one could scan the negative (10 } minutes) and print an image on transparency paper with an inkjet } (Epson 1440 dpi or a dyesub printer). Haven't tried this with TEM } negatives but I know people who do. } I have no doubt that this would work, but it requires yet more capital equipment. And here in Manitoba Canada, the money is NOT AVAILABLE. Hence, I have to dismiss all the talk about digital images and scanners, and that sort of thing. It's something like arguing that a Mercedes is a nice car. I have no doubts that it is, but I nevertheless won't be driving one in the near future, unless I win a rather large lottery. } Seems like about one hour would do it. } } What about a positive TEM film that you could project directly. } I have not heard about the existence of this sort of TEM film, but even if we did have the film, we would still suffer from the fact that we would have to crop a considerable amount out of the frame in order to get it to fit our slide projector, even with the supersize size. (ie: 2X2")
Secondly, we usually don't know ahead of time that we will need the transparencies. The urgent images are the 8X10" prints, of which I'm rushed to produce as fast as possible. There is no time for a Pathologist to scope the case twice, once with positive film. And in any event, we would need a record on negative film in any event.
I'm sure that I could develop this normal TEM film in a reversal sort of way, in the same manner that one might work with T-MAX film developed with reversal chemistry, but this produces positives directly, and there is no negative produced, so that I would have the above problems, plus the cropped images.
It seems to be quite satisfactory to produce my positive slides the way I do, and usually speed is not a big factor with these positive images.
Garry } } } } } ______________________________ Reply Separator } _________________________________ } Subject: RE: Rush Lab + Projection Slides } Author: GBurgess (GBurgess-at-exchange.hsc.mb.ca) at unix,mime } Date: 7/14/97 2:02 PM } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} the biggest problem that one confronts is the "scientist" who has been = doing } things his way for 20 years and will not change.
That's certainly not the case for me! I'm a chemist who was been partially using myself and partially supervising a new SEM for less than a year. I've subscribed to the list in the hopes that I'd learn something.
} I am interested in the size of particle that you talk about, what = density } do you see in your material having carried out the analysis?
} For materials similar to carbon the volume of material involved with = an } electron beam of an energy suitable for most EDX analysis (say 15kV) = will } be in excess of 2.7 microns? To carry out an analysis of 0.5 micron } structures with material the density of carbon would require a beam = energy } of less than 6kV or for iron less than 10kV.
I guess I didn't express myself very clearly. We're looking at low-level amounts of inorganic particulate contaminants or additives in plastic (C,H,O) and what we want is QUALITATIVE information, such as "Does this particle contain calcium or does it contain silicon?" We wouldn't dare to try say anything about amounts (other than perhaps "lots" or "hardly any") because we're never sure how much of the polymer matrix we're hitting along with the particle. (The particles are at varying depths.)
In my--admittedly still amaturish--opinion, spot analysis can be OK if what you want is qualitative information. You have to make sure that the customers don't think the results are in any way quantitative. (They always want to and you have to keep on banging on 'em.)
} The mark you call a "burn" is this not really contamination? There is } usually no damage to a specimen if you see a dark mark, contamination.
I take the placement of the "burn" mark as proof that the positioning of the beam in spot mode is OK. As to what the dark "burn" mark actually is, I'm open. (I put those quotes on the word for a reason!) However I think it's thermal degradation of the polymer, which melts around 260=B0 = C and softens and shrinks before that. I think this because we don't get these marks when we look at metals, etc.
Cindy Bennett Hoechst Diafoil
Disclaimer: these are my opinions only and not those of my company.
we are drifting well away from the main theme, but you can still make slides with your enlarger even if you haven't got close focussing on your lens. It's more fiddly but if you have a light box you simply put the e.m. negative on the light box under the enlarger and the film to be exposed in the negative carrier. The tricky bit is getting the position and focus right but you do most of that by enlarging something of the right format first onto the light box. Then of course when you're exposing the film in the enlarger you must remember to turn on the light box and not the enlarger.
I have used this a couple of times and it works fine in an emergency although of course if you were doing a lot it would be easier to make the slides with a close-up 35mm camera and a light box..
Malcolm Haswell e.m. unit University of Sunderland UK ----------
{SNIP} Neat method of making slides, by the way. I'm going to remember it for possible future use.
Yes, it works fine. But the biggest obstacle for someone doing this for the first time is to make sure that they have a lens that is capable of making such a small focused image. You also have to make sure that you use glass slide holders, because negative film cannot stand up to the heat of a slide projector without glass protection, because it's not as tough as slide film.
I will tried the glow discharge and the carbon film to change the hydrophobicity...and also putting my grids (specimen and stain) in the oven 60C for 24 hours before looking at them to get rid of possible water trapping. I really appreciate your help, especially during summer time when a lot of people are absent...
thanks again,
Diane Montpetit microscopist Food research center agro alimentaire canada st-hyacinthe, quebec, canada tel 514 773 1105 fax 514 773 8461 e mail montpetitd-at-em.agr.ca
First, someone working 20 years in the industry is, by definition, successful. This means that he/she is providing information that is useful to others in solving their problems.
Second, our employers could care less about kv's, resolution, magnification, etc. Their interest is in solving problems. Doing the same thing for 20 years (even with a 20 year old microscope) may be what it takes. I have seen old-timers with a 20 year old scope work circles around newbies with newbie microscopes.
Finally, I agree whole-heartedly with Steve--we must be open to new ideas to solve difficult problems. I worked for a major computer manufacturer to solve a circuit board plating problem (wavy circuit traces). They ALWAYS USED 25 kv to examine the photo resist on the boards that produced the circuit traces. At 25 kv they looked straight and clean. I showed them what they looked like at 5kv and at 2kv. The edges were full of unremoved resist film whose pattern matched the wavy lines identically.
Providing INFORMATION to solve problems is our goal.
Jim Harper Amoco Fabrics and Fibers ______________________________ Reply Separator _________________________________
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am interested in the size of particle that you talk about, what density do you see in your material having carried out the analysis?
For materials similar to carbon the volume of material involved with an electron beam of an energy suitable for most EDX analysis (say 15kV) will be in excess of 2.7 microns? To carry out an analysis of 0.5 micron structures with material the density of carbon would require a beam energy of less than 6kV or for iron less than 10kV.
The mark you call a "burn" is this not really contamination? There is usually no damage to a specimen if you see a dark mark, contamination. The problem is that with your materials you may well drive off the volatile components and then you will burn out an area of specimen, the depth dependant upon the time of your observation or analysis.
However you seem to have conclusive proof that you are analysing the area that you have made your target. My point was not that a spot analysis is taboo but that we should not blindly use a spot analysis without knowing more about its accuracy. Some replies have taken the attitude that because they have used the technique for many many years there cannot be a problem! Unless you know exactly what is happening in a microscope I do not agree that this attitude is sufficient, I can show you instruments where this attitude would be a disaster.
In the consultancy business the biggest problem that one confronts is the "scientist" who has been doing things his way for 20 years and will not change. I understand that we may well compare driving a microscope to driving a car. Who would by a new car and then go on a course to learn to drive it? The same approach in SEM however would be constructive; new instruments open up new techniques and new areas of investigation. From what I see the topic of scanning electron microscopy changes considerably within a two year cycle and operators who persist in using their 20 year old techniques, even on an old instrument, are somewhat lost! The "I only use 25kV" and " an SEM will not work above 5,000X" bunch really worry me and they should also worry their supervisors!
Is a true scientist someone who experiments, most SEM operators do not! This is not a one country thing it is the same world wide, just because we are microscopists we seem to forget we are also scientists. What do the masses think?
Hello all. last week I stupidly stuck a Cu grid on top of the area of interest of a TEM cross section. Having tried to remove it for a few days, I thought I'd turn to the microscopy community for some help! The sample is Si, polished to less than ten microns thick (orangey colour), with Al and SiO2 on the top surface. It's the only one I have. I stuck the 2x1mm slot grid onto the sample with 5-minute epoxy (devcon); I didn't realise the grid was in the wrong place until a couple of hours later. Since then I've soaked it in dmf (dimethylformamide) [3 days] acetone [a few hours] and ashed it in nitrogen and oxygen [a couple of hours]. The sample is still in one piece and stuck to the grid. I've tried pushing it about with a fine hair but it's still well fixed. So, while it is soaking for a little longer in dmf and being ashed periodically, has anyone got any bright ideas how to rescue my sample?
Many thanks in advance,
Richard Beanland, Gmmt Ltd., Caswell, Towcester, Northants NN12 8EQ
Tel +44 1327 356363 Fax +44 1327 356775 e-mail richard.beanland-at-gecm.com
For all who are interested in burns in polymers under SEM, one does not only get them in Spot Mode, but when looking at high magnification in raster mode. Strange things can happen when looking at banded spherulites of polyethylene, for example: because of the periodic variations in crystal orientation, one gets mass transport (not mass transit, which is the underground railway in Hong Kong!) and the banded spherulitic structure appears to "develop" as if by etching, but what appears is in fact an artifact which mimics the real thing.
Similar developments occur when trying to look at these beasties directly in sections under the TEM, which is one reason why staining (chlorosulphonic acid, RuO4) and etching (permanganic) techniques were developed.
There is a rather poor quality picture of a banded spherulite on my home page: URL as in the signature, but to go straight there type:
I have looked through all of my EM catalogues, checking the section on Photography for anything like Permawash, but the only product that I can find that comes close to that idea of hypo remover is the Kodak product Hypo Eliminator. My problem with the Hypo Eliminator is that the instructions suggested that the working solution only lasts 24 hours, but I wanted to keep it in a stainless steel tank for about a week, and just use it conveniently, without having to make up a working solution every day.
That said, providing there were no water marks on the film, I'm still wondering if I can rewash it later for 20 minutes, after the negatives have already dried, and effectively remove any fixer that might have been left in the emulsion the first time for a very abbreviated wash.
Other people have suggested that Kodak was being very conservative with their wash times, but I'm not sure if this is true. Kodak had no difficulty shortening the wash times for RC paper when they felt that a 2 minute wash time was sufficient for this photographic paper. So, if they really felt that 2 or 3 minutes wash time was sufficient for their EM film, then I would suspect that that would be their recommendation.
Garry } } } from what i understand, Permawash is just soapy water. i simply use a } } mild detergent solution instead and it works fine } } } You may be thinking of Photoflo - which is essentially a wetting agent, a } detergent - used to break the hydrophobicity of film and permit sheeting } of the water. This is similar to the wetting agents used in dishwashing } machines (e.g., for spotless glassess). Permawash should contain some } chemical scavengers used to remove resisual fixers in the film/papers. Any } photo-chemical types listening to this? } } } #################################################################### } John J. Bozzola, Ph.D., Director } Center for Electron Microscopy } Neckers Building, Room 146 - B Wing } Southern Illinois University } Carbondale, IL 62901-4402 } 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 thread of this conversation appears to be drifting toward making projection slides from EM negatives.
The technique that I have used for years is to place the EM negative (SEM or TEM) on a light table with a cardboard mask around the negative to block peripheral light. Use Kodak Technical Pan film in a standard 35 mm camera with a macro lens. Bracket exposures 1/2 f-stop and develop for maximum contrast. (From start to mounted slides, less than 1 hour).
One gets crisp slides with perfect contrast (assuming that the original negatives were good). The advantage is that one can crop the EM negative by adjusting distance of the camera from the negative. Disadvantage is that one cannot label the slide unless one is willing to put rub-on letters on the EM negatives (I don't).
______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 The College of New Jersey fax: (609) 771-2674 Trenton, NJ 08650-4700
Taking epoxy off without damaging the silicon or aluminum is tough. Acetic acid will work but may damage the silicon film. It does not attack bulk aluminum (much) but at the thin film level it might. Methyl cloride is also useful for dissolving epoxy--some paint removers will also dissolve epoxy.
best regards mark
Mark W. Lund, PhD Director } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"The state is good at simple tasks, like killing people and seizing their wealth. It has far more trouble reaching inside individuals and making them good." Doug Bandow
Malcolm, This sounds bizarre. Are you talking about a contact print here on to film? Or do you mean leave the negative carrier in the carrier holder of the enlarger, and use it upside down? (sort of like using the enlarger as an upside down camera.)
Garry
} Garry } } we are drifting well away from the main theme, but you can still make slides } with your enlarger even if you haven't got close focussing on your lens. } It's more fiddly but if you have a light box you simply put the e.m. } negative on the light box under the enlarger and the film to be exposed in } the negative carrier. } The tricky bit is getting the position and focus right but you do most of } that by enlarging something of the right format first onto the light box. } Then of course when you're exposing the film in the enlarger you must } remember to turn on the light box and not the enlarger. } } I have used this a couple of times and it works fine in an emergency } although of course if you were doing a lot it would be easier to make the } slides with a close-up 35mm camera and a light box.. } } Malcolm Haswell } e.m. unit } University of Sunderland } UK } ---------- } From: Garry Burgess } To: 'Microscopy Society of America } Subject: RE: Rush Lab + Projection Slides } Date: 15 July 1997 18:12 } } {SNIP} } Neat method of making slides, by the way. I'm going to remember it for } possible future use. } } Yes, it works fine. But the biggest obstacle for someone doing this for the } first time is to make sure that they have a lens that is capable of making } such a small focused image. You also have to make sure that you use glass } slide holders, because negative film cannot stand up to the heat of a slide } projector without glass protection, because it's not as tough as slide film. } } Garry } }
Hi Richard, Acetone disolves araldite so it seems a good start. Try soaking some of your cured epoxy (not your specimen) in acetone for a while and see if it softens, if it does then soak your specimen in it for longer. It is probably very difficult for the acetone to get right under the copper grid so it may need quite a long time. Plasma ashing seems to spread epoxy around the specimen but does not appear to remove it.
Good luck.
Ron =========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
To get slides with labelling, I overlay a piece of acetate with the letters, arrows etc. rubbed onto it, placed carefully on top of the negative. Provided the acetate is clean, it works very well. Of course, my negatives are 3in x 4in; it might be more difficult with 35mm film.
Lesley Weston.
On Wed, 16 Jul 1997, Donald Lovett wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } The thread of this conversation appears to be drifting toward making } projection slides from EM negatives. } } The technique that I have used for years is to place the EM negative (SEM } or TEM) on a light table with a cardboard mask around the negative to } block peripheral light. Use Kodak Technical Pan film in a standard } 35 mm camera with a macro lens. Bracket exposures 1/2 f-stop and develop } for maximum contrast. (From start to mounted slides, less than 1 hour). } } One gets crisp slides with perfect contrast (assuming that the original } negatives were good). The advantage is that one can crop the EM negative } by adjusting distance of the camera from the negative. Disadvantage is } that one cannot label the slide unless one is willing to put rub-on } letters on the EM negatives (I don't). } } ______________________________________________________________________ } Donald L. Lovett e-mail: lovett-at-tcnj.edu } Assoc. Professor, Dept. of Biology voice: (609) 771-2876 } The College of New Jersey fax: (609) 771-2674 } Trenton, NJ 08650-4700 } } } }
1. How long can I store rat tissue in glutaraldehyde fixative before conpleting the tissue preparation process and not experience tissue degradation? Can I go as long as three weeks? At this point, I don't know if I will be processing for SEM (CPD) or TEM - depends on the LM results.
2. Should I store in buffer rather than the fixative? Some other solution?
3. Would I use a different formulation of fixative for this kind of delayed processing of tissue?
So much for quick questions and thanks so much for your help! Now, I have to go answer someone else's question.
1. How long can I store rat tissue in glutaraldehyde fixative before conpleting the tissue preparation process and not experience tissue degradation? Can I go as long as three weeks? At this point, I don't know if I will be processing for SEM (CPD) or TEM - depends on the LM results.
2. Should I store in buffer rather than the fixative? Some other solution?
3. Would I use a different formulation of fixative for this kind of delayed processing of tissue?
So much for quick questions and thanks so much for your help! Now, I have to go answer someone else's question.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Richard:
I was intrigued by your question and decided to contact Devcon directly. I was told by their applications engineer that the 5 minute epoxy will withstand DMF, acetone and most anything else. He said the only thing it doesn't stand up too well against is water!
He suggested soaking the sample in warm water for a few hours and then prying it apart with a razor blade. When I explained how fragile the sample was, he just said to soak it longer and it will come apart. If you still have trouble getting it apart, try contacting Devcon directly. They have a help section on their web site - unfortunately, I forgot to copy down their web site address, but I found it just by searching on "Devcon".
David Henriks TEL: 800-728-2233 (toll-free in USA) South Bay Technology, Inc. 714-492-2600 1120 Via Callejon FAX: 714-492-1499 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 Richard Beanland +44 1327 356363 } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello all. last week I stupidly stuck a Cu grid on top of the area of interest of a TEM cross section. Having tried to remove it for a few days, I thought I'd turn to the microscopy community for some help! The sample is Si, polished to less than ten microns thick (orangey colour), with Al and SiO2 on the top surface. It's the only one I have. I stuck the 2x1mm slot grid onto the sample with 5-minute epoxy (devcon); I didn't realise the grid was in the wrong place until a couple of hours later. Since then I've soaked it in dmf (dimethylformamide) [3 days] acetone [a few hours] and ashed it in nitrogen and oxygen [a couple of hours]. The sample is still in one piece and stuck to the grid. I've tried pushing it about with a fine hair but it's still well fixed. So, while it is soaking for a little longer in dmf and being ashed periodically, has anyone got any bright ideas how to rescue my sample?
Many thanks in advance,
Richard Beanland, Gmmt Ltd., Caswell, Towcester, Northants NN12 8EQ
I am planning to purchase a video or digital camera to document, analyze, and print images of paint and fiber samples using polarized light microscopy and fluorescence microscopy. Previous threads re video or digital cameras have focused largely on black and white images obtained using SEM or TEM and printed using high-DPI inkjets or dye-sub printers.
May I ask for recommendations or comments on purchasing a system (input through output) to deal with colored images of paint cross-sections, fibers, petrographic samples, etc.? This system would allow still images to be captured, analyzed/manipulated using image analysis software (including Photoshop), embedded in reports, and printed with near photographic quality in color.
I'll be pleased to provide more details, as requested.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Continuing the discussion of this topic. I used twin jet electropolishing a bit about 1970, but was frustrated by not being able to determine when a shiny surface was produced. (I had been able to see the sample in an old glass system operated manually). About that time, one of the early South Bay 550 polishers was ordered by me here at Argonne because it permitted magnified, IN SITU, viewing of the sample during polishing. This was needed to thin 1 or 2 new materials every week! The ease of use permitted accumulating reproducible data published in a 65 page report, (ANL-80-120), used world wide, a journal cover photo, and 12 other published articles. This work brought me the MSA "Technologist of the Year" award in 1994. About six 550 polishers are used exclusively at Argonne-some as long as 25 years! The unit can do jetting from one side, (for back-thinning to a special surface-lacquer protected), or both sides by inverting the sample after jet polishing about half way thru it. Microshield lacquer, (from South Bay), works great to protect surfaces from etching, dissolves in acetone, and may be thinned to reduce shrinkage when thinning soft, annealled copper for example. Also, the entire 3 m.m. disc surface can be polished via a 3 m.m. jet; using an external "timer/switch",and a D.C. power supply, planar "sectioning" of as little as 100 nm. can be removed from a surface. The jet polishing electrolyte and conditions will work. The large jet can be used to etch a surface for optical photos by simply reducing the "polishing" voltage about 20% for a couple of seconds! The 300 volt, 150 mA. capacity power supply exeeds other manufacturer's units and makes use of non-acid "BK-2" type electrolytes possible-a must for many materials. The line-of-sight optical shut off system can be fitted with a variety of color spectrum light sources for special uses. The standard infra red LED and detector bias may be independently adjusted to give the desired sensitivity setting. It will make electron transparant regions in pure annealled metal such as aluminum-with no hole! Of course the setting is normally set for a 20 micron hole with a very thin edge (quite reproducible, of course). Alignment of the parts is easy and stays set a long time. Even saphire light pipes are available for hydrofluoric acid or bromine/alcohol solutions. PVC plastic parts are available and may be substituted for metal ones for such strong chemical baths. Low temperatures of -50 degrees C. are no problem. The sample is accesible for rapid rinsig after swinging the detent-equipped jet support to one side. In 25 years of use, these instruments have saved one man per year in labor cost, (roughly $100,000/yr.), or $2,500,000--due to the ease and speed with which excellent samples can be made. About 90 to 95% of the samples attempted are good once- conditions are established. In my opinion, all the jet polishers have improved with time, but the South Bay 550 C and 550 D units are unmatched when it comes to working with the newer, difficult materials which should be viewd DURING thinning. They permit me to thin about 300 TEM foils/year in my spare time.
Might as well throw in my two cents worth on this topic. We generally make our projections slides from prints, rather than negatives. This, of course, requires making prints first, but since this is often done anyway, it's usually not a problem.
We use a seldom-mentioned film known as Kodak 5468, a direct positive film used, I think, as a motion picture stock. It's bright red and translucent---strange looking stuff. Our exposures on a 4-light copy stand range from 6-10 seconds at a lens opening of f/3.5 on a Canon 55mm macro lens, so it obviously requires a lot of light. Development is in Dektol diluted 1:1 with water. Yes, Dektol, the paper developer. Stop with water, fix with whatever you usually use and wash normally.
This film is cheap (still less than $25/100 ft., last I checked), development is easy, and the slides are quite good. The problem is that this film tends to undergo reduction-oxidation (redox) after a few years, giving a solarized appearance. To prevent this, use Kodak Brown Toner (TOXIC---use plenty of ventilation and gloves) diluted about 1:50 for a 30-second dip. This will often give your slides a slight brown tone. Some folks like this, some don't.
Not a perfect solution, but very useful for many purposes.
Randy Tindall Center for Electron Microscopy Southern Illinois University at Carbondale
Gerry et al., Perma Wash is advertised as an archival high speed wash for both film and paper that reduces wash time by 90%. It's made by Heico (a Cambrex Co.) along with other kinds of photographic chemicals. I've used it for years both as an EM specialist and as a photographer with no problems at all. You should check with local photographic darkroom supply house instead of any EM suppliers. Two suppliers that I know who do offer it are listed if you still can't find it in your area. For film Heico states archival permanence with one minute first water wash, one minute Perma Wash, one minute final water wash; for r.c. paper, it's two minutes, two minutes, two minutes; finally, for double weight papers, five minutes, five minutes, five minutes does the job. I hope this washes well for you. :-) :-) :-}
Camera World Crimson Tech PO Box 9426 325 Vassar Street 1809 Commonwealth Ave Cambridge, MA 02139 Charlotte, NC 28205 Pho: 800-868-3686 800-868-5150 704-375-8453 617-868-5150 Fax: 704-376-1826 617-499=4777
HEICO Chemicals Inc. Route 611 Delaware Water Gap, PA 18327
717-420-3900
Contains ammonium sulfite, sodium sulfite and water.
The container say it specifically removes silver sulfate.
Tom
Thomas Moninger moninger-at-emiris.iaf.uiowa.edu University of Iowa Central Microscopy Research Facility http://www.uiowa.edu/~cemrf Views expressed are mine.
} last week I stupidly stuck a Cu grid on top of the area of interest } of a TEM cross section. Having tried to remove it for a few days, I thought } I'd turn to the microscopy community for some help! } The sample is Si, polished to less than ten microns thick (orangey colour), } with Al and SiO2 on the top surface. It's the only one I have. I stuck the } 2x1mm slot grid onto the sample with 5-minute epoxy (devcon); I didn't real- } ise the grid was in the wrong place until a couple of hours later. Since } then I've soaked it in dmf (dimethylformamide) [3 days] acetone [a few } hours] and ashed it in nitrogen and oxygen [a couple of hours]. The sam- } ple is still in one piece and stuck to the grid. I've tried pushing it } about with a fine hair but it's still well fixed. } So, while it is soaking for a little longer in dmf and being ashed } periodically, has anyone got any bright ideas how to rescue my sample? } } Many thanks in advance, } } Richard Beanland,
} I was intrigued by your question and decided to contact Devcon directly. } I was told by their applications engineer that the 5 minute epoxy will with- } stand DMF, acetone and most anything else. He said the only thing it does- } n't stand up too well against is water! } He suggested soaking the sample in warm water for a few hours and then pry- } ing it apart with a razor blade. When I explained how fragile the sample } was, he just said to soak it longer and it will come apart.
} David } Dear Richard, Here's a thought: If just soaking won't do, you might try micro- waving the water-soaked specimen. That will heat up the water, but maybe not the Si, Al or Cu. I know metal is a no-no in a microwave oven, but there is so little that there shouldn't be a disaster here. I'd put ~50 ml of H2O in a beaker in the oven at the same time. Of course, I'd try it first with something other than the specimen. Good luck. Yours, Bill Tivol
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Continuing the discussion of this topic. I used twin jet electropolishing a bit about 1970, but was frustrated by not being able to determine when a shiny surface was produced. (I had been able to see the sample in an old glass system operated manually). About that time, one of the early South Bay 550 polishers was ordered by me here at Argonne because it permitted magnified, IN SITU, viewing of the sample during polishing. This was needed to thin 1 or 2 new materials every week! The ease of use permitted accumulating reproducible data published in a 65 page report, (ANL-80-120), used world wide, a journal cover photo, and 12 other published articles. This work brought me the MSA "Technologist of the Year" award in 1994. About six 550 polishers are used exclusively at Argonne-some as long as 25 years! The unit can do jetting from one side, (for back-thinning to a special surface-lacquer protected), or both sides by inverting the sample after jet polishing about half way thru it. Microshield lacquer, (from South Bay), works great to protect surfaces from etching, dissolves in acetone, and may be thinned to reduce shrinkage when thinning soft, annealled copper for example. Also, the entire 3 m.m. disc surface can be polished via a 3 m.m. jet; using an external "timer/switch",and a D.C. power supply, planar "sectioning" of as little as 100 nm. can be removed from a surface. The jet polishing electrolyte and conditions will work. The large jet can be used to etch a surface for optical photos by simply reducing the "polishing" voltage about 20% for a couple of seconds! The 300 volt, 150 mA. capacity power supply exeeds other manufacturer's units and makes use of non-acid "BK-2" type electrolytes possible-a must for many materials. The line-of-sight optical shut off system can be fitted with a variety of color spectrum light sources for special uses. The standard infra red LED and detector bias may be independently adjusted to give the desired sensitivity setting. It will make electron transparant regions in pure annealled metal such as aluminum-with no hole! Of course the setting is normally set for a 20 micron hole with a very thin edge (quite reproducible, of course). Alignment of the parts is easy and stays set a long time. Even saphire light pipes are available for hydrofluoric acid or bromine/alcohol solutions. PVC plastic parts are available and may be substituted for metal ones for such strong chemical baths. Low temperatures of -50 degrees C. are no problem. The sample is accesible for rapid rinsig after swinging the detent-equipped jet support to one side. In 25 years of use, these instruments have saved one man per year in labor cost, (roughly $100,000/yr.), or $2,500,000--due to the ease and speed with which excellent samples can be made. About 90 to 95% of the samples attempted are good once- conditions are established. In my opinion, all the jet polishers have improved with time, but the South Bay 550 C and 550 D units are unmatched when it comes to working with the newer, difficult materials which should be viewd DURING thinning. They permit me to thin about 300 TEM foils/year in my spare time.
We are having a mini-debate here over whether one can confidently say what side of the membrane an epitope on a membrane faces (e.g., towards the cytoplasm or lumen of the RER) based on the distribution of gold labeling. If most the labeling is on one side, would you be confident the epitope is solely on that side? One of my collaborators had a paper criticized by a reviewer who said this couldn't be done reliably.
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
OOPs--I intended to say that acetic acid may damage the aluminum film. It doesn't attack silicon from my experience.
best regards mark
Mark W. Lund, PhD Director } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"The state is good at simple tasks, like killing people and seizing their wealth. It has far more trouble reaching inside individuals and making them good." Doug Bandow
} The Company: } } Samsung Austin wants to offer you more than a job. We want to offer you the } chance to develop a career. Becoming involved in a fast-paced start-up } operation is both exciting and challenging and we look for employees who are } energetic, flexible, and team-oriented. } Are you willing to go the extra mile? Are you comfortable with change? Are } you interested in great benefits? Are you excited about working in a start-up } environment? Are you interested in a career where you will be an integral } part of a new company? Are you comfortable making decisions that will } influence the development of our corporate culture? } If the answer to these questions is "yes," then Samsung Austin is the company } for you. } For more information visit our web site at www.sas.samsung.com } } } The Position } } We are currently looking to hire an Entry Level Auger Technician. The } requirements for this position is simply an Associate Degree in a technical } major. Experience is preferred but not necessary. The position will involve } shift work. } } If someone know of a person who would consider this position or if you know } of a 2 year college that offers a course which covers Auger Electron } Spectroscopy please feel free to contact me. } } Resumes can be faxed or emailed to: } David A. Griffiths } Fax (512)491-1165 } Phone/VoiceMail(512)491-1403 } Pager (512)209-4132 } e-mail DGriffiths-at-sas.samsung.com
Richard Beanland described a problem involving a TEM grid stick to a sample in the wrong place, glued with a 5-Minute (Devcon) epoxy.
One is his comments was: ================================================ I've soaked it in dmf (dimethylformamide) [3 days] acetone [a few hours] and ashed it in nitrogen and oxygen [a couple of hours]. The sample is still in one piece and stuck to the grid. I've tried pushing it about with a fine hair but it's still well fixed. ================================================ Oxygen plasma etching, in an isotropic (I stress isotropic as opposed to anisotropic) plasma etcher should remove the epoxy. If you were correct in that you used "nitrogen and oxygen", then this is why it did not work. You have to use pure oxygen, no nitrogen. Even a small leak in the system, allowing just a small partial pressure of nitrogen will literally kill the etching rate. In air, literally nothing will happen.
Now, if in fact you were using pure oxygen, there could still be a leak problem, another reason why you did not get any etching. But the technique should work. Make sure the power is not more than 100 watts or else the sample might heat up to temperature that would not be acceptable.
Disclaimer: SPI manufactures an isotropic plasma etcher for doing this kind of etching of organic materials. You can see further information on our website as well as an explanation of the differences between isotropic vs. anisotripic etching.
Chuck
================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Does anyone have a tried and true method for the fixation and embedment of Tetrahymena? I would like to do some preembedding staining for nuclear proteins, and have been having trouble once the dehydration is started. The cells are blasted apart by the time I examine them under the electron microscope. The cells are intact after preembedding immunostaining (at least they appear to be under phase contrast). After a post immunostaining treatment with 1% glutaraldehyde, the cells are enrobed in agar (1:1 of 2% low-melting temperature agar) at 37C, dehydrated in a graded ethanol series (10, 30, 50, 75, 95, 100, 100 -- 15 minutes each) and then progressively infiltrated with LR White resin (all at room temp). Polymerization is at 55C for 24 hours.
Any help/ideas would be greatly appreciated.
Craig Lending Department of Biology SUNY Brockport Brockport, NY 14420 Phone: 716-395-5755 Fax: 716-395-2741 e-mail: clending-at-acs.brockport.edu
Damian - Sabatini (he made GA the EM fixative) did some experiments and declared that postfixation could be left for at least six months. I guess that is true for a few tissues. Lipids are not well fixed in GA and lipid rich tissues in particular suffer when post fixation is delayed. By how much - well how long is a piece of string? Postfix as soon as possible. Store in buffer refrigerated. What you do for SEM or LM matters very little, but for TEM this matters. Never store tissues in GA for TEM. GA crosslinks materials, overfixing with GA results in a coarser texture which obscures fine details at high resolution. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 400+ Links, MSDS ************************ http://www.proscitech.com.au } } 1. How long can I store rat tissue in glutaraldehyde fixative before } conpleting the tissue preparation process and not experience tissue } degradation? Can I go as long as three weeks? At this point, I don't } know if I will be processing for SEM (CPD) or TEM - depends on the LM } results. } } 2. Should I store in buffer rather than the fixative? Some other } solution? } } 3. Would I use a different formulation of fixative for this kind of
} delayed processing of tissue? } } So much for quick questions and thanks so much for your help! Now, I } have to go answer someone else's question. } } Damian Neuberger } neuberd-at-baxter.com
Dear Richard, The only way I have removed 5-minute epoxy from something was by gently heating. The epoxy softens at heat-gun temps (70 deg.C?). This was on a leaky air-pressure valve, not a grid, so I just heated it until I could peel it off.. The other suggestion would be acetone, at least overnight. If you ask a embedding/biologist EM type, they may know a solvent to dissolve epoxy. You wrote:
} Hello all. } last week I stupidly stuck a Cu grid on top of the area of interest } of a TEM cross section. Having tried to remove it for a few days, I thought } I'd turn to the microscopy community for some help! } The sample is Si, polished to less than ten microns thick (orangey colour), } with Al and SiO2 on the top surface. It's the only one I have. I stuck the } 2x1mm slot grid onto the sample with 5-minute epoxy (devcon); I didn't realise } the grid was in the wrong place until a couple of hours later. Since then } I've soaked it in dmf (dimethylformamide) [3 days] acetone [a few hours] and } ashed it in nitrogen and oxygen [a couple of hours]. The sample is still in } one piece and stuck to the grid. I've tried pushing it about with a fine hair } but it's still well fixed. } So, while it is soaking for a little longer in dmf and being ashed } periodically, has anyone got any bright ideas how to rescue my sample? } } Many thanks in advance, } } Richard Beanland,
Good day everyone I would appreciate your comments on what permanent mounting media are available which meet the correct refractive index of glass and do not lead to fading of toluidene blue-stained sections?
Thank you.
James Wesley-Smith EM Unit University of Natal Durban, South Africa
Kodak Rapid Process Copy film (if it is still available) or Kodak Direct MP film (is available, e.g. from SPI or Ted Pella, etc) are excellent for inexpensive single-process preparation of B & W transparencies from EM prints.
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa eurc-at-giraffe.ru.ac.za - tel: +27 461 318168 - fax: +27 461 24377
Here is one addition (hopefully valuable) to the EDX-Spot Mode discussion. =20 If your SEM has a specimen current meter, you can use that successfully for checking the beam position very accurately before and during the EDX analysis of small particles.=20
The technique is very simple. You first position the spot mode beam at high magnification, and then by moving the beam in X-Y direction you either maximize or minimize the specimen current reading depending on whether the average atomic number (=3D Z) of the particle is lighter or=20 heavier that that of the matrix. The great advantage in the use of the=20 specimen current (=3D absorbed electrons) for positioning the beam is that= =20 you same time maximize the X-ray emission from the particle and minimize the possible X-ray contribution from the matrix. This is due to the fact that absorbed electrons "sense" the shape of the particle under the specimen surface.
If the specimen current stays constant during the measurement of the EDX spectrum, then you can be absolutely certain that the beam did not leave the particle during the measurement. However, normally there is a small increase ( { 1 %) in the specimen current due to the contamination build-up.
=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4= =A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4= =A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4 =A4 =A4 =A4 Seppo J. Sivonen e-mail: seppo.sivonen-at-oulu.fi =A4=20 =A4 University of Oulu =A4=20 =A4 Institute of Electron Optics tel: +358-8-553 3140 =A4 =A4 Box 400 fax: +358-8-553 3149 =A4=20 =A4 FIN-90571 Oulu =A4 =A4 FINLAND http://koivu.oulu.fi/~eolwww/welcome.html =A4 =A4 =A4 =A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4= =A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4= =A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4=A4
I don't know about the refractive index, but I've used Epoxy resin as a mountant quite successfully. It doesn't seem to fade Toluidine Blue semi-thins.
DePeX is not too bad but I have had some fading over long periods.
---------- } From: James Wesley-Smith {wesleysm-at-biology.und.ac.za} } To: 'Microscopy-at-sparc5.microscopy.com' {Microscopy-at-Sparc5.Microscopy.Com} } Subject: LM coverslip mounting medium } Date: 17 July 1997 09:08 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Good day everyone } I would appreciate your comments on what permanent mounting media are } available which meet the correct refractive index of glass and do not lead } to fading of toluidene blue-stained sections? } } Thank you. } } } James Wesley-Smith } EM Unit } University of Natal } Durban, South Africa }
Perhaps I didn't make my original comments clear enough. But yes the unexposed negative goes in the enlarger film carrier in the enlarger. This has the big advantage that reduction is no problem. The disadvantage is that if your enlarger doesn't do one to one printing in the first place then you still can't do it - just the reductions. This is usually not a problem if you are doing this with e.m cut film because it is larger and so needs to be reduced. You could even do this with prints ie copying them from the baseboard of the enlarger onto the negative in the film carrier but the amounts of stray light needed for copying stand lights makes it more fiddly. I think someone else has already mentioned it is important to mask out stray light.
I stumbled upon this idea because in one lab, I worked, they had a large Durst Laborator 1000 enlarger with all the accessories. This allowed you to use it as a copy camera and gave you special large format film holders for the purpose. Another lab I worked in had a DeVere large format enlarger with full reduction facilities. It seemed reasonable therefore to use an ordinary Durst enlarger as a copy camera so that I could do the 'DeVere thing, in reverse. I just didn't have all of the clever light-tight film holders for masking unexposed film in the carrier but with care it works, anyway.
If I remember rightly many of the big Durst enlargers have a reversible mirror in the condensor system - our Durst 1200s look as if you can just take out the fitting so that it faces towards the operator rather than the lamp. You should then be able to view the image of your negative or whatever else is on the baseboard through the little window in the front of your enlarger and VOILA you have a simple camera which will do large format. I am sure it should be in the manual somewhere.
Bizarre it may seem, to use an enlarger to reduce, but isn't there a certain pleasing symmetry to it? Sorry to go on but I thought this might be of general interest.
DISCLAIMERS: I hasten to add you can only do this with some enlargers; it will disrupt normal printing in a one enlarger darkroom; you may damage the enlarger so be careful; if in doubt read the manual or ask the supplier/manufacturer; and I will refuse to re-imburse anyone for self-inflicted damage. I have no connections with Durst or De Vere other than as a satisfied user.
Malcolm Haswell ----------
Malcolm, This sounds bizarre. Are you talking about a contact print here on to film? Or do you mean leave the negative carrier in the carrier holder of the enlarger, and use it upside down? (sort of like using the enlarger as an upside down camera.)
Garry
} Garry } } we are drifting well away from the main theme, but you can still make slides } with your enlarger even if you haven't got close focussing on your lens. } It's more fiddly but if you have a light box you simply put the e.m. } negative on the light box under the enlarger and the film to be exposed in } the negative carrier. } The tricky bit is getting the position and focus right but you do most of } that by enlarging something of the right format first onto the light box. } Then of course when you're exposing the film in the enlarger you must } remember to turn on the light box and not the enlarger. } } I have used this a couple of times and it works fine in an emergency } although of course if you were doing a lot it would be easier to make the } slides with a close-up 35mm camera and a light box.. } } Malcolm Haswell } e.m. unit } University of Sunderland } UK } ---------- } From: Garry Burgess } To: 'Microscopy Society of America } Subject: RE: Rush Lab + Projection Slides } Date: 15 July 1997 18:12 } } {SNIP} } Neat method of making slides, by the way. I'm going to remember it for } possible future use. } } Yes, it works fine. But the biggest obstacle for someone doing this for the } first time is to make sure that they have a lens that is capable of making } such a small focused image. You also have to make sure that you use glass } slide holders, because negative film cannot stand up to the heat of a slide
} projector without glass protection, because it's not as tough as slide film. } } Garry
Tom I tend to agree with the reviewer. I have spent several years looking at the membrane cytroskeleton of muscle, and commonly the gold particles appear "extracellular" when we know for sure they are on the inside of the plasma membrane. The argument is that as an IgG is about 11nm long, tagged to a 5nm gold particle, and if an indirct tagging system is system is used this adds an another 11nm. this leads of a potential radius of 25nm from the label site. I am sure from quantitative studies it is less than this, though I am equally sure that the gold particle commonly does not reflect to epitope position exactly.
Simon
Tom Phillips wrote:
} ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } ----------------------------- } -----------------------------------------. } } We are having a mini-debate here over whether one can confidently say } what } side of the membrane an epitope on a membrane faces (e.g., towards the } } cytoplasm or lumen of the RER) based on the distribution of gold } labeling. } If most the labeling is on one side, would you be confident the } epitope is } solely on that side? One of my collaborators had a paper criticized } by a } reviewer who said this couldn't be done reliably. } } Thomas E. Phillips, Ph.D. } Associate Professor of Biological Sciences } Director, Molecular Cytology Core Facility } 3 Tucker Hall } University of Missouri } Columbia, MO 65211 } (573)-882-4712 (voice) } (573)-882-0123 (fax)
-- Simon C. Watkins Ph.D. Associate Professor Director CBI University of Pittsburgh Pittsburgh PA 15261 tel:412-648-3051 Fax:412-648-2004 URL:http://sbic6.sbic.pitt.edu
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Reply to: RE} Au labeling of membranes - can you judge sideness?
Dear Thomas, Your discussion regarding which side of the membrane the labeling is on should include how to determine the resolution of the labeling technique. The resolution is determined by the size of the probe used for visualization as well as the antibody used. For example, an IgG molecule has an extended length of about 8-10nm and a 5nm gold particle would be expected to have a total diameter of 7-8nm of covered with protein-A. The resolution comes from the circular radius of this complex from the antigen outward and upwards. The antibody could fall anywhere within this radius, thus making it difficult to say if it is on the inside or outside of the membrane. You could confidently say it labels the membrane but to go further may be stretching what you see visually. Section thickness also plays a part in reducing the resolution. For further information you may want to pick up a book by Garreth Griffiths called "Fine structure immunocytochemistry" printed by Springer-Verlag ISBN 3-540-54805-X. It is a comprehensive look at a number of variables pertaining to immunocytochemistry.
Linda Chicoine Center for Cell Imaging Dept. of Cell Biology Yale University 203-785-3646 phone 203-785-7226 fax
--------------------------------------
We are having a mini-debate here over whether one can confidently say what side of the membrane an epitope on a membrane faces (e.g., towards the cytoplasm or lumen of the RER) based on the distribution of gold labeling. If most the labeling is on one side, would you be confident the epitope is solely on that side? One of my collaborators had a paper criticized by a reviewer who said this couldn't be done reliably.
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
------------------ RFC822 Header Follows ------------------ Received: by QuickMail.Yale.edu with SMTP;16 Jul 1997 18:41:02 -0400 Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id RAA05780 for dist-Microscopy; Wed, 16 Jul 1997 17:10:11 -0500 Received: from biosci.mbp.missouri.edu (biosci.mbp.missouri.edu [128.206.15.250]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id RAA05777 for {Microscopy-at-msa.microscopy.com} ; Wed, 16 Jul 1997 17:10:09 -0500 Received: from [128.206.15.189] by biosci.mbp.missouri.edu with ESMTP (Eudora Internet Mail Server 1.1.2); Wed, 16 Jul 1997 17:21:38 -0500 Message-Id: {v03007801aff2f8f44010-at-[128.206.15.189]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Could someone please tell me how to unsubscribe to the Microscopy List Server? I plan to be a way for the next couple of weeks and would like to turn the flood of messages off. Thanks Rob Willson Dept of Anatomy and Cell Biology Tufts University
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Tom Phillips wrote: ....."what side of the membrane an epitope on a membrane faces (e.g., towards thecytoplasm or lumen of the RER) based on the distribution of gold labeling."....
I would also agree that the normal gold label may be to large, because of the size of the coupled IgG moelcule. What about the Nanogold labels: 1.4 nm gold attched to FAB fragment? I don't know what the total size would be, but much smalelr than if coupled to igG.Would this be small enough? the silver enhancement, to enlarge the size for viewing on TEM, is done after label. Louisa Howard EM Facility, 6044 Gilman Dartmouth College Hanover, NH, 03755
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Well, i dont know if this is a dumb solution, but you could try to etch the copper away with somehting like dilute nitric acid. i dont know the dangers with silicon or the aluminum, but perhaps you could find an etchant that would attack only the copper.
Michael T. Marshall Research Engineer, Electron Microscopy University of Illinois at Urbana-Champaign Frederick Seitz Materials Research Laboratory 104 South Goodwin avenue Urbana, IL 61801-2985 (217) 244-8193 fax: (217) 244-2278
For what it's worth regarding the use of Permawash: I have negatives that I washed according to their (Permawash) instructions at least 25 years ago and they are still in perfectly good condition.
Thanks to all who replied to my question about storing rat tissue in fixative. Here is a cut and paste summary of replies from all over the world! for anyone who is interested.
Sabatini (he made GA the EM fixative) did some experiments and declared that postfixation could be left for at least six months. I guess that is true for a few tissues. Lipids are not well fixed in GA and lipid rich tissues in particular suffer when post fixation is delayed. By how much - well how long is a piece of string? Postfix as soon as possible. Store in buffer refrigerated. What you do for SEM or LM matters very little, but for TEM this matters. Never store tissues in GA for TEM. GA crosslinks materials, overfixing with GA results in a coarser texture which obscures fine details at high resolution.
I have stored samples in 2% glut. in 0.1M sodium cacodylate for 2 weeks because I forgot about them. These were cell cultures and they turned out OK, I embedded these into epon/araldite. I have also stored things in buffer after fixation for a few weeks and this turned out OK too.
Dysktra's book on biological EM has micrographs of mouse kidney stored in formaldehyde/glut fix for something like 2 years that looks pretty good.
I often have kept things in GA for as long as 3 weeks, provided that it is kept cold. (There may be some problem with microtubule disassembly, according to some). Over time, the GA will break down 3weeks).
Store in buffer: No. You stand the risk of fixative being washed out and the possibility of "de-fixing" the material. You also could encourage bacterial growth eventually.
My recommendation would be to fix, wash, and post-fix in OsO4. If you then dehydrate to 70% EtOH or Acetone,it will keep for YEARS!
Although we try not to store anything, we prefer to store our samples in Trumps fix. A buffered glute/form combo. We have stored for weeks at a time and had good results
There is some evidence to suggest that storage in Trumps or half Karnovsky rather than pure glut (i.e. glut plut some paraformaldehyde)
I have stored tissue in glutaraldehyde or Trump's fix(glut/para fix) for years before using it for TEM. I wouldn't store it in buffer.
I would probably do a normal fix and then switch to buffer and store cold. I don't believe you will see any degradation as long as you stay away from post-fixation with osmium before storage; I would leave that go until you were ready to proceed with prep.
Damian- I've always tried to avoid any long term storage in fix, however storing samples after fixation in a 0.2M buffer has seemed to work well for up to 1-2 weeks. And after reading the thread on lipid preservation, I think it wise to osmicate prior to the storage in buffer too. _mike
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } Good day everyone } } I would appreciate your comments on what permanent mounting media are } } available which meet the correct refractive index of glass and do not } lead } } to fading of toluidene blue-stained sections? } } I learned a trick from a DuPont-Sorvall rep (that tells you how long ago that was!) that retards or eliminates ALL fading caused by oxidants in the mounting medium. Add 1-2% BHT (the preservative used in bologna, hot dogs, etc.) to any mounting medium. There should be a bottle of the stuff sitting on the shelf in the EML at U.C. Berkeley that has enough to supply every EM lab in the country...
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } Good day everyone } } I would appreciate your comments on what permanent mounting media are } } available which meet the correct refractive index of glass and do not } lead } } to fading of toluidene blue-stained sections? } } I learned a trick from a DuPont-Sorvall rep (that tells you how long ago that was!) that retards or eliminates ALL fading caused by oxidants in the mounting medium. Add 1-2% BHT (the preservative used in bologna, hot dogs, etc.) to any mounting medium. There should be a bottle of the stuff sitting on the shelf in the EML at U.C. Berkeley that has enough to supply every EM lab in the country...
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
Richard Beanland +44 1327 356363 wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello all. } last week I stupidly stuck a Cu grid on top of the area of interest } of a TEM cross section. Having tried to remove it for a few days, I thought } I'd turn to the microscopy community for some help! } The sample is Si, polished to less than ten microns thick (orangey colour), } with Al and SiO2 on the top surface. It's the only one I have. I stuck the } 2x1mm slot grid onto the sample with 5-minute epoxy (devcon); I didn't realise } the grid was in the wrong place until a couple of hours later. Since then } I've soaked it in dmf (dimethylformamide) [3 days] acetone [a few hours] and } ashed it in nitrogen and oxygen [a couple of hours]. The sample is still in } one piece and stuck to the grid. I've tried pushing it about with a fine hair } but it's still well fixed. } So, while it is soaking for a little longer in dmf and being ashed } periodically, has anyone got any bright ideas how to rescue my sample? } } Many thanks in advance, } } Richard Beanland, } Gmmt Ltd., } Caswell, } Towcester, } Northants NN12 8EQ } } Tel +44 1327 356363 } Fax +44 1327 356775 } e-mail richard.beanland-at-gecm.com Dear Mr. Beanland,
We have available an Epoxy Dissolver that is supposed to work and all 2 component epoxies. As I have not tried this product on all epoxies available, I do not know if it will work with this particular type.
If you can, visit your local pharmacy and ask them for some DMSO(Dimethylsulfoxide). This is the primary ingredient and it will need to be heated to operate effectively.
Please let me know if you have any other questions.
Sincerely,
Gary Liechty Allied High Tech Products, Inc. 2376 E. Pacifica Pl. Rancho Dominguez, Ca. 90220 310-635-2466 800-675-1118 310-762-6808 Fax
Products for Materialographic, SEM and TEM sample preparation
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From: Bernie Kestel-Argonne National Lab
I forgot to mention an important feature on the South Bay 550 series jet polishers in my previous message. The "open faced" specimen retainer has only a 0.0015" thick polyethylene diaphram with a central hole in it to hold the specimen down on a platinum tipped holder. This thin sheet offers almost NO flow resistance or bubble trapping area near the specimen. The all important electrolyte viscosity/polishing film can be thicker with this design, producing smoother finished surfaces while "bridging" across grain boundaries, precipitates and other features. That is why I do most polishing at -45 C. or so and add butyl cellosolve to increase electrolyte viscosity to 10-12 centipoises-ideal. (Like half & half from a refrigerator, approx.). Of course a PVC cap with a central hole positions the specimen laterally. I pass this along to hopefully ease someones prep. burden - -I'm NOT financially connected to South Bay! I feel this unit is like driving a modern auto compared to a hand cranked, manual shifted, no air conditioning machine. Take the easy route!
Our lab has been using Glutaraldehyde that expired in November of 1995, but has been kept refrigerated since then. Lately we have been noticing problems with our fixation, and I was wondering if perhaps it is because of this old Glutaraldehyde. Does anyone know how important these expiry dates really are with respect to its effect on fixation?????????
I don't mean to be hypercritical but doesn't everybody else see how to unsubscribe at the top of their message. I am a new subscriber (one week) and there must have been at least 3 messages similar to this one. Read the rules or does the old adage apply that:
OLD MICROSCOPISTS NEVER DIE THEY JUST LOSE THEIR RESOLUTION.
Mike Mead
RWILLSON-at-pearl.tufts.edu wrote:
} ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } ----------------------------- } -----------------------------------------. } } Could someone please tell me how to unsubscribe to the Microscopy } List Server? I plan to be a way for the next couple of weeks and } would like } to turn the flood of messages off. } Thanks } Rob Willson } Dept of Anatomy and Cell Biology } Tufts University
{P} I don't mean to be hypercritical but doesn't everybody else see how to unsubscribe at the top of their message. I am a new subscriber (one week) and there must have been at least 3 messages similar to this one. Read the rules or does the old adage apply that:
{P} {FONT SIZE=+2} OLD MICROSCOPISTS NEVER DIE {/FONT} {BR} {FONT SIZE=+2} THEY JUST LOSE THEIR RESOLUTION. {/FONT} {FONT SIZE=+2} {/FONT}
{P} RWILLSON-at-pearl.tufts.edu wrote: {BLOCKQUOTE TYPE=CITE} ------------------------------------------------------------------------ {BR} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
{P} Could someone please tell me how to unsubscribe to the Microscopy {BR} List Server? I plan to be a way for the next couple of weeks and would like {BR} to turn the flood of messages off. {BR} Thanks {BR} Rob Willson {BR} Dept of Anatomy and Cell Biology {BR} Tufts University {/BLOCKQUOTE} {/HTML}
After reading the summary Damian posted I wanted to chip in my 2 cents worth.
One respondent recommeded storage in fix since long-term buffer storage might "unfix" the specimen. I find it hard to believe that buffer could undo the powerful crosslinking glut causes. I have heard of this possibility with formalin fixation but that is not nearly as powerful a fix as glut. Anyone know of any experiments in this area?
Another responsent advocated storage in ethanol or acetone after post-fixing in OsO4, stating the tissue would keep for years. It will keep but a lot of cytoplasm will be extracted. Hayat's book, Prin. and Tech. of EM (1981 edition) discusses this and gives references and an illustration on pp 150-155.
I would store in buffer, changing it often if I could not complete processing soon.
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 e-mail: mcauliff-at-umdnj.edu ***************************************************************
I am in the market for a used clinical ultracentrifuge as well as an oven to be used during TEM specimen embedding. If anyone knows of a lab that is interested in selling these items, please contact me directly.
Thanks,
Dan Caruso c/o Eugene Gordon Biological Technician Medjet, Inc. 1090 King Georges Post Road Edison, NJ 08837 Phone: (732) 738-3990 Fax: (732) 738-3984 MEDJET-at-WORLDNET.ATT.NET
Garry Burgess wrote: ================================================== Our lab has been using Glutaraldehyde that expired in November of 1995, but has been kept refrigerated since then. Lately we have been noticing problems with our fixation, and I was wondering if perhaps it is because of this old Glutaraldehyde. Does anyone know how important these expiry dates really are with respect to its effect on fixation?????? ================================================== You are quite correct in that there can be some degree of arbitrariness in the statement of the expiration date. At least to us, the expiration date should correlate with some future point in time, after which, one could expect to see some deterioration of performance. Of course many of us know that film and paper, if properly stored don't turn into pumpkins on their expiration dates.
In the case of glutaraldehyde, the two most important factors influencing what will be the actual expiration date (as opposed to that stamped on the product), in the case of glut would be
a) starting purity of the ampouled product, since it is an autocatalytic reaction, and once the dimers and trimers reach some critical level, deterioration (e.g. polymerization) can proceed quite quickly. Hence a starting purity of the least amounts of the dimers and trimers, etc. relative to a glut with higher levels, would be expected to have longer shelf life. But we are talking about variations in starting purities that, when fresh, I would expect, would give any user good results.
b) thermal history during shipment. It is quite an education to follow the progress of a shipment and to see to what levels of heat exposure a particular shipment is exposed. Over the years I have myself "visited" UPS and FedEx trucks and have been quite surprised at how hot the inside of a truck can get on a hot summer day. Indeed some of the large warehouse type sorting rooms of the courier services are not air conditioned. Travelling on a highway for hours at a time with a hot summer sun beating down on the trailer leads to sometimes very hot temperatures. I have also been in institutional receiving departments that have been like a furnace in the middle of the summer. And I have been in receiving departments during the winter, and on the coldest of days, when supplemental heat is being provided by portable space heaters and the temperature of nearby boxes seem almost too hot to touch (well a slight exaggeration, because they were not breaking out into flames, but you get my point).
You would not want to know what a receiving department is like at Cairo University in June and July, where the temperatures are over 100 deg. in the shade!
So the answer is, you don't know what has been that thermal history. Have you been lucky or have you been unlucky? Most people would not want to leave the outcome of their important experiments to the chance that it was OK.
So at least in our case, and I suspect others too, supplier applied expiration dates are a best estimate to predict when one should start thinking about ordering fresh material and tossing the old material.
Chuck
PS: In the specific case at hand, if there is even the slightest trace of a precipitate in the ampoule, just write it off and don't even try using it. But in this case, even if there was not a precipitate, that fact we are approaching two years beyond the expiration date (e.g. 11/95), it would be good cause the do the same thing.
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I don't have any experience with using it on Devcon 5 minute epoxy, but I have found that DMSO will dissove several epoxies quite well. Observe the usual precautions about DMSO, i.e. keep it off your hands, especially when it may contain any toxic substance.
Sincerely, Andy Andy Buechele The Catholic University of America 409 Hannan Hall Washington, D.C. 20064 (202) 319-4995 FAX: (202) 319-4469
I don't have any experience with using it on Devcon 5 minute epoxy, but I have found that DMSO will dissove several epoxies quite well. Observe the usual precautions about DMSO, i.e. keep it off your hands, especially when it may contain any toxic substance.
Sincerely, Andy
Andy Buechele The Catholic University of America 409 Hannan Hall Washington, D.C. 20064 (202) 319-4995 FAX: (202) 319-4469
George & www: These things were published over twenty years ago. What concerns me about our microscopy forum is that many of us are re-iterating believes or quoting from memory. Looking things up is considerable work and books do not include all important knowledge previously published. Here is a bit of my memory: I recall a discussion between Sjostrand and Cosslett almost 30 years ago. Sjostrand had published biological sections and claimed 10 A resolution and told the group that he was going to reduce that substantially. Cosslett then got up and with a few figures proved (?) that 10 A was as well as could be done with fixed tissues in sections. Later there were publications showing that in monolayers, cells required only three minutes of 1% (?) GA fixation. Beyond that, over fixation caused artefact - meaningless granularity, which obscured the finest details. Nothing to affect the average x30k micrograph, but at 300k its a big factor. Over fixing is not a good practise. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 400+ Links, MSDS ************************ http://www.proscitech.com.au
---------- } From: George C. Ruben {George.C.Ruben-at-Dartmouth.EDU} } To: jim-at-proscitech.com.au } Subject: re: storing in fixative } Date: Thursday, 17 July 1997 21:39 } } --- You wrote: } Never store tissues in GA for TEM. GA crosslinks materials, overfixing with } GA results in a coarser texture which obscures fine details at high } resolution. } --- end of quote --- } } What embedding resolution do we we have if fixation is done correctly and does } overfixing effect resolution? Have you run standards or are you just talking } about the qualitative details in a picture after post fixing and staining-- } } --George C. Ruben } Dept . Biological Sciences
Could anyone tell me what to call a lens that is located between a galvano-scanner and objective lens in the light path of a scanning laser microscope?
I'm translating a manual of a scanning laser microscope from Japanese into English. (Is it *scanning laser* microscope or *laser scanning* microscope anyway?) And I can't find a proper English term for this lens which is called "pupil projection lens" whe n translated literally.
Any suggestion is welcomed.
Thanks in advance.
Chiba Atsushi [(Mr.) -- *Chiba* is my surname] Voice: (+81) 010-045-9451
Dear Microscopists, we plann to use TEM Philips 420 for Low-dose work. We will need Low-dose system for this microscope. Unfortunately Philips said us,that they didn't produce Low-dose for this kind of microscope,already. Can you help us please, how or where we could get this Low-dose unit or how to work in low-dose conditions /to prevent destroing samples during focusing/ without this system on TEM Philips 420 ? All your responces or opinions will wery useful for us.
Thank You very much Milos
Milos Motejl Lab. of Biomembranes South Bohemian University Ceske Budejovice Czech Republic
I tested EDTA versus Chromium Potossium Sulphate and found the latter much better for ultrastructural preservation (I was looking at bone-lining-cells) as long as the pieces of bone were tiny and the decalc short (few days) I have the original reference somewhere around if a medline search doesn't do the trick. Dunno about ascorbic acid as I didn't try that.
Amanda
Miss A.J.Wilson Electron Microscope Unit St George's Hospital Medical School Cranmer Terrace Tooting London SW17 ORE Tel: 0181 725 5220 awilson-at-sghms.ac.uk awilson-at-aw.u-net.com
We have two LKB 7800 series Knifemakers that still render outstanding service. Please have a look at our publication "further modification of the LKB 7800 series Knifemaker for improved reproducibility in breaking'cryo' knives.(ref Jnl of Microscopy Vol. 168, Pt 1. Nov 1992 pp 111 - 114). The simple modifications suggested in this paper transformed our knifemaker's such that any thought of replacing them with newer 'better' units vaporised. I remember that Jan Slot, on a visit to our lab, was very impressed with the modification and performance of the knifemakers some years back
Tony Bruton University of Natal Pietermaritzburg South Africa
} } } Linda Fox {lfox1-at-wpo.it.luc.edu} 2/July/1997 06:57pm } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Does anyone know where we can get our LKB 7800B knife breaker serviced? If possible, by someone in the Chicago area? It is making sporadically bad knives and we have adjusted all the knobs by the instruction booklet. If it needs to be replaced, can anyone recommend a good one? This one has been with us for over 20 years.
Thanks, Linda Fox, Loyola Univ. Medical Center, Chicago lfox1-at-wpo.it.luc.edu
Has anyone out there had first hand experience using the freeze fracture attachment for an Edwards 306 vacuum coating unit? Please reply directly to me.
Thank you very much.
James Wesley-Smith Electron Microscope Unit University of Natal Durban, South Africa
First of all, I'd like to thank all of those who cared to respond to my question, especially Dr. Garber from SPI who gave a detailed and thoughtful response. And OK, I get the message, it's just not worth the risk.
But I have another question that perhaps you people also might have some thoughts on. If I am understanding them properly, some of the Pathologists here claim that they can tell the difference between a specimen that was delayed before putting into Glut., resulting in artifact such as swollen mitochondria, vs. poor fixation as a result of outdated glutaraldehyde, such as damaged membranes in general. Is this sort of reasoning valid?
I agree with Dr. Garber, there is a good possibility the fix has gone off. If I were you, I would order a small replacement supply and test this to make sure your problem is the fix- that way you assure yourself of not wasting your supply. Order more and throw out the old glut. once your certain it's the problem.
My two cents,
Karen Pawlowski Lab. Tech. UT Southwestern Medical Center, PhD Student UT Dallas, Dallas TX
Are you by chance the Jim Martin I know with MA State Police?
I have been working this year to help bring the new Polaroid "DMC" Digital Microscope Camera to market. If we can confirm your location, I would recommend arranging a demo of this camera which should be EXCELLENT for all but the very lowest-level fluorescence work. Also, we will be featuring the DMC at the IAI meeting at Danvers, MA if you're going.
You'll need a WIN-95/Pentium system (Mac version drivers on the way in about 30-days). Recommend at least 32mb RAM and a hard drive large enough to handle your library of images (1.3mb or 5.5mb uncompressed, depending on resolution selected). The system plugs-in directly to any software that is TWAIN compatible (that's almost anything!).
Printers? Anything you want. Dye-sub is generally the best quality, but I have had outstanding success on the ink jet printers (like HP 870, et. al.) if the best media are used. Try the "premium glossy" papers, or even the "premium" clay-coated (matte finish) papers.
Get back to me with questions: corlb-at-polaroid.com Also there are specifications on the Polaroid Web Page: www.polaroid.com, look under "Polaroid at work".
______________________________ Reply Separator _________________________________ Subject: forensic/materials science, video-digital imaging Author: John D Warren at ~575ts2 Date: 7/16/97 6:35 PM
---------------------------------------------------------------------- -- The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com ---------------------------------------------------------------------- -.
I am planning to purchase a video or digital camera to document, analyze, and print images of paint and fiber samples using polarized light microscopy and fluorescence microscopy. Previous threads re video or digital cameras have focused largely on black and white images obtained using SEM or TEM and printed using high-DPI inkjets or dye-sub printers.
May I ask for recommendations or comments on purchasing a system (input through output) to deal with colored images of paint cross-sections, fibers, petrographic samples, etc.? This system would allow still images to be captured, analyzed/manipulated using image analysis software (including Photoshop), embedded in reports, and printed with near photographic quality in color.
I'll be pleased to provide more details, as requested.
First of all, I'd like to thank all of those who cared to respond to my question, especially Dr. Garber from SPI who gave a detailed and thoughtful response. And OK, I get the message, it's just not worth the risk.
But I have another question that perhaps you people also might have some thoughts on. If I am understanding them properly, some of the Pathologists here claim that they can tell the difference between a specimen that was delayed before putting into Glut., resulting in artifact such as swollen mitochondria, vs. poor fixation as a result of outdated glutaraldehyde, such as damaged membranes in general. Is this sort of reasoning valid?
We have two grades of glutaraldehyde - EM & BIO, and I believe you are talking about the former.
We have done some study on storage and the polymer peak at 230nm, and found material in a sealed ampule staying alright for up to two years. But since the customer will be breaking open an ampule, take out to lab, withdraw a little, and then put it back in the refrigerator, we figured these "in's and out's" will cut the stability to some extent and put an expiry date of one year.
Assuming that your lot with an expiration of November, 1995, was not taken out and then put back into the refrig too frequently, it could be stable for at least six more months; i.e., June, 1996, if not until the end of 1996. But now it is well beyond that period and so has possibly some polymer which causes it to be less effective in fixation. You should probably consider buying a new lot.
Good luck!
DR. PARASARAN POLYSCIENCES, INC.
---------- } From: Garry Burgess {GBurgess-at-exchange.hsc.mb.ca} } To: 'Microscopy Society of America - Mailing List' {microscopy-at-sparc5.microscopy.com} } Subject: Expired Glutaraldehyde } Date: Thursday, July 17, 1997 12:40 PM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Our lab has been using Glutaraldehyde that expired in November of 1995, } but has been kept refrigerated since then. Lately we have been noticing } problems with our fixation, and I was wondering if perhaps it is because } of this old Glutaraldehyde. Does anyone know how important these expiry } dates really are with respect to its effect on fixation????????? } } Not properly fixed, } } Garry
Along the same lines, we use the brightness of the video signal to make sure we are on the feature of interest. Our JEOL 840A has LED meters for contrast and brightness that we can monitor even in spot mode. We even used to do this on our JEOL U3, but it has been gone so long now I cannot remember exactly how we watched for it.
At 12:27 PM 7/17/97 +0300, you wrote: } Here is one addition (hopefully valuable) to the EDX-Spot Mode discussion. } } If your SEM has a specimen current meter, you can use that successfully } for checking the beam position very accurately before and during the EDX } analysis of small particles. } } The technique is very simple. You first position the spot mode beam } at high magnification, and then by moving the beam in X-Y direction } you either maximize or minimize the specimen current reading depending on } whether the average atomic number (= Z) of the particle is lighter or } heavier that that of the matrix. The great advantage in the use of the } specimen current (= absorbed electrons) for positioning the beam is that } you same time maximize the X-ray emission from the particle and minimize } the possible X-ray contribution from the matrix. This is due to the fact } that absorbed electrons "sense" the shape of the particle under the specimen } surface. } } If the specimen current stays constant during the measurement of the EDX } spectrum, then you can be absolutely certain that the beam did not leave } the particle during the measurement. However, normally there is a small } increase ( { 1 %) in the specimen current due to the contamination build-up. ---------------------------------------------------- Warren E. Straszheim 270 Metals Development, Ames Lab/ISU, Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-3091
--IMA.Boundary.996252968 Content-Type: text/plain; charset=US-ASCII Content-Transfer-Encoding: 7bit Content-Description: cc:Mail note part
Nora,
We examine similar samples in our lab on occasion. I would need more information on your samples to give a more specific procedure, but here are a few tips:
a) pigments in mineral oil: try diluting sample about 1:20 in hexane, heptane, or mineral spirits (i.e., a compatible solvent), let pigments settle out by gravity overnight (if they will), otherwise spin down gently in a centrifuge. Decant off the supernatant (i.e., the mineral oil in the solvent) without losing the pigments. Add more solvent to the container and try to redisperse the pigments. With some method development, you should be able to obtain a dispersion of the pigment in the solvent with only a little of the mineral oil left. Then put a droplet of this dispersion on a suitable polished substrate (carbon?), and wick off a little of the solvent with a kimwipe.
b) pigments in an oil/water emulsion. If you mean that the sample is an oil in water emulsion like a latex, dilute the sample in water 1:20, put a droplet on the substrate, and wick off the water. In this way you should be able to separate the 'oil' and the pigments enough to pick out the pigments and image or analyze them.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
, Dear friends from the list,
We have just received an enquiry about SEM analysis for: a) pigments particles dispersed in mineral oils b) pigments particles in oil-water emulsions
Does any of you have any experience in that subject? Unfortunately, we do not count with equipment for sample preparation other than the sputter-coater and carbon evaporator.
Any help will be very welcome.
Thanks in advance,
Nora Pratta Centro Regional de Investigacion y Desarrollo Santa Fe - Argentina
Received: from host68.sherwin.com (148.141.1.67) by ccgate.sherwin.com with SMTP (IMA Internet Exchange 2.1 Enterprise) id 00020513; Mon, 14 Jul 97 14:30:39 -0400 Received: from sparc5.microscopy.com by host67.sherwin.com with SMTP id AA08484 (InterLock SMTP Gateway 3.0 for {dfklimovich-at-sherwin.com} ); Mon, 14 Jul 1997 14:25:28 -0400 Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id JAA26094 for dist-Microscopy; Mon, 14 Jul 1997 09:27:45 -0500 Received: from alpha (alpha.arcride.edu.ar [200.9.237.9]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id JAA26091 for {MICROSCOPY-at-msa.microscopy.com} ; Mon, 14 Jul 1997 09:27:39 -0500
Chiba Atsushi wrote: :Could anyone tell me what to call a lens that is located between a galvano-scanner and objective lens in the light path of a scanning laser microscope? : :I'm translating a manual of a scanning laser microscope from Japanese into English. (Is it *scanning laser* microscope or *laser scanning* microscope anyway?) And I can't find a proper English term for this lens which is called "pupil projection lens" wh e : :Any suggestion is welcomed. : :Thanks in advance.
There are several words that can be used, but "transfer lens" is very common. Also used are "telecentric lens" and "relay lens."
best regards mark
Mark W. Lund, PhD Director } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"The state is good at simple tasks, like killing people and seizing their wealth. It has far more trouble reaching inside individuals and making them good." Doug Bandow
In response to Garry Burgess's question regarding swollen mitochondria sometimes referred to as "popcorn" mitochondria, the only time I have seen them perfectly preserved was after whole body perfusion of fixative through the heart.
It is likely that pathologists do not have tissue preserved in this way from diagnosing human diseases. I guess for argument sake, Garry, if I were in your shoes I would ask the pathologist (tactfully) weather the swollen mitochondria could be the result of the pathology and not poor fixation.
For perspective, though, I should tell you that I once worked in the Anatomy Department at Loma Linda University and the professor there didn't believe in purified glutaraldehye. He used the 25% stuff and kept it under the lab bench. His belief was that the older it got the better....something to do with the ratio of dimer to trimer. If you want to check his publications, I believe his name was Robert Schultz.
{HTML} {FONT SIZE=+1} In response to Garry Burgess's question regarding swollen mitochondria sometimes referred to as "popcorn" mitochondria, the only time I have seen them perfectly preserved was after whole body perfusion of fixative through the heart. {/FONT} {FONT SIZE=+1} {/FONT}
{P} {FONT SIZE=+1} It is likely that pathologists do not have tissue preserved in this way from diagnosing human diseases. I guess for argument sake, Garry, if I were in your shoes I would ask the pathologist (tactfully) weather the swollen mitochondria could be the result of the pathology and not poor fixation. {/FONT} {FONT SIZE=+1} {/FONT}
{P} {FONT SIZE=+1} For perspective, though, I should tell you that I once worked in the Anatomy Department at Loma Linda University and the professor there didn't believe in purified glutaraldehye. He used the 25% stuff and kept it under the lab bench. His belief was that the older it got the better....something to do with the ratio of dimer to trimer. If you want to check his publications, I believe his name was Robert Schultz. {/FONT} {FONT SIZE=+1} {/FONT}
In response to Garry Burgess's question regarding swollen mitochondria sometimes referred to as "popcorn" mitochondria, the only time I have seen them perfectly preserved was after whole body perfusion of fixative through a living and pumping heart.
Obviously, it is not likely that pathologists have tissue preserved in this way from living humans. I guess for argument sake, I would ask the pathologist weather the swollen mitochondria and other membrane defects could be the
result of the pathology and not poor fixation.
On the other hand, dead cells such as cuticle and cortical cells in hair have well defined cell membrane complexes that look well preserved even if you don't fix the hair--although cell organelles like mitochondria are missing.
For perspective, I should tell you that I once worked in the Anatomy Department at Loma Linda University and the professor there didn't believe in purified glutaraldehye. He used the 25% stuff and kept it under the lab bench. His belief was that the older it got the better....something to do with the ratio of dimer to trimer. I'm not a chemist, so I don't know. I'm sure the Ph.D.'s at the companies who sell the "good" stuff have their point of view.
} ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } ----------------------------- } -----------------------------------------. } } First of all, I'd like to thank all of those who cared to respond to } my } question, especially Dr. Garber from SPI who gave a detailed and } thoughtful response. And OK, I get the message, it's just not worth } the } risk. } } But I have another question that perhaps you people also might have } some } thoughts on. If I am understanding them properly, some of the } Pathologists here claim that they can tell the difference between a } specimen that was delayed before putting into Glut., resulting in } artifact such as swollen mitochondria, vs. poor fixation as a result } of } outdated glutaraldehyde, such as damaged membranes in general. Is } this } sort of reasoning valid? } } Just curious, } Garry
{HTML} {FONT SIZE=+1} ----------------------------------------------------------------------- {/FONT} {BR} {FONT SIZE=+1} The Microscopy ListServer -- Sponsor: The Microscopy Society of America {/FONT} {BR} {FONT SIZE=+1} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com {/FONT} {BR} {FONT SIZE=+1} -----------------------------------------------------------------------. {/FONT} {FONT SIZE=+1} {/FONT}
{P} {FONT SIZE=+1} In response to Garry Burgess's question regarding swollen mitochondria {/FONT} {BR} {FONT SIZE=+1} sometimes referred to as "popcorn" mitochondria, the only time I have seen {/FONT} {BR} {FONT SIZE=+1} them perfectly preserved was after whole body perfusion of fixative through {/FONT} {BR} {FONT SIZE=+1} a living and pumping heart. {/FONT} {FONT SIZE=+1} {/FONT}
{P} {FONT SIZE=+1} Obviously, it is not likely that pathologists have tissue preserved in this way {/FONT} {BR} {FONT SIZE=+1} from living humans. I guess for argument sake, I would ask the pathologist {/FONT} {BR} {FONT SIZE=+1} weather the swollen mitochondria and other membrane defects could be the {/FONT} {BR} {FONT SIZE=+1} result of the pathology and not poor fixation. {/FONT} {FONT SIZE=+1} {/FONT}
{P} {FONT SIZE=+1} On the other hand, dead cells such as cuticle and cortical cells in hair have {/FONT} {BR} {FONT SIZE=+1} well defined cell membrane complexes that look well preserved even if you {/FONT} {BR} {FONT SIZE=+1} don't fix the hair--although cell organelles like mitochondria are missing. {/FONT} {FONT SIZE=+1} {/FONT}
{P} {FONT SIZE=+1} For perspective, I should tell you that I once worked in the Anatomy {/FONT} {BR} {FONT SIZE=+1} Department at Loma Linda University and the professor there didn't believe {/FONT} {BR} {FONT SIZE=+1} in purified glutaraldehye. He used the 25% stuff and kept it under the lab bench. {/FONT} {BR} {FONT SIZE=+1} His belief was that the older it got the better....something to do with the ratio of {/FONT} {BR} {FONT SIZE=+1} dimer to trimer. I'm not a chemist, so I don't know. I'm sure the Ph.D.'s at the {/FONT} {BR} {FONT SIZE=+1} companies who sell the "good" stuff have their point of view. {/FONT}
{P} {BR} {FONT SIZE=+1} Michael Mead {/FONT}
{P} __________________________________________________________________________________ {BR} Garry Burgess wrote: {BLOCKQUOTE TYPE=CITE} ------------------------------------------------------------------------ {BR} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
{P} First of all, I'd like to thank all of those who cared to respond to my {BR} question, especially Dr. Garber from SPI who gave a detailed and {BR} thoughtful response. And OK, I get the message, it's just not worth the {BR} risk.
{P} But I have another question that perhaps you people also might have some {BR} thoughts on. If I am understanding them properly, some of the {BR} Pathologists here claim that they can tell the difference between a {BR} specimen that was delayed before putting into Glut., resulting in {BR} artifact such as swollen mitochondria, vs. poor fixation as a result of {BR} outdated glutaraldehyde, such as damaged membranes in general. Is this {BR} sort of reasoning valid?
{P} Just curious, {BR} Garry {/BLOCKQUOTE} {/HTML}
Does anybody out there have experience with the {bold} Pixera digital camera system {/bold} . Would appreciate your comments on the image quality, the possibility of converting the images to publication quality prints on a good printer and the level of technical support provided by the company.
My apologies to the list. I sent a private email that seems to have gotten posted to the MDS mail server. (And Eudora is configured not to do that ... the net gremlins at work?)
Phil
****be famous! send in a tech tip or question*** Philip Oshel Technical Editor, Microscopy Today Station A PO Box 5037 Champaign, IL 61825-5037 oshel-at-ux1.cso.uiuc.edu
Would anyone be interested in a used LKB Knifemaker (7800) with cover + six boxes of LKB glass? Fred says it is in excellent condition and that the price would be in the $2500-$3000 range.
If interested, please contact Fred directly at fredl-at-awod.com (Fred G. Lightfoot) or call Fred at (803) 856-8613.
Thanks and regards, Don Cox, Goldmark Biologicals, goldmarker-at-aol.com
Would anyone have an interest in purchasing a used LKB Knifemaker (7800) with six boxes of LKB glass? I think the price will be in the $2500 range. Fred says it is in excellent condition.
If interested, please contact Fred Lightfoot at
fredl-at-awod.com (Fred G. Lightfoot)
or call Fred at (803) 856-8613
Regards, Don Cox Goldmark Biologicals goldmarker-at-aol.com
We section hard tissues containing synthetic biomaterials. While we usually follow a conventional EDTA decal. route, we also sometimes apply a little EDTA to the block face (having embedded in LR White) to decal. "in situ" while sectioning. This alternative seems to work, but as we are using diamond knives it may be purely psychological! I have never seen it written up.
Best wishes, Paul
Dr Paul V. Hatton Lecturer in Biomaterials School of Clinical Dentistry University of Sheffield Claremont Crescent SHEFFIELD S10 2TA
Tel. (0114) 271 7938 Fax. (0114) 2665326 or 2797050
First of all, I'd like to thank all of those who cared to respond to my question, especially Dr. Garber from SPI who gave a detailed and thoughtful response. And OK, I get the message, it's just not worth the risk.
But I have another question that perhaps you people also might have some thoughts on. If I am understanding them properly, some of the Pathologists here claim that they can tell the difference between a specimen that was delayed before putting into Glut., resulting in artifact such as swollen mitochondria, vs. poor fixation as a result of outdated glutaraldehyde, such as damaged membranes in general. Is this sort of reasoning valid?
For an inexpensive target: Obtain a piece of OFHC Cu foil, cut circle slightly larger than target, unscrew the existing target, fold/crimp the foil over the existing target. Even if you have to get the foil from Alfa/Aesar or the like, it should cost only about $100, compared to the several hundred+ vendors want for a "real" target.
Will be curious how well you are able to sputter Cu with the Polaron.
In my previous life doing TEM we would develop and wash negatives for about 5 minutes, then rinse in methanol. Air dry or even a blow dryer on low got us a dry negative in 15-20 minutes. I seem to remember that there was a little clouding of the negative and you can't get too anxious with the hair dryer.
Now, with negative, flatbed scanners one could scan the negative (10 minutes) and print an image on transparency paper with an inkjet (Epson 1440 dpi or a dyesub printer). Haven't tried this with TEM negatives but I know people who do.
Seems like about one hour would do it.
What about a positive TEM film that you could project directly.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Randy,
Sometimes when I want to make a transparency for seminars, or workshops or whatever, I put the EM negative into the enlarger and project it to a small 2X2" size on to yet another piece of EM film that I've cut in half for this purpose. (you need a special enlarger lens to get this small!). Then I cut it to size and mount it in a glass slide mount, and voila, I have supersize black and white slides, with good resolution and contrast. (at least better than 35mm projection slides) But sometimes in the past, if I tried to rush things, I notice that the image turned brown. To fix this problem though, I simply re-fix and re-wash this image, and the brown discoloration (which is probably some residual silver halide and fix) is removed, and all is well again.
Garry
} ---------- } From: rtind-at-siu.edu[SMTP:rtind-at-siu.edu] } Sent: 14 July, 1997 14:51 } To: Garry Burgess } Subject: RE: Rush Lab } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thanks ever so much for all those comments on "burn" marks. As a newbie, I'm grateful for your help. Now I've got a question on sample preparation.
We're currently doing many different types of sample preparation in an "inherited" Edwards Auto 306, which works. But because we do so many different things in it, we're always having to rearrange its "innards" and this is turning out to be a bottleneck. So I was wondering about doing some of the preparations in a separate unit.
In particular, I'm interested in purchasing equipment for plasma etching and carbon coating. (IF--or WHEN-- we can afford it, that is!) So I'm interested in hearing peoples' recommendations, good and bad experiences and so forth. I also have no idea (yet) how much these things cost.
We do the etching to get a better look at those small inorganic particles embedded in a polymer matrix that I mentioned in the EDX spot mode conversation.
(And if vendors would like to contact me, you are welcome to do so! Please do this directly and not via the listserver.)
Title: Optical Microscopy and Imaging in the Biomedical Sciences
When: October 8 - October 16, 1997
Where: Marine Biology Laboratory, Woods Hole, MA, USA
Tuition: $1950 (Includes room and board)
Application Deadline: August 5, 1997
Admission application and information: Carol Harnel, Admissions Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543-1015 (508) 289-7401 Internet: admissions-at-mbl.edu WWW: http://www.mbl.edu
Course Director: Colin S. Izzard, State University of New York -at- Albany Phone: [518] 442 - 4367 EMail: csizzard-at-csc.albany.edu
Course Description:
For Whom: Designed primarily for research scientists, physicians, postdoctoral trainees and advanced graduate students in animal, plant, medical and material sciences. Non-biologists seeking a comprehensive introduction to microscopy and video-imaging will benefit greatly from this course as well. There are no specific prerequisites, but an understanding of the basic principles of optics is desirable. Limited to 24 students.
The eight day course consists of lectures, laboratory demonstrations, exercises and discussions that will enable the participant to obtain and interpret microscope images of high quality, to perform quantitative optical measurements, and to produce photographic and video records for documentation and analysis.
Topics to be covered include: principles of microscope design and image formation bright field, dark field, phase contrast, differential interference contrast, interference reflection, and fluorescence microscopy confocal scanning microscopy and image deconvolution digital image restoration and 3-D reconstruction video imaging, recording, enhancement, and intensification analog and digital image processing and analysis fluorescent probes and ratio-imaging laser tweezers and laser scissors
Applications to live cells will be emphasized; other specimens will be covered as well.
Students will have direct hands-on experience with state-of-the-art microscopes, video cameras, recorders and image processing equipment provided by major optical and electronics companies. Instruction will be provided by experienced staff from universities and industry.
Students are encouraged to bring their own biological (primary cultures, cell lines, etc.) and material specimens and to discuss individual research problems with the faculty.
I've seen someone use this technique to decalcify and section tissue in epon years ago. It worked fine. I also don't know where any written info can be found about it.
By epon, I mean specifically medcast from Ted Pella Inc., I understand this particular media has been discontinued, but I don't think the EDTA decalcification is limited by the media. It only decalcified a few mm-s of surface tissue.
Karen Pawlowski Lab Tech UT Southwestern Med. Ctr. Student/ UT Dallas
On Sun, 20 Jul 1997, P.V.Hatton wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } We section hard tissues containing synthetic biomaterials. While we } usually follow a conventional EDTA decal. route, we also sometimes } apply a little EDTA to the block face (having embedded in LR White) } to decal. "in situ" while sectioning. This alternative seems to } work, but as we are using diamond knives it may be purely } psychological! I have never seen it written up. } } Best wishes, Paul } } } Dr Paul V. Hatton } Lecturer in Biomaterials } School of Clinical Dentistry } University of Sheffield } Claremont Crescent } SHEFFIELD S10 2TA } } Tel. (0114) 271 7938 } Fax. (0114) 2665326 } or 2797050 }
This is prompted by Cynthia Bennett's recent request in regard to plasma etching units. A few years ago, we were briefly interested in such things, particularly in how to remove the surface from a polymeric material without damaging the underlying substructure. However, for our purposes we found that plasma etching, ion mills, etc., would be far too destructive - not only would there be heating problems, but also all those ions running wild would tend to cause a lot of chemical damage.
We had just got round to trying out ATOMIC OXYGEN, generated in a radio frequency discharge, and the first result or two seemed promising, and then with a great bureaucratic reshuffle the owner of the equipmment pulled up sticks and went elsewhere. Does anyone know where such equipment might be obtainable at reasonable cost?
If anyone has experience with this sort of thing, I would be pleased to hear from them.
Thanks in advance,
+------------------------------------------------------------------------+ | 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 | +------------------------------------------------------------------------+
Now that this whole question of expired Glut. and fixation has come up, I'm wondering if perhaps my 2.5% solution that we use here is perhaps suboptimal, and whether or not we might be better off using a strong glutaraldehyde concentration such as 4% for our routine fixation.
Is there anyone else out there fixing human tissue routinely? I would be interested in what concentration of glutaraldehyde that you people are using.
Now that this whole question of expired Glut. and fixation has come up, I'm wondering if perhaps my 2.5% solution that we use here is perhaps suboptimal, and whether or not we might be better off using a strong glutaraldehyde concentration such as 4% for our routine fixation.
Is there anyone else out there fixing human tissue routinely? I would be interested in what concentration of glutaraldehyde that you people are using.
Hello all, I was just asked for answers regarding Canada Balsam, which I hope some member of the group could assist me with, they are..
Questions regarding Canada Balsam, 1) When was it first used for microscopy? 2) How long does it last before degrading? 3) What if any are the aging effects? 4) Does it interfere or affect the sample in any way?
The question was actually asked for a different application other than microscopy, but I thought it would be an interesting discussion none the less.
Thanks
David Dr. David C. Bell Room 13-1018 E-Mail: dcb-at-MIT.EDU Center for Mat. Sci. and Eng. PH: (617) 253-3317 Massachusetts Institute of Technology FAX: (617) 258-6478 77 Massachusetts Ave, Cambridge, MA 02139-4307
We are looking for a used High-Voltage Tank for a JEOL l00. Can anyone help us? Please answer us direct. Thank you. Peter Stolzenberg PESTO INC. pesto-at-erols.com
To all: Can anyone help us getting a used Jeol l00 tank. We would appreciate any leads. Please E-Mail us direct. Thank you! Peter Stolzenberg,Pesto Inc. P.O. Box 648, GWYNEDD VALLEY ,PA 19437 215-699-6160 FAX215-699-5275 E-Mail: pesto-at-erols.com
} Dear Microscopists, } we plann to use TEM Philips 420 for Low-dose work. We will need Low-dose } system for this microscope. Unfortunately Philips said us,that they didn't } produce Low-dose for this kind of microscope,already. } Can you help us please, how or where we could get this Low-dose unit or } how to work in low-dose conditions /to prevent destroing samples during } focusing/ without this system on TEM Philips 420 ? } All your responces or opinions will wery useful for us. } } Thank You very much } Milos } } Milos Motejl } Lab. of Biomembranes } South Bohemian University } Ceske Budejovice } Czech Republic } } motejl-at-paru.cas.cz } tel. 042-038/7775485 } ................................
Philips used to produce free-standing Low Dose modules for their 400 series TEMs - off hand, I don't recall the part number, but if you check around you might find a used one available.
Actually, the easiest approach to low dose is the simplest - work at low magnification. I used to be an applications specialist for Philips, and had the time to experiment with different ways of using a TEM. With a little pre-calibration and experiment, I could repeatedly, with a single micrograph, record the 0.9 nm lattice spacing of crocidolite with the TEM at 10,000 x magnification - the point being that you can record quite high resolution images at low magnification, which gives you sufficient intensity at the film to record the image but considerably reduces the dose to specimen, compared with operating at high mag. Note, that this approach fails if you take the mag into the LM mag range, as the whole optics of the microscope changes at this point, one consequence being a drastic drop in resolution.
You can further reduce the dose to the specimen if you have a STEM unit. Basically the idea is to operate the TEM column in a high mag/high res mode and very low beam intensity but use the STEM unit to image the specimen at low mag/low res, so as to locate a the region of interest.
With a standard STEM set up, you can only image a small part of the field of view. However, if you have a computer connection to allow you to control beam position, it is straightforward to produce a short routine that will step the illuminated area across the whole area covered by the film. With this approach, I have 'easily' recorded diffraction patterns and images from parafin wax.
With the same sort of computer control set up, it is not much more difficult to duplicate most of the functions of the low dose unit, except that you can't change the illumination focus - but that can be done manually at the appropriate step in the sequence.
Garry Burgess wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Now that this whole question of expired Glut. and fixation has come up, } I'm wondering if perhaps my 2.5% solution that we use here is perhaps } suboptimal, and whether or not we might be better off using a strong } glutaraldehyde concentration such as 4% for our routine fixation. } } Is there anyone else out there fixing human tissue routinely? I would } be interested in what concentration of glutaraldehyde that you people } are using. } } Garry
Here in the EM Facility at Boston Medical Center, we use 2.5% glut in 0.2M cacodylate buffer for all human specimens; the fix lasts for several months in the fridge and yields very high quality results if the tissue is fixed promptly at the biopsy site. If a surgical specimen has been held overnight in the cold and is then sampled for EM in the morning, the preservation suffers but is still suitable for diagnosis.
Dr. Tom Christensen Director, EM Facility Boston Medical Center Boston, Mass
I do not know if you have gotten any other replies on this. We do failure analysis on integrated circuits. There are two types of "short circuits". One is a resistive short, and the other is a leakage path where hole-electron recombination occurs. Resistive shorts can be located with liquid crystal methods that sense the thermal dissipation. (There are other, more complex, methods, but we use liquid crystal extensively) Leakage paths can be located with systems based on "night vision" technology that was developed for the military. Hole-electron recom- bination releases the excess energy as photons, and the light amplification allows the imaging of the emission site.
Darrell Miles IBM Microelectronics Test and Analytical Services http://www.chips.ibm.com/services/asg
A tentative okay has been given for a session on corrosion casting for the 1998 meeting in Atlanta. We would urge any of our Mercox users who are interested in presenting a paper to contact us at Ladd Research or Dr. Fred Hossler at:
Dr. Fred Hossler Professor of Anatomy East Tennesse State University Johnson City, TN 37614 e-mail SEMTEMman-at-aol.com
Methadology is of particular interest.
We would also like to have some opinions on the advantages/disadvantages of using clear, blue or red mercox for casting.
In March 1997 we requested protocols and references for staining semi-thin sections of vertebrate tissues embedded in plastic; our original standard stain was toluidine blue.
We received 14 responses to this request, which we have edited to a ~7 page file. Responses included stains such as hematoxalin, alcian blue, eosin, Mayer's mucicarmine, methyl green, methylene blue/azure II, basic fuschin, and Stevenel's blue. A copy of the file is available upon request by e-mail. We have begun using a polychrome stain (see Van Reempts and Borgers, 1975, Stain Tech. 50:19-23) with pleasing results.
Christine Roy and David Hall Albert Einstein College of Medicine Bronx, NY 10461
Dr. Steven Barlow EM Facility/Biology Department 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/EM_Facility
I am interested in finding out what people are using for 4"x5" SEM film. I have used Polaroid Type 55 P/N and Kodak type 4427 Commercial Film. At $1.50 to $2.00 per exposure, both are a little too expensive for student use (i.e. low good photo to bad photo ratio). Can anyone recommend a less expensive alternative? My predecessor used to buy 200 foot rolls of surplus aerial photography film (4 or 5 inches wide) which he would cut to the proper length. Final cost was less than $0.05 per sheet. Any similar suggestions?
Bob
Dr. Robert R. Wise Department of Biology University of Wisconsin-Oshkosh Oshkosh, WI 54901
(414) 424-3404 tel (414) 424-1101 fax wise-at-uwosh.edu
Nora Pratta Centro Regional de Investigacion y Desarrollo Santa Fe - Argentina Inquired about doing SEM/BSE/EDX on particulates in oil and oil/water mixture.
If the oil or oil water mixture can be 'frozen' at liquid nitrogen temperatures by rapid freezing in a metal block or propane jet freezer th= en transferred to a cryo ultramicrotome where you could prepare frozen sections for examination on an SEM cold stage. =
This is a very expensive but technically superb method. An alternate woul= d be to freeze dry the particulates out of the section onto a carbon substrate or other suitable substrate. You will lose some of the distribution information in the X-Y plane but should resolve the Z distribution represented by your sections themselves.
There are some commercial labs with this equipment and some private companies that may collaborate if the subject interests them.
You may contact me off line or visit the Web Site: =
http://www.RMC-Scientific.com/microtomes/ =
We are a commercial manufacturer of all of the instruments listed above.=
Steve Miller Director of Sales RMC 3450 S. Broadmont, Suite 100 Tucson, AZ 85713 Tel 520-903-9366 Fax 520-903-0132
"Inter/Micro" is a meeting, now in its 49th year, held annually in Chicago and hosted by the McCrone Research Institute (McRI). I am attending the meeting as I have almost every year for the past 25 or so. However it is a meeting little known to those without regular contact with McRI. As a way of introducing this under-attended meeting to a wider audience I thought the readers of this list might find it informative to have a summary of the technical highlights and other goings-on at Inter/Micro. If I can, I'll write a daily summary every evening and post it to this list. Otherwise I'll just summarize and post as time permits. I'll mention two or three of those papers I found most interesting each day.
This year approximately 60 technical presentations are scheduled. A single meeting room is used and there are no parallel sessions. One has an opportunity to hear all of the papers and I find it quite valuable to sit in on papers which might not be in an area of my own personal focus. This "cross-fertilization" of ideas and techniques is often the most enlightening thing that I experience at technical meetings and I am glad they've continued the tradition at Inter/Micro. There will also be an exhibition of products and equipment, as is usual at such meetings. A full program is available at McCrone's web page, http://www.mcri.org. I won't duplicate that here but I'll just mention that the themes for the various sessions for the week are as follows: Monday: General Microscopy Tuesday am: Instrumentation Tuesday pm: Techniques Wednesday: History and Art Wednesday Eve: Dinner in Conjunction with the State Microscopical Society of Illinois (SMSI) Thursday: Forensic Microscopy Friday: A Tutorial on Dispersion Staining
The highlights (for me) of Monday's program included the following.
The program was opened with a talk by Brian Ford on "Crytosporidium, a New Threat from Water Supplies." Brian discussed how Cryptosporidium is representative of a "new" class of microorganisms threatening our modern society. Of course, it is not new at all however a combination of factors is working together to make many existing organisms newly hazardous. These factors include new practices and factors coming with technological advances such as the wearing of contact lenses which provide a previously non-existing environment in which certain organisms can thrive. Another factor is the emerging resistance of some organisms to existing treatments. Previously "eradicated" threats are reemerging as "new" threats again. In the case of Cryptosporidium, we have an organism causing severe but usually non-fatal intestinal distress that we are immune from after the initial exposure and bout of sickness. But it has become so "rare" as a contaminant that few of us received the immunizing infection early in our lives, leaving large segments of the population subject to infection when water treatments fail or other sources of the protozoa present to the population.
Another highlight today was a paper by John Wuepper of Whirlpool entitled "Problem Solving via Analytical Microscopy: What is it Really Worth?" At Inter/Micro we have on many occations over the years lamented the under-appreciated and under-valued status of the work we do on behalf of industry and society at large. John suggested an excellent technique that we can use if we revise our thinking and the presentation of our work to the corporations or agencies we serve. He demonstrated with several examples that the "leverage" obtained from an investment in microscopical problem solving is often enormous, in the range of 25 to 5,000 in the examples he gave. By "leverage" he meant the ratio of the cost of the process to the savings enjoyed by the company as a result of the work done. The "investments" may range from a few hundred dollars to a hundred thousand or more, but the return and leverage is enormous. In one instance he cited microscopy saved a company more than $500,000,000 dollars. (Those figures will catch the eye of even the most myopic bean-counter - my editorial, not John's!) A lively discussion followed John's presentation and it is clear that his approach of communicating with our host-employers in terms they understand and respond to is critical to the success and growth, sometimes just the survival, of a microscopy laboratory.
Jan Hinsch or Leica gave one of the most beautifully illustrated as well as educational talks of the day when he spoke on "What Pleurosigma can Tell the Microscopist." Pleurosigma Angulatum, a species of diatom, has long been used as a microscope test object for evaluating the quality of higher numerical aperture objectives. Jan's talk was one of those wonderful half-hours where an audience gets to enjoy not only an aestheticly beautiful talk but one that, through the instructional insight of the author, makes crystal clear some technically difficult fundamental principles. In the case of today's talk, those principles dealt with the theoretical resolution limits of the light microscope and I, along with many others in the audience I think, came away with with a vastly better understanding of what had heretofor been a baffling subject.
Another fascinating talk today illustrated the diversity of topics we regularly enjoy at Inter/Micro. Ryan D. Tweney of the Department of Psychology at Bowling Green State University spoke on "Cognition and the Microscope." Ryan discussed and illustrated many of those murky processes that stand between knowledge or observation on the one hand and understanding on the other. It was apparent that he was only able to scratch the surface of a subject which we would all benefit from knowing much more about and I, for one, hope we'll see him back regularly in the future.
I'll try to come back tomorrow with another update! Right now I've got friendships to rekindle and think I hear the calling of a lonely brew with my name on it.
Thank you to those of you who replied regarding LM mounting medium. It appears as if oxidants in the medium are the main culprits. I am listing these replies below.
One point worth mentioning about the common use of epoxy resins as a coverslip mountant is that its refractive index is not 1.5 (I'm not sure of its exact value). One may be able to get away with it working at low numerical apertures, but it will take its toll at 0.65 and above.
Thanks again!
James Wesley-Smith EM Unit University of Natal Durban, South Africa
I learned a trick from a DuPont-Sorvall rep (that tells you how long ago that was!) that retards or eliminates ALL fading caused by oxidants in the mounting medium. Add 1-2% BHT (the preservative used in bologna, hot dogs, etc.) to any mounting medium. -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- I have been using ENTELLAN which has ND20 1.49-1.5 and gives excellent preservation of toluidine blue stained plant tissue for at least 4 years. I get it from Electron Microscopy Sciences who have a good list of mounting media with refractive indicies in their catalogue. -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- Hello fading is the problem. We use these toluidine blue sections for histology courses and we haven't find yet the no-fade mounting medium. Our best choice is DEPEX, manufactured by GURR. Avoid EUKITT, fading occurrs in a matter of hours. Some collegues have used cured epon, but it is a time consuming process and fading does occur. -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- I think the fading is caused by oxygen (which very easily diffuses through hydrophobic media like resins). We routinely leave such preparations uncovered, and add a drop of immersion oil and coverslip to photograph. This prep is easily soaked off in xylene to restain if necessary. -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- I don't know about the refractive index, but I've used Epoxy resin as a mountant quite successfully. It doesn't seem to fade Toluidine Blue semi-thins.
DePeX is not too bad but I have had some fading over long periods. -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
I have been working with plant material embedded in Spurr resin for many, many years. One micron sections were stained with toulidin blue or other specific stains and permanent mounted with Permount, Fisher Scientific, and no fading for decades. I have not seen any fading using DePeX, but Spurr resin sections usually get very wrinkeled.
-at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- We use the following protocol for toluidine blue (TB) staining and permanent mounting: Frozen or dewaxed abd hydrated paraffin sections 0.1% TB in acetate or phosphate buffer (generally at pH 2-3 for sulfated glycosaminoglycans) 5 min Rinsing in buffer Precipitation with a 6:1 mixture of 2% aqueous KI and Kferricyanide 2-3 min Mount with a drop of 25% aqueous gum arabic containing 2% fructose without coverslip! Form a layer of te mounting medium using a glass rod. Let the gum arabic layer dry at room temperature in horizontal position (it takes generally one night). The refractive index of the dried gum arabic is practically identical to that of the glass. Mount in DPX or Canada using a coverslip. This procedure is good to produce permanent metachromatic staining. Dimethylmethylene blue (DMMB) is a better metachromatic dye (Aldrich Co, or SERVA) The protocol is similar, except the poststaining stabilization. For this purpose, 2% aqueous ammoniummolybdenate is used. DMMB is a very strong metachromatic staining. It is very useful for mast cells, cartilage, sulfomucins. In many cases, we use it successfully in 0.05 or 0.01% aqueous solution for 5-10 min.
For more inforations, see Modis, L.: Organization os the Extracellular Matrix: A Polarization Microscopic Approach. CRC Press, Boca Raton, 1991. Chapter 12.
Kindly forgive me if these references have already been posted, but the answers to many of these processing protocol queries can also be found in a series of articles by Coetzee and van der Merwe, viz.
J Coetzee and CFvan der Merwe (1984) Extraction of substances during glutaraldehyde fixation of plant cells. Journal of Microscopy 135, Pt2, pp 147-158.
J Coetzee and CFvan der Merwe (1985) Penetration rate of glutaraldehyde in various buffers into plant tissue and gelatin gels. Journal of Microscopy 137, Pt2, pp 129-136.
J Coetzee and CFvan der Merwe (1986) The influence of processing protocol on the ultrastructure of bean leaf cells. South African Journal of Botany, 52, pp 95-99.
These articles are compulsory reading for our trainee microscopists, since they dispell many processing 'myths'.
James Wesley-Smith EM Unit University of Natal Durban, South Africa
We want to analyse the intensity of ED-patterns of organic specimen. For this we used ELD, a program which is included in the CRISP packages from Calidris. Now we want to compare the intensity data from ELD with the intensity getting from other programms. Does anybody knows some programs, which we can used for intensity estimation? Can you tell something about prices and the possibility to get such programms!
Hans Kothe Working group Dr. Voigt-Martin Universit=E4t Mainz =20
25th Scottish Microscopy Group Symposium (First Circular)
Stakis Dunblane Hotel, Dunblane. Wednesday 12 November 1997.
This the SILVER Scottish Microscopy Symposium will take place at the=20 above venue and the Organising Committee have arranged a Scientific=20 Programme which we hope will appeal to as many microscopists as=20 possible. Also a celebratory meal will mark this anniversary.
The following topics have been selected:
Environmental Scanning Electron Microscopy - Dirk van der Vall,=20 Eindhoven, Holland.
Advances in Confocal Microscopy - Tony Wilson, Oxford, England
Stereology - Vyvyan Howard, Liverpool, England
Cryo/Immunocytochemistry - Jeremy Skepper, Cambridge, England
These invited talks will be interspersed with short presentations. We=20 would welcome offers of short (10-15 minute) talks which deal with any=20 aspect of microscopy and in particular electron microscopy. There is an=20 abundance of useful techniques and protocols in daily use; if you think=20 that you have something which others could adopt or benefit from,=20 please send us your name and a brief title for your presentation to Ian=20 Roberts at {irober-at-scri.sari.ac.uk}
These meetings are enjoyable, interesting and useful and an=20 opportunity to meet and share ideas with fellow microscopists. The cost=20 will be =A320 (pounds).
Celebratory Dinner (evening)
To mark the 25th Anniversary of these meetings, we hope to arrange an=20 evening dinner to which all delegates and partners are invited to attend.= =20 The separate cost of this will be =A325.00/head, and a favourable rate of= =20 =A3100 per night/ per couple for dinner, bed and breakfast at the hotel has= =20 been negotiated. If you wish to attend this evening function, please=20 contact Martin Maxwell at {MARTIN.MAXWELL-at-BBSRC.AC.UK}
A second circular will be sent in the future given details of all talks. Al= so=20 there is a web page at http://www.abdn.ac.uk/~nhi691/smg97.htm that=20 gives more information.
Kevin Mackenzie Tillydrone E.M. Unit University of Aberdeen Tillydrone Avenue Aberdeen AB9 2NT
Tel 01224-272847 Fax 01224-272396 Web site- http://www.abdn.ac.uk/~nhi691/
---------------------- Kevin Mackenzie k.s.mackenzie-at-abdn.ac.uk
We used to commonly use a Kodak film called Ektapan in 4x5 size. It is developed in D-76. usually, although other standard film developers could also be used. The advantage is that it's much cheaper than Polaroid PN 55, at about $20 per 25-sheet box (last time we checked the price). The disadvantages, of course, are that a developing set-up and darkroom are required and you don't get an automatic study print.
Actually, it would seem that any common 4x5-inch film could be adapted to SEM use, depending upon contrast requirements. T-Max 100 or 400, Plus-X Pan, Ilford, or Agfa films, etc., all should work. All of these films should be readily available and all use a variety of common developers.
Hope this helps.
Randy Tindall Center for Electron Microscopy Southern Illinois University at Carbondale
Since my initial request for the x-ray wavelengths file I have since found a Microsoft Access database file which was originally provided by John Donovan (UC Berkeley). I have also received request for posting the file for FTP (... altho one reply indicated the original "Fiori" file was available at ftp://www.anc.anl.gov ...), and the John has since indicated his database which also includes higher order lines is generally available. I can make John's MDB file and my Excel (XLS v2.1) file available via "anonymous" FTP. My XLS file is only slightly different, i.e., it has been sorted by "Z" and "N" ... Lastly, a word about the FTP site. It is actually a magneto-optical drive and used for archiving image files. As a MO drive it will offer these specific files only until this cartridge fills and I have to put in another (approximately 1 week). I can make these files available in the future upon request.
You can point your browser at ftp://whitewater.uoregon.edu/share/cameca/
or FTP anonymous to whitewater.uoregon.edu
and look into the "/share/cameca/" directory
and find John's original Access file (xray.mdb, 704kb) ... you can also find the Excel file I created with it, but it is 2Mb. I couldn't get Access to save the file as XLS ... it created the file but the cells were empty (?) ... I copied and pasted instead ... you may not be able to C&P if you don't have enuf memory. Let me know if you have any problems ...
TIA & cheerios, shAf {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/epmahome/
Long storage in glut is not good - the membranes are still permeable since glut does not fix lipid. There will be an exodous of material and changes. But - prefix, wash, postfix with osmium stabilizing the membranes and the lipids, and store in buffer (not in alcohol - alcohol removes osmium) for as long as needed. There is some movement of osmium, but unless the storage is in a solvent, it is neglible. Bye, Hildy
I need some help identifying clays using SEM. Do you know of references, textbooks, etc. with pictures that would me identify common clays i.e. illite, kaolinite, smectite, chlorite, etc.
The knifemaker available to me is a LKB Type 7801A from LKB Instruments Inc. out of Rockville Maryland, but the company has since been taken over by Leica/Leo. The knifemaker has two wheel type gauges that adjust the tension from opposing sides on the glass piece. I can make glass squares, but when I try to cut the square into the two knives, I don't get the correct shapes, even though I have tried systematically changing the settings on the two gauges many different ways. Typically, the "sharp" edge may be chipped, the reflection line in the glass is going in the opposite direction to what it should and the opposite edge to the sharp edge may be either sharp as well or too thick a blunt edge. That's just one of the two knives; its' counterpart often has two "sharp" edges. Can anyone with any direct experience with this model of knifemaker provide me with any advice on how to adjust the settings to produce two glass knives?
Thanks in advance for any assistance that can be provided.
The knifemaker available to me is a LKB Type 7801A from LKB Instruments Inc. out of Rockville Maryland, but the company has since been taken over by Leica/Leo. The knifemaker has two wheel type gauges that adjust the tension from opposing sides on the glass piece. I can make glass squares, but when I try to cut the square into the two knives, I don't get the correct shapes, even though I have tried systematically changing the settings on the two gauges many different ways. Typically, the "sharp" edge may be chipped, the reflection line in the glass is going in the opposite direction to what it should and the opposite edge to the sharp edge may be either sharp as well or too thick a blunt edge. That's just one of the two knives; its' counterpart often has two "sharp" edges. Can anyone with any direct experience with this model of knifemaker provide me with any advice on how to adjust the settings to produce two glass knives?
Thanks in advance for any assistance that can be provided.
People, Does anyone know where to get uranyl formate? My previous suppliers no longer make it-why? I do have the acetate, but the formate works better with microfilaments. Any leads are welcome.
Last winter when I was making some changes to our tissue processing/embedding protocol (which has been around here forever it seems), I began asking questions about EDTA decalcification. One of our professors told me that the EDTA decalcification was found to produce the fewest if any artefacts IF (1) you use the disodium salt (not the tetrasodium), (2) it is done at 4 C (not room temp), AND (3) decalcification is complete in 7-8 days. He claims that tissue pieces which take longer than that begin to loose their membrane integrity. I have no references for this but he has always been a reliable source of information in the past.
Pat Hales McGill University Dept. of Anatomy & Cell Biology hales-at-hippo.medcor.mcgill.ca
If some of you EDS practitioners (yes, I'm a WDS practitioner) could help me with this I would be most appreciative:
1. presently I am involved in a project in which I have to examine zinc arsenides - the process by which these are made can, at times, also produce elemental Zn, ZnO and elemental As (yuk!, and then some)
2. I mainly characterize these beasties by x-ray diffraction, but in this particular inspection I am doing some SEM work too
3. the samples I am looking at are suspected to be mainly the zinc arsenide phase(s), however, I get EDS spectra with highly variable Zn/As - in fact, more wide-ranging than expected for various of the possible zinc arsenides
4. I suspect that the Zn/As variation has as much to do with loss (diminishment) of the As signal (due to absorption?) as it does with variation in the ZnAs
5. to that end (#4), I performed a test yesterday in which I examined just one crystal type (based on morphology) in areas of the sample where these crystals were at the "surface" of the sample and at various "depths" within holes and/or depressions.....basically, the deeper in the "hole" the lower the As signal
6. concurrent with the dimished As, I also observed diminishment of Zn L-alpha line (also a low-energy x-ray) such that Zn-K/Zn-L was also highly variable - would you consider the latter a further indication of absorption?
In the interim, as I await your response, I intend to run x-ray diffraction on those samples with "apparently" low As (some are even As "absent") to "confirm" whether or not the Zn or ZnO phases could be present. Together (my XRD and preliminary SEM, plus your sage advice) I hope we can resolve this.
Thanks, in advance!!
Winton
P.S. if this message "surfaces" twice, please forgive me...the first time I sent it I directed it to the listserver.....once discovered, some kind soul might take it on himself/herself to forward it back to this list
Dr. Winton Cornell Senior Research Associate & Supervisor, Microanalysis Laboratory Department of Geosciences The University of Tulsa 600 South College Tulsa, OK 74104-3189
Hi All: I recently put a message on the listserver requesting information on a LogEtronics enlarger. Some responses I received suggested that I should think about using a digital camera instead to capture images from negatives and then process the images and then send them to a printer. I would like some feedback from users. Is the quality close to that of film? Does anyone have a system that they are extremely happy with? Are there any commercial systems available? Please let me hear your experiences.
Thanks,
Michael Coviello EM Lab Manager The University of Texas -at- Arlington Arlington, TX E-mail coviello-at-mae.uta.edu 817-272-5496
Since my last post I have been working with the XLS spreadsheet file and have added absorption edges, font highlighting, and Chuck's and John's references. The new file name is xray_MS.XLS and is zipped as xray_MS.ZIP ... Like I said before this will be a temporary FTP location for this file ... if Nestor is watching this thread, maybe he'll put the zipped file on the MAS FTP site. For those interested, I'm in the process of adding Cameca spectrometer sine-theta values and creating a PDF file for the purpose of having this data immediately available for on-line browsing ...
You can point your browser at ftp://whitewater.uoregon.edu/share/cameca/
or FTP anonymous to whitewater.uoregon.edu
and look into the "/share/cameca/" directory
Let me know if you have any problems ... and PLEASE let me know if you find any errors ...
TIA & cheerios, shAf {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/epmahome/
Sharon, I would replace the cutting wheel first and then see what happens. The systematic approach should work.
cheers
Edward J. Basgall, PhD The Pennsylvania State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618
{The knifemaker available to me is a LKB Type 7801A from LKB Instruments Inc. {out of Rockville Maryland, but the company has since been taken over by {Leica/Leo. The knifemaker has two wheel type gauges that adjust the tension {from opposing sides on the glass piece. I can make glass squares, but when {I try to cut the square into the two knives, I don't get the correct {shapes, even {though I have tried systematically changing the settings on the two gauges {many different ways. Typically, the "sharp" edge may be chipped, the {reflection line in the glass is going in the opposite direction to what it {should {and the opposite edge to the sharp edge may be either sharp as well or too {thick a blunt edge. That's just one of the two knives; its' counterpart {often has {two "sharp" edges. Can anyone with any direct experience with this model of {knifemaker provide me with any advice on how to adjust the settings to produce {two glass knives?
{Thanks in advance for any assistance that can be provided.
You will get lots of opinions on this. Here's mine. After spending countless hours in the darkroom over the years, I've converted enthusiastically to digital imaging, and now would prepare any serious final labeled images with Photoshop, making the final prints on a photographic quality printer. Does that mean that all original EM pictures must be taken digitally? I don't think so. If you think about it, only a small percentage of the EM images you take are apt to end up in publications, projection slides or other serious uses. I find it much easier to store numerous EM negatives as 3-1/4x4" sheets of film in glassine envelopes than to fiddle with the multiple zip disks that are necessary to store all the EM digital images in files large enough to allow the resolution that may be necessary later on (the resolution level that we take for granted in film negatives). So I would take them initially on film (even though our department has a Philips EM100 fitted with a CCD camera for digital imaging), and would observe the negatives on a viewer (or on study prints) to decide what will be used for the final product (publication figures, projection slides, etc.). For the final pictures, I would generate digital images by scanning the chosen negatives (with Leafscan 45 scanner) or scanning carefully-prepared prints (with a Hewlett-Packard ScanJet II CX scanner), then crop, arrange, label and otherwise fine tune the pictures with Photoshop 4 on a Power Mac 7600. For photographic quality printing, we use a Kodak XLS 8600 PS printer.
For example, a very complex figure was prepared from multiple darkroom prints, from which numerous small rectangles were cut out and mounted, each showing polysomes at 100 kX mag. To convert to digital image, I scanned the complex figure with the HP ScanJet, enlarging (exaggerating) the image size so the file was 10-15 MB (to provide good resolution). The scan was done without scanner sharpening (sharpening would be done in Photoshop). The file was saved as a TIFF file, and taken up in Photoshop 4. Crop properly (rotate slightly, if necessary). The brightness/contrast of the rectangles varied somewhat, so each was selected (marquee), and Image} Adjust} AutoLevel was used to normalize (be sure white=~5-7% and black=95% in Image} Adjust} Levels [or Curves] eyedroppers). Sharpen whole figure with Filter} Sharpen} Unsharp Mask with amount ~150-200%, radius 1, threshold 0. Dust or other small blemishes (that are clearly of no scientific interest) can be removed with Filter} Noise} Dust & Scratches (after minimal selection with marquee), using radius 1-5 (no more than necessary to remove). Size (Image} Image Size) the figure to width (or height) required by the journal, and set resolution to 400-600 ppi. Put in figure number, size bar, and labels, using layers (not directly on EM image). Print on Kodak XLS printer. The image quality and detail compares well with work done in the darkroom. More and more journals now allow you to submit such digital image files for the final reproductions after a paper has been accepted.
Kent (A. Kent Christensen, University of Michigan, akc-at-umich.edu)
-------------------------------------------------
On Tue, 22 Jul 1997, Mike Coviello wrote:
} I recently put a message on the listserver requesting information on a } LogEtronics enlarger. Some responses I received suggested that I should } think about using a digital camera instead to capture images from negatives } and then process the images and then send them to a printer. I would like } some feedback from users. Is the quality close to that of film? Does anyone } have a system that they are extremely happy with? Are there any commercial } systems available? Please let me hear your experiences. } } Michael Coviello } EM Lab Manager } The University of Texas -at- Arlington } Arlington, TX } E-mail coviello-at-mae.uta.edu } 817-272-5496
The Microstructure and Microanalysis Program in the Characterization and Environmental Laboratory at GE Corporate Research and Development, Schenectady, NY, has an opening for a Polymer Microscopist at the Lead Professional level. The primary duties associated with this position involve the execution of research projects using Transmission Electron Microscopy (TEM) to determine the structure of polymer blends and coatings. Additional duties may involve research conducted using TEM to study other materials, including metals, ceramics, or composites, or using other microscopy techniques, including Acoustic Microscopy and Atomic Force Microscopy.
The Characterization and Environmental Technology Laboratory is involved in research into the structure and composition of materials in support of development programs both at GE CRD and at GE businesses. Staff members are expected to work independently with a high level of expertise, and to become involved with a number of major project teams. Good communication skills, both written and oral, are extremely important.
The minimum requirements for this position are a MS in Materials Science or a closely-related field and some prior experience with Transmission Electron Microscopy. Demonstrated experience in polymer materials science and characterization is also highly desirable.
Resumes and other information can be sent to:
Ernest L. Hall Manager, Microstructure and Microanalysis Program Room K1-2C12 GE Corporate Research and Development PO Box 8 Schenectady, NY 12301 Fax: 518-387-6972 E-mail: hallel-at-crd.ge.com
I'm going to a new position (PPG Industries) and I'm planning to implement a digital darkroom instead of a conventional darkroom. The advantages are tremendous.
At the Materials Directorate at Wright Lab, WPAFB, we have been using a Leaf 45 negative scanner. They are hard to find, but there is a company that has updated it and is now selling it:
I sent out a message several weeks ago on a web site that has information on a collection of negative scanners, flatbed scanners, and some drum scanners:
http://www.foto.unibas.ch/scanners.html
Scitex used to make the Leaf, but now they have their flatbed smartscanners:
http://www.scitex.com/
Here's a site for information on doing it digitally:
A site for drum scanners: http://www.budde.com/products.htm http://www.budde.com/print/magic.htm http://www.budde.com/print/scanview.htm http://www.budde.com/print/sm11000.htm
Our Leaf system is hooked into a MAC system with a lot of RAM, disk space, MO drive, ZIP drive, and access to our LAN. This gives a lot of flexibilty with different users. We bring in the images with a 16 bit format into Photoshop, adjust the levels, convert the image to 8 bit grayscale with appropriate gamma processing, invert the image into a positive print and save. If the intermediate 4 x 5 setting of the Leaf 45 is used, a TEM negative can be digitized with about 1200 dpi. If the image is printed to a 300 dpi grayscale printer (i.e. sub-dye), then that gives an enlargement of 4x at the printer. The Leaf system is capable of much higher pixel resolution and can be used to digitize a very small area on the negative which can be printed with the printer's 300 dpi setting providing a very high enlargement factor. Photoshop has all the tools that could be done in the darkroom but are rather tedious to do such as unsharp masking. Fixing scratches on negatives (mine never have them), dodging, burning an other things that are required for printing a good TEM negative can all be done in a few minutes. In addition, it can be used on both MAC and PC platforms. John Russ's Image Processing Toolkit provides plugins for Photoshop that can provide image processing and stereology: http://members.aol.com/imagproctk/index.htm
I put the scale markers into the print in layers, thus preserving the original scanned image and saving in a TIF format provides the image in a compatible format for other programs. I particularly like printing from Powerpoint onto our Kodak 8650 printer. With Powerpoint, I copy several images of the Tif file on one page (or several differnt files) and print. Output quality if very good.
You need a fast computer with large RAM and large hard drives, a good size monitor (minimum 17"), the scanner, pub-quality printer, and a portable-disk large format disk drive. A 600 dpi laser printer is quite useful also. I also use a flatbed scanner to digitize 6 TEM negatives in a Neg-a-file sheet and digitize the page at 150 dpi. I also digitize my datasheet notes in lineart format. These images are then put into my ThumbsPlus Image database (shareware Program ~$50 for PC and Macs) I then have instant access to my images. It sure beats making proof sheets and processing them.
All of this would probably be cheaper than going with a LogEtronics.
-Hopes this helps.
-Scott
} ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com -----------------------------------------------------------------------. } } Hi All: } I recently put a message on the listserver requesting information on a LogEtronics enlarger. Some responses I received suggested that I should think about using a digital camera instead to capture images from negatives and then process the images and then send them to a printer. I would like some feedback from users. Is the quality close to that of film? Does anyone have a system that they are extremely happy with? Are there any commercial systems available? Please let me hear your experiences. } } Thanks, } } Michael Coviello EM Lab Manager The University of Texas -at- Arlington Arlington, TX E-mail coviello-at-mae.uta.edu 817-272-5496 }
I would like to summarize some of the answers I received concerning the labelling of actin at EM level in non-muscle cells, namely chondrocytes in the rabbit growth plate.
Rosemary White labelled actin in plants using monoclonal C4 anti-actin from ICN on LR White sections (Protoplasma 131:153-165 and 150:72-74). This antibody is mouse IgG against human actin and recognizes a common actin epitope in many species. Kirk Czymmek was successful using the antibody N.350 from Amersham to label actin at the EM level in fungi (J Microscopy Vol 181, Feb 1996 pp 153-161; Protoplasma 163, pp. 199-202). This is a mouse IgM against chicken gizzard actin and has been shown to label human, monkey, chicken, rat, etc actin, therefore binding to a highly conserved region of actin. BTW, we ordered this antibody today and hope to have some results shortly!
Since we got nice labelling with phalloidin-BODIPY FL in the confocal, we wanted to use rabbit anti-BODIPY FL antibody (Molecular Probes) followed by anti-rabbit-gold to reveal the actin at EM level. This was not successful. Molecular Probes could not give us any references of anybody using this antibody for immunohistochemistry. Tamara Howard commented, that though she has not used the anti-BODIPY, she had a dismal luck with antibodies to FITC and Texas Red from various sources at the EM level.
Thanks to everybody who took the time and responded to my questions. This list is such a wonderful way of sharing experience! I would never think of looking in Protoplasma, since Medline does not bring it up!
Sarka Lhotak EM Facility, McMaster University Hamilton, Ontario, Canada
Dear Winton, If you look at particles in holes or depressions, the edges of the hole will preferentially absorb the lower energy x-rays while the higher energy ones can penetrate the edges of the hole. If you look at the As Ka line with the SEM at 25 or 30 kV it will be less affected than the As L line. You will also get better quantification. As with WDS, a flat, smooth sample would also help. You wrote: } Micro-colleagues: } } If some of you EDS practitioners (yes, I'm a WDS practitioner) could help me } with this I would be most appreciative: } } 1. presently I am involved in a project in which I have to examine zinc } arsenides - the process by which these are made can, at times, also produce } elemental Zn, ZnO and elemental As (yuk!, and then some) } } 2. I mainly characterize these beasties by x-ray diffraction, but in this } particular inspection I am doing some SEM work too } } 3. the samples I am looking at are suspected to be mainly the zinc arsenide } phase(s), however, I get EDS spectra with highly variable Zn/As - in fact, } more wide-ranging than expected for various of the possible zinc arsenides } } 4. I suspect that the Zn/As variation has as much to do with loss } (diminishment) of the As signal (due to absorption?) as it does with } variation in the ZnAs } } 5. to that end (#4), I performed a test yesterday in which I examined just } one crystal type (based on morphology) in areas of the sample where these } crystals were at the "surface" of the sample and at various "depths" within } holes and/or depressions.....basically, the deeper in the "hole" the lower } the As signal } } 6. concurrent with the dimished As, I also observed diminishment of Zn } L-alpha line (also a low-energy x-ray) such that Zn-K/Zn-L was also highly } variable - would you consider the latter a further indication of } absorption? } } In the interim, as I await your response, I intend to run x-ray diffraction } on those samples with "apparently" low As (some are even As "absent") to } "confirm" whether or not the Zn or ZnO phases could be present. Together } (my XRD and preliminary SEM, plus your sage advice) I hope we can resolve this. } } Thanks, in advance!! } } Winton } } } P.S. if this message "surfaces" twice, please forgive me...the first time I } sent it I directed it to the listserver.....once discovered, some kind soul } might take it on himself/herself to forward it back to this list } } } Dr. Winton Cornell } Senior Research Associate & Supervisor, Microanalysis Laboratory } Department of Geosciences } The University of Tulsa } 600 South College } Tulsa, OK 74104-3189 } } phone: 918-631-3248 } fax: 918-631-2091 } e-mail: wcornell-at-centum.utulsa.edu } } } } Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
Continuing my reports on Inter/Micro 97: Today's sessions focused on Instrumentation (AM) and Techniques (PM).
In the morning session, James M. Landrigan III of Polaroid discussed their new Digital Microscope Camera ("DMC"). I have been anxious to see what Polaroid has been up to and I am not surprised to report that their new system will mark a significant addition to the development of digital imaging. I won't risk mis-quoting the specs here since I didn't yet get their literature, but I'll just mention that the system includes a dedicated digital camera with a standard C-mount lens attachment which should make it readily attachable to a range of microscopes (and other equipment). The system also incorporates a larger (12.5mm?) pixel array than competing systems which they claim results in an improved signal-to-noise ratio. The camera includes more than 1,000,000 pixels and produces images of either 800X600 or 1600X1200 pixels (other formats too?) Part of the available resolution is from software interpolation, which I'm not wild about, but I couldn't get clear just what the actual pixel array was so you'll want to get more details from them. Also, one factor which concerns me at first hearing is that they use a rectangular pixel, not a square one. I'll want to ask them why that choice was made. The price is about $6,000 for the camera and software system which, while not cheap, certainly seems reasonable. All-in-all, I'm happy to see Polaroid step into this field. Competition among the giants can only help us users of the technology and this appears to be a good entry into the fray.
Wayne Niemeyer of the McCrone Group showed some applications and results of "Low Voltage Scanning Electron Microscopy." Those of you who know the capabilities of these systems won't need "preaching to the choir" but I must say that it really is impressive what can be done with these systems. Wayne showed some beautiful images of exquisitely fine structural features, some in complex, deeply three-dimensional surfaces, all without any trace of charging, all in sharp, clear detail. I've seen 'em before but I was impressed again.
John Reffner of Nicolet/Spectra-Tech reminded us that "There is Microscopy in Infrared Microspectroscopy." He pointed out the complimentary nature of microscopy and microspectroscopy, that a scientist should be not a microscopist _or_ a spectroscopist, but both if he or she wants to fully exploit the capabilities of these instruments. It is appropriate that one from the company which actually emphasizes the importance of good microscopy (and good microscopes) as a part of microspectroscopy should make this point. Spectra-Tech is, in my opinion, to microspectroscopy what Zeiss used to be to microscopy. Yes, they may be the most expensive, but "you get what you pay for" here as elsewhere. I think that we should advocate retaining the capability to do good work with our equipment, even if we need to learn a bit more to take advantage of that capability, rather than accept instruments which have been "dumbed-down" to the least common denominator of the people who use them. So far, Spectra-Tech has not succumbed to any pressure they might feel to adopt that trend and I hope you'll join me in encouraging them to continue to hold the high ground. Someone needs to!
(Ok, ok, so I jamb in a bit of editorializing too. I'm not related to any of the companies I'm mentioning, so I think I can use a bit of license here.)
In the afternoon, Theodore M. Clarke of J.I. Case Corporation discussed "High Magnification Photomacrography Using the Kodak 1.6i/AB MegaPlus (TM) Camera." This provided a nice complement to the Polaroid talk, but this by an independant technical evaluator who put the Kodak product through some serious resolution tests, finding that it performed well if you accepted Ted's recommended 500 X NA rule of thumb for maximum useful magnification. This is 1/2 of the conventional wisdom but Ted made (and illustrated) a good argument for the more conservative standard. The Kodak product performed quite well in Ted's tests and I suggest all interested parties watch for the publication of his work in The Microscope in coming months. The technical detail of his work was too much to repeat here, but watch for the publication as it will be well worth using not only for its evaluation of the Kodak product but as a model for how digital cameras can be well and truely tested.
Also in the afternoon, Allen Whiteside of the McCrone Group presented two back-to-back papers on "Preparing Holey Carbon Films" and "AEM Preparations Using Holey Carbon Film." (Note that "The McCrone Group" is a different organization than the meeting hosts, the McCrone Research Institute. They separated years ago.) Once again, the folks at the McCrone Group demonstrate that they are masters of specimen manipulation (i.e., particle handling) and specimen preparation as well as analytical microscopy. I think they have probably originated more useful techniques than any other single organization. Allen showed their in-house technique for the preparation of holey carbon films which can and should be tailored to the specific needs of the analysis at hand. A film of one thickness, pore density, and web structure will be appropriate for one analysis, something different for another. They drop a solution of formvar in ethylene dichloride onto a glass slide which is rotated on a turntable to spin out the applied fluid while the solvent evaporates. The film is then lifted and applied to grids in a more-or-less conventional manner but the secrets of the technique lie in the speed of rotation and the formulation of the solvent/formvar solution. Again, the idea is to experiment and determine the best film preparation system for a particular kind of analysis, not adopt a single procedure. There is more, or course, and this was the subject of Allen's second talk. There are numerous techniques for applying the particles of interest to the holey film, again chosen to best suit the needs of the analysis.
I should close, but I just want to mention one more thing. There are many more papers being presented than I'm mentioning here. I'm only hitting on a few that seem particulary interesting to me for one odd reason or another. One should not infer anything from my failing to mention some of the other fine papers!
More to come, stay tuned!
Steve Shaffer MicroDataware
p.s. Yes, we are till publishing the Particle Atlas Electronic Edition (PAEE). Please contact me via EMail if you would like further information.
p.p.s. I know I'm a terrible speller and I'm preparing these things quickly while on the road and without access to my regular array of make-me-look-good software, so please forgive the rough nature of these communications. ;-)
I've dealt with both 35mm (immunofluorescence) and 3 1/4 X 4 1/4 inch (EM) negatives in both ways. Using a high quality light box and a very high quality macro lens on a fairly standard sort of CCD camera (Dage 72, 640X480, 8bits). I've gotten good digital images, printed on a Codonics dye-sub. But (my humble opinion) the quality doesn't approach what you can do in the darkroom. It is, however, WAY faster and easier. A fancier camera may help, but it all depends on what you want to do with the images. If the images are already on film, then I would tend to stick with silver grains over pixels. If you really need to go digital and have some money, scanners are probably the way to go for digitizing negatives -- check the list's archives, there has been much discussion of scanners.
Greg Martin Dept. of Cell Biology and Anatomy Johns Hopkins School of Medicine
On Tue, 22 Jul 1997, Mike Coviello wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hi All: } I recently put a message on the listserver requesting information on a } LogEtronics enlarger. Some responses I received suggested that I should } think about using a digital camera instead to capture images from negatives } and then process the images and then send them to a printer. I would like } some feedback from users. Is the quality close to that of film? Does anyone } have a system that they are extremely happy with? Are there any commercial } systems available? Please let me hear your experiences. } } Thanks, } } Michael Coviello } EM Lab Manager } The University of Texas -at- Arlington } Arlington, TX } E-mail coviello-at-mae.uta.edu } 817-272-5496 } } }
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Mike,
I would buy a Durst 1200 point source enlarger if you are doing high resolution electron microscopy ---- the logEtronics enlarger uses a scanning spot to equalize large contrast variations within an image and this enlarger prints like a diffuse source enlarger not a point source enlarger. We have found that lines are sharper and narrower with the point source enlarger. Our work requires darkroom printing of reversal negatives made from thin vertically Pt-C replicated specimens in which we want to visualize molecular details on the structural highlights. Usually these features only become visible after printing the reversal negative on fiber based paper and drying the print on glossy paper---- For TEM publications that do not need this detail as part of their story, it is easy to scan in images at a hardware resolution of 600 dpi and label the image with Adobe photoshop---- I find that I need access to a good darkroom as well as the appropriate digital darkroom equipment.
George Ruben Dept. Biological Sciences Dartmouth College Hanover, NH 03755
Hi everyone, for many years I have been embedding in Spurr's resin but have recently decided to give glycol methacrylate a whirl for thicker LM sections of plant material. Many years have passed since my last work with GMA. I remember that we made a plexigalss container which we could evacuate the oxygen from by flushing with nitrogen and that polymerization was then carried out below a longwave UV light placed in the container. Does this sound correct? If so does anyone have any plans or tricks that might assist me in constructing such a device? Cheers, John
================= C. John Runions, Ph.D Section of Ecology and Systematics Corson Hall Cornell University Ithaca, New York USA 14853
We've gone digital TEM and everyone loves it for ease of capture and speediness to the investigator. Our 2Kx2K images look great on a large monitor and printed with on a Tektronix dye-sub printer. Heck, even an Epson Stylus printer with 1440 dpi makes darn good prints (on photo-quality paper)! In a side-by-side comparison, I'll take a high contrast print from our Durst EM1200 over the dye-sub. However, dollar for dollar, one alternative to consider is to transfer TEM negatives to Kodak PhotoCD and print only select images, when you need publication quality, to the Fujix Pictrograph 3000 digital printer (which costs less than the Durst with all the electronic bells and whistles.) Thereby you have your negative, a high-resolution digital image, a digital print, and you can still make a print if you really have to.
There's my humble opinion. Thank you.
Walter F. Bobrowski Subcellular Pathology Parke-Davis Pharmaceutical Research Ann Arbor, MI 48105
To all who would advocate a totally digital darkroom,
While on the whole I have to agree that the direction that science/microscopy is taking is a digital one, I must point out that there are some serious shortcomings to digital imaging. The most serious, in my mind, is the issue of archivability. While I could launch into this myself, I'd prefer to bring to everyone's attention the following article (which addresses this issue far better than I can):
"Ensuring the Longevity of Digital Documents" Jeff Rothenberg, Scientific American, January 1995
If we can't honestly answer the question of how we, or anyone else, will be able to use (or even access) our digital images 30 years from now, we need to re-evaluate the speed with which we are going digital. If properly cared for, film can last for decades. The uncertain aging of digital storage media, the often rapid obsolescence of drive mechanisms & media types and the ongoing changes in image file formats are cause for concern.
Yours, Doug Cromey ..................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Sr. Research Specialist 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
Our multi-user facility is currently archiving our confocal and LM digital images on Panasonic optical disks (re-writable, very stable -at- about $125 for 1 GB). The disadvantage is that few of our users have their own Panasonic drives so most people simply archive the images at our core and then move the ones they want by FTP as needed. I would like to switch to a more universal medium - namely CD ROM's. My understanding is that CD's can now be written to in multiple sessions so you don't need to fill an entire disk at once. Furthermore, it is my understanding that a disk of TIFF images should be readable by both IBM/WINTEL and Mac/PowerPC types computers. Is anybody actually doing this? Comments on how reliable are the recorders, which ones are best, pitfalls, etc would be appreciated. Before I get a dozen advocates of ZIP/Jazz drives, I don't want to go that route since that they are not as ubiquitous as CD drives. Thanks in advance.
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
Mike, your needs for image quality may exceed ours (industrial research laboratory, not as interested in publishing outside as academic), but we recently removed all of our darkrooms because we had not processed a negative or print in two years.
All of our microscopy (AFM, TEM, SEM, and Optical) is done with digital image capture. The speed to get a hardcopy, the flexibility of sharing images worldwide via electronic transfer, and the cost per print/image all strongly favor total digital imaging. Although the quality of a typical digital image does not equal that which can be obtained with photographic film they meet our needs completely. For the highest quality digital imaging in optical microscopy we use a Kodak 460 digital camera or a Leaf MicroLumina (3500 X 2400 pixels). Storage of images is done on CD-ROM for archival capability and erasable Magneto-Optical CD (40GB jukebox on a network server) is used for day-to-day image storage. Photoshop is used for image "retouching". Reports are composed in MicroSoft Word and hardcopy of images are obtained with networked Kodak 8650 printers (one dedicated to B/W and one for color).
As a final note, we've been using video and digital images for 9 years and would never think of going back to a photographic imaging process.
********************************************************* Dr. Dennis B. Barr Research Laboratories Eastman Chemical Company Kingsport, TN 37664-5150 Phone:423-229-2188 Fax:423-229-4558 Email: dennbarr-at-eastman.com ********************************************************
} -----Original Message----- } From: Mike Coviello [SMTP:Coviello-at-mae.uta.edu] } Sent: Tuesday, July 22, 1997 4:24 PM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Digital Darkrooms } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
If you spend $30,000--$50,000 for hardware and software (which will be obsolete in less than five years) you can produce very good hardcopy images comparable to conventional photography. The digital process requires a highly skilled system caretaker and the learning period for neophytes is much longer. For every year that you can delay the transition to digital you will reduce the problems significantly. These statements are based on my experiences with photography for 32 years and digital graphics for 10 years. Larry D. Ackerman (415) 476-8751 Howard Hughes Medical Institute FAX (415) 476-5774 UCSF, Box 0724, Rm U426 533 Parnassus Ave. mishot-at-itsa.ucsf.edu San Francisco, CA 94143
On the other hand... I can send digital images to any of my JIT-based manufacturing customers in a matter of moments via e-mail. Do that with film! I can store images with and without annotation in an inexpensive database that is continually being upgraded by its developers... and is forward and backward compatible with our standard desktop applications. I have reduced my Polaroid film expenditures by at least five thousand dollars per year (with the attendant costs to order, deliver, bill, etc.) My image acquisition system is never out of stock. Images don't get "lost in the mail" I can extract a variety of numbers from a digital image easier than using primitive tools like an ASTM grain size overlay.
It should be noted that the medical industry is a driving force for improvements in digital imaging technologies and electronic collaboration precisely because of the failure of film to fulfill the needs of today's customers.
} ------------------------------------------------ } Opinions or statements expressed herein, rational or otherwise, do not } necessarily reflect those of my employer. } } Harold J. Crossman } OSRAM SYLVANIA INC. } Lighting Research Center } 71 Cherry Hill Dr. } Beverly, MA 01915 } Phone: (508) 750-1717 } E-mail: crossman-at-osi.sylvania.com } } Our web sites: www.sylvania.com } www.siemens.com } -- } } "Crossman, Harold" {crossman-at-osi.SYLVANIA.com}
We use CD-ROMs for archiving images, data, etc. If you make the CD-ROM in ISO-9660 format it can be read by PC's, Macs, and Unix machines. ISO-9660 does not allow long file names. There are other formats - Joliet system - that allow long file names but these can only be read in Win95 machines. I recommend that you have a dedicated machine for making CD-ROMS. Partition the hard drive so that your system files are on C-drive and leave the D-drive for files to be archived. Two of the major manufacturs are Yamaha and Pinnacle Micro. If you look up their web-sites and read the FAQ's related to installation and troubleshooting, you will get some idea of the important issues in setting up a system (There are certain hard drive specifications, etc.) Multisession is possible, but you need software that will read a multisession disk (usually the software package that was used to make the CD). If you make a multisession disk, and place it in a computer without the proper software - the computer will only see the last session. This drawback may be changing (changed?) with the next generation of machines and software. Overall, I think CD-ROM is the current best method for archiving data.
Regards,
John J. Turek, Ph.D. Associate Professor Director, Electron Microscopy Laboratory and Core Laboratory for Image Analysis and Multidimensional Applications (CRISTAL) Department of Basic Medical Sciences 1246 Lynn Hall, G193C Purdue University W. Lafayette, IN 47907-1246 Phone: 765-494-5854 Fax: 765-494-0781 Email: jjt-at-vet.purdue.edu
We have installed writable CD-ROM on a UNIX system using software manufactured by Microson, called GEAR 32.
There have been serious problems arising from incompatibility of UNIX with CD-ROM technology (asynchronous versus synchronous - The Unix machines deliver information when they are ready but the CD writing process requires information at a constant rate which is determined by the disc speed). I don't believe this is a problem with PC's, but watch out if you want to use CD-R on a UNIX platform. We had to buy a new external hard drive, directly connected to the CD-R device, in order to get everything to work reliably.
Also, and this may apply to you as well, the claims of the software manual concerning various options like multisession backing up were not actually implementable. We must write an entire CD at once. This is is not so bad actually as we deal with large quantities of image data, and the disks themselves are now only around $4-$5, which you really can't beat for 650MB of space. The software packages available when working in the PC world may be better, but beware, and make sure you get what is advertised. I think this technology has a future and that the microscopy world can benifit. However, the software for running it, at least on a UNIX platform, has some progress still ahead of it.
Wharton
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Wharton Sinkler PhD Department of Materials Science and Engineering Northwestern University 2225 North Campus Drive Evanston, IL 60208-3108 tel: (847) 491-7809 fax: (847) 491-7820 email: sinkler-at-apollo.numis.nwu.edu
On Wed, 23 Jul 1997, Tom Phillips wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Our multi-user facility is currently archiving our confocal and LM digital } images on Panasonic optical disks (re-writable, very stable -at- about $125 } for 1 GB). The disadvantage is that few of our users have their own } Panasonic drives so most people simply archive the images at our core and } then move the ones they want by FTP as needed. I would like to switch to a } more universal medium - namely CD ROM's. My understanding is that CD's can } now be written to in multiple sessions so you don't need to fill an entire } disk at once. Furthermore, it is my understanding that a disk of TIFF } images should be readable by both IBM/WINTEL and Mac/PowerPC types } computers. Is anybody actually doing this? Comments on how reliable are } the recorders, which ones are best, pitfalls, etc would be appreciated. } Before I get a dozen advocates of ZIP/Jazz drives, I don't want to go that } route since that they are not as ubiquitous as CD drives. Thanks in } advance. } } } Thomas E. Phillips, Ph.D. } Associate Professor of Biological Sciences } Director, Molecular Cytology Core Facility } 3 Tucker Hall } University of Missouri } Columbia, MO 65211 } (573)-882-4712 (voice) } (573)-882-0123 (fax) } } }
We have installed writable CD-ROM on a UNIX system using software manufactured by Microson, called GEAR 32.
There have been serious problems arising from incompatibility of UNIX with CD-ROM technology (asynchronous versus synchronous - The Unix machines deliver information when they are ready but the CD writing process requires information at a constant rate which is determined by the disc speed). I don't believe this is a problem with PC's, but watch out if you want to use CD-R on a UNIX platform. We had to buy a new external hard drive, directly connected to the CD-R device, in order to get everything to work reliably.
Also, and this may apply to you as well, the claims of the software manual concerning various options like multisession backing up were not actually implementable. We must write an entire CD at once. This is is not so bad actually as we deal with large quantities of image data, and the disks themselves are now only around $4-$5, which you really can't beat for 650MB of space. The software packages available when working in the PC world may be better, but beware, and make sure you get what is advertised. I think this technology has a future and that the microscopy world can benifit. However, the software for running it, at least on a UNIX platform, has some progress still ahead of it.
Wharton
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Wharton Sinkler PhD Department of Materials Science and Engineering Northwestern University 2225 North Campus Drive Evanston, IL 60208-3108 tel: (847) 491-7809 fax: (847) 491-7820 email: sinkler-at-apollo.numis.nwu.edu
On Wed, 23 Jul 1997, Tom Phillips wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Our multi-user facility is currently archiving our confocal and LM digital } images on Panasonic optical disks (re-writable, very stable -at- about $125 } for 1 GB). The disadvantage is that few of our users have their own } Panasonic drives so most people simply archive the images at our core and } then move the ones they want by FTP as needed. I would like to switch to a } more universal medium - namely CD ROM's. My understanding is that CD's can } now be written to in multiple sessions so you don't need to fill an entire } disk at once. Furthermore, it is my understanding that a disk of TIFF } images should be readable by both IBM/WINTEL and Mac/PowerPC types } computers. Is anybody actually doing this? Comments on how reliable are } the recorders, which ones are best, pitfalls, etc would be appreciated. } Before I get a dozen advocates of ZIP/Jazz drives, I don't want to go that } route since that they are not as ubiquitous as CD drives. Thanks in } advance. } } } Thomas E. Phillips, Ph.D. } Associate Professor of Biological Sciences } Director, Molecular Cytology Core Facility } 3 Tucker Hall } University of Missouri } Columbia, MO 65211 } (573)-882-4712 (voice) } (573)-882-0123 (fax) } } }
} ... } } . . . The most serious, in my mind, is the issue of archivability. } ... } } "Ensuring the Longevity of Digital Documents" } Jeff Rothenberg, Scientific American, January 1995 } } If we can't honestly answer the question of how we, or anyone else, } will be } able to use (or even access) our digital images 30 years from now, we } need } to re-evaluate the speed with which we are going digital. If properly } } cared for, film can last for decades. The uncertain aging of digital } storage media, the often rapid obsolescence of drive mechanisms & } media } types and the ongoing changes in image file formats are cause for } concern. } } ...
I agree ... however, while there exists good reasoning to avoid magnetic media for long-term archiving, there seems to be little concern in this regard for magneto-optical or CD ROM media. Also regarding archival quality ... digital hardcopy will not hold up to long-term storage of preperly washed photographic papers (... I think it has already been mentioned that the resolution and the quality of the grayscale for even the best digital printers doesn't even come close to chemica darkroom printing ...). Still ... even as an ex-photographic art student ... I can appreciate the new capabilities (in general) that a digital darkroom has over the traditional for producing *lots of work* and its being "publication quality" ... if not "art for the purist" quality ...
cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
Hi Tom. I have been burning CD's for our SEM users for about a year and a half now. We have an HP 4020i and I am using the Easy CD software package (supposedly the best). Of the 50 or so users who have archived onto CD only 2 have not been able to get images I wrote and another says he can't but I think in his case it is the operator. Both of the users who are having trouble are able to read the data from the first write, but not from successive write sessions. Thay have brought their disks back into the lab and I am able to read ALL the data from ALL write sessions on each of our four cd readers. Our computer guy here asked how old the drives they were using to read the disks were cause if they are too old they may not support the multi-session standards. Only one user seems to have such an aged cd rom. The other is baffeling. Heck, my cd-rom at the house is old as the hills (and the cheapest I could find) and it does fine. HP was no help in this matter either so if anyone has any ideas I would sure appreciate hearing from you. I can still say I reccommend the writer and feel it is probably the best deal. I don't feel bad making the users buy an $8-9 disk, even the guys who can only read the first session. You are correct that both MAC & PC's will read the files as long as the MAC is running system 7 or better. SGI will not nor will OS2. Good luck
At 10:32 AM 7/23/97 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 352-392-1295 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
We have been archiving to CDs with a Philips 2000 for almost two years with no problems. Much cheaper than opticals. $9.00 for 600 MB.
Bob Morphology Core Seattle
On Wed, 23 Jul 1997, Tom Phillips wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Our multi-user facility is currently archiving our confocal and LM digital } images on Panasonic optical disks (re-writable, very stable -at- about $125 } for 1 GB). The disadvantage is that few of our users have their own } Panasonic drives so most people simply archive the images at our core and } then move the ones they want by FTP as needed. I would like to switch to a } more universal medium - namely CD ROM's. My understanding is that CD's can } now be written to in multiple sessions so you don't need to fill an entire } disk at once. Furthermore, it is my understanding that a disk of TIFF } images should be readable by both IBM/WINTEL and Mac/PowerPC types } computers. Is anybody actually doing this? Comments on how reliable are } the recorders, which ones are best, pitfalls, etc would be appreciated. } Before I get a dozen advocates of ZIP/Jazz drives, I don't want to go that } route since that they are not as ubiquitous as CD drives. Thanks in } advance. } } } Thomas E. Phillips, Ph.D. } Associate Professor of Biological Sciences } Director, Molecular Cytology Core Facility } 3 Tucker Hall } University of Missouri } Columbia, MO 65211 } (573)-882-4712 (voice) } (573)-882-0123 (fax) } } }
} ... } } } } Our multi-user facility is currently archiving our confocal and LM } digital } } images on Panasonic optical disks (re-writable, very stable -at- about } $125 } } for 1 GB). The disadvantage is that few of our users have their own } } } Panasonic drives so most people simply archive the images at our } core and } } then move the ones they want by FTP as needed. I would like to } switch to a } } more universal medium - namely CD ROM's. My understanding is that } CD's can } } now be written to in multiple sessions so you don't need to fill an } entire } } disk at once. Furthermore, it is my understanding that a disk of } TIFF } } images should be readable by both IBM/WINTEL and Mac/PowerPC types } } computers. Is anybody actually doing this? Comments on how } reliable are } } the recorders, which ones are best, pitfalls, etc would be } appreciated. } } Before I get a dozen advocates of ZIP/Jazz drives, I don't want to } go that } } route since that they are not as ubiquitous as CD drives. Thanks } in } } advance. ...
Whereas I would have thought CDROM should be the best method (... considering the cost of the media ...), judging from the responses we've seen I'm glad I went with a Fujitsu 640 magneto-optical. It has a solid SCSI interface (... I use Adaptec ...) and it is re-writable. I had some early issues when it first came out (... we had been using 230Mb for 1.5 years ...) but it works flawlessly now. The cost per 640Mb cartridge is (however) $35 ... and (... of course ...) the drives aren't as ubiquitous. We use it in a intranet configuration, and all image files and data from the SEM and Cameca archive directly to it. Non-WIntel users download from it via "anonymous" FTP ...
... another point of view ...
cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
If some of you EDS practioners (yes, I'm a WDS practioner) could help me with this I would be most appreciative:
1. presently I am involved in a project for which I have to examine zinc arsenides - the process by which these are made can, at times, also produce elemental Zn, ZnO and elemental As (yuk!, and then some)
2. I mainly characterize these beasties by x-ray diffraction, but in this particular inspection I am doing some SEM work too
3. the samples I am looking at are suspected to be mainly the zinc arsenide phase(s), however, I get EDS spectra with highly variable Zn/As - in fact, more wide-ranging than expected for various of the possible zinc arsenides
4. I suspect that the Zn/As variation has as much to do with loss (diminishment) of the As signal (due to absorption?) as it does with variation in the ZnAs
5. to that end (#4), I performed a test yesterday in which I examined just one crystal type (based on morphology) in areas of the sample where these crystals were at the "surface" of the sample and at various "depths" within holes and/or depressions.....basically, the deeper in the "hole" the lower the As signal
6. concurrent with the dimished As, I also observed diminishment of Zn L-alpha line (also a low-energy x-ray) such that Zn-K/Zn-L was also highly variable - would you consider the latter a further indication of absorption?
In the interim as, I await your response, I intend to run x-ray diffraction on the samples with "apparently" low As (some are even As "absent") to "confirm" whether or not the Zn or ZnO phases could be present. Together (my XRD and preliminary SEM, plus your sage advice) I hope to resolve this.
Thanks, in advance!!
Winton
Dr. Winton Cornell Senior Research Associate & Supervisor, Microanalysis Laboratory Department of Geosciences The University of Tulsa 600 South College Tulsa, OK 74104-3189
Sharon et.al. That knife maker is an everlasting instrument but the adjustments must be right, otherwise its a big waste of glass and time. 1 Adjust the back holder of the glass, so that the cutting stroke across the diagonal of the glass square stops one or two mm before running off the glass. 2 Loosen the front holder and press the moving part to just touch the corner of the glass. Increase the push against the spring tension by two scale divisions and then tighten the knurled knob. 3 The front and back lateral adjustments must now be set. Centre both of these, check with a glass square inserted and a piece of paper as a straight edge that the centre line would be corner to corner on the glass. 4 Now adjust the back lateral adjustment so that the centre line would be 1 to 2mm to the left of the back corner. Tighten the lock-nut. 5 Repeat for the front adjustment but to the right of the front corner. 6 Break a couple of test squares, without the front damper, which pushes a bit of rubber against the glass, applied. Those knives should have their edges close, but not across the corners. If required adjust the lateral controls. 7 Once well set, the laterals never need adjustment. Draw a marker pen line across the dials to indicate settings or tighten them just beyond finger strengths.
Never touch the clamping head when it is lowered; it determines the clamping pressure and over-tightening its locking device is pointless.
The rubber damping device slows down the moment of fracture of the front knife only. This result in a wider stress free area for that knife. Apply the damper after clamping the head. Mover rubber front to just touch glass plus two scale divisions. The glass should not move back. Score, break and then, always before lifting the clamping head, release the damper.
I am writing this from memory but I've taught that operation a few hundred times. Ask me if you still have problems with the knife maker. Disclaimer: ProSciTech and most other EM suppliers supply microtomy glass. We should have an interest in maladjusted knife-makers. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 400+ Links, MSDS ************************ http://www.proscitech.com.au
---------- } From: Sharon C. Thomas {sthomas-at-lanl.gov} } To: Microscopy-at-sparc5.microscopy.com } Subject: Help requested for maladjusted glass knifemaker } Date: Wednesday, 23 July 1997 3:44 } The knifemaker available to me is a LKB Type 7801A from LKB Instruments Inc. } out of Rockville Maryland, but the company has since been taken over by } Leica/Leo. The knifemaker has two wheel type gauges that adjust the tension } from opposing sides on the glass piece. I can make glass squares, but when } I try to cut the square into the two knives, I don't get the correct } shapes, even } though I have tried systematically changing the settings on the two gauges } many different ways. Typically, the "sharp" edge may be chipped, the } reflection line in the glass is going in the opposite direction to what it } should } and the opposite edge to the sharp edge may be either sharp as well or too } thick a blunt edge. That's just one of the two knives; its' counterpart } often has } two "sharp" edges. Can anyone with any direct experience with this model of } knifemaker provide me with any advice on how to adjust the settings to produce } two glass knives? } } Thanks in advance for any assistance that can be provided. } }
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Tom:
While we have do many Zip drives in use in our multi-user facility, they are used for purposes other than archive storage. We archive all of our digital images (from 6 major instruments) onto CD-ROMS. We have more than 60 Gbytes of active, primary storage on on-line servers, which is good enough for 3-4 months of image storage, after which the oldest images are downloaded onto CDs. This has worked nicely for us for the last 3-4 years. We store all our images in their original formats (DigitalMicrograph, Adobe PhotoShop etc.) so they can be handled ultimately just as if they had just been recorded.
Hope this helps.
Larry
} } Our multi-user facility is currently archiving our confocal and LM digital } images on Panasonic optical disks (re-writable, very stable -at- about $125 } for 1 GB). The disadvantage is that few of our users have their own } Panasonic drives so most people simply archive the images at our core and } then move the ones they want by FTP as needed. I would like to switch to a } more universal medium - namely CD ROM's. My understanding is that CD's can } now be written to in multiple sessions so you don't need to fill an entire } disk at once. Furthermore, it is my understanding that a disk of TIFF } images should be readable by both IBM/WINTEL and Mac/PowerPC types } computers. Is anybody actually doing this? Comments on how reliable are } the recorders, which ones are best, pitfalls, etc would be appreciated. } Before I get a dozen advocates of ZIP/Jazz drives, I don't want to go that } route since that they are not as ubiquitous as CD drives. Thanks in } advance. } } } Thomas E. Phillips, Ph.D. } Associate Professor of Biological Sciences } Director, Molecular Cytology Core Facility } 3 Tucker Hall } University of Missouri } Columbia, MO 65211 } (573)-882-4712 (voice) } (573)-882-0123 (fax)
Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
I am a biophysics graduate student. One of my exam question is how to correct CTF in image reconstruction. After finishing the exam, I have more questions than answers!
What kinds of contrast exists in electron microscopic image for weak-phase object? Is amplitude contrast the same as aperture contrast?
What is the contribution of inelastic scattering on the image contrast? How to account for it?
How does partial spatial coherence and temporal coherence influence the image and electron diffraction?
Is it always sufficient to use just first order approximation ( the linear theory of phase and amplitude contrast image formation ) for CTF consideration?
Is it true for low spatial frequencies ignoring the CTF was better than compensating for phase contrast alone?
I guess compensation for the CTF was necessary and sufficient to accurately reconstruct molecular densities. Could anyone tell me the current ways for accurate determination of CTF?
By improving the different components in CTF, what is the optimal contrast could be achieved in image?
Is it possible to put EM reconstruction on the same scale with the structures derived from X-ray crystallography and NMR after deliberate correction of CTF, solvent effects and differences between atomic scattering factors for electron and X-ray?
To what extend we could use the insights obtained by building models and comparison of the models with EM reconstruction?
Answers to any parts or aspects of these confusions I have will be greatly appreciated!
I am a biophysics graduate student. One of my exam question is how to correct CTF in image reconstruction. After finishing the exam, I have more questions than answers!
What kinds of contrast exists in electron microscopic image for weak-phase object? Is amplitude contrast the same as aperture contrast?
What is the contribution of inelastic scattering on the image contrast? How to account for it?
How does partial spatial coherence and temporal coherence influence the image and electron diffraction?
Is it always sufficient to use just first order approximation ( the linear theory of phase and amplitude contrast image formation ) for CTF consideration?
Is it true for low spatial frequencies ignoring the CTF was better than compensating for phase contrast alone?
I guess compensation for the CTF was necessary and sufficient to accurately reconstruct molecular densities. Could anyone tell me the current ways for accurate determination of CTF?
By improving the different components in CTF, what is the optimal contrast could be achieved in image?
Is it possible to put EM reconstruction on the same scale with the structures derived from X-ray crystallography and NMR after deliberate correction of CTF, solvent effects and differences between atomic scattering factors for electron and X-ray?
To what extend we could use the insights obtained by building models and comparison of the models with EM reconstruction?
Answers to any parts or aspects of these confusions I have will be greatly appreciated!
} I am a biophysics graduate student. One of my exam question } is how to correct CTF in image reconstruction. After finishing } the exam, I have more questions than answers! } (snip - long list of questions deleted...)
You ask several good questions that space (and time) does not permit me to answer in detail. May I suggest that you check out the Ph. D. Thesis of Xiadong Zou (Electron Crystallography of Inorganic Structures - Theory and Practice.) This was published in the Chemical Communications of Stockholm University (1995 No. 5.) You may be able to get a copy from your library. If not, write the Department of Structural Chemistry, Arrhenius Laboratory, Stockholm University, S-106 91 Stockholm, Sweden. Xiadong's thesis advisor was Professor Sven Hovmoller.
In the first part of her thesis, Xiadong does a great job of summarizing the theory of electron imaging and diffraction in a unified fashion. One of the problems that we all face is that the theory was developed by several communities and the terminology and notation is often conflicting and confusing. For example, one needs to be careful to decide whether the author chooses underfocus to be negative or positive. Xiadong shows several examples of the effect of correction for CTF on image reconstruction.
-- Best Regards, John Minter
Eastman Kodak Company Phone: (716) 722-3407 Analytical Technology Division FAX: (716) 477-3029 Room 2112 Bldg 49 Kodak Park Site email: minter-at-kodak.com Rochester, NY 14562-3712 calendar: via PROFS
Haven't yet convinced the guys at work to spend the whopping {g} $400-600 to implement a CD-R, but do have one on the home system which works great for back-up/archiving. Is SCSI2 (aren't they all?) running EzCD Pro software.
My HP 6020 does not support "packet mode" so it can't be treated like a floppy, but does support "multi-session". It is not necessary to write "disk at once". The caveat is that for each writing "session" something like 13 to 20 Mb of CD space is consumed as "overhead". ...But at $6 for 650Mb, do we care?
As to compatibility... some old CD players (under W3.1?) read the first session and quit. When those systems were made, multisession was not commonly available.... I haven't yet found a system old enough that it would not read the CDs I burned.
Who really knows how long the CD-Rs will last (and will the hardware be around to read them?). TDK rates their disks for 100 years....
If your only goal is to get thicker, larger sections with GMA, I have a proceedure that doesn't require a special chamber or UV light. I have used this procedure for 15 years with very good results. The blocks stain well with toluidine blue, but little else, though. I use a modified embedding procedure for JB-4, developed by Ann Klinker and Marge Hukee 15 years ago. It does require the blocks to be set without air, we use parafilm sealed to the top of the mold. E-mail me if you are interested in the details. kna101 utdallas.edu
Karen Pawlowski
On Wed, 23 Jul 1997, C. John Runions wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hi everyone, for many years I have been embedding in Spurr's resin but } have recently decided to give glycol methacrylate a whirl for thicker LM } sections of plant material. Many years have passed since my last work with } GMA. I remember that we made a plexigalss container which we could } evacuate the oxygen from by flushing with nitrogen and that polymerization } was then carried out below a longwave UV light placed in the container. } Does this sound correct? If so does anyone have any plans or tricks that } might assist me in constructing such a device? Cheers, John } } ================= } C. John Runions, Ph.D } Section of Ecology and Systematics } Corson Hall } Cornell University } Ithaca, New York } USA 14853 } } email cjr14-at-cornell.edu [ie. cjr(fourteen)-at-...] } phone (607) 254-4282 } Fax (607) 255-8088 } } }
John Hunt writes: } From: John Hunt {hunt-at-msc.cornell.edu} } Subject: Re: CD-ROM's for archiving - any experience? (fwd) } To: rick-at-warf.msc.cornell.edu (Rick Cochran) } Date: Thu, 24 Jul 1997 09:29:12 -0400 (EDT) } } I am forwarding this exchange from the microscopy listserver for } your interest.
Which OS is best to run your CDR under depends on what environment your date is being acquired in. The three environments which pop to mind are Unix, Windows, and Mac. CDR solutions are available for all three environments.
For our Unix environment, we have recently succeeded in setting up CDR under Linux using freeware utilities. You need a SCSI system with a suitably large disk, 'mkisofs' for mastering the CD images, and 'cdwrite' or 'cdrecord' for actually writing the CD. The only problem we ran into was that most CDR drives require the 'SCSI disconnect' feature which was disabled by default in the Linux SCSI driver for our SCSI adapter.
mkisofs is available at tsx-11.mit.edu:/pub/linux/packages/mkisofs
cdwrite and cdrecord are available at sunsite.unc.edu:/pub/Linux/utils/disk-management
Since there is no industry standard for the SCSI commands for CDR drives, support for each drive must be explicitly written into the burning utility. You should determine which drive cdwrite and cdrecord support before purchasing a drive.
-- |Rick Cochran phone: 607-255-7223| |Cornell Materials Science Center FAX: 607-255-3957| |E20 Clark Hall, Ithaca, N.Y. 14853 email: rick-at-msc.cornell.edu| | "The Founding Fathers did not establish the United States as a | | democratic republic so that elected officials would decide trivia, | | while all great questions would be decided by the judiciary." | | Judge Andrew Kleinfeld |
At 11:24 PM 7/23/97 -0400, you wrote: } My question is: Has anyone *ever* gone back to access and use original } image data that is 30 years old? I personally do not remember ever using } original images that were even 5 years old....
Actually, I used to work in a Pathology diagnostic EM lab and we certainly went back 20 years sometimes (rare diseases often require an entire career to accumulate enough examples to publish about). I imaging the need for archiving images is different for each field, but I still think its a bit short sighted to not consider the future while living in the present. Although this is off the science track, I wonder how our grandkids will manage to view our digital snapshots when we're pretty sure that dye-sub prints don't last that long and they need to find a viewer/reader for our family album CD-ROM disk.
} Perhaps we should realistically not place too great an emphasis on the } usefulness of saving all images for 30 years. CD-ROMs supposedly have that } capability, but who believes that today's CDs will be readable by any } technology available in 30 years? Anybody keep their old 8-track tape } players in good service? I think most digital image formats will have to } periodically be upgraded to the latest storage technologies, just as people } take 16 mm home movies and convert them over to video tapes. Of course, } this does not have to be done with film, and negatives probably will always } be able to be converted easily to hard copy many years in the future....but } I still never make images on film any more...
I may not have an 8-track (anymore), but how could we have the current crop of re-releases of old musical works (originally recorded as analog) if the music industry hadn't had a long lasting standard that they could still work with today? Remember too that movie to video transfer results in a lower resolution image (loss of information) stored on a tape format (I hope you meant VHS, not the now obsolete BetaMax) that probably has a 5 year lifetime.
Actually, I'm of the mindset that of the mass storage technology currently available today, CD-ROMs are the only ones likely to be readable in 30 years. That "Amazing Kreskin" like prediction (opinion) is based primarily on the large market presence of CD readers (one industry analyst believes that the installed base of CD devices will reach 150 million by the end of 1997, source: Advanced Imaging Magazine), compared with almost everything else.
Don't get me wrong, I like digital imaging for most of the same reasons everyone else likes it. I use it here at work and I try to teach students and staff about it because I think its the way science is going. I'm just concerned that we, as microscopists, are headed into this without realising all the issues related to archiving of and longevity of images.
Yours, Doug ..................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Sr. Research Specialist 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
Damian Neuberger mentioned that he had been told that ZIP drives were being discontinued. I think his source of information must have been wrong. Iomega recently introduced an internal SCSI version in addition to already existing external parallel port and external SCSI. A number of computer companies are now offering ZIP drives as standard equipment. We have used ZIP drives for several years on this campus for storing images from TEMs, SEMs, AFM/STMs, and Laser Confocal microscopes and we have been extremely pleased with them. The disks are sold by our local Best Buy Store as well as at most computer stores and Iomega has licensed other manufactures to produce the disks. We also have the option of storing images on a Panasonic Read/Write Magneto-Optical drive (1 GigaByte disks)on our SEM, but most users prefer the ZIP. I for one don't like the idea of having a thousand or so images on one disk. I much prefer to have several ZIP disks with a hundred or so images on each. Stanley L. Flegler, Assistant Director Center for Electron Optics Michigan State University
I can add little to what Jim Darley has said about setting up the knifemaker but I was wondering whether you used to get good knives and now get bad or have just started using the LKB.
Do you have the instruction charts for the knifemaker? Our LKB 7801A has a glossy card 4 page manual and a condensed instruction laminated chart which are very useful, when you understand them. They are normally hidden under the machine on a little purpose built tray.
When you say that the reflection line is the wrong way do you mean that the 'meniscus' in the glass curves the wrong way? This might depend on which way up you score your rhomboid/squares to make the triangles. We make 50 degree knives from 100/80 degree rhomboids by first producing the rhomboids by scoring on the roughened edge side of the glass strips. Then to bisect the rhomboid we turn it over so all the rough edges are at the bottom. (I hope you follow my meaning).
Finally what's your glass like - it hasn't been dropped or damaged. Is it the right stuff for e.m. - we use 6mm thick glass from a reputable e.m. supplier.
I hope this is of help but I'm sure that if you combine all of the replies you will solve your problem.
Malcolm Haswell e.m. unit University of Sunderland U.K. ----------
Sharon et.al. That knife maker is an everlasting instrument but the adjustments must be right, otherwise its a big waste of glass and time. {SNIP} ---------------------------------------------------------------------------- ------------------------- } From: Sharon C. Thomas {sthomas-at-lanl.gov} } To: Microscopy-at-sparc5.microscopy.com } Subject: Help requested for maladjusted glass knifemaker } Date: Wednesday, 23 July 1997 3:44 } The knifemaker available to me is a LKB Type 7801A from LKB Instruments Inc. } out of Rockville Maryland, but the company has since been taken over by } Leica/Leo. The knifemaker has two wheel type gauges that adjust the tension } from opposing sides on the glass piece. I can make glass squares, but when } I try to cut the square into the two knives, I don't get the correct shapes, even } though I have tried systematically changing the settings on the two gauges } many different ways. Typically, the "sharp" edge may be chipped, the } reflection line in the glass is going in the opposite direction to what it should } and the opposite edge to the sharp edge may be either sharp as well or too } thick a blunt edge. That's just one of the two knives; its' counterpart often has } two "sharp" edges. Can anyone with any direct experience with this model of } knifemaker provide me with any advice on how to adjust the settings to produce } two glass knives? } } Thanks in advance for any assistance that can be provided.
I got a message from G. Arentieri asking for help. There was no return address, so I cannot address him directly. I can do a discussion of long term storage of tissue, but it is not of general interest to the group, so I would like to send it to G.Arentieri directly. Greg, Could you try and contact me again? This time type your e-mail address just in case of another snafu. Bye, Hildy
I have been transfecting mammalian cells with a dopamine receptor and trying to detect, via fluorecence microscopy, its expression with a polycolonal antibody that I made to one of this dopamine receptor's intracellular loops. (The antibody detects the dopamine receptor epitope, to which the antibody was made, on western blots, when expressed by E. coli in the context of a fusion protein, so I know the antibody does recognize the epitope against which it was made.)
When I express the receptor in transfected HEK293 cells, fix the cells by methanol:acetone, and do fluorescence microscopy, I get no detection of receptor expression. I engineered a " FLAG tag" into the receptor and stained for FLAG labeling;the FLAG epiope gave intense staining, so I know the dopamine receptor is being expressed.
Does anyone have any thoughts on what factors I might to alter in my immunofluorescence assay in order to make the dopamine receptor antibody recognize the dopamine receptor in the transfected cells? It seems as if the receptor is folded in a conformation such that the epitope that the antibody recognizes is buried / not exposed. Are there better/more appropriate fixation methods? Does anyone have information on live staining or microwave heating/steaming of the cells?
Hello there, MAS Members and other interested parties, The latest issue of The Microbeam Analysis Society Newsletter, MicroNews (Summer 97), is now available on the Web at:
John Mansfield North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (313) 936-3352 FAX (313) 936-3352 Cellular Phone: (313) 715-2510 (Leaving a phone message at 936-3352 is preferable to 715-2510) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html
Dear all, I need some information. I have a Hummer VI-a sputter coater and I am having trouble finding where to get targets. EMCORP which sold the instrument seems to be out of business. Their phone numbers do not work. Any help will be appreciated.
TIA Greg Rudomen University microscopy Imaging Center S.U.N.Y. -at- Stony Brook
I got a message from G. Arentieri asking for help. There was no return address, so I cannot address him directly. I can do a discussion of long term storage of tissue, but it is not of general interest to the group, so I would like to send it to G.Arentieri directly. Greg, Could you try and contact me again? This time type your e-mail address just in case of another snafu. Bye, Hildy
I for one would be very interested in Hildy Crowley's opinions on long term tissue storage. Others ...?
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 e-mail: mcauliff-at-umdnj.edu ***************************************************************
The first thing I was wondering: Is the epitope extra or intra cellular? I guess the most success I've had is by manipulating the fixation. Trying the solvents vs. 2-4% paraformaldehyde or something like Zambonies or Bouins so the picric acid can go in fast and stablize the protien then the paraformaldehyde lightly crosslinks. Then if you need to open up the cell I use .1% Tween or to open epitopes a very short 1- 3min .01% Trypsin.
Bob Morphology Core
On Thu, 24 Jul 1997, kelly dowhower wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I have been transfecting mammalian cells with a dopamine receptor and } trying to detect, via fluorecence microscopy, its expression with a } polycolonal antibody that I made to one of this dopamine receptor's } intracellular loops. (The antibody detects the dopamine receptor epitope, } to which the antibody was made, on western blots, when expressed by E. coli } in the context of a fusion protein, so I know the antibody does recognize } the epitope against which it was made.) } } When I express the receptor in transfected HEK293 cells, fix the cells } by methanol:acetone, and do fluorescence microscopy, I get no detection of } receptor expression. I engineered a " FLAG tag" into the receptor and } stained for FLAG labeling;the FLAG epiope gave intense staining, so I know } the dopamine receptor is being expressed. } } Does anyone have any thoughts on what factors I might to alter in my } immunofluorescence assay in order to make the dopamine receptor antibody } recognize the dopamine receptor in the transfected cells? It seems as if } the receptor is folded in a conformation such that the epitope that the } antibody recognizes is buried / not exposed. Are there better/more } appropriate fixation methods? Does anyone have information on live } staining or microwave heating/steaming of the cells? } } Thanks for your input. } } Kelly Karpa } kjd136-at-psu.edu } } }
I have an undergraduate student working on the design of a homemade backscatter diffraction pattern for the SEM. We are trying to get patterns using a JEOL T300. We tried using an accelerating voltage of up to 30keV, turned up the spot size, turned down the gun bias, and we get a "bright" screen but no kikuchi lines. We tried different specimen tilts, specimen working distances, and screen to specimen distances, and the specimen is a good quality Si wafer, but nothing seems to produce a bright enough signal to generate the Kikuchi lines. Are we limited by the SEM we are using, or are we just missing something?
Thanks. Lucille Giannuzzi
************************************************************************* Lucille A. Giannuzzi, Ph.D. Assistant Professor
Dept. of Mechanical, Materials, and Aerospace Eng.
University of Central Florida phone (407) 823-5770 PO Box 162450 fax (407) 823-0208 4000 Central Florida Blvd. email lag-at-pegasus.cc.ucf.edu Orlando, FL 32816-2450 USA *************************************************************************
Zip drives are not yet discontinued...but they probably have a shorter market life-span than CD-R's (hard to predict the future though).
Optical drives are neat for archiving...but who has an optical drive on thier home/office system?? They are rather pricy.
CD-R's are the way to go I believe. Almost all computers these days have CD devices. The Disk and the drives are rather inexpensive. The storage life of a CD (not in vacuum) is over 60 years (and far longer in Vacuum storage). We store Tiff files and use HTML frontends so that almost ANYONE with ANY SYSTEM with ANY SOFTWARE can use them (OK...maybe not...but I have heard of no problems so far). They just make sence. We do not use a dedicated machine...But I would suggest to anyone that they at least use a dedicated h-drive for writing to (helps alot). I guess if a lab does not allow alot of access to archives then maybe an Optical drive would be better (less need for compatability).
On the subject of archiving medium going out of date:
At first thought one might think this is a horrible problem. We have about 200 old 8 inch floppies and no drive to read them. Further...I am not so sure how many of them are any good. They contain alot of years of data.
Yet, and I know this may not apply to everyone, what is the data worth?? I have found that the archiving Medium and devices seem to out last the Data. Any Data around here that are more than about 10 years old are almost worth-less. The machines used to collect them are long since replaced with better machines...we would have to recollect any archived data that old. Further, almost all older data worth note has been published and exists in hard copy some place.
Its neat to think that 100 years from now, someone some place will want to see some of my data, or a perfect BSE image I took. When that thought comes to mind I get a bit excited...then suddenly a new thought appears, "Yeah Right Christopher!! Keep Dreaming!". Think about the machines that will be in use 100 years from now.
I'll start with some notes on Wednesday's sessions since I neglected to summarize them last night. I was, er, a bit slow by the time I got back. :-P The session focus on Wednesday was "History and Art," although that was only loosely held to. Several of the papers dealt with artistic subjects but not art conservation per se.
Gary Laughlin spoke about "A Unique Metallurgical Process From the Early Bronze Age" in which he described the findings at, and significance of, a site excavated at the Kestel Mine in the Taurus Mountains of Turkey. The site dates from the third millenium BC. The examinations indicate that cassiterite ore was mined and refined at the site to yield a black magnetic oxide. This would have been more readily separated, due to its magnetism, than could be accomplished by other means.
Dr. McCrone gave an "Update on the Turin Shroud" in which he reviewed several letters he received from Father Rinaldi prior to the Father's death in 1993. In the letters, Father Rinaldi effectively acknowledged the proof that the Shroud was painted and actually dated from much later than the time of Christ's crucifiction, thus was not the true article.
(For those of you who may not know, Dr. McCrone concluded early on in the Shroud investigations, and from microscopic observations alone, that the shroud was a painting. He stood nearly alone in this view and was vilified for nearly two decades before Carbon dating ultimately confirmed his conclusions.)
John Delly gave one of his typical, amazing presentations, this one on "Hand-colored Microscopical Illustrations." In it John took his audience on a delightful stroll through 19th century microscopy publications illuminated with hand-colored illustrations. He demonstrated the evolution and later de-evolution of the quality of such illustrations, the variation that one can see from different illustrators of the same work, and the differences that are seen edition-to-edition of the same work. Of course, when he became interested in the subject, John felt compelled to master the art himself. Through his own study and practice he gained the insignt necessary to understand and explain the techniques and variations seen in these historical works. The illustrations are, indeed, beautiful and many of us are fortunate to own examples of these illustrations in early works on microscopy. Because of the vast number of color illustrations necessary to address his subject, it is unlikely that this presentation will ever be recorded fully in print. (How about a book, John?) Those of us fortunate enough to be in the room may be the only ones ever to enjoy this particular product of John's efforts. Thank you, John, once again.
Have you ever stood befor an audience wondering why on earth you found yourself presenting in the particular session where you were? Wayne Moorehead must have when he spoke on "A Tale of Two Danas; Influences in Mineralogy" but he soldiered on and did a fine job chronicaling the lives of two remarkable men. The mineralogists in the audience will need no introduction to the Danas but I'll just mention for the others that the elder Dana published his first edition of the System of Mineralogy in 1835 at the tender age of 24. It was the first such major scientific work of classification written in English and he and his son went on to publish or edit a vast array of classic works in Mineralogy. Together or individually, they edited the prestigious American Journal of Science continuously for an astounding 95 years, from 1840 to 1935. One of the most noteable achievements that Wayne mentioned, in my mind anyway, was when James Dana, in the introduction to a revised edition of his classic System, abruptly abandoned his entire earlier classification system as outdated!. Believing that system no longer consistant with emerging thought, he just as abruptly adopted and described a newer system which largely stays with us today. I find such honest self-appraisal and the ability to continue to move forward without missing stride quite refreshing.
Wednesday afternoon was occupied by two sessions which would be unusual at other meetings. Using video microscopy, Anna Teetsov of McCrone Associates demonstrated some micromanipulation techniques within the context of creating artistic works on microscope slides by arranging butterfly scales of various colors into micro-images. Anna and a few others continue to develop this art form which is particulary unique to the community of microscopists. One has to have a microscope and micro-related knowledge and skills in order to produce these beautiful little creations, then one has to have a microscope to view them as well. Kind of nice, don't you think? Something we can hold purely for the aesthetic pleasure and uniquely our own.
The afternoon was closed with a demonstration by Alan Shin on how one can construct a working replica of Leeuwenhoek's single lens microscopes. I did not attend this demonstration as I have on a previous occasion taken a longer version from Alan in which we got to actually construct our own microscopes. Comments from those who did attend and look through the instrument Alan made reflected surprise at how much one could see and pleasure at the experience of seeing an insturment of such historical significance actually fabricated.
Today's sessions were on Forensic Microscopy. Jose Almirall told us about "Developments in Glass Examination: Automated Microscopy Techniques and Composition Analysis." Jose's talk was very interesting and perhaps somewhat troubling to practicing forensic scientists as he told us of (among other things) a remarkable consistancy in the optical properties of some glasses, especially window glass manufactured by the "float" process. The new information for me was the time over which the products of these plants will remain indistinguishable under conventional forensic examination techniques. I am not aware of other time-dependant studies of glass properties but Jose showed data collected over at least 18 months, during which the product of a float glass plant was entirely uniform in refractive index. He did however, offer a remedy for this disturbing finding. He showed that glass samples which are indistinguishable by refractive index can often be distinguished by elemental analysis of Fe, Mg, Al, and Zr using ICP/AES. Now all the forensic people have to do is get themselves one of these and... ;-)
Wayne Moorehead gave another excellent paper on Thursday, this one on "An Introduction to Microscopical Feather Identification." Wayne told us that the flight and tail feathers of birds are not always useful for identification but that the down or contour (breast) feathers can be distinctive, at least down to the order of birds, occasionally to the family, but virtually never to the genus or species. Still, identification at this level may prove very useful in a forensic case. Wayne illustrated how appropriate preparations can be made, what features of the feather barbule to examine and how they can vary. He also showed and described the identifying characteristic of numerous feather types.
Thom Hopen gave an interesting talk on "Teaching Forensic Microscopy in Countries Formerly Known as the Soviet Union." Thom responded to a State Department request that he make numerous trips to various countries of the former Soviet Union. He has taught courses of fiber and paint comparison, explosives residue analysis, and basic microscopy. He found his students to be highly motivated and dedicated people, anxious for quality instruction in basic forensic microscopy techniques. Often they are at least adequately equiped though sometimes have little or no idea how to fully exploit the equipment they have. (Unfortunately, when it comes to microscopy, this is too often true here also! My comment, not Thom's.) One can only immagine the difficulty of teaching in a completely and literally foreign environment, working through a translator, and using instrumentation previously never seen. Often, Thom had to set the equipment in proper working order prior to beginning instruction. But apparently all has worked out for him and his students and several more trips are planned to continue the education.
Well folks, I think I'll stop there. Of course, there were many more fine presentations and, once again, I'll mention that my choice of topics covered here in no way reflects badly on the other papers. All of the presentations were excellent.
Tomorrow is given over to a tutorial workshop on the Dispersion Staining technique. It will be given at McCrone Research Institute by Dr. McCrone and will be attended by twenty-odd students, all that can reasonably be accomodated in such a hands-on session. For the rest of us, this afternoon marked the end of another educational, enlightening, and just plain fun Inter/Micro.
Special thanks, as usual, to Nancy Daerr who coordinates all arrangements for these meetings and who, as usual, did an exceptional job of taking care of us and making our stay wholly enjoyable.
To all of those interested in these meetings, please note: Next year marks the Golden Anniversary of Inter/Micro, the fiftieth anniversary. (Wow!) Plan on attending what promises to be an excellent meeting. Contact Nancy Daerr for further information, to be put on a mailing list, etc. She can be reached at McCrone Research Institute, 2820 S. Michigan Avenue, Chicago, IL, 60616 or simply as ndaerr-at-mcri.org. The phone numbers at McRI are 312-842-7100 (voice) and 312-842-1078 (fax).
It's been a pleasure being your ears at Inter/Micro 97. But tomorrow... Ahhh, Chicago! The architecture, the museums, the Art Institute! I feel like a nice walk! Happy trails to all, and to all, Good Night.
Steve Shaffer MicroDataware sshaffer-at-microdataware.com
Please post or pass this on as appropriate. Thank you.
Regards,
John Vetrano
----------
Electron Microscopy Positions in Materials Characterization
Pacific Northwest National Laboratory
The Structural Materials Research Group at Pacific Northwest National Laboratory is seeking applicants for a staff position and a post-doctoral appointment, both specializing in the characterization of materials using transmission electron microscopy (TEM). The Electron Microscopist staff position requires a minimum of a 2-year specialized degree with expertise and training in the operation of analytical TEM instruments as well as in the preparation of electron transparent materials for examination. This individual will conduct research on a wide variety of metallic, ceramic and composite materials in support of scientists and engineers in the Structural Materials Research Group. Current activities include both basic and applied research in the areas of deformation mechanisms in metals, interfacial segregation and precipitation, radiation effects in metals, ceramics and composites, and enviromental degradation. In addition, the position will help oversee the maintainance of the Group microscopy facilities which include three conventional TEMs, a field-emission-gun (FEG) TEM and a scanning TEM, SEMs and a scanning Auger microprobe.
A post-doctoral position is also available focussed on the high-resolution characterization of grain boundaries in metallic alloys by analytical TEM. Energy dispersive x-ray and electron energy loss spectroscopies will be used with the FEG-TEM to elucidate segregation and precipitation mechanisms in aluminum and stainless steel alloys. The initial post-doctoral position would be for one-year and a second-year renewal is possible. The position requires a PhD degree in material science, physics or a related discipline and experience in analytical TEM for quantitative compositional analysis.
Pacific Northwest National Laboratory (PNNL) is operated by Battelle Memorial Institute for the U.S. Department of Energy. PNNL is a multiprogram laboratory located at the Hanford site near the junction of the Columbia, Snake and Yakima rivers in SE Washington state.
For consideration, please submit a resume (including references) and publication list by Oct. 1, 1997 to: Dr. Stephen M. Bruemmer, Pacific Northwest National Laboratory, Battelle Boulevard, P.O. Box 999, Richland, WA 99352. Phone: (509) 376-0636; Fax: (509) 376-6308; e-mail: sm_bruemmer-at-pnl.gov.
Using different Apple Macintosh Workstations and one mobile Apple Powerbook 1400cs (which has an 8xCD-ROM drive), the CD-R Backup solution proved perfect for Software and Data Backup, and taking complete backup with the Powerbook still leaves me very mobile.
I use the Philips 2660 CD-Burner with Astarte Toast CD Software. This not only allows for different formats, but also for creation of CD-ROM image files on harddisk for easy preparation of a 650M partition, or smaller. If the Macintosh is setup well, and the driver software is properly installed (I use the FWB-CD-driver although not recommended by Astarte), no problems are expected to occur.
To deal with the fact, that some older machines do not read multi session CD-ROMs, I installed a 1G-Harddisk to collect the data to be burned on the CD on a 650M image file. As soon as it is full, I burn 2 CDs (one for regular use, one for backup), and throw the image file away. This proved itself useful, because at the time I burn the CD, some files are no longer needed and can be thrown away before burning.
A known fact with Floppy Disks is the time they keep the data until they begin to loose it (1-2 years? depending on brand?). This issue might be important and also brand-dependent in CDs as well, but the extent, the expected timerange and hence significane is unknown to me.
Wolf Schweitzer, MD Research Fellow, VIFM, Melbourne, Australia wschweitzer-at-access.ch
This listserver is a great help to me as an amature microsopist. I enjoy the information obtained by reading the daily "chat". I work 8 hours a day in the computer business. My suggestion is never trust any technology of today to last very long. The abiliity to store files for a long time is worthless if the technology on which it is stored is antiquated by as much as 3 - 5 years.
Zip drives are hot technology right now, but becauase of their limitations of size, at this point, they are being passed by with upcoming technology. File storage is good for the time being, but in less than a year look for the most modern and up to date file storage technology to change. Computers and their technology are always changing and becoming less expensive as the R&D is paid off and manufacture of newer ideas comes into being.
Do not hitch your wagon to anything as a forever thing. If silver grain photographs are s "thing of the past", whatever we use today will be also, very soon. CDRom drives and platters are fantastic, but technology to handle 1000 times the information storage in a smaller package via opti-digital technology is somewhere close around the corner.
I am not a proponent of either silver grain or digital file storage. Know that your will have to provide the latest thing for your business client there is and that will always change. Two years ago 4X CDRom drives were all the rage. Today you have to invest in a 16X or 20X drive that is less expensive than the 4X was. 4 gigbit hard drives retail for less than half the cost of a 540 megabit drive 5 years ago.
Sometimes it dosen't matter if the data will actually ever be accessed. It must just be available. Certain regulations (QA type stuff) can require data archiving for 30 years or more.
Any target the correct diameter will work. What I have done is clean-up the spent target assembly, then silver-epoxy a new (generic) disk to the target assembly face. Most EM suppliers sell disks. Material can also be purchased from suppliers like Alfa Aesar (Johnson Matthey) or Goodfellow. There is a supplier in Nevada (I think) who is reputed to be less expensive, but I don't have any more info available...Maybe someone else knows that name/address.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear all, I need some information. I have a Hummer VI-a sputter coater and I am having trouble finding where to get targets. EMCORP which sold the instrument seems to be out of business. Their phone numbers do not work. Any help will be appreciated.
TIA Greg Rudomen University microscopy Imaging Center S.U.N.Y. -at- Stony Brook
Greg wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Dear all, } I need some information. I have a Hummer VI-a sputter coater and I am } having trouble finding where to get targets. EMCORP which sold the } instrument seems to be out of business. Their phone numbers do not work. } Any help will be appreciated. } } TIA } Greg Rudomen } University microscopy Imaging Center } S.U.N.Y. -at- Stony Brook
Greg, We supply targets for all models of Hummer sputter coaters and almost any other type you might need. Please let me know what material and I will send you a quote. Thanks, JD Arnott Ladd Research
Greg wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Dear all, } I need some information. I have a Hummer VI-a sputter coater and I am } having trouble finding where to get targets. EMCORP which sold the } instrument seems to be out of business. Their phone numbers do not work. } Any help will be appreciated. } } TIA } Greg Rudomen } University microscopy Imaging Center } S.U.N.Y. -at- Stony Brook
Greg, We supply targets for all models of Hummer sputter coaters and almost any other type you might need. Please let me know what material and I will send you a quote. Thanks, JD Arnott
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear all, I need some information. I have a Hummer VI-a sputter coater and I am having trouble finding where to get targets. EMCORP which sold the instrument seems to be out of business. Their phone numbers do not work. Any help will be appreciated.
TIA Greg Rudomen University microscopy Imaging Center S.U.N.Y. -at- Stony Brook
Message-Id: {3.0.1.32.19970725104540.007717d4-at-mmserver.mm} X-Sender: opmills-at-mmserver.mm X-Mailer: Windows Eudora Pro Version 3.0.1 (32)
Hello,
I'm posting this query for a colleague here at MTU. Please contact him directly at qchorn-at-mtu.edu. Thanks.
Owen
++++++++++++++++++++++++
While characterizing the microstructure of a commercial ferrosilicon alloy, an interesting contrast event was observed within the silicon phase. The center of the silicon dendrites appears brighter than the edges when secondary electron imaging is used. This contrast is not observed using backscattered electron imaging. The following web site has images showing this contrast as well as other information about the alloy being examined:
http://www.mm.mtu.edu/~qchorn/siliconquestion.htm
Any insight as to what could be causing this contrast would be greatly appreciated.
Thanks,
Quinn C. Horn ++++++++++++++++++++++++++ Owen P. Mills Michigan Technological University Metallurgical & Materials Engineering Rm 512 MME Building Houghton, MI 49931 906-487-2002 906-487-2934 FAX opmills-at-mtu.edu
I agree with your comments that we sometimes try to save "all" of the data we generate, and much of it is of little future value. But having worked in the medical device industry for 20 years now, I would like to inject my two cents, just to provide another viewpoint.
In industry, some data (regardless of its age) is absolutely invaluable. An example would be data that is generated to support studies of the safety/efficacy of an implantable device. (Silicone breast implants are a good example). If 20 years from now, litigation arises with regard to one of our products, raw data may be required to lend credence to our studies. In fact, the FDA requires that we maintain certain data "forever", and some of this data is electronic. With this in mind, I have wrestled with this topic of data archiving and integrity for several years now. I try to ensure future compatibility of whatever software and hardware upgrades I make in our data collection equipment, but it's not always possible. Consequently, I have made it a point to also archive some of the older hardware and software, just in case.
Best Regards,
Bob ********************************** Bob Citron Chiron Vision Claremont, CA USA (909)399-1311 Bob_Citron-at-cc.chiron.com **********************************
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
On the subject of archiving:
Zip drives are not yet discontinued...but they probably have a shorter market life-span than CD-R's (hard to predict the future though).
Optical drives are neat for archiving...but who has an optical drive on thier home/office system?? They are rather pricy.
CD-R's are the way to go I believe. Almost all computers these days have CD devices. The Disk and the drives are rather inexpensive. The storage life of a CD (not in vacuum) is over 60 years (and far longer in Vacuum storage). We store Tiff files and use HTML frontends so that almost ANYONE with ANY SYSTEM with ANY SOFTWARE can use them (OK...maybe not...but I have heard of no problems so far). They just make sence. We do not use a dedicated machine...But I would suggest to anyone that they at least use a dedicated h-drive for writing to (helps alot). I guess if a lab does not allow alot of access to archives then maybe an Optical drive would be better (less need for compatability).
On the subject of archiving medium going out of date:
At first thought one might think this is a horrible problem. We have about 200 old 8 inch floppies and no drive to read them. Further...I am not so sure how many of them are any good. They contain alot of years of data.
Yet, and I know this may not apply to everyone, what is the data worth?? I have found that the archiving Medium and devices seem to out last the Data. Any Data around here that are more than about 10 years old are almost worth-less. The machines used to collect them are long since replaced with better machines...we would have to recollect any archived data that old. Further, almost all older data worth note has been published and exists in hard copy some place.
Its neat to think that 100 years from now, someone some place will want to see some of my data, or a perfect BSE image I took. When that thought comes to mind I get a bit excited...then suddenly a new thought appears, "Yeah Right Christopher!! Keep Dreaming!". Think about the machines that will be in use 100 years from now.
I just want to publicly thank Steve Shaffer for the interesting summaries of the talks at Inter/Micro. I hope I speak for all when I say that those of us who (for whatever reason, be it time, money, other committments) could not attend, really appreciated it. Hope you enjoyed the "off" time as well!
Best Regards,
Bob ************************** Bob Citron Chiron Vision Claremont, CA USA Bob_Citron-at-cc.chiron.com **************************
Has anyone thought of something useful to do with all the leftovers from dye sub printers? I have boxes of used ribbons and other things that seem too good just to toss out. Could they be used for something, or maybe recycled somehow?
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
Hi everyone, I have a couple of announcements about tickets to the Cleveland Indians game on Tues evening. One - If you have reseved tickets, I need to see the money!! Please send a check made out to 'MSA' for $20/ticket (cheap seats, but includes a picnic) to me here at:
Ferro Corp. 7500 E. Pleasant Valley Rd. Independence, OH 44131
Two - If you plan on canceling your reservations, I've had a number of requests for more tickets...So - I'm starting a waiting list. First come, first on the list. I have had a few cancelations, so far - so please give me a call at (216)641-8585 x6613. E-mail address is: vbauer-at-ferro.geis.com.
I know this isn't "Microscopy" but I don't have current e-mail addresses for everyone, so please bear with me.
One woman that used to work in our lab thought the ribbons would be good for Halloween--go to a party as the "Primary Colors"
Thomas Moninger moninger-at-emiris.iaf.uiowa.edu University of Iowa Central Microscopy Research Facility http://www.uiowa.edu/~cemrf Views expressed are mine. On Fri, 25 Jul 1997, Jon Krupp wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hi: } } Has anyone thought of something useful to do with all the leftovers from } dye sub printers? I have boxes of used ribbons and other things that seem } too good just to toss out. Could they be used for something, or maybe } recycled somehow? } } Jonathan Krupp } Microscopy and Imaging Lab } University of California } Santa Cruz, CA 95064 } (408) 459-2477 } FAX (408) 429-0146 } jmkrupp-at-cats.ucsc.edu } }
Subject: Time:12:45 PM OFFICE MEMO Philips 201 TEM Date:7/25/97
If anyone is interested in obtaining a Philips 201 (24 years old, on service contact continually) for a nominal cost, please contact me immediately. Ann E. Rushing Department of Biology Baylor University Waco, TX 76798 Ann_Rushing-at-baylor.edu (254) 755-2911
Bob Citron wrote: ================================================== I agree with your comments that we sometimes try to save "all" of the data we generate, and much of it is of little future value. But having worked in the medical device industry for 20 years now, I would like to inject my two cents, just to provide another viewpoint. ================================================== Actually, it is not just the medical device industry that has these concerns . The entire industry of analytical and testing laboratories have these concerns as well. You see, in our litigious society in America, there is no such thing as a "statute of limitations" for professional liability exposure (negligence as some would say). The "clock" starts ticking, not when the alleged error occurred, but when the alleged error is discovered. This means, in our case, we have to maintain complete records of all work done going back to the inception of our business in 1970 with all of the associated costs.
When these records are not maintained or are not kept available in retrievable form, then someone who is trying to make it seem like you have made some error years ago, really is in the driver's seat, that is, they can say whatever they want to say and you have nothing in your files that says otherwise. Worse yet, the people who did the work might not remember what really did go on, say twenty years ago, or can be either retired, no longer alive or just not anywhere to be found.
And what has to be kept is not only the original data from specific samples, but also the ancient records showing service history, instrument calibrations, and other test and measurement procedures that document that the instrumentation was operating the way it was claimed to have been operating.
Are there really instances where old archival information of this type is actually needed? You bet. Do laboratories or individual consultants get sued? You bet. And how do they fare without such supporting data and information? Not very well.
Disclaimer: From 9/76-3/97 served as shareholder and director of ILAC Ltd., Hamilton, Bermuda, an insurance company that insures analytical and testing laboratories for professional liability risk.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI Structure Probe, Inc. FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Dear All, The supplier of sputter coater targets in Nevada is Abe Dayani of Refining Systems Inc. P.O. Box 72466 Las Vegas, NV 89170 phone: 702-368-057 fax: 702-368-0933 He is a buyer of estate jewelry and will make discs of any precious metal or alloy for much closer to the list price of the metal than most suppliers. I believe that if you send him a target he will stick the new disc on it. Woody wrote: } Any target the correct diameter will work. What I have done is clean-up } the } spent target assembly, then silver-epoxy a new (generic) disk to the target } assembly face. Most EM suppliers sell disks. Material can also be } purchased } from suppliers like Alfa Aesar (Johnson Matthey) or Goodfellow. There is a } supplier in Nevada (I think) who is reputed to be less expensive, but I } don't } have any more info available...Maybe someone else knows that name/address. } } Woody White } Mcdermott Technology Inc. } } Alt: } woody.white-at-worldnet.att.net } http://www.geocities.com/capecanaveral/3722 } ______________________________ Reply Separator } _________________________________ } Subject: Hummer VI-a sputter coater } Author: greg-at-umic.sunysb.edu_at_internet at X400post } Date: 7/24/97 2:59 PM } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
For those of you who are not already familiar with it, I would like to introduce the Society for Luminescence Microscopy and Spectroscopy (SLMS). Members of this society are involved in cathodoluminescence (CL) and UV-excited fluorescence microscopy. Most of the members are involved with the earth sciences or the material sciences and ceramics. There are also some applications in forensics, archaeology, and other fields. The society has been in existence for about 10 years. There is a semi-annual newsletter, edited by Professor Kopp of the U. Tennessee. Dues are only $10.00 for full members and $5.00 for students. The instrumentation used includes the familiar SEMs and EMPAs and a significant number of the members are using relatively simple and inexpensive cold cathode based electron beam systems which are small enough in size that they can be mounted directly on the stage of a conventional monocular or binocular transmitted light microscope. The CL of the mineral samples can then be seen directly, in real time, with a minimum of sample preparation and the information revealed on impurity distributions, etc, is otherwise very difficult or impossible to obtain with other existing techniques.
The SLMS has a standards program which has dealt, so far, with the comparison of photographic results on a complexly zoned carbonate and with the comparison of the CL emission spectra from a Dy-doped zircon.
For more information on the SLMS, please visit our web site at http://zephyr.rice.edu/SLMS/SLMS.html This web site is maintained by Jinny Sisson at Rice University.
If you have any questions regarding the SLMS or the applications of CL, please feel free to contact me or one of the addresses on the web page.
Disclaimer: I am one of the many charter members of the SLMS and presently chairperson of the Standards Committee. I have a commercial interest in furnishing cold cathode based ebeam systems for CL investigations.
Donald J. Marshall
Donald J. Marshall Relion Industries P.O. Box 12 Bedford, MA 01730 Ph: 617-275-4695 FAX: 617-271-0252
To further comments from Robert H. Olley and Cynthia Bennett regarding plasma etching, I would like to share a little insight. Ion damage and "burn marks" are related to the type of plasma generation system. In the past several years there has been a significant amount of effort expended to reduce the ion energies present in the plasma. The greatest drive has been in the semiconductor industry which requires absolute cleaning or etching without any uncontrolled altering of the material being processed.
Two types of low energy plasma creation have been developed as a result, namely, inductively coupled and electron cyclotron resonance (ECR) which create ion energies of 10-15eV and 3-5eV, respectively. At these ion potentials, insufficient energy exists to either heat or alter the specimen. Organics are removed from the surface, or material is selectively removed from the specimen/substrate through the application of reactive gas species which are generated by the plasma. This process is purely chemical and does not rely on the forces of ion impingement to remove material.
To remove organics, oxygen is the ideal process gas which chemically converts hydrocarbons to CO, CO2 and H2O. To selectively etch materials, various process gases can be applied. This technology can be readily utilized without any concern over damage to the specimen. Robert Olley's application of the atomic oxygen generated in a radio frequency discharge began to touch on an appropriate application of plasma.
I hope that this clarifies some of the concerns over the use of plasmas.
Regards,
Paul
Paul E. Fischione E.A. Fischione Instruments, Inc. 9003 Corporate Circle Export, PA 15632 USA Phone (412)325-5444 FAX (412)325-5443 Web-site: www.fischione.com
Does anyone have a source for plastic slides/coverslips/dishes that can be used for tissue culture and won't autofluoresce? Some of the catalogues list products, but I haven't tried any of them and would like some feedback from the experts :)
Thanks to all who responded to my Hummer Sputter target question. Many of you have an adress for Anatech that is old. The new address is: Anatech LTD. 6621-F Electronic Dr. Springfield, Va 22151-4303 1-800-752-7629, Fax 703-941-8077
Greg Rudomen S.U.N.Y. at Stony Brook University Microscopy Imaging Center greg-at-umic.sunysb.edu
Greetings All, I'm curious about how often everyone aligns the beam on their 'scopes. I use a JEOL - JEM 100CX II. In the past, the alignment procedure was done every morning. Now, we only do it once a week. We don't seem to have any problems, and the photos are crisp. What are others doing?
Sharron G. Chism HT (ASCP) Electron Microscopy Lab Harris Methodist Hospital Fort Worth, Texas
} I'm curious about how often everyone aligns the beam on their 'scopes. } I use a JEOL - JEM 100CX II. In the past, the alignment procedure } was done every morning. Now, we only do it once a week. We don't seem } to have any problems, and the photos are crisp. What are others doing? }
I check alignment every morning. Hopefully any other problems will also show up.
Greg-at-unic.sunysb.edu University Microscopy Imkaging Center S.U.N.Y. Stony Brook
} ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com -----------------------------------------------------------------------. } } Greetings All, } I'm curious about how often everyone aligns the beam on their 'scopes. I use a JEOL - JEM 100CX II. In the past, the alignment procedure was done every morning. Now, we only do it once a week. We don't seem to have any problems, and the photos are crisp. What are others doing? } } Sharron G. Chism HT (ASCP) Electron Microscopy Lab Harris Methodist Hospital Fort Worth, Texas
If you have a multi-user environment with users of different experience levels, every user should perform the quick alignment procedures. When there is trouble and the machine can't be aligned with the simple procedures, then the manager or service engineer of the TEM should perform a complete one (which includes the mechanical alignment). In addition, the microscope should be left in a standard startup condition for the next user. (Aperture out, low mag, beam spread to uniformly light the screen, text on negative info screen erased, and the plate number and info changed to some default, e.g. 0001.)
I have recently used 10 micron frozen sections of muscle fibers for immunolabelling using an antibody produced in our lab that is specific for a protein on the myosin filaments. Visualization was achieved by a flourescent secondary antibody. The problem is that we are not able to obtain the resolution necessary with the flourescent antibody in order to access fiber to fiber variation in labelling within the sarcomere or between sarcomeres.
I am thinking of using a biotinylated secondary antibody such that a biotin/avidin horseradish peroxidase visualization system can be used. This would allow us to continue this study at the light microscope level. Eventually EM wil be utilized, but light microscopy will allow for broader asessment of this large muscle. Does anyone know of a dehydration and counterstaining protocol that can be used for final viewing of the muscle tissue?
As some of you may know, Alwyn Eades is leaving the position as Director of the Center for Microanalysis of Materials in the MRL for a faculty position at Lehigh University. We are very sorry to lose Alwyn, but he would like to affect a career change at this time and we wish him the best of luck.
We will be instituting a search for a new Director of the CMM. At this time, I am writing to you to inform you of this search and to ask that you notify any colleagues you feel might be qualified to apply. A description of the position is attached below. Please feel free to distribute it.
Alwyn has been an excellent Director of the CMM and to replace him we will need all the help we can get. I do hope you can help us.
Thank you in advance.
Howard Birnbaum
********************************************
PRINCIPAL RESEARCH SCIENTIST (DIRECTOR OF THE CENTER FOR MICROANALYSIS OF MATERIALS) FREDERICK SEITZ MATERIALS RESEARCH LABORATORY UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
The Frederick Seitz Materials Research Laboratory (MRL) at the University of Illinois at Urbana-Champaign is seeking a Director for its Center for Microanalysis of Materials (CMM). The CMM is a major analytic instrumentation center encompassing the techniques of electron microscopy (TEM, STEM, LEEM and SEM), microchemical and surface analysis (SIMS, Scanning Auger, XPS, UPS), ion beam methods (RBS, channeling, PIXE), probe microscopies (STM and AFM), and x-ray methods. It functions to support the research activities of faculty, Research Associates and students at the University of Illinois and at other universities, and for researchers at Federal Laboratories and in industry. The present staff of the CMM consists of thirteen professional analysts who are expert in the various analytic methods.
The Director of the CMM reports to the Director of the MRL and the CMM is supported by the MRL as one of a number of instrumentation centers. The CMM Director works with the Director of the MRL in planning the development of the CMM, in the development of new analytic methods, in the purchase of new and replacement instruments, and in making the CMM increasingly important in the research endeavor of the MRL. He/she is responsible for the management of the CMM staff, for providing scientific and technical expertise to the CMM, and with the CMM staff members, to the users of the CMM. The Director coordinates staff development and outreach activities to researchers at the University of Illinois and nationwide. She/he represents the MRL at appropriate national activities involving instrumentation facilities. The person selected for the position of Director is expected to carry out an appropriate research program (for which support is provided) and to encourage appropriate research activities on the part of the CMM staff.
Candidates should have a Ph.D. in Physics, Chemistry or Materials Science and have an established research record in an appropriate field involving the use of modern analytical methods. The candidates should have strong technical expertise in at least one of the areas of analysis covered by the CMM and should be capable of providing technical leadership in all of the analytic areas. He/she should have an established record of management of scientific personnel and the ability to develop and manage budgets.
The salary for this position will be commensurate with the experience of the candidate chosen and full benefits of the University of Illinois will apply. Please submit applications to: Howard Birnbaum, Director; c/o Ms Donna Jacobs; Frederick Seitz Materials Research Laboratory; University of Illinois at Urbana-Champaign; 104 South Goodwin Avenue; Urbana, IL 61801. The application material should include a resume', a statement of your views on the operation of a microanalysis facility within a large university research establishment such as the MRL, the names and contact information for five references who are familiar with aspects of your career important to your qualifications for the position. Applications received by November 1, 1997 will be fully considered but acceptance of applications and screening of applicants will continue until the position is filled.
The University of Illinois is an equal opportunity / affirmative action employer,
Howard K. Birnbaum (217) 333-1370 Materials Research Laboratory FAX (217) 244-2278 University of Illinois e mail: birnbaum-at-uimrl7.mrl.uiuc.edu Urbana, IL 61801
} I'm curious about how often everyone aligns the beam on their 'scopes. } I use a JEOL - JEM 100CX II. In the past, the alignment procedure } was done every morning. Now, we only do it once a week. We don't seem } to have any problems, and the photos are crisp. What are others doing? } Dear Sharron, We go through an extensive procedure daily. In addition for dif- fraction we also check the condenser aperture position and do further align- ment in selected area and diffraction modes. About once a month we check the mechanical alignment of the objective upper pole piece, and once a year we disassemble and clean the entire lens column and do a complete mechanical alignment. I don't think the normal user--doing imaging of thick biological specimens at ~10k mag--would notice the difference in pictures if we didn't do the alignment as often, but there are uses for which it is critical, and it tells us a lot about the machine. Since we do all the maintenance--no service contract for the HVEM--the information we get from doing frequent alignments is quite useful. Yours, Bill Tivol
} I'm curious about how often everyone aligns the beam on their 'scopes. } I use a JEOL - JEM 100CX II. In the past, the alignment procedure } was done every morning. Now, we only do it once a week. We don't seem } to have any problems, and the photos are crisp. What are others doing? } Dear Sharron, We go through an extensive procedure daily. In addition for dif- fraction we also check the condenser aperture position and do further align- ment in selected area and diffraction modes. About once a month we check the mechanical alignment of the objective upper pole piece, and once a year we disassemble and clean the entire lens column and do a complete mechanical alignment. I don't think the normal user--doing imaging of thick biological specimens at ~10k mag--would notice the difference in pictures if we didn't do the alignment as often, but there are uses for which it is critical, and it tells us a lot about the machine. Since we do all the maintenance--no service contract for the HVEM--the information we get from doing frequent alignments is quite useful. Yours, Bill Tivol
This is a multi-part message in MIME format. --------------842BF15519E83971302C20CF Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit
Greg wrote: } } Thanks to all who responded to my Hummer Sputter target question. Many } of you have an adress for Anatech that is old. } The new address is: } Anatech LTD. } 6621-F Electronic Dr. } Springfield, Va 22151-4303 } 1-800-752-7629, Fax 703-941-8077 } } Greg Rudomen } S.U.N.Y. at Stony Brook } University Microscopy Imaging Center } greg-at-umic.sunysb.edu
I generally do at least some cursory alignment procedures each time I sit at the microscope. Most of the time all is well, but sometimes the last person had it in a different mode and/or did not bring it back to a "standard state". However, like stretching before excercising, I also find that running through the alignment procedures gets me in the proper state of mind for doing microscopy (as well as allowing my eyes to get dark-adjusted). Therefore, I would run through it just for that purpose. Is that kind of a Zen thing? :-)
Cheers,
John Vetrano _______________________________________________________________________________
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Greetings All, I'm curious about how often everyone aligns the beam on their 'scopes. I use a JEOL - JEM 100CX II. In the past, the alignment procedure was done every morning. Now, we only do it once a week. We don't seem to have any problems, and the photos are crisp. What are others doing?
Sharron G. Chism HT (ASCP) Electron Microscopy Lab Harris Methodist Hospital Fort Worth, Texas
I started life as an EM service engineer in 1966, since then I think I ha= ve handled almost every type of commercial TEM and SEM, therefore I have bee= n able to take a good look at the subject of alignment.
How often you need to align a TEM depends very much on its stability, =
thermal and mechanical. If you leave the lenses switched on at a controlled temperature, about 2 deg centigrade below room temperature is ideal, there is no reason for constantly aligning the lenses. =
If the machine has mechanical lens alignment it is my experience that the=
more you move them the more they move, compromise! However you must do a=
quick check prior to use of 1. gun alignment - viewing the spot and halo= : 2. illumination - in relation to the screen centre: 3. condenser apertur= e - in relation to the screen when C2 is spread overfocus: 4. objective aperture - in relation to the diffraction spot.
Of course the alignments that you need to perform should relate to the levels that you expect to reach when using the instrument. Less than 10,000X then you can get away with a very quick check of saturation. =
Should your target be 500,000X then the most important alignment will be voltage alignment, as clearly resolution will be your goal. In this case=
forget using the instrument within 2 hours of switching on the high voltage; that is unless you have a gas filled HT tank. It takes this tim= e for the high voltage to stabilise due to heat gained in the tank being required to reach the same level as heat lost. Gas filled tanks seem to take about 45 minutes at 120kV!
Most people over align their instruments as if the feat of completing the=
alignment is part of their religion! =
The time when an alignment becomes critical is when the lens in question = or the high voltage become unstable. To mis align the lenses is a standard engineer trick to isolate a fault to a particular part of the instrument.= =
See my book "Maintaining & Monitoring the Transmission electron Microscop= e" published by the Royal Microscopical Society ISBN 0-19-856407-4. This bo= ok also outlines all the TEM alignment procedures, including deflection coil= s and stigmators as well as covering typical image defects due to alignment=
and instability problems.
In spite of what some manufacturers may claim the alignment of a TEM follows basic procedures which have not changed since the 1960s, how can they, they are just electron optics!
Greg wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Thanks to all who responded to my Hummer Sputter target question. Many } of you have an adress for Anatech that is old. } The new address is: } Anatech LTD. } 6621-F Electronic Dr. } Springfield, Va 22151-4303 } 1-800-752-7629, Fax 703-941-8077 } } Greg Rudomen } S.U.N.Y. at Stony Brook } University Microscopy Imaging Center } greg-at-umic.sunysb.edu
Web address of Anatech LTD is:
http://www.anatechltd.com
-- Henrik Kaker SEM-EDS Laboratory, Metal Ravne d.o.o. Koroska c.14, 2390 Ravne, Slovenia Tel: +386-602-21-131, Fax: +386-602-20-436 SEM-EDS Laboratory Web Site http://www2.arnes.si/guest/sgszmera1/index.html Microscopy Vendors Database http://www.kaker.com/mvd/vendors.html Kaker.Com http://www.kaker.com
In reference to Cynthia Bennett's and Robert H. Olley's information request, there are in fact commercially available multi-process radio frequency etching systems at reasonable costs. For a full range of etch capability, it is necessary to have multi-gas inputs for user determined ratios of more than one species and the versatility to select more than one etching gas. Various gases can be used for selective etch of specific materials as has been done in the semiconductor industry since the 1960's. There are a couple reference texts available with specific designs of etching systems including parallel plate, ECR, inductivly coupled plasma and microwave technology. "Thin Film Processes" by Vossen, version I (1978) and II is an excellent reference to these processes. Also "Glow Discharge Processes" by Chapman (1980) covers these techniques in detail. As indicated in other responses, oxygen is most common for etching or removing hydrocarbons, organics and polymers such as photoresist. The O2 gas provides a chemical etch and in some cases it is desired to speed up the process by adding a heavier, non-reactive atom such as argon. This will cause a physical etch as well as a reactive chemical etch process.
To avoid "advertising" on this network, please refer to the South Bay Technology home page for additional information on one commercial versatile RF plasma etching system or contact me direct for details on the unit.
Regards, Steve
Steve Collins scollins-at-southbaytech.com Ph: 703-486-7999 (east coast USA) 714-492-2600 (west coast USA) 800-728-2233 (Toll Free) Fax: 714-492-1499 Web Page: http://www.southbaytech.com
At 10:10 AM 7/28/97 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Column alignment of TEMs is mostly for convenience and does not much affect resolution.
Filament traverse and tilt are so a maximal amount of beam enters the condenser system
Condenser alignment is so the illumination stays on the screen when condenser lens focal length is changed. The condenser/gun system is aligned with the axis of the objective so the illumination stays on the screen as objective focal length is changed.
Imaging lenses have their axes put on the line joining the centre of the objective with the centre of the screen so the image stays central and in view when magnification is altered.
The objective aperture MUST be centred on the axis of the objective. We use the zero order spot in the diffration pattern to define the axis.
For best resolution the entire illumination system should be tilted so its axis coincides with either the current or voltage centre of the objective. Voltage centre has an advantage as it aligns with the mean axes of all the imaging lenses, not just the objective. But there is not much in it. So when you are shooting for really good resolution, centre the objective aperture, correct astigmatism using background phase speckle on your specimen, use high voltage modulation to test illumination tilt and adjust for minimal image wobble, and shoot.
Maybe once a week I check condenser aperture alignment, every operator must check objective aperture alignment, gun alignment is checked after filament change of voltage change. The rest doesn't matter. We have not done a full alignment on our Hitachi H-7000 in 7 years. Mel Dickson President, Australian Society for Electron Microscopy Director, Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
Received: from mx05.erols.com (mx05.erols.com [205.252.116.249]) by mail0.erols.com (8.8.5/8.7.3/970701.001epv) with ESMTP id WAA20834 for {stevesbt-at-mail0.erols.com} ; Mon, 28 Jul 1997 22:01:06 -0400 (EDT) Received: from asklepios.med.vu.nl (asklepios.med.vu.nl [130.37.208.3]) by mx05.erols.com (8.8.5/8.8.5/MX-mnd) with SMTP id VAA17038 for {stevesbt-at-erols.com} ; Mon, 28 Jul 1997 21:55:21 -0400 Received: by asklepios.med.vu.nl with VINES-ISMTP; Tue, 29 Jul 97 3:52:51 +0200
Hello All:
In reference to Cynthia Bennett's and Robert H. Olley's information request, there are in fact commercially available multi-process radio frequency etching systems at reasonable costs. For a full range of etch capability, it is necessary to have multi-gas inputs for user determined ratios of more than one species and the versatility to select more than one etching gas. Various gases can be used for selective etch of specific materials as has been done in the semiconductor industry since the 1960's. There are a couple reference texts available with specific designs of etching systems including parallel plate, ECR, inductivly coupled plasma and microwave technology. "Thin Film Processes" by Vossen, version I (1978) and II is an excellent reference to these processes. Also "Glow Discharge Processes" by Chapman (1980) covers these techniques in detail. As indicated in other responses, oxygen is most common for etching or removing hydrocarbons, organics and polymers such as photoresist. The O2 gas provides a chemical etch and in some cases it is desired to speed up the process by adding a heavier, non-reactive atom such as argon. This will cause a physical etch as well as a reactive chemical etch process.
To avoid "advertising" on this network, please refer to the South Bay Technology home page for additional information on one commercial versatile RF plasma etching system or contact me direct for details on the unit.
Regards, Steve
Steve Collins scollins-at-southbaytech.com Ph: 703-486-7999 (east coast USA) 714-492-2600 (west coast USA) 800-728-2233 (Toll Free) Fax: 714-492-1499 Web Page: http://www.southbaytech.com
I also have a 100CX. I align the microscope everytime I sit down to it. If it is well aligned already the whole time takes just a few minutes out of my day. It's just a good habit to get into particularly if you have a multiuser environment.
And please pardon the bandwidth if anybody feels it's inappropriate. = This is the easiest way to widely disperse the message.
I have a substantial supply of Epon 812 (the real thing) that I find I = must part with. If anyone is interested in retro-embedding their = specimens, please contact me privately.
If you have access to a microscope with DIC (differential interference contrast) You can visualize the muscle very nicely and see the peroxidase product. You can also counterstain with hematoxylin in get more reference points.
Bob Morphology Core
On Mon, 28 Jul 1997, Lisa Brown wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I have recently used 10 micron frozen sections of muscle fibers for } immunolabelling using an antibody produced in our lab that is specific for } a protein on the myosin filaments. Visualization was achieved by a } flourescent secondary antibody. The problem is that we are not able to } obtain the resolution necessary with the flourescent antibody in order to } access fiber to fiber variation in labelling within the sarcomere or } between sarcomeres. } } I am thinking of using a biotinylated secondary antibody such that a } biotin/avidin horseradish peroxidase visualization system can be used. This } would allow us to continue this study at the light microscope level. } Eventually EM wil be utilized, but light microscopy will allow for broader } asessment of this large muscle. Does anyone know of a dehydration and } counterstaining protocol that can be used for final viewing of the muscle } tissue? } } }
Hi folks We are trying to find the most economic source for FUji Pictrography supplies, I would welcome info on where current users buy their materials and how much they pay for the various components. I will compile the results and send back to anyone interested. If vendors wish to add to this cost survey please contact me directly Tx
-- Simon C. Watkins Ph.D. Associate Professor Director CBI University of Pittsburgh Pittsburgh PA 15261 tel:412-648-3051 Fax:412-648-2004 URL:http://sbic6.sbic.pitt.edu
We are in a tight spot. We have collected data with a monoclonal antibody (Snap-25, reactive with fusion proteins from COS cells - the immunogen was crude synaptic immunoprecipitate from humans) which was raised in mouse. We used rats for data collection. Our secondary was FITC-conjugated AFfiniPure Goat Anti-mouse IgG. Now we find that we cannot obtain a peptide for control purposes. And, if we omit the primary, and use only the above secondary, we get a definite immuno response: It looks exactly as though the primary had been applied. The manufacturer of the FITC states that the anti-mouse antibody may cross-react with other species. Now What? We cannot purchase polyclonal Snap-25. Does anyone have any ideas? Did we make a huge mistake in not testing for control situations at the outset? We are relatively new at this game - do others get down these blind alleys, singing and dancing all the way until the lights go out? Bye, Hildy
Thanks to all who responded to my question on the frequency of beam alignment. It was interesting to hear from all of you. My favorite response included the advice of "If it ain't broke, don't fix it!". The majority of those that answered do a quick alignment every day ... mostly because of so many pairs of hands that fiddle with the 'scope during the day. Since I am the only tech that works with this 'scope, and always return it to "square one", and only have two pathologists that operate it ... once a week alignment seems to be all it needs. Our normal operation takes us up to 14k or 20k mag and seldom higher than 60k - 80k. (The specimens are about 70nm in thickness.) Still others have said that they, too, have a CX100 and only align the beam once a week. The information has been very helpful. Thanks, again for your info!
Sharron G. Chism HT (ASCP) Electron Microscopy Lab Harris Methodist Hospital Fort Worth, Texas
Our Local Arrangements Committee is chewing over a question that on which we have differing opinions. The question is " What is the true function of the Sunday social and how does this function relate to the venue?" We have come up with a couple of options.
Option 1: Is this function serving to rekindle acquaintances between colleagues.
Option 2: Is this function serving to not only rekindle acquaintances between colleagues but also to set the tone for the coming meeting, and show case the hosting city.
For those list members that attend the meetings, and any others that would like to comment all your input will be welcome. If you want to reply directly to me, I will be delighted to post a summary after comments have been received.
Bob Kayton, Ph.D. Histology/E.M. Core Director C.R.O.E.T. Oregon Health Sciences University Portland, OR 97201 W-503-494-2504 Fax-503-494-6831 H-503-590-7801
Jackson Immunoresearch carries secondary antibodies that are pre-absorbed against several different animal seras including rat. We routinely do double label ICC with mouse and rat monoclonals and purchase our secodary antibodies from them. See Braisted et al. Development 120:2409-2419 (1994). There is no cross reactivity seen with the mouse secondary to the rat immunoglobins. These preabsorbed antibodies should work well on your mouse tissue. Other companies carry pre-absorbed secondaries also. We have been using Jackson for years and have been very satisfied with them. Linda Barthel Research Associate II Department of Anatomy and Cell Biology University of Michigan lab (313) 764-7476 fax (313) 763-1166 barthel-at-umich.edu
On Tue, 29 Jul 1997, HILDEGARD CROWLEY wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Dear Immuno LM Friends, } } We are in a tight spot. We have collected data with a monoclonal } antibody (Snap-25, reactive with fusion proteins from COS cells - the } immunogen was crude synaptic immunoprecipitate from humans) which was } raised in mouse. We used rats for data } collection. Our secondary was FITC-conjugated AFfiniPure Goat Anti-mouse } IgG. Now we find that we cannot obtain a peptide for control purposes. } And, if we omit the primary, and use only the above secondary, we get a } definite immuno response: It looks exactly as though the primary had been } applied. The manufacturer of the FITC } states that the anti-mouse antibody may cross-react with other species. } Now What? } We cannot purchase polyclonal Snap-25. Does anyone have any ideas? Did } we make a huge mistake in not testing for control situations at the outset? } We are relatively new at this game - do others get down these blind } alleys, singing and dancing all the way until the lights go out? } Bye, } Hildy }
In my opinion concerning 100C(X) alignment, don't make knots to your neurons! 100 cx is a very complete but simple microscope to use.Only 6 lenses and 4 alignments coils including condenser and objective stigmators. All I'm describing is for routine work, for High resolution job it's an other problem. The main mechanical alignment is intermediate and projector lens and must be perform once or two times a year. alignment of condenser aperture is OK until people change the aperture. Gun tilt and shift are generally OK until you change the filament. The only thing to check is Spot size 1 and center with Gun shift then spot size 3 and center with Trans and repeat (once or two time) until the position of beam is the same between spot 1 and 3. If you have turn a lot the gun shift, you can check the maximunm of brightness or the image filament with gun tilt. If the precedent user has made dark field you can check the voltage center with HV wobbler and tilt knob. That's all, no more than a couple of minutes. Of course centering of objective aperture and setting of objective stig must be perform by user but it's not alignment it's just like to focus the image to obtain the best you can have. Hope that helps. ========================================================== Jacky Larnould mailto:larnould-at-mnet.fr voice:33 (0)4 67 72 28 26 fax :33 (0)4 67 79 54 90
I am looking for information about the ISO 9001 standard with regard to the operation of FT-IR microscopy and SEM systems for materials analysis, and would very much appreciate communicating with list members who have practical experience in this area.
Hi all: EMSL has a Philips 400 TEM/STEM to give away to any qualified non-profit. The instrument is in good condition and is located in Greensboro, NC. Recipient is responsible for packing and moving the instrument. Interested parties should contact Ron Mahoney at EMSL 910-297-1487 for more information. Cheers, Julian
Julian P.S. Smith III Biology Winthrop University Rock Hill, SC 29733 803-323-2111 x227 (vox) 803-323-2246 (fax)
} Date: Tue, 29 Jul 1997 11:02:50 -0600 (MDT) } From: HILDEGARD CROWLEY {hcrowley-at-du.edu} } To: postmessage {Microscopy-at-sparc5.microscopy.com} } Subject: LM:Mess-Polyclonal-mono-control } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Dear Immuno LM Friends, } } We are in a tight spot. We have collected data with a monoclonal } antibody (Snap-25, reactive with fusion proteins from COS cells - the } immunogen was crude synaptic immunoprecipitate from humans) which was } raised in mouse. We used rats for data } collection. Our secondary was FITC-conjugated AFfiniPure Goat Anti-mouse } IgG. Now we find that we cannot obtain a peptide for control purposes. } And, if we omit the primary, and use only the above secondary, we get a } definite immuno response: It looks exactly as though the primary had been } applied. The manufacturer of the FITC } states that the anti-mouse antibody may cross-react with other species. } Now What? } We cannot purchase polyclonal Snap-25. Does anyone have any ideas? Did } we make a huge mistake in not testing for control situations at the outset? } YES
We are relatively new at this game - do others get down these blind } alleys, singing and dancing all the way until the lights go out?
SUGGEST YOU TALK TO AN IMMUNOLOGIST TO HELP DESIGN YOUR IMMUNOEXPERIMENTS. THERE ARE LOTS OF REACTIVITIES THAT EVEN EXEPRIENCED MICROSCOPISTS DON'T KNOW ABOUT WHICH ARE COMMON KNOWLEDGE AMONG IMMUNOLOGY FOLKS. FORTUNATELY, I MARRIED AN IMMUNOPATHOLOGIST WITH A PH D IN IMMUNOLOGY--HE SOLVES MY REACTIVITY PROBLEMS.
} Bye, } Hildy } Rats and mice are very similar. I'm not surprised you got cross reactivity. You should always do negative controls-- in every experiment--one of which is your secondary without the primary. But this is not enough; you should have a primary control (one that is the same type as your experimental), either a preimmune serum if polyclonal or a non-reactive mono (proven non-reactive) that is the same species and same isotype.
You can buy a rat antimouse-FITC which probably won't react with rat tissue (ours didn't). Also, you can try absorbing your secondary with normal mouse serum, but you will probably lose a lot of your specific reactivity too.
Good luck, 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
Anyone have any suggestions for the question below. This is out of my area...
Nestor
} Date: Tue, 29 Jul 1997 10:48:01 -0500 } To: Zaluzec-at-sparc5.microscopy.com } From: lmtuhela-at-cc.owu.edu () } Subject: Ask-A-Microscopist } } Below is the result of your feedback form. It was submitted by } (lmtuhela-at-cc.owu.edu) on Tuesday, July 29, 1997 at 10:48:00 } --------------------------------------------------------------------------- } } Email: lmtuhela-at-cc.owu.edu } Name: Laura Tuhela-Reuning } } School: Ohio Wesleyan University } } State: OH } } Zip: 43015 } } Question: A faculty member is interested in using our SEM to observe the } giant chromosome in Drosophila for an upcoming genetics class. Are there } any suggestions for preparing the samples? We have a cryo system } available as well as variable pressure but do not have a critical point } dryer. Thank you! } } --------------------------------------------------------------------------- }
We do a lot of immunocytochem on Rat tissue and Rat -derived cultured cells and have (almost) always had good reults using anti-mouse secondaries from Jackson Immunoresearch (I've no stake in this company). They have antibodies raised in Donkey that are already preadsorbed against a number of other species, including Rat. Generally they give very low non-specific binding on our specimens. Negative controls are very important. We use the "no primary" and "pre-immune (non-immune)" controls often, but prefer the "irrelevent primary" control where possible.
Greg Martin Dept. of Cell Biology and Anatomy Johns Hopkins School of Medicine
On Tue, 29 Jul 1997, HILDEGARD CROWLEY wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Dear Immuno LM Friends, } } We are in a tight spot. We have collected data with a monoclonal } antibody (Snap-25, reactive with fusion proteins from COS cells - the } immunogen was crude synaptic immunoprecipitate from humans) which was } raised in mouse. We used rats for data } collection. Our secondary was FITC-conjugated AFfiniPure Goat Anti-mouse } IgG. Now we find that we cannot obtain a peptide for control purposes. } And, if we omit the primary, and use only the above secondary, we get a } definite immuno response: It looks exactly as though the primary had been } applied. The manufacturer of the FITC } states that the anti-mouse antibody may cross-react with other species. } Now What? } We cannot purchase polyclonal Snap-25. Does anyone have any ideas? Did } we make a huge mistake in not testing for control situations at the outset? } We are relatively new at this game - do others get down these blind } alleys, singing and dancing all the way until the lights go out? } Bye, } Hildy }
Michigan State University has an electron microscope available it is a Philips model EM201, purchased in 1972. Anyone interested contact Roger Cargill (517)355-0364 or cargill-at-pilot.msu.edu. thank you
A collegue has been offered a free Zeiss 25. It would be shared between the biology group and the materials group and the cost to them would only be shared maintenance. He is interested in its use as a materials science TEM.
I'm looking for any information about this microscope and comments from those familiar with it. There was no information on any TEMs at Zeiss' web sites. Are they still producing them? What about the Omega filter?
Michael Cinibulk UES Inc. at Wright Laboratory Wright-Patterson Air Force Base, Ohio cinibumk-at-ml.wpafb.af.mil
I need to purchase 300 mesh copper and 1000micron-slotted or hole copper grids in the 2.3 mm size. Does any vendor in the US supply these? I know that Agar does in the UK but I thought it might be faster to purchase them on this side of the pond. Can anybody out there help me? Thanks so much.
Cheers, Peggy Bisher.
NEC Research Institute 4 Independence Way Princeton, NJ 08540. Tel.: (609) 951-2629 Fax: (609) 951-2496 e-mail: peggy-at-research.nj.nec.com
(1) there is no current manufacturer of 35mm unperforated orthochromatic film;
(2) we have been offered a supply of some old stock at reasonable price, but this will not last for all that long;
(3) it would be nice if we could pressurize some manufacturer into re-doing the stuff. The Agfa Scientia appears to be the best;
(4) a gentleman from Kodak in New York sent me a roll of TX100 to try in our optical microscope / SEM. This material allows a large range of greyscales, and it works! It did particularly well for micrographs between crossed polars, where objects type A are only just off extinction and in the same field, objects type B were displaying full birefringence. And the quality was good - "brilliant grey", if such a colour exists.
+------------------------------------------------------------------------+ | 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 | +------------------------------------------------------------------------+
} A collegue has been offered a free Zeiss 25. It would be shared between the } biology group and the materials group and the cost to them would only be } shared maintenance. He is interested in its use as a materials science TEM. } } I'm looking for any information about this microscope and comments from those } familiar with it. There was no information on any TEMs at Zeiss' web sites. } Are they still producing them? What about the Omega filter?
For Zeiss, see now: http://www.leo-em.co.uk/ or http://www.mwrn.com/leo/leocont.htm
Yves MANIETTE Universitat de Barcelona Serveis Cientifico Tecnics Unitat ESCA TEM Carrer Lluis Sole i Sabaris, 1-3 E-08028 BARCELONA ESPANYA
Our Lab is ISO certified and I run the (materials) SEM Lab. Since ISO seems to be oriented toward a manufacturing environment where repetitive steps are involved, I found it a "pain in the butt" for a research/failure analysis oriented SEM facility. Procedures is the "magic word" (translate that to lots of paper work). I have procedures for operating the SEM, EDS, and WDS and also for periodic calibration checks. These procedures should reflect what is necessary to operate the equipment and generate "good" data, but at the same time, be as general as possible. This is so that under unusual circumstances your analysis strategies and conditions are not too limited. Good records keeping and adherence to the procedures are closely scrutinized. Use of ASTM methods and NIST certified standards will "lubricate" the ISO approval procedure.
Good Luck! Woody White Mcdermott Technologies, Inc
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am looking for information about the ISO 9001 standard with regard to the operation of FT-IR microscopy and SEM systems for materials analysis, and would very much appreciate communicating with list members who have practical experience in this area.
Is anyone familiar with stains and protocols for tissues embedded in Spurr's low viscosity resin? I have been using toluidine blue on one microns and need to achieve a greater differentiation among collagen fibers and keratocytes within the stroma of the human cornea.
} ---------------------------------------------------------------------------- } } Welcome to } } Microscopy-at-MSA.Microscopy.Com } ----------------------------------------------------- } ---------------------- } } Hosted and Sponsored by The Microscopy Society of America (MSA) } } as a Free Service to the World-Wide Microscopy & Microanalysis } Community } --------- } ------------------------------------------------------------------ } Your subscription request to the Listserver/MailReflector has been } received. } Please reply to this message to confirm that your address has been } successfully recorded. When your reply is received your address will } be } added to the Mailing List. } } ********************************** } IMPORTANT } please send this reply to } LISTSERVER-at-MSA.Microscopy.Com } ------------------------------------------------ } DONOT send it to Microscopy-at-MSA.Microscopy.Com } otherwise everyone on the server will } see your message and this creates unnecessary } and annoying traffic!!!!!!!!!!!! } ************************************* } } The purpose of this reply is to insure that no typographical errors } have } occured during your subscription to the system and to minimize the } transmission of Email to incorrect addresses which sometimes result in } } messages being bounced/reflected to a large list of receipents tying } up the } system (and your mail account!). } } After sending in your reply all discussion/comments/questions which } you } would like to be distributed to the list should be done by sending the } text } as a simple Email message } } To: Microscopy-at-MSA.Microscopy.Com } } All subscribers to the list (including yourself!) will automatically } receive } copies of the message you post. } } As a courtesy to the readers of this list please indicate in the } subject } line of your message a reasonable descriptive title of your } comment/inquiry. } Also preface your description by the conventional abbreviation of the } type } of microscopy you are interested such as LM, IRM, XRM, TEM, SEM, AFM, } STM, } uProbe...... For example, if you are interested in optical microscopy } and } have a question about staining then a Title/Subject line for your } message } might be } } Subject: LM - Need help on Staining Cells } } or if you are interested in TEM analysis of dislocations then } } Subject: TEM - Dislocation Loop Analysis } and so forth. } } Address all problems, questions, subscriptions, confirmation replies, } and } unsubscriptions to: } } Listserver-at-MSA.Microscopy.Com } } Thanks for your Cooperation: } } Nestor J. Zaluzec } Email:Zaluzec-at-MSA.Microscopy.Com } Tel: 630-739-1160 } } ================= } ========================================================= } } --------------------------------------------------------- } ------------------ } } General Ground Rules on using the MSA Microscopy } Listserver/Mailreflector } } ------------------------ } --------------------------------------------------- } Before continuing you should understand your responsibilities as a } Microscopy Listserver/Mailreflector System user. } } Specifically they are: } } * 1.) Actively encourage and promote the free exchange and } discussion of } information, ideas and opinions, except when the content would } compromise the national security of the United States (or any } other } country for that matter) ; violate proprietary rights, personal } privacy, or applicable state/federal/local laws and regulations } affecting telecommunications; or constitute a crime or libel. } } * 2.) Use your REAL NAME and fully disclose any personal, } financial, or } commercial interest when evaluating any specific product or } service, or } contribution. } } * 3.) Do not use this system for delivery of personal mail, } messages or } items of a similiar nature (i.e no posting of resume's etc....) } If you } are not sure then it probably does not belong here! If you would } like } an opinion then Email the message to me and I will review and } comment } as appropriate. (Zaluzec-at-aaem.amc.anl.gov). } } * 4.) Adhere to these rules and notify the } Listserver-at-aaem.amc.anl.gov } immediately when you discover any violations. } } ======================================================= } Additional Information } ======================================================= } } Anonymous FTP access to files in the Microscopy and Microanalysis } Software } Library (MMSLib), and the Listserver Archives. } } FTP SITE ADDRESS : ftp.msa.microscopy.com } UserName : Anonymous } Password : Your Email Address } WWW SITE ADDRESS : http://www.msa.microscopy.com } ======================================================= } Now for some Legal stuff } ======================================================= } } Every user explicitly acknowledges that all information obtained from } this } system is provided "as is" without warranty of any kind, either } expressed or } implied, including, but not limited to the implied warranties of } merchantability and fitness for a particular purpose and that the } entire } risk of acting on information obtained from this system, including the } } entire costs of all necessary remedies, is with those who choose to } act on } such information and not with the operators of this system or the } suppliers } of the information. } } Email: Nestor J. Zaluzec } } ======================== } ================================================= } To: NewSub-at-MSA.Microscopy.Com } From: "Nestor J. Zaluzec" {zaluzec-at-microscopy.com} } Subject: FAQ File - Microscopy ListServer } Cc: } Bcc: } X-Attachments: } } -------------- } ------------------------------------------------------------- } Help/FAQ File } THERE IS ALOT OF IMPORTANT INFORMATION HERE. } MAKE SURE YOU READ IT ALL.... } ----------------------------------------------------- } ----------------------- } ************************************** } FAQ's File } Microscopy/Microanalysis Listserver: } ************************************** } Last Updated April 15, 1997 } ************************************** } } --------------------------------------------------------- } ------------------ } What is the purpose of this Listserver? } --------------------------------------- } ------------------------------------ } } The purpose of this system is to give the scientific community a } centralized } Internet (Email) address to which questions/comments/answers in the } various } fields of Microscopy & Microanalysis can be rapidly distributed to a } list of } interested individuals and organizations. } } ----------------------------------------- } ---------------------------------- } What type of Microscopy/Microanalysis does this List address? } ------------------------------------------------------------- } -------------- } } For the purposes of this discussion forum, Microscopy & Microanalysis } should } be considered to include all techniques which employ a probe such as: } photons (including xrays), electrons, ions, mechanical and/or } electromagnetic radiation to form a representation or characterization } of } the morphology, crystallography, elemental, chemical or electronic } structure } of any material in either physical and/or life sciences applications. } } Some of the more common techniques which are associated with these } fields } include the following: optical microscopy, xray microscopy, scanning } electron microscopy, transmission electron microscopy, atomic force } microscopy, scanning tunneling microscopy, scanning ion microscopy, } analytical electron microscopy, electron microprobe, xray energy } dispersive } spectroscopy, electron energy loss spectroscopy, electron diffraction, } } convergent beam diffraction, high resolution electron microscopy, high } } voltage electron microscopy, etc. (the list continues...ad infinitum.. } ) } } - } -------------------------------------------------------------------------- } } How do I subscribe? } ------------------- } -------------------------------------------------------- } } Send an Email Message to "Listserver-at-MSA.Microscopy.Com" in the body } of your } message include the lines } } Subscribe Microscopy "USERID-at-EMailAddress" Optional: Real Name } Optional: Real (Surface Mail) Address } Optional: Microscopy/Microanalysis Interests } } Replace the "USERID-at-EMailAddress" with your real Email address. You } may also } append your Real Name, and Address etc.. to the end of the message as } a } courtesy. I'd be interested to know who you are and where you're from, } and } what you do. } } ------------ } --------------------------------------------------------------- } How do I unsubscribe? } --------------------- } ------------------------------------------------------ } } Send an Email Message to "Listserver-at-MSA.Microscopy.Com" in the body } of your } message include the line } } Unsubscribe Microscopy "USERID-at-EMailAddress" } } Replace the "USERID-at-EMailAddress" with your real Email address. } } IMPORTANT NOTE#1. To unsubscribe you must supply the original address } which } you used when you first submitted your subscription. Over the last few } years } I have found that many users have their mail forwarded to new } addresses, or } use an alias. When they then try to unsubscribe, they forget where } there } mail is being sent to (or forwarded from). This causes me lots of } headaches. } So just remember in order to unsubscribe you from the list, the } listserver } must know the your original subscription address to remove it! } } Please be considerate. I am not a mindreader, and don't really enjoy } playing } detective over the Internet. I have too many other things to do. } } IMPORTANT NOTE #2: Do not send subscribe/unsubscribe message to } "Microscopy-at-MSA.Microscopy.Com". When you do this ALL members of the } listserver receive your message. All administrative issues should be } sent to } "Listserver-at-MSA.Microscopy.Com" and not to the general mailing list. } } -------------------------------------------------------------------- } ------- } Are There Any Special Settings on my Email Program? } --------------------------------------------------- } ------------------------ } } Yes. } } 1.) Please insure that your correct Email address is listed in the } FROM: } record. You may need to enter this in the Preferences option of your } Email } program. Contact your system administrator if your not sure what to do } here. } } 2.) DONOT set your Email Program to automatically reply to all } messages, or } to request a return receipt. If either of these are done, you run the } risk } of creating an Email loop within the system. This may result in a } message } bouncing through the system for several days, filling up everyone's } mail } box! } } 3.) Generally it is best to terminate each line of your message at } less than } 80 characters. This avoids the occassional problem of messages getting } } chopped up. Some mail routers, limit the number of characters per } line. If } you exceed 80 characters it is sometimes possible that your message } will } become randomly truncated. If your not sure, then simply enter a } (Carriage } Return) at the end of each line of text you type (ie. don't let your } system } automatically word wrap) and things should be fine. } } --------------------------------------------------- } ------------------------ } How do I submit/post a message? } ------------------------------- } -------------------------------------------- } } Compose your message offline, then read it a second time! } } Send your message by Email to: "Microscopy-at-MSA.Microscopy.Com" } } As a courtesy to the readers of this list please indicate in the } subject } line of your message a reasonable descriptive title of your } comment/inquiry. } Also preface your description by the conventional abbreviation of the } type } of microscopy you are interested such as LM, IRM, XRM, TEM, SEM, AFM, } STM, } uProbe...... For example, if you are interested in optical microscopy } and } have a question about staining then a Title/Subject line for your } message } might be } } Subject: LM Need help on Staining Cells } } or if you are interested in TEM analysis of dislocations then } } Subject: TEM Dislocation Loop Analysis } } This way individuals can filter out the messages that they may not be } interested in. } } Can I send an Email Attachment? } } Please DO NOT send files as an attachment to the Listserver. } } Many people use email software which displays plain text messages } automatically but requires several manual steps to read and delete or } save } an attachment Others have no capabilities at all for extracting and or } } translating attachments. Please send all messages as plain text. This } will } insure that ALL subscribers to the Listserver can rea ALL messages... } } --------------------------------------------------------------------- } ------ } How do I reply to a message: } ---------------------------- } ----------------------------------------------- } } Compose your message offline, then read it a second time! } } Send a NEW message to "Microscopy-at-MSA.Microscopy.Com" } } DONOT use the REPLY function of your Email Program!! } } If you do this (i.e. use REPLY) then, in most cases, only the person } that } originated the message will see your response. The current } configuration of } the Listserver is such that a message is basically REFLECTED from the } MSA.Microscopy.Com site to all subscribers on this list. the } repackaging of } the message is minimal and not all Email programs will recognize that } the } message reply needs to go back to Microscopy. A good fraction will } reply } back only to the originator. Therefore, if you want everyone to } benefit from } your problem and and/or it's answer you must send it as a message to } "Microscopy-at-MSA.Microscopy.Com" } } ------------------------------- } -------------------------------------------- } What can I post? } ---------------- } ----------------------------------------------------------- } } Basically any question/comment/observation or general } information/announcement which involves microscopy and/or } microanalysis. } This list is not moderated, so it is up to the users to selfpolice } themselves. I generally take an openminded attitude toward questions. } If in } my opinion, it appears that a discussion is straying far off the } limits of } the intent of this forum I will post a note to the group. But I prefer } to } err on the safe side with most messages and allow at least a modicum } of } generally. Clearly, generic questions about Word Processing do not } belong in } this discussion forum, so just use common sense. } } If there is any question in your mind about something you wish to } post, send } it to me first at Zaluzec-at-MSA.Microscopy.Com and I will give you my } opinion/comments. } } ----------------- } ---------------------------------------------------------- } Can I post an Announcement of a Job Opening or a Meeting? } --------------------------------------------------------- } ------------------ } } Yes that falls within the bounds of the subject of this list as long } as it } is related to Microscopy/Microanalysis. } } --------------------------------------- } ------------------------------------ } Can I post my Resume'? } ---------------------- } ----------------------------------------------------- } } No. This forum was not created for that purpose. } } ------------------------------------------------ } --------------------------- } Can I post an Advertisement? } ---------------------------- } ----------------------------------------------- } } No, that does not fit within the bounds of this discussion forum. } } This listserver is not intended to be a Sales mechanism for commerical } } organizations, but rather it is an open discussion area about } microscopy and } microanalysis problems and solutions. If you are an organization and } have } equipment you wish to donate, or sell, for nominal cost (i.e. no } profit) } then this is generally an acceptable posting. If you are not sure then } send } a copy of the announcement in question to Zaluzec-at-MSA.Microscopy.Com } and I } will give you my opinion. An example of this type would be an old } decommissioned instrument which someone is trying to give away for } removal/shipping costs, that would fit within the bounds of the } purposes of } this list. } } If you are a manufacturer, you are always welcome to observe/join in } any } discussion at any times. We do ask that everyone, please refrain from } overt } sales pitches and/or commericalism. If a product which you } produce/sell can } solve a problem or answer a question raised by anyone on this list, } then by } all means feel free to say so. Try to be brief about the product, } state the } simple facts in a few (short) sentences and then offer to continue the } } discussion with any interested parties offline. Alternatively you can } give } sufficient information so that individuals can download/access the } relevant } information.Usually it will be sufficient to just add your phone } number } and/or Email address to the end of your message, and you'll be } contacted by } anyone that is interested. } } Remember, please keep your comments about any product you "sell" to a } minimum. } } It is not out of line to provide your company name, Email address or } WWW } site as part of your signoff/signature line, at the end of ANY message } you } post to this system. } } This Listserver operates on the honor system with respect to to } posting of } advertising, so please respect these simple ground rules. } } If you are interested in using the Internet for Commerical Advertising } of } Microscopy/Microanalysis Related Products/Services, you may wish to } contact } MSA at it's WWW site (http://www.msa.microscopy.com) or the MSA } Business } Office (MSABusinessOffice-at-MSA.Microscopy.Com). These alternative } Internet } services, are provided independently of the Listserver Operation, } which MSA } provides as a FREE service to the WorldWide Microscopy and } Microanalysis } Community. Any funds derived from the above are used to defray the } costs of } running MSA's Internet site. } } ---------------------------- } ----------------------------------------------- } I'm only interested in XYZ Microscopy is there sublist? } ------------------------------------------------------- } -------------------- } } Nope, not here. There are other listservers which deal with specific } topics, } but this one is general. At sometime in the future I may consider } creating } subtopics. But really that is what the SUBJECT line is for. Please use } a } good description for your message! I've learned alot by just } "listening" in } on discussions about topics I know nothing about. You may also find } the same } is true for you. } } ---------------- } ----------------------------------------------------------- } Can I access this list via NewsGroups on the Internet? } ------------------------------------------------------ } --------------------- } } Not any longer. For a time this Listserver automatically forwarded all } } postings to the SciTechnqiue.Microscopy Newsgroup. However, the link } to that } newsgroup has been lost at the receiving end. A link may once again be } } possible in the future, when hardware and software is upgraded. } } --------------------------------------------------------------- } ------------ } I think I need to test the Email, what do I do? } ----------------------------------------------- } ---------------------------- } } First, if you received the instructions, after submitting } instructions, then } your Email program must be working. So no further testing is really } necessary. } } DO NOT send TEST/HELLO messages to Microscopy-at-MSA.Microscopy.Com it } needlessly clutters up the subscribers mailboxes. } } Remember every time you post a message to the Listserver over 2000 } copies } are sent. This not only wastes processing time on my computer, but } gets lots } of people angry at wasted bandwidth. You should also remember some } people } have to pay $$ just to read their Email (think of those subscribers on } } CompuServe, AOL, GENIE, etc....) We have a lot of them, and this } wastes time } &/or money! } } If you really feel you must test your Email Package or think you need } help } then send a message to: } } Zaluzec-at-MSA.Microscopy.Com. } } He's sometimes a patient soul, who at times has been known to read his } Email } and occasionally answers questions. Especially, late at night. } } -------------------------------------------------------------- } ------------- } Is there a Digest Mode? } ----------------------- } ---------------------------------------------------- } } Digest Mode is not available on the present server. It is an option } which } will be added in the future. } } Digest Mode compresses all message from a single day into one long } message } which is then sent to all subscribers. It is a nice option, and I } intend to } get it running eventually. } } -------------------------- } ------------------------------------------------- } When I send a message I get (sometime many) Bad/Bounced Messages back } why? } ---- } ----------------------------------------------------------------------- } } Because this list acts as a reflector, when message delivery fails and } is } returned to the system it goes to the originator of the message. Due } to the } large number of users on this list it is ineveitable that some systems } will } go offline, are down for maintenance, or simply shut down by the user. } The } mailserver tries to deliver messages several times before it gives up. } } ---------------------------------------------------------------------------- } } Can I get a list of the Subscribers: } ------------------------------------ } --------------------------------------- } } No. I do not release that information. There are in excess of 3000 } subscribers from more than 40 countries to this list. } } ----------------------------------------------------- } ---------------------- } Is there an Archive? } -------------------- } ------------------------------------------------------- } } Yes. You can access the monthly archive at the WWW site: } http://www.msa.microscopy.com/MicroscopyListserver/Micro } copyArchives.html } } } --------------------------------------------------------------------------- } } What is the subscription fee? } ----------------------------- } ---------------------------------------------- } } The Listserver is a free service brought to you by MSA there are no } charges } for subscription/use. I'd even be willing to guarentee that you will } get } back at least whatever you put in. If you feel generous, and you } happen to } see your Friendly Neighborhood SysOp at a Microscopy Conference you } can } always buy him a beer or two. He's usually easy to recognize, and his } arm } twists easily. } } -------------- } ------------------------------------------------------------- } Who do I contact with problems? } ------------------------------- } -------------------------------------------- } } God-at-Garden.Eden.Earth.Com } } ------------------------- } -------------------------------------------------- } Who Runs this? } -------------- } ------------------------------------------------------------- } } This listserver is run by Nestor J. Zaluzec } (Zaluzec-at-MSA.Microscopy.Com) It } has been & continues to be run/administered/babied mostly in his spare } time } and usually late in the evening or earlyhours of the morning. } } ------------------------------------------------------------- } -------------- } Special Announcement/News } ------------------------- } -------------------------------------------------- } } At the 1995 Winter Council Meeting, the Microscopy Society of America } (MSA) } approved a proposal to support this server as well as other } telecommunications options as a FREE Service to ALL } persons/organizations } worldwide who are involved or interested in Microscopy and/or } Microanalysis. } } Thanks to this support, the listserver will be upgrading the hardware } software and services for use by Microscopist's/Microanalysts } Worldwide. } } ---------- } ----------------------------------------------------------------- } Is there an FTP and/or WWW site? } -------------------------------- } ------------------------------------------- } } Yes, both an FTP and WWW sites exist here are the addresses: } } WWW http://www.msa.microscopy.com } } FTP ftp.msa.microscopy.com (Anonymous Login Enabled) } } ********************************* } Last Updated April 15, 1997 } Nestor J. Zaluzec } Your Friendly Neighborhood SysOp. } ********************************* } End of File } *********************************
} } The Company: } } Samsung Austin in Austin, Texas wants to offer you more than a job. We want } to offer you the chance to develop a career. Becoming involved in a } fast-paced start-up operation is both exciting and challenging and we look } for employees who are energetic, flexible, and team-oriented. } Are you willing to go the extra mile? Are you comfortable with change? Are } you interested in great benefits? Are you excited about working in a start-up } environment? Are you interested in a career where you will be an integral } part of a new company? Are you comfortable making decisions that will } influence the development of our corporate culture? } If the answer to these questions is "yes," then Samsung Austin is the company } for you. } For more information visit our web site at www.sas.samsung.com } } } The Position } } We are currently looking to hire an Entry Level TEM Technician. The } requirements for this position is simply an Associate Degree in a technical } major. Experience is preferred but not necessary. The position will involve } shift work. } } If someone know of a person who would consider this position or if you know } of a 2 year college that offers courses on TEM Sample Prep please feel free to contact me. We are going to start interviewing next week so please } get your resume to me quickly. } } Resumes can be faxed or emailed to: } David A. Griffiths } Fax (512)491-1165 } Pager (512)209-4132 } e-mail DGriffiths-at-sas.samsung.com }
Thanks to all for your help! We were able to get a pattern after tweaking the specimen quality.
************************************************************************** Lucille A. Giannuzzi, Ph.D. phone: 407 823-5770 University of Central Florida fax: 407 823-0208 Dept. of Mechanical, Materials, and Aerospace Engineering PO Box 162450 4000 Central Florida Blvd. Orlando, FL 32816-2450 email: lag-at-pegasus.cc.ucf.edu **************************************************************************
Peggy Bisher wrote: ================================================= I need to purchase 300 mesh copper and 1000micron-slotted or hole copper grids in the 2.3 mm size. Does any vendor in the US supply these? I know that Agar does in the UK but I thought it might be faster to purchase them on this side of the pond. Can anybody out there help me? ================== ================================ Because of such little demand for the 2.3 mm size, in North America, they not found as readily here as they are in Europe. However, you can find them in a few mesh sizes on our website under "regular" grids, "micron" style. Other mesh sizes and types are available also but are not yet up on the website. I think that some of the other EM suppliers of consumables in the USA offer 2.3 mm grids as well, or at least they did until recently.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I'd like to examine a sample in my SEM. My concern is that if the beam heats the sample, it may volatile and contaminate my column. I don't have a cold finger. Does the beam heat the sample? What is the temperature of the beam? What sort of elevation in temperature does a sample go through during an examination? What about different accelerating voltages, do they produce beams with different temperatures? Thanks Mark E. Darus
Dear colleagues and calibration standard manufacturers.
A committee within the Dutch Microscopy society is working on the general issue of calibration. In June we made a request on the microscopy listserver. The only helpful item that came up was a reference for the NIST calibration standard SRM484 G (which I already own).
On the Internet there is however a large variety in calibration standard available. It's a pity that so little people have responded therefore one can only assume that they are: 1. not using the standards 2. not interested in obtaining accurate results to satisfy the ever demanding customer 3. not on the Internet 4. don't feel the need to talk to us (were told not to by there boss or were shy) 5. under the impression we're not nice persons 8(
Therefore I see no alternative then to ask you again a few questions. The questions are intended for users of a SEM and the manufacturers of calibration standards but anyone who has some interesting things to say is welcome.
Are you all aware of the writing of Herr Clive Walker on "The Report on the 4th Plenary Meeting of ISO/TC202 Londen 6-9 th May 1997" ??. In this writing there are still some issues that must be dealt with. One issue is calibration but unfortunately a consensus was not reached and the issue remains open. So, let's say I want to apply for a STER-lab certificate; what issues are to be dealt with other than calibration (other than the references given in the NIST calibration routine).
Some questions that come to my mind are: * Standard defining terms used in Micro Beam analysis. * General machine settings ? * Info about the mounting size. * Is the sample 1D or 2D (the NIST standard is a 1Dimension standard, IBSEN has a 2D standard) are they the same in the sense that they can both be used for the same calibration routine (for instance the ASTM E 766-93). * Is it important to know about the difference in Z (atom number). The NIST standard has thin gold lines in a nickel base. Others use a silicon grid. Does the difference in Z also mean that there might be a difference in signal to noise ratio when the same machine offsets are used. * Can you give us some photograph's of the calibration sample at let's say 3 different magnifications, 3 different accelerating voltages and corresponding spotsizes. * Can you give us detailed information about the accuracy of the calibration standard and a normal achievable level of accuracy with an average SEM (knowing it's hard to define a normal SEM) including the methods used for statistical proceedings eg. . * Do you have references from laboratory that have a STERlab certificate(not ISO 9000) or equal and I refer to the new ISO/IEC guide 25 using your calibration standards.
I know of the following calibration standard manufactures: Ernest F. Fullam, Inc. SPI. Energy Beam sciences Inc. NIST. MOXTEK, order at Ted Pella, Inc. TCL (I am not sure if they make standards but they do SEM calibration) IBSEN MAG*I*CAL (National Research Council of Canada)
All info is derived from the Internet using common search engines. The list is probably not complete and I would appreciate it if the list can be made up to date with more info.
It is not my intention to give a rating for the calibration standards that are provided with a large variety in spec's and cost. What I (we) want is to give people who are on the road for ISO/IEC certification some info about the calibration standards and the issues that come along in the process. Therefore I give everybody a equal opportunity to give info about his or her calibration sample making it more easy for us people to make a choice in what calibration standard the most suited is for his or her specific use.
As stated above this request goes to all the known manufacturers on the list and this request will also be posted on the Microscopy list server.
Many thanks in advance,
Gert ten Brink Philips Semiconductor BV. Fysisch analyst postbus 10 9500AA Stadskanaal tel 0599632380 fax 0599632505 e-mail brink-at-skn.sc.philips.com privat asslab-at-xs4all.nl
Ps. all cost made and found reasonable (photo material, stamps etc.) will be refunded. On the other hand: if this leads to a large rise in sales in calibration standards can I give you the number of my bankaccount ?
NIST t.a.v. Thomas E. Gills Gaithersburg, Maryland, MD 20899
Allen R. Sampson Advanced Research Systems St. Charles, Illinois ISO/IEC Guide 25-draft four Recommendations for implementation of ISO Quality Standards in an SEM laboratory Both can be found on the Internet
Your profits are about to increase by 50%, practically overnight! If you are selling any type of product or service, on-line or off-line, this software is an absolute MUST for your business. Please read on...
(To contact us, place an order, and/or view this same information online, please visit our web site: http://205.199.2.39/zr1 ) (You can also enter our online sweepstakes to WIN a Pentium laptop PC!) (Note: Our server is in the process of an upgrade, so if you experience any difficulties accessing our web site, please try again later.)
----------------------------------------------------------------------------- "What is this new software?"
- CHECKS XPRESS by ZR ONE Technologies is a powerful new software program that allows you to accept check payments from your customers by telephone, fax, and internet/e-mail.
----------------------------------------------------------------------------- "Why should I accept check payments by telephone, fax, or internet/e-mail?"
- Simple. INCREASED REVENUES - by up to 50% or more! More and more businesses are adopting this new system of accepting payment from their customers. They have all experienced INCREASED PROFITS by providing a new medium for their non-credit card paying c ustomers to purchase their products and services.
----------------------------------------------------------------------------- "How will Checks Xpress increase my revenues by 50%?"
- As you are probably aware, credit cards invoke IMPULSE SHOPPING - you get the customer when they're "HOT". As you are also probably aware, obtaining a credit card merchant account for a small or home-based business can be very difficult and very expensi ve. Therefore, if your customers have the opportunity to "cool down" because they have to manually write you a check and put it in the mail, statistics prove that your chances of losing the sale increase by more than 50%.
- Also, there are over 70 MILLION consumers and businesses that have checking accounts, but DO NOT have credit cards. So even if your business does accept credit cards, you are missing out on a VERY large segment of would-be customers that want to order y our products or services, but do not have credit cards.
- With Checks Xpress, you now have the same impulse-shopping payment power as accepting credit cards! Your customers can order immediately (when they're "HOT"), you GET PAID IMMEDIATELY, and your customers will receive their orders much faster, too. It's a win-win situation for everyone! Plus, we will show you how to verify that your customer's check is "good for the money" within minutes of receiving it, allowing you to fulfill the order only moments after it is placed. This is truly the same power as a ccepting credit cards - and no more waiting (or hoping) for that check to arrive in the mail!
- Eliminate bad debt, collections, and past-due accounts. With your customer's initial one-time authorization, you simply print out a check each month, PAID IN FULL AND ON TIME!
----------------------------------------------------------------------------- "How does this system work?"
- It's easier than you might think. A customer with MONEY TO SPEND places an order through telephone, fax, or e-mail. They provide you with all of the bank and checking account numbers that appear on their check.
- You simply enter their bank information into the Checks Xpress software and click the "Print Check" button. The Checks Xpress interface was designed to look just like a check, making this program super easy to use, and printing the check from start to f inish in less than 2 minutes!
- Then go to your bank and deposit the check(s), just as you would deposit any other checks. That's it - instant money!
* Note to non-U.S. residents: Checks Xpress cannot be used to debit non-U.S. bank accounts; however, if you're doing business internationally, you can still use Checks Xpress to accept payment from all U.S. based consumers and businesses, provided that yo ur financial institution accepts U.S. checks.
----------------------------------------------------------------------------- "Is it legal to print checks this way? What about my bank - will it accept these printed bank drafts?"
- Absolutely! The following federal regulations make this all possible and 100% legal: "Uniform Commercial Code, Title 1, Section1-201(39) and Title 3, Sections 3-104, 3-401, and 3-403; Code of Federal Regulations, Title 12, Chapter II, Part 210 and Reg ulation J, Federal Reserve Bank, Part 2, Sections 4a-201 to 4a-212. Only verbal agreement is required for authorization. See Romani versus Harris 255 Md. 389."
- With the above federal regulations in place, every U.S. banking institution MUST accept your bank drafts, provided that the checks conform to the standards set forth by the American Bankers Association (ABA). Checks Xpress checks comply 100% with the AB A's standards!
- No special bank account or authorization is required - your current bank account is already able to accept bank drafts! In addition, no signature is required to deposit and process the check (refer to the federal regulations listed above). As long as the customer gives you authorization (verbal or otherwise) to debit his/her checking account, a signature is not mandatory. Checks Xpress will automatically print the required authorization information on the signature line of the check.
- Also, no special magnetic ink toner is required for your printer. Since most banks are using the optical (laser beam) check reading system, ink toner is not an issue. For those banks that might still be using the older, magnetic check reading system, yo ur check will still be processed manually. Either way, your Checks Xpress checks are as good as gold!
----------------------------------------------------------------------------- "What are the features of Checks Xpress?"
- Designed exclusively for Windows 95! Because of its 32-bit design, Checks Xpress gives you the performance and user-friendliness you demand.
- Simple, friendly user interface. No special computer skills required. Checks Xpress was created with the most novice of computer users in mind. In fact, Checks Xpress has been rated as the EASIEST to use check printing program, ever. Installation is a b reeze - you'll be printing checks within minutes!
- Store an unlimited number of your customers' checks on your computer, for archival, future retrieval, and monthly account debiting.
- Automatic check number incrementing feature allows you to automatically maintain sequential check numbers for better record keeping and less data entry.
- Custom signature line feature allows you to customize the information that appears on the check's signature line, or use the built-in authorization information along with a date and time stamping feature.
- Custom printer alignment. Easy alignment feature ensures that Checks Xpress will print perfect checks every time with your laser or inkjet printer.
- Free tech support should you ever run into any difficulties with Checks Xpress.
- Checks Xpress is not to be confused with the accounting or financial software programs available in stores (Quicken, Peachtree Accounting, etc.). Unlike those programs, Checks Xpress prints the special bank codes found along the bottom of every check. T he store-bought programs do not print the special bank codes, and they are designed to only print checks from the same checking account.
----------------------------------------------------------------------------- "What do I need to use Checks Xpress to start cashing checks?"
- Any computer system running Windows95 or Windows NT 3.51/4.0 - A laser or inkjet printer - 3.5" floppy disk drive - 5 MB free hard drive space - A standard checking account
----------------------------------------------------------------------------- "What are the costs involved?"
- The Checks Xpress Software: NOW AVAILABLE FOR ONLY $139.00! Our competitors charge $200, $300, even as much as $500 for check printing software programs of lesser quality!
- Blank checks: Can be purchased from a variety of vendors. Our high quality Security Checks feature built-in watermarks and are ideal for use with Checks Xpress. They are formatted one check per 8.5"x11" page. Prices are listed below on the Order Form.
- Bank fees, service fees, monthly fees: NONE! Unlimited check printing, with no other costs other than blank checks.
*** SPECIAL OFFER *** If you order within 10 days of receiving this offer, you will be eligible for a $40 INSTANT ONLINE REBATE! Your total cost to own Checks Xpress WILL BE ONLY $99.00! We GUARANTEE you will not find a better check printing system for le ss! To qualify for your $40 discount, you will need to provide a valid Instant Online Rebate (IOR) number when you place your order. * IMPORTANT * DO NOT LOSE THIS NUMBER - your valid IOR number is: CXP6410
----------------------------------------------------------------------------- "Does ZR ONE Technologies offer a money-back guarantee?"
- Absolutely! You can try Checks Xpress AT NO RISK! After you have tested Checks Xpress in demo mode to your complete satisfaction, simply contact out technical support department to obtain your valid "unlock code", which will permanentely remove all limi tations. At any time during the first 10 days of ownership, or until ZR ONE Technologies issues your Unlock Code, whichever somes first, SHOULD YOU DECIDE THAT YOU ARE NOT COMPLETELY SATISFIED WITH CHECKS XPRESS, YOU MAY RETURN IT FOR A PROMPT AND COURTEO US REFUND, NO QUESTIONS ASKED.
----------------------------------------------------------------------------- - Unleash the POWER of credit cards with the EASE of cashing checks! - Print MONEY directly from your home or office! - Order NOW while the $40 Instant Online Rebate Offer still lasts!
-------------------------------- cut here ----------------------------------- (You can also place an order online via our web site: http://205.199.2.39/zr1 )
YOUR INFORMATION: Business Name (if applicable):________________________________ First Name: __________________ Last Name: ____________________ Address: ________________________________________________ City, State, Zip, Country: ______________________________ E-mail Address: _____________-at-______________________ Phone Number (optional): ___________________________
SOFTWARE ORDER (check one): ___ Please rush me the Checks Xpress software for only $139.00 ___ ** SPECIAL OFFER ** I am ordering within 10 days of receiving this offer, and I would like to take advantage of the $40 Instant Online Rebate Offer as promised above. Please rush me the Checks Xpress software at the special discounted price of only $9 9.00. My valid Instant Online Rebate (IOR) number is: CXP2262
BLANK CHECKS ORDER (check one): ___ 50 blank Security Checks for $19.00 ___ 100 blank Security Checks for $29.00 ___ 250 blank Security Checks for $49.00 ___ I do not wish to order any blank checks at this time.
SHIPPING & HANDLING OPTIONS (check one): All orders within the U.S. and Canada are shipped via UPS, all others are shipped via USPS. Rates shown below remain the same regardless of order size. ___ UPS Normal Delivery = $5.25 (3-6 days) ___ UPS 3 Day Select = $7.75 ___ UPS 2nd Day Air = $10.50 ___ UPS Next Day Air = $21.00 ___ International (outside U.S.) = $13.25 ___ *Send to e-mail account = $0 (within 48 hours) * Note: The e-mail delivery option is available only for software orders NOT consisting of any additional, non-software items.
$_________ TOTAL CHARGES (including S/H)
Select Payment Type: ___ Personal/Business Check ___ Money Order/Certified Check ___ Visa ___ MasterCard ___ American Express Card Number:_________________________ Expiration: ___ / ___ Name on Card:________________________ Signature:___________________________
CHOOSE A WAY TO SEND YOUR ORDER (#1 or #2) : #1. MAIL: Print this order form and mail along with your payment, payable in U.S. funds to:
ZR ONE Technologies Dept. CXP 467 Saratoga Avenue Suite 1425 San Jose, CA 95129 USA
#2. FAX: Print this order form and either (a.) tape your voided personal/business check below this line, or (b.) fill in your credit card information in the spaces above, and fax your order to: 408-260-7811
ZR ONE Technologies will debit your credit card/checking account for the amount stated above in the TOTAL CHARGES line. * ZR ONE TECHNOLOGIES WILL ASSESS A $25.00 FEE FOR ANY RETURNED CHECKS.
For information about our Authorized Reseller (AR) program, visit: http://205.199.2.39/ZR1/reseller.htm
Thank You for your order! We appreciate your business and we look forward to serving you again in the future.
Copyright (c) 1997 ZR ONE Technologies. All rights reserved.
Dear all, I am interested in this question, I sometimes see evidence of local heating of samples during TEM observation but I have never had much idea about quite how much heat is generated by the electron beam. Does anyone know or know how to find out?
Also, does anyone have any idea why oily blobs (obviously from oil in the vacuum system) sometimes appear on the sample in the very place that you are observing (as opposed to any other place on the sample)?
Thanks
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
} I'd like to examine a sample in my SEM. My concern is that if the beam } heats the sample, it may volatile and contaminate my column. I don't have } a cold finger. Does the beam heat the sample? What is the temperature of } the beam? What sort of elevation in temperature does a sample go through } during an examination? What about different accelerating voltages, do they } produce beams with different temperatures? } Thanks } Mark E. Darus
The beam certainly can heat some specimens to a sufficiently high temperature to cause evaporation. The temperature reached is going to depend on all sorts of things - specimen thermal conductivity, how well specimen is connected to mount, how well mount is connected to stage, beam energy and current, scan speed (versus leaving probe stationary), probe size, etc.
Personally, unless you have got something really nasty - elemental arsenic maybe, or mercury - under normal circumstances, I would say that the volume evaporated is going to be so small that there is no need to worry. Although if you are going to be looking at a volatile specimen for 12 hrs a days, 7 days a week you might want to start thinking about a cold trap.
One thing to try to check is vacuum level - it this doesn't change when examining the potentially volatile specimen, I wouldn't get too concerned.
I work in a hospital lab and our pathologist would like some EM pictures of plts. Does anyone have a procedure for isolating plts. from whole blood and embedding them in Spurrs?
I know prices are high. Must have to do with low volume, lots of labor costs, supply and demand, profit??? Anyway... My "primary" standard is the NIST SRM-484 (last time I looked was in the $700-800 range). I also use NIST tracable sphere suspensions from Duke Scientific. Another "standard" I use for very low magnification is a section from an etched steel rule (from Starret, or equal). The rule does not come with a "pedigree", but is NIST tracable through the manufacturer and is an "industry accepted" measuring device.
Woody White Mcdermott Technology, Inc. http://www.mtiresearch.com/ http://www.geocities.com/capecanaveral/3722
Dear Woody, Do you use NIST-certified magnification standards? I found one from Geller Microanalytical which is $1500. Do you know of other suppliers, perhaps cheaper? Thanks, Melanie Behrens (going ISO as soon as I get all this paperwork done....)
The easist way to isolate platlets is to obtain a 7cc tube of blood in an ACD tube (EDTA will also work), centrifuge for 15 minutes at 100 x g. Remove the supernatant (this is the platlet rich plasma), high speed spin this to form a pellet and process as normal.
Best of Luck, Ed Calomeni Dept. Pathology Medical College of Ohio Toledo, OH 43614-2598 emlab-at-opus.mco.edu
The easist way to isolate platlets is to obtain a 7cc tube of blood in an ACD tube (EDTA will also work), centrifuge for 15 minutes at 100 x g. Remove the supernatant (this is the platlet rich plasma), high speed spin this to form a pellet and process as normal.
Best of Luck, Ed Calomeni Dept. Pathology Medical College of Ohio Toledo, OH 43614-2598 emlab-at-opus.mco.edu
Ian MacLaren wrote: } I am interested in this question, I sometimes see evidence of local } heating of samples during TEM observation but I have never had much } idea about quite how much heat is generated by the electron beam. } Does anyone know or know how to find out?
Dear Ian,
L. W. Hobbs has a contribution entitled "Radiation Effects in Analysis of Inorganic Specimens by TEM" in "Introduction to Analytical Electron Microscopy" edited by J. J. Hren, J. I. Goldstein and D. C. Joy, Plenum Press 1979. In this paper there is on page 441 a section on "Electron-Beam Heating" where you will find the relevant equations.
The sample temperature will depend on beam current, thermal conductivity and specimen geometry. Hobbs mentions that under the worst circumstances it is possible to melt refractory ceramics.
Best wishes, Joergen.
J. B. Bilde-Soerensen Senior Research Scientist, ph. d. Materials Research Department Risoe National Laboratory DK-4000 Roskilde Denmark
Dear all, A few months ago I asked my Professor about heating of samples during TEM observation and he suggested me to read the monograph of L.Reimer: Transmission Electron Microscopy (Springer-Verlag, Berlin, 1984). I have read and I can recommend you as the good source regarding "Beam Temperature".
In response to Ian MacLaren's request, hydrocarbon contamination is resident on EM specimens as a result of preparation and handling techniques, , ambient conditions, and microscope vacuum contamination, although in most cases it has been found that the microscope vacuum is actually quite clean.
Under vacuum conditions, the hydrocarbons are mobile on the specimen surface. As they pass the impingiment point of the electron beam on the specimen, they are essentially polymerized. With the beam focussed at one area on the specimen, as is the case when conducting fine probe microanalysis in a TEM, a carbon cone is generated. On a TEM specimen, the carbon formation is created simultaneously from both specimen surfaces. These carbon formations or "oily blobs" as Ian MacLaren called them preclude both imaging and the acquistion of analytical data.
It has been found that low-energy plasma cleaning of the specimen prior to EM observation, utilizing an oxygen-based process gas, chemically converts the hydrocarbon contamination to CO, CO2 and H2O. This process virtually eliminates the contamination issue without altering the specimen's properties. The resultant is enhanced imaging and analytical data.
Best regards,
Paul E. Fischione E.A. Fischione Instruments, Inc. 9003 Corporate Circle Export, PA 15632 USA Phone 412-325-5444 FAX 412-325-5443 Web site: www.fischione.com
Hi everybody According to Castaing in thesis the temperature rise at the point of irradiation theta in Celsius is=20 approximately as follow:
theta =3D 1.14 IaV/Cd
Where Ia specimen absorbed current in microamp V accelerating voltage in kV C Thermal conductivity (cal/cm sec deg) d electron probe diameter in micronmeter
for example with glass at 10kv and d=3D0.1=B5 and Ia=3D0.1nA rise of temp is about 7 degrees celsius
If somebody interresting I also have a diagram showing temperature rise vs thermal conductivity (about 50k in TIF) Hope that helps.
Eugene, I use Spurr's exclusively for E.M. The stain I use on 0.5 micron sections is Paragon. It is a combination of Toluidine Blue and Basic Fuchsin and is a really beautiful stain. The reference for this is Dr. Frieda Carson's book "Histologic Techniques In Electron Microscopy", published by the American Society of Medical Technology, 1979. Hope this helps. Sharron G. Chism HT (ASCP) Electron Microscopy Lab Harris Methodist Hospital Fort Worth, Texas
Please help me with some recommendations; We are a multi-user, University, primarily biological, microscopy facility used extensively by staff and postgraduates from the Faculties of Science and Agriculture.
For the past ten years we have made extensive use of a steam(286)-driven Kontron Vidas full colour Image Analysis system which, though competent and versatile, was soon dubbed 'user-hostile' by our students.
We now have a very limited budget with which to replace this apparatus. Any new system would need to be Windows-based (to satisfy those students !) and have a high specification of PC hardware which is easily obtained from local suppliers.
I am looking for proven recommendations for software used in similar applications to our own. Further detail can be supplied on request.
I would particularly appreciate recommendations on reasonably priced products via fellow microscopists. Our applications range over particle counting through a great variety of macro and micro area measurement to measuring the black vs white area on dairy cows !!
Commercial responses should be mailed directly to me in the spirit of correct net protocol.
Tony Bruton Head, Centre for Electron Microscopy University of Natal, Pietermaritzburg. KwaZulu-Natal, South Africa Tel: +27 331 2605155 Fax: +27 331 2605776 E-mail: bruton-at-emu.unp.ac.za
FALL 1997 COURSE ANNOUNCEMENT - Transmission Electron Microscopy (BIO. 221-V2)
NASSAU COMMUNITY COLLEGE
A fifteen week, fall 1997 semester, course in Biological Transmission Electron Microscopy is being offered by the Biology Department of Nassau Community College. This is a 4 credit course offered ONE EVENING PER WEEK, Thursdays, starting at 5:30pm. Classes will begin on Sept. 4 and end on Dec. 18, 1997.
This is a "hands-on" course emphasizing biological specimen preparation, ultra-thin sectioning involving block trimming, glass knifemaking and operation of the ultramicrotomes (Sorvall MT-2B and LKB Ultrotome III), thick and ultra-thin section mounting and contrast staining (UA and Pb citrate), grid support films (formvar, carbon), student operation of the TEM (Hitachi HS-8, Philips EM 300) and production of electron micrographs through the process of black & white photography, and electron micrograph analysis. Students will work on a chosen sample(s) with the goal of producing a portfolio of high quality TEM photomicrographs of that sample(s).
The course is widely transferrable and the cost per credit is reasonable at $84 per credit.
More information about the Bio-Imaging Center at NCC, course descriptions and syllabi, and the beginnings of a student gallery of EM photomicrographs is available at our web site. The URL is {http://www.sunynassau.edu/webpages/biology/becks.htm} .
For those without www access, the catalog description is specified below. If you have further questions, you should e-mail me directly at the address below.
Interested individuals should register early (prior to Aug. 15) since the course is limited to a total enrollment of ten (10) students.
Questions regarding the actual registration process can be directed to our registrar at (516) 572-7355. ________________________________________________________________________________
CATALOG DESCRIPTION BIO 221: Transmission Electron Microscopy -- 4 credits Prerequisites: BIO 109-110 or equivalent, CHE 151-152 or equivalent. An introduction to the basic principles of transmission electron microscopy including tissue preparation, microscope (TEM) operation, black & white photography, and micrograph interpretation. The entire laboratory is devoted to the development of skills and preparative techniques involved with the operation of an actual transmission electron microscope. (3 lecture, 3 laboratory hours). Laboratory fee applies. ________________________________________________________________________________
Stephen J. Beck Associate Professor Bio-Imaging Center/Electron Microscopy Department of Biology Nassau Community College Garden City, NY 11530 Voice Mail: (516) 572-7829 Email: {becks-at-sunynassau.edu} URL: {http://www.sunynassau.edu/webpages/biology/becks.htm}
Dear All, A student, as part of an electron microscopy project, has ferrosilicon samples from powder up to 2mm diameter. After heat treating, he wants to chemically polish the surface to remove contamination. Does anyone know of a chemical polish that would do this? Preferably it should polish not etch the surface. Thanks for any suggestions. Mike
Michael J Witcomb PhD Electron Microscope Unit University of the Witwatersrand Private Bag 3 WITS 2050 South Africa
I am doing in situ hybridization on parafin embedded and sectined plant material using a DIG labelled probe. However, I find that the binds non-specifically to walls and cytoplasm. I have tested the anti-dig AB and the do not show any non-specific binding.
Does someone have an idear on how to block non-specific binding of the probe prior to hybridization without damaging the DNA (or RNA) in the sections?
Peggy Bisher wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Dear Newsgroup: } } I need to purchase 300 mesh copper and 1000micron-slotted or hole copper } grids in the 2.3 mm size. } Does any vendor in the US supply these? I know that Agar does in the UK but } I thought it might be faster to purchase them on this side of the pond. Can } anybody out there help me? Thanks so much. } } Cheers, Peggy Bisher. } } NEC Research Institute } 4 Independence Way } Princeton, NJ 08540. } Tel.: (609) 951-2629 } Fax: (609) 951-2496 } e-mail: peggy-at-research.nj.nec.com
Peggy, We have the following 2.3 mm copper grids in stock. If any are of interest, please e-mail me and I will get you pricing:
Dear Mark, Others have answered the other questions you asked, so I'll just tackle these two.
} What is the temperature of } the beam?
The beam is far from equilibrium, so such concepts as temperature don't really apply, but for a monatomic gas, E = 3/2kT, so for a "gas" of electrons, you can use the same equation. If you had a gas of hot electrons in a chamber with a small hole in one wall, there would be a stream of electrons emerging from the hole whose average energy would be 3/2kT. This is not exactly a mono-energetic electron electron beam, but the same concept can be applied. 1 eV is about 10^4 K, so a 10 keV beam has a "temperature" of about 10^8 K.
} What about different accelerating voltages, do they } produce beams with different temperatures?
From the considerations above, yes.
The connection between the very high temperature of the beam (about that of a stellar interior) and the heating of the specimen is through the energy deposited in the specimen as the electrons are slowed to a stop. In the SEM each electron is stopped in the specimen, so all the energy is converted to heat (except that used in the produc- tion of secondary electrons, taken away by backscattered electrons, etc.). A 10 keV electron has a range of 0.28 mg/cm^2, or--since carbon has a density of ~2 g/cm^3--~1.4 micro meter. Thus, all the energy is deposi- ted in a thin layer near the surface. As others have said, the final specimen temperature depends on how this heat is dissipated. Yours, Bill Tivol
Yes, I have a procedure for what you want. We are experimenting with the following process ...so far so good! (About 5 - 7 mls of blood is collected in an EDTA tube.) 1. Centrifuge blood sample at 1,300 rpm for 10 minutes. 2. Draw off half of the plasma and discard. 3. Centrifuge again at 1,300 rpm for 10 minutes 4. Draw off plasma leaving a 2mm layer over the cells. Be VERY careful not to disturb the cells. 5. Replace the plasma with an equal amount of 2.5% buffered glutaraldehyde. 6. Place this in the 'fridge for at least 4 hours ... overnight is ok. 7. Carefully draw off the fixative and discard. 8. Remove the button of cells with a sharp applicator stick, and place in a dissecting dish. (Try not to break the button too much ... you can probably get it out in two pieces.) 9. Rinse the cells with fixative and remove as much of the red cells as possible with a razor blade. 10. Dice the button into 1mm squares and process as you would tissue. We go through Osmium, graduated alcohols and eventually embedding in Spurr's.
The only difference in this process and our routine buffy coat is the first three steps. We usually centrifuge at 3,000 rpm for 10 minutes and skip steps #2 and #3 for leukocyte study. We have found that the faster rpm can damage platelets so we've come up with this ... see what you think.
Sharron G. Chism HT (ASCP) Electron Microscopy Lab Harris Methodist Hospital Fort Worth, Texas
I work in a hospital lab and our pathologist would like some EM pictures of plts. Does anyone have a procedure for isolating plts. from whole blood and embedding them in Spurrs?
Just a follow up to Paul F.s comment. I would disagree with Paul's statement saying..
"although in most cases it has been found that the microscope vacuum is actually quite clean".
I've done some pretty extensive work on this topic for more years than I'd like to admit to and can show that contamination is also derived from the microscope "vacuum". I've worked with microscopes operating from 10**-5 to 10**-10 torr. Cleaning a specimen with reactive gas plasma minimizes the initial specimen borne components. But if a specimen is left in even a relative modern microscope over night (~ 10**-7 to 10**-8) the contamination effects can return albeit at a reduced level. Subsequent retreatment of the specimen with a plasma will remove this but if you leave it sitting in the microscope it will eventually return.
Stop by the poster session at the Microscopy & Microanalysis 97 meeting in Cleveland and I'll be glad to fill you in.
I recently went through an extensive evaluation of several systems to replace our Kontron system. We examined systems for Mac, Unix and PC systems and determined that a software package called Optimas is the best value for the money. It is a PC based system that works well with Win95 or Win NT (or Win 3.11 for that matter!) The software has many pre defined macros which may help in your application, but also has a very powerful, vector based, C-like language which is fairly easy to use. The support is top notch with and excellent web page: http://www.optimas.com which gives really good support and their phone support is also very good. The company is located in Washington state and our local vendor sold a single license of the software for $3995. We bought our own frame grabber and computer. If you would like to contact me off line to ask further questions, my email is David_Bell-at-millipore.com and my number is (617) 533-2108.
I am in no way connected to Optimas or any Optimas reseller, I am just a very satisfied user.
We are preparing to examine dairy products for protein, fat and starch. If anyone has had experience with this application using a Biorad krypton-argon laser and could give any suggestion on technique and suggest the appropriate flourochromes it would be greatly appreticated.
William R. McManus Electron Microscopy Facility Department of Biology Utah State University Logan UT 84322-5305 1-801-797-1920
Some principles I think are important. Stay away from systems that require proprietary hardware boards for the software to run. These systems tend to become obsolete quickly or are expensive to upgrade. The less expensive image analysis software programs tend to be easy to use but lack the flexibility and power when confronted with a difficult problem. For overall cost and performance, I think PC based systems are the best. My lab has chosen Optimas software (runs under Win95 or NT) as the main image analysis platform, and sofar it has been able to do everthing we require.
Regards,
John J. Turek, Ph.D. Associate Professor Director, Electron Microscopy Laboratory and Core Laboratory for Image Analysis and Multidimensional Applications (CRISTAL) Department of Basic Medical Sciences 1246 Lynn Hall, G193C Purdue University W. Lafayette, IN 47907-1246 Phone: 765-494-5854 Fax: 765-494-0781 Email: jjt-at-vet.purdue.edu
On Thu, 31 Jul 1997 David_Bell-at-Millipore.com wrote:
} I recently went through an extensive evaluation of several systems to } replace our Kontron system. } We examined systems for Mac, Unix and PC systems and determined that a } software package called Optimas is the best value for the money. It is a } PC based system that works well with Win95 or Win NT (or Win 3.11 for that } matter!) The software has many pre defined macros which may help in your } application, but also has a very powerful, vector based, C-like language } which is fairly easy to use.
We did the same and decided on Visilog in part because it uses real C-code and includes a C-interpreter which aids programming. All the other stuff too but the price is a bit more.
Talking about long term storage of tissue reminds of one of my favorite sayings - I always reserve the right to be wrong! - (But I will do my best).
Unfortunately I have not real hard data on tissue storage. This would take an immense amount of time to accumulate. But over many, many years, the following have proven to be good. After glutaraldehyde the tissue remanins quite permeable. Fluids and material may flow in both directions*. Some lipid has been lost*. This is evident when mycobacterium a prefixed with glut and malachite green, and lipid containing filaments which perhaps carry antigen, remain visible. The malachite prevents lipid loss at the outset*. Immediate, superfast, fixation and processing of tissue results in the most "brilliant" of sections and intact cytoplasm. This comes into evidence when one does pathological tissue TEM investigation - wet tissue to paper micrographs in 5.5 hours. I have done this, and the results are astonishing. But we cannot indulge in this. Frequently we must store for weeks, months. This was the case when I worked at a research institution which had to store lung tissue. We stored it in 0.1M buffer after fixation for 4 hours in glut. And we stored in in 0.2M buffer on the assumption (Note: assumption is the mother of all screw-ups) that the increased hypertonicity of the buffer would prevent exodus through membranes. We felt that this improved storage conditions, but we did not do a systematic study, or spend much time at very high TEM magnifications. But we did end up adopting that strategy. Osmium fixes lipids and some proteins*. At a later date I started walking tissue through the osmium step and then storing for long term in buffer. I felt that this was quite an improvement. I have no comparative data. If asked how I would store tissue today, my answer would be to carefully fix it with phosphate buffer and fresh glut, quick rinse it, and refix it in osmium, constantly keeping the tissue in motion, and store it in phosphate buffer in the refrigerator, tightly capped. Storage in alcohol is not advisable, as the alcohol can dislodge the osmium. We see this when the alcohol turns brownish. Long term storage in glut brings up the question of the polymerization and degredation of glut over time*. What effect would this have on the tissue? To really aquire an answer one would have to do a "blind study". That is, the microscopist would have to be required to take the micrographs and then sort them according to their storage conditions without the benefit of any ID data sheet. *Denotes references available for these statements. Bye, Hope to see you all in Cleveland, Hildy
If you worry about cooking a specimen the following guide lines may be of=
assistance.
1. Use a low kV, the lowest you can use with comfort 2 to 5 is possible with most conventional instruments although I have looked at uncoated photoresist at 200v with Lab6. FEG makes the job too easy. 2. Use a low emission current about 20- 50uA with W 3. Use a small spot size, not for resolution but as a safety device.=
4. Set the stage so that when you switch on the beam you are NOT on the specimen 5. Set up off the specimen material and when on the material make yo= ur adjustments about a screen width away from the area required. 6. Only move the beam at the last moment, do not move the stage as this may change the Z .You need a fairly flat specimen or a good depth of=
field to be successful here. 7. Practice focussing and stigmating then press photo and then move the specimen the known amount with a X or Y beam shift. Most instruments=
pause between the photo button press and actually starting the scan. 8. In my experience some specimens will be damaged or contaminated s= o they look different after even one additional scan. This one scan photo method is about a pure an image as you can get.
1 to 3 above are the safety features, cut down the kV and cut down the be= am current and you save your specimen. Keep the kV off and they last even longer :-)
I visit on average one SEM laboratory per week throughout the year and it=
was initially quite a shock to see that very very few really know how good/bad their microscope calibration is!
Coming from the direction where as a TEM engineer I calibrated all of the=
microscopes that I attended once each year, in my teaching I carry this practise over to the SEM. I routinely carry out SEM resolution, magnification calibration and contamination rate tests on the instruments=
that I use. At first I tried drift rate tests too but the results came a= s a shock!
Resolution - most instruments are set up incorrectly. The electron gun i= s always in economy mode i.e. the filament is too far from the cathode to enable spec resolution to be attained. Correct this problem or tune the gun further and it is good to see how many old instruments are capable of=
beating their spec resolution. I use my well know sputtered gold on late= x spheres for this test.
Magnification Calibration - Most instruments are within the standard I fe= el is respectable which is plus or minus 10% of the readout with no more tha= n a 5% error between X and Y directions. What people fail to recognise is that different spot sizes on the same area at the same magnification provide different calibration values. Typical is a ten turn potentiomete= r on old Hitachi instruments 2 turns give a 5% change in calibration. Peopl= e do not seem to recognise that if you change the focus after some other adjustment you have just changed the effective working distance and therefore the magnification. I use an Agar TEM carbon grating replica a cross grating of 2160 line per millimetre. I prefer this specimen as it also makes a very good demonstration specimen on the effect of kV on imag= e form. See "Working With A SEM" S.K. Chapman ISBN 0 850770 93 9. It is m= y experience that on some SEM the magnification calibration is very good at=
certain kV, but bad on others. Machines also seem good at certain WD but=
not at others. In courses each student measures each picture and we have= a spread of 4 to 7% amongst them! It is not that easy to calibrate a SEM!
Contamination Rate - all this comment about oil and specimen damage, is n= ot contamination a cracking of vapours within the vacuum by the heat of the beam on a surface. Is it not hydrocarbons and silicons being deposited hence the low signal level dark lines or rectangles? SEM contamination rate is very much specimen dependant but by taking a constant approach th= is may be a useful test. I use sputter coated latex spheres the specimen being in the microscope one hour prior to the test. A typical rough toug= h microscope used without any care gives 10nm/min over my 20 minute test period. Under similar (emphasise similar) conditions a well kept air locked instrument will come down to 2.5nm/min. Add a cold finger around the final lens similar to that used in a cryo system and you are down to {1.5nm/min.
Drift Rate - I thought SEM stages were lousy however testing a good numbe= r of instruments over a wide price range I found that over a twenty minute period the amount of drift was less than the instruments resolution, in other words the sample did not move. If it did I always found an earth problem not a stage drift problem. I no longer bother with this test unless I have a worry about a particular stage stability. =
Most of my work has been on run of the mill instruments with the best results from the modern twin detector FEG systems. In these instruments = a good cold finger sitting around the final lens is the difference between good and amazing results - contamination IS the killer of high resolution=
If you worry about cooking a specimen the following guide lines may be of=
assistance.
1. Use a low kV, the lowest you can use with comfort 2 to 5 is possible with most conventional instruments although I have looked at uncoated photoresist at 200v with Lab6. FEG makes the job too easy. 2. Use a low emission current about 20- 50uA with W 3. Use a small spot size, not for resolution but as a safety device.=
4. Set the stage so that when you switch on the beam you are NOT on the specimen 5. Set up off the specimen material and when on the material make yo= ur adjustments about a screen width away from the area required. 6. Only move the beam at the last moment, do not move the stage as this may change the Z .You need a fairly flat specimen or a good depth of=
field to be successful here. 7. Practice focussing and stigmating then press photo and then move the specimen the known amount with a X or Y beam shift. Most instruments=
pause between the photo button press and actually starting the scan. 8. In my experience some specimens will be damaged or contaminated s= o they look different after even one additional scan. This one scan photo method is about a pure an image as you can get.
1 to 3 above are the safety features, cut down the kV and cut down the be= am current and you save your specimen. Keep the kV off and they last even longer :-)
I visit on average one SEM laboratory per week throughout the year and it=
was initially quite a shock to see that very very few really know how good/bad their microscope calibration is!
Coming from the direction where as a TEM engineer I calibrated all of the=
microscopes that I attended once each year, in my teaching I carry this practise over to the SEM. I routinely carry out SEM resolution, magnification calibration and contamination rate tests on the instruments=
that I use. At first I tried drift rate tests too but the results came a= s a shock!
Resolution - most instruments are set up incorrectly. The electron gun i= s always in economy mode i.e. the filament is too far from the cathode to enable spec resolution to be attained. Correct this problem or tune the gun further and it is good to see how many old instruments are capable of=
beating their spec resolution. I use my well know sputtered gold on late= x spheres for this test.
Magnification Calibration - Most instruments are within the standard I fe= el is respectable which is plus or minus 10% of the readout with no more tha= n a 5% error between X and Y directions. What people fail to recognise is that different spot sizes on the same area at the same magnification provide different calibration values. Typical is a ten turn potentiomete= r on old Hitachi instruments 2 turns give a 5% change in calibration. Peopl= e do not seem to recognise that if you change the focus after some other adjustment you have just changed the effective working distance and therefore the magnification. I use an Agar TEM carbon grating replica a cross grating of 2160 line per millimetre. I prefer this specimen as it also makes a very good demonstration specimen on the effect of kV on imag= e form. See "Working With A SEM" S.K. Chapman ISBN 0 850770 93 9. It is m= y experience that on some SEM the magnification calibration is very good at=
certain kV, but bad on others. Machines also seem good at certain WD but=
not at others. In courses each student measures each picture and we have= a spread of 4 to 7% amongst them! It is not that easy to calibrate a SEM!
Contamination Rate - all this comment about oil and specimen damage, is n= ot contamination a cracking of vapours within the vacuum by the heat of the beam on a surface. Is it not hydrocarbons and silicons being deposited hence the low signal level dark lines or rectangles? SEM contamination rate is very much specimen dependant but by taking a constant approach th= is may be a useful test. I use sputter coated latex spheres the specimen being in the microscope one hour prior to the test. A typical rough toug= h microscope used without any care gives 10nm/min over my 20 minute test period. Under similar (emphasise similar) conditions a well kept air locked instrument will come down to 2.5nm/min. Add a cold finger around the final lens similar to that used in a cryo system and you are down to {1.5nm/min.
Drift Rate - I thought SEM stages were lousy however testing a good numbe= r of instruments over a wide price range I found that over a twenty minute period the amount of drift was less than the instruments resolution, in other words the sample did not move. If it did I always found an earth problem not a stage drift problem. I no longer bother with this test unless I have a worry about a particular stage stability. =
Most of my work has been on run of the mill instruments with the best results from the modern twin detector FEG systems. In these instruments = a good cold finger sitting around the final lens is the difference between good and amazing results - contamination IS the killer of high resolution=
Greetings, I inherited a Lietz Orthomat Camera system. It has the controler and the camera part. Alas, it says on the box that it is broken and the estimate for repair made in 1986 was $1000 US dollars. This item is not quite old enough to be a "collectors item" but when these function, they are very very good. I would hate to throw this away. I was just wondering if there were perhaps someone who could use the parts? Or who might be able to fix the camera and use it? Thanks, Tobias
We saw the same thing in our XPS system operating in the low 10E-9 to high 10E-10 Torr range when we did our Contamination study that we presented at the Spring MRS97 meeting. We wanted to measure very small quantities of HC's on the surface and wanted to know if the vacuum was contributing. (We were trying to find our minimum detectability limit.) We left the sample overnight after sputter cleaning to a fresh, i.e. no C peaks, and ran the XPS first thing in the morning. A significant surface C peak was present. Of course, any good RGA will tell you how much HC's you have in a vacuum system. -Scott Walck
} } I've done some pretty extensive work on this topic for more years } than I'd like to admit to and can show that } contamination is also derived from the microscope "vacuum". I've worked } with microscopes operating from 10**-5 to 10**-10 torr. Cleaning a } specimen with reactive gas plasma minimizes the initial specimen borne } components. But } if a specimen is left in even a relative modern microscope } over night (~ 10**-7 to 10**-8) the contamination effects can return albeit at } a reduced level. Subsequent retreatment of the specimen with } a plasma will remove this but if you leave it sitting in } the microscope it will eventually return. } } } Stop by the poster session at the Microscopy & Microanalysis 97 } meeting in Cleveland and I'll be glad to fill you in. } } } Nestor Zaluzec } } Your Friendly Neighborhood SysOp. } }
The question of thermal effect caused by electron beam in SEM and X-Ray microanalysis was a subject of studies by Dr. M.N. Filippov in his Doctor of Science Dissertation devoted to the microprobe analysis of unstable samples.
It was found that the theoretical estimation of the overheating of the sample may be expressed as DT = 7.8* Io*Eo*ro/lambda/(ro*do+0.13*Eo^1.7), where Io is the probe current (MickroAm), Eo - electron energy (keV), ro - sample density (g/.sm^3), lambda is the heat conductivity of the sample(Wt/cm/K). DT -s the sample overheating (in K). In his works Dr. filippov also derived the equation to estimate the time, required for the achieving the overheating temperature
t = 2.5E-7*(c*ro/lambda)*(0.5d*do+6.4E-2*(Eo^1.7)/ro)^2. Here t is time in sec, c is the specific heat capasity (J/g/K).
As a consequense of this equations, it was found (and cofirmed experimentally) that the for a lot of samples which suffer from the electron beam induced overheating, the maximum overheating is at about 25-30 kV. If you increase the beam energy, despite the fact that you are starting to pump more energy to the sample, you are also increasing the dissipation surface, and thus, the overheating of the sample often at 50 kV is smaller than the one at 30 kV.
More detailed information might be obtained from Dr. Filippov. (As far as I know his E-Mail is fil-at-pel157a.phys.msu.su Hopefully, this may be usefull.
E-Mail : _ . Nick Kinaev ,~' (_|\Centre for Electron Microscopy(CMM) nick-at-mama.minmet.uq.oz.au ,-' \ The University of Queensland Ph. home : +61 7 3279 4771 ( * {----Brisbane Ph. Dept:+61 7 3365 3743 \ __ / Qld 4072 Fax: +61 7 365 3888 \,~' "\__/ Australia
Bo Johansen laments: } I am doing in situ hybridization on parafin embedded and sectined plant } material using a DIG labelled probe. However, I find that the binds } non-specifically to walls and cytoplasm. I have tested the anti-dig AB } and the do not show any non-specific binding.
Bo binding to cell walls is just one of the joys of working with plant material :-). If you are working with RNA probes try adding tRNA to your hyb buffer. Might also be worth trying things like a "blotto" pre-hyb step similar to membrane hybs.
contact me if you want to talk about this some more.
--
Daryl Webb (dwebb-at-waite.adelaide.edu.au) Dept. of Plant Science, Waite Institute University of Adelaide, Glen Osmond S.A. 5064 Australia. Voice:61_8 8303 7426 Fax:61_8 8303 7102
James Martin wrote: ==================================================== I am looking for information about the ISO 9001 standard with regard to the operation of FT-IR microscopy and SEM systems for materials analysis, and would very much appreciate communicating with list members who have practical experience in this area. ===================================================== Our analytical services laboratory is accredited by the American Association for Laboratory Accreditation (A2LA) to the standard of ISO Guide 25. While on the one hand, it seems like there is a paperwork requirement the describes virtually everything, and while that is at times frustrating, I am quite confident that we have a laboratory running on a far higher level since everyone is much more accountable. While in a sense it does add to our costs, the "cost of rework", that is, the cost of doing samples over again because they were not done right the first time, has gone down more than enough to compensate.
You can contact A2LA directly at the following:
American Association for Laboratory Accreditation 656 Quince Orchard Rd. #620 Gaithersburg, MD 20878-1409 301-670-1377, FAX 301-869-1495 http://www.a2la.org/
A2LA has been accrediting EM and LM laboratories under the discipline "Chemical Analysis" and subgroup "Microscopy". I would imagine that the extension from EM/LM labs to FT/IR microscopy would not be a very great leap.
We have no connection to A2LA except as being one of their accredited laboratories. A satisfied customer, in other words.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
James Martin wrote: ==================================================== I am looking for information about the ISO 9001 standard with regard to the operation of FT-IR microscopy and SEM systems for materials analysis, and would very much appreciate communicating with list members who have practical experience in this area. ===================================================== Our analytical services laboratory is accredited by the American Association for Laboratory Accreditation (A2LA) to the standard of ISO Guide 25. While on the one hand, it seems like there is a paperwork requirement the describes virtually everything, and while that is at times frustrating, I am quite confident that we have a laboratory running on a far higher level since everyone is much more accountable. While in a sense it does add to our costs, the "cost of rework", that is, the cost of doing samples over again because they were not done right the first time, has gone down more than enough to compensate.
You can contact A2LA directly at the following:
American Association for Laboratory Accreditation 656 Quince Orchard Rd. #620 Gaithersburg, MD 20878-1409 301-670-1377, FAX 301-869-1495 http://www.a2la.org/
A2LA has been accrediting EM and LM laboratories under the discipline "Chemical Analysis" and subgroup "Microscopy". I would imagine that the extension from EM/LM labs to FT/IR microscopy would not be a very great leap.
We have no connection to A2LA except as being one of their accredited laboratories. A satisfied customer, in other words.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I would appreciate any comments regarding charge back fees for SEM and TEM services. Specifically, I would like to know how the basis for the charges are derived. The rates listed in the Tech Forum varied so much I was wondering if anyone has worked out a formula on whether to charge per sample or per hour. We are trying to compare our rates with those from other facilities and would really appreciate comments, suggestions etc.
I don't ever remember having spoken to you regarding Visilog. By the way, Optimas does not generate C code in their recorder that's why they use the expression C like, actually it is a language called ALI which is not compatible to Visual C++ in any way.
You say you don't have any interest in promoting their product but you sure sound like it.
For your information, if you had looked at Visilog you would have noticed the following features:
- much wider selection of imaging algorithms than other Windows based sofware.
- wide selection of frame grabbers, Matrox Meteor, Pulsar and now including support for the new Matrox Genesis board using the C80. ALso COreco's TCI, Integral Flashpoint and ITI IC PCI plus new drivers coming for Data Translation and EDT.
- support for NT, Win95 and all Unix workstations.
- Real time morphology using the New Matrox Genesis C80. Pentium Pro's using WindowsNT do not come close to the power of this processor. No other off the shelf software supports dsp based processors like Visilog does.
- The ability to process floating point images for images which are greater than 8 bits per pixel.
- a powerfull c interpreter which generates real C code compatible with Visual Basic and VIsual C++.
- C interpreter for generating low level code and creating stand alone applications.
- Outstanding support and customer service.
- Run time version for as low as 1000.00
- 3 versions of the software starting at 1,000.00$
- On site training and consulting.
- A big and loyal installation base in the US, Europe and Japan,
- new easy to use user interface.
By the way, we will be at the Microscopy show in Cleveland, stop by and I'll give you a good demo. You can also see our new 3d Reconstruction package running on Windows NT.
Regards,
At 02:32 PM 7/31/97 -0400, David_Bell-at-Millipore.com wrote:
} I was wondering if anyone has worked out a formula on whether to charge per } sample or per hour.
It depends on what the lab does.
From the point of view of your "customers" the per sample rates are preferable because then their costs are predictable. It's also easier for you to do the bookkeeping, since all you have to do is count the samples.
On the other hand, if you do non-routine work, there is no way you can do a per sample pricing and come out fair.
As we do a mixture of routine and non-routine stuff, we have a mixed price system. For the routine analyses, we have a per sample rate based on the average time it takes us to handle the sample. In our catalog, we have specified EXACTLY what is included in these routines. Anything that differs from these standard procedures at all, goes on the hourly rate. When we apply the hourly rate, our customers (all internal) have the opportunity of specifying an upper threshold value. When we see that the analysis is going to be more expensive than this value, we call them up and ask them if we should continue. If they say no, then we charge them for the time we spent and give them the results obtained so far (if any).
Hello All, Has any-one any experience with Boehringer Mannheim's antibody to GABAA Receptor alpha chain, specifically whether it cross reacts with rat alpha chain?
Does anyone know of any other available antibodies to GABAA Receptor alpha subunits? I've checked all the usual commercial sources I could think of, with no luck, and would appreciate any suggestions.
Thanks Sharon
Dr. Sharon Miksys Department of Pharmacology University of Toronto 1 King's College Circle Toronto, Ontario Canada, M5S 1A8 Tel: (416) 978-4082 Fax: (416) 978-6395 Email: s.miksys-at-utoronto.ca
For what its worth, I share many of the same concerns with respect to systems that require proprietary hardware. There are many IA systems that will perform a variety of tasks. There are also a wide range of levels of sophistication in many of the IA systems offered to date. I suppose the best choice is determined by the type of work you will be doing, i.e. if you will routinely perform a similar task day in and day out, then a turn key system is probably the best bet, otherwise, you need a system with versatility, and ease of use, key word being EASE OF USE.
In general most IA systems all offer a platform of similar operations, upon these, are added some features that make their software "different" from the pack. Many IA systems have a sleek, and hi tech look, but behind the scenes they are doing many of the same operations as the next IA system. To me the key point (assuming the system is full featured of course) of any IA system is ease of use, ease of programming, ease of program modification
If you are going to be faced with a wide variety of applications, select a system that will be able to handle sample variation during analysis, and that you will intuitively be able to run without spending months trying to learn a programming language. Most of us are not programmers, therefore, ease of programming is paramount. Most systems I have seen allow you to write a sophisticated macros, but they often do not easily allow you to fine tune, or tweak your program without having a good working knowledge of their programming language. In this light, I feel a good deal of attention should be placed on how easy it is to write and modify macros.
Two systems to date that I believe are the easiest to program and modify: On the high end, Kontron KS400, and on the low end Mediacybernetics Image Pro Plus. These two IA programs offer a broad spectrum of applications, and have a good support team.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Tony:
Some principles I think are important. Stay away from systems that require proprietary hardware boards for the software to run. These systems tend to become obsolete quickly or are expensive to upgrade. The less expensive image analysis software programs tend to be easy to use but lack the flexibility and power when confronted with a difficult problem. For overall cost and performance, I think PC based systems are the best. My lab has chosen Optimas software (runs under Win95 or NT) as the main image analysis platform, and sofar it has been able to do everthing we require.
Regards,
John J. Turek, Ph.D. Associate Professor Director, Electron Microscopy Laboratory and Core Laboratory for Image Analysis and Multidimensional Applications (CRISTAL) Department of Basic Medical Sciences 1246 Lynn Hall, G193C Purdue University W. Lafayette, IN 47907-1246 Phone: 765-494-5854 Fax: 765-494-0781 Email: jjt-at-vet.purdue.edu
No, we never did speak regarding Visilog. This may be due to the fact that I do not live in Canada, but even if you are the sales rep for the Northeast, we tried to keep sales people out of the examination, and talk to end users. Only after we had narrowed the choices down to what we thought would meet our needs did we bring the sales people into the picture. I do not believe that in my original memo, I stated anything negative regarding Visilog, but the fact of the matter is, we found that several people considered it not very user friendly (this may have been an older version). With regards to the ALI language not being true C, this is true, but most anything that can be accomplished in C can be done with ALI (maybe it won't be as elegant a program, but it will work). The reality is that ALI is a vector based language and the reason we are doing image analysis is to crunch numbers, and the most efficient way to handle large amounts of numbers is with vectors. As to the rest of your "unique features", it seems that Optimas meets most of them with the possible exceptions of the Matrox Genesis C80 and the price. Oh, by the way, your starting price may be $1000.00, but what does the average system go for?
With regards to your inference that I have an interest in promoting Optimas, I am not on any payroll of any organization that promotes any software, and I do not appreciate your implying that I am. I thought that the purpose of this list serve was for people to give their opinions on a subject when they have one, and that is what I did. I do not think that this is the forum for sales people to make judgments on other people's opinions and I would be interested in hearing what other microscopy scientists and professionals feel about this.
Regards,
David
ln-at-noesisvision.com on 08/01/97 02:11:28 AM
To: David Bell, bruton-at-EMU.UNP.AC.ZA cc: Microscopy-at-sparc5.microscopy.com
The Noesis office is in Canada, just like Matrox, Coreco and Dipix.
But we sell in the United States mainly through a wide number of dealers in every state and region.
Just like thousands of US corporations are located in the US and sell directly to Canada. The opposite also exists.
I can assure you Visilog provides a much wider selection of features than Optimas with our high end version at 6,000.00. Our 3,000.00$ version is similar to Optimas.
Thanks anyway.
At 11:16 AM 8/1/97 -0400, David_Bell-at-millipore.com wrote:
} Dear Luc Nocente,
}
} No, we never did speak regarding Visilog. This may be due to the fact that
} I do not live in Canada, but even if you are the sales rep for the
} Northeast, we tried to keep sales people out of the examination, and talk
} to end users. Only after we had narrowed the choices down to what we
} thought would meet our needs did we bring the sales people into the
} picture. I do not believe that in my original memo, I stated anything
} negative regarding Visilog, but the fact of the matter is, we found that
} several people considered it not very user friendly (this may have been an
} older version). With regards to the ALI language not being true C, this is
} true, but most anything that can be accomplished in C can be done with ALI
} (maybe it won't be as elegant a program, but it will work). The reality is
} that ALI is a vector based language and the reason we are doing image
} analysis is to crunch numbers, and the most efficient way to handle large
} amounts of numbers is with vectors. As to the rest of your "unique
} features", it seems that Optimas meets most of them with the possible
} exceptions of the Matrox Genesis C80 and the price. Oh, by the way, your
} starting price may be $1000.00, but what does the average system go for?
}
} With regards to your inference that I have an interest in promoting
} Optimas, I am not on any payroll of any organization that promotes any
} software, and I do not appreciate your implying that I am. I thought that
} the purpose of this list serve was for people to give their opinions on a
} subject when they have one, and that is what I did. I do not think that
} this is the forum for sales people to make judgments on other people's
} opinions and I would be interested in hearing what other microscopy
Course Announcement: "Optimizing Light Microscopy" When/Where: (a) New York City, November 3,1997 (b) Springfield, MA November 5, 1997 (c) Boston, MA November 7,1997 What: a lively, fast-paced slide lecture and demonstraton for anyone how uses or plans to use a light microscope: students, teachers, medical technologists, clinicians, pathologists, and lab managers. Beginning to more experienced practictioners welcome.
For details... (a) read below (b) send for brochure (c) visit the Microscopy/Microscopy Education booth at MSA - #502
Program: 1. A quick tour around the microscope - getting to know the bits & pieces 2. Koehler illumination & you: 4 critical steps for aligning you and your microscope to reduce headaches, fatigue, and errors 3. Care and cleaning 4. Useful principles for understanding and optimizing imaging 5. Putting the basics to work: a. Troubleshooting b. Understanding Phase and Hoffman Modulation Contrast 6. The Video connection: cameras, computers, and your microscope 7. Bringing out the best: quick, easy, and often free techniques for improving contrast 8. Advanced contrast techniques: Fluorescence and DIC 9. Becoming a better consumer: matching your microscope to your application 10. Questions, Answers, and Information Exchange (Note: Instructor may vary class content slightly to meet the needs of participants)
Free with your tuition: "Optimizing Light Microscopy for Biological and Clinical Laboratories" (Kendall-Hunt, 1997). Nearly 200 pages of helpful hints, quick experiments, and new iedas for getting the best from your microscope. Hot off the presses!
CEU's: 0.6 CEUs, 6 P.A.C.E CEU's Microscopy/Microscopy Education adheres to the guidelines established by the IACET.
Pricing: $150 (includes tuition, breaks, course materials, and copy of book) *****Save $25 if paid by 10/17/97.***** Send three from the same facility and save $50 on tuition for the third person.
Refund policy: Full refund for cancellations made by 10/17/97. After that date, 50% refund or full credit for future class. Substitutions accepted.
Questions: call Barbara Foster or Dr. Ken Piel at MME: (413)746-6931
Registration: Download the form below and fax to (413)746-9311 or call (413)746-6931 and ask for Ken.
Check course you will be attending: ___ New York City, November 3 (#971103) ___ Springfield, November 5 (#971105)* ___ Boston, November 7 (#971107)
Method of Payment: ____ Check enclosed for $ _______________ ____ Visa ____ Mastercard Name on credit card: __________________________________ Credit card number: ___________________________________ Expiration date: ______________________________________ ***If billing address is different from one shown above, please show billing address below: _______________________________________________________ _______________________________________________________ _______________________________________________________
Stephen A. Shaffer wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Inter/Micro 97 Day 4 } } I'll start with some notes on Wednesday's sessions since I neglected to } summarize them last night. I was, er, a bit slow by the time I got } back. :-P The session focus on Wednesday was "History and Art," } although that was only loosely held to. Several of the papers dealt } with artistic subjects but not art conservation per se. } } Gary Laughlin spoke about "A Unique Metallurgical Process From the Early } Bronze Age" in which he described the findings at, and significance of, } a site excavated at the Kestel Mine in the Taurus Mountains of Turkey. } The site dates from the third millenium BC. The examinations indicate } that cassiterite ore was mined and refined at the site to yield a black } magnetic oxide. This would have been more readily separated, due to its } magnetism, than could be accomplished by other means. } } Dr. McCrone gave an "Update on the Turin Shroud" in which he reviewed } several letters he received from Father Rinaldi prior to the Father's } death in 1993. In the letters, Father Rinaldi effectively acknowledged } the proof that the Shroud was painted and actually dated from much later } than the time of Christ's crucifiction, thus was not the true article. } } (For those of you who may not know, Dr. McCrone concluded early on in } the Shroud investigations, and from microscopic observations alone, that } the shroud was a painting. He stood nearly alone in this view and was } vilified for nearly two decades before Carbon dating ultimately } confirmed his conclusions.) } } John Delly gave one of his typical, amazing presentations, this one on } "Hand-colored Microscopical Illustrations." In it John took his } audience on a delightful stroll through 19th century microscopy } publications illuminated with hand-colored illustrations. He } demonstrated the evolution and later de-evolution of the quality of such } illustrations, the variation that one can see from different } illustrators of the same work, and the differences that are seen } edition-to-edition of the same work. Of course, when he became } interested in the subject, John felt compelled to master the art } himself. Through his own study and practice he gained the insignt } necessary to understand and explain the techniques and variations seen } in these historical works. The illustrations are, indeed, beautiful and } many of us are fortunate to own examples of these illustrations in early } works on microscopy. Because of the vast number of color illustrations } necessary to address his subject, it is unlikely that this presentation } will ever be recorded fully in print. (How about a book, John?) Those } of us fortunate enough to be in the room may be the only ones ever to } enjoy this particular product of John's efforts. Thank you, John, once } again. } } Have you ever stood befor an audience wondering why on earth you found } yourself presenting in the particular session where you were? Wayne } Moorehead must have when he spoke on "A Tale of Two Danas; Influences in } Mineralogy" but he soldiered on and did a fine job chronicaling the } lives of two remarkable men. The mineralogists in the audience will } need no introduction to the Danas but I'll just mention for the others } that the elder Dana published his first edition of the System of } Mineralogy in 1835 at the tender age of 24. It was the first such major } scientific work of classification written in English and he and his son } went on to publish or edit a vast array of classic works in Mineralogy. } Together or individually, they edited the prestigious American Journal } of Science continuously for an astounding 95 years, from 1840 to 1935. } One of the most noteable achievements that Wayne mentioned, in my mind } anyway, was when James Dana, in the introduction to a revised edition of } his classic System, abruptly abandoned his entire earlier classification } system as outdated!. Believing that system no longer consistant with } emerging thought, he just as abruptly adopted and described a newer } system which largely stays with us today. I find such honest } self-appraisal and the ability to continue to move forward without } missing stride quite refreshing. } } Wednesday afternoon was occupied by two sessions which would be unusual } at other meetings. Using video microscopy, Anna Teetsov of McCrone } Associates demonstrated some micromanipulation techniques within the } context of creating artistic works on microscope slides by arranging } butterfly scales of various colors into micro-images. Anna and a few } others continue to develop this art form which is particulary unique to } the community of microscopists. One has to have a microscope and } micro-related knowledge and skills in order to produce these beautiful } little creations, then one has to have a microscope to view them as } well. Kind of nice, don't you think? Something we can hold purely for } the aesthetic pleasure and uniquely our own. } } The afternoon was closed with a demonstration by Alan Shin on how one } can construct a working replica of Leeuwenhoek's single lens } microscopes. I did not attend this demonstration as I have on a } previous occasion taken a longer version from Alan in which we got to } actually construct our own microscopes. Comments from those who did } attend and look through the instrument Alan made reflected surprise at } how much one could see and pleasure at the experience of seeing an } insturment of such historical significance actually fabricated. } } Today's sessions were on Forensic Microscopy. Jose Almirall told us } about "Developments in Glass Examination: Automated Microscopy } Techniques and Composition Analysis." Jose's talk was very interesting } and perhaps somewhat troubling to practicing forensic scientists as he } told us of (among other things) a remarkable consistancy in the optical } properties of some glasses, especially window glass manufactured by the } "float" process. The new information for me was the time over which the } products of these plants will remain indistinguishable under } conventional forensic examination techniques. I am not aware of other } time-dependant studies of glass properties but Jose showed data } collected over at least 18 months, during which the product of a float } glass plant was entirely uniform in refractive index. He did however, } offer a remedy for this disturbing finding. He showed that glass } samples which are indistinguishable by refractive index can often be } distinguished by elemental analysis of Fe, Mg, Al, and Zr using } ICP/AES. Now all the forensic people have to do is get themselves one } of these and... ;-) } } Wayne Moorehead gave another excellent paper on Thursday, this one on } "An Introduction to Microscopical Feather Identification." Wayne told } us that the flight and tail feathers of birds are not always useful for } identification but that the down or contour (breast) feathers can be } distinctive, at least down to the order of birds, occasionally to the } family, but virtually never to the genus or species. Still, } identification at this level may prove very useful in a forensic case. } Wayne illustrated how appropriate preparations can be made, what } features of the feather barbule to examine and how they can vary. He } also showed and described the identifying characteristic of numerous } feather types. } } Thom Hopen gave an interesting talk on "Teaching Forensic Microscopy in } Countries Formerly Known as the Soviet Union." Thom responded to a } State Department request that he make numerous trips to various } countries of the former Soviet Union. He has taught courses of fiber } and paint comparison, explosives residue analysis, and basic } microscopy. He found his students to be highly motivated and dedicated } people, anxious for quality instruction in basic forensic microscopy } techniques. Often they are at least adequately equiped though sometimes } have little or no idea how to fully exploit the equipment they have. } (Unfortunately, when it comes to microscopy, this is too often true here } also! My comment, not Thom's.) One can only immagine the difficulty of } teaching in a completely and literally foreign environment, working } through a translator, and using instrumentation previously never seen. } Often, Thom had to set the equipment in proper working order prior to } beginning instruction. But apparently all has worked out for him and } his students and several more trips are planned to continue the } education. } } Well folks, I think I'll stop there. Of course, there were many more } fine presentations and, once again, I'll mention that my choice of } topics covered here in no way reflects badly on the other papers. All } of the presentations were excellent. } } Tomorrow is given over to a tutorial workshop on the Dispersion Staining } technique. It will be given at McCrone Research Institute by Dr. } McCrone and will be attended by twenty-odd students, all that can } reasonably be accomodated in such a hands-on session. For the rest of } us, this afternoon marked the end of another educational, enlightening, } and just plain fun Inter/Micro. } } Special thanks, as usual, to Nancy Daerr who coordinates all } arrangements for these meetings and who, as usual, did an exceptional } job of taking care of us and making our stay wholly enjoyable. } } To all of those interested in these meetings, please note: Next year } marks the Golden Anniversary of Inter/Micro, the fiftieth anniversary. } (Wow!) Plan on attending what promises to be an excellent meeting. } Contact Nancy Daerr for further information, to be put on a mailing } list, etc. She can be reached at McCrone Research Institute, 2820 S. } Michigan Avenue, Chicago, IL, 60616 or simply as ndaerr-at-mcri.org. The } phone numbers at McRI are 312-842-7100 (voice) and 312-842-1078 (fax). } } It's been a pleasure being your ears at Inter/Micro 97. But } tomorrow... Ahhh, Chicago! The architecture, the museums, the Art } Institute! I feel like a nice walk! Happy trails to all, and to all, } Good Night. } } Steve Shaffer } MicroDataware } sshaffer-at-microdataware.comDear Steve,
Many thanks for keeping us all up to date on this valuable meeting. Summaries from one day would have been really nice but summaries from all four were a gift. They were much appreciated.
I am afraid like many many other SEM operators you are using far too high= a kV. If you operate a SEM at 15kV plus you rarely see the true surface of=
the specimen under normal observation conditions. The manufacturers have=
cottoned on to this and improved the lower kV performance tremendously ov= er the last 15 years. You may have noticed how the maximum kV offered was 4= 0 or even 60kV in the early 70's whilst now many offer only 25kV. I rarely=
use a SEM above 10kV unless I am after more sub surface detail when I too=
will use up to 30kV, but only for this reason!
Your problems:-
1. You are using 15kV plus, much much too high with fragile or sensitive specimens. CURE - come down to {7kV 2 to 5 would be best if the sample is really sensitive.
2, You find at lower kV that you simply do not generate enough signa= l to make operation possible at the resolution that you require. =
CURE - move the filament forward in the cathode until you can obtain at least 30uA emission at saturation with the bias set to give th= e highest emission. Without this level of signal sure your task will be ve= ry difficult.
3. The system lacks signal and performance when you lower the kV. CURE - lift the sample in the system because lowering the kV will=
have increased the lens aberrations. Lifting the sample nearer to the le= ns will reduce the aberrations and in doing so the same number of electrons = as you have used at a lower WD will be better packaged giving you a higher current density and a higher signal. How high is high you may ask? Well= I would not dream of operating at {7kV with a WD greater than 10mm, with 3 = to 5 mm being my target depending on the make of instrument. I do not know the instrument you have intimately but if it has a conical lens 3 to 5 mm=
is fine, if it has the old fashioned big flat bottomed lens then 5 to 7mm=
may be better.
4. Resolution is difficult to specify but at 3kV I would expect a correctly set up electron gun to enable you to work with W at 15,000X. I= f Amray offer a low kV anode this would help a great deal. An alternative = is to lift the anode by fitting spacers underneath it! The normal anode to cathode distance is about 1mm for every 2kV. If you are able to lift the=
anode by about 5mm this would make a great deal fo difference to the gun performance. WARNING - PLACE A SIGN ON THE INSTRUMENT "NOT TO BE RUN ABOVE 5kV" whilst you are using it and remove the anode modification prio= r to leaving the instrument.
5. Filament life is going to come into this performance equation. I=
hate people who boast how long their filaments last, I liken this to leaving my car at home whilst I am abroad as I find this to be the most economical use of my car, it doesn't seem to use any fuel at all :-) If you really use a filament it will not last very long but if it makes the impossible possible who cares???
Hope this helps please come back if you need more.
I used to do platelets when I first started in this business. I centrifuged the whole blood (in citrated tube) for a few minutes in a tabletop centrifuge to separate the buffy coat. I then carefully dropped my standard fixative (2+2 glutaraldehyde/paraformaldehyde in 0.1M phosphate buffer) into the tube. After a few hours the buffy coat containing the platelets was lifted out like a pill which could be razor cut in pie-shaped slices. Those were then osmicated, dehydrated, and embedded like any other tissue. Joyce Craig Chicago State University
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Reply to: Unsuscribe
Please unsuscribe me from the list. Thanks!
--------------------------------------
Continuing the discussion of this topic. I used twin jet electropolishing a bit about 1970, but was frustrated by not being able to determine when a shiny surface was produced. (I had been able to see the sample in an old glass system operated manually). About that time, one of the early South Bay 550 polishers was ordered by me here at Argonne because it permitted magnified, IN SITU, viewing of the sample during polishing. This was needed to thin 1 or 2 new materials every week! The ease of use permitted accumulating reproducible data published in a 65 page report, (ANL-80-120), used world wide, a journal cover photo, and 12 other published articles. This work brought me the MSA "Technologist of the Year" award in 1994. About six 550 polishers are used exclusively at Argonne-some as long as 25 years! The unit can do jetting from one side, (for back-thinning to a special surface-lacquer protected), or both sides by inverting the sample after jet polishing about half way thru it. Microshield lacquer, (from South Bay), works great to protect surfaces from etching, dissolves in acetone, and may be thinned to reduce shrinkage when thinning soft, annealled copper for example. Also, the entire 3 m.m. disc surface can be polished via a 3 m.m. jet; using an external "timer/switch",and a D.C. power supply, planar "sectioning" of as little as 100 nm. can be removed from a surface. The jet polishing electrolyte and conditions will work. The large jet can be used to etch a surface for optical photos by simply reducing the "polishing" voltage about 20% for a couple of seconds! The 300 volt, 150 mA. capacity power supply exeeds other manufacturer's units and makes use of non-acid "BK-2" type electrolytes possible-a must for many materials. The line-of-sight optical shut off system can be fitted with a variety of color spectrum light sources for special uses. The standard infra red LED and detector bias may be independently adjusted to give the desired sensitivity setting. It will make electron transparant regions in pure annealled metal such as aluminum-with no hole! Of course the setting is normally set for a 20 micron hole with a very thin edge (quite reproducible, of course). Alignment of the parts is easy and stays set a long time. Even saphire light pipes are available for hydrofluoric acid or bromine/alcohol solutions. PVC plastic parts are available and may be substituted for metal ones for such strong chemical baths. Low temperatures of -50 degrees C. are no problem. The sample is accesible for rapid rinsig after swinging the detent-equipped jet support to one side. In 25 years of use, these instruments have saved one man per year in labor cost, (roughly $100,000/yr.), or $2,500,000--due to the ease and speed with which excellent samples can be made. About 90 to 95% of the samples attempted are good once- conditions are established. In my opinion, all the jet polishers have improved with time, but the South Bay 550 C and 550 D units are unmatched when it comes to working with the newer, difficult materials which should be viewd DURING thinning. They permit me to thin about 300 TEM foils/year in my spare time.
------------------ RFC822 Header Follows ------------------ Received: by qmreceive.anl.gov with ADMIN;16 Jul 1997 16:29:20 +0100 Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by dns2.anl.gov (8.6.11/8.6.11) with ESMTP id QAA27901; Wed, 16 Jul 1997 16:28:09 -0500 Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id NAA04896 for dist-Microscopy; Wed, 16 Jul 1997 13:28:22 -0500 Received: from dns2.anl.gov (dns2.anl.gov [146.139.254.3]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id NAA04893 for {microscopy-at-sparc5.microscopy.com} ; Wed, 16 Jul 1997 13:28:21 -0500 Received: from qmgate.anl.gov (qmgate.anl.gov [146.137.160.48]) by dns2.anl.gov (8.6.11/8.6.11) with SMTP id NAA22774 for {microscopy-at-msa.microscopy.com} ; Wed, 16 Jul 1997 13:32:55 -0500 Message-ID: {n1343062439.13127-at-qmgate.anl.gov}
Does anyone know if we'll be having, and if we should bring, examples of printer output from various printers at the meeting this year? I think it has been an excellent method for quickly comparing the various printers, and since they are ever upgrading the technology and we go right along buying new printers it seems like a very reasonable thing to bring some examples along and lay them out in the computer room again, eh?
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513-529-5712 Fax: 513-529-4243 E-mail: edelmare-at-muohio.edu
"WE ARE MICROSOFT. RESISTANCE IS FUTILE. YOU WILL BE ASSIMILATED."
Nina Stromgren Allen Professor, Department of Botany Box 7612 North Carolina State University Raleigh, NC 27695-7612 Phone: 919-515-8382 (Office), 515-3525 (Lab), 515-2727 (Department Secretary) Fax: 919-515-3436
I see a potential problem brewing here and the hint at tempers going up. It is time to end this particuliar thread in the public forum. This is not the place to carry out long winded commerically related arguments. If you have a problem with this send a private message to me.
COMPUTER INTERACTIVE HIGH-RESOLUTION TRANSMISSION ELECTRON MICROSCOPY
N.C.E.M, LBNL, Berkeley, California
The National Center for Electron Microscopy announces its fourth ANNUAL SUMMER SCHOOL on COMPUTER INTERACTIVE HIGH-RESOLUTION TRANSMISSION ELECTRON MICROSCOPY, including Image Acquisition, Image Processing and Image Simulation, to be held at the National Center for Electron Microscopy during the week of August 25-29, 1997.
The aim of the School is to train participants in the techniques of computer-assisted high-resolution electron microscope image acquisition and image interpretation, including remote-control microscopy. Participants will learn general principles and apply them to specific cases. Participants will be taught the use of computers to obtain images on NCEM microscopes, followed by training in the use of application programs for image interpretation by image processing and image simulation. Participants wanting to apply school techniques to their own projects will be encouraged to extend their visit to NCEM into the next week -- note that this requires a proposal be submitted with advance notice sufficient for project approval.
For more information, please see - http://ncem.lbl.gov/NCEM/workshops.html
:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:. Michael A. O'Keefe, Deputy Head National Center for Electron Microscopy Lawrence Berkeley National Laboratory University of California Berkeley, California 94720 tel: (510) 486-4610 fax: (510) 486-5888 email: maok-at-lbl.gov http://ncem.lbl.gov/ :.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.:.
The July 1997 Microscopy Listserver Archives are now available on line at the MSA WWW Site (http://www.msa.microscopy.com). A healthy month with a file size of 1.3 Mbytes . That according to my records is the most ever posted. In case your curious a total of 1.1 Million Email messages were sent out to the members listserver from this server during the month of July.
Also just a reminder that the Microscopy & Microanalysis 97 meeting is only days away, August 10-14 in Cleveland Ohio. Hotel rooms are at a premium this year so expect a big crowd.
For those of you that can't join us this year, check the MSA WWW pages for updated information on what is happening. Last year we were able to organize a live Internet Video Feed from the meeting. Depending upon the hardware present we will try to organize something along those lines again.
Are you tired of all those opportunities that come along promising that you will get rich if you pay a fee every month to them? Well, the only person getting rich is the one who started it!
I’m just like you. I tried lots of so called money making opportunities. All I ended up doing was adding to my debt!
If your fed up with wasting your hard earned money on someone else's behalf, do yourself a big favor! Visit my Web site and just take a look. I am offering you a FREE Software program that WILL make you money FAST!
Just download it,and check it out!
IT’S FREE!
YOU HAVE NOTHING TO LOOSE!
This is the FIRST, and ONLY time I have ever made any money doing anything off the net!
I started making money in about a week! The money has not stopped pouring in since!!
To visit my Web site and download your copy of this Amazing software, Visit:
Hi, there, I am running a real time captureing for zoospore discahrge of a plant pathogenic fungus through Matrox Inspector, my attempt is to put a clip of this on our web site, the problem I am having now is the file size usually too big. Does anyone know anyway we can resize or edit the avi files into a smaller size? Please drop me your input directly into my mail address. Thanks ahead. KerChung
Can anybody out there supply me with a/the origanal reference on the use of OsO4 as a vapour fixative as for delicate biological material?
ThankyouAlan N Hall Unit for Electron Microscopy Faculty of Biological & Agricultural Sciences University of Pretoria Pretoria 0002 Republic of South Africa Tel: +27-12-420 3297 Fax: +27-12-420 3266
Re: Staining Epon with PAS & IKI. I recently got some ambiguous results wit PAS. We were staining thick (2u), epon-embedded plant sections that had been fixed in aldehyde followed by Osmium, and embedding. We knew that the tissue was lipid rich based on prior work with fresh material and staining of Osmium-treated material with Sudan Black B. The lipid bodies were circular as one would expect. We knew that the tissue could have significant amounts of starch in it, so we used the PAS protocol. Many of the circular bodies which we thought were lipids also stained positively with PAS. I decided to add some aqueous IKI to unstained sections and I was surprised to get a positive reaction which clearly showed the starch grains in amyloplasts. The grains stained brown and could easily be distinguished. The IKI was a potent mix of 1 gm. I & KI in 100 ml, and aged for a year or so. This may be a trivial note (please don't tell me that however), but I thought it might be of some interest.
Question - Might the positive PAS reaction by lipid droplets be due to glycolipids or is this a false positive reaction? I have stained lipid-rich plant material before and have never seen a PAS response like this.
Larry Glitch wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hi everyone! } } I heard - although I can't believe it - that RJ Lee was giving away an } SEM at the MSA meeting. Am I nuts or has anyone else heard about this? } } Thanks! } } Larry
Dear Larry;
YES! it is true RJ Lee Instruments Ltd is giving away (for a 90 day trial period) a Personal SEM at the M&M Conference this year. We are sponsoring a competition asking participants to bring a sample to the RJ Lee booths (400, 402, 404) and take a picture using the PSEM. The best photo will win. Contest rules are available on request. Anyone interested in signing up for some time on the instrument can call Doris Allison (800-573-PSEM) and make an appointment. You can also send an email message and I'll see you get on the calendar. If you are not familiar with the PSEM or Computer Controlled Electron Microscopy, please come see us.
We have received your request to be removed from all of our mailing lists.
To complete the process, please write down the following token:
312411
Return to our website at http://www.iemmc.org/validate.html and input your token and your E-mail address as entered previously. You will then receive an E-mail confirmation of your removal.
i'm currently using spurr's to embedd metal powders (currently Al) with thin oxide layers on the surface (i.e. Al2O3) ...the current protocol is to flat embed the powders on an aluminum weighing dish using spurrs + 1 drop Z60/40 and on the next day...beem capsule embed slices of the flat embedded samples cut with a razor blade...then proceed to microtome for SEM and TEM observation
i'm not sure if the problem is with the sample preparation...should i try a different embedding media?? or if it is with the actual microtoming procedures..
i've managed to get a few good sections but would like more consistency and improvement on the quality of the section...any suggestions would be greatly appreciated....
many thanks in advance
Sincerely, Michael Mandanas Particulate Materials Center Penn State University University Park, PA USA mxm67-at-email.psu.edu
As you can see by now a JUNK Email/Marketing program has discovered the Email address of the Listserver. I have contacted the organization that is running this "service". Please ignore the message that concerns "removal request" you are NOT being removed from the Micrsocopy Listserver. Rather supposedly the Microscopy Listserver is being removed from their system. They unfortunately have a system which sends out the notification to each Email address.
I can't do anything further about it at this time. Let's see how well their "system" works.
We are shopping for a TV rate camera to interface to our Hitachi H-600 TEM via a 35 mm camera port (scope is equipped with STEM). We plan to use this for teaching and when more than one person (i.e. operator and researchers) are working at the scope, not for acquisition of high-quality digital images. To the best of my knowledge these systems work as follows: a small phosphor screen will be moved into the beam path, a camera is focused on this screen and the image is displayed on a small B&W monitor. I have found two vendors so far (Fullam and Gatan), but our purchasing department wants more. If you know of any other vendors for this kind of equipment - or are vendors of it - please reply directly to me by e-mail.
Thanks in advance for any replies.
Heather Owen
Heather A. Owen, Director Electron Microscope Laboratory Department of Biological Sciences University of Wisconsin - Milwaukee (414)229-6816
Please post the following job description for our company on your employment page. Thank you for your help.
Best Regards, Bill Solinski Technical Sales
TECHNICAL SUPPORT AND SALES REPRESENTATIVE
Supplier of high quality instruments and digital imaging products to the domestic & international microscope market, is looking for a motivated person to provide technical & sales support. A BS in biology, medical technology or other sciences is required. We are introducing new products and opportunities exist in this rapidly growing company. Excellent benefits include choice of fully paid PPO or HMO medical plans, dental, prescription, vision, life ins., 401K and educational reimbursement.
Send resume to: Diagnostic Instruments, Inc.
Attn: Sales Department
6540 Burroughs
Sterling Heights, MI 48314 e-mail: info-at-diaginc.com
Best Regards, Bill Solinski
Diagnostic Instruments 6540 Burroughs Sterling Heights, MI 48314 Phone: 810-731-6000 FAX: 810-731-6469 Website: www.diaginc.com e-mail: info-at-diaginc.com
--IMA.Boundary.192357968 Content-Type: text/plain; charset=US-ASCII Content-Transfer-Encoding: 7bit Content-Description: cc:Mail note part
Yes we are putting our images on CD ROMs in various Photoshop formats and reading them on both Power Mac/Macs and PCs - no problem. We haven't gone to the multiple session, do it yourself CDs but we have someone here make them on a higher quality, dedicated system, presumably because it is more reliable in giving us an error free disk. Also I disagree about Zip drives, we use them to transfer large numbers of images from our lab to our customers; however, I've been told by computer sales that Zip drives are discontinued. Any confirmation? If so, that goes along with a previous message about archiving digital images when the hardware won't be around to read the data. I have already run into this with data stored on 8 1/2" floppies for a Kevex EDXS instrument that is no longer around. Here is a business opportunity for someone who wants to archive all this old equipment to read archived data; of course, you'll need extra units for spare parts....:-)
Hope this answers your question, Tom.
Damian Neuberger neuberd-at-baxter.com
Our multi-user facility is currently archiving our confocal and LM digital images on Panasonic optical disks (re-writable, very stable -at- about $125 for 1 GB). The disadvantage is that few of our users have their own Panasonic drives so most people simply archive the images at our core and then move the ones they want by FTP as needed. I would like to switch to a more universal medium - namely CD ROM's. My understanding is that CD's can now be written to in multiple sessions so you don't need to fill an entire disk at once. Furthermore, it is my understanding that a disk of TIFF images should be readable by both IBM/WINTEL and Mac/PowerPC types computers. Is anybody actually doing this? Comments on how reliable are the recorders, which ones are best, pitfalls, etc would be appreciated. Before I get a dozen advocates of ZIP/Jazz drives, I don't want to go that route since that they are not as ubiquitous as CD drives. Thanks in advance.
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
Michael P. Mandanas and microscopists: You are not describing the actual problem, but I expect that the Al particles pull out of the sections. Use the hardest mixture of Spurr's and over-cure a bit. Use a small block-face and a diamond knife, best one with a more obtuse angle than the biology knives have. For SEM specimens could be more effectively ground. Do not use carborundum powder because particles will be embedded within your specimen. Diamond paste would be a lesser problem but best are diamond lapping films (from EMS or ProSciTech), which are plastic films with embedded, not glued diamond particles. Regards Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 400+ Links, MSDS ************************ http://www.proscitech.com.au
} i'm currently using spurr's to embedd metal powders (currently Al) with } thin oxide layers on the surface (i.e. Al2O3) ...the current protocol is to } flat embed the powders on an aluminum weighing dish using spurrs + 1 drop } Z60/40 and on the next day...beem capsule embed slices of the flat embedded } samples cut with a razor blade...then proceed to microtome for SEM and TEM } observation } } i'm not sure if the problem is with the sample preparation...should i try a } different embedding media?? or if it is with the actual microtoming } procedures.. } } i've managed to get a few good sections but would like more consistency and } improvement on the quality of the section...any suggestions would be } greatly appreciated.... } } many thanks in advance } } Sincerely, } Michael Mandanas } Particulate Materials Center } Penn State University } University Park, PA USA } mxm67-at-email.psu.edu } }
I am working with decalcified bone which has been fixed with osmium and embedded in Spurrs for TEM. There are sections being taken for LM which we would like to do optical staining on but I am encountering alot of difficulty.This is not my field of study, I am a work-study student and would appreciate any help/suggestions for the following:
Semi-thin and ultra thin section of decalcified bone (14% EDTA) which is fixed in osmium and embedded in Spurrs. I have tried a 2%NaOH/absolute alcohol to deplasticize then H2O2 to remove the osmium and have been working with Villnueva trichrome bone stain and tetratchrome bone stain. The specimens are only picking up a very small amount of stain after 24 hrs of staining and they are degraded (probably from the H2O2).
If anyone has further suggestions please let me know. Thanks.
i apologize for forgeting to state the actual problem...i guess things done in haste does go to waste. anyway, what i forgot to mention in my previous email is that the particles seem to pullout and the spurrs does not hold the particles strong enough during sectioning-there are void spaces around the particles and the surface is rough...another problem we've encountered is charging on the TEM when attempting to get higher magnification images - a spurrs stability under electron beam problem? i hope this clarifies my problem and again, i apologize for the confusion
sincerely
Michael Mandanas Particulate Materials Center Penn State University University Park, PA USA mxm67-at-email.psu.edu
On July 31 Ian Laren asked: } Also, does anyone have any idea why oily blobs (obviously from oil in the } vacuum system) sometimes appear on the sample in the very place that you } are observing (as opposed to any other place on the sample)? } } Thanks }
Ian and all:
The probable mechanism is that oil vapor molecules are ionized by the electron beam (Mass spectrometers use this ionization method), and then the positive ions are attracted to the negative charge deposited by the beam on the sample. Thus a hydrocarbon polymer is deposited exactly on your area of interest.
A long discussion of contamination and its control appears at our web site.
Ronald Vane XEI Scientific SEM-CLEAN anti-contamination systems (650) 369-0133 http://www.msa.microscopy.com/SM/XEI/XEIHomePage.html
Recently, I attempted to obtain 70 nm sections from a specimen embedded in Spurr's. The plastic was separating from the tissue shortly after the cut. Furthermore, these sections are not holding up to the beam. I believe my problem is an incomplete infiltration. I allowed the blocks to sit in a 90 degrees C oven over the weekend to see if that would help. The problem, however, continues to exist. Is anyone familiar with any methods of depolymerization and reinfiltration of embedded specimens? Any other suggestions that may help would be greatly appreciated.
I prferred Epon to Spurrs because the hardness of the Epon can be adjusted readily by different ratio's of mixtures A and B as described in the Biology handbooks. Look into any biology related microscopy book to get the proper ratios. I think you will find that the epon is much harder. I don't know if that will solve your problem. As for the charging, try a thin coating of carbon before inserting into the TEM.
Good luck.
Roberto R. Garcia EMF Manager Wright State University
Michael Mandanas wrote: ======================================= i'm currently using spurr's to embedd metal powders (currently Al) with thin oxide layers on the surface (i.e. Al2O3) ...the current protocol is to flat embed the powders on an aluminum weighing dish using spurrs + 1 drop Z60/40 and on the next day...beem capsule embed slices of the flat embedded samples cut with a razor blade...then proceed to microtome for SEM and TEM observation
i'm not sure if the problem is with the sample preparation...should i try a different embedding media?? or if it is with the actual microtoming procedures.. ================================================= Our laboratory has been diamond knife thin sectioning metal powders, including those of aluminum for over twenty seven years. I would like to summarize our experiences, some of which actually led to some new product concepts (e.g. the concept of a materials science diamond knife):
1] Aluminum is a special case because relative to most other powers one might want to section, it is very soft. If it further "special" because if there is indeed an oxide layer, e.g. like a layer of anodization, since the adhesion of the oxide layer is not the greatest, one has to use vacuum embedding. Otherwise, the "pores" of the oxide layer do not get impregnated with the resin and there is then an extraordinary level of "pullout" of the particles.
2] After trying all of the various embedding resins, we have settled on our own SPI-Pon(TM) resin kit. We suspect, but don't know it for a fact, that at least some of the so-called "Epon(TM) 812 replacement" kits offered by others would work just as well. It seems like one has not only the maximum ease with which the hardness can be controlled, but for the hardest of powders, it seems, at least to us, that this resin system permits the curing of the hardest possible block, yet when sectioning, there is less of a tendency to "chatter" and "compress".
3] Diamond knives must be used in this kind of application, and trying to use glass will just turn out to be a grand exercise in frustration. And further on that, we would certainly want to be using a "materials science" diamond knife. Now at the risk of sounding "commercial" I would like to say that there are three schools of thought on the matter of "materials science" diamond knives: a) The concept itself is a gimmick, b) make your knife with an angle that is much more blunt (e.g. not such a sharp angle), or c) finish off the knife with the same angle as is used for life science knives generally (e.g. 45 deg.), but perhaps not worry about the last of the fine striations, since any that are there are going to be small indeed compared to the larger ones that will be put in on the first slice.
We ourselves are part of the last school of thought on this issue. It is certainly not a gimmick because why would anyone in their right mind pay top dollar for the perfect edge, only to reduce it to a state that is no better, and probably worse than a "materials science" knife from that third school when it is brand new? The concept of using a more blunt of an angle on the knife, while in theory (and usually in practice) such an edge will last longer, because of the less "sharp" edge, compression effects tend to be greater as well as also, the instances of particle "pull out". Sometimes these effects are sufficiently profound that usable sections can not be made at all (it depends on the powder being sectioned). And in the end, if usable sections are obtained, usually more sections have to be made, putting more wear and tare on the knife edge, so that in the end, its useful lifetime is not all that much longer.
4] We have found the above to be true, not just with the relatively spherical metal powders, but also with aluminum flake, of the type used in automotive (metallic) paint coatings.
While there is no question there is some "art" (as well as experience) involved here, once the "secret" is known, it is no longer magic. On the other hand, I have suspicions that there are some who have obtained quite excellent sections using other embedding resins and perhaps diamond knives from the second school as well, so even we won't claim to know all of the "magic". I can, however, speak only from our own experiences.
Disclaimer: Our firm offers diamond knife thin sectioning,on these kinds of materials, as a service, and for a fee for others and we also offer the SPI Materials Science line of diamond knives for persons wanting to do this type of work themselves. Information about these products and services can be found at the website address given below.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
At 08:27 PM 8/4/97 -0400, Bill wrote: } I have downloaded your database files but cannot unzip xray_ms.zip, it gets } a crc error when I try. I downloaded xray.xls and xray_ms.xls but neither } would open with Excel 7.0 on a PC. Can you offer any suggestions? } I'm getting similar feedback from others ... and I've just tried to open the files without luck (... weird ...). I've just opened the working spreadsheet (Excel v.7). It actually is 2 sheets ... one of which is the database and the other sine-theta values for Cameca spectrometers, which can be easily be converted if you change the cystal 2D and k values, and the sine-theta range. It could also be easily modified for LiF and wavelength geared spectrometers (Note: the sine-theta sheet has hidden columns and frozen rows for visibility). I will zip it then unzip it for a check, and let you and others know I can dump the separate sheets as other types of files or text files. The zipped Excel (v.7) spreadsheet xrayxls7.xls (xray_ms.zip) will be available via anonymous FTP at whitewater.uoregon.edu/share/cameca/ ... the other types of files can be requested.
cheerios, shAf
BTW ... the 2Mb zipped file tested, unzipped and loaded A-OK ... the XLS file also exists at the FTP site but it is 6Mb. Let me know if there are any problems ... I've also been successful in creating a PDF file but I'm waiting to get the 2D and k values for one more xtal before it is finished.
{\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/epmahome/
The Iowa Microscopy Society meeting is to be held on September 25th and 26th, 1997. An immunocytochemistry workshop is being offered, and advance registration is suggested. More information on the workshop and the Iowa Microscopy Society's meeting can be found at http://www.uiowa.edu/~cemrf/cemrf/ims_announce.html or you can give Kenneth Moore or myself a call at 319-335-8142.
Randy Nessler rnessler-at-emiris.iaf.uiowa.edu Views expressed are my own.
recently i have been studying polymer distribution with ceramic powders (in this case Al2O3)...in one case, i stained polyvinyl alcohol with RuO4, and there is a gradient of the polymer distribution in the powder...now my question is, what would be the best way to determine and/or calculate the concentration gradient / distribution of the polyvinyl alcohol...does the RuO4 concentration directly translate to polymer concentration? ..if so, can i just take sections and do mass spec each of the sections?? should i be working with a different staining agent for PVA, what about other polymers like polyethylene glycol, acrylics ?? a more general question would be, what is a recommended refernce for polymer staining..i've read Linda Sawyer's "Polymer Microscopy" textbook but i was looking for a more detailed discussion...if mechanisms are included, the more help it would be... any suggestions would be greatly appreciated...
thanks again in advance sincerely
Michael Mandanas Particulate Materials Center Pennsylvania State University University Park, PA 16801 USA mxm67-at-email.psu.edu
Updates for both Mac and Windows versions are now ready for downloading at http://Members.AOL.com/ImagProcTK/update.htm
Intensity Profile is now available for the PC, Autocontrast for both platforms expands contrast over the full range of the image (great for images with shading, or TEM sections with varying thickness), and there is a spreadsheet with examples for data analysis (statistics, graphs, and sphere unfolding).
DIRECTIONS FROM CLEVELANE HOPKINS INTERNATIONAL AIRPORT TO DOWNTOWN CLEVELAND VIA THE RAPID TRANSIT AUTHORITY (RTP-RAPID)
From the baggage claim level at the airport, proceed down one level via the escalators in the center of the baggage claim. Continue following signs to the rapid transit station. Exact change is required - $1.50. Take the rapid transit to the Tower City - Downtown Terminal which is the last stop. Proceed through the turn styles.
If your accommodations are at the Ritz Carlton, once through the turn styles, turn right and proceed up the two sets of short escalators. The entrance to the Ritz Carlton is on your left.
If your accommodations are at the Renaissance Cleveland Hotel, once through the turn styles, proceed left up to the long set of escalators. At the top, make an immediate left and then another immediate right and follow the signs towards Public Square. Once you reach the Disney store, turn left and continue straight through the double set of glass doors. Proceed up the staircase and you are in the lobby of the Renaissance Hotel.
If you are staying in another downtown property, follow the directions as listed above for the Renaissance Hotel however, at the Disney store, proceed straight through the glass doors heading outside where you will find taxi's.
The Marriott and Sheraton are 2-1/2 and 4 blocks respectfully if you choose to walk. Once outside walk across Public Square towards the Key Bank Center where the Marriott is located. The Sheraton is within sight from the Marriott entrance, next to the Cleveland Convention Center.
{P} {B} DIRECTIONS FROM CLEVELANE HOPKINS INTERNATIONAL AIRPORT {/B} {BR} {B} TO DOWNTOWN CLEVELAND VIA THE RAPID TRANSIT AUTHORITY {/B} {BR} {B} &n bsp; (RTP-RAPID) {/B}
{P} From the baggage claim level at the airport, proceed down one level via the escalators in the {BR} center of the baggage claim. Continue following signs to the rapid transit station. Exact change is {BR} required - $1.50. Take the rapid transit to the Tower City - Downtown Terminal which is the {BR} last stop. Proceed through the turn styles.
{P} If your accommodations are at the Ritz Carlton, once through the turn styles, turn right and {BR} proceed up the two sets of short escalators. The entrance to the Ritz Carlton is on your left.
{P} If your accommodations are at the Renaissance Cleveland Hotel, once through the turn styles, {BR} proceed left up to the long set of escalators. At the top, make an immediate left and then another {BR} immediate right and follow the signs towards Public Square. Once you reach the Disney store, {BR} turn left and continue straight through the double set of glass doors. Proceed up the staircase and {BR} you are in the lobby of the Renaissance Hotel.
{P} If you are staying in another downtown property, follow the directions as listed above for the {BR} Renaissance Hotel however, at the Disney store, proceed straight through the glass doors heading {BR} outside where you will find taxi's.
{P} The Marriott and Sheraton are 2-1/2 and 4 blocks respectfully if you choose to walk. Once outside {BR} walk across Public Square towards the Key Bank Center where the Marriott is located. The {BR} Sheraton is within sight from the Marriott entrance, next to the Cleveland Convention Center.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello all, } } Recently, I attempted to obtain 70 nm sections from a specimen embedded in } Spurr's. The plastic was separating from the tissue shortly after the cut. } Furthermore, these sections are not holding up to the beam. I believe my } problem is an incomplete infiltration. I allowed the blocks to sit in a 90 } degrees C oven over the weekend to see if that would help. The problem, } however, continues to exist. Is anyone familiar with any methods of } depolymerization and reinfiltration of embedded specimens? Any other } suggestions that may help would be greatly appreciated. } } Sincerely, } } Dan Caruso c/o Eugene Gordon } } Your best bet would be to start over. However I was faced with a similar problem two years ago with some tissue which had been very poorly infiltrated (in another lab)and brought to us for evaluation. You need patience: 1. Cut the tissue out of the existing blocks. Trim as closely as possible. 2. Expose the tissue to numerous changes of propylene oxide in vials which are on a rotator. You will see the unpolymerized resin gradually enter the PO. Keep changing fluids. This may take 48 hours. Tissue will get softened. When you feel that no more changes are forthcoming, reembed the tissue in the same protocol as the first, making sure that each time the tissues remain in motion. Lengthen the infiltrations steps, especially the first one which is likely to be mixture of resin and intermediate agent. Reembed and polymerize and hope. The original polymerization may have gone far enough so that the above method is not successful. The whole procedure may take a week, but if the redoing is successful you may save valuable tissue. If the method is not adequate for sectioning, you must start over. If the sections are "delicate" pick them up on formvar coated grids and carbon coat them. 3. What was the cause of this problem? It may have been water! Was propylene oxide or acetone, or alcohol left in the tissue? Or was it truly only inadequate infiltration??? It is important to make a distinction. Good luck. Hildy
Email: hyphae-at-msn.com Name: Scott Mcphee School: Santa Rosa Junior College, California
Greetings,
I am a beginning Mycology and Phytopathology student and am looking for a good quality inexpensive ( {$400) microscope. From the prices I have seen while browsing various sites it looks like this will probably be used due my limited funds.
I would like to be able to spore and cellular characteristics while I am at home. I can take a couple items to school and look at them there, but I often have large amounts of things to study and it would be much more conveniant to be able to do this at home. The fungi often turn to mush also before I can get them to the lab at school.
Could you reccomend a power range and perhaps a model?
I was given a block of paraffin wax with specimen embedded in it. I would like to know how can I get away the wax to get out the specimen. It is rather urgent and I would appreciate immediate reply if possible. Thank you very much.
Many people have problems obtaining information easily from their TEM and=
SEM, I see it almost every day. The area of instrument set up that causes=
more of these problems than any other is that of filament position.
Most people set the filament a long way from the cathode aperture and thi= s leads to a long filament life but a low emission current. Moving the filament forward increases emission current but decreases filament life. =
With care and the additional use of the bias or emission control this forward movement of the filament will result in an improvement in performance - resolution. One problem, people place more credence in having a long filament life than in getting more from their microscope! =
Many buy a new machine because they feel the reason they cannot obtain th= e result they want is down to the microscope. Forget filament life if you want more from your machine, in my experience an instrument will be transformed if you push the gun a little harder. I could tell you so man= y stories where we have done this, much to the amazement of the owner and delight of the dissatisfied customer!
As for changing filaments the more you do it the less frightening it becomes, its really not difficult and to be honest if you want more from your microscope the cost is very little.
Why should people calibrate their microscope you may ask? If you do not take what is a pretty simple step how do you know that the instrument is working correctly? Is it not better to test the microscope via resolutio= n, contamination and calibration than to have a hoard of customers banging o= n your door complaining that their results are poor; too late! Preventativ= e maintenance, spotting problems at higher levels of operation than the normal in your laboratory, should be part of every well run laboratories routine. Spot the problems before they become a disaster and get them fixed before anyone else notices, thats they way to run a unit! It is n= o good saying the engineer does it one or twice a year, (does he really?) that gives a long period of uncertain operation and possible problems. I= n addition, if you learn to operate your microscope at higher levels than i= s the norm you improve your own techniques and powers of observation.
On one of my hobby horses, I believe that every laboratory should test it= s instruments and its operators routinely to see how they all perform. If you set a standard where you are testing in this way you instantly raise the levels of expertise and have standards which may be used over many months to prove that the laboratory is moving forward. Without such a regime laboratories stagnate! Everyone feels they do a great job but wit= h no form of standard how do they know? We talk about Quality in electron microscopy in relation to how we set analytical standards and calibration= ; good! But an even more important question which is totally ignored is "how good are the operators?". I have said before microscopists are supposed to be scientists, real scientists would constantly test themselves!
Many people have problems obtaining information easily from their TEM and=
SEM, I see it almost every day. The area of instrument set up that causes=
more of these problems than any other is that of filament position.
Most people set the filament a long way from the cathode aperture and thi= s leads to a long filament life but a low emission current. Moving the filament forward increases emission current but decreases filament life. =
With care and the additional use of the bias or emission control this forward movement of the filament will result in an improvement in performance - resolution. One problem, people place more credence in having a long filament life than in getting more from their microscope! =
Many buy a new machine because they feel the reason they cannot obtain th= e result they want is down to the microscope. Forget filament life if you want more from your machine, in my experience an instrument will be transformed if you push the gun a little harder. I could tell you so man= y stories where we have done this, much to the amazement of the owner and delight of the dissatisfied customer!
As for changing filaments the more you do it the less frightening it becomes, its really not difficult and to be honest if you want more from your microscope the cost is very little.
Why should people calibrate their microscope you may ask? If you do not take what is a pretty simple step how do you know that the instrument is working correctly? Is it not better to test the microscope via resolutio= n, contamination and calibration than to have a hoard of customers banging o= n your door complaining that their results are poor; too late! Preventativ= e maintenance, spotting problems at higher levels of operation than the normal in your laboratory, should be part of every well run laboratories routine. Spot the problems before they become a disaster and get them fixed before anyone else notices, thats they way to run a unit! It is n= o good saying the engineer does it one or twice a year, (does he really?) that gives a long period of uncertain operation and possible problems. I= n addition, if you learn to operate your microscope at higher levels than i= s the norm you improve your own techniques and powers of observation.
On one of my hobby horses, I believe that every laboratory should test it= s instruments and its operators routinely to see how they all perform. If you set a standard where you are testing in this way you instantly raise the levels of expertise and have standards which may be used over many months to prove that the laboratory is moving forward. Without such a regime laboratories stagnate! Everyone feels they do a great job but wit= h no form of standard how do they know? We talk about Quality in electron microscopy in relation to how we set analytical standards and calibration= ; good! But an even more important question which is totally ignored is "how good are the operators?". I have said before microscopists are supposed to be scientists, real scientists would constantly test themselves!
} I was given a block of paraffin wax with specimen embedded in it. I } would like to know how can I get away the wax to get out the specimen. } It is rather urgent and I would appreciate immediate reply if possible. } Thank you very much.
Would your specimen be harmed if you used solvent to remove the paraffin? Hexane, heptane or Petroleum Spirits 60-80, warmed slightly if necessary, would be suitable. They are not very toxic (but don't breathe too much), but they are highly flammable.
Several washings in a small quantity of solvent each time are MUCH MORE EFFICIENT than washing in one big lot of solvent (Bunsen's dilution law).
+------------------------------------------------------------------------+ | 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 | +------------------------------------------------------------------------+
IF the speciment can take xylene, and wasn't directly embedded in paraffin, then:
Cut the specimen out of the wax, trim away as much wax as possible, then soak in xylene or a xylene replacement (like HistoClear). Make several changes to make sure you get all of the wax out. Times depend on specimen size and nature. 3 X 10 minutes might work, or might be too short. Look for a transparent quality to the specimen, that should indicate that all the wax is gone and completely replaced with xylene.
What you do next depends on what you're after. The specimen can be backed down through a xylene-alcohol series to 100% alcohol, and then through alcolhols to 70% EtOH, or even to water. Just reverse the usual schedules using in LM paraffin staining.
Whether the specimen can go through this and stay in good condition depends on the specimen, and to some extent on how much you need fine details. Most can, but not all (I don't have any examples in the top of my head).
However, before you de-embed, what do you what to look at? If the specimen is say, a whole insect, you can trim away the excess wax, and then, while still embedded, "dissect" the specimen by carving away the unwanted bits. The wax holds the specimen together, and you can carve on the curve, instead of just flat planes like a microtome does. (Also, the microscope lights [use fiber optics] melt the wax locally, and the streams of molten wax wash away the debris.) After carving, then proceed through the series as above.
Phil } } } I was given a block of paraffin wax with specimen embedded in it. I } } would like to know how can I get away the wax to get out the specimen. } } It is rather urgent and I would appreciate immediate reply if possible. } } Thank you very much. } } } } } } } } Catherine Tang }
}}}}}}}}}}}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{{{{{{(((( Philip Oshel Station A PO Box 5037 Champaign, IL 61825-5037 (217) 355-1143 oshel-at-ux1.cso.uiuc.edu *** looking for a job again *****
What do you want to do with the specimen? Two quick ways to remove paraffin- 1) Melt it away in a 65 degree oven and 2) Dissolve it away with xylene. More info is needed to answer your question.
Ed Calomeni Dept Pathology Medical College of Ohio Toledo, OH 43699 emlab-at-opus.mco.edu
We deparaffinize with xylene. The time required will depend on the size of the tissue, so I suggest you cut out a small piece if possible. We then rehydrate step wise to water. Then we handle it as a normal TEM specimen. } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } At 02:10 PM 8/6/97 +0800, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America Scientific Director, ICBR Electron Microscopy Core Lab PO Box 118525 Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
The most straight forward way to remove the paraffin would be to trim the block as close to the tissue as possible and place it in xylene or histoclear to dissolve the rest of the paraffin away. I would do at least three changes, the time of the change would depend on the block size, then rinse out the paraffin/xylene with 3 changes of 100% ethanol and process in your new media, starting from the 100% ethanol step. I have done this with some success, though often the tissue that's processed for LM doesn't look so hot at EM. That depends on how it was fixed.
Good luck,
Karen Pawlowski PhD student/Histology Tech. UT Dallas/ UT Southwestern Medical Center Dallas
On Wed, 6 Aug 1997, Tang Ee Koon wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I was given a block of paraffin wax with specimen embedded in it. I } would like to know how can I get away the wax to get out the specimen. } It is rather urgent and I would appreciate immediate reply if possible. } Thank you very much. } } } } Catherine Tang }
Hello everyone, I am looking for a list of vendors of used light microscopes and parts. I would like to have their phone number or other information that will help me contact them. You may reply directly to me. Thank you Sincerely Karpura kkommine-at-mbl.edu Marine Biological Laboratory Woods Hole
We have been using calibration tolerances of + or - 5% for both our magnification and spectrometer calibrations for quite some time now. Apparently these values have been established through supplier and user inputs quite some time ago. I am curious as to whether these values are consistent with those currently used or whether they are too loose. We are ISO 9002 registered and quality measures are very much a concern. TIA
} I am a beginning Mycology and Phytopathology student and am looking for a } good quality inexpensive ( {$400) microscope. From the prices I have seen } while browsing various sites it looks like this will probably be used due } my limited funds. } } I would like to be able to spore and cellular characteristics while I am at } home. I can take a couple items to school and look at them there, but I } often have large amounts of things to study and it would be much more } conveniant to be able to do this at home. The fungi often turn to mush also } before I can get them to the lab at school. } } Could you reccomend a power range and perhaps a model?
You'll be able to get by with 10 & 40x objectives, but 100x (oil immersion) would be nice, and maybe 4x for the big stuff. 10x eyepiece. A condenser is highly desirable. Try the "surplus" departments at U.C.S.F. and U.C. Berkeley for some really good buys. Take a friend who knows scopes with you if possible. Ask your college to buy the U. of Washington CD-ROM listed in the MICRO bibliography (address below).
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
Tang wrote regarding retrieval of specimen from wax} Simply cut down the wax to the size of the specimen and place in several changes of xylene (could be warmed slightly in a hood). Assuming you wish to attempt TEM microscopy on the specimen at this point once all the wax is removed you don't need to dehydrtate since the specimen is already dehydrated and attempts to use osmium tetroxide seem futile. Next process in propylene oxide and infiltrate with plastic as usual and stain the heck out of the grids. Results are usually very poor to marginal but diagnoses have been made on some tumors this way.
fhayes-at-dow.com wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Does anyone know of a vendor that supplies lacey carbon coated Be grids? } } If not, are there any suggestions out there regarding how to make such a } substrate? } } Fred Hayes } The Dow Chemical Co } Analytical Sciences Laboratory } 1897 Bldg., E78 } Midland, MI 48667 } 517-638-2203 } 517-638-6443 fax }
Dear Fred, Ladd Research has done lacey carbon coated Be grids in the past for customers. Please contact us via e-mail or 1-800-451-3406 and we can discuss pricing and what size mesh you would like it done on.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Clean UA/lead stain 8/6/97 12:49 PM
Has anyone heard of using a few drops triton-X detergent in aqueous uranyl acetate to help keep stain clean when staining with uranyl acetate and lead? If so, does anyone have a logical explanation why this may work? Thanks in advance for the help. Linda Chicoine Center for Cell Imaging Yale University New Haven, CT
I've been ignoring the microscopy meeting that is coming to Cleveland and suddenly my boss said to me today, "hey, we should think about going to this thing next week". We work in Cleveland so thats not a problem. Could someone send me information like schedules and fees and seminars and classes, etc, etc, etc. Thanks. Mark Darus
Intel Corporation currently has an open position for a SIMS technician at its Santa Clara site's Materials Technology department. The SIMS group supports the Santa Clara site's development and fabrication facilities that produces state of the art microprocessors. The Materials Technology department's scope encompasses the whole process from silicon to a packaged device that includes process trouble-shooting, transfer and equipment qualifications, device failure analysis/fault isolation and new materials development to name a few. The department has a wide variety of state of the art instrumentation, including: SIMS, Auger, TEM, SEM/EDS, FIB, AFM, XRF, ICP-MS, FTIR, Raman, Pico and Nano Indentation.
Job Description:
The technician opening involves second shift operation of Quadrupole (Atomica) and Magnetic Sector (Cameca IMS- 6F) SIMS tools. SIMS is an Ultra High Vacuum analytical tool which uses ion beam sputtering combined with mass spectrometry detection to analyze dopant and contamination levels and distributions in patterned and unpatterned wafers. Duties will include data collection, data processing, report writing, interfacing with other engineers and customers, communicating results, and workflow duties to keep the lab running smoothly. The successful applicant must be self-motivated and capable of working with minimal supervision.
Qualification: An AA or BS degree with Engineering or physical science background or equivalent experience is required. Ability to work in a team oriented environment and good communication skills are critical. Experience with analytical equipment or Ultra High Vacuum or Vacuum systems is desired. Experience with SIMS and knowledge of Semiconductor processing methods is a definite plus.
Intel's industry leading total compensation package includes a competitive salary, stock options, annual employee bonus plan, bi-annual employee cash bonus payout, periodic paid sabbatical leave, and retirement plan. Intel is an equal opportunity employer.
Send resumes to or for more information contact:
Gabi Neubauer or Jerry Hunter Intel Corporation 2200 Mission College Blvd., M/S: SC2-24 Santa Clara, CA 95052
Phone: (408) 765-2241 or 765-2316 FAX: (408) 765-2393
Peter Tarquinio Evex Analytical 857 State road Princeton, NJ 08540
----------
I've been ignoring the microscopy meeting that is coming to Cleveland and suddenly my boss said to me today, "hey, we should think about going to this thing next week". We work in Cleveland so thats not a problem. Could someone send me information like schedules and fees and seminars and classes, etc, etc, etc. Thanks. Mark Darus
One of the major problems with crystals and crystal lattice specimens as = a TEM test specimen is the need to determine the level of astigmatism in th= e image. As a novice service engineer needing a crystal lattice resolution=
picture you soon learn to place a little astigmatism in the image and hop= e! Its much easier than trying to find why you cannot resolve the lattice.
The best TEM test specimen to cover a wide range of operator levels is th= e holey carbon film. Tom Mulvey stated that "the minimum discernable fring= e level is similar to the point to point resolution of the instrument"
A holey carbon film at 200,000X is a test for anyone, the object being to=
obtain the finest fringe that is even all round the hole. It tests the operators skills and their ability to truly observe an image. A through focal series is required but it is a waste of time doing this within two hours of switching on the kV as this will take time to stabilse. I have discussed heat gained in the tank needing to equal heat lost in order to obtain stability within the past few months on e-mail.
Correct the astigmatism at double the photo mag and then at the photo mag=
set the focus so that you just see an over or under focus fringe. Take a set of pictures through focus and then measure the centre of the black fringe to the centre of the white fringe ON THE NEGATIVE but only if the fringe is even. Fringes tell you a good deal about the instrument and its=
alignment (Reference Monitoring & Maintaining the TEM) Most people start=
off with about a 1 to 1.5nm resolution and if you are really good you may=
attain 0.45nm at which point the carbon structure starts to interfere wit= h the fringe.
What do you learn. =
1) You will see if the high voltage is stable - it is not perfect if you=
get focus drift, if you do not see some focus drift I would be surprised.=
2) You will notice specimen drift when you first insert the rod 3) You will realise the importance of a good anticontamination device as the hole shrinks in size. 4) You will probably need more current - its that filament position again= ! 5) How difficult it can be at this level to correct astigmatism, you are looking for a fringe like a piece of cotton not a ships hauser. 6) Typical settings - 20-25uA emission, 0.5 to 1micron spot size, CII overfocus, eucentric point, 4 second exposure to test the machine (0.5 second exposures test nothing). =
7) I do the test without an objective aperture so as to test the microscope, not the aperture cleanlyness.
Hope this helps please come back if you need more information.
Since my days as a service engineer it seems to have been accepted that o= ne could run a TEM within plus or minus 5% of the mag readout. You usually find that if the magnification is out across the range it is due to the high voltage level, specimen not at the eucentric point, or the final len= s level. If the calibration is only out after a certain point it suggest that the lens which switches in at this point is at fault.
On the SEM unless you work with perfectly flat specimens magnification accuracy is a bit of a joke! Again it seems that if you check as many instruments as I see each year that 95% are within plus or minus 10%, and=
within plus or minus 5% X to Y. Either way its usually an engineer job t= o tune the scan circuits to improve the performance.
Hello Catherine and all: Place the block in xylene and give it gentle agitation. Change the solvent a couple of times. Unless the block is very large you can de-wax the block overnight. Then dehydrate, opposite to hydration steps, but more rapidly. For the last step use single strength buffer and then fix the specimen in OsO4 and process as normal for EM specimens. Unfortunately specimens initially fixed for histology are never as good as those that were initially prepared for EM, in fact they are disappointing. However, they may show what is required and that is what matters most. Regards Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 400+ Links, MSDS ************************ http://www.proscitech.com.au
} } I was given a block of paraffin wax with specimen embedded in it. I } would like to know how can I get away the wax to get out the specimen. } It is rather urgent and I would appreciate immediate reply if possible. } Thank you very much. } } } } Catherine Tang
Hi! This is Catherine Tang. I have got a lot of replies to my question yesterday. Actually I have no experience in LM processing. I know what to do now.
Fred Hayes wrote: =============================================== Does anyone know of a vendor that supplies lacey carbon coated Be grids? =============================================== SPI Supplies has been coating Be grids for some years now. There are several "secrets", one relating to the properties and characteristics of the Be grids you want to coat. Unlike grids of Cu, Ni, and Au which are electro deposited, Be grids can be made only by etching, and different etching protocols result in grids with differing degrees of surface protruberances (e.g. those little features that tend to tear a support film). You also have to be aware that the coating of Be grids is much more time consuming than the coating of Cu, Ni, or Au grids.
The "making" of lacey carbon and followed by carbon coating is not any different than for coating grids generally.
A final inspection of representative samples of the filmed grids is mandatory before shipping off to a customer because the "yield" is so much lower (when done on Be).
You can see additional information and prices on our website given below.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Some eagle-eyed environmentalists have discovered yet another Government conspiracy to conceal the true harmful nature of a chemical which commonly contaminates the environment with dire results.
Full details of this biohazard, dhihydrogen monoxide, can be found at the following website:-
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Colleagues,
Some eagle-eyed environmentalists have discovered yet another Government conspiracy to conceal the true harmful nature of a chemical which commonly contaminates the environment with dire results.
Full details of this biohazard, dhihydrogen monoxide, can be found at the following website:-
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Fred Hayes wrote: =============================================== Does anyone know of a vendor that supplies lacey carbon coated Be grids? =============================================== SPI Supplies has been coating Be grids for some years now. There are several "secrets", one relating to the properties and characteristics of the Be grids you want to coat. Unlike grids of Cu, Ni, and Au which are electro deposited, Be grids can be made only by etching, and different etching protocols result in grids with differing degrees of surface protruberances (e.g. those little features that tend to tear a support film). You also have to be aware that the coating of Be grids is much more time consuming than the coating of Cu, Ni, or Au grids.
The "making" of lacey carbon and followed by carbon coating is not any different than for coating grids generally.
A final inspection of representative samples of the filmed grids is mandatory before shipping off to a customer because the "yield" is so much lower (when done on Be).
You can see additional information and prices on our website given below.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
The other day I sent out directions for getting downtown from the airport via the local public transportation system, RTA. I meant to include for those who have internet browser capability the address for the RTA information system. Their site is at "http://little.nhlink.net/~rta/rtahome.html" (they also have a live picture of the Rock-N-Roll Hall of Fame). This site has all local public transportation schedules and many maps to help you get around the city using the public transportation system. The maps might also come in handy for those who are driving.
In the past year RTA has opened a new line for their rapid transit system which runs from Tower City out along the lake front to the Hall of Fame. This is their Water Front Line. I have not used this new line yet, so I don't know how to rate it.
Have a safe trip and I hope you enjoy your stay in Cleveland.
---------------------------------------------------------------- Dave Strecker mailto:dave.strecker-at-ab.com Rockwell Automation/Allen-Bradley Phone: (216)646-3250 Component Engineering ND246 Fax: (216)646-3416 1 Allen-Bradley Dr. Mayfield Hts., Ohio 44124 USA ----------------------------------------------------------------
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi! This is Catherine Tang. I have got a lot of replies to my question yesterday. Actually I have no experience in LM processing. I know what to do now.
We too are interested in calibration tolerences during our aperture QC procedures. Since the apertures we drill are as small as 5 microns and they are used in a number of applications as diverse as control jets for satelites and soldier production, tolernces can be very critical. We accept +/- 5% tolernces for magnifacation in our QC of our apertures, but I would be interested in any responses you get to this question.
We too are interested in calibration tolerences during our aperture QC procedures. Since the apertures we drill are as small as 5 microns and they are used in a number of applications as diverse as control jets for satelites and soldier production, tolernces can be very critical. We accept +/- 5% tolernces for magnifacation in our QC of our apertures, but I would be interested in any responses you get to this question.
Unsubscribe ===================================================== Marcelo Henrique Prado PEMM - COPPE/UFRJ Po.Box.:68505 Cidade Universit ria - Ilha do Fundao Rio de Janeiro-R.J. CEP.: 21941-900
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
On Wed, 6 Aug 1997, Tang Ee Koon wrote:
} I was given a block of paraffin wax with specimen embedded in it. I } would like to know how can I get away the wax to get out the specimen. } It is rather urgent and I would appreciate immediate reply if possible. } Thank you very much.
Would your specimen be harmed if you used solvent to remove the paraffin? Hexane, heptane or Petroleum Spirits 60-80, warmed slightly if necessary, would be suitable. They are not very toxic (but don't breathe too much), but they are highly flammable.
Several washings in a small quantity of solvent each time are MUCH MORE EFFICIENT than washing in one big lot of solvent (Bunsen's dilution law).
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello Catherine and all: Place the block in xylene and give it gentle agitation. Change the solvent a couple of times. Unless the block is very large you can de-wax the block overnight. Then dehydrate, opposite to hydration steps, but more rapidly. For the last step use single strength buffer and then fix the specimen in OsO4 and process as normal for EM specimens. Unfortunately specimens initially fixed for histology are never as good as those that were initially prepared for EM, in fact they are disappointing. However, they may show what is required and that is what matters most. Regards Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 400+ Links, MSDS ************************ http://www.proscitech.com.au
} } I was given a block of paraffin wax with specimen embedded in it. I } would like to know how can I get away the wax to get out the specimen. } It is rather urgent and I would appreciate immediate reply if possible. } Thank you very much. } } } } Catherine Tang
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
One of the major problems with crystals and crystal lattice specimens as a TEM test specimen is the need to determine the level of astigmatism in the image. As a novice service engineer needing a crystal lattice resolution picture you soon learn to place a little astigmatism in the image and hope! Its much easier than trying to find why you cannot resolve the lattice.
The best TEM test specimen to cover a wide range of operator levels is the holey carbon film. Tom Mulvey stated that "the minimum discernable fringe level is similar to the point to point resolution of the instrument"
A holey carbon film at 200,000X is a test for anyone, the object being to obtain the finest fringe that is even all round the hole. It tests the operators skills and their ability to truly observe an image. A through focal series is required but it is a waste of time doing this within two hours of switching on the kV as this will take time to stabilse. I have discussed heat gained in the tank needing to equal heat lost in order to obtain stability within the past few months on e-mail.
Correct the astigmatism at double the photo mag and then at the photo mag set the focus so that you just see an over or under focus fringe. Take a set of pictures through focus and then measure the centre of the black fringe to the centre of the white fringe ON THE NEGATIVE but only if the fringe is even. Fringes tell you a good deal about the instrument and its alignment (Reference Monitoring & Maintaining the TEM) Most people start off with about a 1 to 1.5nm resolution and if you are really good you may attain 0.45nm at which point the carbon structure starts to interfere with the fringe.
What do you learn.
1) You will see if the high voltage is stable - it is not perfect if you get focus drift, if you do not see some focus drift I would be surprised. 2) You will notice specimen drift when you first insert the rod 3) You will realise the importance of a good anticontamination device as the hole shrinks in size. 4) You will probably need more current - its that filament position again! 5) How difficult it can be at this level to correct astigmatism, you are looking for a fringe like a piece of cotton not a ships hauser. 6) Typical settings - 20-25uA emission, 0.5 to 1micron spot size, CII overfocus, eucentric point, 4 second exposure to test the machine (0.5 second exposures test nothing). 7) I do the test without an objective aperture so as to test the microscope, not the aperture cleanlyness.
Hope this helps please come back if you need more information.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Many people have problems obtaining information easily from their TEM and SEM, I see it almost every day. The area of instrument set up that causes more of these problems than any other is that of filament position.
Most people set the filament a long way from the cathode aperture and this leads to a long filament life but a low emission current. Moving the filament forward increases emission current but decreases filament life. With care and the additional use of the bias or emission control this forward movement of the filament will result in an improvement in performance - resolution. One problem, people place more credence in having a long filament life than in getting more from their microscope! Many buy a new machine because they feel the reason they cannot obtain the result they want is down to the microscope. Forget filament life if you want more from your machine, in my experience an instrument will be transformed if you push the gun a little harder. I could tell you so many stories where we have done this, much to the amazement of the owner and delight of the dissatisfied customer!
As for changing filaments the more you do it the less frightening it becomes, its really not difficult and to be honest if you want more from your microscope the cost is very little.
Why should people calibrate their microscope you may ask? If you do not take what is a pretty simple step how do you know that the instrument is working correctly? Is it not better to test the microscope via resolution, contamination and calibration than to have a hoard of customers banging on your door complaining that their results are poor; too late! Preventative maintenance, spotting problems at higher levels of operation than the normal in your laboratory, should be part of every well run laboratories routine. Spot the problems before they become a disaster and get them fixed before anyone else notices, thats they way to run a unit! It is no good saying the engineer does it one or twice a year, (does he really?) that gives a long period of uncertain operation and possible problems. In addition, if you learn to operate your microscope at higher levels than is the norm you improve your own techniques and powers of observation.
On one of my hobby horses, I believe that every laboratory should test its instruments and its operators routinely to see how they all perform. If you set a standard where you are testing in this way you instantly raise the levels of expertise and have standards which may be used over many months to prove that the laboratory is moving forward. Without such a regime laboratories stagnate! Everyone feels they do a great job but with no form of standard how do they know? We talk about Quality in electron microscopy in relation to how we set analytical standards and calibration; good! But an even more important question which is totally ignored is "how good are the operators?". I have said before microscopists are supposed to be scientists, real scientists would constantly test themselves!
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I was given a block of paraffin wax with specimen embedded in it. I would like to know how can I get away the wax to get out the specimen. It is rather urgent and I would appreciate immediate reply if possible. Thank you very much.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Since my days as a service engineer it seems to have been accepted that one could run a TEM within plus or minus 5% of the mag readout. You usually find that if the magnification is out across the range it is due to the high voltage level, specimen not at the eucentric point, or the final lens level. If the calibration is only out after a certain point it suggest that the lens which switches in at this point is at fault.
On the SEM unless you work with perfectly flat specimens magnification accuracy is a bit of a joke! Again it seems that if you check as many instruments as I see each year that 95% are within plus or minus 10%, and within plus or minus 5% X to Y. Either way its usually an engineer job to tune the scan circuits to improve the performance.
Peter Tarquinio Evex Analytical 857 State road Princeton, NJ 08540
----------
I've been ignoring the microscopy meeting that is coming to Cleveland and suddenly my boss said to me today, "hey, we should think about going to this thing next week". We work in Cleveland so thats not a problem. Could someone send me information like schedules and fees and seminars and classes, etc, etc, etc. Thanks. Mark Darus
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
JOB OPENING ANNOUNCEMENT!!!!!!!!!!!
Intel Corporation currently has an open position for a SIMS technician at its Santa Clara site's Materials Technology department. The SIMS group supports the Santa Clara site's development and fabrication facilities that produces state of the art microprocessors. The Materials Technology department's scope encompasses the whole process from silicon to a packaged device that includes process trouble-shooting, transfer and equipment qualifications, device failure analysis/fault isolation and new materials development to name a few. The department has a wide variety of state of the art instrumentation, including: SIMS, Auger, TEM, SEM/EDS, FIB, AFM, XRF, ICP-MS, FTIR, Raman, Pico and Nano Indentation.
Job Description:
The technician opening involves second shift operation of Quadrupole (Atomica) and Magnetic Sector (Cameca IMS- 6F) SIMS tools. SIMS is an Ultra High Vacuum analytical tool which uses ion beam sputtering combined with mass spectrometry detection to analyze dopant and contamination levels and distributions in patterned and unpatterned wafers. Duties will include data collection, data processing, report writing, interfacing with other engineers and customers, communicating results, and workflow duties to keep the lab running smoothly. The successful applicant must be self-motivated and capable of working with minimal supervision.
Qualification: An AA or BS degree with Engineering or physical science background or equivalent experience is required. Ability to work in a team oriented environment and good communication skills are critical. Experience with analytical equipment or Ultra High Vacuum or Vacuum systems is desired. Experience with SIMS and knowledge of Semiconductor processing methods is a definite plus.
Intel's industry leading total compensation package includes a competitive salary, stock options, annual employee bonus plan, bi-annual employee cash bonus payout, periodic paid sabbatical leave, and retirement plan. Intel is an equal opportunity employer.
Send resumes to or for more information contact:
Gabi Neubauer or Jerry Hunter Intel Corporation 2200 Mission College Blvd., M/S: SC2-24 Santa Clara, CA 95052
Phone: (408) 765-2241 or 765-2316 FAX: (408) 765-2393
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I've been ignoring the microscopy meeting that is coming to Cleveland and suddenly my boss said to me today, "hey, we should think about going to this thing next week". We work in Cleveland so thats not a problem. Could someone send me information like schedules and fees and seminars and classes, etc, etc, etc. Thanks. Mark Darus
We also use mag calibration tolerances of + or - 5%, and our company is ISO 9001 registered. However, I do not believe that ISO requires any specific tolerances for instrumentation; it simply requires that you determine what they are and conform to them. It is my belief that the "acceptable" tolerance is primarily dependent upon two things:
1) What can the instrument effectively yield? 2) What are your requirements? (i.e. How does the stated tolerance affect the quality of your data, and how critical is that?)
Just my thoughts on the topic.
Regards,
Bob ******************************* Bob Citron Chiron Vision Claremont, CA USA (909)399-1311 Bob_Citron-at-cc.chiron.com ******************************* ______________________________ Reply Separator _________________________________
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We have been using calibration tolerances of + or - 5% for both our magnification and spectrometer calibrations for quite some time now. Apparently these values have been established through supplier and user inputs quite some time ago. I am curious as to whether these values are consistent with those currently used or whether they are too loose. We are ISO 9002 registered and quality measures are very much a concern. TIA
Karpura Kommineni wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello everyone, } I am looking for a list of vendors of used light microscopes and parts. I } would like to have their phone number or other information that will help me } contact them. } You may reply directly to me. } Thank you } Sincerely } Karpura } kkommine-at-mbl.edu } Marine Biological Laboratory } Woods Hole Here are three vendors for used and new microscopes:
M2 Associates, Don Malatesta, 408-735-0495 McBain, 818-998-2702 Bender Associates, 602-820-0900
I hope these help.
Gary Liechty Allied High Tech Products, Inc. 2376 E. Pacifica Pl Rancho Dominguez, Ca. 90220 310-625-2466
I weep to hear that dihydrogen monoxide may be classified as a hazardous substance by the government authorities. Still, in some of our laboratories I have seen bottles of sand labled as hazardous material, presumably because they were labeled 'silica sand', and somehow or the othe anything with silicon or silica in it is considered taboo. Luckily, both are highly stable substances and can undoubtedly withstand the assult. However, I wonder when they'll start attacking hydrated hydronium hydroxide - a fearful sounding entity indeed, but one that is possibly more subject to having its reputation damaged than the others. If it were banned, we'd be in considerable trouble , indeed!
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
This subject is most distressing considering the material, also known as oxygen dihydride, has become so invasive in our daily routines. As many of you may be aware this materials the major ingredient in such pleasures as Guineas, Whatney's, John Courage, Old Speckled Hen,.... Need I say more?
Heath officials also recommend ingestion of at least 6 containers of DHMO/ODH daily - what are we to do?
Have a great day!! And thanks Mel for the insertion of a little humor!
Bob Craig
} ---------- } From: Melvyn Dickson[SMTP:M.Dickson-at-unsw.edu.au] } Sent: Thursday, August 07, 1997 1:01AM } To: microscopy-at-Sparc5.Microscopy.Com } Subject: New Toxic Hazard Shock } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Some eagle-eyed environmentalists have discovered yet another Government conspiracy to conceal the true harmful nature of a chemical which commonly contaminates the environment with dire results.
Full details of this biohazard, dhihydrogen monoxide, can be found at the following website:-
Steve Chapman's observations regarding SEM calibration are more than generous. Several years ago we did a calibration series on one of our SEM's using the Geller Standard and found that even though the instrument was well calibrated by the service engineer from the manufacturer - at the parameters that they specify for calibration - It was only valid for those conditions.
Any change in working distance, accelerating potential, beam current, etc., dramatically changed the validity of the calibrated values. For example, changing the working distance from 10mm to 30mm changed the magnification readout 20% from the actual standard values!!
These are things that are not usually mentioned or taught when dealing with instrument operation. A good point for any operator to be aware of when reporting "measured" values using a SEM.
As Steve also mentioned - forget about non-planar samples, or stage tilt.
Bob Craig OSRAM SYLVANIA Products inc. Lighting Research Center Beverly, MA 01915
} ---------- } From: Steve Chapman[SMTP:PROTRAIN-at-CompuServe.COM] } Sent: Wednesday, August 06, 1997 5:58PM } To: Wayne England; Msa link } Subject: Calibration tolerances } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I would like to know what software is available commercially AND will run on a PC for doing optical sectioning by digital deconvolution. Presumably, such software would be supplied with parameters such as x,y,z resolution, wavelength, N.A., etc.
David P. Bazett-Jones, Ph.D.
Professor Departments of Anatomy and Medical Biochemistry The University of Calgary 3330 Hospital Dr Calgary, AB T2N 4N1 Canada TEL: 403 220-3025, FAX: 403 270-0737 email:bazett-at-acs.ucalgary.ca
This is a multi-part message in MIME format. --------------04DFD1D8B6A8678A828255C7 Content-Type: text/plain; charset=us-ascii Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit Content-Transfer-Encoding: 7bit
Karpura Kommineni wrote: } } } Hello everyone, } I am looking for a list of vendors of used light microscopes and parts. I
-- -------------------------------------------------------------- Dr. Patrick L. Huddie (301) 725-2775 Fax (301) 725-2941 Microcosm, Inc., 9140 Guilford Road, Suite O, Columbia, MD 21046 e-mail phuddie-at-microcosm.com URL http://www.microcosm.com The Web is:"Vaster than empires and more slow" Andrew Marvell (1621-1678)
"Bother!" said Pooh as he was assimilated by the Borg "Bother!" said the Borg as they assimilated Pooh "Time for a little something" said the Borg "Oh dear!" squeaked Piglet while being assimilated by the Borg "Kanga is not a Borg identifier" squeaked the Borg
--------------04DFD1D8B6A8678A828255C7 Content-Type: text/x-vcard; charset=us-ascii; name="vcard.vcf" Content-Transfer-Encoding: 7bit Content-Description: Card for Patrick Huddie Content-Disposition: attachment; filename="vcard.vcf"
begin: vcard fn: Patrick Huddie n: Huddie;Patrick org: Microcosm, Inc. email;internet: phuddie-at-microcosm.com title: CEO x-mozilla-cpt: ;0 x-mozilla-html: TRUE end: vcard
The notice which follows was posted in March. Unfortunately the person to whom I offered the job has dropped out at the last minute. This job is available right now. If you are interested please contact me immediately. I will be at MSA in Cleveland next week - you can talk to me there or send me an e-mail message as soon as possible.
Microscopy and Computing
I am looking for a person to work on an exciting new project.
The appointment could be at the post-doctoral level or at other levels according to the background of the person appointed. In any case the post will be for three years.
The job is at the Materials Research Laboratory of the University of Illinois at Urbana.
The project is a joint enterprise involving Argonne National Lab, Oak Ridge National Lab, the Lawrence Berkeley Lab and NIST as well as the University of Illinois. The Project has the aim of developing a new kind of environment for electron microscopy and related techniques, in which the instruments can be operated remotely with the same effectiveness as they can be operated in the instrument room. More details of the project can be found at http://tpm.amc.anl.gov/MMC MMC is the abbreviation of the project name.
I am looking for someone who has familiarity with electron microscopy (preferably TEM) or a closely related technique - and who has well developed interests and experience in computing, particularly the interfacing of instruments for computer control and/or the networking of images.
Will any one interested please contact me right away. We would like the job to be started as soon as possible. ** Alwyn Eades Center for Microanalysis of Materials University of Illinois at Urbana-Champaign Phone 217 333 8396 Fax 217 244 2278 eades-at-uimrl7.mrl.uiuc.edu (NB those are letter l not ones) **
EMAG '97 Conference and trade exhibition ----------------------------------------
The biennial EMAG conference is being held in the Cavendish Laboratory, Cambridge from Tuesday 2 to Friday 5 September 1997. The principal aim of the conference is to promote and discuss recent advances in electron microscopy and related analytical techniques. Accompanying the conference is a major international trade exhibition and at present there are still a small number of stands available to potential exhibitors.
Conference sessions will be held on: microanalysis, semiconductors and superconductors, high resolution electron microscopy, ceramics/interfaces, electron crystallography, EELS, materials analysis, new instrumentation, advanced scanning probe techniques, microscopy of catalysis, intermetallics and advanced SEM and surface science.
For information on the conference see http://www.iop.org/IOP/Confs/EMAG/
For information on the exhibition or to book a stand at the exhibition see http://www-hrem.msm.cam.ac.uk/emag97/ or contact Chris Boothroyd, cbb4-at-cam.ac.uk Tel: +44 1223 334564 Fax: +44 1223 334567
At 08:36 AM 8/7/97 -0400, you wrote: -=-snip-=- } Any change in working distance, accelerating potential, beam current, } etc., dramatically changed the validity of the calibrated values. For } example, changing the working distance from 10mm to 30mm changed the } magnification readout 20% from the actual standard values!! } } These are things that are not usually mentioned or taught when dealing } with instrument operation. A good point for any operator to be aware of } when reporting "measured" values using a SEM. } } As Steve also mentioned - forget about non-planar samples, or stage } tilt. -=-snip-=-
Hello List,
Many modern SEMs take into account HT and working distance and adjust the mag accordingly (I know ours have for at least 10 years). Any given magnification on these scopes is usually +/- 5% of the displayed mag although there may be more than 5% difference compared to another mag.
Dave Harrison Site Manager JEOL USA, INC
"The trouble with America isn't that the poetry of life has turned to prose, but that it has turned to advertising copy." Louis Kronenberger
Does anyone have resin embedded, encephalitis infected tissue that they would be willing to share? After our son's illness, my husband and I started a grassroots organization (which appears in our email). Consequently, I decided to postpone med-school for two years to do some serious research. My dilemma is that I am attending a State funded school that will not permit me to generate my own tissue samples. The reasons: Animal Rights Committee regulations, the nature of the pathogen, and, of course, funding. The University is willing to back the endeavor--once the resin blocks are in hand. Any information would be greatly appreciated.
Sincerely, Debra Caires San Jose State University Biology Department One Washington Square San Jose, CA 95192 (408) 298-2060
As Dave Harrison states, all modern SEMs compensate for variables such as Working Distance, KV, Spot Size, etc.
Older SEMS had Mag readouts with only most significant digits and large steps between values.
Newer SEMS have more accurate mag readouts with smaller steps such as 1000, 1001, 1002, etc.
Many modern instruments have a Magnification Calibration procedure which adjusts the Mag to the Calibration sample used. If this method is used, and then the Mag is measured with the same sample, errors of less than 1% are possible. However, if you calibrate with one sample and then measure with another, then the Mag is only as good as the accuracy of the samples.
One of the funniest things that I have seen is safety data for water in one of these books of chemical hazards. It had phrases like: if splashed in eyes rinse with plenty of WATER, and in case of a spillage wash down drain with plenty of WATER!!
It makes you wonder sometimes.
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
On Fri, 8 Aug 1997, Ian MacLaren wrote: } } One of the funniest things that I have seen is safety data for water in one } of these books of chemical hazards. It had phrases like: if splashed in } eyes rinse with plenty of WATER, and in case of a spillage wash down drain } with plenty of WATER!!
Ian. here is the copy of that text, sent on this list a few month ago:
Hello all
This is a slightly facetious input re. safety data sheets!
In the UK there is a very useful pair of publications from BDH, a chemicla supply company (used to be known as British Drug House, I believe). These are collcted data sheets. Yes folks, there is one for water. Here are a few salient points which all users of the substance should bear in mind at all times:
1. colourless liquid - maybe that implies it is rare and difficult to see if dropped! 2. Against solubility in water: miscible in all proportions. 3. fire and explosion hazard: not applicable (thats a relief - although if it caught fire I suppose one could foolishly attempt to extinguish with more water?). 4. Health hazard: no significant hazard expected, may be irritating to the eyes. 5. Toxicity: no data. 6. Carcinogenicity: no evidence of carcinogenic properties. 7. First aid - eyes: irrigate thoroughly with water(!) 8. First aid - lungs: remove from exposure. 9. First aid - skin: wash off thoroughly with soap and water (work that one out!). 10. First aid - mouth: wash out thoroughly with water. In severe cases obtain medical attention(!) 11.Reactive hazards: violent reaction with acyl halides, alkali and alkaline earth metals .... 12. Spillage disposal: wear appropriate protective clothing - listed are gloves, goggles, face shield and protective apron. Luckily, respirators are not needed.
I have an MSDS for soap. Skin contact requires washing with soap and water. However, it is not clear if you can use the SAME soap or if you need to use a DIFFERENT soap. Since this problem is unresolved, I just hope and pray I never get any soap on me. What would I do?
Many :-)
} ------------------------------------------------ } Opinions or statements expressed herein, rational or otherwise, do not } necessarily reflect those of my employer. } } Harold J. Crossman } OSRAM SYLVANIA INC. } Lighting Research Center } 71 Cherry Hill Dr. } Beverly, MA 01915 } Phone: (508) 750-1717 } E-mail: crossman-at-osi.sylvania.com } } Our web sites: www.sylvania.com } www.siemens.com } -- } } "Crossman, Harold" {crossman-at-osi.SYLVANIA.com}
Let me describe the procedure we used to determine variations in magnification values, etc.
The instrument was calibrated against the Geller Standard by the manufacturers service engineer using their procedures. Magnification, x vs y, image squareness, etc., - all within =B1 1%. In all cases the = beam was orthogonal to the standard surface. Changing the working distance from 10mm to 30mm, refocussing on the standard, removing hysteresis from the column (i.e., "ringing" the lenses), adjusting the magnification to the same value as at calibration, recording an image, measuring the same pair of lines on the standard and comparing these values with the original calibration image resulted in an error of -20%. We also did this procedure at several intermediate working distances and found the variation was not linear with working distance as well.
Going to shorter working distances had a positive effect, but no where near as large as with longer working distances, i.e., the rate of change in off-calibration per mm change in working distance was less for shorter working distances.
Nothing was changed except the working distance and the difference in lens and scan coil current required to produce crossover on the standard at the lower position. The beam current was not adjusted to compensate for the difference in lens current (which normally would not be done unless doing "quant" EDS).
I know nothing about, nor do I really care about, what compensating circuits are supposed to do, etc., just that when parameters are changed one must be aware that readouts may not be telling the "truth and nothing but the truth."
It is my opinion that many SEM operators at not aware of these little vagaries and should be made aware of them and realize that those neat little digital readouts might not be accurate under conditions different from the instruments calibration parameters.
Bob Craig OSRAM SYLVANIA Products Inc. Lighting Research Center Beverly, MA 01915
Hi, As organiser of the forthcoming First Electronic Analytical Chemistry Conference, I would like to invite someone well know in the field to present an interesting talk on Microscopy. Since this is not my field I would like suggestions as to who to approach with an invitation.
The conference will take place over the internet in early November and will take the form of previous electronic conferences which were very successful.
ECCC1 & ECCC2 First and Second Electronic Computational Chemistry http://hackberry.chem.niu.edu:70/0/ECCC/homepage.html
ECHET96 Electronic Conference on Heterocyclic Chemistry http://www.ch.ic.ac.uk/ectoc/ehet96/
During the conference interaction, presentations and discussions will take place via the Internet using a Java-based virtual conference centre, WWW-based discussion forums and an electronic mailing list.
The Iowa Microscopy Society will hold its annual symposium on Thursday, September 25, 1997 at the Eckstein Medical Research Building on the University of Iowa campus in Iowa City.
TENTATIVE MEETING AGENDA September 25th
7:30-9:00am Registration Poster and display set-up
8:00 Welcoming Remarks
8:20 "Morphological Characterization of the Metastatic Mary Hendrix Department of Anatomy, University of Iowa
8:50 "HIV-Induced T-cell Syncytia, In Vivo and In Vitro, are Motile, Invasive and Destructive" Karla Daniels Department of Biological Sciences, University of Iowa
9:35 "To be announced" Charles Gross Department of Microbiology, University of Iowa
10:05 BREAK
10:30 "Fractility: Correlation of Images with Transport Characteristis of Exchange Polymer Composites" Johna Leddy Department of Chemistry, University of Iowa
11:00 "Applications of Multiphoton Excitation Imaging." Victoria Centonze-Frohlich IMR, University of Wisconsin
12:00 LUNCH
1:00 "Same Cell Correlative Video Enhanced Light and 3-D Electron Microscopic Studies of Mitosis in Vertebrate Cells" Conley L. Rieder NIH
2:00 "The Effect of Mutation of COMP on Calcium Deposition in Chondrocytes" Jeff Stevens Department of Orthopedic Surgery, University of Iowa
2:30 BREAK
3:00 "BCL-2 Expression Alters Targeting of Viral Products in Insect Cells" David Murhammer Department of Chemical Engineering, University of Iowa
3:30 "Advances in Ultra-Small Gold Probes in Light and Electron Immunocytochemistry and In-Situ Hybridization" Peter Van de Plas AURION, The Netherlands
4:30 "Fluorescent Annual Banding in Speleothems, Applied to Paleoclimatology" Christopher Shorey Departmnt of Geology, University of Iowa
5:00 IMS Business Meeting
5:15 RECEPTION
Symposium costs are:
Preregistration: Meeting day registration: regular-$8.00 regular-$12:00 student-$5.00 student-$7.00
For a registration form or further details please contact Kathy Walters (addresses below).
IMMUNOCYTOCHEMISTRY WORKSHOP
In addition to the symposium, a workshop is offered on the following Friday and Saturday (September 26th and 27th). Peter van de Plas will provide a general lecture of immunocytochemical techniques and hands on instruction. For more details please visit the CMRF website at:
http://www.uiowa.edu/~cemrf
POSTER COMPETITION
Three cash prizes in two catagories will be awarded to outstanding participents. Applications for submitting posters to the meeting are now being mailed. If you would like an application, or would like to be added to our mailing list, please contact Kathy Walters at the address below.
Kathy Walters / / Research Assistant III / /\ Center for Microscopy Research / /\ \ University of Iowa /_/ \ \ 85 EMRB ____ ((O)) Iowa City, Iowa 52242 | | / / || / / email: kwalters-at-emiris.iaf.uiowa.edu ----------- fax: (319)335-8049 ------------- www: http://www.uiowa.edu/~cemrf
I would be grateful if someone could point me to a reference on the imunno electron microscopic localization of carbonic anyhdrase in mamallian tissue. No hurry since I am leaving for Cleveland on Sunday. See you all there.
Greg Erdos ******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab PO Box 118525 Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
Can anyone help me find a supplier for EMHOPC (ethyl-1-methyl-4-hydroxy-oxo-3-pyrroline-3-carboxylate)? It is a ferric ion chelating agent that has been reported to give permanent staining of ATPase activity in skeletal muscle fibers, ref: Hawcroft DM ,Ball MT A new chelating method for determining ATPase activity in skeletal muscle. Biotechnic & Histochem 1996; 71 (2): 88-91. I have tried contacting the authors by mail, but no reply. Perhaps some of my fellow Brits can help, as Drs Hawcroft and Ball are employed in the Department of Pharmaceutical Sciences, De Monfort University, Leicester, England. Thanks, in advance, for any assistance. Ronnie Houston Director Cytochemistry & Molecular Pathology Texas Scottish Rite Hospital for Children Dallas, TX 75219 USA
Applied Precision 8505 SE 68th Street Mercer Island, WA 98040
Phone: 206-236-0704 Fax: 206-232-4184
-------------------------------------------------------------- Dr. Patrick L. Huddie (301) 725-2775 Fax (301) 725-2941 Microcosm, Inc., 9140 Guilford Road, Suite O, Columbia, MD 21046 e-mail phuddie-at-microcosm.com URL http://www.microcosm.com The Web is:"Vaster than empires and more slow" Andrew Marvell (1621-1678)
-------- REPLY, Original message follows --------
I run the Electron Microprobe Lab at Arizona State University, where we have A JEOL 8600 probe with (Noran Tracor) 5500 Series II EDS system and 5600 PAC automation system. The Vista image analysis software works fairly well - however, Noran did not engineer the capability to export the images to the outside world in a real-world format. We have found a way around this problem by using the } XI 2 Terminal Emulator supplied by Noran. The image has to be transmitted as a TEXT file, and the receiving PC using SmartTerm receives it as a text file. It takes five minutes to transfer a 512x512 gray-scale image, such as SEI and BEI. The PC disk then has to be transferred to a MAC to convert it to a standard TIFF file through the NIM Image program. While it does work, it is understandably slow and a real pain.
The problem that I am trying to resolve now is figuring out how to transfer EDS x-ray dot maps. The closest I've coming to succedding is to collect the map for a single element, convert it to a binary image, use } XI 2 to transmit as a binary image to The PC using SmartTerm to receive it as a binary image. I get an image on the MAC, but it is barely visible. The dots are very faint gray instead of white, and the background is an intermediate to light gray instead of black. I had stored the binary image in Binary 1, and left the other Binary bins blank. I had also tried copying the faint image in Binary 1 into Binary 6 thru 7, so that all bins contained the same image. No Luck. Finally, I tried transmitting the lone Binary image in Binary 1 and transmitting it as a binary image to SmartTerm as a TEXT file. Again, no luck.
I seem to have run out of ideas. I have called Noran, but they had no suggestions. Todd Jarlsberg had been working on this problem and seemed to have a pretty good handle on it, but left Noran a couple of years ago.
Has anyone out there been down this same road, and if so have you come up with a solution, or at least have any suggestions. We would prefer if at all possible to resolve this problem without having to add upgrades to the system, money being tight as it is these days, but that may be unavoidable.
Thank you for your ideas and efforts.
Jim Clark e-mail: jclark-at-asu.edu Probe Lab: 602/965-61720
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } JOB OPENING ANNOUNCEMENT!!!!!!!!!!! } } Intel Corporation currently has an open position for a SIMS technician at its } Santa Clara site's Materials Technology department. The SIMS group supports } the Santa Clara site's development and fabrication facilities that produces } state of the art microprocessors. The Materials Technology department's scope } encompasses the whole process from silicon to a packaged device that includes } process trouble-shooting, transfer and equipment qualifications, device failure } analysis/fault isolation and new materials development to name a few. The } department has a wide variety of state of the art instrumentation, including: } SIMS, Auger, TEM, SEM/EDS, FIB, AFM, XRF, ICP-MS, FTIR, Raman, Pico and Nano } Indentation. } } Job Description: } } The technician opening involves second shift operation of Quadrupole (Atomica) } and Magnetic Sector (Cameca IMS- 6F) SIMS tools. SIMS is an Ultra High Vacuum } analytical tool which uses ion beam sputtering combined with mass spectrometry } detection to analyze dopant and contamination levels and distributions in } patterned and unpatterned wafers. Duties will include data collection, data } processing, report writing, interfacing with other engineers and customers, } communicating results, and workflow duties to keep the lab running smoothly. } The successful applicant must be self-motivated and capable of working with } minimal supervision. } } Qualification: An AA or BS degree with Engineering or physical science } background or equivalent experience is required. Ability to work in a team } oriented environment and good communication skills are critical. Experience } with analytical equipment or Ultra High Vacuum or Vacuum systems is desired. } Experience with SIMS and knowledge of Semiconductor processing methods is a } definite plus. } } } Intel's industry leading total compensation package includes a competitive } salary, stock options, annual employee bonus plan, bi-annual employee cash } bonus payout, periodic paid sabbatical leave, and retirement plan. Intel is } an equal opportunity employer. } } Send resumes to or for more information contact: } } Gabi Neubauer or Jerry Hunter } Intel Corporation } 2200 Mission College Blvd., M/S: SC2-24 } Santa Clara, CA 95052 } } Phone: (408) 765-2241 or 765-2316 } FAX: (408) 765-2393 }
A colleague is seeking a means for tranferring images recorded on 2" video floppies to more standard digital format. These video images were generated/stored with a Hitachi VX-100A Video Floppy System. However, we don't have the system here; just the disks. In fact, we can't even find information on this system on Hitachi's website.
So my questions are: Has anyone ever heard of this Hitachi video floppy system? What is required to read the disks? Most importantly, does anyone know of a local (Minnesota) system that could read these disks and transfer them to a computer? Even transfer to VHS format could be useful, if nothing else is available.
Thanks as always for your help and suggestions! Karen
-- Karen Zaruba kszaruba-at-mmm.com 3M Company, 3M Center Bldg. 270-1S-01 St. Paul, MN 55144 "The opinions stated above are my own, not necessarily 3M's"
I wish to hear from fellow users of the JEOL 6400F, if there are any out there. Specifically, I am curious about your experiences running the system at low kV, 600V to 3kV accelerating voltage. Have you experienced any image instabilities? Running in "super rapid", I have seen the entire image shift about 1/4 to 1/2 inch and then return to the original position. In photos, this shows up as a band in the image that is shifted sideways. Another anomaly I have seen is the brightness jumping up and down. In photos this shows up as brighter and darker bands horizontally across the photo. If you have seen these effects, has there been any solution or explanation? (Other than it is being caused by fields, probably from Alfa Centari! :-) )
Used Microscopes can be obtained via several sources on our Services Page. You could also place a request for individual mcroscopists who might be selling off their instruments on our Forum pages... all at: -
http://www.microscopy-uk.org.uk
These pages, and the site, predominantly cater for amateur (optical) microscopists... but of course - everyone is welcome!
I am not familiar with the Tracor/Noran format, so may may wish to contact John Mansfield at U-Mich. He has a Tracor and has done networking with it to other platforms. I suppose he knows of a method or utility for handling maps. There are converter utilities for many things thru the file archives. But again, John will have to point you to the right files. You can start with the URL ftp://www.amc.anl.gov/AMC-3/ANLSoftwareLibrary/ and go from there.
I would hope you can pick off the map parts and transfer them over as an image without going binary first. You would probably need to normalize the grayscale after transfer. Our Link stores the count as the grayscale so that we only hit 255 on very long maps. Most of our maps run from 0 to less than 40. Many packages can remap that to a 0 to 255 range so that the trends are visible.
Good hunting.
At 09:16 AM 8/8/97 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am trying to assist a research group interested in looking at nanoparticles. Some are made of silver iodide and others are made of cadmium sulfide, sometimes with a little zinc. They are supposed to be about 5 micrometers in size. They do the synthesis and some sorts of analysis, infrared spectra, I think. They come to my TEM to check on the size.
Well, I am having a hard time seeing them. I tried a drop of suspended particles on a formvar coated grid, a technique that usually works OK. I could hardly see any particles in the TEM. I saw a few particles that might have been what they are looking for, but they were few and far between. I suggested that the concentration might be low, they said, no, should be lots in there.
Without going into lots of details here, I promised them I would try to find out how to look at their particles. I said I would try a question to the list. Soooo, if you think of any helpful hints send them my way.
Thanks.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
You know those 5 micrometer particles I couldn't see? Well, they are really supposed to be 5 nannometers (duh). You see I was so excited about getting ready to go to Cleveland that I lost my head. Hopefully nobody will notice this dumb mistake until after I get back from the meeting!
See (saw?) you there.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
Dear Jon, With particles that huge, the suspension should be cloudy if it is of any concentration. In fact, you should be able to look at a dried suspension on polished graphite in the SEM and check the composition by EDX. Just because their recipe says it should produce these particles, doesn't mean it necessarily will. I have been looking for spheres of Ni, 50 nanometers diameter, that the Chemical Engineering Department has been trying to make through two Master's theses. So far we have seen maybe five. Can you use a carbon film? It is clearer. With those heavy elements, imaging a dried suspension on a formvar should show the particles clearly at 100kV on the TEM. Do you have EDX on your TEM? I have looked at silver chloride crystals, silver spheres 30 nm. in diameter and the aforementioned nickel spheres. The main problem is the other dissolved solids in the suspension obscuring the whole grid, in which case the whole grid is dark and blurry. You wrote:
} Help! } } I am trying to assist a research group interested in looking at } nanoparticles. Some are made of silver iodide and others are made of } cadmium sulfide, sometimes with a little zinc. They are supposed to be } about 5 micrometers in size. They do the synthesis and some sorts of } analysis, infrared spectra, I think. They come to my TEM to check on the } size. } } Well, I am having a hard time seeing them. I tried a drop of suspended } particles on a formvar coated grid, a technique that usually works OK. I } could hardly see any particles in the TEM. I saw a few particles that might } have been what they are looking for, but they were few and far between. I } suggested that the concentration might be low, they said, no, should be } lots in there. } } Without going into lots of details here, I promised them I would try to } find out how to look at their particles. I said I would try a question to } the list. Soooo, if you think of any helpful hints send them my way. } } Thanks. } } Jonathan Krupp } Microscopy and Imaging Lab } University of California } Santa Cruz, CA 95064 } (408) 459-2477 } FAX (408) 429-0146 } jmkrupp-at-cats.ucsc.edu
Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
} We are shopping for a TV rate camera to interface to our Hitachi H-600 TEM } via a 35 mm camera port (scope is equipped with STEM). We plan to use } this for teaching and when more than one person (i.e. operator and } researchers) are working at the scope, not for acquisition of high-quality } digital images. To the best of my knowledge these systems work as } follows: a small phosphor screen will be moved into the beam path, a } camera is focused on this screen and the image is displayed on a } small B&W monitor. I have found two vendors so far (Fullam and Gatan), } but our purchasing department wants more. If you know of any other } vendors for this kind of equipment - or are vendors of it - please reply } directly to me by e-mail. } } Thanks in advance for any replies. } } Heather Owen
If you receive, or can get hold of, a copy of Microscopy & Analysis, there are adverts from the main companies plus a buyers guide - if you use the buyers guide, you'll get all the info for a specific product area sent to you.
I have an interest here, as Technical Editor.
You also need to distinguish between lens coupled systems, which is what you describe, and directly coupled system, where the camera itself is placed inside the vacuum of the TEM. A transmission phosphor (YAG) is used, which is coupled by a fibre optic plate to the camera (usually a CCD).
Generally, you will find that directly couple systems are more efficient a collecting the light from the phosphor than lens coupled systems.
Regards,
-- Dr L. P. Stoter Technical Editor, MICROSCOPY & ANALYSIS Technesis M&A web site - 17, Rocks Park Road http://www.microrgc.demon.co.uk Uckfield, E. Sussex email: LPS-at-teknesis.demon.co.uk TN22 2AT Phone: +44 (0)1825 766911 United Kingdom Fax: +44 (0)1825 766911
Hello - I don't know about silver iodide, but I've been assisting a bit in= using TEM to look at 2-5 nm diameter CdS particles stablized using either= low molecular weight ligands, or block copolymers. They show a lot less BF= contrast than gold, say. If there are a lot of them, close to Gaussian= focus the overall texture looks more like amorphous carbon at high defocus= than a neat array of distinct little round balls (and it looks like a= complete mess at high defocus, especially given all the organic stuff in= there).=20 So, assuming you aren't using EDX to check for Cd, and that you are= not capable of imaging CdS lattice fringes, then it might be that you're= just not seeing the trees for the wood. In that case, try diluting the= suspension and picking the particles up on a thin carbon film (rather than= formvar). It's also an idea to use a holey carbon film - then you should be= able to see some of the particles hanging over the edges of the holes, even= if they are not showing up well against the carbon.=20 In my experience, if the light absorbtion data say you have a lot of= little particles in suspension, then with the droplet method you generally= end up with a lot of little particles on the carbon film (especially with= CdS, which seems to be everybody's favorite system). In such cases we go= straight to lattice imaging on an EM 430/LaB6 to get the particle sizes (so= that it doesn't particularly matter if the low resolution BF images don't= show a great deal). If the nanoparticles all decide to clump together and= float away during the drying step, then you can usually see that happening= in the optical microscope, and freeze drying is in order. If they're= already clumped together then the suspension will be cloudy and the= chemists need to get their act together. =20
Pr=E9nom Nom: Christopher John George Plummer Institut: Laboratoire de Polym=E8res, D=E9partement des Mat=E9riaux EPFL: Ecole Polytechnique F=E9d=E9rale de Lausanne CH-1015 Lausanne t=E9l: (+41 21) 693 28 56 email: christopher.plummer-at-lp.dmx.epfl.ch
Characterization of 5nm-sized AgI and Cd(Zn)S particles requires TEM and STEM techniques with an appropriate resolution and a higher electron beam intensity, which can be readily obtained with microscopes equipped with LaB6 or field emission guns. It may need also preparing thinner supports than conventional formvar films. Better results one can achieve by the use of amorphous carbon films and carbon holey films up to 2nm in thickness. Here are some references for the reproducible protocol for making such films:
Procedures in Electron Microscopy /Eds. A.W.Robards and A.J.Wilson. J.Wiley&Sons, Chichester, 1993,pp.4:6.15-6.22. W.Baumeister and J.Seredynsky/Micron, 1976, v.7, pp.49-54.
Some problems may cause aggregation and coalescence of particles during deposition on film-supports. Therefore particles in a suspension of an appropriate concentration should be stabilized by adding of a protective polymer such as polyphosphate or by suitable complexing ligands.
In my experience, electron beam-induced decomposition and related phenomena especially in the case of AgI nanoparticles resulting in quick reduction to silver with elimination of halide, melting, evaporation and recrystallization of particles may significantly hamper studies. For such observations it would be necessary to use a cryo-cooling at liquid nitrogen temperature. One can also reduce damage, modification and drift effects by working fast and/or using minimal electron doses.
Regards,
Vladimir Oleshko ********************************************************** V.P. Oleshko, Ph. D. e-mail:oleshko-at-uia.ua.ac.be Micro-and Trace Analysis Centre Tel.:+32-3-820.23.64 Chemistry Department FAX :+32-3-820.23.76 University of Antwerp (UIA) Universiteitsplein 1 Antwerpen-Wilrijk B-2610 Belgium ***********************************************************
On Fri, 8 Aug 1997, Jon Krupp wrote: } Help! } } I am trying to assist a research group interested in looking at } nanoparticles. Some are made of silver iodide and others are made of } cadmium sulfide, sometimes with a little zinc. They are supposed to be } about 5 micrometers in size. They do the synthesis and some sorts of } analysis, infrared spectra, I think. They come to my TEM to check on the } size. } } Well, I am having a hard time seeing them. I tried a drop of suspended } particles on a formvar coated grid, a technique that usually works OK. I } could hardly see any particles in the TEM. I saw a few particles that might } have been what they are looking for, but they were few and far between. I } suggested that the concentration might be low, they said, no, should be } lots in there. } } Without going into lots of details here, I promised them I would try to } find out how to look at their particles. I said I would try a question to } the list. Soooo, if you think of any helpful hints send them my way. } } Thanks. } } Jonathan Krupp } Microscopy and Imaging Lab } University of California } Santa Cruz, CA 95064 } (408) 459-2477 } FAX (408) 429-0146 } jmkrupp-at-cats.ucsc.edu } } }
{bold} {color} {param} FFFF,0000,0000 {/param} {bigger} {bigger} PHILIPS EM201 FOR SALE
{/bigger} {/bigger} {/color} {/bold} A Philips EM 201 transmission electron microscope in excellent condition is available for immediate sale. The instrument had been used basically by one person, and is covered by service contract. A new plate camera was installed last year. Asking price is $3500. Crating and shipping charges would be buyer's responsibility. If interested, please contact:
Dr. Cornelia Farnum
Anatomy Department, College of Veterinary Medicine
The University of Glasgow and The Queen's University of Belfast are seeking 2 Postdoctoral Research Assistants for the following=20 research project.
THE USE OF XANES AND ELNES FOR THE CHARACTERISATION OF STABILISED ZIRCONIA= =20
The University of Glasgow and the Queen's University, Belfast have been awarded a major EPSRC grant to investigate and develop the use of near edge fine structure in X-ray absorption spectroscopy (XANES) and electron energy-loss spectroscopy (ELNES) for the characterisation of commercially important zirconia-based materials. This multidisciplinary project involves researchers in four universities, as well as researchers at the Daresbury Laboratory and in two UK companies, MEL Chemicals Ltd and Johnson Matthey= Ltd.
We are seeking two outstanding postdoctoral research assistants (PDRA) to work on this project. PDRA1 will be based in Glasgow for 30 months and will carry out the experimental work using the analytical electron microscopy facilities at Glasgow and the synchrotron facilities at Daresbury. PDRA2 will be based in Belfast for 12 months and in Glasgow for the following 18 months and will develop and apply theoretical models for calculation of ELNES & XANES in defective zirconia materials. Both posts require candidates with a practical approach to problem solving and should appeal to those with a background in solid state chemistry/condensed matter physics/materials science. For PDRA1, experience in electron microscopy, especially electron energy loss spectroscopy, would be an advantage. For PDRA2, experience with first principles band structure calculations is essential and a background in the theoretical interpretation of spectroscopic techniques such as ELNES and XANES would be highly desirable, as would a knowledge of many-body physics.
Initial salary will be on the RA1A scale up to =A316,927 per annum,= depending on qualifications and experience. Applicants should send two copies of their CV along with the names and addresses of two referees to Dr David McComb, Department of Chemistry, University of Glasgow, Glasgow G12 8QQ, UK.
Informal enquiries and requests for further details can be made to Dr Alan Craven (0141 330 5892, a.craven-at-physics.gla.ac.uk), Dr David McComb (0141 330 4486, davidm-at-chem.gla.ac.uk) or Prof. Mike Finnis (01232 335330, M.Finnis-at-qub.ac.uk). Further details can also be obtained on the following websites; www.chem.gla.ac.uk, www.ssp.gla.ac.uk, titus.phy.qub.ac.uk.
The closing date for applications is 12 September, 1997. ----------------------------------------------------------------------------= ---- Dr David W McComb Department of Chemistry University of Glasgow Glasgow G12 8QQ U.K.
I once worked on solar-cell multilayers, and found that almost all of those compounds, including CdS, were very beam sensitive. They evaporate and leave nothing during observation, or re-deposite on the specimen and destroy any nice trasparent areas in ~10 sec. at 120kev. I believe you may be using a higher kev (200kev?) to see 5nm particles. It seems to me that the particles could be gone before you saw them.
If you just need to know the particle size, a very thin carbon coating on your specimen may help you out. Otherwise, you may send me the particles. This company accepts TEM/OM consulting services of particles, thin films and bulks of any materials.
Chao-Ying Ni, PhD Manager of Microscopy Dept. Batta Laboratories Inc. 6 Garfield Way Delaware Industrial Park Newark, DE 19713
On Fri, 8 Aug 1997, Jon Krupp wrote:
} Date: Fri, 8 Aug 1997 16:33:31 -0700 } From: Jon Krupp {jmkrupp-at-cats.ucsc.edu} } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Imaging AgI & CdS nanoparticles } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Help! } } I am trying to assist a research group interested in looking at } nanoparticles. Some are made of silver iodide and others are made of } cadmium sulfide, sometimes with a little zinc. They are supposed to be } about 5 micrometers in size. They do the synthesis and some sorts of } analysis, infrared spectra, I think. They come to my TEM to check on the } size. } } Well, I am having a hard time seeing them. I tried a drop of suspended } particles on a formvar coated grid, a technique that usually works OK. I } could hardly see any particles in the TEM. I saw a few particles that might } have been what they are looking for, but they were few and far between. I } suggested that the concentration might be low, they said, no, should be } lots in there. } } Without going into lots of details here, I promised them I would try to } find out how to look at their particles. I said I would try a question to } the list. Soooo, if you think of any helpful hints send them my way. } } Thanks. } } Jonathan Krupp } Microscopy and Imaging Lab } University of California } Santa Cruz, CA 95064 } (408) 459-2477 } FAX (408) 429-0146 } jmkrupp-at-cats.ucsc.edu } } }
Darrell Miles asked about shifting images on a JEOL 6400F:
We had similar problems on our JEOL 6000F (sudden lateral movement of the image). This was eventually, after MUCH investigation by all concerned, traced to very low frequency ( } } 1hz) magnetic fields. In our case the probable source is the railway approx 250m away from the microscope (but we have not ruled out Alpha Centaurus!). We have measured magnetic intensities up to 4mG, running in X, Y and Z directions. Due to the low frequency (approximating DC fields) it is not possible to monitor these fields with the normal multiturn coil connected to an oscilloscope, which is why it took us so long to identify the problem. Field emission microscopes seem to be much more prone than tungsten filament microscopes to magnetic interference (the other two scanners in the immediate surroundings are tungsten filament machines, JEOL 840 and 5800LV) and are much less affected. Shielding is not effective for the attenuation of these low frequency fields. Our solution was the installation of field cancelling equipment - expensive, somewhat of a nuisance, but effective. Darrell's second problem could be due to sparking on the SE detector scintillator - one way of checking this is to electrically connect the aluminium coating of the scintillator disk to the scintillator holder (a VERY small dab of carbon paste usually does the trick). If the brightness variation is seen also with the BE detector, the source of the variation is of course elsewhere - possibly in the noise canceller??.
} I wish to hear from fellow users of the JEOL 6400F, if there are any out there. } Specifically, I am curious about your experiences running the system at low } kV, 600V to 3kV accelerating voltage. Have you experienced any image } instabilities? Running in "super rapid", I have seen the entire image shift } about 1/4 to 1/2 inch and then return to the original position. In photos, this } shows up as a band in the image that is shifted sideways. Another anomaly } I have seen is the brightness jumping up and down. In photos this shows } up as brighter and darker bands horizontally across the photo. If you have } seen these effects, has there been any solution or explanation? (Other than } it is being caused by fields, probably from Alfa Centari! :-) ) } } Thanks, } Darrell
Prof Jan Coetzee Head: Unit for Electron Microscopy Tel:+27-12-420-2075 University of Pretoria Fax:+27-12-342-1738 Pretoria 0002 Internet:janc-at-ccnet.up.ac.za South Africa http://www.up.ac.za/science/electron/emunit1.htm
I have a Matrox Meteor imaging board and would like to acquire RGB images via a JVC TK1070E CCD camera fitted to a Zeiss confocal microscope. Does anyone know of any shareware (or fairly cheap) image acquisition software that I could use to view and acquire images, preferably as TIFF image ?
Regards
Mark Auty DPC Moorepark Fermoy Co. Cork, Ireland mauty-at-dpc.teagasc.ie
I want to study the cell walls changes of surface cells during frying of potato. My idea is:
1st- colour the potato cells at the surface and observe it with a light surface microscope
2nd- fry the potato at 180 celsius
3rd- observe the same cells at the surface without any other treatment
The first step it is easy to do. I used safranin to colour the cell walls. But, when I fry the potato, the dye disappears.
This may sound ridiculus but, does anyone know of a dye or a treatment that may fix the dye to cell wall even after heating to temperatures of 180 celsius degrees ?
I had to look at nano particles of Ag before and I know what you have to deal with. What I found best was to a specimen of sputtered Ag on a carbon support grid to use as a standard. I then proceeded to set up for darkfield conditions using one of the rings produced by the standard. When I went back to the particles they lit up under the darkfield conditions and were easy to locate. You may try something similar with a crushed standard of AgI and CdS. Good luck.
Roberto Garcia EMF Manager Wright State University
I have warn you on the subject line. This is tentative answer for CTF correction in image reconstruction. If you are not interested in this subject, or even you are, this is going to be a long, boring email!
Thank all the people who give me clues or references! After the brief reading of some of those literature, I try to come up with answers to those questions I put up last time. If anything is wrong regarding these answers, it is my fault, not the person who gave me hints!
} What kinds of contrast exists in electron microscopic image } for weak-phase object? Is amplitude contrast the same as } aperture contrast?
For weak-phase object like thin biological specimen, there are basiclly two kinds of contrast: amplitude and phase contrast. Phase contrast is produced by the interference of the scattered electron wave with the unscattered or background electron wave. Amplitude contrast is produced by the loss of electrons which are scattered outside the objective aperture.
} What is the contribution of inelastic scattering on the image } contrast? How to account for it?
During inelastic scattering the incident electron loses energy in the specimen and produces a significant spread of wavelengths, so large in fact that the scattering of these electrons produce essentially a continuous background in the electron diffration pattern.
Zero-loss energy-filtered images of frozen-hydrated specimen have been shown to have higher contrast and improved structural resolution.
} How does partial spatial coherence and temporal coherence } influence the image and electron diffraction?
The coherence of electron source has an important role in the high resolution imaging of weak-phase objects. The partial temporal and spatial coherence both contribute as a multiplicative factors on the CTF, imposing a resolution limit by attenuating high order spatial frequencies. In the focal plane, the partial coherence broadens the diffraction spots.
} Is it always sufficient to use just first order approximation } ( the linear theory of phase and amplitude contrast image } formation ) for CTF consideration?
Yes, it works fine in most cases of biological samples.
} Is it true for low spatial frequencies ignoring the CTF was } better than compensating for phase contrast alone?
In this regime the amplitude contrast is most important. Phase contrast vanishes, and if you correct for this by the inverse of the phase CTF, the error will be considerable.
} I guess compensation for the CTF was necessary and sufficient } to accurately reconstruct molecular densities. Could anyone tell } me the current ways for accurate determination of CTF?
There are basiclly two ways to approach CTF correction. The first one is using diffractogram to obtain the Thon rings, from that to estimate the defocus values or even the ratio of amplitude contrast to phase contrast by the use of two different defocus settings.
Dr. Frank have introdued a second method which accounts for not only the amplitude and phase contrast components, but also all the envelope functions which attenuate the high resolution information and noise. Least-square refinement of the theoretical value against experiment data gives rise to the each parameters in the CTF. They even combine the CTF correction with the 3D reconstruction in a single step to reduce the error and improve resolution of final results.
A recent paper by Ichise described a phase spectra based method in the measurement of TEM parameters ( defocus, astigmatism and beam tilt misalignment ) and the image drift. The phase spectrum is the phase part of the cross-spectrum between two different images due to beam tilts, in which the cross-spectrum is defined as a Fourier transform of the cross-correlation function.
For 2D crystal, Dr. Henderson had set up the ways to accound for phase shift due to tilts and the refinement proticol in astigmatism correction. Spot-scan is supposed to reduce phase gradient during tilts. But for tomograph, I have no clue how to correct phase gradient.
In a recent paper, Dr. Fuller have pointed out the improvement of resolution in icosahedral virus reconstruction attributed not only the number of particles used but also the improved quality of the data from better microscope. And also stressed the importance of CTF correction in the final structure.
} By improving the different components in CTF, what is the optimal } contrast could be achieved in image?
This is mainly limited by the noise. Dr. Brink reported a contrast at 0.42 by spot-scan at 400 KV.
} Is it possible to put EM reconstruction on the same scale with } the structures derived from X-ray crystallography and NMR after } deliberate correction of CTF, solvent effects and differences between } atomic scattering factors for electron and X-ray?
Absolutely.
} To what extend we could use the insights obtained by building } models and comparison of the models with EM reconstruction?
From the limited knowledge of mine, there are several groups trying to gain insights from combining EM data with X-ray structures. Dr.Stewart reported the combining of capsid protein structures from X-ray into EM reconstruction, and discussed several issues on the combination. Dr. Baker have done a lot of work on the virus structural and functional studies based on the results of the known crystal structures and their EM reconstructions. Dr. Milligan et al. derived the mechanism of muscle contraction based on docking the myosin and actin crystal structures into cryo-EM reconstruction of muscle fibers. Dr. Frank combine the crystal structure of tRNA into ribosome 3D reconstruction to deduce the protein translation machanism. There might be tons of other work out there I just have not read yet, please forgive me!
References:
Books:
"Experimental high-resolution electron microscopy", Spence, John C.H., New York : Oxford University Press, 1988. "Transmission Electron Microscopy", Reimer, Ludwig, Berlin ; New York : Springer-Verlag, 1989. "High-resolution transmission electron microscopy and associated techniques" edited by Peter Buseck, John Cowley, and Leroy Eyring. New York : Oxford University Press, 1988.
Journal articles:
Erika J Mancini, Felix de Haas and Stephen D Fuller.(1997) High-resolution icosahedral reconstruction:fulfilling the promise of cryo-electron microscopy. Structure 5 : 741-750.
Zhu J; Penczek PA; Schrvder R; Frank J. (1997) Three-dimensional reconstruction with contrast transfer function correction from energy-filtered cryoelectron micrographs: procedure and application to the 70S Escherichia coli ribosome. J Struct Biol, 118: 197-219
Frank J; Penczek PA. (1995) On the correction of the contrast transfer function in biological electron microscopy. Optik, 98: 125-129
Wade R.H.; Frank J. (1977) Electron microscope transfer functions for partially coherent axial illumination and chromatic defocus spread. Optik, 49:81-92
Ichise N.; Baba N.; Nagashima H. (1997) The phase spectrum-based measurement of the TEM parameters. Ultramicroscopy. 68:181-200
Erickson H.P. & Klug, A. (1970) Measurement and compensation of defocusing and aberrations by Fourier processing of electron micrographs. Phil. Trans. Roy. Soc. Lond. B261, 105-118
Erickson, H.P. (1973) The fourier transform of an electron micrograph - first order and second order theory theory of image formation. In: Advance in optical and electron microscopy, 163-199
Smith, M.F. & Langmore, J.P. (1992) Quantitation of molecular densities by cryo-electron microscopy: Determination of the radial density distribution of tobacco mosaic virus. J. Mol. Biol. 226, 763-774
Schroder, R.R., Hofmann, W. & Metrenet, J.F. (1990) Zero-loss energy filtering as improved imaging mode in cryoelectronmicroscopy of frozen hydrated specimens. J. Struct. Biol. 105, 28-34
Stewart, P. L., S. D. Fuller and R. M. Burnett (1993) Difference imaging of adenovirus: Bridging the resolution gap between x-ray crystallography and electron microscopy. EMBO J. 12:2589-259.
Toyoshima, C. and N. Unwin (1988) Contrast transfer for frozen-hydrated specimens: determination from pairs of defocused images. Ultramicrosc. 25:279-292.
Toyoshima, C., K. Yonekura and H. Sasabe (1993) Contrast transfer for frozen-hydrated specimens II. Amplitude contrast at very low frequencies. Ultramicrosc. 48:165-176.
Brink, J. and W. Chiu (1991) Contrast analysis of cryo-images in N-paraffin recorded at 400kV out to 2.1E resolution. J. Microsc. 161:279-295.
Any comments or corrections will be heartly welcomed! By the way, I have put a copy of this at the end of my cryo-EM reconstruction homepage: " http://www.sb.fsu.edu/~jinghua/em.html "
Yves This may seem a trivial thing but is a source of constant annoyance when I am involved in COSHH (Control of Substances Hazardous to Health) risk assessments because it isn't just water but lots of others including NaCl, PBS, sucrose, glucose. It would be useful if suppliers of these data sheets could preface their hazard information with detail such as low risk unless ingested by the kilogram because sometimes you can very easily come across more hazardous materials with little known information which may seem more innocuous. Some people will then argue, according to the data, that the reagent is no more dangerous than sodium chloride solution.
Malcolm Haswell e.m unit University of Sunderland UK ----------
On Fri, 8 Aug 1997, Ian MacLaren wrote: } } One of the funniest things that I have seen is safety data for water in one } of these books of chemical hazards. It had phrases like: if splashed in } eyes rinse with plenty of WATER, and in case of a spillage wash down drain } with plenty of WATER!!
Ian. here is the copy of that text, sent on this list a few month ago:
Hello all
This is a slightly facetious input re. safety data sheets!
In the UK there is a very useful pair of publications from BDH, a chemicla supply company (used to be known as British Drug House, I believe). These are collcted data sheets. Yes folks, there is one for water. Here are a few salient points which all users of the substance should bear in mind at all times:
1. colourless liquid - maybe that implies it is rare and difficult to see if dropped! 2. Against solubility in water: miscible in all proportions. 3. fire and explosion hazard: not applicable (thats a relief - although if it caught fire I suppose one could foolishly attempt to extinguish with more water?). 4. Health hazard: no significant hazard expected, may be irritating to the eyes. 5. Toxicity: no data. 6. Carcinogenicity: no evidence of carcinogenic properties. 7. First aid - eyes: irrigate thoroughly with water(!) 8. First aid - lungs: remove from exposure. 9. First aid - skin: wash off thoroughly with soap and water (work that one out!). 10. First aid - mouth: wash out thoroughly with water. In severe cases obtain medical attention(!) 11.Reactive hazards: violent reaction with acyl halides, alkali and alkaline earth metals .... 12. Spillage disposal: wear appropriate protective clothing - listed are gloves, goggles, face shield and protective apron. Luckily, respirators are not needed.
We are once again broadcasting Video from the Annual Microscopy & Microanalysis Meeting (this year in Cleveland). Join us virtually on the MSA WWW site
Melanie, I should have mentioned that the 6400F is a field emitter. Thank you.
Jouko, I am sure that charging is not the problem. It even occurred with sputtered gold on carbon resolution samples.
Jan, it sounds as though you were experiencing similar effects. I assume the active field canceling equipment has solved your problem? I would expect the railway to cause the image to shift gradually, peak, and shift back. Not the sudden movement, pause, and sudden return. If related to switching on the motors, I would expect a sudden shift, followed by a gradual return as the train moved off (or the opposite for switching off). JEOL did extensive trouble shooting on my system. They eventually brought in a manufacturer of active field and vibration cancellation equipment as a consultant. My column was placed on an active air table, and inside an active field cancellation system. The conclusion was that the problem was inside the SEM. JEOL eventually exchanged the entire column assembly and the high voltage tank. The difference was like day and night. The resolution and signal to noise were good. The jumping image was gone.
Lately, I have noticed that the jumping image might be returning (hence my inquiry here). I first noticed it at an extraction voltage of 600V (original problem showed up at all voltages). I have seen it at 1kV a couple of times.
JEOL has checked and/or replaced the scintillator a couple of times. The noise canceller has been checked numerous times. The bouncing brightness continues to occur randomly. I have tried flashing the tip when this occurs, and it seems to help. I have not made direct correlation. Perhaps tip instability?
A critical element in imaging the nano-particles is the nature of the support film. Ultra-thin carbon films would be a better bet and you could also try lacey films which would allow you to image the paricles without an underlying substrate.
If you are unable to make these yourself, our company - Ted Pella Inc., is a major manufacturer of support films and we receive many requests for our ultrathin carbon films from researchers studying nano-particles. Please contact us directly if you wish for more information: tedpel-at-aol.com or 1(800) 637-3526.
} I am trying to assist a research group interested in looking at } nanoparticles. Some are made of silver iodide and others are made of } cadmium sulfide, sometimes with a little zinc. They are supposed to be } about 5 micrometers
I got really excited until I saw your correction. I thought that the HVEM would be necessary to see these.
} in size. They do the synthesis and some sorts of } analysis, infrared spectra, I think. They come to my TEM to check on the } size. } } Well, I am having a hard time seeing them.
You could try higher beam current as one responder suggested, but I'd suggest going the other way--use a very sensitive detector and very low beam currents. We have an intensified CCD on our scope which can be used for scanning and focussing with picoamp currents. We also use a small condenser aperture so that no beam is deposited on the sample outside the area being investigated. When the particles have been loca- ted and the image focussed (~1 sec is all that is required), use LoDose film or a slow-scan CCD to record the images. This technique works with TiO2 particles in a similar size range at 1.2 MV (where the contrast is much lower than at 100 kV). Good luck. Yours, Bill Tivol
Mel: You are very correct in letting the microscopy fraternity know about the dangers of dihydrogen monoxide. May I add a further warning that this material which also has another synonym "bis-nor-ethanol" is triphasic and a near universal solvent. Let us be vigilent.
Patrick Echlin
Cambridge, UK also has On Thu, 7 Aug 1997, Melvyn Dickson wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear Colleagues, } } Some eagle-eyed environmentalists have discovered yet another Government } conspiracy to conceal the true harmful nature of a chemical which commonly } contaminates the environment with dire results. } } Full details of this biohazard, dhihydrogen monoxide, can be found at the } following website:- } } http://www.cs.oberlin.edu/students/jbayes/text/dhmo } } If you are properly concerned about the dangers, you may join the coalition } to ban this noxious substance. } } } Mel Dickson } } } } } } } Mel Dickson } Electron Microscope Unit, } University of New South Wales. } Sydney NSW 2052 Australia } } Phone (+612) 9385-2945 } Fax (+612) 9385-1067 } } Website {http://emunit1.babs.unsw.edu.au/emu_top.htm} }
Postdoctoral Research Position in TEM of Semiconductor Epitaxy is Available
Immediately available is a position for a postdoctoral research associate within the Department of Physics at the University of Illinois in Urbana, Illinois, USA. The selected person will study strain distributions in Ge/Si and related epitaxial films by transmission electron microscopy, and will use quantitative measurements of strain to understand the driving forces for island organization and size selection. Both ex-situ microscopy, and in-situ microscopy on our UHV MBE TEM will be involved. The project is supervised by J. Murray Gibson, and involves a collaboration with Jim Coleman, David Cahill and Joe Greene within the UI campus, and the Hewlett Packard Palo Alto Research Laboratory, under NSF funding. The position is for two years (one year at a time, with possible extension to three) and the salary is about $32,000 per year. We are looking for someone with a strong background in conventional diffraction contrast TEM, including the theoretical underpinning and the experimental methods. Experience in semiconductor thin film growth and/or surface science would be an asset. Please communicate by e-mail (preferably) with Murray Gibson at j-gibson-at-uiuc.edu, or call me at 217-333-2997. Note, a second position using our new Low-Energy Electron Microscope and the Scanning Tunneling Microscope on the same project is also available, for which someone with surface science expertize would be most suited.
J. Murray Gibson Professor of Physics and Materials Science Associate Director, Frederick Seitz Materials Research Laboratory University of Illinois, 104 S. Goodwin Ave (Room 258) Urbana, IL 61801 Tel: (217)-333-2997; Fax: (217)-244-2278; j-gibson-at-uiuc.edu
About a year ago I asked for tips on preparation of skin for TEM of stratum corneum. I received some useful replies - Thanks to all!
Now I am looking for references which might show the interaction of medical tapes and dressings with skin, preferably TEM cross sectional images but SEM and LM are interesting as well. So far my literature searches (MEDLINE) have turned up nothing. Any leads would again be appreciated!
Thanks and hope those at MSA are having an enjoyable week!
Karen
-- Karen Zaruba kszaruba-at-mmm.com 3M Company, 3M Center Bldg. 270-1S-01 St. Paul, MN 55144 "The opinions stated above are my own, not necessarily 3M's"
A postdoctoral position at Department of Physics, University of Oslo, is available for a period of two years. The successful candidate will work on a basic research program entitled "Electron microscopy, synchrotron and neutron studies of precipitation and growth in aluminum alloys". The main task will be TEM and/or SANS studies of the early stages of microstructure development in aluminum alloys. Educational requirements include a Ph.D. (or equivalent) in materials science, metallurgy or solid state physics/chemistry and with interest in experimental work. Application deadline is Sept. 15 1997.
For additional information please contact Prof. Tore Amundsen at : tore.amundsen-at-fys.uio.no or Prof. Johan Taft=F8, tel. +47 2285 6113 or +47 2295 8737
A postdoctoral position at Department of Physics, University of Oslo, is available for a period of two years. The successful candidate will work on a basic research program entitled "Electron microscopy, synchrotron and neutron studies of precipitation and growth in aluminum alloys". The main task will be TEM and/or SANS studies of the early stages of microstructure development in aluminum alloys. Educational requirements include a Ph.D. (or equivalent) in materials science, metallurgy or solid state physics/chemistry and with interest in experimental work. Application deadline is Sept. 15 1997.
For additional information please contact Prof. Tore Amundsen at : tore.amundsen-at-fys.uio.no or Prof. Johan Taft=F8, tel. +47 2285 6113 or +47 2295 8737
A postdoctoral position at Department of Physics, University of Oslo, is available for a period of two years. The successful candidate will work on a basic research program entitled "Electron microscopy, synchrotron and neutron studies of precipitation and growth in aluminum alloys". The main task will be TEM and/or SANS studies of the early stages of microstructure development in aluminum alloys. Educational requirements include a Ph.D. (or equivalent) in materials science, metallurgy or solid state physics/chemistry and with interest in experimental work. Application deadline is Sept. 15 1997.
For additional information please contact Prof. Tore Amundsen at : tore.amundsen-at-fys.uio.no or Prof. Johan Taft=F8, tel. +47 2285 6113 or +47 2295 8737
I will be in Cleveland today at the MSA Show. Are there any vendors other than Kevex, BioRad and Evex Analytical that I should visit. I am very eager to see what they have in terms of hardware and software.
Does RuO4 attack glycerol? Also is there a protocol for measuring the concentrations of stained species? I have been working with RuO4 to stain polyvinyl alcohol and need to quantify the concentration / concentration gradient of the polymer. Thanks in advance for any suggestions
Sincerely
Michael Mandanas Particulate Materials Center Pennsylvania State University University Park, PA 16801 mxm67-at-email.psu.edu
Mike, I have only received two responses so far, thanks to both Maria and Lilith, here they are as requested. The first refers to the neurotransmitter GABA.
An additional source I have found for GABAA receptor antibodies is Pharmingen. They carry ABs against alpha 1, 3, 4 and 6. ABs against 3,4 and 6 are suitable only for immunohistochemistry, where anti-alpha 1 is also suitable for western blotting and immunoprecipitation.
On Mon, 4 Aug 1997 16:57:31 -0400 Maria Mejia wrote:
} From: Maria Mejia {maria-at-skivs.ski.org} } Date: Mon, 4 Aug 1997 16:57:31 -0400 } Subject: GABA antibody } To: s.miksys-at-utoronto.ca } }
} Here in our lab. in San Francisco, we are using rabbit } anti-GABA from Sigma cat. # A-2052 with excellent results!! } We did try the GABA from Boehringer Mannheim several times, } but we were never happy with results. Although, we are using } turtle retina tissue - the GABA results are just amazing - } it's a very clean and specific antibody. You have to work } on the right dilution for your application needs, but for us } we use 1:15,000 - overnight -at- RT for 12hrs w/ mild agitation. } We are doing the immuno on frozen sections -at- 10ums thick. } Good luck } maria mejia } smith-kettlewell eye res. inst. } S.F. Ca.
The second response is from Lilith Ohannessian-Barry from the National Research Council Institute of Biological Sciences in Ottawa. The NRC has a very good antibody to alpha 1 subunit, but they have no vendor as yet. If any-one is interested I can put you in touch with Lilith.
I am trying to heat a glass powder which contains an organic binder to= =20 1000 degrees C to observe the binder burnout and glass transition=20 temperatures. However, I need advice on how to prevent the powder=20 from moving during heating. The particles eventually move around,=20 which wouldn't be such a problem except that they often "jump" up onto= =20 the heat shield and obliterate the field of view. I hesitate to=20 adhere the particles to the alumina crucible, since anything I use=20 might react with the binder which is already present in the powder (I=20 do not know what the binder is). =20 Any suggestions would be greatly appreciated. =20 TIA, =20 Leslie Link e-mail: Leslie.Link-at-us.gtc.boc.com
I have been looking (by TEM) at an alloy with mostly Ni, Al,Pt. The alloy was cycled to high and low temperatures several times. The resulting microstructure is highly twinned and it resembles the microstructures seen in martensitic Fe alloys. We have not been able to find other references which would indicate that Ni, Al,Pt alloys undergo martensitic transformations. I would appreciate any suggestions or references that might be useful.
Does anyone have any old Latico "T" series condensor lenses for our Durst Enlarger (SM183) they would be willing to give up? We would pay fair price for any or all of 240T, 200T, 130T and 85T. Thanks!
I would like to announce that the URL of my series of WWW pages called "Microscopy and Imaging Resources on the WWW" has been changed. The University of Arizona College of Pharmacy (which hosts these pages) has decided to retire their old domain name and move to a new one. Currently all page requests for a URL using the old name of "www.pharm.arizona.edu" are being forwarded automatically to "www.pharmacy.arizona.edu", BUT this forwarding will cease in about 6 months.
In preparation for this announcement I have updated the pages to clean out dead links and add new ones I've located. I may eventually stoop to using the dreaded {BLINK} tag near the top of each page to remind visitors to check their links (I promise I'll wait until the end is near).
I appreciate those of you who are the sources of many of the links on my pages. My goal for these pages has been to "collect relevant sources that provide educational and technical information about biological microscopy and imaging in a manner that is non-commercial (in the case of information from vendors) and clear enough for graduate students who are neophytes to microscopy and imaging." If you know of other resources that I'm missing, please contact me.
If you have a reciprocal link to one of my pages I will try to contact you directly to remind you of this change.
Thanks to all who are regular visitors or who have linked to these pages. This project is an outgrowth of my position as manager of the Experimental Pathology Service Core for the National Institutes of Environmental Health Sciences (NIEHS) funded Southwest Environmental Health Sciences Center (SWEHSC). NIEHS is very strong on community outreach and education and I figure these WWW meta-lists fit in rather nicely with that goal.
THE CORRECT URLs ARE:
Microscopy and Imaging Resources on the WWW http://www.pharmacy.Arizona.EDU/centers/tox_center/swehsc/exp_path/m-i_onw3. html
Histology on the WWW http://www.pharmacy.arizona.edu/centers/tox_center/swehsc/exp_path/w3-histo. html
Use & Misuse of Formaldehyde Fixatives
http://www.pharmacy.Arizona.EDU/centers/tox_center/swehsc/exp_path/formalde. html
Other Commonly used Histologic Fixatives
http://www.pharmacy.Arizona.EDU/centers/tox_center/swehsc/exp_path/otherfix. html
Confocal on the WWW http://www.pharmacy.arizona.edu/centers/tox_center/swehsc/exp_path/conf_www. html
Electron Microscopy on the WWW http://www.pharmacy.arizona.edu/centers/tox_center/swehsc/exp_path/em_w3.html
Digital Imaging on the WWW http://www.pharmacy.arizona.edu/centers/tox_center/swehsc/exp_path/dig_imag. html
Free Publications of Interest to Microscopists http://www.pharmacy.Arizona.EDU/centers/tox_center/swehsc/exp_path/freemags. html
Reciprocal Links http://www.pharmacy.Arizona.EDU/centers/tox_center/swehsc/exp_path/m-i_link. html
Reference Material for Biological Scientists http://www.pharmacy.Arizona.EDU/centers/tox_center/swehsc/exp_path/referenc. html
(There are a few other pages, but they are mostly of interest to local folks.)
Yours, Doug Cromey ..................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Sr. Research Specialist 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
Dear Jordi, Seeing something resembling martensite under TEM does not necessarily mean the alloy has undergone martensitic transformation. Actually, it is quite common to see the twinned structure which may not be a result of martensitic transformation in many high temperature alloys such as Ti-Al, Ti-Al-Nb, Ni-Al and so on. Twins may form through phase transformations including martensitic transformation, or mechanically/thermally introduced strains. Your case seems to be most explanable by the later.
There are two most important features of martensitic transformation which I can remember by now. One is the speediness of transformation and the other is a certain crystallographic relationship between parent phase and newly formed martensite.
Chao-Ying Ni Microscopy Division Batta Laboratories, Inc. Newark, DE 19713
On Wed, 13 Aug 1997, Marti, Jordi wrote: } } I have been looking (by TEM) at an alloy with mostly Ni, Al,Pt. The alloy } was cycled to high and low temperatures several times. The resulting } microstructure is highly twinned and it resembles the microstructures seen } in martensitic Fe alloys. We have not been able to find other references } which would indicate that Ni, Al,Pt alloys undergo martensitic } transformations. I would appreciate any suggestions or references that } might be useful. } } Thank you, } } Jordi Marti }
Does anyone have any experience with the SEM/EDS analysis of soot resulting from incomplete combustion? Can differences be detected in soot that results from different fuel sources such as wood, oil burning furnaces, natural gas, LP gas etc.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Does anyone have any experience with the SEM/EDS analysis of soot resulting from incomplete combustion? Can differences be detected in soot that results from different fuel sources such as wood, oil burning furnaces, natural gas, LP gas etc.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
All,
We are getting ready to order the Kevex upgrade to replace our existing IOMEGA 10 Meg drives with the dual Syquest 44 Meg drives on our 9 year old Kevex system.
I was wondering if anyone else had done this and if there were any tips, tricks or warnings with installation or use of these drives?
Thanks for any words of wisdom!!
John Giles Senior Materials Engineer Honeywell Space Systems
Does RuO4 attack glycerol? Also is there a protocol for measuring the concentrations of stained species? I have been working with RuO4 to stain polyvinyl alcohol and need to quantify the concentration / concentration gradient of the polymer. Thanks in advance for any suggestions
Sincerely
Michael Mandanas Particulate Materials Center Pennsylvania State University University Park, PA 16801 mxm67-at-email.psu.edu
Jake, I also have bright horizontal streaks flash across the screen, sometimes. These are caused by emission noise, and a "flash" usually clears them up (unless they are from a charging sample). The problem I was referring to, shows up as "bands" of different levels of brightness (i.e. Start a slow scan; maybe the first 1/2" is one brightness level, the next 1/4" is brighter by several shades of gray, the next 3/4" may be back to the original level of brightness. The widths, and positions, of the bands are random, I only tried to illustrate the problem). The sudden lateral movements of the image are while remaining in super rapid. The shifts when switching from super rapid to slow, and back, are due to changing blocks of circuitry in the console. These shifts can be minimized by your service people.
I've received several private comments about the URLs I posted yesterday. Many have said that the links didn't seem to work. I suspect that in most cases the rather long URLs have "wrapped" from one line to the next. I think you'll find that if you include the part of the URL text that wrapped onto the next line that the links will work (all of them work for me). I'm sorry the URLs are so long, unfortunately the same folks that changed the server name on me won't let me have shorter URLs.
Yours, Doug Cromey ..................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Sr. Research Specialist 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
They kept trying to say that my problems were fields, but the active field cancellation could not affect the visual symptoms. The ultimate proof that it was inside the system, is the fact that the problems went away when they replaced the entire column. It is easy for SEM manufacturers to blame fields for problems they can not trace, or easily solve. They all seem to use this crutch from time to time.
Do not get me wrong, I know fields can be a problem. My response to that is that, as the systems become more and more susceptible to the ocean of magnetic noise they live in, the manufacturers need to prevent the affects with their design, and not use it as an excuse for poor performance in the real world.
Enough of that! I was just wondering if anyone else was having these problems on similar systems. It seems as though there are a few out there.
Greetings to all, I do TEM on human tissues ... mostly renal biopsies. I have noticed that the past 2 or 3 that I have processed have had tiny "pin holes" when viewed under the scope ... don't know why! I have changed NOTHING in my procedure, but feel this is a processing problem. (I hand process everything.) I use Spurr's for embedding ... could one of the four ingredients have gotten contaminated somehow ... maybe with moisture? Should I open all new bottles and try that? Maybe air bubbles are formed when I mix/stir the Spurr's ... that's never happened before when I have mixed it. Has anyone else had this experience just happen "out of the blue" like this? The "pin holes" are so tiny that they don't even show up in cutting the thin sections at 70nm ... only after being stained and put on the 'scope at the END of the whole thing! This doesn't hamper the diagnosis at all, but it's not the quality that I'm used to giving our pathologist and I'm not a bit happy about it! What do you think? Thanks in advance for your help.
Sharron G. Chism HT (ASCP) Electron Microscopy Lab Harris Methodist Hospital Fort Worth, Texas
I am writing from the St. Louis Science Center in St. Louis, MO. The Science Center is a non-profit, hands-on science museum that is free to the public. We have a gallery that is opening next month and we need some good EM and/or light microscopy photographs that show various examples of cell morphology. We would like both animal and plant cell photographs. Needless to say, the photos will have to be of excellent quality, because they will be installed in the gallery and will be on display for the life of the gallery (a minimum of five years). We will be displaying the photographs blown up to at least 8X12 inches as back-lit images on a kiosk.
Does anyone out there have anything to share with us? Please e-mail me directly at cencarna-at-slsc.org. Thank you so much for your help.
Cindy H. Encarnaci=F3n, Ph.D. St. Louis Science Center
A post - doctoral research associate position exists within the interface science group of the Chemistry and Materials Science Directorate of Lawrence Livermore National Laboratory. The group studies in a fundamental research program the relation between the structure and composition of internal interfaces and their properties, including mechanical, diffusional, and mobility. The materials of interest are primarily metals, but interfaces in ceramics and metal/ceramic heterophase interfaces are also studied. The position is for an experimentalist to characterize and quantify grain boundaries and interfaces in regards to their structure, morphology, segregation of minor species, and diffusion. Model interfaces are fabricated for study by diffusion bonding oriented single crystals in our ultra-high vacuum diffusion bonding machine. A primary tool for characterization will be the transmission electron microscope (TEM), but it is anticipated that many other methods could be brought to bear on issues raised by this work. However, conventional and high resolution imaging and analytical techniques will be pushed to their limits by this research. Quantification in the data analysis will require computer code development.
The applicant will be expected to hold a PhD in materials science or a closely related field. A thorough understanding of crystal defects is essential. The applicant will be expected to have extensive experience with TEM. Experience with computer code development, IDL, FORTRAN, and the UNIX operating system is highly desirable. Excellent oral and written English skills are essential. The initial appointment is for one year, with extensions possible for one or more additional years.
Those interested, please contact:
Dr. Geoffrey H. Campbell Lawrence Livermore National Lab Mailstop L-356 P.O. Box 808 Livermore, CA 94551-9900 USA Phone:(510) 423 - 8276 FAX:(510) 424 - 4737 e-mail:ghcampbell-at-llnl.gov
Yes we have had similar "out of the blue" problems with Spurrs on several occasions.
The DMAE component of Spurrs may well have gone bad, it seems to be the most suseptible. I write the date all the bottles when I open them now, in case I am in any doubt. Also check the caps are still good, as I find they occassionally tend to split open in storage.
Another problem we had probably wouldn't apply, as it was caused by a failing seal on our automatic tissue processor: an increase in prop. oxide evaporation during processing resulted in incomplete infiltration.
Hope this helps!
Miss A.J.Wilson Electron Microscope Unit St George's Hospital Medical School Cranmer Terrace Tooting London SW17 ORE Tel: 0181 725 5220 awilson-at-sghms.ac.uk awilson-at-aw.u-net.com
It has been suggested that I should copy this message to all subscribers.
------- Forwarded Message Follows -------
I recently upgraded my Kevex 8000 to use the Syquest drives. Everything went smoothly and the new drive system is working great. The kevex upgrade struck me as somewhat pricy, considering the actual "street price" of the drives, but insures you get the proper cables, support, setup, etc.... One disadvantage is that the 230Mb (for PCs) drive becomes a 44 Mb, DEC/RT11 formatted system.
Eventually, I expect to stop using my 44Mb Bernoullis entirely. To ensure easy availability of recent data during a transition time, my system is currently configured to run one SyQuest drive and the (old) dual 44Mb Bernoullis. Hardware limitations prevented me from running both SyQuests and the pair of 44s.
FWIW... My office PC was setup with an internal 44Ber and an Adaptec 1542 so that I can read the RT11 format data into a DOS/Win system. Using the "spare" SyQuest, I determined that the PC system will support both at the same time.
Woody White Mcdermott Technology, Inc http://www.mtiresearch.com/
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
All,
We are getting ready to order the Kevex upgrade to replace our existing IOMEGA 10 Meg drives with the dual Syquest 44 Meg drives on our 9 year old Kevex system.
I was wondering if anyone else had done this and if there were any tips, tricks or warnings with installation or use of these drives?
Thanks for any words of wisdom!!
John Giles Senior Materials Engineer Honeywell Space Systems
Hi: Since many of the subscribers of the Microscopy mailing list have been asking me about the First Electronic Conference in Analytical Chemistry I have decided to post the following message to explain a little more about it...
************************************** Analytical Chemistry Mailing List ************************************** Due to the success of previous virtual conferences (MGM EC1 & EC2, first and second electronic molecular graphics and modelling conference http://bellatrix.pcl.ox.ac.uk/egc/ and http://bellatrix.pcl.ox.ac.uk/egc2/home.html, ECHET96, electronic conference on heterocyclic chemistry, http://www.ch.ic.ac.uk/ectoc/ehet96/, ECCC1 & ECCC2, first and second electronic computational chemistry conference http://hackberry.chem.niu.edu:70/0/ECCC/homepage.html) it was decided to set up a moderated mailing list to pave the way for the
FIRST ELECTRONIC ANALYTICAL CHEMISTRY CONFERENCE to be held in November of this year (3rd-14th)
Preperation for the electronic conference is still in its early stages and any input (discussion topics, suggested conveners, short courses literature/product reviews etc.) would be greatly appreciated.
You can add your name to this mailing list by sending an e-mail to
ac-request-at-vei.co.uk
leaving the subject line blank and
subscribe {your_email_address}
as the message body (no signatures). Once accepted you will be sent a mail welcoming you to the mailing list and explaining how to post a message.
Among some of the areas we hope to include in discussion will be topical aspects of: * NMR, IR, UV, HPLC and GC. * Electrochemistry. * AA, ICP and ICPMS. * Macromolecular and Small molecule Crystallography. * Analytical techniques. * Method development and validation procedures. * Bioanalytical forum. * MS and applied topics. * Capillary Chromatography & Electrophoresis. * Statistical analysis of data. * Microscopy
Any suggested additions may be made through the mailing list above.
Authors can opt to have their presentation in the following categories:
- non-permanent WWW presentation of a conference poster - non-permanent WWW presentation of a conference lecture - refereed WWW presentation which will be considered for print publication as a full paper in Critical Reviews in Analytical Chemistry.
- refereed WWW presentation which will be considered for permanent electronic publication in the Internet Journal of Chemistry (IJC), http://www.ijc.com/
During the conference interaction, presentations and discussions will take place via the Internet using a Java-based virtual conference centre, WWW-based discussion forums and an electronic mailing list. Before the conference, a timetable for lectures and discussion sessions for each section will be posted. Since these realtime discussions are an integral part of the conference, authors will be expected to attend one for their subject; the right is reserved not to referee submissions by authors who do not attend one of these sessions.
The Conference will feature a Virtual Exhibition where exhibitors will be able to describe the activities of their organization, display their products and services and interact with registrants. Potential exhibitors should contact the conference organiser.
Here are some basic suggestions for this king of problem: It could be water in you solutions - try new chemicals, but don't throw the old ones away until you're sure they were the problem. OR it could be a change in the size of the tissue? or the fixation process, ie the fix is old. Maybe try putting the Spurrs under a vacuum before you use it, or put the tissue and Spurrs under a SLIGHT vacuum before you embed it. Do you use propylene oxide to dehydrate? I have had some old prop. go bad on me and make all the blocks soft. If it had a small amount of water in it, it might cause your problem. Good luck.
Karen Pawlowski Sr. Research Assoc. ENT Res. Lab., UT Southwestern Medical Center PhD student, UT Dallas Dallas, TX
On Thu, 14 Aug 1997, Chism, Sharron wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Greetings to all, } I do TEM on human tissues ... mostly renal biopsies. I have noticed } that the past 2 or 3 that I have processed have had tiny "pin holes" } when viewed under the scope ... don't know why! I have changed NOTHING } in my procedure, but feel this is a processing problem. (I hand process } everything.) I use Spurr's for embedding ... could one of the four } ingredients have gotten contaminated somehow ... maybe with moisture? } Should I open all new bottles and try that? Maybe air bubbles are } formed when I mix/stir the Spurr's ... that's never happened before when } I have mixed it. Has anyone else had this experience just happen "out } of the blue" like this? The "pin holes" are so tiny that they don't } even show up in cutting the thin sections at 70nm ... only after being } stained and put on the 'scope at the END of the whole thing! This } doesn't hamper the diagnosis at all, but it's not the quality that I'm } used to giving our pathologist and I'm not a bit happy about it! What } do you think? } Thanks in advance for your help. } } Sharron G. Chism HT (ASCP) } Electron Microscopy Lab } Harris Methodist Hospital } Fort Worth, Texas }
If Spurr's resin has served you well in the past, I stick (sorry) with it. I'd recommend you get a fresh kit from any of the suppliers out there who handle the stuff. In our lab, where the humidity is condensing from May until snowfall, we have at least one bout of bad resin nearly every summer. My own suspect is that the anhydride winds up not so after a few people leave the lid loose, either on the stock bottle, or on an individual preparation. We also refrigerate our mixtures to extend the pot life and despite my warnings I'll bet there are those who don't let them reach the dew point (i.e. RT) before opening. Given the relatively low price of the kits, I'd reckon it's worth one's time to get a fresh one.
With respect to mixing resins, Spurr's is an epoxy mix to begin with, but if you don't like the characteristics of the published formulas, you can cook up your own. I've taken to using Quetol 651 in an equiequivalent mix with the VCD instead of the DER resin. We call it Spurtol. It seems to stain a little easier than the DER softened mix. DMAE works as an accelerator just as in A.Spurr's original. Works well with plants, chicken, and some high density polymers.
cheers, John Heckman TEM Supervisor/ Academic Specialist MSU Center for Electron Optics Michigan State University
Hi Folks, A little good news for those of us unwilling/unable to pay $6 billion for a RIP to network the Fujix printer. A company called Techpool software in Cleveland (www.techpool.com) have produced a RIP which costs $490 and seems to work pretty darn well. This allows printing from any application or across the network. The way the networked system works is like the Lasergraphics sildemaker, you mount a "hot" or watched folder and save printjobs to that folder and a TSR on the computer with the printer watches that queue for jobs. Works pretty well in our hands (though we are still using the demo, which you can get from the web site)
BTW I have no interest in this company etc etc Simon
-- Simon C. Watkins Ph.D. Associate Professor Director CBI University of Pittsburgh Pittsburgh PA 15261 tel:412-648-3051 Fax:412-648-2004 URL:http://sbic6.sbic.pitt.edu
Do any of you have experience labeling cell surface antigens for SEM? I've been working with someone who has a couple of these projects in mind. We made one attempt at gold labeling in the standard fashion but were thwarted by the low resolution of my non vibration-mounted scope. Or maybe the labeling didn't work in the first place.
I think this person will have to go elsewhere to get what she needs because of the limitations of my scope but I told her I'd post a request for established protocols and any tips you might have to offer. The particular experiment she has in mind is to visualize annexin 4 on the surface of HUVEC cells.
Thanks in advance for your help, Tori
Tori Hatch thatch-at-hsph.harvard.edu Physiology Program Harvard School of Public Health 665 Huntington Ave. Boston MA 02115
Just so that every one knows Taab 812 is ~6 times more expensive than the conventional spurr or epon... Neelima Shah
At 12:58 PM 8/15/97 GMT+0200, ROBIN CROSS wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The following is posted as a favor to one of the companies which represents our products.
Pulcir, Inc. has an opening for a sales/support engineer in the Southeastern U.S. with experience in electron microscopy and EDS. If interested please contact:
A six month maternity-cover postdoc. position is available in my group from 1.10.97. Applicants require some knowledge of cell culture and electron microscopy to investigate the interaction between living tissues and new glass-ceramic biomaterials with dental and orthopaedic uses.
Further information and application details are available from:
The Director of Human Resource Management University of Sheffield Western Bank SHEFFIELD S10 2TN email: Jobs-at-sheffield.ac.uk
Quoting refererence R1286
I am happy to deal with informal questions.
Dr Paul V. Hatton Lecturer in Biomaterials School of Clinical Dentistry University of Sheffield Claremont Crescent SHEFFIELD S10 2TA
Tel. (0114) 271 7938 Fax. (0114) 2665326 or 2797050
It has been pointed out to me, and rightfully so, in my previous posting about the price of Taab as compared to Epon or Spurr I neglected to state that the cost in USA. I have no personal or financial interest any one of the products but since in recent times cost has become a major concern in most institutions i thought it necessay to point out. I apologize for my haste in posting. Neelima Shah Regards... :-) :-) :-) :-) :-) :-) :-) :-) :-) :-) ;-) ;-) Visit us at http://www.med.upenn.edu/~path/core/EMCMAIN1.htm
MSA is collaborating with the Lawrence Hall of Science, an outstainding science education center, to publish a classroom manual as part of Project MICRO, MSA's middle school educational outreach program. It's titled "Microscopic Explorations", and publication is scheduled for May '98. LHS materials are used in 25% of U.S. schools, so it will get wide distribution. The LHS has asked for color micrographs to use on all four covers (inside & outside, front & back). We need LM more than EM, at fairly low magnifications, and we need them by SEPTEMBER 25. Topics presented in the manual are preferred: pond life, brine shrimp, insects, crystals of common substances, sand grains, fabrics, color printing - but other eyecatching things that are familiar to children are welcome. Your contribution will be acknowledged in the manual. Please let me know directly, ASAP, if you are interested in sending an image.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
this is not my field but I do know of someone who had problems with gold labelling on the SEM.
You haven't given much information: 1.The name of your microscope and resolution 2. The size of gold particle (eg if you are using 5nm gold they are extremely difficult to see against a thick SEM sample not just because of resolution but because of insufficient thickness to scatter sufficient electrons). If you must use small gold I believe that silver enhancement might be possible. 3. I assume you have only used carbon to coat the samples because most metal coatings will make the gold invisible or difficult to see. 4. Do you have a backscattered detector? Because they are much better at picking up the mass difference of the gold against the lighter elements of the specimen. 5. You could check to see if you can detect the gold on its own by drying some onto the stub - this would at least tell you if it's a resolution problem. If this works you could try to find a positive control and look for the label on that.
If you have done all of the above then I would wait for a real expert to answer.
I hope this helps - but as I said I have never used gold labelling on the SEM.
Malcolm Haswell e.m. unit University of Sunderland UK
Does RuO4 attack glycerol? Also is there a protocol for measuring the concentrations of stained species? I have been working with RuO4 to stain polyvinyl alcohol and need to quantify the concentration / concentration gradient of the polymer. Thanks in advance for any suggestions
Sincerely
Michael Mandanas Particulate Materials Center Pennsylvania State University University Park, PA 16801 mxm67-at-email.psu.edu
For those of you who read the American Lab coverage on Microscopy & Microanalysis '97 and were confused about the source of the book "Optimizing Light Microscopy for Biological and Clinical Laboratories": The book is not available through MSA but is available through MME (Microscopy/Microscopy Education). If you are interested in ordering, please send your Fax number to: Barbara Foster MME (413)746-6931 Fax: (413)746-9311 email: mme-at-map.com We will fax an order form to you, and will honor the 20% M&M '97 discount through Labor Day.
Our apologies for the editorial mix up in both email and annotation.
Barbara Foster Consulting editor to American Lab Consortium President, Microscopy/Microscopy Education
A quick "THANKS!!!" to the nearly 30 companies who contributed to the article covering the Microscopy & Microanalysis '97 meeting. American Lab gave us a whopping 8 page coverage! Several attendees stopped by the MME booth to say that they never knew the meeting existed before the article appeared and several members of MSA council expressed their great appreciation at having such wonderful promotion. For those of you who have not yet seen the article, look for American Laboratory, July, 1997, pp. 38-46.
A reminder that this article was actually part of an on-going column called "Focus on Microscopy" which appears approximately bimonthly in Am. Lab. Contributions are always welcome. Our next column will be on managing images on both Intra and Internet. Please send suggestions and helpful hints to: Barbara Foster Microscopy/Marketing & Education 53 Eton Street Springfield, MA 01108 Ph: (413)746-6931 Fax: (413)746-9311 email: mme-at-map.com
Thanks again! Barbara Foster Consortium President, MME Consulting editor, American Lab
You should first ask how the larger disks will be handled. RT-11 and TSX+ were limited to volume sizes of 32 MB under the DU driver. There was an enhanced DU driver produced by a third party that would handle larger drives as a single volume.
When we added a hard drive to our Delta, we limited it to 170 MB because we were keeping 2 Bernoulli drives which would leave only 6 of the 8 possible DU units of up to 32 MB each to use for the hard drive. Thus anything more than 192 MB would have been wasted space.
There may also be a controller issue. The original Bernoullis on our Delta used a SCSI controller that required the DL driver. That driver supported up to four 10 MB devices. Your new disks will probably require a switch to a DU-type controller (see above) like we had to purchase for our upgrade. Such cards should be available on the used market now at a decent price.
Feel free to ask for more details.
At 08:33 AM 8/14/97 -0500, you wrote: } All, } } We are getting ready to order the Kevex upgrade to replace our existing IOMEGA } 10 Meg drives with the dual Syquest 44 Meg drives on our 9 year old Kevex } system. } } I was wondering if anyone else had done this and if there were any tips, tricks } or warnings with installation or use of these drives? } } Thanks for any words of wisdom!! } } John Giles } Senior Materials Engineer } Honeywell Space Systems ---------------------------------------------------- Warren E. Straszheim 270 Metals Development, Ames Lab/ISU, Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-3091
NIH Image for Macintosh may work for your application. It does have video image capture capability. This is a public domain piece of software. The following is an excerpt from an e-mail about 3-d reconstruction and has the URL from which to download the MAC and Win95 versions of this software.
NIH-IMAGE This popular freeware program for image analyses is originally written for the Mac, but now is also available as Win95 program. Download: http://www.zippy.nimh.nih.gov/ (Mac) or http://www.scioncorp.com/ (Win95) Many information e.g. online manual, macros, example-files and additional download possibilities can be found at: http://rsb.info.nih.gov/nih-image/ As an example animated reconstructions of plant cells (based on semithin
sections) can be found on the home page of Gary Chinga: http://www.nvg.unit.no/~gary Information about the NIH-Image mailing list can be found at http://www.soils.agri.umn.edu/infoserv/lists/nih-image/
Good Luck
============================ Michael D. Standing Electron Microscopy Technologist Brigham Young University ============================
mauty-at-DAIRY.TEAGASC.IE wrote:
} ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } ----------------------------- } -----------------------------------------. } } I have a Matrox Meteor imaging board and would like to acquire RGB } images via a JVC TK1070E CCD camera fitted to a Zeiss confocal } microscope. Does anyone know of any shareware (or fairly cheap) image } acquisition software that I could use to view and acquire images, } preferably as TIFF image ? } } Regards } } Mark Auty } DPC Moorepark } Fermoy } Co. Cork, Ireland } mauty-at-dpc.teagasc.ie
Using the programs named RTDIR.EXE and RTCOPY.EXE (which were a portion of the old Kevex "Report Manager" software package) I can read a 44 Mb Bernoulli or a SyQuest (RT format) disk into the PC clone. The two programs operate much like DOS's DIR and COPY commands. An Adaptec 1542 SCSI controller is used to interface the two drives to the PB bus.
Given this... I can transfer files ok, but the EDS spectra are packed in "library" files and I haven't tried to disect one. It has been a while, but I think you can SAVE/EXTERNAL a spectrum which would (like maps,images etc.) be a discrete file.
Woody White Mcdermott Technology, Inc.
-------------------------------------
Woody,
I appreciate the reply, and have a question for you about your office PC. It sounds like you are able to import the Kevex data into a PC and make use of it. My technician was very interested in your reply and wondered what you are importing with the RT11 data. Specifically, are you able to read the RT-11 data from an individual spectrum on your PC? We have a PC linked to our Kevex system to import Feature analysis data, but it would be nice to be able to import the spectra to the PC also so you could use in a report.
I recently upgraded my Kevex 8000 to use the Syquest drives. Everything went smoothly and the new drive system is working great. The kevex upgrade struck me as somewhat pricy, considering the actual "street price" of the drives, but insures you get the proper cables, support, setup, etc.... One disadvantage is that the 230Mb (for PCs) drive becomes a 44 Mb, DEC/RT11 formatted system.
Eventually, I expect to stop using my 44Mb Bernoullis entirely. To ensure easy availability of recent data during a transition time, my system is currently configured to run one SyQuest drive and the (old) dual 44Mb Bernoullis. Hardware limitations prevented me from running both SyQuests and the pair of 44s.
FWIW... My office PC was setup with an internal 44Ber and an Adaptec 1542 so that I can read the RT11 format data into a DOS/Win system. Using the "spare" SyQuest, I determined that the PC system will support both at the same time.
Woody White Mcdermott Technology, Inc http://www.mtiresearch.com/
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
All,
We are getting ready to order the Kevex upgrade to replace our existing IOMEGA 10 Meg drives with the dual Syquest 44 Meg drives on our 9 year old Kevex system.
I was wondering if anyone else had done this and if there were any tips, tricks or warnings with installation or use of these drives?
Thanks for any words of wisdom!!
John Giles Senior Materials Engineer Honeywell Space Systems
------------------ RFC822 Header Follows ------------------ Received: by smtpmm.space.honeywell.com with SMTP;15 Aug 1997 08:54:30 -0500
Received: from sparc5.microscopy.com by fl51mail.space.honeywell.com with SMTP (1.38.193.5/16.2) id AA00392; Fri, 15 Aug 1997 08:44:37 -0400 Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id HAA02214 for dist-Microscopy; Fri, 15 Aug 1997 07:08:55 -0500 Received: from gate.mcdermott.com (GATE.MCDERMOTT.COM [131.184.96.33]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id HAA02211 for {microscopy-at-sparc5.microscopy.com} ; Fri, 15 Aug 1997 07:08:54 -0500 id AA06559; Fri, 15 Aug 97 07:29:11 CDT Received: by mta.mcdermott.com (1.37.109.20/16.2-WT4.0) id AA187447346; Fri, 15 Aug 1997 07:15:46 -0500 Received: by MCI; Fri, 15 Aug 1997 8:17:00 -0500
mark_munro-at-bio-rad.com wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Does anyone know of any good sources of information on the emission } and absorption spectra of different fluorochromes, and their binding } properties? } } Thanks in advance, } } Mark Munro.Mark,
An excellent reference is the "Handbook of Fluorescent Probes and Research Chemicals" First copy usually free; subsquent ones, nominal cost. Contact: Molecular Probes, Inc. P O Box 22010 Eugene, OR 97402 Ph: (503)465-8353 Fax: (503)344-6504
Best of luck... and if you need further info on fluorescence, please give us a call or email.
Barbara Foster Consortium President Microscopy/Marketing & Education 53 Eton Street Springfield, MA 01108 Ph: (413)746-6931 Fax: (413)746-9311 email: mme-at-map.com
Thank you to all who responded to my TEM artifact problem regarding "pinholes" in the specimen. The suggestions were very helpful and I think the culprit is the bottle of absolute ETOH I have been using. I opened a new bottle today and am processing a renal case. If this doesn't do it, I'll go to the next suggestion and use new Spurr's components. (Someone asked if the biopsies were put in those blue sponges that also cause holes in tissue ... no, the biopsies I get are put straight into small bottles of fixative ... without sponges or cassettes.) Hopefully, the new bottle of ETOH will solve the problem. Thanks, again for your thoughts ... Sharron G. Chism HT (ASCP) Electron Microscopy Lab Harris Methodist Hospital Fort Worth, Texas
Mark - try Molecular Probes, Inc. http://www.probes.com (514)456-8353 they know about fluorochromes! -Mike
On Mon, 18 Aug 1997 mark_munro-at-bio-rad.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Does anyone know of any good sources of information on the emission } and absorption spectra of different fluorochromes, and their binding } properties? } } Thanks in advance, } } Mark Munro. } } }
The following is posted for your consideration. Bob Craig OSRAM SYLVANIA Products Inc.
Scanning Electron Microscopy/X-ray Microanalysis Specialist
Exceptional opportunity exists for a qualified individual to apply her/his analytical and experimental skills in support of research, development, and manufacture of sophisticated lighting products.
This position is responsible for the application of scanning electron microscopy and X-ray microanalysis to study materials problems associated with the development and manufacture of state-of-the-art lighting products. Responsibilities will include: developing new methods for evaluating lamp materials, performing in-depth studies to solve complex lamp materials problems, and operating and maintaining associated analytical instrumentation. The successful candidate will be expected to function as part of a highly skilled technical team, interact in a consultatory fashion with internal R&D and manufacturing customers, report results and make technical recommendations based on the interpretation of those results.
Candidates with the following educational background will be considered: a M.S. degree (research) with 3-5 years experience or a recent Ph.D. in chemistry, materials science or physics. Hands on experience in scanning electron microscopy and X-ray microanalysis of inorganic materials, particularly metals and ceramics, is an absolute must. In addition, the successful candidate should have a firm knowledge of electron-solid interactions and a thorough understanding of the physics of X-ray generation and interaction with crystalline solids. Experience in X-ray diffraction and thermal analytical techniques is also desirable. Good inter-personal, and writing skills are requisite.
Please respond with a resum=E9 to: Mr. Amando Llorente, Human Resource Manager OSRAM SYLVANIA Products Inc. Lighting Research Center 71 Cherry Hill Drive Beverly, MA 01915
I was asked to post the following position announcement by a collegue. The institution would like to fill it for the upcoming academic year.
Heather Owen
Heather A. Owen, Director Electron Microscope Laboratory Department of Biological Sciences University of Wisconsin - Milwaukee (414)229-6816
Full-time Assistant Professor of Biology Academic Year 1997-98
Teaching responsibilities to include:
Introductory Biology
and
Human Anatomy & Physiology
and/or
Integrated Physical/ Biological Science
and/or
Introductory Electron Microscopy
and/or
Histology
Teaching load approximately 12 credit hours per semester. Master's degree required; Ph.D. preferred. New position; may be renewed pending funding. Send letter of interest, curriculum vitae, transcripts and list of references to:
John F. May, Ph.D., Coordinator Department of Biology Marian College 45 South National Ave. Fond du Lac, WI 54935
To those of you trying to respond to our earlier message of thanks for the American Lab article: We are having trouble with our email. Please do not use the "RE:" method of responding. Email directly to: mme-at-map.com.
To those of you trying to get through to Microscopy/Microscopy Education about this book: We are having email problems. Please do not respond using the "RE" function. Send email to: mme-at-map.com.
The techniques listed on the board almost to a person refer to dehydrating the specimen after removing the wax with xylene and then processing as usual. It should be noted that several of my colleagues as well as myself have processed the tissue without rehydrating and then dehydrating the tissue again. The key question is WHAT are you possibly gaining b;y rehydrating the tissue????? The only possible reason or certainly the main one would be to try and "osmicate" the tissue. It has been our observation that the tissue simply does not turn black or pick up any significant amoun of osmium whatsoever after this rehydration. I think that this "old technique" to recover paraffinized tissue is based as much on "urban legend" as anything. The least amount of processing the better. Uranyl Acetate is soluble in absolute acetone for example although en bloc staining of tissue here seems mostly futile. Better to process the tissue directly into plastic after xylene by washing in acetone and propylene oxide and infiltrating into plastic. Afterwords use heavy duty uranium and lead stain ( also I guess one could expose the grids to Osmium vapors to attempt to "osmicate" but I suspect the effort might not be worth it. I would apprecitate some serious comment on this including wheter you have tried this more direct approach. May you never have to do this as I and others have had to for the last 15 years. bob
I am trying to measure the thickness of corneas. Currently, I am using a pachymeter(SD=10 microns). The method is not accurate because it imply compression of the tissue. Could I expect a lower standard deviation from measurements obtained from x-sections using LM? Is anyone familiar with a method that would not involve LM (tissues would not be processed for LM a suitable alternative is employed)? I read a paper where the authors used the universal measuring microscope on corneas embedded in epon. What does that mean? What is is? Could it be good for my purpose?
Thanks in advance,
Dan Caruso Biological Technician medjet-at-worldnet.att
Mark, go into the links on our site. Use search to find 'fluoro' (there are over 400 links) and this will take you to a link at the uni of Buffalo with an extensive table. Just what you are looking for. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 400+ Links, MSDS ************************ http://www.proscitech.com.au ____________________________________ } Does anyone know of any good sources of information on the emission } and absorption spectra of different fluorochromes, and their binding
} properties? } } Thanks in advance, } } Mark Munro. } }
} } It has been pointed out to me, and rightfully so, in my previous posting } about the price of Taab as compared to Epon or Spurr I neglected to state } that the cost in USA. I have no personal or financial interest any one of } the products but since in recent times cost has become a major concern in } most institutions i thought it necessay to point out. I apologize for my } haste in posting. } Neelima Shah } Regards... } :-) :-) :-) :-) :-) :-) :-) :-) :-) :-) ;-) ;-) } Visit us at http://www.med.upenn.edu/~path/core/EMCMAIN1.htm } } Regards... :-) :-) :-) :-) :-) :-) :-) :-) :-) :-) ;-) ;-) Visit us at http://www.med.upenn.edu/~path/core/EMCMAIN1.htm
Friends, We have been notified that our histology service lab here at the University of Arizona may be audited. Because I "inherited" the pricing structure from someone else, the documentation to justify our prices is somewhat lacking. I'm working on that now, but in the meantime would someone be willing to compare notes with me that's gone through such a process?
We would particularly be interested in corresponding with a lab that serves a similar constituency to ours. We serve only University affiliated researchers (we're in the medical college). We do a wide variety of research specimens (mostly the usual rodent tissues, but occasional insects & botanicals), but no clinical work. For more on what we, do our Web site is: http://www.cba.arizona.edu/histology-lab.html
If you can assist me or refer me to someone who can assist me I'd greatly appreciate it. Since this issue may not be of interest to most of the subscribers to this list, please reply to me privately. Thank you.
Yours, Doug Cromey Supervisor, Cell Biology & Anatomy Histology Service Core Lab ..................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Sr. Research Specialist 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
Friends, We have been notified that our histology service lab here at the University of Arizona may be audited. Because I "inherited" the pricing structure from someone else, the documentation to justify our prices is somewhat lacking. I'm working on that now, but in the meantime would someone be willing to compare notes with me that's gone through such a process?
We would particularly be interested in corresponding with a lab that serves a similar constituency to ours. We serve only University affiliated researchers (we're in the medical college). We do a wide variety of research specimens (mostly the usual rodent tissues, but occasional insects & botanicals), but no clinical work. For more on what we, do our Web site is: http://www.cba.arizona.edu/histology-lab.html
If you can assist me or refer me to someone who can assist me I'd greatly appreciate it. Since this issue may not be of interest to most of the subscribers to this list, please reply to me privately. Thank you.
Yours, Doug Cromey Supervisor, Cell Biology & Anatomy Histology Service Core Lab ..................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Sr. Research Specialist 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
Digital Instruments (DI) will be hosting two workshops on Scanning Probe Microscopy in the New England area during the month of September, 1997. The workshops will consist of two lectures in the morning, and a hands-on demonstration in the afternoon. Samples of interest to attendees may be analysed at this time. Those interested in attending these free workshops can get more information at the DI web site (www.di.com) or can contact Rich Goodheart at 410-437-1805.
Eric wrote: ========================================== To the wealth of knowledge out there on the list.. Does anyone know if the Zerostat guns are still available anywhere?? ========================================== They ARE still available, not just from SPI but also from several of our friendly competitors. Remember, Zerostat (R) is a registered trade name and should be indicated as such.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
The Zerostat guns have been discontinued for a few years now however we have now reintroduced them into our line. Please contact us for further information. Stacie Kirsch Electron Microscopy Sciences Tel: 215-646-1566
A full-time microscopy technician position is available in the laboratory of Dr. Ann Hubbard at The Johns Hopkins School of Medicine. The position involves biological specimen preparation, imaging, and image analysis at the light and electron microscopic level. The research environment in the schools of Medicine and Arts and Sciences at Hopkins encourages a cooperative approach that is reflected in collaborations among diverse labs, faculty, students, post-doctoral fellows, and technicians. Thus, there is a potential for interactions with technical and academic personnel throughout the institution.
Qualifications
The successful applicant will have a BS/BA in a biological science and at least 2 years of experience in a wide variety of microscopy techniques. Proficiency in fixation and embedding, ultramicrotomy, immunocytochemistry, and digital and photographic imaging techniques is essential. Experience in the operation of TEMs and the operation and maintenance of research-grade light microscopes is required. Preference will be given to candidates with additional experience in cryoultramicrotomy, confocal microscopy, and computer-aided image analysis.
Duties
All aspects of specimen preparation for a variety of biological samples. Immunofluorescence and immunogold cytochemistry. Imaging and analysis of specimens using advanced light (including confocal) and electron microscopes. Preparation of micrographs for publication. Routine maintenance of light microscopes, an ultramicrotome, and all microscopy related equipment and supplies. Aspects of experimental design and method development as commensurate with experience. Train, advise, and assist other lab members with regard to specimen preparation, microscopy, and imaging. Salary will be in the range of $ 25 - 30 K per annum, or higher, depending on experience.
Send your c.v. and the names and addresses of 3 references to:
Dr. Ann Hubbard Dept. of Cell Biology and Anatomy Johns Hopkins School of Medicine 725 N. Wolfe St. Baltimore, MD 21205
Dear Microscopists: Our imaging centre would like to acquire a 2-photon "confocal" microscope. I know of the Bio-Rad multi-photon system. Have any other companies begun to manufacture and market such systems?
David P. Bazett-Jones, Ph.D.
Professor Departments of Anatomy and Medical Biochemistry Director, Microscopy and Imaging Facility The University of Calgary 3330 Hospital Dr Calgary, AB T2N 4N1 Canada TEL: 403 220-3025, FAX: 403 270-0737
I need desplastificate slices of 4-5 um of insects legs, which are embedded in Durcupan. I have tryed with methoxide of Sodium, and saturated solution of Ethoxide of sodium ( from 1h. to 24 hrs.), and I couldn't. I have problems with the Ethoxide solution too, it become brown after two days. Somebody knows wath can I do?. Thanks, Veronica. Veronica Campanucci -------------------------------------------------------------------------------- Veronica Andrea Campanucci Laboratorio de Fisiologia de Insectos Dpto. de Ciencias Biologicas Facultad de Ciencias Exactas y Naturales Tel (54 1) 781-5021 to 29, ext. 332 Universidad de Buenos Aires FAX (54 1) 782-0582/544-7893 (1428) Buenos Aires e-mail: veronica-at-bg.fcen.uba.ar Argentina HTTP://biolo.bg.fcen.uba.ar/physinse.htm --------------------------------------------------------------------------------
I need desplastificate slices of 4-5 um of insects legs, which are embedded in Durcupan. I have tryed with methoxide of Sodium, and saturated solution of Ethoxide of sodium ( from 1h. to 24 hrs.), and I couldn't. I have problems with the Ethoxide solution too, it become brown after two days. Somebody knows wath can I do?. Thanks, Veronica. Veronica Campanucci -------------------------------------------------------------------------------- Veronica Andrea Campanucci Laboratorio de Fisiologia de Insectos Dpto. de Ciencias Biologicas Facultad de Ciencias Exactas y Naturales Tel (54 1) 781-5021 to 29, ext. 332 Universidad de Buenos Aires FAX (54 1) 782-0582/544-7893 (1428) Buenos Aires e-mail: veronica-at-bg.fcen.uba.ar Argentina HTTP://biolo.bg.fcen.uba.ar/physinse.htm --------------------------------------------------------------------------------
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Reply to: RE} 2-photon microscopy
Dear David, I believe Zeiss is also working on a 2-photon system.
Linda Chicoine Center for Cell Imaging http://info.med.yale.edu/cellimg Cell Biology Yale University New Haven, CT 06520 203-785-3646
--------------------------------------
Dear Microscopists: Our imaging centre would like to acquire a 2-photon "confocal" microscope. I know of the Bio-Rad multi-photon system. Have any other companies begun to manufacture and market such systems?
David P. Bazett-Jones, Ph.D.
Professor Departments of Anatomy and Medical Biochemistry Director, Microscopy and Imaging Facility The University of Calgary 3330 Hospital Dr Calgary, AB T2N 4N1 Canada TEL: 403 220-3025, FAX: 403 270-0737
------------------ RFC822 Header Follows ------------------ Received: by QuickMail.Yale.edu with SMTP;19 Aug 1997 11:44:15 -0400 Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id JAA12038 for dist-Microscopy; Tue, 19 Aug 1997 09:57:04 -0500 Received: from acs7.acs.ucalgary.ca (acs7.acs.ucalgary.ca [136.159.34.207]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id JAA12035 for {microscopy-at-Sparc5.Microscopy.Com} ; Tue, 19 Aug 1997 09:57:03 -0500 Received: by acs2.acs.ucalgary.ca (AIX 4.1/UCB 5.64/4.03) id AA75360; Tue, 19 Aug 1997 09:01:31 -0600 Message-Id: {9708191501.AA75360-at-acs2.acs.ucalgary.ca}
Dennis, Do you embed in Durcupan? Thanks, Veronica. Veronica Campanucci -------------------------------------------------------------------------------- Veronica Andrea Campanucci Laboratorio de Fisiologia de Insectos Dpto. de Ciencias Biologicas Facultad de Ciencias Exactas y Naturales Tel (54 1) 781-5021 to 29, ext. 332 Universidad de Buenos Aires FAX (54 1) 782-0582/544-7893 (1428) Buenos Aires e-mail: veronica-at-bg.fcen.uba.ar Argentina HTTP://biolo.bg.fcen.uba.ar/physinse.htm --------------------------------------------------------------------------------
Hi all, A post-doc has presented me with an interesting task and I would appreciate some help! I have been given some mouse femurs (undecalcified) which were placed in a 1:1 mixture of JB-4 resin (both components A and B in the appropriate mix) and water. The samples were then placed in a vacuum at 20*C and left over the weekend with the aim of enhancing infiltration. On Monday the samples were sitting in a gelatinised resin yuk!! My task is to re-embed them. I am currently trying to dissolve the resin out with many changes of distilled water and constant movement but all that has happened is that the resin has turned white. Any advice would be welcome as these are "valuable" (aren't they all!) samples.
I need to locate any published papers on ESEM applications of petroleum technology and exploration and production in general. Any information will be greatly appreciated.
Sara, You are, as they say in the UK "in a bit of a sticky wicket". JB4 is of course GMA which in it's polymerized state is not soluable in any chemical solution that I know of. It also cannot be disolved in anything that won't also destroy your tissue samples. I know as I've worked with it for about 20 + years. Now that I've ruined you morning coffee....... try the following.
Remove ALL of the partially polymerized GMA and any water. Place in fresh GMA (no water) at 4*C for 12 hours under vacuum (preferably with agitation). Repeat this step twice more. When ready to embed the tissue allow the polymeization to occur at 4*C. Polymerization should be complete after about 4 hours. Feel free to call me at the numbers listed in my sig line at the end of this message if you have any further questions. -- Begin original message --
} Hi all, } A post-doc has presented me with an interesting task and I would } appreciate some help! I have been given some mouse femurs } (undecalcified) which were placed in a 1:1 mixture of JB-4 resin (both } components A and B in the appropriate mix) and water. The samples were } then placed in a vacuum at 20*C and left over the weekend with the aim } of enhancing infiltration. On Monday the samples were sitting in a } gelatinised resin yuk!! My task is to re-embed them. I am } currently trying to dissolve the resin out with many changes of } distilled water and constant movement but all that has happened is that } the resin has turned white. Any advice would be welcome as these are } "valuable" (aren't they all!) samples.
} Sarah Ellis } } Ps. Alcohol cannot be used. }
-- End original message --
Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chaple Hill, NC 27599 Phone 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
I am desperately looking for a used plunge freezing unit that will take liquid propane. It is for freezing algal cells. I don't want to slam them as they are spherical (even with backing I don't think this would work well....comments?).
So if ANYONE has ANY information on this could you PLEASE let me know!!!
I will be most grateful
Cheers.
Lilian Alessa Postdoctoral Fellow, Kropf Lab Department of Biology University of Utah Salt Lake City, Utah U.S.A.
I need desplastificate slices of 4-5 um of insects legs, which are embedded in Durcupan. I have tryed with methoxide of Sodium, and saturated solution of Ethoxide of sodium ( from 1h. to 24 hrs.), and I couldn't. I have problems with the Ethoxide solution too, it become brown after two days. Somebody knows wath can I do?. Thanks, Veronica. Veronica Campanucci -------------------------------------------------------------------------------- Veronica Andrea Campanucci Laboratorio de Fisiologia de Insectos Dpto. de Ciencias Biologicas Facultad de Ciencias Exactas y Naturales Tel (54 1) 781-5021 to 29, ext. 332 Universidad de Buenos Aires FAX (54 1) 782-0582/544-7893 (1428) Buenos Aires e-mail: veronica-at-bg.fcen.uba.ar Argentina HTTP://biolo.bg.fcen.uba.ar/physinse.htm --------------------------------------------------------------------------------
Do you mean they were put into solution A plus catalyst? In any of the procedures for JB-4 that I have come across, the solution B is only added at the hardening step. The catalyst only encourages the plastic to set up in a quick and orderly manner, but the solutions A and B are the primary components of the plastic. If they are both in the mixture, eventually you can get hardening in any closed container and placing it into a vacuum only hastened the process. I'm not suprized at the reaction with water that you got. Typically, I float fresh-cut sections on water to stretch them and get them to stick to the slide. The JB-4 gets sticky, but doesn't entirely dissolve. I haven't had a block go gooy on me, but it might help if you put it in a fresh change of soltuion A without any solution B or catalyst added. I assume you have manualy removed as much JB-4 from the tissue as you can. Good Luck.
Karen Pawlowski
On Wed, 20 Aug 1997, ellis, sarah wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hi all, } A post-doc has presented me with an interesting task and I would } appreciate some help! I have been given some mouse femurs } (undecalcified) which were placed in a 1:1 mixture of JB-4 resin (both } components A and B in the appropriate mix) and water. The samples were } then placed in a vacuum at 20*C and left over the weekend with the aim } of enhancing infiltration. On Monday the samples were sitting in a } gelatinised resin yuk!! My task is to re-embed them. I am } currently trying to dissolve the resin out with many changes of } distilled water and constant movement but all that has happened is that } the resin has turned white. Any advice would be welcome as these are } "valuable" (aren't they all!) samples. } } Thanks in advance, } } Sarah Ellis } } Ps. Alcohol cannot be used. }
When doing standardless quant. work on the EDX, with an accelerating voltage of 30 KeV, and getting K, L & M lines, what is the rule to follow as far as selecting lines to use in the analysis. Am I supposed to pick the ones with the most counts? Should I pick all of the same, as in Hg-K, Cu-K, Si-K etc? Should I pick the highest energy ones of those detected Because as I'm sure most of you are aware, I get completely different results when I pick the different sets of lines. Mark Darus
You can also keep your EtOH dry by using Drying Beads (Molecular Sieves). We keep ethanol in 200-500 ml bottles with the beads, and bake them out every time we use up the 200-300 ml of ethanol. This saves discarding your expensive abs. ethanol when the bottle has been open for a while. (Caution: make sure you pour out the beads into a pan and let the ethanol evaporate entirely before putting them into the oven, or they will explode and fly EVERYWHERE inside the oven.) Also keep the lid on the bottle except when actually removing some to prevent H2O condensation inside.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Digital Instruments (DI) will be hosting two free workshops on Scanning Probe Microscopy in the New England area. Those interested in attending can find further details at the DI web site www.di.com.
I realize this is only a VERY distantly related question, but I appreciate any help you can give me.
Yesterday I was asked if I know of a source of strepavidin coated microscope slides. One of the investigators I work with wants to use biotinilated cDNA probes and do some confocal microscopy on the result (apparently with other fluorescent probes). His previous attempts have had most of the DNA not stick to the slide through the entire thermocycling process so he's looking for another way to do it. Any ideas on a source for such slides?
TIA, Doug ..................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Sr. Research Specialist 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
Message-Id: {3.0.1.32.19970820101536.0095b100-at-ccit.arizona.edu} X-Sender: dcromey-at-ccit.arizona.edu X-Mailer: Windows Eudora Light Version 3.0.1 (32)
Hi everyone,
The Webmaster of our server tells me that there have been a LARGE number of failed page requests for the Microscopy Pages that I posted last week. I'm sorry for the problems, the long URLs I have to work with apparently wrapped to a second line on many people's email readers & when they tried to access the pages the URL was missing the last few characters (computers are SO literal).
If you're still interested in checking out my series of WWW pages on topics such as general microscopy, histology, confocal, electron microscopy, digital imaging and a list of free publications that are of interest to microscopists you can get to them by this much shorter URL (the links are in the middle of this page): http://www.pharmacy.arizona.edu/exp_path.html
Thanks for your patience.
Yours, Doug Cromey ..................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Sr. Research Specialist 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
I don't know if all manufacturers have this reported in the results, but my TN Voyager gives me data for "Atom %" and "Element %", in many cases the element in majority is different. Can someone explain what these 2 mean, or at least tell me where to go to find out. Thanks
} I don't know if all manufacturers have this reported in the } results, but my TN Voyager gives me data for "Atom %" and "Element %", } in many cases the element in majority is different. Can someone explain } what these 2 mean, or at least tell me where to go to find out. ...
X-ray analysis is primarily sensitive to weight percent ... e.g., lead sulfide (PbS) Pb:S = 87:13 ... however, there is generally always a software option to cast wt% as atomic percent ... i.e., Pb:S = 50:50. Does this answer your question??
cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
Dear Colleagues: A technician here has difficulty to stain mouse CD4 cells by immunofluorecense using an antibody of anti-mouse CD4 conjugated with biotin, and then FITC-avidin. My suspicion is that the problem is with the antibody. As far as I know of, some antibodies are not suitable for immunocytochemistry even if then work well in other tests like ELISA or Immunoblotting. Could some of you recommend some antibody source where we can purchase such Abs against mouse CD4 and CD8 antigen that work well in immunocytochemistry? Thank you very much in advance. Regards, Yuhui Xu, MD,PhD EM Core, DFCI
Lilian, You might be interested in the paper we just published in Microscopy Research & Tech; S. D. Fields, G. W. Strout, & S. D. Russell. Spray-Freezing Freeze Substitution (SFFS) of Cell Suspensions for Improved Preservation of Ultrastructure Micros. Res. & Tech. 38:315-328 1997. The spray freezing device we've developed is decribed within. We have successfully frozen algal and other cells with this method. Greg
Lilian Alessa (by way of Nestor J. Zaluzec) wrote: } } } } Greetings All!! } } I am desperately looking for a used plunge freezing unit that will take } liquid propane. It is for freezing algal cells. I don't want to slam } them as they are spherical (even with backing I don't think this would } work well....comments?). } } So if ANYONE has ANY information on this could you PLEASE let me know!!! }
-- ======================================================== Greg Strout Electron Microscopist, University of Oklahoma e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not neccessarily those of the University of Oklahoma ========================================================
A colleague of mine (no, it was NOT me) mistakenly ran some RC prints through a heated drum dryer with predictable results: the plastic melted onto the drum. Now this person was wondering how to remove the mess. My suggestion: use water soaked towels to remove the paper part and acetone to dissolve the plastic. Any other possibilities? Thanks.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
My EDX system is different, but I would assume that manufacturers pretty much use the same nomenclature and means of obtaining results. It is my understanding that "Element %" is the weight percent calculated for each element, and is not normalized to 100%. This value is a good way to check your analysis; the total % of all elements should not deviate from 100% by very much (I use + or - 2%) unless you have a problem with your analysis. "Atom %" is atomic percent, which is determined by taking the weight percent of each element and dividing by its atomic weight, normalizing, and then determining the atomic percent. This is probably why you have different results for the majority element.
Regards,
Bob ************************* Bob Citron Chiron Vision Claremont, CA USA (909) 399-1311 Bob_Citron-at-cc.chiron.com *************************
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I don't know if all manufacturers have this reported in the results, but my TN Voyager gives me data for "Atom %" and "Element %", in many cases the element in majority is different. Can someone explain what these 2 mean, or at least tell me where to go to find out. Thanks
Does anyone know the approximate amount of protein in solution which would jell a glutaraldehyde-based fixative solution? I have seen this phenomenon occasionally in the past when I was working on animal material where there was alot of cellular damage as part of the experimental design. But, now I've seen this phenomenon while preparing fruit samples (berries) and I don't quite know what to make of it, since glutaraldehyde is a protein crosslinker. We estimate the protein content of our samples to vary between 1 and 10%.
Thanks for any ideas.
Paula.
Paula Allan-Wojtas Food Microstructure Specialist Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
Dear Mark, The general rule is to use a K line when you can, since they are the best characterized and have the best gaussian profile. L's are next and quant. analysis using M lines is usually imprecise. Of course, you must have sufficient overvoltage for the line you want to analyse, try for at least twice the line energy in your acc. voltage (hence a 10 keV x-ray range and a 20 kV acc. voltage). I have not had much luck doing standardless quant. at 30 kV and a 0 to 20 keV range. I find 20 kV and a 0 to 10 keV to be better. Do lots of work with known samples close to the content of your unknowns. BTW, you'll have trouble getting the Hg-K line, it has an energy around 78 keV. You wrote:
} When doing standardless quant. work on the EDX, with an accelerating } voltage of 30 KeV, and getting K, L & M lines, what is the rule to } follow as far as selecting lines to use in the analysis. Am I supposed } to pick the ones with the most counts? Should I pick all of the same, as } in Hg-K, Cu-K, Si-K etc? Should I pick the highest energy ones of those detected Because as I'm sure most of you are aware, I get completely different results } when I pick the different sets of lines. } Mark Darus } } G. E. Lighting Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
} Date: Wed, 20 Aug 1997 22:56:24 -0700 } To: "Mark E. Darus (216) 266-2895 General Electric Co." {darus-at-cle.dnet.ge.com} } From: Mary Mager {mager-at-unixg.ubc.ca} } Subject: Re: Lines for Quant. work } Cc: Microscopy } } Dear Mark, } The general rule is to use a K line when you can, since they are the best characterized and have the best gaussian profile. L's are next and quant. analysis using M lines is usually imprecise. Of course, you must have sufficient overvoltage for the line you want to analyse, try for at least twice the line energy in your acc. voltage (hence a 10 keV x-ray range and a 20 kV acc. voltage). I have not had much luck doing standardless quant. at 30 kV and a 0 to 20 keV range. I find 20 kV and a 0 to 10 keV to be better. Do lots of work with known samples close to the content of your unknowns. BTW, you'll have trouble getting the Hg-K line, it has an energy around 78 keV. } You wrote: } } } When doing standardless quant. work on the EDX, with an accelerating } } voltage of 30 KeV, and getting K, L & M lines, what is the rule to } } follow as far as selecting lines to use in the analysis. Am I supposed } } to pick the ones with the most counts? Should I pick all of the same, as } } in Hg-K, Cu-K, Si-K etc? Should I pick the highest energy ones of those detected Because as I'm sure most of you are aware, I get completely different results } } when I pick the different sets of lines. } } Mark Darus } } } } G. E. Lighting } Regards, } Mary } } Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
Hi We do a lot of diffraction pattern interpretation. Measuring the patters are a tedious job, and in polycrystalline materials it is difficult to measure ring patterns accurately. I have a few questions: *Is there any software available to help this process? That is measuring 'a' as well as the option to try fit the values to standard elemental values from the JCPDS tables. *Does new TEM's (eg Philips CM series) have a measuring option in their software? * If such programs exists, what's the accuracy/resolution of it?
Thanx Sara
-------------------------------------------------------------------------------------- Sara Prins Surface and Structure Analytical Services Division for Materials Science and Technology CSIR PO Box 395 Pretoria South Africa Tel: +27+12+8413974 Fax: +27+12+8414395 sprins-at-csir.co.za Visit us at : http://www.csir.co.za
One problem associated with Molecular Sieve in ethanol is the possibility of dust from the sieve being released into the ethanol. This dust then sticks to the specimen and eventually damages the knife. We have stopped using a drying agent in the ethanol, rather decanting ethanol into smaller (250ml) containers and replacing at regular intervals - saves on diamond knife resharpening!
} } You can also keep your EtOH dry by using Drying Beads (Molecular } Sieves). We keep ethanol in 200-500 ml bottles with the beads, and bake } them out every time we use up the 200-300 ml of ethanol. This saves } discarding your expensive abs. ethanol when the bottle has been open for } a while. (Caution: make sure you pour out the beads into a pan and let } the ethanol evaporate entirely before putting them into the oven, or they } will explode and fly EVERYWHERE inside the oven.) Also keep the lid on } the bottle except when actually removing some to prevent H2O condensation } inside. } } Sara E. Miller, Ph. D. } P. O. Box 3020 } Duke University Medical Center } Durham, NC 27710 } Ph: 919 684-3452 } FAX: 919 684-8735 }
Prof Jan Coetzee Head: Unit for Electron Microscopy Tel:+27-12-420-2075 University of Pretoria Fax:+27-12-342-1738 Pretoria 0002 Internet:janc-at-ccnet.up.ac.za South Africa http://www.up.ac.za/science/electron/emunit1.htm
} One problem associated with Molecular Sieve in ethanol is the } possibility of dust from the sieve being released into the ethanol. } This dust then sticks to the specimen and eventually damages the } knife. We have stopped using a drying agent in the ethanol, rather } decanting ethanol into smaller (250ml) containers and replacing at } regular intervals - saves on diamond knife resharpening!
In order to eliminate the dust problem we place the molecular sieve in a small piece of dialysis tube that is closed using metal clips. We have no water and no dust in acetone, methanol, ethanol. An other way to get rid of water in solvents is to add a small amount of acidified 2,2-Dimethoxypropane. The DMP will react with water to produce ethanol and acetone. If you do not mind small amounts of acetone and DMP in your ethanol this is a very simple way to dry it.
A first guess might be that you've got pectin - of jam and wine haze fame. If the berries make good jam that might be the answer. Malcolm Haswell e.m unit University of Sunderland UK ----------
Hello, everyone,
Does anyone know the approximate amount of protein in solution which would jell a glutaraldehyde-based fixative solution? I have seen this phenomenon occasionally in the past when I was working on animal material where there was alot of cellular damage as part of the experimental design. But, now I've seen this phenomenon while preparing fruit samples (berries) and I don't quite know what to make of it, since glutaraldehyde is a protein crosslinker. We estimate the protein content of our samples to vary between 1 and 10%.
Thanks for any ideas.
Paula.
Paula Allan-Wojtas Food Microstructure Specialist Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
} One problem associated with Molecular Sieve in ethanol is the } possibility of dust from the sieve being released into the ethanol. } This dust then sticks to the specimen and eventually damages the } knife. We have stopped using a drying agent in the ethanol, rather } decanting ethanol into smaller (250ml) containers and replacing at } regular intervals - saves on diamond knife resharpening!
This discussion has been brought up previously on the list. Many labs encase the molecular sieves in dialysis tubing to prevent the dust from contaminating the solution.
Dennis Shubitowski University of Michigan dshubito-at-umich.edu
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Memo : job opp.: intermetallics 21-08-1997 15:48
Hi all,
We're looking for a TEM specialist with practical knowledge of intermetallics for a 6 or 12 month (Jan. - Dec. 1998) post-doc position at the Electron Microscopy for Materials Science group in Antwerp, Belgium, and working on Ni- and Fe-based materials.
If you're interested please contact me as soon as possible. The application can only be started when the applicant is known.
Thanks,
Nick Schryvers
Dr. Dominique Schryvers University of Antwerp, RUCA - EMAT Groenenborgerlaan 171, B-2020 Antwerpen (Belgium) Tel: 32-3-2180247 Fax: 32-3-2180257 e-mail: schryver-at-ruca.ua.ac.be homepage: http://www.ruca.ua.ac.be/~EMAT/Schryvers.html
The rule of thumb to follow if possible use the lines in order K, then L then M. If using standardless analysis you want to use the lines in the same family if possible preferably all K lines. Be careful of overvoltage if using a 30kV beam for some of the lower energy lines. Is 30kV necessary to excite all of the lines that you want? The rule of thumb is to use at least twice the energy of the line as your accelerating voltage.
My experience with standardless analysis has also been that there is less acuracy with increaseing elements. Brasses and Bronzes come up very well but the accuracy trails off as you add more elements. And super alloys are almost impossible. I hope that this has been hepful.
As to the question of weight percent and atom percent. The atom percent is basicaly what percentage of atoms of one element there are in relation to another (ie a 1:1 ratio of Cu to Zn would produce a 50% at% Cu and 50 at% Zn where a 7:3 ratio would produce a 70 at% Cu to 30 at% Zn). The weight percent takes into account the atomic number of the elements or their atomic weight therfore a 50 at% Al and 50 at% W would show a weight percent much higher for the W atom since it is much heavier. I hope this has been helpful. Please let me know if you have anymore questions.
__________________________ Roberto Garcia EMF Manager Wright State University rgarcia-at-cs.wright.edu
I have to say that when I have seen light/dark bands on slow scan (mainly textiles) I usually find that the problem is charging on the sample--not a microscope problem. I believe it to be a capacitance effect where the sample charges and dissipates. Usually, recoating the sample with gold or using Fullam's antistatic liquid (I hang my head in shame!) solves the problem.
Changing the voltage may help troubleshoot the problem as well.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Olli,
They kept trying to say that my problems were fields, but the active field cancellation could not affect the visual symptoms. The ultimate proof that it was inside the system, is the fact that the problems went away when they replaced the entire column. It is easy for SEM manufacturers to blame fields for problems they can not trace, or easily solve. They all seem to use this crutch from time to time.
Do not get me wrong, I know fields can be a problem. My response to that is that, as the systems become more and more susceptible to the ocean of magnetic noise they live in, the manufacturers need to prevent the affects with their design, and not use it as an excuse for poor performance in the real world.
Enough of that! I was just wondering if anyone else was having these problems on similar systems. It seems as though there are a few out there.
I wanted to find out if anyone could tell me if they use LR White as an embedding resin NOT for immuno work. The reason why I say not for immuno work, is that I would like to osmicate them. I was curious to see if I fix samples in Gluteraldehyde, and then osmicate them, could I embed them up in LR white with Gelatin capsules/coverslips and then section and stain for the TEM. How does this resin hold up under the beam? I haven't seen any one mention this on this listserver, and wondered if people even do this. Generally, I use Spurrs resin, but have some extra LR White resin that I would like to use up before it expires.
Susan Carbyn Atlantic Food and Horticulture Research Centre Agriculture and Agri-Food Canada Kentville, Nova Scotia B4N 1J5 Canada
Just wanted to report that using a fresh bottle of absolute ETOH has solved the "pinhole" artifact ... such an easy fix for such an irritating problem. Thanks again for your help. (The additional conversation about the sieves has also been enlightening.) Sharron G. Chism HT (ASCP) Electron Microscopy Lab Harris Methodist Hospital Fort Worth, Texas
In message {s3fc16e2.081-at-EM.AGR.CA} Susan Carbyn writes:
} I wanted to find out if anyone could tell me if they use LR White as an } embedding resin NOT for immuno work. The reason why I say not for } immuno work, is that I would like to osmicate them. I was curious to see } if I fix samples in Gluteraldehyde, and then osmicate them, could I embed } them up in LR white with Gelatin capsules/coverslips and then section } and stain for the TEM.[?]
Sure, and because of its very low viscosity, LR White works well for infiltrating into plant tissues.
} How does this resin hold up under the beam?
Not as well as epoxy sections do, on uncoated grids. They can drift, tear or flap in the electron "breeze". Solution: 1. use Formvar or similarly coated grids to stabilize the sections. 2. Coat LR White sections (mounted on bare grids) with thin carbon layer in a vacuum evaporator, taking care to minimize heat delivered to sections. 3. Use higher mesh grids, eg. 200-400#.
} Susan Carbyn } Atlantic Food and Horticulture Research Centre } Agriculture and Agri-Food Canada } Kentville, Nova Scotia B4N 1J5 } Canada } } E-mail: carbyns-at-em.agr.ca } } Phone: (902) 679-5566 } Fax: (902) 679-2311
Good luck!
Gib
--
Gib Ahlstrand, MMS Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
LR White is a good embedding medium for staright morphology after osmication. Doesn't always section as nicely as an epoxy, but still very useful for hard to embed materials. } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } At 10:18 AM 8/21/97 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America Scientific Director, ICBR Electron Microscopy Core Lab PO Box 118525 Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
Susan Carbyn wrote: } } Just a quick question: } } I wanted to find out if anyone could tell me if they use LR White as an } embedding resin NOT for immuno work. The reason why I say not for } immuno work, is that I would like to osmicate them. I was curious to see } if I fix samples in Gluteraldehyde, and then osmicate them, could I embed } them up in LR white with Gelatin capsules/coverslips and then section } and stain for the TEM. How does this resin hold up under the beam? I } haven't seen any one mention this on this listserver, and wondered if } people even do this. Generally, I use Spurrs resin, but have some extra } LR White resin that I would like to use up before it expires. } } Susan, You can use the LR White as long as you cure the resin in the oven. The Osmium will not cause any problems. The resin holds up fine in the beam. Just remember to exclude exposure to oxygen or the resin will not cure.
Greg Rudomen Greg-at-umic.sunysb.edu University Microscopy Imaging Center SUNY Stony Brook
I had some misfortunes with osmium-fixed tissue embedded in LR White resin. The resin polymerized prematurely during infiltration, but it worked well with glut-fixed tissues. Someone thought that I had too much an accelerator. The real problem was that LR White was too old and it reacted with osmium. A new bottle of resin worked well for a while and then it acted up again.
I think the way to counter this problem is to buy LR White without the accelerator already mixed. When needed, one can mix them up and divide into several portions for storage. A portion will be warmed up each time and it can be used up quickly. The rest of the resin stays cold, therefore, it can be kept for a long time without causing problems.
Ann Fook Yang, EM Unit, ECORC, Agriculture and Agri-Food Canada, Ottawa, Ontario, Canada K1A 0C6
} } } Susan Carbyn {CarbynS-at-em.agr.ca} 08/21/97 10:18am } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Just a quick question:
I wanted to find out if anyone could tell me if they use LR White as an embedding resin NOT for immuno work. The reason why I say not for immuno work, is that I would like to osmicate them. I was curious to see if I fix samples in Gluteraldehyde, and then osmicate them, could I embed them up in LR white with Gelatin capsules/coverslips and then section and stain for the TEM. How does this resin hold up under the beam? I haven't seen any one mention this on this listserver, and wondered if people even do this. Generally, I use Spurrs resin, but have some extra LR White resin that I would like to use up before it expires.
Susan Carbyn Atlantic Food and Horticulture Research Centre Agriculture and Agri-Food Canada Kentville, Nova Scotia B4N 1J5 Canada
We use LR White for non-immuno routinely. We use grids without = supporting membranes for both LR White and Embed. It holds up in the = beam, although is a bit less stable than Embed or any of the other Epon = replacements and is only a problem with some of the newest students who = let a crossover beam sit on it. Gelatin capsules can be used and some people also use BEEM capsules. = Some of the BEEM type capsules however are more permiable to oxygen, but = others seem to work OK. It is important that all the EtOH be removed = before embedding. As such we do not use EtOH:resin, 2:1, 1:1, and 1:2 as = we do in Embed. We go through 100% EtOH twice, then into 3 changes of = pure LR White. We love it and all the students love it. I personally have used LR White = since before it was actually an EM product because it is so fast and easy = to use. Hope comments are helpful. Judy M.
Judy Murphy, PhD Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207
Phone: 209/954-5284 FAX: 209/954-5600 e-mail: jmurphy-at-sjdccd.cc.ca.us program web page: http://www.sjdccd.cc.ca.us/ElectMicro/sjdc.html
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Just a quick question:
I wanted to find out if anyone could tell me if they use LR White as an embedding resin NOT for immuno work. The reason why I say not for immuno work, is that I would like to osmicate them. I was curious to see if I fix samples in Gluteraldehyde, and then osmicate them, could I embed them up in LR white with Gelatin capsules/coverslips and then section and stain for the TEM. How does this resin hold up under the beam? I haven't seen any one mention this on this listserver, and wondered if people even do this. Generally, I use Spurrs resin, but have some extra LR White resin that I would like to use up before it expires.
Susan Carbyn Atlantic Food and Horticulture Research Centre Agriculture and Agri-Food Canada Kentville, Nova Scotia B4N 1J5 Canada
E-mail: carbyns-at-em.agr.ca
Phone: (902) 679-5566 Fax: (902) 679-2311
------------------ RFC822 Header Follows ------------------ Received: by sjdccd.cc.ca.us with ADMIN;21 Aug 1997 10:28:24 -0800 Received: from Sparc5.Microscopy.Com (206.69.208.10) by = ms.sjdccd.cc.ca.us with SMTP (Apple Internet Mail Server 1.1.1); Thu, 21 Aug 1997 10:28:12 = -0800 Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com = (8.6.11/8.6.11) id JAA02109 for dist-Microscopy; Thu, 21 Aug 1997 = 09:20:01 -0500 Received: from agrout1.agr.ca (agrout1.agr.ca [192.197.71.131]) by = Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id JAA02106 for = {Microscopy-at-sparc5.microscopy.com} ; Thu, 21 Aug 1997 09:19:59 -0500 Received: from agrgate.agr.ca (agrgate.agr.ca [192.197.71.189]) by agrout1.agr.ca (8.8.4/8.8.4) with SMTP id KAA23603 for {Microscopy-at-sparc5.microscopy.com} ; Thu, 21 Aug 1997 = 10:22:58 -0400 (EDT) Received: from agrin1.agr.ca by agrgate.agr.ca via smtpd (for agrout1.agr.ca [192.197.71.131]) with SMTP; 21 = Aug 1997 14:31:23 UT Received: from EM.AGR.CA (gwiseweb.agr.ca [142.61.33.38]) by agrin1.agr.ca (8.8.4/8.8.4) with SMTP id KAA24906 for {Microscopy-at-sparc5.microscopy.com} ; Thu, 21 Aug 1997 = 10:24:37 -0400 (EDT) Received: from AGCAN-Message_Server by EM.AGR.CA with Novell_GroupWise; Thu, 21 Aug 1997 10:22:26 -0400 Message-Id: {s3fc16e2.081-at-EM.AGR.CA} X-Mailer: Novell GroupWise 4.1
Hi,
LR White is an acrylic embedding medium. It has many advantages for immuno work, among them its low crosslinkage. It does not bind with the tissue (like epoxies) but through tissue. It does not preserve tissue as well as epoxy. It is not as beam stable as epoxy. Its polymerization reaction is exothermic - if uncontrolled - it may damage tissue. Thick sections may wrinkle badly (due to the lack of crosslinkage). Simply to use LR White because it is in the refrigerator is not a good idea. For non-immuno work it is far more advatageous to use epoxy monomers. Bye, Hildy
A quick note of congratulations on the success of the 2-Photon seminar at last week's Microscopy & Microanalysis meeting. I only had a chance to visit for the last afternoon, but it was very clear that the sessions were well attended and that the vendors got a lot of opportunity to show off their new gear and run samples.
Is there a way that those of us who could not attend the regular workshop could get a copy of the notes? Please post info on both listservers so that a wider audience can respond.
Thanks! Barbara Foster Microscopy/Microscopy Education
I am in search of the Representative on Reichert-Jung Miccrotomes in the Seattle area. We are interested in looking at a reichert ultracut microtome?? anyone with the address or phone number wold be appreciated
Eric A.Rosen Fred Hutchinson Cancer Research Center
This is a request for information from you LM types out there. Does anyone have any suggestions for the conversion from polaroid to digital outlined below? I'm sure this has been discussed but I probably ignored it since I don't deal with LM much, but I told these people I'd send this message out for them. The $15k has to cover the computer as well. Bruce said he has information on a Polaroid system and a Leco setup. Any others? We don't have a great preference between Mac and Wintel systems as we run both.
I'd appreciate any responses. Thanks in advance for the help.
Cheers,
John Vetrano Materials Interfaces Group Pacific Northwest National Laboratory _______________________________________________________________________________
John, we have $15k to purchase CCD camera for the Zeiss and Olympus microscope in metallography. We are looking to replace the polaroid camera system in place with a CCD camera. Both scopes have C-mounting capability. Will need a CCD camera for high quality metallograpy work. We would like to capture the image and store the image for future retrieval, we would like a similar system like the SEM (Gatan DigiScan). Thanks for offering to place a ad on the server list.
On Thu, 21 Aug 1997, Jan Coetzee EM Univ Pretoria wrote:
} Date: Thu, 21 Aug 1997 10:50:16 CAT-2 } From: Jan Coetzee EM Univ Pretoria {janc-at-ccnet.up.ac.za} } To: Microscopy-at-sparc5.microscopy.com } Subject: RE: TEM Artifact } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } One problem associated with Molecular Sieve in ethanol is the } possibility of dust from the sieve being released into the ethanol. } This dust then sticks to the specimen and eventually damages the } knife. We have stopped using a drying agent in the ethanol, rather } decanting ethanol into smaller (250ml) containers and replacing at } regular intervals - saves on diamond knife resharpening! } JUST LET THE BOTTLE SIT, AND THE DUST WILL SETTLE TO THE BOTTOM. DON'T DISTURB IT WHEN REMOVING SOME AND TAKE IT OFF THE UPPER LAYER. WE NEVER HAVE ANY PROBLEMS WITH DIRTY SAMPLES OR SHORTENED DIAMOND KNIFE LIFE.
AS WITH ANY REAGENT, IF IT LOOKS CLOUDY, DISCOLORED, OR UNUSUAL, I DON'T USE IT.
S. MILLER
} } } } You can also keep your EtOH dry by using Drying Beads (Molecular } } Sieves). We keep ethanol in 200-500 ml bottles with the beads, and bake } } them out every time we use up the 200-300 ml of ethanol. This saves } } discarding your expensive abs. ethanol when the bottle has been open for } } a while. (Caution: make sure you pour out the beads into a pan and let } } the ethanol evaporate entirely before putting them into the oven, or they } } will explode and fly EVERYWHERE inside the oven.) Also keep the lid on } } the bottle except when actually removing some to prevent H2O condensation } } inside. } } } } Sara E. Miller, Ph. D. } } P. O. Box 3020 } } Duke University Medical Center } } Durham, NC 27710 } } Ph: 919 684-3452 } } FAX: 919 684-8735 } } } } } } Prof Jan Coetzee } Head: Unit for Electron Microscopy Tel:+27-12-420-2075 } University of Pretoria Fax:+27-12-342-1738 } Pretoria 0002 Internet:janc-at-ccnet.up.ac.za } South Africa http://www.up.ac.za/science/electron/emunit1.htm }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
} --Paula, Not 5 minutes before your e-mail I was asked to look at some artery from a goat which was showing exactly what you described. The vessel had been incubated with Clostridium perfrinogen toxin,it was covered with a gelatenous substance. But I cant help you answer your question.
Christine Lee, Vet. Pathobiology, University of Queensland. -
--------------------------------------------------------------------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Eric, Reichert -Jung is now called Leica their # is 800-248-0123. -- Begin original message --
} From: Eric Rosen {erosen-at-fred.fhcrc.org} } Date: Thu, 21 Aug 1997 13:33:45 -0700 (PDT) } Subject: Microtome manufacturer address } 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 } -----------------------------------------------------------------------. } } } I am in search of the Representative on Reichert-Jung Miccrotomes in the } Seattle area. We are interested in looking at a reichert ultracut } microtome?? anyone with the address or phone number wold be appreciated } } Eric A.Rosen } Fred Hutchinson Cancer Research Center } } } }
-- End original message --
Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chaple Hill, NC 27599 Phone 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
Ann-Fook Yang (Ann-Fook Yang) wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } I had some misfortunes with osmium-fixed tissue } embedded in LR White resin. The resin } polymerized prematurely during infiltration, but it } worked well with glut-fixed tissues. Someone } thought that I had too much an accelerator. The } real problem was that LR White was too old and it } reacted with osmium. A new bottle of resin worked } well for a while and then it acted up again. } } I think the way to counter this problem is to buy LR } White without the accelerator already mixed. } When needed, one can mix them up and divide into } several portions for storage. A portion will be } warmed up each time and it can be used up } quickly. The rest of the resin stays cold, therefore, } it can be kept for a long time without causing } problems. } } Ann Fook Yang, } EM Unit, } ECORC, Agriculture and Agri-Food Canada, } Ottawa, Ontario, Canada } K1A 0C6 } } } } } Susan Carbyn {CarbynS-at-em.agr.ca} } 08/21/97 10:18am } } } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Just a quick question: } } I wanted to find out if anyone could tell me if they } use LR White as an } embedding resin NOT for immuno work. The } reason why I say not for } immuno work, is that I would like to osmicate them. } I was curious to see } if I fix samples in Gluteraldehyde, and then } osmicate them, could I embed } them up in LR white with Gelatin } capsules/coverslips and then section } and stain for the TEM. How does this resin hold up } under the beam? I } haven't seen any one mention this on this listserver, } and wondered if } people even do this. Generally, I use Spurrs resin, } but have some extra } LR White resin that I would like to use up before it } expires. } } Susan Carbyn } Atlantic Food and Horticulture Research Centre } Agriculture and Agri-Food Canada } Kentville, Nova Scotia B4N 1J5 } Canada } } E-mail: carbyns-at-em.agr.ca } } Phone: (902) 679-5566 } Fax: (902) 679-2311
Dear Ann-Fook Yang, We agree with you. Ladd sells LR White and about two years ago we stopped selling it with the accelerator already mixed in for the reason you stated. John Arnott
In response to a question on glutaraldehyde, the name of the Ladd Research's head chemist, Dr. Charles Duvic was indvertently entered as "Garber". Ladd apologizes profusely to those who contacted us concerning this error. Dr. Duvic has worked for many years to develop the quality and reputation of Ladd's glutaraldehyde and other chemicals, so it is no way an insult to have ones name substituted for his. Never the less we do apologize to any one who was offended.
Information regarding the International EM Congress next year may be found at:
http://icem.inin.mx
Take a look ** Alwyn Eades Center for Microanalysis of Materials University of Illinois at Urbana-Champaign Phone 217 333 8396 Fax 217 244 2278 eades-at-uimrl7.mrl.uiuc.edu (NB those are letter l not ones) **
I, too, had an incident where my osmium-fixed tissue caused the LR White to polymerize during an infiltration step. I was also infiltrating samples of the same tissue without osmium and had no early polymerization problems. We have not had any other problems with LR White aside from the occasional oxygen inhibiting polymerization a bit, but we don't usually osmicate samples for LR White embedding. I did not add accelerator at any point during the infiltration and all of the samples were at the same temperature. I have not had the time to follow-up on the issue, yet.
Gregg Sobocinski Parke-Davis Pharmaceutical Research Division Ann Arbor, Michigan, USA Sobocig-at-aa.wl.com -------------------------------------------------------------------. } } I had some misfortunes with osmium-fixed tissue } embedded in LR White resin. The resin } polymerized prematurely during infiltration, but it } worked well with glut-fixed tissues. Someone } thought that I had too much an accelerator. The } real problem was that LR White was too old and it } reacted with osmium. A new bottle of resin worked } well for a while and then it acted up again. } } I think the way to counter this problem is to buy LR } White without the accelerator already mixed. } When needed, one can mix them up and divide into } several portions for storage. A portion will be } warmed up each time and it can be used up } quickly. The rest of the resin stays cold, therefore, } it can be kept for a long time without causing } problems. } } Ann Fook Yang, } EM Unit, } ECORC, Agriculture and Agri-Food Canada, } Ottawa, Ontario, Canada } K1A 0C6
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} ...Gelatin capsules can be used and some people also use BEEM capsules. } Some } of the BEEM type capsules however are more permiable to oxygen, but } others seem } to work OK...
Flat molds for specimen orientation can be a problem, since most are permeable to LR White. Pella sells a teflon flat mold that solves the problem.
} From Susan Carbyn:
} I wanted to find out if anyone could tell me if they use LR White as an } embedding resin NOT for immuno work.
LR White Hard grade is great for hard biological (bone, keratinized epithelium, etc.) and non-biological (catalysts, hard polymers, etc.) samples.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
Hi all, I'm looking for the power supply for a Zeiss Photomicroscope III (Zeiss p/n 47 20 83). If you have one for sale, trade, or donation, please contact me at the address below. TIA Julian
Julian P.S. Smith III Biology Winthrop University Rock Hill, SC 29733 803-323-2111 x227 (vox) 803-323-2246 (fax)
Does anyone know of any sources for antibodies against phosphorylated forms of MAP-2 (microtubule associated protein-2)?
Thanks in advance, Glen MacDonald Virginia Bloedel Hearing Research Center Box 35-7923 University of Washington Seattle, WA 98195-7923 (206) 616-4156 glenmac-at-u.washington.edu *---------------------------------------------------------------------* The box said "Requires Windows 95 or better.", so I bought a Macintosh. *---------------------------------------------------------------------*
Thanks to the ten or so people who replied to my question about bakeout frequency and flash intensity on Hitachi S4500 FESEMs, either directly or via the server. The word ' from the horse's mouth' was that the flash current should be around 25-30uA for a clean tip - higher may indeed damage the tip, and bakeout should be done when the vacuum deteriorates, inter-flash interval decreases too much, or tip noise increases and cant be decreased by running at 30kV for 2-4 hours then 5kV. The range of replies was awesome - one person has been flashing in the forties for years, bakeout recommendations varied from monthly to "not done in 60 months" - but everyone seemed happy with the performance under the regime they were using.
So, we baked out, adjusted the flash intensity (call your service man), and made good resolutions about keeping records of flash current and post flash extraction voltage. And always using the cold trap.
For the record, we are a service unit with a wide range of users, we have a diffusion-pumped chamber and run a cold stage from time to time. } } Hi all, } I am after some advice on the care of the FE tip and vacuum } on a Hitachi 4500. } } 1. How often should one bake out? The manual recommends baking out } when the vacuum deteriorates. After about 8 months operation ours is } better than it was to start with - IP1&2 off-scale, IP3 at } 7x10-7 Pa. On the other hand many people seem to recommend baking at } fairly short intervals "whether it needs it or not". We are inclined } to a non-interventionist approach but are getting a bit nervous...any } advice? } } 2. What should the flash current intensity be? Ours started at around } 15-20 (and we sometimes flashed twice to get a reading in the high } twenties) but has steadily crept up and is now in the high forties. } Is this good, bad or indifferent? Is it perhaps related to question } 1? If it gets too high does it wreck the tip? We are generally } flashing once or twice a day. ---------------------------------------------------------------------- Sally Stowe |Email: stowe-at-rsbs.anu.edu.au Facility Coordinator |Post: ANU Electron Microscopy Unit |ANUEMU (RSBS) Ph 61 6 249 2743 |Australian National Univ. FAX 61 6 249 4891 |Canberra, http://online.anu.edu.au/EMU/home.htm |AUSTRALIA 0200
If you are (*were!*) a member of Histonet, please note the server was disabled by a lightning strike and power outage. The power problems also took out the backup system!
Herb Hagler informs us that things are now back to normal, but former members will have to resubscribe to the listserver.
Send a message to: Histonet-at-pathology.swmed.edu
with the following as the subject: subscribe
Please pass this information to your friends and colleagues who were subscribers to Histonet.
Thank you for informing me about your product but i have a free copy of Evex's Analytical copy of VIDX Microanalysis software. The fellows there have already confiqured my first system for me and they are working to get the second system up and running.
They were able to use my old Kevex pulse processor and power supply and interface directly to a PC using windows 95 or NT. It want even that expensive. I was able to get a demoe unit for less than $10,000.00.
Cheers Craig Ross
purchased the VIDX softwaare Buying software to work with my old equipment does not chnage the fact that the old equipment can still break and that in the long run I might be better off just buying an Evex System. It
. Fortunatelky there are independent organization such as Evex Analytical that service old equipment like my Kevex and other equipment such as Tracors and PGT's that help keep it alive , But Is the software your selling
Personally, I dont believe anything from a person or company that does not sign their name.
My Two Cents Bill Zender
--------------------- Forwarded message:
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Craig Ross wrote: "I am search for old Kevex parts for Kevex 7000 system. Our has bit the dust and we dont have too much money to spend on a new system." Dear Craig, It is very difficult to maintain old EDX systems and new ones are often prohibitively expensive. If your pulse processor is still working, consider upgrading your system to Microsoft Windows as offered by our company and several others. For a free demo software check out our website. Mektech Inc. www.visionol.net/~mektech
Bill Zender wrote: } Personally, I dont believe anything from a person or company that does not } sign their name.
} My Two Cents } Bill Zender
Dear Bill,
It's our company policy to sign with the company's name when mentioning our products on the listserver. This leaves no doubt as to our interests and is far more honest than posing as "EDXUSER"
Today there are many alternatives for the user who has a good detector but an ageing set of electronics.
Just keep the detector: If the detector is still good, one can keep it and replace the bias supply, pulse processor and MCA. A complete replacement (not including a PC of the user's choice) system including quantitative analysis software is available from ANS for $13,990. Replacing the entire electronics package can often improve both the resolution and count rate performance of a system.
Keep the detector, bias supply and pulse processor: Upgrade consists of a new MCA and Windows software. In this case ANS's upgrades can run from $4400 for a semi-Q package to $8,990 for a fully quantitative system (PC supplied by user).
Various companies have different approaches to the upgrade issue. We would recommend that anyone who is considering an upgrade should check out all of the possibilities and download evaluation software (ANS's is available at www.ansxray.com) whenever possible.
Bill Hardy President American Nuclear Systems, Inc. Manufacturer of EDS upgrade packages
Dear Listserver Readers, This is a commercial posting by Thomson Scientific Instruments Pty. Ltd. Whilst I absolutely abhor the practice of commercial postings, the recent thread concerning EDX upgrades is an obvious setup and commercially I am left with no option but to respond.
Given that our competitors have been advertising their wares I would like to point out that we also produce a upgrade package for old EDS systems and have done since 1993.
With sincere apologies to any readers who consider this posting inappropriate,
Paul Thomson Technical Director Thomson Scientific Instruments Australia Web Page: http://werple.net.au/~tsi/
This is written in response to Paul Thomson's posting. While his claim that the recent discussions have been a jackup may be true (if so it went over my head), I have welcomed the contributions from the vendors, including Paul's, as knowledge of what's currently available is always welcome in my book, as long as it's not rammed down my throat. In fact, I would like to invite vendors of upgrade equipment to contact me, as my beloved Link QX2000 is, I'm afraid, unlikely to make it into the millenium, and I'm unlikely to be able to afford a new Oxford system. I want full quantitative analysis and quantitative mapping (preferably real-time), as it will be for an EDS-only EPMA (now there's what some may regard as an oxymoron). Maybe you'd better contact me directly.
thanks
Ritchie
ps does anyone know if anybody has set up a listserver for XRF yet?
cheers
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
There has been a rash of postings the last few days about X-ray Analysis Systems. I appreciate the fact that some of our subscribers are manufacturers and whole heartily support their participation on the Listserver...
However, as you will all recall, one of our cardinal rules is no advertising. This last round of postings started by one vendor and then followed on by a number of others has begun to cross that line. Please, keep you postings to answering questions.
For example, recommendations on keeping a detector and just replacing the electronics are fine. Or stating that you manufacture a product which solves this problem also okay but then direct the reader to your WWW site for details.
However, I do not wish to see items posting that say
......... and for $xxx.yy we can sell you a product that does .........
that type of posting is crossing the line, which I admit is gray, but nevertheless is against the philosophy of this listserver. That is clearly selling rather than giving information.
There have been a number of manufacturers that have done this recently, and a number that have complained to me privately. As many of you know I usually only send out private messages to the company that crosses the line, however in this case I see the potential for many to say , well , I'll do it too. Please refrain from this type of posting.
Thanks
Nestor Your Friendly Neighborhood SysOp/Policeman?
Exerpt from the RULES of the LISTSERVER......
---------------------------------------------------------------------------- Can I post an Advertisement? ----------------------------------------------------------------------------
No, that does not fit within the bounds of this discussion forum.
This listserver is not intended to be a Sales mechanism for commerical organizations, but rather it is an open discussion area about microscopy and microanalysis problems and solutions. If you are an organization and have equipment you wish to donate, or sell, for nominal cost (i.e. no profit) then this is generally an acceptable posting. If you are not sure then send a copy of the announcement in question to Zaluzec-at-MSA.Microscopy.Com and I will give you my opinion. An example of this type would be an old decommissioned instrument which someone is trying to give away for removal/shipping costs, that would fit within the bounds of the purposes of this list.
If you are a manufacturer, you are always welcome to observe/join in any discussion at any times. We do ask that everyone, please refrain from overt sales pitches and/or commericalism. If a product which you produce/sell can solve a problem or answer a question raised by anyone on this list, then by all means feel free to say so. Try to be brief about the product, state the simple facts in a few (short) sentences and then offer to continue the discussion with any interested parties offline. Alternatively you can give sufficient information so that individuals can download/access the relevant information.Usually it will be sufficient to just add your phone number and/or Email address to the end of your message, and you'll be contacted by anyone that is interested.
Remember, please keep your comments about any product you "sell" to a minimum.
It is not out of line to provide your company name, Email address or WWW site as part of your signoff/signature line, at the end of ANY message you post to this system.
This Listserver operates on the honor system with respect to to posting of advertising, so please respect these simple ground rules.
If you are interested in using the Internet for Commerical Advertising of Microscopy/Microanalysis Related Products/Services, you may wish to contact MSA at it's WWW site (http://www.msa.microscopy.com) or the MSA Business Office (MSABusinessOffice-at-MSA.Microscopy.Com). These alternative Internet services, are provided independently of the Listserver Operation, which MSA provides as a FREE service to the WorldWide Microscopy and Microanalysis Community. Any funds derived from the above are used to defray the costs of running MSA's Internet site.
--IMA.Boundary.511815278 Content-Type: text/plain; charset=US-ASCII Content-Transfer-Encoding: 7bit Content-Description: cc:Mail note part
John,
Are you referring to a ferrotyping drum drier? If so, GOOD LUCK at not scratching the surface. I would think that a soft cotton towel (not paper) soaked in a solution of water and wetting agent to remove the paper residue but also make sure that the paper doesn't rub the surface as it comes off. I really think that you better call the paper manufacturer and get their suggestions on removing the coating material.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
A colleague of mine (no, it was NOT me) mistakenly ran some RC prints through a heated drum dryer with predictable results: the plastic melted onto the drum. Now this person was wondering how to remove the mess. My suggestion: use water soaked towels to remove the paper part and acetone to dissolve the plastic. Any other possibilities? Thanks.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 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 ####################################################################
(IMA Internet Exchange 2.1 Enterprise) id 0018E94F; Fri, 22 Aug 97 21:01:57 -0500 Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by ns2.baxter.com (8.8.0/8.8.0) with SMTP id VAA04630; Fri, 22 Aug 1997 21:09:33 -0500 (CDT) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id QAA16145 for dist-Microscopy; Wed, 20 Aug 1997 16:38:58 -0500 Received: from saluki-mail.siu.edu (saluki-mail.siu.edu [131.230.252.17]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id QAA16142 for {microscopy-at-sparc5.microscopy.com} ; Wed, 20 Aug 1997 16:38:57 -0500 Received: from [131.230.97.71] (mac71.chem.siu.edu [131.230.97.71]) by saluki-mail.siu.edu (AIX4.2/UCB 8.7/8.7) with SMTP id QAA10624 for {microscopy-at-msa.microscopy.com} ; Wed, 20 Aug 1997 16:45:52 -0500 (CDT) X-Sender: bozzola-at-saluki-mail.fiber2.siu.edu Message-Id: {v02130502b021154f05c1-at-[131.230.97.71]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
I would appreciate comments from anyone using PhotoShop on a Unix based system. ******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab PO Box 118525 Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
Hi, Can anyone help me for the following problem : I need to visualize waxes on plant cuticles in TEM Are there methods to contrast waxes by chemicals like OSO4 for lipids for instance. I looked for some informations in literature but didn't find something interesting... Lack of time... I need to know about it so quickly as possible Any informations are welcome Thanks to All Pascal
************************************ Pascal VEYS Laboratory of Plant Biology Catholic University of Louvain Place Croix du Sud 5 (bte 14) B 1348 Louvain-la-Neuve Belgium Phone : 0032 10473004 Fax : 0032 10473471 Email : Veys-at-bota.ucl.ac.be ************************************
I believe that there has been mention of an atomic force microscopy listserver at some time on this list. I am at home, recovering from surgery and can't look through my filed mail at the office.
We have begun doing some AFM work and I would like the address of such a server, if it does exist. I'd like to make some good use of my convalescence and learn more about the AFM before I return for work.
Bob Lawrence
"The valley of the spirit never dies; It is the woman, primal mother. Her gateway is the root of heaven and earth. It is like a veil barely seen. Use it; it will never fail."
---------------------------- Forwarded with Changes ---------------------------
Position : Analytical Equipment Engineer ---------------------------------------- Requirement : ------------ - Proficient in both practical and theoretical aspects of Secondary, Backscatter and EDS for Field Emission Scanning Electron Microscopy. - Experience in various Electron Optic equipments (FESEM, Auger and TEM) - Knowledgable in ion beam type equipment (Focus Ion Beam) - Familiar with semiconductor fabrication process and material science. - Bachelor/Master/Phd Degree in Material Science or Electrical Engineering - Good technical skill in beam based FA equipment (SEM / EDX, FIB, or TEM), material science or semiconductor fabrication process
Job Function ------------ - Be a part of Intel's dynamic and technical team performing failure analysis on Intel Microprocessors - Improve product yield, quality and reliability through in-depth failure analysis to identify defect using state of the art FA equipments (SEM/EDX, FIB) and through detailed understanding of fabrication technology - Opportunity to work very closely on Intel advanced multilayered fabrication processes (0.40 um and 0.25 um process technology) - Involved in supporting new product transfer and startup, automate and improve the failure analysis process and proliferate shared learning across Intel sites. - Job requires the condidate to be PERMANTLY STATIONED in INTEL PENANG, MALAYSIA
Candidates please contact:
In Malaysia:
Eng Hong Yeoh Tel: 604-859-6160 FAX: 604-859-6749 e-mail: Eng_Hong_Yeoh-at-ccm.ipn.intel.com
or in the US:
Kian Sin Sim, John Mardinly or David Susnitzky Intel SC2-24 2200 Mission College Blvd. Santa Clara, CA 95052-8119
David Henriks TEL: 800-728-2233 (toll-free in USA) South Bay Technology, Inc. 714-492-2600 1120 Via Callejon FAX: 714-492-1499 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.
Welcome to the Scanning Probe Microscope List/Digest!
This list is intended to be a forum for discussing views, issues, and applications of Scanning Probe Microscopy with the goal of expanding knowledge of SPM and bringing the SPM community closer together. =
You may submit articles to spm-at-di.com.
This is an open forum for exchange of information with a single caveat: =
Posting of proprietary information on any manufacturer's product is not permitted. Offenders will be deleted from the list. Other than that, th= e usual rules of Internet conduct are expected.
If you want to subscribe/unsubscribe to the list/digest, send lines similar to these below to majordomo-at-di.com. Do not include the ' {' or '} ' characters in the message.
(The "end" command prevents majordomo from barfing on your signature.)
Note that you do not need to type in the email address in the above examples if your mailer already puts in a good From: line or Reply-To: line in your mail header. If you don't know what that means, supply your email address.
When it comes time to unsubscribe from the list and majordomo tells you he can't find your name on the list, send the "who spm" or "who spm-diges= t" command to find out who is on the list. Scan the addresses and search for your email address. Then send an unsubscribe command with that email=
address in it.
Please note that when you reply directly to a message, you are probably replying directly to the list. You will have to change the To: if you only want to reply to the author.
If your site starts bouncing mail back to me and it persists for several days, I will make no bones about deleting you from the list. You will then get a reminder sent every day that you have been bounced. That reminder will have instructions as to how you can get resubscribed again.=
I will have an archive of the old messages. They are available using the following ftp account at ftp.di.com. username: afm password: stm
Just to add my own $0.02 worth here, I have also experienced this problem. At that time I was infiltrating plant tissue, so it needed a long time to infiltrate because of cell walls, but the LR White seemed very non-viscous (what IS the word for non-viscous, I'm always looking for that word), very much like water, and then, with no accelerator it suddenly polymerized in the vials and surprized me and caused no end of grief because of that surprize. I thought that it would slowly increase in viscosity, such that I would be able to predict when it was going to polymerize, and this is what fooled me. It behaved quite differently from other resins that I've used in the past.
This was at least 6 years ago, so I cannot remember now if I had osmicated that tissue or not.
Garry
} ---------- } From: SOBOCIG[SMTP:sobocig-at-aa.wl.com] } Sent: 22 August, 1997 07:04 } To: Ann-Fook Yang; CarbynS-at-em.agr.ca; Microscopy-at-Sparc5.Microscopy.Com } Subject: LR White Question- Premature Polymerization. } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The Motorola SCG Chemical and Surface Analysis Lab is a multidiscipline analytical lab for wafer fab manufacturing in Phoenix, AZ. We are currently in need of a technician to provide TEM sample preparation support to our TEM operation. The candidate should have the following qualification. 1. Understanding of basic TEM imaging. 2. Experience or training in the following TEM sample preparation techniques: plane-view and cross-section TEM sample preparation using wedge polishing, dimpling; specific area cross-section with FIB. 3. Basic understanding of semiconductor devices and processing is highly desirable, but not required. 4. Applicants should have a A.A. degree in analytical technology, process good communication skill, and be able to work in a team-oriented environment.
Interested candidates should send resume directly to
Rebecca Ai MD P004 52nd St. Chemical and Surface Analysis Lab Semiconductor Component Group Motorola, Inc. 5005 E. McDowell Road Phoenix, AZ 85008 Ph. (602)-244-5775 Fax. (602)-244-6492 Email: RP3478-at-email.sps.mot.com
Bob: I have not found a listserver for AFM, but I have not looked for one for a while. You could check the AFM/Tunnelling section of the links on our site. There is a dozen good links, they may help anyway and perhaps point to the listserver - if one exists. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 400+ Links, MSDS ************************ http://www.proscitech.com.au
} Gentle folk, } } I believe that there has been mention of an atomic force microscopy } listserver at some time on this list. I am at home, recovering from surgery } and can't look through my filed mail at the office. } } We have begun doing some AFM work and I would like the address of } such a server, if it does exist. I'd like to make some good use of my } convalescence and learn more about the AFM before I return for work. } }
I have run into a couple of occassions of premature polumerization in the 10 or more years we have used LR White resin. In both cases the resin was old, ie over 1 year but was not associated with osmication. An possible explanation was suggested to me by Roy Gillett of London Resin a number of years ago who definitely recomended a shelf life of 12 months for catalysed resin.
Quote "The reason this pre-polymerisation occurs only with tissue must be something to do with a tissu constituent catalysing polymerisation. Older resin is much more susceptibe to this that fresh monomer becaue of the significant polymer growth that will inevitably have occurred in the monomer. The most likely 'endogenousd catalyst' from previous experience is likely to be an amine or peroxide moiety in the tissue"
We have had no problems since switching to buying uncatalysed resin and making up a new bottle as we run out of the old.
One point I have noticed in the discussions to date is some ambiguity between calalyst and accelerator. From my understanding the catalyst (benzoyl peroxide powder) must be added 24 hours before you start using a batch of resin and is necessary for both thermal (oven) and "cold" polymerization. The accelerator on the other hand is added to the final resin change for rapid "cold" polymerization without using an oven.
Ian
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-849 3660 EMail ihallett-at-hort.cri.nz
I have to embed some Drosophila melanogaster heads for transmission electron microscopy and prepare the same for scanning electron microscopy. The person requesting the work is interested in eye morphology. I have done a literature search and have come up with some relevant papers but my problem is that I will probably get these heads before I get the papers!! Working in the medical field, I have never embedded a sample with an exoskeleton before and am wondering if someone out there can give me an idea on the appropriate fixatives to use including fixation times and any little tricks I may need to know. Do the heads sink in the fixative or do I need to spin them down or use a vacuum? Is one resin better than another? How do they cut? Also, for SEM, which fixative do I use and for what duration? Again, are there any tricks to the dehydration and critical point drying procedures?
We have a TEM Siemens Elmiskop 101 in our lab. Recently, we encountered some problems with the high voltage controller. The cause was the bad connection of a K81A-type diode (electron tube-type diode) located in the power supply cabinet. We are looking for any K81A-type diode which could replace our old one.
Thank you for your help
Solvay Research and Technology Philippe Drouillon Rue de Ransbeek, 310 1120 Brussels (Belgium) Tel : (00 32) 2 264 24 47 Fax : (00 32) 2 264 20 55 Username : Philippe.Drouillon-at-solvay.com
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear colleagues,
Herewith I would like to thank, also on behalf of the other members of the "working group on accreditation of microscopical work" of the Dutch Society for Microscopy, all contributors and participants in the discussion on:
accreditation, callibration, standards, and ISO 9001.
CAPSULE DESCRIPTION OF JOB Has primary responsibility for developing a state-of-the-science Microsco= py & =
Image Analysis program for Nabisco in alignment with the Research Strateg= y. =
Proactively identifies, communicates, and executes key areas in microscop= y =
and image analysis. Participates on project teams in both technical =
leadership and support roles depending on the needs of the project. Prov= ides =
proactive support to co-workers in the area of microscopy and image analy= sis =
in both identifying how the technology can be effectively used in project= s =
and providing user-friendly microscopy setups for co-workers. Has comman= d of =
food material science/physical chemistry which enables understanding and =
effective partnering in cross-functional teams/efforts.
BASIC REQUIREMENTS =B7 Phd and 2-5 years experience; MS and 5+ years experience; BS and 7+ y= ears =
experience =B7 Degree in Food Material Science, Food Physical Chemistry, or related = field =B7 Develop a state-of-the-science microscopy & image analysis program fo= r =
Nabisco. In alignment with the Research Strategy, researches, identifies= , =
proposes and executes technology areas where microscopy can impact and ad= d =
value to the business and should be developed in the future =B7 Accesses current equipment and methodology and maximizes its use in t= he =
company. If other instrumentation/ methodology is required that is not =
currently in-house, develops a database using outside sources =B7 Expertise in light (bright field, polarizing & fluorescence) and elec= tron =
microscopy and image analysis =B7 Knowledge of food physical chemistry, material science, texture =B7 Excellent communication and interpersonal skills =B7 Creative, proactive and self-starter =B7 Interest and ability to work on cross-functional teams
-------------------------------------------------------- Dear Dr. Reffner,
We are unable to find any reference to the Microscopy list server in the = MSA =
website. We would appreciate it if you could therefore forward the attac= hed =
file for us. I am optimistic that you have the corresponding applicatio= n, =
and am therefore sending the file to you as an attachment:
If, however, you are unable to access the information, please either e-ma= il =
I've recently acquired a Oxford Instruments Link PCXA EDS X-Ray analyser - mid 1980s vintage but in good working order. My problem is the file format of the resulting spectra is markedly different to that from the contemporary QX2000/AN10000 analysers. I think its something to do with byte order being different for PC based processors. I need to be able to extract the files in order to label/print using ASCII readable spreadsheet software. If anyone has any experience/suggestions on the PCXA system, I'd be grateful to hear from them.
Thanks in advance,
Stu
****************************************************************** Stuart Kearns Electron Microbeam Laboratories Dept. of Geology tel: 44 (0)117 928 8204 University of Bristol fax: 44 (0)117 925 3385 Bristol BS8 1RJ UK email: Stuart.Kearns-at-bris.ac.uk ******************************************************************
Hello, my name is Lou Solebello and I am a research chemist/light microscopist with the JM Huber Company, Engineered Minerals Division in Macon Georgia. I am a newbie to SEM, and have not tried the Mquant software. A colleague of mine however, has been experiencing a lot of difficulty with getting the software to work properly. Does any one out there have experience and tips they would like to share? any help would be appreciated. I can be contacted by e-mail at either of the two addresses below.
First, for the TEM part: if you can get newly emerged adults, the cuticle will be soft and reasonably easy to section. By "newly emerged" I mean within a few hours. After the heads have assumed their normal shape (they inflate to rupture the pupal exoskeleton), but before they have tanned significantly. This assumes that the client isn't investigating something in the eye that changes subtly with tanning, or age after emergence.
The most difficult part of the eye for TEM are the crystalline cones. If the client isn't interested in them for TEM, dissect away the exoskeleton (cuticular area over the eye) and the underlaying c. cones. If possible, before fixation or embedding. (Waiting for the laughter to die here.) The easiest way to do this is chopping away with a glass knife after the blocks are ready.
You can handle the retinal tissues pretty much like any neural tissue. The client should already have the relevant fixation references, but routine fixation should be pretty normal. I used pH 7.2, 0.1-0.15 M buffers for freshwater crustaceans, standard Karnovsky's.
If s/he does want the cuticular and crystalline lenses ... wait for the references, or a response from a _Drosophila_ / small insect specialist--I worked on crustaceans (there are differences in the cuticle).
Use a *hard* resin with low viscosity. Other than that, I've seen no advantage for one type over another; except what works for you.
The heads should sink, if they don't, a *little* spinning won't hurt. By hand, I wouldn't use a centrifuge for this, even on the lowest speed. Mild vacuum would be better.
You mention "heads", so: is that what the client is bringing you, or are you planning on decapitating the critters? This would be good, if you can find a guillotine small enough. Best would be to then bisect the heads midsaggitally, if you can handle (orient, etc.) the resulting hemiheads. They may be pretty difficult to see to orient after OsO4, although the cuticle won't take up much osmium.
For SEM: treat like for TEM, except leaving the heads intact. CPD works well, and drying from HMDS can also (HMDS was originally used for Malphigian tubules in insects). Crystalline cones can be exposed nicely for SEM by dry-fracturing the heads after drying and mounting on the stubs. Read: hitting the eye with a razor blade, then gently blowing away the debris with a duster. (Also, look in through the back of an intact head.)
Mount on double-stickey *carbon conductive* tape, then run a thin line of silver paint to the head. Touch the wet paint to a sacrificial area of the head (one you don't care about), and draw a ring of Ag paint around the head, as close as you can get without touching it.
All of this last is to insure maximum conductivity of the specimen. The setae will charge like buggers otherwise, and you'll have bright little hairs, and lose structural details of both the setae and surrounding cuticle. Make certain that the heads have both excellent mechanical and electrical contact to the stub.
Phil } I have to embed some Drosophila melanogaster heads for transmission } electron microscopy and prepare the same for scanning electron } microscopy. The person requesting the work is interested in eye } morphology. I have done a literature search and have come up with some } relevant papers but my problem is that I will probably get these heads } before I get the papers!! Working in the medical field, I have never } embedded a sample with an exoskeleton before and am wondering if someone } out there can give me an idea on the appropriate fixatives to use } including fixation times and any little tricks I may need to know. Do } the heads sink in the fixative or do I need to spin them down or use a } vacuum? Is one resin better than another? How do they cut? Also, for } SEM, which fixative do I use and for what duration? Again, are there } any tricks to the dehydration and critical point drying procedures? } } Thanks in advance } } Sarah Ellis
I have to dry some bacteria grown on glass coverslips shortly for SEM from acetone to liquid CO2 i.e. critical point drying. I would appreciate advice in the next 24 hours about what times are recommended in the drier.
Unfortunately, the 'customer' has used quite a few 22 x 22 mm coverslips and I only have the original Polaron drier, without any specially-made coverslip holders, so it looks like three will fit in gauze baskets which I use for baby squid.
I spend my time embedding triatomino's legs. This is my method:
1)cut the heads 2)put 2-3 hs. in fixative medium ( I don't use transmition mic.) 3)deshidratation: ROH 70% (1*10 min) -- 80% ---- -- 90% ---- EtOH 100%(3*10 min) 4)EtOH 100% + Propilenoxide : (1:1) : ( 10-15 min) 5)Propilenoxide (10-15 min) 6)Propilenoxide + Durcupan (epoxi resine): (1:1) : ( 1 night) 8)open the recip. at 40 centigrade degrees ( 1h. 30 min) 9)put in new Durcupan at room temperature (1h.) 10)Put in new durcupan and orientation the head (1 night at 60 cent. degrees) Veronica Campanucci -------------------------------------------------------------------------------- Veronica Andrea Campanucci Laboratorio de Fisiologia de Insectos Dpto. de Ciencias Biologicas Facultad de Ciencias Exactas y Naturales Tel (54 1) 781-5021 to 29, ext. 332 Universidad de Buenos Aires FAX (54 1) 782-0582/544-7893 (1428) Buenos Aires e-mail: veronica-at-bg.fcen.uba.ar Argentina HTTP://biolo.bg.fcen.uba.ar/physinse.htm --------------------------------------------------------------------------------
I am looking for any suggestions, advice with ongoing problem with weak immunostaining of semithin sections (0.5-1 micron) of low temperature Lowicryl K4M-embedded infiltrating ductal breast carcinoma.
After successful experiments on paraffin sections of the tissue, an incubation protocol for cytokeratin-8 antigen detection with immunoperoxidase-DAB procedure was applied to Lowicryl semithin sections, without the removal of the hydrophilic resin. The semithin sections were reacted with the antibody overnight at 4oC, and negative controls were performed by substituting the primary antibody with PBS. An increase of the staining contrast was obtained by posttreatment with OsO4. No staining was observed, as expected, at semithin sections used as controls. However, a very weak yellowish staining was observed at the sections treated with the which could not be evaluated. Trying to increase the intensity of the immunoreaction, I increased the concentration of the second Ab, the thickness of the sections (3-4 microns), and the incubation time of OsO4. The modifications took place at different experiments, but none of them was effective.
What should I do?? Any help would be greatly appreciated. Thank you in advance.
} ---------------------------------------------------------------------------- } Welcome to } } Microscopy-at-MSA.Microscopy.Com } ---------------------------------------------------------------------------- } } Hosted and Sponsored by The Microscopy Society of America (MSA) } as a Free Service to the World-Wide Microscopy & Microanalysis Community } ---------------------------------------------------------------------------- } Your subscription request to the Listserver/MailReflector has been received. } Please reply to this message to confirm that your address has been } successfully recorded. When your reply is received your address will be } added to the Mailing List. } } ********************************** } IMPORTANT } please send this reply to } LISTSERVER-at-MSA.Microscopy.Com } ------------------------------------------------ } DONOT send it to Microscopy-at-MSA.Microscopy.Com } otherwise everyone on the server will } see your message and this creates unnecessary } and annoying traffic!!!!!!!!!!!! } ************************************* } } The purpose of this reply is to insure that no typographical errors have } occured during your subscription to the system and to minimize the } transmission of Email to incorrect addresses which sometimes result in } messages being bounced/reflected to a large list of receipents tying up the } system (and your mail account!). } } After sending in your reply all discussion/comments/questions which you } would like to be distributed to the list should be done by sending the text } as a simple Email message } } To: Microscopy-at-MSA.Microscopy.Com } } All subscribers to the list (including yourself!) will automatically receive } copies of the message you post. } } As a courtesy to the readers of this list please indicate in the subject } line of your message a reasonable descriptive title of your comment/inquiry. } Also preface your description by the conventional abbreviation of the type } of microscopy you are interested such as LM, IRM, XRM, TEM, SEM, AFM, STM, } uProbe...... For example, if you are interested in optical microscopy and } have a question about staining then a Title/Subject line for your message } might be } } Subject: LM - Need help on Staining Cells } } or if you are interested in TEM analysis of dislocations then } } Subject: TEM - Dislocation Loop Analysis } and so forth. } } Address all problems, questions, subscriptions, confirmation replies, and } unsubscriptions to: } } Listserver-at-MSA.Microscopy.Com } } Thanks for your Cooperation: } } Nestor J. Zaluzec } Email:Zaluzec-at-MSA.Microscopy.Com } Tel: 630-739-1160 } } =========================================================================== } } ---------------------------------------------------------------------------- } } General Ground Rules on using the MSA Microscopy Listserver/Mailreflector } } ---------------------------------------------------------------------------- } Before continuing you should understand your responsibilities as a } Microscopy Listserver/Mailreflector System user. } } Specifically they are: } } * 1.) Actively encourage and promote the free exchange and discussion of } information, ideas and opinions, except when the content would } compromise the national security of the United States (or any other } country for that matter) ; violate proprietary rights, personal } privacy, or applicable state/federal/local laws and regulations } affecting telecommunications; or constitute a crime or libel. } } * 2.) Use your REAL NAME and fully disclose any personal, financial, or } commercial interest when evaluating any specific product or service, or } contribution. } } * 3.) Do not use this system for delivery of personal mail, messages or } items of a similiar nature (i.e no posting of resume's etc....) If you } are not sure then it probably does not belong here! If you would like } an opinion then Email the message to me and I will review and comment } as appropriate. (Zaluzec-at-aaem.amc.anl.gov). } } * 4.) Adhere to these rules and notify the Listserver-at-aaem.amc.anl.gov } immediately when you discover any violations. } } ======================================================= } Additional Information } ======================================================= } } Anonymous FTP access to files in the Microscopy and Microanalysis Software } Library (MMSLib), and the Listserver Archives. } } FTP SITE ADDRESS : ftp.msa.microscopy.com } UserName : Anonymous } Password : Your Email Address } WWW SITE ADDRESS : http://www.msa.microscopy.com } ======================================================= } Now for some Legal stuff } ======================================================= } } Every user explicitly acknowledges that all information obtained from this } system is provided "as is" without warranty of any kind, either expressed or } implied, including, but not limited to the implied warranties of } merchantability and fitness for a particular purpose and that the entire } risk of acting on information obtained from this system, including the } entire costs of all necessary remedies, is with those who choose to act on } such information and not with the operators of this system or the suppliers } of the information. } } Email: Nestor J. Zaluzec } } } ========================================================================== } To: NewSub-at-MSA.Microscopy.Com } From: "Nestor J. Zaluzec" {zaluzec-at-microscopy.com} } Subject: FAQ File - Microscopy ListServer } Cc: } Bcc: } X-Attachments: } } ---------------------------------------------------------------------------- } Help/FAQ File } THERE IS ALOT OF IMPORTANT INFORMATION HERE. } MAKE SURE YOU READ IT ALL.... } ----------------------------------------------------------------------------- } ************************************** } FAQ's File } Microscopy/Microanalysis Listserver: } ************************************** } Last Updated April 15, 1997 } ************************************** } } ---------------------------------------------------------------------------- } What is the purpose of this Listserver? } ---------------------------------------------------------------------------- } } The purpose of this system is to give the scientific community a centralized } Internet (Email) address to which questions/comments/answers in the various } fields of Microscopy & Microanalysis can be rapidly distributed to a list of } interested individuals and organizations. } } ---------------------------------------------------------------------------- } What type of Microscopy/Microanalysis does this List address? } ---------------------------------------------------------------------------- } } For the purposes of this discussion forum, Microscopy & Microanalysis should } be considered to include all techniques which employ a probe such as: } photons (including xrays), electrons, ions, mechanical and/or } electromagnetic radiation to form a representation or characterization of } the morphology, crystallography, elemental, chemical or electronic structure } of any material in either physical and/or life sciences applications. } } Some of the more common techniques which are associated with these fields } include the following: optical microscopy, xray microscopy, scanning } electron microscopy, transmission electron microscopy, atomic force } microscopy, scanning tunneling microscopy, scanning ion microscopy, } analytical electron microscopy, electron microprobe, xray energy dispersive } spectroscopy, electron energy loss spectroscopy, electron diffraction, } convergent beam diffraction, high resolution electron microscopy, high } voltage electron microscopy, etc. (the list continues...ad infinitum.. ) } } ---------------------------------------------------------------------------- } How do I subscribe? } ---------------------------------------------------------------------------- } } Send an Email Message to "Listserver-at-MSA.Microscopy.Com" in the body of your } message include the lines } } Subscribe Microscopy "USERID-at-EMailAddress" Optional: Real Name } Optional: Real (Surface Mail) Address } Optional: Microscopy/Microanalysis Interests } } Replace the "USERID-at-EMailAddress" with your real Email address. You may also } append your Real Name, and Address etc.. to the end of the message as a } courtesy. I'd be interested to know who you are and where you're from, and } what you do. } } ---------------------------------------------------------------------------- } How do I unsubscribe? } ---------------------------------------------------------------------------- } } Send an Email Message to "Listserver-at-MSA.Microscopy.Com" in the body of your } message include the line } } Unsubscribe Microscopy "USERID-at-EMailAddress" } } Replace the "USERID-at-EMailAddress" with your real Email address. } } IMPORTANT NOTE#1. To unsubscribe you must supply the original address which } you used when you first submitted your subscription. Over the last few years } I have found that many users have their mail forwarded to new addresses, or } use an alias. When they then try to unsubscribe, they forget where there } mail is being sent to (or forwarded from). This causes me lots of headaches. } So just remember in order to unsubscribe you from the list, the listserver } must know the your original subscription address to remove it! } } Please be considerate. I am not a mindreader, and don't really enjoy playing } detective over the Internet. I have too many other things to do. } } IMPORTANT NOTE #2: Do not send subscribe/unsubscribe message to } "Microscopy-at-MSA.Microscopy.Com". When you do this ALL members of the } listserver receive your message. All administrative issues should be sent to } "Listserver-at-MSA.Microscopy.Com" and not to the general mailing list. } } ---------------------------------------------------------------------------- } Are There Any Special Settings on my Email Program? } ---------------------------------------------------------------------------- } } Yes. } } 1.) Please insure that your correct Email address is listed in the FROM: } record. You may need to enter this in the Preferences option of your Email } program. Contact your system administrator if your not sure what to do here. } } 2.) DONOT set your Email Program to automatically reply to all messages, or } to request a return receipt. If either of these are done, you run the risk } of creating an Email loop within the system. This may result in a message } bouncing through the system for several days, filling up everyone's mail } box! } } 3.) Generally it is best to terminate each line of your message at less than } 80 characters. This avoids the occassional problem of messages getting } chopped up. Some mail routers, limit the number of characters per line. If } you exceed 80 characters it is sometimes possible that your message will } become randomly truncated. If your not sure, then simply enter a (Carriage } Return) at the end of each line of text you type (ie. don't let your system } automatically word wrap) and things should be fine. } } ---------------------------------------------------------------------------- } How do I submit/post a message? } ---------------------------------------------------------------------------- } } Compose your message offline, then read it a second time! } } Send your message by Email to: "Microscopy-at-MSA.Microscopy.Com" } } As a courtesy to the readers of this list please indicate in the subject } line of your message a reasonable descriptive title of your comment/inquiry. } Also preface your description by the conventional abbreviation of the type } of microscopy you are interested such as LM, IRM, XRM, TEM, SEM, AFM, STM, } uProbe...... For example, if you are interested in optical microscopy and } have a question about staining then a Title/Subject line for your message } might be } } Subject: LM Need help on Staining Cells } } or if you are interested in TEM analysis of dislocations then } } Subject: TEM Dislocation Loop Analysis } } This way individuals can filter out the messages that they may not be } interested in. } } Can I send an Email Attachment? } } Please DO NOT send files as an attachment to the Listserver. } } Many people use email software which displays plain text messages } automatically but requires several manual steps to read and delete or save } an attachment Others have no capabilities at all for extracting and or } translating attachments. Please send all messages as plain text. This will } insure that ALL subscribers to the Listserver can rea ALL messages... } } ---------------------------------------------------------------------------- } How do I reply to a message: } ---------------------------------------------------------------------------- } } Compose your message offline, then read it a second time! } } Send a NEW message to "Microscopy-at-MSA.Microscopy.Com" } } DONOT use the REPLY function of your Email Program!! } } If you do this (i.e. use REPLY) then, in most cases, only the person that } originated the message will see your response. The current configuration of } the Listserver is such that a message is basically REFLECTED from the } MSA.Microscopy.Com site to all subscribers on this list. the repackaging of } the message is minimal and not all Email programs will recognize that the } message reply needs to go back to Microscopy. A good fraction will reply } back only to the originator. Therefore, if you want everyone to benefit from } your problem and and/or it's answer you must send it as a message to } "Microscopy-at-MSA.Microscopy.Com" } } ---------------------------------------------------------------------------- } What can I post? } ---------------------------------------------------------------------------- } } Basically any question/comment/observation or general } information/announcement which involves microscopy and/or microanalysis. } This list is not moderated, so it is up to the users to selfpolice } themselves. I generally take an openminded attitude toward questions. If in } my opinion, it appears that a discussion is straying far off the limits of } the intent of this forum I will post a note to the group. But I prefer to } err on the safe side with most messages and allow at least a modicum of } generally. Clearly, generic questions about Word Processing do not belong in } this discussion forum, so just use common sense. } } If there is any question in your mind about something you wish to post, send } it to me first at Zaluzec-at-MSA.Microscopy.Com and I will give you my } opinion/comments. } } ---------------------------------------------------------------------------- } Can I post an Announcement of a Job Opening or a Meeting? } ---------------------------------------------------------------------------- } } Yes that falls within the bounds of the subject of this list as long as it } is related to Microscopy/Microanalysis. } } ---------------------------------------------------------------------------- } Can I post my Resume'? } ---------------------------------------------------------------------------- } } No. This forum was not created for that purpose. } } ---------------------------------------------------------------------------- } Can I post an Advertisement? } ---------------------------------------------------------------------------- } } No, that does not fit within the bounds of this discussion forum. } } This listserver is not intended to be a Sales mechanism for commerical } organizations, but rather it is an open discussion area about microscopy and } microanalysis problems and solutions. If you are an organization and have } equipment you wish to donate, or sell, for nominal cost (i.e. no profit) } then this is generally an acceptable posting. If you are not sure then send } a copy of the announcement in question to Zaluzec-at-MSA.Microscopy.Com and I } will give you my opinion. An example of this type would be an old } decommissioned instrument which someone is trying to give away for } removal/shipping costs, that would fit within the bounds of the purposes of } this list. } } If you are a manufacturer, you are always welcome to observe/join in any } discussion at any times. We do ask that everyone, please refrain from overt } sales pitches and/or commericalism. If a product which you produce/sell can } solve a problem or answer a question raised by anyone on this list, then by } all means feel free to say so. Try to be brief about the product, state the } simple facts in a few (short) sentences and then offer to continue the } discussion with any interested parties offline. Alternatively you can give } sufficient information so that individuals can download/access the relevant } information.Usually it will be sufficient to just add your phone number } and/or Email address to the end of your message, and you'll be contacted by } anyone that is interested. } } Remember, please keep your comments about any product you "sell" to a } minimum. } } It is not out of line to provide your company name, Email address or WWW } site as part of your signoff/signature line, at the end of ANY message you } post to this system. } } This Listserver operates on the honor system with respect to to posting of } advertising, so please respect these simple ground rules. } } If you are interested in using the Internet for Commerical Advertising of } Microscopy/Microanalysis Related Products/Services, you may wish to contact } MSA at it's WWW site (http://www.msa.microscopy.com) or the MSA Business } Office (MSABusinessOffice-at-MSA.Microscopy.Com). These alternative Internet } services, are provided independently of the Listserver Operation, which MSA } provides as a FREE service to the WorldWide Microscopy and Microanalysis } Community. Any funds derived from the above are used to defray the costs of } running MSA's Internet site. } } ---------------------------------------------------------------------------- } I'm only interested in XYZ Microscopy is there sublist? } ---------------------------------------------------------------------------- } } Nope, not here. There are other listservers which deal with specific topics, } but this one is general. At sometime in the future I may consider creating } subtopics. But really that is what the SUBJECT line is for. Please use a } good description for your message! I've learned alot by just "listening" in } on discussions about topics I know nothing about. You may also find the same } is true for you. } } ---------------------------------------------------------------------------- } Can I access this list via NewsGroups on the Internet? } ---------------------------------------------------------------------------- } } Not any longer. For a time this Listserver automatically forwarded all } postings to the SciTechnqiue.Microscopy Newsgroup. However, the link to that } newsgroup has been lost at the receiving end. A link may once again be } possible in the future, when hardware and software is upgraded. } } ---------------------------------------------------------------------------- } I think I need to test the Email, what do I do? } ---------------------------------------------------------------------------- } } First, if you received the instructions, after submitting instructions, then } your Email program must be working. So no further testing is really } necessary. } } DO NOT send TEST/HELLO messages to Microscopy-at-MSA.Microscopy.Com it } needlessly clutters up the subscribers mailboxes. } } Remember every time you post a message to the Listserver over 2000 copies } are sent. This not only wastes processing time on my computer, but gets lots } of people angry at wasted bandwidth. You should also remember some people } have to pay $$ just to read their Email (think of those subscribers on } CompuServe, AOL, GENIE, etc....) We have a lot of them, and this wastes time } &/or money! } } If you really feel you must test your Email Package or think you need help } then send a message to: } } Zaluzec-at-MSA.Microscopy.Com. } } He's sometimes a patient soul, who at times has been known to read his Email } and occasionally answers questions. Especially, late at night. } } ---------------------------------------------------------------------------- } Is there a Digest Mode? } ---------------------------------------------------------------------------- } } Digest Mode is not available on the present server. It is an option which } will be added in the future. } } Digest Mode compresses all message from a single day into one long message } which is then sent to all subscribers. It is a nice option, and I intend to } get it running eventually. } } ---------------------------------------------------------------------------- } When I send a message I get (sometime many) Bad/Bounced Messages back why? } ---------------------------------------------------------------------------- } } Because this list acts as a reflector, when message delivery fails and is } returned to the system it goes to the originator of the message. Due to the } large number of users on this list it is ineveitable that some systems will } go offline, are down for maintenance, or simply shut down by the user. The } mailserver tries to deliver messages several times before it gives up. } } ---------------------------------------------------------------------------- } Can I get a list of the Subscribers: } ---------------------------------------------------------------------------- } } No. I do not release that information. There are in excess of 3000 } subscribers from more than 40 countries to this list. } } ---------------------------------------------------------------------------- } Is there an Archive? } ---------------------------------------------------------------------------- } } Yes. You can access the monthly archive at the WWW site: } http://www.msa.microscopy.com/MicroscopyListserver/MicroscopyArchives.html } } ---------------------------------------------------------------------------- } What is the subscription fee? } ---------------------------------------------------------------------------- } } The Listserver is a free service brought to you by MSA there are no charges } for subscription/use. I'd even be willing to guarentee that you will get } back at least whatever you put in. If you feel generous, and you happen to } see your Friendly Neighborhood SysOp at a Microscopy Conference you can } always buy him a beer or two. He's usually easy to recognize, and his arm } twists easily. } } ---------------------------------------------------------------------------- } Who do I contact with problems? } ---------------------------------------------------------------------------- } } God-at-Garden.Eden.Earth.Com } } ---------------------------------------------------------------------------- } Who Runs this? } ---------------------------------------------------------------------------- } } This listserver is run by Nestor J. Zaluzec (Zaluzec-at-MSA.Microscopy.Com) It } has been & continues to be run/administered/babied mostly in his spare time } and usually late in the evening or earlyhours of the morning. } } ---------------------------------------------------------------------------- } Special Announcement/News } ---------------------------------------------------------------------------- } } At the 1995 Winter Council Meeting, the Microscopy Society of America (MSA) } approved a proposal to support this server as well as other } telecommunications options as a FREE Service to ALL persons/organizations } worldwide who are involved or interested in Microscopy and/or Microanalysis. } } Thanks to this support, the listserver will be upgrading the hardware } software and services for use by Microscopist's/Microanalysts Worldwide. } } ---------------------------------------------------------------------------- } Is there an FTP and/or WWW site? } ---------------------------------------------------------------------------- } } Yes, both an FTP and WWW sites exist here are the addresses: } } WWW http://www.msa.microscopy.com } } FTP ftp.msa.microscopy.com (Anonymous Login Enabled) } } ********************************* } Last Updated April 15, 1997 } Nestor J. Zaluzec } Your Friendly Neighborhood SysOp. } ********************************* } End of File } ********************************* } } }
Anyone know of a review article or bibliography on cathodoluminescence? I have one dating back to 1977 but I imagine there must be a more recent one. I did a lit search but came up negative. Any help would be appreciated. Thanks.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 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 ####################################################################
1. What is your experience using field emission SEM with EDXS.
2. What are the advantages and disadvantages. Is there sufficient beam current?
3. Would we be better off using tungsten filament source?, LAB6 source? for EDXS?
4. Imaging is also very important. As a reference point, we have a JEOL 6300F and are very pleased with the imaging at low keV. Are the current tungsten or LAB6 source microscopes being sold capable of providing the same level of image quality as a FE source microscope in the 1-5 keV range?
I really appreciate your time spent responding to these questions, however briefly.
There are many protocols that work. My experience indicates that 2% glut, 4% form in .1M PO4 works well for most TEM & SEM. A few minutes centrifugation at low speed (200rpm) hasn't damaged my samples. It helps to cut the proboscis off if you are not allowed to bisect the head. Standard EmBed resin has worked well for me--just allow infiltration overnight. Proper orientation can be the most difficult often requiring two or more embeddings, reorienting each time. SEM prep is very similar though I have found that critical point drying may require extended CO2 soaking times (30 to 60 minutes X 3). Some of our mutants have very little tissue in the head and are very susceptible to drying artifacts (they collapse). Some people leave the head attached to the body for ease of handling. I don't. Use low accelerating voltages in the SEM (10 kV or less) to minimize charging, etc. Good luck! Larry D. Ackerman (415) 476-8751 Howard Hughes Medical Institute FAX (415) 476-5774 UCSF, Box 0724, Rm U426 533 Parnassus Ave. mishot-at-itsa.ucsf.edu San Francisco, CA 94143
} ... } 1. What is your experience using field emission SEM with EDXS.
... I shopped for a like instrument in '92 ...
} 2. What are the advantages and disadvantages. Is there sufficient } beam current?
Primarily your concerns for FE should be beam stability... enuf beam current was good question at the time ... but most FE guns were capable of delivering several nanonamps ... enuf for EDX.
}
} 3. Would we be better off using tungsten filament source?, LAB6 } source? for EDXS?
... better off, yes (... more stable ...), but you wouldn't get the low keV performance you mention below.
} 4. Imaging is also very important. As a reference point, we have a } JEOL 6300F and are very pleased with the imaging at low keV. Are the } current tungsten or LAB6 source microscopes being sold capable of } providing the same level of image quality as a FE source microscope in } the 1-5 keV range?
... you could come close with a LaB6 ... or a well designed W gun, but I still believe you'd find yourself optimizing the gun for low keV performance by varying the cathode-tip/wehnelt distance ... not something you want to be doing every day. Lastly, a FE gun is not likely going to provide you with the beam current stability you may want for EDX, especially for element mapping. I do remember there being methods of stabilizing a FE gun via feedback, but you want to make sure that the feedback isn't simply stabilizing the image contrast/brightness, but rather the beam current itself.
cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
} } 1. What is your experience using field emission SEM with EDXS.
3.5 years. AMRAY 1845FE w/Noran Voyager III, thin window. Plug and play. I work in an industrial services lab supporting many JIT manufacturing plants. Uptime is critical and the system has delivered as needed. The system has paid for itself over and over again. If I had the money, I'd upgrade all of our scopes to FE. } 2. What are the advantages and disadvantages...
Sufficient current has never been a problem. 18 nanoamps w/o apertures. We can easily swamp the detector with 100 micron aperture that provides a good balance between x-ray production and image quality. } 3. Would we be better off using tungsten filament source... We also have a highly modified AMRAY 1600 upgraded to a LaB6 last year. The scope is now more of a probe than an imaging tool. Light element EDS, 4 crystal WDS, CL, BSED, air-lock with sputtering gun, etc. Vinnie Casasanta, now at Charles Evans & Associates (Surface Sciences) built it specifically to support our materials development labs. Gun operation no problem. Just had to find the "sweet spot" in the saturation curve. Actually, Vinnie got enough current when he used an output valley rather than a peak because it provided a more stable beam. } } 4. Imaging is also very important...
I regularly use 500V to 1.5 keV for imaging at { { about 10k mag. At 5 keV, 50 thousand plus, no problem. With the LaB6 SEM, the instrument isn't tuned for imaging, it is just a directional current source so I can't really compare apples to apples.
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 Phone: (508) 750-1717 E-mail: crossman-at-osi.sylvania.com
Our web sites: www.sylvania.com www.siemens.com --
dear colleagues... a while ago, there was some chit-chat about a microscope from a company called Tasco...located in the northwest, I think... of course, now that i need the info i can't find it....I'd be grateful if someone could give me a lead to help me locate them... thank you... Ron Mervis ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Ronald F. Mervis, Ph.D. Chief Scientific Officer Neuro-Cognitive Research Laboratories Columbus, Ohio
} Lastly, a FE gun is not likely going to provide you with the beam } current stability you may want for EDX, especially for element mapping.
True, but I would imagine that any modern EDX system (ours included) provides some mechanism for normalizing maps for beam current variations. So don't worry about this one too much.
Regards, Rick Mott, PGT rick-at-pgt.com www.pgt.com
} remember there being methods of stabilizing a FE gun via feedback, but you wa } nt } to make sure that the feedback isn't simply stabilizing the image } contrast/brightness, but rather the beam current itself. }
I saw a paper a few years ago written jointly by someone from Hitachi and someone from Kevex, it described such a true feedback system, might be worth contacting either of those companies.
Ritchie
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
We agree with your assessment pertaining to the one year shelf life for catalysed LR White resin under ideal conditions. It is one of the reasons that we began shipping the uncatalysed resin several years ago. We will, if a researcher needs it, catalyse it and send it to them.
We also are in complete agreement on your last point about the catalyst and the accelerator.The catalyst is needed for both "hot and "cold" polymerisation, while the accelerator is added only for the rapid "cold" polymerization.
I have a Denton 502A vacuum evaporator that doesn't suck like it used to. I've tried looking at it and I'm getting pretty frustrated. The problem is it doesn't ROUGH Pump like it used to, it acts like it has a leak. Does anyone out there know of a company or individual who does service calls on Denton Vacuum evaporators that's on the west coast, preferrably in the San Francisco Bay Area? Please help me! I'm close to just hitting the thing with a hammer! Thanks for any information you might give me.
Frustrated in Berkeley,
Paula = ) psic-at-uclink4.berkeley.edu
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
First, do you *have* to CPD? I've used HMDS very successfully for bacteria, in some cases retaining the "slime" sheath (that depends more on dehydration). 5 min. changes, 100% Ac =} 1:1 Ac:HMDS =} 3 X 100% HMDS, dry at room temp or 60 C
Air drying straight from acetone can work for bacteria nicely--do you have time to experiment?
For CPD, use 3 to 5 changes of CO2, with 2 to 5 minutes soaking in CO2 between changes (time by slime coat). Your real problem is going to be turbulence on filling and emptying the chamber, so this will have to be done *very slowly* so has not to disturb the coverslips.
Note: this is Ed Basgal's design and idea!: A cover-slip holder can be quickly made by cutting notches in a polyethylene or glass tube (not tygon), just wider that the thickness of the coverslips, so the slips fit down into the notch:
_ |_ {--cover slip fitting into notch | | | |__|
place tube into a polyethylene syringe body or centrifuge tube that's been fenestrated by a mad perforator. Cap both ends.
Phil
} I have to dry some bacteria grown on glass coverslips shortly for SEM } from acetone to liquid CO2 i.e. critical point drying. I would appreciate } advice in the next 24 hours about what times are recommended in the } drier. } } Unfortunately, the 'customer' has used quite a few 22 x 22 mm } coverslips and I only have the original Polaron drier, without any } specially-made coverslip holders, so it looks like three will fit in gauze } baskets which I use for baby squid. } } Regards - Keith Ryan } Plymouth Marine Lab., UK
I spend my time embedding triatomino's legs. This is my method:
1)cut the heads 2)put 2-3 hs. in fixative medium ( I don't use transmition mic.) 3)deshidratation: ROH 70% (1*10 min) -- 80% ---- -- 90% ---- EtOH 100%(3*10 min) 4)EtOH 100% + Propilenoxide : (1:1) : ( 10-15 min) 5)Propilenoxide (10-15 min) 6)Propilenoxide + Durcupan (epoxi resine): (1:1) : ( 1 night) 8)open the recip. at 40 centigrade degrees ( 1h. 30 min) 9)put in new Durcupan at room temperature (1h.) 10)Put in new durcupan and orientation the head (1 night at 60 cent. degrees) Veronica Campanucci -------------------------------------------------------------------------------- Veronica Andrea Campanucci Laboratorio de Fisiologia de Insectos Dpto. de Ciencias Biologicas Facultad de Ciencias Exactas y Naturales Tel (54 1) 781-5021 to 29, ext. 332 Universidad de Buenos Aires FAX (54 1) 782-0582/544-7893 (1428) Buenos Aires e-mail: veronica-at-bg.fcen.uba.ar Argentina HTTP://biolo.bg.fcen.uba.ar/physinse.htm --------------------------------------------------------------------------------
Veronica Campanucci -------------------------------------------------------------------------------- Veronica Andrea Campanucci Laboratorio de Fisiologia de Insectos Dpto. de Ciencias Biologicas Facultad de Ciencias Exactas y Naturales Tel (54 1) 781-5021 to 29, ext. 332 Universidad de Buenos Aires FAX (54 1) 782-0582/544-7893 (1428) Buenos Aires e-mail: veronica-at-bg.fcen.uba.ar Argentina HTTP://biolo.bg.fcen.uba.ar/physinse.htm --------------------------------------------------------------------------------
you posted a query re cathodoluminescence which I accidentally deleted, I have an expert here, if you want to mail me direct I'll pass on your query for review article(s)
Apologies if I've misspelled your name
Ritchie
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
Subject; Charging for usage in the multiuser EM Unit
We are a university based multi-user EM Unit that is moving towards the 'real' world. Presently, all university users of our EM Unit are charged for the consumables they use (based on an honesty system) but charged for nothing else. Our bean counters are now requiring us to introduce equipment usage charges, labour charges and occupancy charges. At this point I have no idea how much they expect us to recover, the wording that is used is that those who use the service should contribute to the cost of running the service. Contribute is the key word.
I have no problem with this concept at all. My problem is however, how do we record the many 'chargables' and I am hoping that those of you out there who have introduced such charging systems to your Units and those who are working with such systems can help me.
Our EM Unit is multi-user Unit where 80% of the users come to the Unit and do the work themselves after we have trained them.
Equipment Charges Charging for electron microscope usage is simple enough taken just from a HT counter however how do you charge for the ultramicrotomes, microwave oven, tissue processor, pH meters, osmometer, enelargers, photo printer etc without having a lot of clipboards all over the place ?
Occupancy How do you keep records of all the users coming and goings from the Unit so that a sensible occupancy charge can be made ?
Labour Charges Easy to do by introducing a 'Job Card' system for the work we do.
All help and suggestions appreciated
Regards
Allan
----------------------------------------------------------------------- Richard Lander Electron Microscope Technician South Campus Electron Microscope Unit Otago School of Medical Sciences P.O. Box 913 Dunedin New Zealand. Tel. National 03 479 7301 Fax. National 03 479 7254
"Southernmost EM Unit in the World!" ------------------------------------------------------------------------
I have examined microorganisms in the SEM directly, in their hydrated state with greatest success. I have used sputter coated or uncoated powdery mildew and rust colonies, and have about 30min before the spores dehydrate; any other treatment disturbes the delicate structures. The minute amounts of free water involved have not caused any problems with the vacuum or contamination; you obviously want to be sensible about this. One thing to remember, however, is that you will see the cells as they are, slime and all, which may obstruct interesting features of the bacteria.
If you have time to experiment, it might be well worth trying the simplest approach.
best wishes
Stephan Helfer
Sincerely +----------------------------------------------------------------- |Dr Stephan Helfer, SSO |Senior Mycologist - MSc Course Director | |Royal Botanic Garden Edinburgh, Inverleith Row, EDINBURGH EH3 5LR, |Scotland UK | |http://www.rbge.org.uk | |phone: +44 (0)131 552 7171 ext 280 | or +44 (0)131 459 0446-280 (direct digital VoiceMail line) |fax: +44 (0)131 552 0382 +------------------------------------------------------------------
Have you checked the air leak valve ? We once had a similar problem with our DV502 and it turned out to be a small leak in that valve. We have since put a filter right where the air goes in to prevent dust particles from collecting and causing poor seals.
I have a Denton 502A vacuum evaporator that doesn't suck like it used to. I've tried looking at it and I'm getting pretty frustrated. The problem is it doesn't ROUGH Pump like it used to, it acts like it has a leak. Does anyone out there know of a company or individual who does service calls on Denton Vacuum evaporators that's on the west coast, preferrably in the San Francisco Bay Area? Please help me! I'm close to just hitting the thing with a hammer! Thanks for any information you might give me.
Frustrated in Berkeley,
Paula = ) psic-at-uclink4.berkeley.edu
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
Have a look at this book: B. G. Yacobi, D. B. Holt: Cathodoluminescence in inorganic solids. New York: Plenum 1990.
Dr. Hartmut S. Leipner Fachbereich Physik Friedemann-Bach-Platz 6 Martin-Luther-Universitat D-06108 Halle
Tel. +49-345-55 25 453 Web http://www.physik.uni-halle.de/Fachgruppen/Kristall/index.html
-----Original Message-----
Hello again, I have rec'd. a sample for TEM that may be infected by a herpes virus. I am wondering if there is a way of enhancing the tissue with any stains (tannic acid etc.) to optimally show the virus. The tissue has been fixed in 4%GA, cacodylate buffer. I will be processing today, so I appreciate any info asap. Sorry for the short notice. Thanks Linda Fox lfox1-at-wpo.it.luc.edu
A recent message contained a misprint. ** Alwyn Eades Center for Microanalysis of Materials University of Illinois at Urbana-Champaign Phone 217 333 8396 Fax 217 244 2278 eades-at-uimrl7.mrl.uiuc.edu (NB those are letter l not ones) **
------------------------------------------------------------------------=20 The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20 =20 Hello All! =20 =20 I have a Denton 502A vacuum evaporator that doesn't suck like it used to=2E I've tried looking at it and I'm getting pretty frustrated=2E =20= The=20 problem is it doesn't ROUGH Pump like it used to, it acts like it has a=20 leak=2E Does anyone out there know of a company or individual who does service calls on Denton Vacuum evaporators that's on the west coast,=20 preferrably in the San Francisco Bay Area? Please help me! I'm close to just hitting the thing with a hammer!= =20 Thanks for any information you might give me=2E =20 =20 Frustrated in Berkeley, =20 =20 Paula =3D ) psic-at-uclink4=2Eberkeley=2Eedu =20 Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4=2Eberkeley=2Eedu =20 =20 =20 Paula; =20 Denton evaporators are almost indestructable, even hitting them with a= =20 hammer will not take them out of action for long=2E =20 The roughing and backing valves each have a bellows assembly inside=20 the valve housing, over time (several years) the bellows can develop=20 cracks resulting in leaks and long or impossible pump downs=2E This i= s=20 a likely possibility, but check the following first=2E =20 Do you have normal foreline pressure when both backing and roughing=20 valves are closed? This pressure should be 20mT or less=2E =20 =20 If this pressure is ok then what is the pressure when the backing=20 valve is opened? This too should be 20mT after a few minutes=2E =20 What is the best roughing pressure that you can achieve in 1, 5, and=20 10 minutes=2E =20 =20 If the foreline pressure and DP backing pressure are good but the=20 roughing pressure is poor, the main valve seal could be bad or the=20 seal around the bell jar could be poor=2E The bell jar could even have= =20 chips or hairline cracks=2E =20 =20 What is the best roughing pressure that you can get? If you can get=20 to high vacuum pumping what is the best pressure that you can get on=20 the penning gage? =20 Has the system ever been operated with both the backing and roughing=20 valves on at the same time, this is a bad thing! This will require=20 removal of the DP and inspection of the oil (color and amount)=2E =20 Usually when the backing and roughing valves have been opened=20 together, the DP stack will be displace upward resulting in poor high=20 vacuum performance or may even be impossible to hi vac pump=2E =20 Do you have manual roughing and backing valves (hand operated) or a=20 pneumatic system? Both of these use the bellows I described earlier=2E =20 If you have staff with vacuum experience you can probably fix this=20 yourself=2E Help is also available from Denton by contacting Jim Felc= o=20 at 609-439-9100=2E He may also be able to suggest a repair person in=20 your area=2E =20 Good Luck =20 =20 John Humenansky Braun Intertec Microscopy Department 6875 Washington Ave=2E So Minneapolis, MN 55439 612-942-4822
Mr-Received: by mta RANDD; Relayed; Wed, 27 Aug 1997 08:05:50 -0500 (CDT) Mr-Received: by mta MCM$RAND; Relayed; Wed, 27 Aug 1997 08:05:52 -0500 (CDT) Mr-Received: by mta RANDD; Relayed; Wed, 27 Aug 1997 08:05:55 -0500 (CDT) Alternate-Recipient: prohibited Disclose-Recipients: prohibited Content-Return: prohibited
Damian,
We have a Philips XL30 FEG w/ EDS and we get plenty of current (I have measured over 35 nA) that is very stable over time. In addition, we routinely image at 1-3 kV. All around it is a versatile instrument capable of excellent performance. I would say an FESEM is very capable of doing EDS.
On your comment about catalysts and accelerators, the JB-4 kit calls the benzol peroxide powder a catalyst and it is added to the infiltration solutions as well as the final solution. But, in the final mix, it is mixed in just prior to the solution B. JB-4 polymerization is done either at room temp or at 4 degrees centegrade. As a catalyst often accelerates a reaction, is there really a difference in calling it an accelerator or a catalyst? Just wondering.
Karen Pawlowski UT Dallas, UT Southwestern Medical Center
On Tue, 26 Aug 1997, IAN HALLETT wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I have run into a couple of occassions of premature polumerization } in the 10 or more years we have used LR White resin. In both cases } the resin was old, ie over 1 year but was not associated with } osmication. An possible explanation was suggested to me by Roy } Gillett of London Resin a number of years ago who definitely } recomended a shelf life of 12 months for catalysed resin. } } Quote } "The reason this pre-polymerisation occurs only with tissue must be } something to do with a tissu constituent catalysing polymerisation. } Older resin is much more susceptibe to this that fresh monomer becaue } of the significant polymer growth that will inevitably have occurred } in the monomer. The most likely 'endogenousd catalyst' from } previous experience is likely to be an amine or peroxide moiety in } the tissue" } } We have had no problems since switching to buying uncatalysed resin } and making up a new bottle as we run out of the old. } } One point I have noticed in the discussions to date is some } ambiguity between calalyst and accelerator. From my understanding } the catalyst (benzoyl peroxide powder) must be added 24 hours before } you start using a batch of resin and is necessary for both thermal } (oven) and "cold" polymerization. The accelerator on the other hand } is added to the final resin change for rapid "cold" polymerization } without using an oven. } } Ian } } } } Ian Hallett } HortResearch } Mt Albert Research Centre } Private Bag 92 169 } Auckland, New Zealand } Fax 64-9-815 4201 } Telephone 64-9-849 3660 } EMail ihallett-at-hort.cri.nz }
This question has been around several times, so you might want to check the log of past postings.
We have run our facility as a fee-for-service lab for over 15 years now. Several things to keep in mind (debated hotly previously- so I hope I am not reopening the debate) is not to use University Subsidy to undercut your price to users outside your university.
Now, as to how do we determine fees for microtomes, etc. We know our service contract costs, we add depreciation to this and divide the total by the number of useable hours the instrument is available for use. This establishes our break-even cost. We add about 10% to this, since the instrument is down at times for service, etc.
Every piece of equipment has a sign-in/sign-out sheet. We send bills out once a month.
Consumables are charged at an average cost for a procedure. I.e. if you do a negative stain we know about what it costs for grids, stains, etc.
When someone in the laboratory does the work this cost is added to the charge.
Importantly, my consulting expertise comes free to University users. However, my time is charged for any outside work we do.
I would again urge you to consult past posts with regard to what is legal, ethical, and kind with regard to competition with outside suppliers of EM service who do not have the benefit of a University behind them. There are some fervently held beliefs on this score which should be heeded.
I hope this helps-
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-bgsm.edu * **************************************************************
On Wed, 27 Aug 1997, Richard Lander wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear all, Message on behalf of Allan Mitchell; } } Subject; Charging for usage in the multiuser EM Unit } } We are a university based multi-user EM Unit that is moving towards the } 'real' world. Presently, all university users of our EM Unit are charged } for the consumables they use (based on an honesty system) but charged for } nothing else. Our bean counters are now requiring us to introduce } equipment usage charges, labour charges and occupancy charges. At this } point I have no idea how much they expect us to recover, the wording that } is used is that those who use the service should contribute to the cost of } running the service. Contribute is the key word. } } I have no problem with this concept at all. My problem is however, how do } we record the many 'chargables' and I am hoping that those of you out there } who have introduced such charging systems to your Units and those who are } working with such systems can help me. } } Our EM Unit is multi-user Unit where 80% of the users come to the Unit and } do the work themselves after we have trained them. } } Equipment Charges } Charging for electron microscope usage is simple enough taken just from a } HT counter however how do you charge for the ultramicrotomes, microwave } oven, tissue processor, pH meters, osmometer, enelargers, photo printer etc } without having a lot of clipboards all over the place ? } } Occupancy } How do you keep records of all the users coming and goings from the Unit so } that a sensible occupancy charge can be made ? } } Labour Charges } Easy to do by introducing a 'Job Card' system for the work we do. } } All help and suggestions appreciated } } Regards } } Allan } } } } ----------------------------------------------------------------------- } Richard Lander } Electron Microscope Technician } South Campus Electron Microscope Unit } Otago School of Medical Sciences } P.O. Box 913 } Dunedin } New Zealand. } Tel. National 03 479 7301 Fax. National 03 479 7254 } } "Southernmost EM Unit in the World!" } ------------------------------------------------------------------------ } } }
In message {v02130500b028a30961ec-at-[128.32.175.193]} Paula Sicurello writes: } } I have a Denton 502A vacuum evaporator that doesn't suck like it } used to. I've tried looking at it and I'm getting pretty frustrated. The } problem is it doesn't ROUGH Pump like it used to, it acts like it has a } leak.
If it dopesn't rough pump, you probably DO have a leak or mechanical pump has a problem and it must be rather major and so perhaps easy to find. I have not maintained a Denton, but here are a few general things you can check, in case you havn't thought if these already.
1. Figure out ways to isolate parts of the system to check for leaks. To check if the leak is in the jar and its seal, or below the stage in the guts of the system, put a plug (eg. O-ringed fitted plate, like you have in your service kit for SEM or TEM) over the opening in the stage baseplate to the pumping system below and see if you can pump down.
2. If you can pump down, then the leak must be above the baseplate. If the bell jar has a rubber gasket mounted along the bottom edge, pull it back to check for chipping of the glass edge, or a crack. This can happen due to putting the jar onto the baseplate too hard and hitting a metal fitting on the stage. If so, clean out glass fragments, clean damaged edge with ethanol or acetone, dry with canned gas, fill with silicone bathtub caulk and gently reposition rubber seal. Then pump down just a little so that seal sets up under mild vacuum.
3. If system doesn't pump down as result of test in #1 above, then you have many more places to look!! : {( I suggest you check your mecnanical pump first. Can you pump down in the foreline area with the mechanical pump isolated from the rest of the system? If not, is the pump's oil level too low? Or if you havn't changed the mechanical pump's oil for a long time, try that for starters, as over time you can get moisture building up in the oil and that will certainly erode pump performance, usually at high vacuum end, tho. Do two changes, the first as a quick rinse - run on first oil change for about 5 minutes, then change it a second time.
4. Check all rubber or plastic hoses. First look at areas where tubing is clamped onto metal fittings, as hoses can eventually split there. Or tighten up hose clamps a little.
5. Valves and air inlets can also malfunction due to wear over years of use, or the lubrication drying out. You may need to open them up, clean and lubricate them.
Hope this helps and good luck.
--
Gib Ahlstrand, MMS Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
A few years back, I contacted Denton about service and they had someone on the west coast who could do it. I believe it was Jim Falco, but you can contact Denton to find out for sure. At that time, I talked to Rob Specht (Marketing Manager) at (609)424-1012. Hope this helps.
Regards,
Bob *************************** Bob Citron Chiron Vision Claremont, CA USA (909)399-1311 Bob_Citron-at-cc.chiron.com ***************************
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello All!
I have a Denton 502A vacuum evaporator that doesn't suck like it used to. I've tried looking at it and I'm getting pretty frustrated. The problem is it doesn't ROUGH Pump like it used to, it acts like it has a leak. Does anyone out there know of a company or individual who does service calls on Denton Vacuum evaporators that's on the west coast, preferrably in the San Francisco Bay Area? Please help me! I'm close to just hitting the thing with a hammer! Thanks for any information you might give me.
Frustrated in Berkeley,
Paula = ) psic-at-uclink4.berkeley.edu
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
I've gotten a couple of private emails on this. First, I didn't intend to imply that FE guns are necessarily less stable than LaB6 or W; this is not the case for many newer instruments, as several later postings on this topic have pointed out.
Second, a few people asked how you would correct an element map for current drift, regardless of gun type. One way is to collect a map of current per pixel corresponding to the element maps, by running a picoammeter signal through a voltage-to-frequency (V2F) converter. Most EDX systems have inputs for an external rate signal of this type. Scaling the X-ray counts using the current map counts at each pixel does the trick visually, although you're still left with pixel-to-pixel precision differences if you want to quantify the maps. Normalization can't fix that.
take a deep breath, relax, put down the hammer, pick up the phone and call Jim Falco, a service guru at Denton (609)439-9100. When I had a high vacuum leak in my unit, he walked me through step by step troubleshooting over the phone and I was able to find the problem (a microcrack in the flange in the main valve unit). He told me how much for the repair part and step by step how to install it, all of which worked out as he predicted.
I was very pleased with the assistance I received from Denton (I don't have any kind of service agreement or personal financial interest with them) and hope this type of after sales assistance is standard company policy that will continue in the future.
steve
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dr. Steven Barlow, Associate Director EM Facility/Biology Department 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/EM_Facility
--IMA.Boundary.530596278 Content-Type: text/plain; charset=US-ASCII Content-Transfer-Encoding: 7bit Content-Description: cc:Mail note part
Keith,
I too have used HMDS very successfully for a large number of bacteria species. HOWEVER, some bacteria require different protocols than others. Some species came through very well preserved with no evidence of shrinkage, others looked like they had been air dried from water! But with a little experimenting, you should find the right times. I used ethyl alcohol (I would think that acetone should be good as well): 25%, 50%, 70%, 80%, 95%, 100% x 3 15 min each, then a 3:1 EtOH:HMDS, 1:1, and 1:3 15 min ea, then HMDS 3X 15 min each, drain, then place in desicator to dry. Drying at 60deg ok too.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Keith,
First, do you *have* to CPD? I've used HMDS very successfully for bacteria, in some cases retaining the "slime" sheath (that depends more on dehydration). 5 min. changes, 100% Ac =} 1:1 Ac:HMDS =} 3 X 100% HMDS, dry at room temp or 60 C
Air drying straight from acetone can work for bacteria nicely--do you have time to experiment?
For CPD, use 3 to 5 changes of CO2, with 2 to 5 minutes soaking in CO2 between changes (time by slime coat). Your real problem is going to be turbulence on filling and emptying the chamber, so this will have to be done *very slowly* so has not to disturb the coverslips.
Note: this is Ed Basgal's design and idea!: A cover-slip holder can be quickly made by cutting notches in a polyethylene or glass tube (not tygon), just wider that the thickness of the coverslips, so the slips fit down into the notch:
_ |_ {--cover slip fitting into notch | | | |__|
place tube into a polyethylene syringe body or centrifuge tube that's been fenestrated by a mad perforator. Cap both ends.
Phil
} I have to dry some bacteria grown on glass coverslips shortly for SEM } from acetone to liquid CO2 i.e. critical point drying. I would appreciate } advice in the next 24 hours about what times are recommended in the } drier. } } Unfortunately, the 'customer' has used quite a few 22 x 22 mm } coverslips and I only have the original Polaron drier, without any } specially-made coverslip holders, so it looks like three will fit in gauze } baskets which I use for baby squid. } } Regards - Keith Ryan } Plymouth Marine Lab., UK
(IMA Internet Exchange 2.1 Enterprise) id 00194947; Tue, 26 Aug 97 19:56:17 -0500 Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by ns2.baxter.com (8.8.0/8.8.0) with SMTP id UAA24598; Tue, 26 Aug 1997 20:03:38 -0500 (CDT) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id RAA08994 for dist-Microscopy; Tue, 26 Aug 1997 17:16:31 -0500 Received: from ux1.cso.uiuc.edu (ux1.cso.uiuc.edu [128.174.5.59]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id RAA08991 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 26 Aug 1997 17:16:30 -0500 Received: from [130.126.25.7] (florence-8.slip.uiuc.edu [130.126.26.132]) by ux1.cso.uiuc.edu (8.8.5/8.8.5) with SMTP id RAA07717; Tue, 26 Aug 1997 17:23:45 -0500 (CDT) X-Sender: oshel-at-staff.uiuc.edu Message-Id: {v02120d01b0290a6b20d7-at-[130.126.25.7]} Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
On Wed, 27 Aug 1997, Linda Fox wrote:
} Date: Wed, 27 Aug 1997 07:33:38 -0500 } From: Linda Fox {lfox1-at-wpo.it.luc.edu} } To: Microscopy-at-sparc5.microscopy.com } Subject: enhancing herpes } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello again, } I have rec'd. a sample for TEM that may be infected by a herpes } virus. I am wondering if there is a way of enhancing the tissue with } any stains (tannic acid etc.) to optimally show the virus. The tissue } has been fixed in 4%GA, cacodylate buffer. I will be processing } today, so I appreciate any info asap. Sorry for the short notice. } Thanks } Linda Fox lfox1-at-wpo.it.luc.edu
Unless you're going to do immunoEM, any fixes using glut, Os, and UA will be fine. Infected cells should have 100 nm nucleocapsids in the nucleus and 200 +/- nm complete virions in the cytoplasm. Nucleocapsids may bud from nuclear, intracytoplasmic, and plasma membranes. You can cut thick sectins and stain them with toluidine blue, Paragon, multiple stain, etc. and look for abnormal areas before selecting areas for thins. Contact me directly if you have questions.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
I remember there being some mention in the past of techniques to bring a computer display into harmony with the a printout regarding color hues, intensity, etc. Does anyone remember this or have any suggestions to add? I seem to recall there being certain software tricks or packages involved.
For background, we just purchased an Epson Photo Stylus color printer and a Polaroid Sprintscan 35 with PathScan Enabler, which seem to do a great job for scanning and printing light microscope slides to get an overview of the whole section. The only problem is that the colors on the printout don't exactly match the screen display (computer running Windows 95), and neither are perfect in comparison to seeing the slide under the microscope. We could live with what we get on the screen if only the printouts would match. A colleague has been playing with settings in Adobe Photoshop (CMYK vs. RGB) and with printer ICM (??) setting, but they only seem to make things worse.
Oh and I should mention we haven't yet contacted the vendors to get their input so perhaps they will have a simple suggestion. But in the mean time any help from the list would be appreciated!
Karen
-- Karen Zaruba kszaruba-at-mmm.com 3M Company, 3M Center Bldg. 270-1S-01 St. Paul, MN 55144 "The opinions stated above are my own, not necessarily 3M's"
I am looking for a US supplier for Nuclepore gridded filters. I would also like the phone number or web site for the Nuclepore Corporation. Please send the information to my email address: gondo-at-sprynet.com.
Analytical Consumer, a monthly newsletter for analytical laboratories, is doing a survey of users of all types of scanning probe microscopes. Each month, we survey users of one kind of analytical equipment, asking what they use, why they bought it, and what they think of its performance and support. Then we report what the labs say. We are not associated with any organization or instrument company, and we accept no advertising.
If you use SPM and would be willing to be part of the survey, send me an e-mail (address below), and I'll send you the short list of questions. Every one who replies receives a copy of the final report, of course.
Thanks, Jo Rita
Jo Rita Jordan, PhD Editor and Publisher Analytical Consumer
There were some questions a while back on the list about the file format used by the Oxford/Link ISIS .
I'm currently using the export to EMMFF to get the data out of the link system, but it would still be useful to have info on the format for batch conversions of large data sets. ( Or for when you don't have access to the ISIS software. )
If anyone has any more authoritive info, I'ld like to hear it. In the meantime, from reverse engineering some of our data files and comparing them with the EMMFF output, I have: ( Note: I don't know what variable info may be in the header that might be different in a different setup -- this is from a small sample of files. )
Partial Format of .SPE files: ( offsets are zero based )
Spectrum is the last 1024 ints of the file.
CHOFFSET is float at word [384] RealTime is int at word [130] LiveTime is int at word [128] ( or are these perhaps the low order of a double-word long int ? ) RealTime and LiveTime are both in microsecs.
A string containing title and analyst starts at byte[10] ( don't know if this is fixed or variable length, and whether it's split into two separate fields. )
This may be enough for a partial conversion. If anyone has any more info to add, please let me know.
[ BTW: I've found Python {http://www.python.org/} to be a good tool for this sort of reverse-engineering/investigative-programming. The array classes even have byteswap() methods, which, since I'm doing this conversion on a Mac, and ISIS files come from a PC, come in quite handy. I also have a new version of a DTSA -} EMMFF batch converted written in Python, if anyone is interested. ]
---| Steven D. Majewski (804-982-0831) {sdm7g-at-Virginia.EDU} |--- ---| Department of Molecular Physiology and Biological Physics |--- ---| University of Virginia Health Sciences Center |--- ---| P.O. Box 10011 Charlottesville, VA 22906-0011 |--- All power corrupts and obsolete power corrupts obsoletely." - Ted Nelson
} Date: Wed, 27 Aug 1997 09:13:28 -0500 (CDT) } From: {kna101-at-utdallas.edu} } To: IAN HALLETT {ihallett-at-hort.cri.nz} } Cc: microscopy-at-sparc5.microscopy.com } Subject: RE: LR White Question- Premature Polymerization.
} Ian, } } On your comment about catalysts and accelerators, the JB-4 kit } calls the benzol peroxide powder a catalyst and it is added to the } infiltration solutions as well as the final solution. But, in the final } mix, it is mixed in just prior to the solution B. JB-4 polymerization is } done either at room temp or at 4 degrees centegrade. As a catalyst often } accelerates a reaction, is there really a difference in calling it an } accelerator or a catalyst? Just wondering. } } Karen Pawlowski } UT Dallas, UT Southwestern Medical Center } Karen
In the case of LR White resin there is a difference in that the catalyst powder must be added to the resin well before it is used (at least 24h) and is used to "activate" the bottle of resin to allow polymerisation. The shelf life of the resin starts from this time. Initially all LR White resin was sold catalysed which caused some problems with old stock.
The accelerator is added only when a "cold" cure is requried - I'm not sure what it is (comes as a liquid), we never use it. The "cold" cure is a strong exothermic reaction hence the "" around cold.
Ian
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-849 3660 EMail ihallett-at-hort.cri.nz
John J. Bozzola wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Anyone know of a review article or bibliography on cathodoluminescence? I } have one dating back to 1977 but I imagine there must be a more recent one. } I did a lit search but came up negative. Any help would be appreciated. } Thanks. } } #################################################################### } John J. Bozzola, Ph.D., Director } Center for Electron Microscopy } Neckers Building, Room 146 - B Wing } Southern Illinois University } Carbondale, IL 62901-4402 } U.S.A. } Phone: 618-453-3730 } Fax: 618-453-2665 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/departments/shops/cem.html } ####################################################################
A quick follow-up to John's question re: cathodluminescence bibio..
There was a device, sold in the UK, which did cathodluminescence on a light microscope. Does anyone have lit, info, source of supply for US?
At 01:05 PM 8/26/97 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The disadvantages are: With cold field emission the current fluctuations from the gun have to be compensated when you are doing quantitative EDS
We find we dont do any quant with our 4500 (we also have a Cameca Probe) so that is no concern
Cold field has the lowest beam current of any emitter. But it is still enough to saturate the EDS detector and also for our cathodoluminescence system
Information to the contrary from sellers of Hot field emitter guns is too pessimistic
} 3. Would we be better off using tungsten filament source?, LAB6 } source? for EDXS? } In practice, No.
} 4. Imaging is also very important. As a reference point, we have a } JEOL 6300F and are very pleased with the imaging at low keV. Are the } current tungsten or LAB6 source microscopes being sold capable of } providing the same level of image quality as a FE source microscope in } the 1-5 keV range? }
Absolutely NOT!. We had a S-900 FESEM and bought the 4500 so we could apply the brilliantly improved resolution and low kV we were getting to large specimens. It as forever changed how we do our microscopy.
Mel Dickson President, Australian Society for Electron Microscopy Director, Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
Those who have tried to reach the Web site for next year's International Congress on Electron Microscopy to be held in Cancun Mexico may have been disappointed in the last couple of days. I have just called Mexico. The Web site is being updated. It is expected to up again early next week at
http://icem.inin.mx
Plan on attending the meeting! .
** Alwyn Eades Center for Microanalysis of Materials University of Illinois at Urbana-Champaign Phone 217 333 8396 Fax 217 244 2278 eades-at-uimrl7.mrl.uiuc.edu (NB those are letter l not ones) **
I understand that information on the next International Stereology Course was posted earlier on the listserver. I have just subscribed and would be very grateful if someone could forward me the posting or any relevant information.
Many Thanks Teeba Lundy lundyt-at-agresearch.cri.nz Wallaceville Animal Research Centre Upper Hutt NEW ZEALAND
anyone out there who has a philips 400 tem that could give me simple instructions for alignment and filament saturation? i'm new in a lab that has no one (tech retired and just left) that is familiar with this instrument. i've been doing okay so far but it would be nice to hear from someone who has this instrument.
It is quite impossible to charge for each grid, piece of glass etc. For many years I used a scheme to just charge for EM time, sheet film and prints. The prints were charged by weighing customer weighing the current paper box before and after and recording this nearest gram in a logbook. Weighing the box doubled the returns on the previous "I used so many sheets" - people are honest when you can verify their honesty. Paper paid for developer and fixer and about 15% extra covering 35mm TEM film too. SEM 120 roll film was charged for, but a 35mm camera with 50 ASA film and a half decent macro lens is just as good on the SEM and can be fully automatically integrated.
The SEM/ TEM charges had to pay for all other consumable in the lab. Filament time not kv should be used because for most situations, especially high res its better to leave the kV on. For the probe its best to charge for time booked, because the filament too needs to remain on.
Its far more efficient and cost effective to purchase centrally. If every user needs to buy grids, osmium and buffer etc. an awful waste occurs (which should be good for my business). Try to teach users to use small volumes. One problem are lab users who just prepare their samples in your lab and look at them elsewhere - or just use LM. Overall this system worked well for many years.
I charged $12 for EM hour and received a maintenance grant of $12000 which was cut to $5000 after some years and then a new, half-baked VC (CEO/ or president) declared: Its user pays; no maintenance. That is when the arguments get interesting. Note a poor CEO makes up policy and is not interested to make policy work in a real world.
That lab had a considerable teaching function for undergrads, but also had a hundred grad students a year. If they spent 20% of their time in the EMU on related activities, the Unit should have been top ranking in box top counts. The 3 EM busy unit was run with one and a half staff only. That is a bit of a tricky thing to handle. As a result, in terms of user-pays would have excelled 'in the real world'. Admin, library and computing took half of all Gov fees, about$20000 annually for the students. The departments received the other half and the departments returned generally $1000 to PhD students.
Do the arithmetic any way and it would be obvious that the EMU more than paid its way. What that VC wanted was to count every $ twice. User pays was a scheme to get more money from nothing, which is a poor economic principle.
The EM Committee voted against increasing hourly fees. To absorb the lost maintenance would have meant at least a doubling of hourly fees. Higher fees result in less usage. Eventually the VC wanted to charge for labour too. That would have required charges of about $70/filament hour.
Get me right, I am not against 'user pays'. It could have worked if the students received say $5000 of there funding to spent on campus based research. In my experience the phrase 'User pays' is used to give legitimacy to fancy schemes, like charging effectively twice, increasing resources for admin. and decreasing the dollar going to the universities primary functions: Learning and research.
That VC, after twelve long years was shown the door - but he still received a 'golden hand shake'. His legacy is a smallish uni with a debt of $29 million and a lot of run down facilities and a very demoralised staff.
An administrations role is to facilitate learning and research. Sure, the place has too operate within budget, but beware of anything that makes administration more complex. Charging and other admin tasks may be necessary, but the effort detracts from research and to an extend is counter-productive. Charging must make a lab less efficient, so use the simplest system that can be devised.
Beware of administrators spouting half-smart slogans. Charging is meant to extract more money from somewhere, but nobody has put more money into the system. So it is just a way of re-allocating resources, usually do the detriment of the university's primary functions. Cheers, its no good crying about that. Jim Darley
This is really rather out of topic, but would any of you by any chance know of an existing listserver (just like Microscopy) that discusses ischemia, reperfusion injury, animal and cellular models and related topics?
Hope you could help.
Thanks!
Ginny
------------------ GINNY E. CRUZ Section of Immunopathogenesis, Institute of Immunological Science Hokkaido University : Kita Ku Kita-15 Nishi-7 Sapporo City 001 JAPAN Tel. No.: (011)716-2111 Ext. 5120 Fax No.: (011)736-9836 e-mail: gcruz-at-imm.hokudai.ac.jp ------------------
} Dear All of the Microscopy Net: } } This is really rather out of topic, but would any of you } by any chance know of an existing listserver (just like } Microscopy) that discusses ischemia, reperfusion injury, } animal and cellular models and related topics? } } Hope you could help. } } Thanks! } } Ginny } } } ------------------ } GINNY E. CRUZ } Section of Immunopathogenesis, Institute of Immunological Science } Hokkaido University : Kita Ku Kita-15 Nishi-7 Sapporo City 001 JAPAN } Tel. No.: (011)716-2111 Ext. 5120 Fax No.: (011)736-9836 } e-mail: gcruz-at-imm.hokudai.ac.jp } ------------------
------------------ GINNY E. CRUZ Section of Immunopathogenesis, Institute of Immunological Science Hokkaido University : Kita Ku Kita-15 Nishi-7 Sapporo City 001 JAPAN Tel. No.: (011)716-2111 Ext. 5120 Fax No.: (011)736-9836 e-mail: gcruz-at-imm.hokudai.ac.jp ------------------
Hello all ! I have a problem in determining the divergence angle of our TEM Jeol 2010=20 for the different modes of this microscope. Indeed, three divergence angles= =20 are available for imaging studies called : alpha=3D1, 2, 3, which should be= =20 used from high magnification values (} mag 400kX) to low maginifcations=20 ( {100 kX) and then should correspond to increasing divergence angle from 1= =20 to 3. I have tried to measure this angle, by classical method (on E.D=20 patterns obtained with a condensed electron beam), but obtained values were= =20 absurd, in total contradiction with expected ones. Jeol company from France= =20 cannot give me any explanation for this, nor another measurement procedure.= =20 Can anyone help me ? Thanks Bernadette Domenges CRISMAT-ISMRA Boulevard du Marechal Juin 14050 CAEN CEDEX - FRANCE E-mail : domenges-at-crismat.ismra.fr Fax : (33) 02 31 95 16 00 T=E9l : (33) 02 31 45 26 32
DuPont Medical Products Biotechnology Systems Division 31 Pecks Lane Newtown, CT 06470-5509
Tel: (800) 551-2121 Fax: (203) 270-2166
http://www.sorvall.com
They don't distribute through other suppliers or distributors.
Regards,
Rick Powell
****************************************************************** * PLEASE NOTE MY NEW E-MAIL ADDRESS: rpowell-at-mail.lihti.org * * NANOPROBES GENERAL E-MAIL ADDRESS: nano-at-mail.lihti.org * * * * NANOPROBES, Incorporated | Tel: (516) 444-8815 * * 25 East Loop Road, Suite 124 | Fax: (516) 444-8816 * * Stony Brook, NY 11790-3350, USA | nano-at-mail.lihti.org * * * * NOW EASY TO FIND ON THE WEB: http://www.nanoprobes.com * ******************************************************************
} Hi All, } } I need to find a supplier for the Sorvall GLC-2, general } laboratory centrifuge. We inherited one and need some more test tube } holders. Thanks. } } Karen Pawlowski } UT Dallas/UTSW Medical Center
} } } } Dear All of the Microscopy Net: } } } } This is really rather out of topic, but would any of you } } by any chance know of an existing listserver (just like } } Microscopy) that discusses ischemia, reperfusion injury, } } animal and cellular models and related topics? } } } } Hope you could help. } } } } Thanks! } } } } Ginny } } } } } } ------------------ } } GINNY E. CRUZ } } Section of Immunopathogenesis, Institute of Immunological Science } } Hokkaido University : Kita Ku Kita-15 Nishi-7 Sapporo City 001 JAPAN } } Tel. No.: (011)716-2111 Ext. 5120 Fax No.: (011)736-9836 } } e-mail: gcruz-at-imm.hokudai.ac.jp } } ------------------ } } ------------------ } GINNY E. CRUZ } Section of Immunopathogenesis, Institute of Immunological Science } Hokkaido University : Kita Ku Kita-15 Nishi-7 Sapporo City 001 JAPAN } Tel. No.: (011)716-2111 Ext. 5120 Fax No.: (011)736-9836 } e-mail: gcruz-at-imm.hokudai.ac.jp } ------------------
------------------ GINNY E. CRUZ Section of Immunopathogenesis, Institute of Immunological Science Hokkaido University : Kita Ku Kita-15 Nishi-7 Sapporo City 001 JAPAN Tel. No.: (011)716-2111 Ext. 5120 Fax No.: (011)736-9836 e-mail: gcruz-at-imm.hokudai.ac.jp ------------------
Some people mentioned that a premature polymerisation of the resin might be avoided if the catalyst is added to the resin just prior to the embedding.
I looked into several catalogues from different vendors but only found kits where the resin (presumably containing the catalyst) and the accelerator were offered.
I would like to ask for inputs which company sells LR White with the catalyst seperate (preferably in Europe).
Hans-Martin ************************************************************** Hans-Martin Vaihinger Ruhr-University of Bochum Comparative Endocrinology Research Section Building ND 5/37 44780 Bochum GERMANY ********************************************************* phone ++49 234 700 4329 fax ++49 234 709 4551 e-mail Hans-Martin.Vaihinger-at-Ruhr-Uni-Bochum.de
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello-
I am looking for a US supplier for Nuclepore gridded filters. I would also like the phone number or web site for the Nuclepore Corporation. Please send the information to my email address: gondo-at-sprynet.com.
I buy the LR White from Ted Pella Inc. and it comes separate.
Bob
On Thu, 28 Aug 1997 Hans-Martin.Vaihinger-at-Ruhr-Uni-Bochum.de wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } To the people discussing LR White problems } } Some people mentioned that a premature polymerisation of the resin } might be avoided if the catalyst is added to the resin just prior to } the embedding. } } I looked into several catalogues from different vendors but only } found kits where the resin (presumably containing the catalyst) and } the accelerator were offered. } } I would like to ask for inputs which company sells LR White with the } catalyst seperate (preferably in Europe). } } Hans-Martin } ************************************************************** } Hans-Martin Vaihinger } Ruhr-University of Bochum } Comparative Endocrinology Research Section } Building ND 5/37 } 44780 Bochum } GERMANY } ********************************************************* } phone ++49 234 700 4329 } fax ++49 234 709 4551 } e-mail Hans-Martin.Vaihinger-at-Ruhr-Uni-Bochum.de }
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am trying to find Roger Waldock. I recall that he sold EDM (spark machining) instruments for slicing and coring of samples for metallography, TEM,...etc. sample preparation. The instrument that we are interested in buying is the Servomet. I believe that it is/was produced in Cambridge, England. If anyone can help in locating Roger and or who sells these in the USA I would appreciate it.
Thanks, Mark A. Wall L-350 Chem. & Mat. Sci. Dept. Lawrence Livermore National Lab 7000 East Ave. Livermore, CA 94550 USA ph. 510 423-7162 fx. 510 422-6892
} The instrument that we are interested in buying is the } Servomet. I believe that it is/was produced in Cambridge, England. If anyone } can help in locating Roger and or who sells these in the USA I would } appreciate it. } I do, however, have some copies (circa 1990) of EDM Digest, more recent issues of American Tool, Die & Stamping News and Metal Forming. I'd be happy to send you any or all of these if you could use them. Yours, Bill Tivol
My SEM books are quite vague regarding what channeling contrast on the SEM is. Can anyone out there tell me 1) What is the mechanism of channeling contrast on the SEM? 2) What detectors/equipment is necessary to do channeling contrast on the SEM? 3) What are the sample and sample prep. requirements for channeling contrast on the SEM?
DuPont Medical Products Biotechnology Systems Division 31 Pecks Lane Newtown, CT 06470-5509
Tel: (800) 551-2121 Fax: (203) 270-2166
http://www.sorvall.com
They don't distribute through other suppliers or distributors.
Regards,
Rick Powell
****************************************************************** * PLEASE NOTE MY NEW E-MAIL ADDRESS: rpowell-at-mail.lihti.org * * NANOPROBES GENERAL E-MAIL ADDRESS: nano-at-mail.lihti.org * * * * NANOPROBES, Incorporated | Tel: (516) 444-8815 * * 25 East Loop Road, Suite 124 | Fax: (516) 444-8816 * * Stony Brook, NY 11790-3350, USA | nano-at-mail.lihti.org * * * * NOW EASY TO FIND ON THE WEB: http://www.nanoprobes.com * ******************************************************************
} Hi All, } } I need to find a supplier for the Sorvall GLC-2, general } laboratory centrifuge. We inherited one and need some more test tube } holders. Thanks. } } Karen Pawlowski } UT Dallas/UTSW Medical Center
I can't help you much with locating Roger, but I thought I would make you=
aware that we do offer a small Spark Erosion Unit for TEM applications. = It is an inexpensive unit and is designed for small samples. You can get so= me limited information on our web site or I would be pleased to send you a complete data sheet on it.
David Henriks TEL: 800-728-2233 (toll-free in USA) South Bay Technology, Inc. 714-492-2600 1120 Via Callejon FAX: 714-492-1499 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 "Mark Wall" } I am trying to find Roger Waldock. I recall that he sold EDM (spark machining) instruments for slicing and coring of samples for metallography, TEM,...etc. sample preparation. The instrument that we are interested in buying is th= e Servomet. I believe that it is/was produced in Cambridge, England. If anyone can help in locating Roger and or who sells these in the USA I would appreciate it.
Every now and again, with semi-thin sections made from Epon-araldite plastic (0.5 microns thick) of nerve or muscle, I get a wrinkled result that looks as though the entire section has fractured. I've tried adjusting the temperature of the hot plate, or even giving it only 30 seconds on the hot plate, I've tried cutting wide rather than long, I've tried trimming away all the excess plastic around the tissue area, and I've tried cutting thinner sections, but in all cases, once I get this "cracked" appearance to the section, I have great difficulty cutting it to give me secitions that don't have these networked wrinkles.
Does anyone have any thoughts as to how I might avoid this problem, before I go insane!!
Garry, It has been suggested to me to try using 40% acetone/dH2O instead of dH2O when semi-thin sectioning. I tried it with just Spurr's and it worked pretty good (but this was a botanical sample). Good Luck! Tracey Pepper Iowa State Univ.
Hans-Martin.Vaihinger-at-Ruhr-Uni-Bochum.de wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } To the people discussing LR White problems } } Some people mentioned that a premature polymerisation of the resin } might be avoided if the catalyst is added to the resin just prior to } the embedding. } } I looked into several catalogues from different vendors but only } found kits where the resin (presumably containing the catalyst) and } the accelerator were offered. } } I would like to ask for inputs which company sells LR White with the } catalyst seperate (preferably in Europe). } } Hans-Martin } ************************************************************** } Hans-Martin Vaihinger } Ruhr-University of Bochum } Comparative Endocrinology Research Section } Building ND 5/37 } 44780 Bochum } GERMANY } ********************************************************* } phone ++49 234 700 4329 } fax ++49 234 709 4551 } e-mail Hans-Martin.Vaihinger-at-Ruhr-Uni-Bochum.de
We are on such company and we sell direct to Germany or we can give you the names of one of our agents in Europe if you would like to buy from them. Please let me know.
Currently we have a JEOL JAMP 30 auger microprobe with Link AN 10/55S light element detector for anyone who is interested. Please email me at this address and indicate "microprobe" in your subject header or fax an information request to M.W. Rigler at MAS, Inc. 770-368-8256. The fax will ge a faster response.
Cheri Moss asked for a reference on channeling contrast in SEM. I suggest trying Chapter 3 in the book 'Advanced Scanning Electron Microscopy & X-Ray Microanalysis', by Dale Newbury, et. al., Plenum Press, 1986,ISBN 0-306-42140-2.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
Regarding the necessity of charging for EM services, it is now officially near the end of the silly season, so I thought I would like to share with you all this little story. I hope you find it amusing!
I will go back a few hundred years to the Middle East, to visit a man called Nasr-ed-Din. He was a local official who worked as a town clerk, magistrate, and letter writer, since he was the only learned man in his town or village. A man in his position would have been called a Mullah in Persian, and in Turkish a HODJA, which is how I will call him from now on.
The Hodja had heard about some Sufis who claimed that they could achieve enlightenment by going for a long time without food, and he thought that this sounded like a good idea. But it also seemed a bit dangerous, so he thought he would try it on his donkey first. He normally gave the animal 20 scoops of grain a day, so for the next week he gave it 19, then the week after 18, and so on. By the time he had reached 10, the donkey was beginning to get weak and to wander about as if in a dream.
"Excellent" thought the Hodja, "it is beginning to learn the art of meditation!"
But just before he was getting down to 3 scoops a day, the donkey died.
"Inconsiderate beast" he cried. "Dropping dead before the experiment was complete. If only it had lived a few weeks longer, it might have achieved full enlightenment!"
All over the world, in university science departments, numbers have been falling. Typically, since around 1970;
- Student numbers have dropped to about 75% of their original number;
- Teaching staff to about 50%;
- Technical staff to about 25%.
This last figure has put an enormous burden on the research as well as the teaching effort. Whenever A retires, the government body says "You don't need a replacement - can't B and C share his job? And so on ... it is predicted that by the year 2010 such a department will be running on no staff at all! But it must expire before then, once it gets below a critical (m)ass.
+------------------------------------------------------------------------+ | 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 | +------------------------------------------------------------------------+
We handle a fair number of muscle, and nerve biopsies and are usually able to avoid wrinkles by flattening with a heat pen while the section is still floating in the boat. Although, every so often we do have a specimen that has such internal elasticity (tension) that occasional wrinkles do occur. These are not apparent in the ultra-thin section (also flattened with a heat pen). You may want to check the specimen protocols if this is a consistent problem. Ensure that the specimen is clamped or tied with the correct amount of tension when accepted from surgery and during fixation.
Peter O. Steele, PhD All Children's Hospital St. Petersburg, FL
} } } Garry Burgess {GBurgess-at-exchange.hsc.mb.ca} 08/28/97 02:25pm } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Every now and again, with semi-thin sections made from Epon-araldite plastic (0.5 microns thick) of nerve or muscle, I get a wrinkled result that looks as though the entire section has fractured. I've tried adjusting the temperature of the hot plate, or even giving it only 30 seconds on the hot plate, I've tried cutting wide rather than long, I've tried trimming away all the excess plastic around the tissue area, and I've tried cutting thinner sections, but in all cases, once I get this "cracked" appearance to the section, I have great difficulty cutting it to give me secitions that don't have these networked wrinkles.
Does anyone have any thoughts as to how I might avoid this problem, before I go insane!!
Dear Sirs: I am working on the sputtering NiFe/Mo magnetic multilayers by using HREM. The interface between the NiFe and Mo layers may be a kind of NiFeMo alloy, but I can't make decision about the idea. So I hope to know the NiFeMo phase diagram and the solution degree between NiFe and Mo.
I am looking for the help from you and please tell me the references where I can find the NiFeMo phase diagram.
The Department of Biological Sciences of the Exacts and Natural Sciences Faculty of the Buenos Aires University are looking for donation of a TEM and-or SEM equipment, not older than 15 years and still woorking. We will paid any handling and shipping cost. It would be very helpfull for us any other kind of assistance. Thank you vey much in advance.
Please respond to me at the addresses and numbers listed below.
Gabriel Adriano Rosa Area Microscopia Electronica, Depto. Cs. Biologicas Fac. de Ciencias Exactas y Naturales, Universidad de Buenos Aires Ciudad Universitaria, 4 piso, Pab. II, CP 1428, Buenos Aires, ARGENTINA
Z.Zhang's Postgraduate students wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear Sirs: } I am working on the sputtering NiFe/Mo magnetic multilayers by } using HREM. The interface between the NiFe and Mo layers may be a kind } of NiFeMo alloy, but I can't make decision about the idea. So I hope to } know the NiFeMo phase diagram and the solution degree between NiFe and } Mo. } } I am looking for the help from you and please tell me the } references where I can find the NiFeMo phase diagram. } } Yours Sincerely } } Gao Yihua
Gao Yihua,
Try with ASM book of Phase Diagrams. More info about this book you may found at: http://http://www.ASM-INTL.ORG. They have single phase diagram delivery service.
"Frustration" is a kind word for it. I fought for 2 whole years with a project requiring embedding of skin (muscle and epithelium). It was one of the worst fights I ever conducted in my association with LM - TEM. I finally was able to solve it and wrote a paper - Crowley, Hildegard, H., Elimination or Reduction of Wrinkles in Semithin Epoxy Sections.....1988.Stain Technology, Vol 64, No. 5, p 221. From your short description I surmise that you need to change your protocol, starting with dehydration (perhaps earlier). You need to change your embedding medium. Araldite-Epon-DDSA is "too soft" or not crosslinked enough for your application. The differences in viscosities between Araldite and Epon is so great that muscle tissue may act as a diffusion barrier to the Araldite, thus allowing the seperation of monomers before polymerization. I personally would abondon that formulation for that purpose, but if there were reason not to, I would take other measures. Please call me if you cannot get the paper or have further questions. I understand your frustration completely! I cannot go into a very lengthy discussion at this time, but I suggest for a trial the following: Take some of your wrinkled sections, soak the slides in water for 30 minutes or so, dry off the slides with tissue paper. Place slides in a rack. Outfit a vaccum jar with Na pentoxide ( a flat dish of some sort - use a generous amount). Put the dish in the bottom of the jar and cover the dish with a piece of large filter paper. Put the inset of the vaccum jar back in place. Set the rack of slides on the inset (rack of slides will be elevated over the pentoxide). Grease the rim of the lid well and close. Gradually pull a good vaccum on the jar. Leave the slides for 48 hours. Gradually admit air to the jar. Inspect. It may not work, but it is worth a try. If it works, you are in clover. If not, call me. 303-871-3026. Do not get discouraged. You CAN solve this problem. Bye, Hildy Crowley University of Denver
What do routine labs do with uranyl acetate waste. Ours has been picked up by in house safety department and is being stored in drums as there isn't a company that will pick up to discard. Is anyone else having this problem. It is a .025mg uranyl acetate to 100 ml of 50% ethanol washed out with copious amounts of sterile distilled water. Any imput will be appreciated. Many thanxs
} My SEM books are quite vague regarding what channeling contrast on the } SEM is. Can anyone out there tell me } 1) What is the mechanism of channeling contrast on the SEM? Basically, the BSE yield is dependant on the relative beam/crystal orientation for a sample, with ``large'' changes in BSE yield occourring over small changes in angle near the Bragg conditions for the various crystal planes in the sample.
} 2) What detectors/equipment is necessary to do channeling contrast on } the SEM? Channeling contrast generally is on the order of only a few percent, so maximization of signal is important. High BSE collection efficiencies are desirable, as well as a large probe current. Since the signal yield is a function of the electron trajectory relative to the crystal, a small convergence angle for the scanning probe is also a good idea.
} 3) What are the sample and sample prep. requirements for channeling } contrast on the SEM? -A ``smooth'' surface, since the ECC signal is easily swamped by topographic contrast. Likewise, a sample of farily uniform composition is also helpful beceause of Z contrast. -A clean, undisturbed crystal surface. This is beceause ECC requires a coherent electron beam interacting with the crystal, and coherency is lost over a fairly short distance of travel through any surface layers. For both these reasons, I prepare most of my channeling contrast samples by electropolishing.
Note: if channeling contrast is being used for grain boundries and the like, the contrast can often be picked up by a secondary or E-T style detector beceause of the linkage between BSE yield and secondary yield. A further note: ECC for imaging crystal defects follows all the points above, with the addition that the sample must be oriented relative to the beam in a fashion strongly analogous to orienting a TEM sample for diffraction contrast. This usually requires either large crystals or the ability on your microscope to do selected area channeling, so that the desired channeling condition can be established. } Have fun with ECC! Ben Simkin, simkin-at-egr.msu.edu Michigan State University dept. Materials Science
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Sirs: I am working on the sputtering NiFe/Mo magnetic multilayers by using HREM. The interface between the NiFe and Mo layers may be a kind of NiFeMo alloy, but I can't make decision about the idea. So I hope to know the NiFeMo phase diagram and the solution degree between NiFe and Mo.
I am looking for the help from you and please tell me the references where I can find the NiFeMo phase diagram.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Sirs: I am working on the sputtering NiFe/Mo magnetic multilayers by using HREM. The interface between the NiFe and Mo layers may be a kind of NiFeMo alloy, but I can't make decision about the idea. So I hope to know the NiFeMo phase diagram and the solution degree between NiFe and Mo.
I am looking for the help from you and please tell me the references where I can find the NiFeMo phase diagram.
Hello all, I need to look at the surface morphology of some yeast cells and as I'm a physics sort of person I need a little help.
I going to be using an ESEM, but what is the best way to prepare yeast cells for this sort of examination?
A step by step answer or reference would be great.
many thanks
David
Dr. David C. Bell Room 13-1018 E-Mail: dcb-at-MIT.EDU Center for Mat. Sci. and Eng. PH: (617) 253-3317 Massachusetts Institute of Technology FAX: (617) 258-6478 77 Massachusetts Ave, Cambridge, MA 02139-4307
Web Promotion Spider uses artificial intelligence to give your web page(s) optimum search engine placement. Get the jump on the compotition!! Your Site Traffic depends on your search engine placement. The sites on the first page of a search engine get ten times more hits than the sites on the third page - the Web Promotion Spider will increase your Site Hits!! Download a demo version to check out the Spider's features. http://www.marketcom.com
I am looking for the best options on the market for hardware and software to create video presentations. I want to be able to create video from series of TIFF images (or whatever format one uses) and edit these videos, add text and effects of different kinds...
I would appreciate relevant comments from you about: - Hardware for this that goes with a PC - Software for this that runs under Windows NT 4.0
I am simply interested to hear what you may use for this and comments about it.
Thanks, Martin
----------------------------------------------- Martin K=F6hler Department of Molecular Medicine Rolf Luft Center for Diabetes Research L6B:1 Karolinska Hospital S-171 76 STOCKHOLM SWEDEN phone 46-8-51775732 46-8-51775727 fax 46-8-51773658 E-mail mk-at-enk.ks.se ---------------------------------------------
Under ideal (and reasonably attainable conditions), what is the detection limit (in grams) for EDX and WDX? Thanks.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901-4402 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 Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } ----------------------------- } -----------------------------------------. } } Under ideal (and reasonably attainable conditions), what is the } detection } limit (in grams) for EDX and WDX? Thanks. } } ...
The question you ask is not specific enough ... to many factors to be considered with regard to which elements you are interested in, and (e.g.) ... how sensitive your specimen is to long count times and high beam currents ... ... ask again ...
cheerios, shAf -- ~~~~~~~~~~~~~~~~~~~~~~~ cogito, ergo zZOooOM ~~~~~~~~~~~~~~~~~~~~ Michael Shaffer - Geological Sciences - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
I need a solution to electro polishing a Cu, Ni, Al alloy (~ 80:14:4 wt%)for EBSP work. A CrO3 + HClO4 has been suggested, however does anyone know of any other suitable solutions.
Thanks in advance.
Regards,
Keith.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Dr. K. Moulding.
Materials Characterisation and Preparation Facility Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong.
Postdoctoral Studentship in Surface Science and Engineering
Applications are invited for Postdoctoral Research Studentships in the following subjects of study:
- micro and nanotribology - study of laser processed materials using transmission electron microscopy.
The Studentships are tenable for one year, renewable for a second year and include a maintenance grant of about US$20,000 per year. The Studentship will be supported by a research contract. Applicants should have a Ph.D. degree in an appropriate subject, or expect to obtain it before receiving the grant. Experience in transmission electron microscopy of metallic materials or ceramics is essential. Formal applications including a CV, names of two referees and specification of areas of interest and past experience should be sent to:
Prof. R. Vilar Departamento de Engenharia de Materiais Instituto Superior Ticnico Av. Rovisco Pais, 1096 Lisboa Codex, Portugal Tel. no. 351-1-8418121, fax no. 351-1-8418121 Email address: pcrvilar-at-alfa.ist.utl.pt.
New additions have been posted at http://members.aol.com/ImagProcTK/updates.htm for both Mac and Windows users. The Measure/Size&Shape routine previously available for Mac is now provided for Windows. Routines to draw grids of lines (horizontal or vertical, or radial with either uniform or sine weighting) are useful for stereological counting or to AND with binary images for measurement (both platforms). Coming next month: Measure/Select for Windows, Color space conversion routines, and more...
} } } Hello all, } I need to look at the surface morphology of some yeast } cells and as I'm a physics sort of person I need a little help. } } I going to be using an ESEM, but what is the best way to } prepare yeast cells for this sort of examination? } } A step by step answer or reference would be great. }
Don't prepare the samples! Mount them on a peltier stage stub, cool them, get the chamber water vapour stabilised and view them as they are. For anything more detailed please mail me direct.
Chris
Chris Gilpin Biological Sciences Electron Microscope Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 161 275 5170 fax +44 161 275 5171 http://www.biomed.man.ac.uk/biology/emunit/emhome.html
Greetings, OK, this sounds a bit off topic, but honestly, I encountered this problem in a procotol for extracting polymerized butyl-methylmethacrylate resin from sections. The protocol, from the early 1950's, simply calls for "amyl alcohol" and "amyl acetate". Nowadays, there is a very popular solvent called "ISOamyl alcohol" (or more formally, 3methyl-1butanol), and I think this might have been what was being refered to in the fifties as simply amyl alcohol. Contrarywise, there is a solvent that is still called in the catalogs "amyl acetate" (more formally, pentacetate), that I guess is what was meant (despite the fact that a compound exists that is now called "ISOamyl acetate". I would like to try this protocol, and I would rather not all of the different amyl alcohols (n-, t-, dlsec- etc). Can anyone with a long memory or a good chemistry background help here? Many thanks! Tobias Baskin
Don't know if this will work for your alloy, but I have used a simple electrolyte that you can use at room temperature for jet polishing various copper alloys for TEM. Might work for your purposes.
25% Phosphoric Acid 25% Ethylene Glycol 50% Distilled Water
Good Luck
Dan Edwards
------------------------------------------------------- Dan Edwards Structural Materials Research Section Battelle Pacific Northwest National Laboratory P.O. Box 999, MSIN P8-15 Richland, WA 99352
} -----Original Message----- } From: Keith Moulding [SMTP:mcmouldk-at-uxmail.ust.hk] } Sent: Tuesday, September 02, 1997 12:06 AM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Electro polishings of Cu,Ni,Al alloy } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } ---------------------------------------------------------------------- } -. } } Hello, } } I need a solution to electro polishing a Cu, Ni, Al alloy (~ 80:14:4 } wt%)for } EBSP work. A CrO3 + HClO4 has been suggested, however does anyone } know of } any other suitable solutions. } } Thanks in advance. } } Regards, } } Keith. } } } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Dr. K. Moulding. } } Materials Characterisation and Preparation Facility } Hong Kong University of Science and Technology, } Clear Water Bay, } Kowloon, } Hong Kong. } } FAX: (852) 2358 2451 } TEL: (852) 2358 8724 } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
There is a Fe-Mo-Ni ternary phase diagram in the Journal of Phase Equilibria 15 (6), 622-626. The article includes references to other papers. } } Dear Sirs: } I am working on the sputtering NiFe/Mo magnetic multilayers by } using HREM. The interface between the NiFe and Mo layers may be a kind } of NiFeMo alloy, but I can't make decision about the idea. So I hope to } know the NiFeMo phase diagram and the solution degree between NiFe and } Mo. } } I am looking for the help from you and please tell me the } references where I can find the NiFeMo phase diagram. } } Yours Sincerely } } Gao Yihua
Russell E. Cook Scientific Associate Electron Microscopy Center for Materials Research Argonne National Laboratory Building 212 9700 South Cass Avenue Argonne, IL 60439 (630)252-7194 FAX: (630)252-4798
AMT has a "fast track" opportunity for a national field service engineer. The position requires understanding of electron microscopes, digital technology and software. This individual will be responsible for installation and support of AMT's imaging and motion control products for electron microscopy.
Inquiries and questions should be addressed to:
Jim Mancuso Advanced Microscopy Techniques Corp. 3 Electronics Avenue Danvers MA 01923
Acording to my material safety data sheets: Amyl alcohol is 1-pentanol, CAS# 71-41-0, although the name is also used for a mixture. Isoamyl alcohol is 3methyl-1butanol, CAS #123-51-3. Amyl acetate is pentacetate or 1-pentyl acetate or N-pentyl acetate or N-amyl acetate. CAS# 628-63-7. Isoamyl acetate is 3methyl-1butyl acetate or isopentyl acetate, CAS# 123-92-2. } } Greetings, } OK, this sounds a bit off topic, but honestly, I encountered this } problem in a procotol for extracting polymerized butyl-methylmethacrylate } resin from sections. The protocol, from the early 1950's, simply calls for } "amyl alcohol" and "amyl acetate". Nowadays, there is a very popular } solvent called "ISOamyl alcohol" (or more formally, 3methyl-1butanol), and } I think this might have been what was being refered to in the fifties as } simply amyl alcohol. Contrarywise, there is a solvent that is still called } in the catalogs "amyl acetate" (more formally, pentacetate), that I guess } is what was meant (despite the fact that a compound exists that is now } called "ISOamyl acetate". } I would like to try this protocol, and I would rather not all of } the different amyl alcohols (n-, t-, dlsec- etc). Can anyone with a long } memory or a good chemistry background help here? } Many thanks! } Tobias Baskin } } } _ ____ ^ __ ____ Tobias I. Baskin } / \ / / \ / \ \ University ofMissouri } / | / / \ \ \ BiologicalSciences } /___/ /__ /___ \ \ \__ 109 Tucker Hall } / / / \ \ \ Columbia, MO 65211-7400 USA } / / / \ \ \ voice: 573-882-0173 } / /____ / \ \__/ \____ fax: 573-882-0123
Russell E. Cook Scientific Associate Electron Microscopy Center for Materials Research Argonne National Laboratory Building 212 9700 South Cass Avenue Argonne, IL 60439 (630)252-7194 FAX: (630)252-4798
Traditionally, AMYL ALCOHOL is a mixture of 3-methyl-1-butanol and 2-methyl-1-butanol, derived from the distillation of fermented STARCH (Latin: amylum). This is almost certainly what was meant in the 1950s and even much later. These days, you are likely to find this listed as iso-amyl or iso-pentyl alcohol (or acetate). When catalogues list the n-isomer they generally specify n-amyl or n-pentyl.
For any traditional protocol, the iso-amyl (or pentyl) is what would have been on the shelf at the time.
Useful source: "Chemistry of Organic Compounds" by Noller - the 2nd edition (1957) is particularly good for historical information.
(Incidentally, the higher alcohols are not formed by side-reactions on the glucose from the starch, but are derived from amino-acids derived from the yeast or present in the potatoes, or whatever).
+------------------------------------------------------------------------+ | 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 need some advice and wonder if there is any one who can help me. We have a Hitachi H-600 tem, the machine is quite old ie. 16 years. A collegue from a neighbouring centre's, Hitachi h-600 is giving problems. They would like to borrow our high voltage board and test it on their machine to make sure that the board is not broken. I am in a dilemma as I don't know if what ever caused thei.r problem will damage our board. I feel I would like to help them out but not if there is a chance that I might be making problems for myself. I would really appreciate any advice as I have not been running our EM lab for very long (2 years), and am still not 100% confident with the machine.(All our senior staff left and we have been unable to replace them, so I don't really have anyone to ask who has a lot of experience with the mechanical running of the Hitachi)
Thanks Helen Ilsley Diagnostic EM Lab UCT Medical School Groote Schuur Hospital Cape Town South Africa e-mail : hilsley-at-chempath.uct.ac.za
The question of detection limits in X-ray spectroscopic analysis is a complex one which generally does not have a simple answer. This matter is discussed in some detail by Heinrich in his book 'Electron Beam X-ray Micro Analysis', Van Nostrand-Reinhold, 1981 (ISBN 0-442-23286-1) p. 193 & p. 216, with the conclusion that the term 'm inimum detectability limit' probably should not be used.
The matter is also discussed by Goldstein, et. al. in their book 'Scanning Electron Microscopy and X-ray Microanalysis', Plenum Prewss,2nd Ed., 1992, (ISBN 0-306-44175-6). In Table 9.17, p. 501, they give calculated values comparing MDLs for several different elements for both the EDS and WDS detection systems. If you don't have this book (you should, however, by all means obtain a copy if you are doing any work in SEM of X-ray microanalysis) I can fax you a copy of this discussion.
Best regards,
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Looking for a substitute for methanol for use in electropolishing of metal samples for TEM observation. Additionally it should have a flash point near or above 100 deg. F.
Hi John and Michael et al: I recall that one of your Presidents (Kennedy?) was looking for a one handed science adviser; the ones he had always prevaricated "on the one hand and on the other hand".
Michael is quite right, detection limits vary greatly depending on endless factors. However, it is useful to have some figures as guidelines: Considering say the 10 elements following sodium. Detection of these is about best. For these in EDX the limit for quantitative analysis is about 1%. Detection limit is about 0.1%. Increasing counts and counting times beyond the customary 100 seconds at perhaps 2000cps will scarcely improve either limit.
WDX is near quantitative to its detection limit and that is at least two orders of magnitude greater than is EDX.
I'll enter correspondence only when it concerns errors in excess of five orders of magnitude. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 400+ Links, MSDS ************************ http://www.proscitech.com.au
} John J. Bozzola wrote: } } } } Under ideal (and reasonably attainable conditions), what is the } } detection } } limit (in grams) for EDX and WDX? Thanks. } } The question you ask is not specific enough ... to many factors to be } considered with regard to which elements you are interested in, and } (e.g.) ... how sensitive your specimen is to long count times and high } beam currents ... ... ask again ... } cheerios, shAf } -- } ~~~~~~~~~~~~~~~~~~~~~~~ cogito, ergo zZOooOM ~~~~~~~~~~~~~~~~~~~~ } Michael Shaffer - Geological Sciences - University of Oregon } mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
Dear Teresa - I waited a couple of days but there was no reply on the listserver. Here is mine: I do not know about your regulations, maybe you must collect the stuff and then dispose of it "properly". But too many regulations are not wise - eg. the dangerous goods shipping laws which appear designed to make things more expensive but not safer.
Uranium occurs in trace amounts throughout the environment, including seawater and especially in granite. U does not concentrate in the food chain like P, K, Pb, Hg or some organic compounds. For very small quantities, as are used in EM labs, prompt disposal with some water into the sewage system appears perfectly reasonable to me. The mistake is to store the stuff and to accumulate larger quantities.
---------- } From: Flores, Teresa {tflore-at-lsumc.edu} } To: Microscopy Newsgroup {Microscopy-at-sparc5.microscopy.com} } Subject: } Date: Saturday, 30 August 1997 4:33
} } What do routine labs do with uranyl acetate waste. Ours has been picked up } by in house safety department and is being stored in drums as there isn't a } company that will pick up to discard. Is anyone else having this problem. } It is a .025mg uranyl acetate to 100 ml of 50% ethanol washed out with } copious amounts of sterile distilled water. Any imput will be appreciated. } Many thanxs } }
Anyone out there with experience exporting a Tencor P-11 Profilometer 3D data set to an external analysis program (e.g. IGOR or Nanoscope AFM)? We are looking for suggestions on successful transfers.
Michael T. Dineen The Dow Chemical Company 1897 Building Midland MI 48667 (517/636-4008 4 517/638-6443 + mtdineen-at-dow.com
Hello, I need to replace the 12 inch bell jar on a Denton 502. Denton wants $627 for a new one. Does any one know a source of used bell jars, etc. for this equipment? Thanks Wallace Ambrose
Jim, U in the USA must be disposed of in a way other than "down the sewer". It may occur in trace amounts in "nature" but the concentrations we work with are much higher than that. Besides the legal problems there are the moral responsibilities to not unnecessarily endanger others with the chemicals we use. True, uranium is one of the milder toxins we handle....if handled properly, but it still is a beta emitter and can do extensive tissue damage if allowed to come into contact with any soft tissues. Flushing this waste down the sewer does not guarantee that this will not happen to some unsuspecting individual. That is why there are regulations in place for the proper handling and disposal of this chemical. I also empathize with Teresa's dilema because we are in the same position....radiation waste handlers won't take it because it is a naturally occurring isotope and hazardous waste handlers won't take it because it is radioactive....a perfect catch 22. If anyone can come up with a practical solution to this dilemma that satisfies the current clean water act regulations we would be more than happy to listen.
Jim Darley wrote: } Dear Teresa - } I waited a couple of days but there was no reply on the listserver. Here is } mine: } I do not know about your regulations, maybe you must collect the stuff and } then dispose of it "properly". But too many regulations are not wise - eg. } the dangerous goods shipping laws which appear designed to make things more } expensive but not safer. } } Uranium occurs in trace amounts throughout the environment, including } seawater and especially in granite. U does not concentrate in the food } chain like P, K, Pb, Hg or some organic compounds. For very small } quantities, as are used in EM labs, prompt disposal with some water into } the sewage system appears perfectly reasonable to me. } The mistake is to store the stuff and to accumulate larger quantities. } } ---------- } } From: Flores, Teresa {tflore-at-lsumc.edu} } } To: Microscopy Newsgroup {Microscopy-at-sparc5.microscopy.com} } } Subject: } } Date: Saturday, 30 August 1997 4:33 } } } } } What do routine labs do with uranyl acetate waste. Ours has been picked } up } } by in house safety department and is being stored in drums as there isn't } a } } company that will pick up to discard. Is anyone else having this problem. } } It is a .025mg uranyl acetate to 100 ml of 50% ethanol washed out with } } copious amounts of sterile distilled water. Any imput will be } appreciated. } } Many thanxs } } } }
I agree, EDX detection limit somewhere just less than 1%...but
I work with polymer fibers and by ashing I can get great analysis of additives at 100 ppm in the polymer. This doesn't change the EDX detection limit per se, but there is more than one way to skin a cat.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi John and Michael et al: I recall that one of your Presidents (Kennedy?) was looking for a one handed science adviser; the ones he had always prevaricated "on the one hand and on the other hand".
Michael is quite right, detection limits vary greatly depending on endless factors. However, it is useful to have some figures as guidelines: Considering say the 10 elements following sodium. Detection of these is about best. For these in EDX the limit for quantitative analysis is about 1%. Detection limit is about 0.1%. Increasing counts and counting times beyond the customary 100 seconds at perhaps 2000cps will scarcely improve either limit.
WDX is near quantitative to its detection limit and that is at least two orders of magnitude greater than is EDX.
I'll enter correspondence only when it concerns errors in excess of five orders of magnitude. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 400+ Links, MSDS ************************ http://www.proscitech.com.au
} John J. Bozzola wrote: } } } } Under ideal (and reasonably attainable conditions), what is the } } detection } } limit (in grams) for EDX and WDX? Thanks. } } The question you ask is not specific enough ... to many factors to be } considered with regard to which elements you are interested in, and } (e.g.) ... how sensitive your specimen is to long count times and high } beam currents ... ... ask again ... } cheerios, shAf } -- } ~~~~~~~~~~~~~~~~~~~~~~~ cogito, ergo zZOooOM ~~~~~~~~~~~~~~~~~~~~ } Michael Shaffer - Geological Sciences - University of Oregon } mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
I am looking for a set of circuit diagrams for a Zeiss EM-10A transmission electron microscope. If anyone is willing to part with an old set, or will allow me to copy a set, please email me at:
One of our customers has asked for some advice. He's picking up 1 inch diameter, .040" thick aluminum discs and exposing them to a chemical process which requires the least amount of the disc to be obstructed. He now uses triceps to hold the discs but they lose their memory after a mumber of uses. Can anyone suggest a better way to handle the discs?
Thanks, John Arnott Ladd Research ladres-at-worldnet.att.net
Teresa, I was hoping that someone experianced in haz. waste disposal methods would comment on your question. In my experiance, the problem with UAC waste disposal is if the radioactive material is in methanol. I have no trouble having the waste picked up if the UAC is in water. I have been told by Environmental health and Safety personnel that no disposal option exists for the UAC in methanol. You have brought up a very important topic and hopefully comments from other labs and especially from individuals experianced in waste disposal may follow.
Robert Cox Shriner Hospital Galveston Tx. -------- -----------------------------------------------------------------------.
Dear Teresa - I waited a couple of days but there was no reply on the listserver. Here is mine: I do not know about your regulations, maybe you must collect the stuff and then dispose of it "properly". But too many regulations are not wise - eg. the dangerous goods shipping laws which appear designed to make things more expensive but not safer.
Uranium occurs in trace amounts throughout the environment, including seawater and especially in granite. U does not concentrate in the food chain like P, K, Pb, Hg or some organic compounds. For very small quantities, as are used in EM labs, prompt disposal with some water into the sewage system appears perfectly reasonable to me. The mistake is to store the stuff and to accumulate larger quantities.
---------- } From: Flores, Teresa {tflore-at-lsumc.edu} } To: Microscopy Newsgroup {Microscopy-at-sparc5.microscopy.com} } Subject: } Date: Saturday, 30 August 1997 4:33
} } What do routine labs do with uranyl acetate waste. Ours has been picked up } by in house safety department and is being stored in drums as there isn't a } company that will pick up to discard. Is anyone else having this problem. } It is a .025mg uranyl acetate to 100 ml of 50% ethanol washed out with } copious amounts of sterile distilled water. Any imput will be appreciated. } Many thanxs } }
The problem of Ur disposal will continue to exist until someone develops a plan on how to dispose of the material AND makes a decision that is legally binding. One way to start the ball rolling is to formulate some possible ways of dealing with the material and then to forward the package to appropriate legislators.
Here is what I propose: take some epoxy resin and coat the inside of a polypropylene beaker with the epoxy by building up layers and polymerizing the layers. If one uses outdated, quite viscous epoxy monomers, one can achieve several mm of thickness in a couple days. Now, use the epoxy coated vessel as a waste container into which you pour your uranium containing liquids. Keep the vessel in a warm oven or fume hood (if volatiles are involved) to speed up evaporation. Allow the uranium liquids to evaporate to dryness (taking care to avoid generating dust) and apply another layer of epoxy over the uranium salts. What one gets is a layered system of epoxy/uranium/epoxy/uranium etc. Keep this up until the polypropylene vessel is completely filled with epoxy. The enrobed uranium can then be disposed in a toxic waste burial site where it might be further enrobed.
If users scale back on the use of uranium using tiny amounts and minimize rinse volumes, some of the problem will already be solved.
Key ingredients: scale down use, encapsulate dried salts in epoxy.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing 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 ####################################################################
Are you replacing the bell jar because the rim is chipped ?. If so have you thought of having the rim cut down to remove the damaged area ? We had that done several times by a glass blower at a fraction of the cost of a new bell jar.
Jordi Marti ----------------------- Hello, I need to replace the 12 inch bell jar on a Denton 502. Denton wants $627 for a new one. Does any one know a source of used bell jars, etc. for this equipment? Thanks Wallace Ambrose
How about a trivet, like metalsmiths use for enamelling? (Check the metals program at your nearest college's art dept.) The tips can be modified by a little judicious filing to minimize the areas of contact between the disc and the legs of the trivet.
Phil
} To All, } } One of our customers has asked for some advice. He's picking up 1 inch } diameter, .040" thick aluminum discs and exposing them to a chemical } process which requires the least amount of the disc to be obstructed. He } now uses triceps to hold the discs but they lose their memory after a } mumber of uses. } Can anyone suggest a better way to handle the discs? } } Thanks, } John Arnott } Ladd Research } ladres-at-worldnet.att.net
I have no financial interest in VWR, etc.... } } } Harold J. Crossman } OSRAM SYLVANIA INC. } Lighting Research Center } 71 Cherry Hill Dr. } Beverly, MA 01915 } Phone: (508) 750-1717 } E-mail: crossman-at-osi.sylvania.com } } Our web sites: www.sylvania.com } www.siemens.com } -- } } "Crossman, Harold" {crossman-at-osi.SYLVANIA.com}
I have no financial interest in VWR, etc.... } } } Harold J. Crossman } OSRAM SYLVANIA INC. } Lighting Research Center } 71 Cherry Hill Dr. } Beverly, MA 01915 } Phone: (508) 750-1717 } E-mail: crossman-at-osi.sylvania.com } } Our web sites: www.sylvania.com } www.siemens.com } -- } } "Crossman, Harold" {crossman-at-osi.SYLVANIA.com}
Uranium in the USA must be disposed of in a way other than "down the sewer". It may occur in trace amounts in "nature" but the concentrations we work with are much higher than that. Besides the legal problems there are the moral responsibilities to not unnecessarily endanger others with the chemicals we use. True, uranium is one of the milder toxins we handle....if handled properly, but it still is a beta emitter and can do extensive tissue damage if allowed to come into contact with any soft tissues. Flushing this waste down the sewer does not guarantee that this will not happen to some unsuspecting individual. That is why there are regulations in place for the proper handling and disposal of this chemical. I also empathize with Teresa's dilema because we are in the same position....radiation waste handlers won't take it because it is a naturally occurring isotope and hazardous waste handlers won't take it because it is radioactive....a perfect catch 22. If anyone can come up with a practical solution to this dilemma that satisfies the current clean water act regulations we would be more than happy to listen.
Jim Darley wrote: } Dear Teresa - } I waited a couple of days but there was no reply on the listserver. Here is } mine: } I do not know about your regulations, maybe you must collect the stuff and } then dispose of it "properly". But too many regulations are not wise - eg. } the dangerous goods shipping laws which appear designed to make things more } expensive but not safer. } } Uranium occurs in trace amounts throughout the environment, including } seawater and especially in granite. U does not concentrate in the food } chain like P, K, Pb, Hg or some organic compounds. For very small } quantities, as are used in EM labs, prompt disposal with some water into } the sewage system appears perfectly reasonable to me. } The mistake is to store the stuff and to accumulate larger quantities. } } ---------- } } From: Flores, Teresa {tflore-at-lsumc.edu} } } To: Microscopy Newsgroup {Microscopy-at-sparc5.microscopy.com} } } Subject: } } Date: Saturday, 30 August 1997 4:33 } } } } } What do routine labs do with uranyl acetate waste. Ours has been picked } up } } by in house safety department and is being stored in drums as there isn't } a } } company that will pick up to discard. Is anyone else having this problem. } } It is a .025mg uranyl acetate to 100 ml of 50% ethanol washed out with } } copious amounts of sterile distilled water. Any imput will be } appreciated. } } Many thanxs } } } }
The Duniway Stockroom Corp., 1305 Space Park Way, Mountain View, CA 94043, Tel. 800-446-8811, e.mail: info-at-duniway.com, deals in new, used, and rebuilt vacuum equipment. You might copntact them.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
} Date: Thu, 04 Sep 1997 14:11:03 +0200 } From: Catherine Goffaux {catherine.goffaux-at-wkb.be} } To: njoschko-at-datagram.be, claudejp-at-esvax.dnet.dupont.com, orion-at-infoboard.be, } deschuyt-at-sbbio.be, bdd.translations-at-skynet.be, labio-at-telecom-plus.sn, } sandra.rens-at-vulcan.be } Subject: (Fwd) (Fwd) Virus Warning (fwd) -RETRANSMIS } } Received: from mserv.rug.ac.be by wkb.be (SMI-8.6/SMI-SVR4) } id NAA01015; Sun, 31 Aug 1997 13:12:47 +0200 } Received: from eduserv2.rug.ac.be by mserv.rug.ac.be with SMTP id AA14051 } (5.67b/IDA-1.5 for {inge.vanderhaegen-at-wkb.be} ); Sun, 31 Aug 1997 13:21:01 +0200 } Received: from localhost by eduserv2.rug.ac.be (SMI-8.6/SMI-SVR4) } id NAA03840; Sun, 31 Aug 1997 13:20:59 +0200 } Message-Id: {Pine.SOL.3.94.970831131452.3715A-100000-at-eduserv2.rug.ac.be} } Date: Sun, 31 Aug 1997 13:20:59 +0200 } From: Kristof Van der Haegen {Kristof.VanderHaegen-at-rug.ac.be} } To: Karl.Theeten-at-rug.ac.be } Cc: Peter.Forret-at-keyware.be,Bartel.Van.Der.Haegen-at-mobile.belgacom.be, } Benedikt.Ameloot-at-rug.ac.be, Gunther.Heene-at-rug.ac.be, } Luc.VanSintJan-at-rug.ac.be, Samuel.VanBelle-at-rug.ac.be, } inge.vanderhaegen-at-wkb.be } Subject: (Fwd) (Fwd) Virus Warning (fwd) } Mime-Version: 1.0 } Content-Type: text/plain } Content-Disposition: inline } } PLEASE read the following warning. } WARNING!!!!!! If you receive an e-mail titled "JOIN THE CREW" } DO NOT open it! } It will erase EVERYTHING on your hard drive! Send this letter out to } as many people you can.......this is a new virus and not many people } know about it! } } This information was received this morning from IBM, please share it } with anyone that might access the Internet. } } Also, } } If anyone receives mail entitled; PENPAL GREETINGS! please delete it } WITHOUT reading it!! This is a warning for all Internet users - } there is a dangerous virus propagating across the Internet through an } e-mail message entitled "PENPAL GREETINGS!". } } DO NOT DOWNLOAD ANY MESSAGE ENTITLED "PENPAL GREETINGS"!! } } This message appears to be a friendly letter asking you if you are } interested in a penpal, but by the time you read this letter, it is } too late. The trojan horse" virus will have already infected the boot } sector } of your hard drive, destroying all of the data present. It is a } self-replicating virus, and once the message is read, it will } AUTOMATICALLY forward itself to anyone who's e-mail address is present } in YOUR mailbox! } } This virus will DESTROY your hard drive, and holds the potential to } DESTROY the hard drive of anyone whose mail is in your in box, and } who's } mail is in their in box and so on. If this virus keeps getting } passed, it has the potential to do a great deal of DAMAGE to computer } networks worldwide!!!! } } Please, delete the message entitled "PENPAL GREETINGS!" as soon as you } see it! And pass this message along to all of your friends, relatives } and the other readers of the newsgroups and mailing lists which you } are on so that they are not hurt by this dangerous virus!!!! } } Please pass this along to everyone you know so this can be stopped. } PASS THIS ON TO YOUR FRIENDS!!! WARNING !!! } There is a new virus going around in the last couple of days!!! } DO NOT open or even look at any mail that you get that says: "Returned } or Unable to Deliver" This virus will attach itself to your computer } components and render them useless. Immediately delete any mail items } that says this. AOL has said this is a very dangerous virus, and there } is NO remedy for it at this time, Please Be Careful, And forward to } all your on-line friends A.S.A.P. } } Forward this A.S.A.P. to every single person you know!!!!!!!!! } *************************************************** } } } }
Best regards,
Paul Vanderlinden. Sales Manager.
======================================================================= See our web site: http://www.microscopy-uk.org.uk
Jim Darley's posting is hardly furthering good science.
1: Detection limits (John's original question). Modern EDX systems are easily capable of quantitative analysis in the sub-1wt% range. See, for example http://prism.mit.edu/facltis/stem/stmexam.htm (the second illustration on that page) where I was getting analyses for Cr in steel of the order of 0.6+/-0.1 wt%. I don't know about you, but I consider this quantitative. This, mind you, was in a measurement where I was attempting to optimize spatial resolution rather than sensitivity, and the acquisition time was 60sec. per data point. This example, of course, was from a STEM. There are many more examples in the literature.
Using beam gating techniques such as those developed by Charlie Lyman and colleagues, EDX data can readily be acquired at 20,000-40,000 counts per real second, if spatial resolution is sacrificed. Combine this with an acquisition time of 1,000 seconds (by no means unrealistic for an important measurement) and the detection limit is in the rage of, or better than, 0.01wt%.
Of course, in the SEM, which may have been the point of John's original question, the situation is not the same, the beam voltage is lower (resulting in poorer peak/bremmstrahlung ratios) and the emission of x-rays from a solid sample is different from that in a thin foil, but these are some of the variables that Michael quite rightly pointed out must be considered.
2: Comparison between EDX and WDX.
This is like comparing apples and oranges, because the instruments designed with them are generally intended for different purposes.
WDX has a much better peak resolution than WDX, which results in better measured P/B ratios (and hence improved statistics), as well as much better capability in resolving nearby x-ray lines. Also, because the x-ray counting and wavelength analysis are different functions in the crystal spectrometer, available countrates have traditionally been higher in WDX than EDX (although modern EDX detectors are an order of magnitude faster than they were fifteen years ago). Against this must be set the fact that WDX is inherently a serial technique (although multiple spectrometers help here), while EDX is a parallel technique (compare the advantages of PEELS over SEELS - although this is not a totally fair comparison). Anyway, the advantage of WDX (ignoring the capability of resolving peak overlaps) is improved statistical precision. Is this useful?
Well, maybe.
By far the most significant parameter which affects electron-induced x-ray emission spectra is sample geometry. Two extreme cases are where the sample is a thin foil (as in the STEM) where, to a first approximation, the x-ray spectrum recorded by the detector is the same as that emitted, and to a second order, it is possible to derive a reasonable thickness correction for cases where the error is small. Alternatively, when the sample has dimensions large compared with the volume irradiated by the electron beam, and has an accurately known geometry compared to the incident beam and the detector (such as a polished flat sample in the microprobe), correction programs such as ZAF can do a reasonable job of extracting a quantitative analysis.
What about where the sample geometry is unknown (for example, a rough surface such as might be examined in the SEM)? In that case the uncertainty in the analysis is far, far worse than any uncertainty caused by the poor statistics of the EDX spectrum, so there is no point or advantage whatever in using WDX to try to improve things, because it won't. This is why typically an SEM has an EDX detector - it is cheaper and gives just as good an analysis (except for the somewhat poorer detection limit). In the microprobe, great care is taken in polishing and mounting the sample, so the advantage of the WDX detector can be realised.
Where does this leave us? The advantage of a WDX detector over an EDX detector *ON THE SAME SAMPLE* is limited - perhaps an order of magnitude in detection limit, and, on a flat, polished sample, also perhaps approaching an order of magnitude in precision. The WDX detector can also resolve many cases where peaks would overlap in EDX. On general rough SEM samples, the only advantages of WDX are a small improvement in detection limits and the ability to resolve overlaps (which could be important if a trace element peak overlaps a major peak in the EXD spectrum.
The President's science advisors were right - it all depends. I seem to remember that one of Jim Darley's Prime Ministers (Gough Whitlam?) had to be dismissed because Parliament would not pass his budget - but does that have anything to do with Science?
Tony Garratt-Reed
} Hi John and Michael et al: } I recall that one of your Presidents (Kennedy?) was looking for a one } handed science adviser; the ones he had always prevaricated "on the one } hand and on the other hand". } } Michael is quite right, detection limits vary greatly depending on endless } factors. However, it is useful to have some figures as guidelines: } Considering say the 10 elements following sodium. Detection of these is } about best. } For these in EDX the limit for quantitative analysis is about 1%. Detection } limit is about 0.1%. Increasing counts and counting times beyond the } customary 100 seconds at perhaps 2000cps will scarcely improve either } limit. } } WDX is near quantitative to its detection limit and that is at least two } orders of magnitude greater than is EDX. } } I'll enter correspondence only when it concerns errors in excess of five } orders of magnitude. } Cheers } Jim Darley } } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Phone +61 77 740 370 Fax: +61 77 892 313 } Great microscopy catalogue, 400+ Links, MSDS } ************************ http://www.proscitech.com.au } } } John J. Bozzola wrote: } } } } } } Under ideal (and reasonably attainable conditions), what is the } } } detection } } } limit (in grams) for EDX and WDX? Thanks. } } } } The question you ask is not specific enough ... to many factors to be } } considered with regard to which elements you are interested in, and } } (e.g.) ... how sensitive your specimen is to long count times and high } } beam currents ... ... ask again ... } } cheerios, shAf } } -- }
Anthony J. Garratt-Reed MIT Room 13-1027 77 Massachusetts Avenue Cambridge, MA 02139-4307 United States of America
We received an JEOL 6400 SEM without an instruction manual. If anyone can help us out with a copy of a manual, we'd greatly appreciate it!
************************************************************************* Lucille A. Giannuzzi, Ph.D.
Assistant Professor Dept. of Mechanical, Materials, and Aerospace Eng.
Director, Cirent/UCF Materials Characterization Facility President, Florida Microscopy Society (a local affiliate of MSA)
University of Central Florida phone (407) 823-5770 PO Box 162450 fax (407) 823-0208 4000 Central Florida Blvd. email lag-at-pegasus.cc.ucf.edu Orlando, FL 32816-2450 USA *************************************************************************
Before sending chills of panic up and down the spines of all of us computer freaks by posting information regarding an alleged virus one might check the latest information compiled by the Department of Energy at their web site dedicated to computer security.
The URL is http://ciac.llnl.gov/
The "viruses" listed are hoaxes according to CIAC.
Bob Craig OSRAM SYLVANIA Products Inc. Beverly, MA 01915
Paul Vanderlinden's posting included warnings about some computer viruses. While computer viruses can create many problems for the person or organization where an infection occurs, computer virus HOAXES can create their own problems as well.
According to the DOE, the PENPAL GREETINGS virus is a hoax, and possibly the other ones mentioned in Paul Vanderlinden's posting are also. Information about viruses can be found at the U.S. Department of Energy Computer Incident Advisory Capability (DOE-CIAC) web site which is at http://ciac.llnl.gov/ and the hoaxes page at http://ciac.llnl.gov/ciac/CIACHoaxes.html
Anyone who receives warnings about computer viruses should follow the DOE's recommended action...
"Users are requested to please not spread unconfirmed warnings about viruses and Trojans. If you receive an unvalidated warning, don't pass it to all your friends, pass it to your computer security manager to validate first. Validated warnings from the incident response teams and antivirus vendors have valid return addresses and are usually PGP signed with the organization's key." (from the DOE-CIAC web page on Internet Hoaxes)
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Try a vacuum forceps. They are available from all microscopy supply = houses and places like Edmund scientific etc. It is a pen like implement = which one puts various size needles on depending on the size one wants to = pick up. Suction is created by a very small motor. No microscopy lab = should be without one.
Judy M.
Judy Murphy, PhD Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207
Phone: 209/954-5284 FAX: 209/954-5600 e-mail: murphy-at-sjdccd.cc.ca.us program web page: http://www.sjdccd.cc.ca.us/ElectMicro/sjdc.html
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
To All,
One of our customers has asked for some advice. He's picking up 1 inch diameter, .040" thick aluminum discs and exposing them to a chemical process which requires the least amount of the disc to be obstructed. He now uses triceps to hold the discs but they lose their memory after a mumber of uses. Can anyone suggest a better way to handle the discs?
Thanks, John Arnott Ladd Research ladres-at-worldnet.att.net
------------------ RFC822 Header Follows ------------------ Received: by sjdccd.cc.ca.us with ADMIN;4 Sep 1997 14:14:35 -0800 Received: from Sparc5.Microscopy.Com (206.69.208.10) by = ms.sjdccd.cc.ca.us with SMTP (Eudora Internet Mail Server 1.2); Thu, 4 Sep 1997 14:13:51 = -0800 Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com = (8.6.11/8.6.11) id LAA02799 for dist-Microscopy; Thu, 4 Sep 1997 11:09:10 = -0500 Received: from mtigwc03.worldnet.att.net (mtigwc03.worldnet.att.net = [204.127.131.34]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id = LAA02796 for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 4 Sep 1997 11:09:09 = -0500 Received: from server ([207.116.37.66]) by mtigwc03.worldnet.att.net (post.office MTA v2.0 0613 ) with SMTP id AAA640 for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 4 Sep 1997 16:16:22 +0000 Message-ID: {340EDE5A.6A22-at-worldnet.att.net} "Paul VANDERLINDEN" {orion-at-euronet.be} X-Mailer: Mail*Link SMTP/QM 3.0.0
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Reply to: RE} (Fwd) (Fwd) Virus Warning (fwd) -RETRANSMIS
See http://ciac.llnl.gov/ciac/CIACHoaxes.html for information on the PenPal hoax.
-Mike --------------------------------------
} Date: Thu, 04 Sep 1997 14:11:03 +0200 } From: Catherine Goffaux {catherine.goffaux-at-wkb.be} } To: njoschko-at-datagram.be, claudejp-at-esvax.dnet.dupont.com, orion-at-infoboard.be, } deschuyt-at-sbbio.be, bdd.translations-at-skynet.be, labio-at-telecom-plus.sn, } sandra.rens-at-vulcan.be } Subject: (Fwd) (Fwd) Virus Warning (fwd) -RETRANSMIS } } Received: from mserv.rug.ac.be by wkb.be (SMI-8.6/SMI-SVR4) } id NAA01015; Sun, 31 Aug 1997 13:12:47 +0200 } Received: from eduserv2.rug.ac.be by mserv.rug.ac.be with SMTP id AA14051 } (5.67b/IDA-1.5 for {inge.vanderhaegen-at-wkb.be} ); Sun, 31 Aug 1997 13:21:01 +0200 } Received: from localhost by eduserv2.rug.ac.be (SMI-8.6/SMI-SVR4) } id NAA03840; Sun, 31 Aug 1997 13:20:59 +0200 } Message-Id: {Pine.SOL.3.94.970831131452.3715A-100000-at-eduserv2.rug.ac.be} } Date: Sun, 31 Aug 1997 13:20:59 +0200 } From: Kristof Van der Haegen {Kristof.VanderHaegen-at-rug.ac.be} } To: Karl.Theeten-at-rug.ac.be } Cc: Peter.Forret-at-keyware.be,Bartel.Van.Der.Haegen-at-mobile.belgacom.be, } Benedikt.Ameloot-at-rug.ac.be, Gunther.Heene-at-rug.ac.be, } Luc.VanSintJan-at-rug.ac.be, Samuel.VanBelle-at-rug.ac.be, } inge.vanderhaegen-at-wkb.be } Subject: (Fwd) (Fwd) Virus Warning (fwd) } Mime-Version: 1.0 } Content-Type: text/plain } Content-Disposition: inline } } PLEASE read the following warning. } WARNING!!!!!! If you receive an e-mail titled "JOIN THE CREW" } DO NOT open it! } It will erase EVERYTHING on your hard drive! Send this letter out to } as many people you can.......this is a new virus and not many people } know about it! } } This information was received this morning from IBM, please share it } with anyone that might access the Internet. } } Also, } } If anyone receives mail entitled; PENPAL GREETINGS! please delete it } WITHOUT reading it!! This is a warning for all Internet users - } there is a dangerous virus propagating across the Internet through an } e-mail message entitled "PENPAL GREETINGS!". } } DO NOT DOWNLOAD ANY MESSAGE ENTITLED "PENPAL GREETINGS"!! } } This message appears to be a friendly letter asking you if you are } interested in a penpal, but by the time you read this letter, it is } too late. The trojan horse" virus will have already infected the boot } sector } of your hard drive, destroying all of the data present. It is a } self-replicating virus, and once the message is read, it will } AUTOMATICALLY forward itself to anyone who's e-mail address is present } in YOUR mailbox! } } This virus will DESTROY your hard drive, and holds the potential to } DESTROY the hard drive of anyone whose mail is in your in box, and } who's } mail is in their in box and so on. If this virus keeps getting } passed, it has the potential to do a great deal of DAMAGE to computer } networks worldwide!!!! } } Please, delete the message entitled "PENPAL GREETINGS!" as soon as you } see it! And pass this message along to all of your friends, relatives } and the other readers of the newsgroups and mailing lists which you } are on so that they are not hurt by this dangerous virus!!!! } } Please pass this along to everyone you know so this can be stopped. } PASS THIS ON TO YOUR FRIENDS!!! WARNING !!! } There is a new virus going around in the last couple of days!!! } DO NOT open or even look at any mail that you get that says: "Returned } or Unable to Deliver" This virus will attach itself to your computer } components and render them useless. Immediately delete any mail items } that says this. AOL has said this is a very dangerous virus, and there } is NO remedy for it at this time, Please Be Careful, And forward to } all your on-line friends A.S.A.P. } } Forward this A.S.A.P. to every single person you know!!!!!!!!! } *************************************************** } } } }
Best regards,
Paul Vanderlinden. Sales Manager.
======================================================================= See our web site: http://www.microscopy-uk.org.uk
Course Announcement: "Optimizing Light Microscopy" When/Where: (a) New York City, November 3,1997 (b) Springfield, MA November 5, 1997 (c) Boston, MA November 7,1997 What: a lively, fast-paced slide lecture and demonstraton for anyone how uses or plans to use a light microscope: students, teachers, medical technologists, clinicians, pathologists, and lab managers. Beginning to more experienced practictioners welcome.
For details... (a) read below (b) send for brochure (c) visit the Microscopy/Microscopy Education booth at MSA - #502
Program: 1. A quick tour around the microscope - getting to know the bits & pieces 2. Koehler illumination & you: 4 critical steps for aligning you and your microscope to reduce headaches, fatigue, and errors 3. Care and cleaning 4. Useful principles for understanding and optimizing imaging 5. Putting the basics to work: a. Troubleshooting b. Understanding Phase and Hoffman Modulation Contrast 6. The Video connection: cameras, computers, and your microscope 7. Bringing out the best: quick, easy, and often free techniques for improving contrast 8. Advanced contrast techniques: Fluorescence and DIC 9. Becoming a better consumer: matching your microscope to your application 10. Questions, Answers, and Information Exchange (Note: Instructor may vary class content slightly to meet the needs of participants)
Free with your tuition: "Optimizing Light Microscopy for Biological and Clinical Laboratories" (Kendall-Hunt, 1997). Nearly 200 pages of helpful hints, quick experiments, and new iedas for getting the best from your microscope. Hot off the presses!
CEU's: 0.6 CEUs, 6 P.A.C.E CEU's Microscopy/Microscopy Education adheres to the guidelines established by the IACET.
Pricing: $150 (includes tuition, breaks, course materials, and copy of book) *****Save $25 if paid by 10/17/97.***** Send three from the same facility and save $50 on tuition for the third person.
Refund policy: Full refund for cancellations made by 10/17/97. After that date, 50% refund or full credit for future class. Substitutions accepted.
Questions: call Barbara Foster or Dr. Ken Piel at MME: (413)746-6931
Registration: Download the form below and fax to (413)746-9311 or call (413)746-6931 and ask for Ken.
Check course you will be attending: ___ New York City, November 3 (#971103) ___ Springfield, November 5 (#971105)* ___ Boston, November 7 (#971107)
Method of Payment: ____ Check enclosed for $ _______________ ____ Visa ____ Mastercard Name on credit card: __________________________________ Credit card number: ___________________________________ Expiration date: ______________________________________ ***If billing address is different from one shown above, please show billing address below: _______________________________________________________ _______________________________________________________ _______________________________________________________
The PENPAL GREETINGS! Hoax shown below appears to be an attempt to kill an e-mail chain letter by claiming that it is a self starting Trojan that destroys your hard drive and then sends copies of itself to everyone whose address in in your mailbox. Reading an e-mail message does not run it nor does it run any attachments, so this Trojan must be self starting. Aside from the fact that a program cannot start itself, the Trojan would also have to know about every different kind of e-mail program to be able to forward copies of itself to other people. This warning is totally a hoax.
FYI!
Subject: Virus Alert Importance: High If anyone receives mail entitled: PENPAL GREETINGS! please delete it WITHOUT reading it. Below is a little explanation of the message, and what it would do to your PC if you were to read the message. If you have any questions or concerns please contact SAF-IA Info Office on 697-5059.
This is a warning for all internet users - there is a dangerous virus propogating across the internet through an e-mail message entitled "PENPAL GREETINGS!". DO NOT DOWNLOAD ANY MESSAGE ENTITLED "PENPAL GREETINGS!" This message appears to be a friendly letter asking you if you are interestedin a penpal, but by the time you read this letter, it is too late. The "trojan horse" virus will have already infected the boot sector of your hard drive, destroying all of the data present. It is a self-replicating virus, and once the message is read, it will AUTOMATICALLY forward itself to anyone who's e-mail address is present in YOUR mailbox! This virus will DESTROY your hard drive, and holds the potential to DESTROY the hard drive of anyone whose mail is in your inbox, and who's mail is in their inbox, and so on. If this virus remains unchecked, it has the potential to do a great deal of DAMAGE to computer networks worldwide!!!! Please, delete the message entitled "PENPAL GREETINGS!" as soon as you see it! And pass this message along to all of your friends and relatives, and the other readers of the newsgroups and mailing lists which you are on, so that they are not hurt by this dangerous virus!!!!
Join the Crew
Circulating the Internet is an email message entitled "Join the Crew". For a virus to spread, it must be executed. Reading a mail message does not execute the mail message. Trojans and viruses have been found as executable attachments to mail messages, but they must be extracted and executed to do any harm.
CIAC still affirms that reading E-mail, using typical mail agents, can not activate malicious code delivered in or with the message.
IMPORTANT - VIRUS Alert!!!
Take note !
Someone got an email, titled as JOIN THE CREW. It has erased his hard drive. Do not open up any mail that has this title. It will erase your whole hard drive. This is a new email virus and not a lot of people know about it, just let everyone know, so they won't be a victim.
Please e-mail this to everyone you know!!! Remember the title : JOIN THE CREW
Variants of this email message are circulating the Internet. If you receive an email message entitled "Join the Crew" and it has an attachment, CIAC recommends that you delete the message and the attachment. If you receive just the message, delete the message. Please DO NOT circulate unvalidated virus alerts.
************************************************************************** Mike O'Keefe
Course Announcement: "Optimizing Light Microscopy" When/Where: "Optimizing Light Microscopy" Hosted by Providence College Dept. of Biology (Providence, RI) October 3, 1997 What: a lively, fast-paced slide lecture and demonstraton for anyone who uses or plans to use a light microscope: students, teachers, medical technologists, clinicians, pathologists, and lab managers. Beginning to more experienced practitioners welcome.
For details... (a) read below (b) send for brochure Program: 1. A quick tour around the microscope - getting to know the bits & pieces 2. Koehler illumination & you: 4 critical steps for aligning you and your microscope to reduce headaches, fatigue, and errors 3. Care and cleaning 4. Useful principles for understanding and optimizing imaging 5. Putting the basics to work: a. Troubleshooting b. Understanding Phase and Hoffman Modulation Contrast 6. The Video connection: cameras, computers, and your microscope 7. Bringing out the best: quick, easy, and often free techniques for improving contrast 8. Advanced contrast techniques: Fluorescence and DIC 9. Becoming a better consumer: matching your microscope to your application 10. Questions, Answers, and Information Exchange (Note: Instructor may vary class content slightly to meet the needs of participants. Instructor for this course: Barbara Foster of Microscopy/Microscopy Education)
Free with your tuition: "Optimizing Light Microscopy for Biological and Clinical Laboratories" (Kendall-Hunt, 1997). Nearly 200 pages of helpful hints, quick experiments, and new ideas for getting the best from your microscope. Hot off the presses!
CEU's: 0.6 CEUs, 6 P.A.C.E CEU's Microscopy/Microscopy Education adheres to the guidelines established by the IACET.
Pricing: $150 (includes tuition, breaks, course materials, and copy of book) *****Save $25 if paid by 9/22/97 ******* Send three from the same facility and save $50 on tuition for the third person.
Refund policy: Full refund for cancellations made by 10/17/97. After that date, 50% refund or full credit for future class. Substitutions accepted.
Questions: call Barbara Foster or Dr. Ken Piel at MME: (413)746-6931
Registration: Download the form below and fax to (413)746-9311 or call (413)746-6931 and ask for Ken.
Check course you will be attending: ___ Optimizing Light Microscopy (#971003)
Method of Payment: ____ Check enclosed for $ _______________ ____ Visa ____ Mastercard Name on credit card: __________________________________ Credit card number: ___________________________________ Expiration date: ______________________________________ ***If billing address is different from one shown above, please show billing address below: _______________________________________________________ _______________________________________________________ _______________________________________________________
This is a MIME-encapsulated message If you read this, you may want to switch to a better mailer --__==========00000000158236==cellbio.duke.edu==__ Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 8bit
************** Attention all microscopists who might wish to spend a weekend on the coast of North Carolina at THE most beautiful time of the year!!! **********
YOU ARE CORDIALLY INVITED TO PARTICIPATE IN ONE OF THE MORE FUN MEETINGS YOU ARE LIKELY TO ATTEND DURING THE NORMAL COURSE OF EVENTS!
I have attached a textfile for easy download, and also include the complete text as part of this message for those of you who prefer it this way!
Hope to see you in October!
THE NORTH CAROLINA SOCIETY FOR MICROSCOPY AND MICROBEAM ANALYSIS
presents the
SIXTEENTH ANNUAL SYMPOSIUM ON ADVANCES IN MICROSCOPY
"Correlative Microscopy in Biological and Physical Sciences" Blockade Runner Resort, Wrightsville Beach, North Carolina October 17-19, 1997
**************** FOR MORE INFORMATION PLEASE CONTACT PETER INGRAM OR ANN LEFURGEY AT: p.ingram-at-cellbio.duke.edu or a.lefurgey-at-cellbio.duke.edu *************** or READ ON!
Symposium Description The Sixteenth Annual Symposium, sponsored by the North Carolina Society for Microscopy and Microbeam Analysis (NCSMMA), has been planned with a theme of "Correlative Microscopy in Biological and Physical Sciences." Continuing with the tradition of the symposium, the guest lecturers are composed of both nationally and internationally distinguished scientists. The meeting has several purposes, not the least of which is to draw attention of the scientific community to emerging developments in the practical and basic research aspects of exciting new fields, and to bring people together from diverse disciplines to discuss how innovative techniques will be relevant to the future direction of microscopy and microprobe analysis. In particular, this year, special emphasis will be placed on how correlations between the many forms of microscopy are having significant impact in the biological and physical sciences. The symposium also offers an opportunity for interested participants to submit abstracts of related studies for poster display. Three special workshops/tutorials (SEE BELOW) will be offered at no additional charge to participants in the Symposium: (a) Introduction to Scanning Probe Microscopy, ( b) Confocal Microscopy, and (c) Immuno-labeling. These are practical, introductory workshops/tutorials and no previous experience or knowledge is necessary. The annual business meeting of NCSMMA will he held on Saturday, October 18th just prior to lunch.
NCSMMA Student Prizes Student prizes will be awarded at this annual meeting. A total of five awards will be granted to five different students in the following categories: Biological sciences - one abstract (platform presentation) and two posters; Physical sciences - one abstract (platform presentation) and one poster
September 29th is the submission deadline for abstracts of platform and poster presentations. The five winning students will each receive a $100.00 check as well as complimentary registration fees. All abstracts will be pre-judged by NCSMMA members, with platform presentations by the winners, and the posters will be judged at the time of the meeting. Candidates must be members of NCSMMA and be full-time students to be eligible for the competition, and must submit an abstract by the deadline to be considered for either platform or poster awards.
Registration Fees, Hotel rates The $90 ($100 on site) per person and $50 for students ($60 on site) registration fee includes: symposium attendance and materials, Saturday lunch, breaks, and Friday and Saturday evening meals. Additional Friday evening tickets are available for Adults - $20; Children 10 years of age and under - $10. Additional Saturday evening tickets are available for Adults - $20; Children 10 years of age and under - $10. There is a $15 fee for all cancellations. Blockade Runner Resort Hotel special rates start at $69/night including full breakfast.
For questions or further information, please telephone Betty Gooch, Duke University Medical Center: (919) 684-3534 or email: b.gooch-at-cellbio.duke.edu.
PROGRAM
Friday, 17 October, 1997 4:00 - 6:00 pm REGISTRATION and refreshments - Blockade Runner at Wrightsville Beach 6:30 - 8:30 pm Evening Buffet - pool side at the Blockade Runner, Wrightsville Beach Courtesy of JEOL (USA) Inc. and GATAN, Inc. Beverages courtesy of AMRAY Inc. (Complimentary with Registration; Adult and Children Guests $20/$10).
Saturday, 18 October 1997
8:30 - 11:30 am Special Workshops/Tutorials on: (a) Introduction to Scanning Probe Microscopy (hands on participation) (b) Confocal Microscopy (hands on participation) (c) Immunolabeling with Colloidal Gold 9:00 - 11:00 am Coffee, juice, and cookies - Blockade Runner 10:00 - 11:30 am Poster Session, Exhibitors' Displays - Blockade Runner 11:30 - 12:00 Noon NCSMMA Annual Business Meeting 12:00 noon- l:00 pm Lunch - Casual buffet - Poolside
1:00 - 1:15 pm - Welcome -
1:15 - 1:45 pm Correlative Microscopy in Biological Problem Solving Ralph Albrecht 2:00 - 2:30 pm Applications of Scanning Probe Microscopy Chuck Mooney 2:45 - 3:15 pm COFFEE BREAK 3:15 - 3:45 pm Diagnostic Microscopy in the Pharmaceutical Industry Ruth Lightfoot 4:00 - 4:30 pm Electron Backscatter Diffraction in the SEM Joe Michael 4:45 - 5:15 pm Project MICRO: Its Realization and Implementation Caroline Schooley
5:30 - 6:30 pm Poster Session, Exhibitors' Displays 6:30 - 7:00 pm Cocktails, refreshments 7:00 pm Evening Buffet- Al Fresco at the Blockade Runner, Wrightsville Beach Supported in part by Oxford Instruments, Leo and Zeiss
Sunday, 19 October 1997
8:20 - 9:00 am Student Awards and Presentations 9:00 - 9:30 am Correlative Microscopy in Materials Science Mike Kersker 9:45 - 10:15 am Cell Growth Studies with Confocal Microscopy Nina Allen 10:30- 11:00 am COFFEE BREAK 11:00 - 11:30 am EFTEM Techniques in Materials Science Jim Bentley 11:45 - 12:15 Noon Medical Applications of Correlative Microscopy David Howell 12:30 pm Finis
Accommodations Special arrangements have been made to provide a wide variety of accommodations. Use the reservation form immediately and send it directly to the hotel of your choice (except Blockade Runner, which must be telephoned directly) or telephone another hotel directly. Make sure to mention that you are attending the Duke Microscopy Symposium so that you will receive the special rates provided for our registrants.
MAKE YOUR RESERVATION NOW!
A one (l) night's deposit is required to hold reservations. All rates quoted are excluding tax.
Wrightsville Beach, NC (Lumina Avenue) * *BLOCKADE RUNNER RESORT. 1-800-545-5494. Ask for Code #5067. Harbor front $69/single; $77 dbl; ocean front, $89 single/$97 dbl. All rooms include breakfast. * Waterway Lodge (located at drawbridge). 1-800-677-3771. $55/ two dbl. beds or queen size. Condo $65/night. Includes queen and sleeper sofa plus full kitchen.. * Shell Island (at Wrightsville Beach). 1-800-689-6765. Double occupancy suites only. $129/night. * Hampton Inn, 1989 Eastwood Road. 1-919-0256-9600. $89/king or two queen beds plus includes continental breakfast with local calls. * Meeting site
Wilmington, NC (Market Street) * Greentree Inn, 1-910-799-6001. $43.95//two dbl. beds w/ continental breakfast. * Holiday Inn of Wilmington, 1-910-799-1440. $75/king or two double beds. * Howard Johnson Plaza, 1-800-833-4721. 89/night, two double beds or one king size bed. * Days Inn, 1-910-799-6300. $58.88//two dbl. beds. or 1 queen
************* Send checks payable to: "Sixteenth Annual Symposium on Advances in Microscopy" Send to: Betty P. Gooch, Symposium Coordinator Analytical Electron Microscopy Facility Box 3709 Duke University Medical Center Tel: (919) 684-3534 Durham, NC 27710 email: bgooch-at-cellbio.duke.edu Fax: (919) 681-8419
PLUS!!!!!
3 SPECIAL FREE WORKSHOPS!
TO BE HELD IN CONJUNCTION WITH THE
SIXTEENTH ANNUAL SYMPOSIUM ON ADVANCES IN MICROSCOPY
Sponsored by the North Carolina Society for Microscopy and Microbeam Analysis at the Blockade Runner Beach Resort Wrightsville Beach, North Carolina October 17-19, 1997
INTRODUCTION TO SCANNING PROBE MICROSCOPY: Application to Materials Science & Biology Chuck Mooney, Park Scientific Instruments, Sunnyvale, Ca.
Basics of STM and AFM
* Contact, non-contact and tapping microscopy * Problems and solutions * Lateral force microscopy * Operating parameters ***** Hands on participation ******
CONFOCAL MICROSCOPY Nina Allen, North Carolina State University, Raleigh, NC
Basic Operating Principles
* Problems, solutions and limitations * Laser sources * Confocal versus conventional fluorescence imaging * Slit and pinhole apertures and deconvolution * Fluorescence throughput * 3-D imaging * Alignment of optics * Judicious use of dyes * Sensitivity and background subtraction
***** Hands on participation ******
IMMUNOLABELING WITH COLLOIDAL GOLD Ralph Albrecht, University of Wisconsin, Madison, WI
* Production of gold colloids * Conjugation of antibody/gold ligand to colloids * Basics of labelling with colloidal gold conjugates * Techniques for silver enhancement of gold colloids * Problems, solutions and limitations
Use of gold conjugates as labels in correlative microscopy
We are interested in buying a used cryo-ultramicrotome. LKB, Reichert, or RMC acceptable. Please contact: Marek Malecki, PI Phone: 6082638481. Fax: 6082654076. Email: malecki-at-macc.wisc.edu
I was recently given the following homework assignment: Actual resolution as determined by measuring the smallest distance between two distinguishable points in a suitable specimen (resolution standard), is usually greater than the linear resolution of the microscope. Describe the factors that contribute to this discrepency. Find and list suitable good references. Can you give me any help in solving this problem? Do you know of any good sources to speak to? Do you know the answer? Thank you for your help, Fred Meisenkothen
I think that there must be somebody who have got experience with EDS for Al-alloys. I hope you can shine some lights on my silly questions.
We have several students working on Al-alloys (e.g. Al-Fe-Ti, Al-Fe-Cr et al) using arc melting. The problem is that the EDS results showed that all the alloys (as-cast or following solution annealing) lost about 20at% Al (for example, {20% for the nominal composition 25%)?! We tried on two machines, An-10000 EDS on JEOL 840 (over 10 years old) and Oxford ISIS EDS on FESEM Hitachi S-4500 (only 1 year old). They showed similar results at work conditions of 20kV, } 1000cps, for 100s on well polished samples.
Of course, there are several posibilities for the large amount of Al loss. Weight ratios have been double checked. Melting under Argon would cause little loss of Al because the melting point of Al is reletively lower than others, and some dark dust did appear in the furnace, but this error should be limited in +/-0.5%. Where did the Al go?
So, I wonder if there is anything wrong with the standardless analysis of EDS. We have tested the EDS with stanless steel sample. The results were well consistant with the nominal compositions. Is that because there are something wrong with the standard data of Al in the computer, or calibration, or work conditions, or something else? Any idea? Please help.
} Could I suggest the original poster try the Safety group. The address is } } LISTSERV-at-UVMVM.UVM.EDU } Write SUB SAFETY in the text area.
Good suggestion. There was a discussion within the last few years on this topic on the safety listserv. The folks at the University of Vermont who run the list also have an excellent website at: {http://siri.org} where they archive messages from the listserv, as well as maintain access to MSDS information and other safety related information. Full instructions for subscribing to the list are also there.
Among mostly serious information it also contains the following spoof of the Good Times virus hoax that is just too good not to repost here, although it of course has nothing whatsomuchever to do with translation. Same could be said about Join the crew, I believe...
READ THIS:
Goodtimes will re-write your hard drive. Not only that, but it will scramble any disks that are even close to your computer. It will recalibrate your refrigerator's coolness setting so all your ice cream goes melty. It will demagnetize the strips on all your credit cards, screw up the tracking on your television and use subspace field harmonics to scratch any CD's you try to play.
It will give your ex-girlfriend your new phone number. It will mix Kool-aid into your fishtank. It will drink all your beer and leave its socks out on the coffee table when there's company coming over. It will put a dead kitten in the back pocket of your good suit pants and hide your car keys when you are late for work.
Goodtimes will make you fall in love with a penguin. It will give you nightmares about circus midgets. It will pour sugar in your gas tank and shave off both your eyebrows while dating your girlfriend behind your back and billing the dinner and hotel room to your Discover card.
It will seduce your grandmother. It does not matter if she is dead, such is the power of Goodtimes, it reaches out beyond the grave to sully those things we hold most dear.
It moves your car randomly around parking lots so you can't find it. It will kick your dog. It will leave libidinous messages on your boss's voice mail in your voice! It is insidious and subtle. It is dangerous and terrifying to behold. It is also a rather interesting shade of mauve.
Goodtimes will give you Dutch Elm disease. It will leave the toilet seat up. It will make a batch of Methanphedime in your bathtub and then leave bacon cooking on the stove while it goes out to chase gradeschoolers with your new snowblower.
Listen to me. Goodtimes does not exist.
It cannot do anything to you. But I can. I am sending this message to everyone in the world. Tell your friends, tell your family. If anyone else sends me another E-mail about this fake Goodtimes Virus, I will turn hating them into a religion. I will do things to them that would make a horsehead in your bed look like Easter Sunday brunch.
Colleagues, I am wondering how one would keep the particles from falling through the open holes since the particles in general are smaller than the size of any holes in a holey film. Also, how do your "Ultra-thin carbon films" differ from the ones I purchase from Agar Scientific in the UK and from SPI in the USA? Are they really "thinner" on the basis of some measurement and if so, what measurement do you use?
Sincerely
.attila(L)
Attila L. Toth ------------------------------------------ MTA MFKI Research Institute for Technical Physics of the Hungarian Academy of Sciences H-1325 Budapest POB 76 tel: (36.1) 169-2100 x 226 fax:(36.1) 169-8037 ------------------------------------------ email: tothal-at-mufi.hu (EUDORA:=CDrj =E9kesen!) ------------------------------------------
Really need a bit more information, but... Have you examined the material for homogeniety? BSE imaging of a polished surface would be a good start. If you have more than one phase / precipitates, apparent composition will be a strong function of where you analyze and the size of the analysis volume relative to the phases/inclusions. Generally I have found that inhomogenous materials will compromise analysis accuracy.
Woody White, Electron Microscopist SEM/EDS/WDS
Work: Mcdermott Technology, Inc. woody.n.white-at-mcdermott.com http://www.mtiresearch.com/
Dear All I was experimenting with new mail filtering and inadvertenetly sent mail to a number of addressess that were not supposed to be mailed. Many apologies if you receive that mail either directly or via the list
Chris
Chris Gilpin Biological Sciences Electron Microscope Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 161 275 5170 fax +44 161 275 5171 http://www.biomed.man.ac.uk/biology/emunit/emhome.html
} Here is what I propose: take some epoxy resin and coat the inside of a } polypropylene beaker with the epoxy by building up layers and polymerizing } the layers.
} Allow the uranium liquids to evaporate } to dryness (taking care to avoid generating dust) and apply another layer } of epoxy over the uranium salts.
I have to emphasize the point about avoiding dust. Uranium is an alpha-emitter (not a beta-emitter as another poster said, although some of the daughter nuclei emit betas), and the range of these alphas is so short that they will not penetrate the dead layer of the skin. Thus, uran- ium is not dangerous unless it is ingested or inhaled. If, however, uran- ium is inhaled, particles sitting on the lung cells will provide a very large dose to those cells, and are a serious carcinogen. It is much safer for everyone if there is NO chance of producing dust or droplets containing uranium; I'd even be very careful about pouring the liquid into a beaker. Yours, Bill Tivol
Attila L. Toth wrote: ================================================== I am wondering how one would keep the particles from falling through the open holes since the particles in general are smaller than the size of any holes in a holey film. Also, how do your "Ultra-thin carbon films" differ from the ones I purchase from Agar Scientific in the UK and from SPI in the USA? Are they really "thinner" on the basis of some measurement and if so, what measurement do you use? ================================================= You are right, CdS nano- particles, at least the ones we have seen, those that would be thin enough to "see" through, are smaller than the smallest holes we have ever seen anyone able to make in a "lacy" film. So I think that there might have been some misinformation accidently posted about this a while back (see Aug. 11 posting DUNNTEM-at-aol.com working for Ted Pella, Inc .). If I am wrong about this please correct me and set the record straight .
With regard to so-called "ultra-thin carbon films", obviously carbon films can be made almost infinitely thin, but they do have to have enough mass to be self-supporting on the grid mesh being used. That minimum amount of mass then is going to be a function of the mesh size, the lower the mesh size (e. g. larger the hole), the more durable (a.k.a. thicker) must be the carbon film. If thickness of the carbon film is crucial, e.g. you want it completely minimized, I would recommend our finest, which is our 2000 mesh grid. This simple reality is often times missed by people worried about film thickness (which in fact ought to not even be relavent in the case of a lacey or holely film, because after all, you are getting your information through the holes, not through the "lacy" areas). So the whole need to worry about "thickness" per se really ought to be, in the case of lacey films, a non-issue!
But addressing the subject of carbon film thickness generally, when one does have the need to "look through" the film, the philosophy of "one thickness fits all" is not one to which we subscribe. One can have the luxury of having that philosophy only if there are in-house facilities available to quality check what has been made, otherwise the customer ends up doing the quality checking. And finding out at the last minute that coated grids are not stable and can not be used is usually not a very pleasant discovery.
So while I can not comment on the methods used by our competitors to measure film thickness, I can tell you how we do the "test" and that is, as we are making them, and we strive for the minimum possible amount, before we have made too many, we walk across the hall and put them into a TEM that is dedicated (after 5:00 pm) for this purpose, that is, to do our own "clinical" test to make sure that the film is sufficiently durable (e.g. thick enough) for that given mesh size being used. Now you might not believe this, but the instrument used for the testing is an RCA EMU 4-B, manufacturerd by RCA in 1969. So we think of this as a worst case test. If the carbon film is stable enough to survive the beam of an RCA EMU-4B, an instrument featuring technology more than thirty years old, surely it ought to be more than acceptable in an instrument of more modern manufacture (assuming minimum column cleanliness). And that kind of test procedure results in a history of almost (note I said "almost") no returns!
Interestingly enough, the films do not have to be "thicker" to be stable in the RCA than in our much newer JEOL TEM. A film with minimum thickness, stable in one, seems to be similarly stable in the other.
More information about our custom coated carbon grids can be found on our website given below.
Disclaimer: SPI has produced custom coated grids for customers for more than twenty years so we have an interest in promoting our carbon coated grids. The posting I characterized as containing "misinformation" came from a competitor, so everything I said should be taken within that context.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Jim Darley wrote: ================================================= But too many regulations are not wise - eg. the dangerous goods shipping laws which appear designed to make things more expensive but not safer. ================================================= We should all be taking the HAZMAT reguations, no matter where we live, seriously. I can not comment about the laws in Australia, but I have a very high regard for the regulations promulgated by the U. S. Dept. of Transportation (DOT) and IATA (for international shipments) as well as the reasons behind them. And I can state, from first hand experience, that if the regulators are presented with sound technical information for a reguation to be changed, they will indeed listen (at least in the US) and sometimes make changes.
If anyone remains unconvinced, about the importance of adherence to all HAZMAT regulations, keep in mind that the ValuJet disaster was caused by someone not taking seriously these same regulations.
While it is correct that one can incur almost unbelievalbly high shipping costs for HAZMATs, this is not always the case. In many instances, such as for the ordering of osmium tetroxide, if one orders "smart", they can do their shipping at costs only nominally more than if the same weight of tweezers, grids, or SEM mounts were being shipped. Now this would not apply for everything (e.g. our SPI Dusters, for example) but it does apply for a surprising number of HAZMATs routinely used in EM labs. We are also in the process of reformulating some of our embedding kits so they too can be shipped at the lower prices.
We have tried to explain how a customer can "order smart" on our website, click on "Hazardous Items(Good News and Bad News)". While savings in shipping costs for domestic US customers are possible, the real beneficiaries are foreign customers now no longer are restricted to the use of air freight (which has associated with it high minimum charges).
And while we are on this subject, just remember, don't ever try to take HAZMATs in checked airline luggage to save some money, it is unconscienable from a moral standpoint and puts at risk everyone on the plane flight.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Jim Darley wrote: ================================================= But too many regulations are not wise - eg. the dangerous goods shipping laws which appear designed to make things more expensive but not safer. ================================================= We should all be taking the HAZMAT reguations, no matter where we live, seriously. I can not comment about the laws in Australia, but I have a very high regard for the regulations promulgated by the U. S. Dept. of Transportation (DOT) and IATA (for international shipments) as well as the reasons behind them. And I can state, from first hand experience, that if the regulators are presented with sound technical information for a reguation to be changed, they will indeed listen (at least in the US) and sometimes make changes.
If anyone remains unconvinced, about the importance of adherence to all HAZMAT regulations, keep in mind that the ValuJet disaster was caused by someone not taking seriously these same regulations.
While it is correct that one can incur almost unbelievalbly high shipping costs for HAZMATs, this is not always the case. In many instances, such as for the ordering of osmium tetroxide, if one orders "smart", they can do their shipping at costs only nominally more than if the same weight of tweezers, grids, or SEM mounts were being shipped. Now this would not apply for everything (e.g. our SPI Dusters, for example) but it does apply for a surprising number of HAZMATs routinely used in EM labs. We are also in the process of reformulating some of our embedding kits so they too can be shipped at the lower prices.
We have tried to explain how a customer can "order smart" on our website, click on "Hazardous Items(Good News and Bad News)". While savings in shipping costs for domestic US customers are possible, the real beneficiaries are foreign customers now no longer are restricted to the use of air freight (which has associated with it high minimum charges).
And while we are on this subject, just remember, don't ever try to take HAZMATs in checked airline luggage to save some money, it is unconscienable from a moral standpoint and puts at risk everyone on the plane flight.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
With regard to references the following are quite good for sizing in the optical microscope:
Handbook of Chemical Microscopy by Chamot and Mason
The Particle Atlas, Volume 1 by McCrone and Delly
Particle Size Measurement Vol 1 by Terence Allen
I have some trouble understanding the question, it doesn't seem to address any issue precisely since it does not specify the optics, the resolution standard, the wavelength of light used or the method of measurement. All those factors affect the resolution. Real measurements are also affected by the contrast in the specimen, the calibration standard, and such mundane issues as how well the optics are aligned and adjusted (ie the use of Kohler illumination, etc.) and how young and fit the eyeballs are doing the measurements - even the lighting in the room can affect measurements.
The questioner may be after how diffraction affects particle measurements. The limit of detection of the optical microscope is much less than the resolution limit. I can detect objects much smaller than 0.4 um (the resolution limit of my neofluor 40 times objective with white light) - probably down to 0.1 um, but diffraction effects make them appear larger so that they all look to be about 0.4 um. Allen's book has a nice description of this effect and the other two books have good discussions of the origins of the resolution equation.
Hi- I am forwarding this message for a colleague who asked me this question. I don't have a clue. Any help appreciated. Dave
} The calibration of the cell for pH measurements using fluorescent probes } is done by using ionophores such as nigericin or by ratiometric calibration. } Does any of this method account for the effect of the dielectric constant } of t } the medium on the pKa' of the probe ? If not is the effect of the } dielectric co } nstant of the medium accounted by any other method ? Is it justified to } ignore the effect of the dielectric constant of the medium ? } } Thank you in advance for your help and would be eagerly waiting for the } respo } onses. } Abizer Harianawala }
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut U-125 Storrs, CT 06269 Knecht-at-uconnvm.uconn.edu
Dr. Jarett, The following ad has been posted on the Microscopy listserver which exclusively serves the em community. I did not include the email address on purpose to prevent unnecessary postings in your mail box. Hope you have a pleasant weekend. Neelima
ELectron Microscopist:
The University of Pennsylvannia School of Medicine Institutional Electron Microscopy Core is searching for a Co-Director. This Core has the broad support of the Diabetes Center, the Cancer Center, the Department of Pathology and Laboratory Medicine, and School of Medicine. The Core is widely used by medical community, as well as the University as a whole. The candidate should have a Ph.D. in cell and molecular biology and at least 5 years experience with all aspects of Electron Microscopy. The individual should also possess administrative experience and computer skills. The position title will be Senior Research Investigator and will be responsible for helping the Director develop reports for centers, grants, papers etc. Salary will be consistent with experience. Respondents should send their curriculum vitae, bibliography, and 3 references to: Dr. Leonard Jarett, M.D. Chair, Department Of Pathology and Laboratory Medicine Universtiy Of Pennsylvania 6 Gates Building 3400 Spruce Street, Philadelphia, Pa 19104-4283 Regards... : ) ; ) Visit us at http://www.med.upenn.edu/~path/core/EMCMAIN1.HTM
You should always be careful regarding ZAF corrections for mixtures of light and heavy elements. My advice is, if you don´t have a suitable standard to test your ZAF corrections, then don´t trust them. You have not calibrated your ZAF corrections until you test them on a suitable standard. For mixtures of Al-Fe-Cr, use a similar standard, not just Fe-Cr or Fe standards.
Sorry Bill but Uranium emits a whole zoo of radiations, alpha, beta, gamma. A jar of UA placed on a sheet of fast film for a day or two will expose the film through the jar. UA dissolves beautifully in water and the sitting in the lung argument applies more to other forms of U - like in mining. I maintain that poring small quantities (I used to discard 5ml/months of a 2% solution) into the sewage system is the most sensible solution. Just sit down and work out the dilution factor on an annual basis and I expect in most cities the result will be approximately that concentration of U in seawater. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 400+ Links, MSDS ************************ http://www.proscitech.com.au
---------- } From: William Tivol {tivol-at-wadsworth.org}
} } Dear John & others, } } } Here is what I propose: take some epoxy resin and coat the inside of a } } polypropylene beaker with the epoxy by building up layers and polymerizing } } the layers. } } } Allow the uranium liquids to evaporate } } to dryness (taking care to avoid generating dust) and apply another layer } } of epoxy over the uranium salts. } } I have to emphasize the point about avoiding dust. Uranium is an } alpha-emitter (not a beta-emitter as another poster said, although some } of the daughter nuclei emit betas), and the range of these alphas is so } short that they will not penetrate the dead layer of the skin. Thus, uran- } ium is not dangerous unless it is ingested or inhaled. If, however, uran- } ium is inhaled, particles sitting on the lung cells will provide a very } large dose to those cells, and are a serious carcinogen. It is much safer } for everyone if there is NO chance of producing dust or droplets containing } uranium; I'd even be very careful about pouring the liquid into a beaker. } Yours, } Bill Tivol }
I would guess that you are trying to push standardless EDS a bit further than it's intended to go. Why don't you use some standards?
Ritchie
} We have several students working on Al-alloys (e.g. Al-Fe-Ti, } Al-Fe-Cr et al) using arc melting. The problem is that the EDS results } showed that all the alloys (as-cast or following solution annealing) lost } about 20at% Al (for example, {20% for the nominal composition 25%)?! } Where did the Al go? } So, I wonder if there is anything wrong with the standardless } analysis of EDS. } Charlie Kong
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
We are continuing to improve the US Histochemistry Society Web page and welcome your suggestions as to how we can be of better service to the community.
Right now, we are running a link to the Immunocytochemistry Discussion Newsgroup. Check out our page for instructions on how to get involved. It is: http://www.hcs.microscopy.com
We are also running a contest for the best Logo for our society. We have ten entries and welcome more. The prize is $200.00. Check out the Logo contest on the above web site and vote for the best and submit your own.
Thanks for your attention!
Gwen Childs
************* Gwen V. Childs, Ph.D. Professor and Vice-Chair Department of Anatomy and Neurosciences University of Texas Medical Branch Galveston TX 77551-1043 gvchilds-at-utmb.edu http://cellbio.utmb.edu/childs/childs.htm (409) 772-1942; FAX 772-3222 Toll Free Pager: 1 888 715-8636
I have an old EDAX 9100 detector with broken Be window and photo diode plus FET. I like to know whether (1) someone could fix it quickly with affordable price or (2) some information on where we could buy the Be window and the diode-FET unit, plus tips to fix the detector. My students and I would appreciate any help from you and we hope we could have our detector back to work so we could continue our experiments.
Thanks so much,
Judy Wu Dept. of Physics Univ. of Kansas Lawrence, KS 66045 (785)864-3240 (phone) (785)864-5262(fax)
/////////////////////////////////////////////////////////////////////////////// This Message was Composed using Extractor Pro Bulk E- Mail Software. If you wish to be removed from this advertiser's future mailings, please reply with the subject "Remove" and this software will automatically block you from their future mailings. "Remove" must be in the "Subject Field". //////////////////////////////////////////////////////////////////////////////// Attn All Businesses : Increase Profits Immediately ____________________________________________________
If you operate a business or in the process of starting one, please
pay close attention to this notice. You will increase your profits by
increasing your access to new customers, increasing your
receivables, increasing your sales, and saving you money on
services you may already have. Our offer is GIMMICK-FREE so
please read the entire notice so that you may determine whether
or not our offer can benefit your company. Our offer is Good
for new businesses or established businesses. Owners with good credit
or unfavorable. Internet, mail or phone, work-at-home, or retail
establishments qualify. For less than $40 per month your business
will be furnished with :
A Merchant Bank Card account-with no monthly statement fee and a 1.39% rate for retail-swipe transactions, and a low 2.39% rate for non-swipe (mail/phone orders). Both carry a 20 cent per transaction fee. Comes with your own software to install into your own computer, or, comes with your own separate terminal.
You will increase business immediately, even if you believed you could not qualify, or thought it was to expensive, or felt it was to complicated. You will begin excepting ALL major credit cards immediately, using a bank card terminal, or your own computer if you prefer. Even if you are in a high-risk industry (computer mail order, gambling, telemarketing, travel agency and health clubs). All business invited, especially work-at-home and internet merchants, and even flea-market merchants... even if new in business.
HOW IS THIS DIFFERENT FROM OTHER OFFERS ? Please read below carefully.
First, there is no risk to the applicant, since there is no application fee. Next, we have no monthly statement fees, common to 99% of the banks in the industry. Next, we have really low processing rates, including a phenomenal 1.39% rate for retail accounts. For mail order, our per-transaction fees are a phenomenal 20 cents, not 30 cents common to the industry. But this is what separates us from the competition. Obtain your merchant account today from our company, and you will receive the following services free!
1. Accept checks by fax or e-mail-immediately increases your sales and collections. Accept business or personal checks by phone, deposit them into your bank account the very same day! ($310 value)
2. 800 / 888 services set up at 9.4 cents per minute. Increase your business by giving your perspective customers easy access to you. No monthly fees, no setup charges, excellent rates, guaranteed 24hrs a day, seven days a week with incremental billing! ($180 yearly value)
3. Get new customers-free! Our e-mail software will allow you to literally send out to millions of prospects your business offer. Our competition is selling this software for $1000 and more. It is one of the best kept secrets in the industry, and is already revolutionizing the way many business are approaching there direct-mail campaigns... by practically eliminating them and using e-mail to reach more prospects with out the cost of postage and materials! ($999 value)
4. FREE WEB PAGE (2 pages + an online order form = 3 pages) setup. Not only that, we will host and list your "WEB STORE" in over 450 search engines for three months-FREE. Again, free web page setup, and free web page listing and hosting, for 3 months. ($250 value)
For less than $40 per month, you will receive a merchant account for accepting all major credit cards, which INCLUDES your processing Terminal or software. You will receive checks by fax, You will receive a free 800# with no monthly fees, You will receive our e-mail software to attract new customers, and you will be setup with your own web page and have it hosted and listed for 3 months, free of charge. Whatever it is you are selling, weather it is to increase sales or get started, YOU WILL NEVER come across an offer such as this one, offering so much, for so little to start. Better yet, there is no risk to get started, since there is no application fee for your merchant account (unless you are a high-risk merchant, than a refundable application fee would apply). We are giving you every reason to consider our offer and get started immediately. In fact, we also offer an incentive for those merchants who get started right away- FREE CHECK COLLECTION (avoid the hassles of bounced checks).
For more information please hit the REPLY BUTTON and type 400AD in either the subject line or body of the e-mail. (Please include your phone number). You will receive further information via phone or e-mail and a preliminary application designed to assist us determine the best bank for your merchant account.
Or call us directly: (813) 508-2676 during business hours, or fax us 24 hours (813) 532-6996) - please use reference # 400AD in any communication to our company in order to qualify your web page service, e-mail software, 800# setup, and checks by phone/fax system that come with your Visa/MC/Amex merchant account.
How to contact us in writing :
Electronic Processing of NY/financial services, 520 third ave, suite 310, New York, NY. 10016.
Financial services and solutions works with major companies enabling us to bring you the best services available in America without having to pay expensive middle-man fees. Our services have the following in common : Increasing your sales, increasing your collection efforts, increasing your customer traffic, and saving you money. We offer ATM services (placing a physical ATM in your establishment), collection services, check collection software, check guarantee services, fax-on-demand services, ACH wire transfer services, internet web services, and telecommunication services to name a few. Contact us today and make us your one-stop shopping center for all your business needs.
NOTE : Only owners with undischarged bankruptcies (currently unresolved) will automatically declined from our merchant account (Visa/MC/Amex). They can still apply for all our other services, still priced at competitive, wholesale pricing.
This is a multi-part message in MIME format. --------------D0A6E5E30BCDAEC778B68638 Content-Type: text/plain; charset=iso-8859-1 Content-Type: text/plain; charset=iso-8859-1 Content-Transfer-Encoding: quoted-printable X-MIME-Autoconverted: from 8bit to quoted-printable by iris.microcosm.com id SAA03611
Dear Microscopists:
We would like your help in locating the best candidate for the position of Director for our Applications Laboratory. The following text and attachment describe the opportunity. Would you please post the advertisement and bring it to the attention colleagues?
IMAGING RESEARCH OPPORTUNITY
We are seeking a talented individual to direct our Applications Laboratory, leading research into fluorescence detection, confocal infrared and ultraviolet microscopy, and x-ray microscopy. The incumbent will also participate in the specification and design of novel optical systems developed by the Engineering Department. The Applications Laboratory patents and publishes research findings, supports Microcosm=92s customers=92 in their use of our technology, and communicates the implications of internal and external research to the Marketing and Engineering Departments.
The position requires a Ph.D. in cell biology or a related science, with primary experience in microscopy and imaging, and a strong physical science background. Familiarity with the latest microscopy techniques is mandatory. It is essential that the incumbent has sound knowledge of optics and digital image analysis, both in theory and in practice. Ideally candidates will have experience with several imaging platforms, including isee and dsp/os, NIH-Image, Image Tool, Metamorph & Image-1, as well as the LSM and MPM instruments made by Carl Zeiss.
The compensation package for the right candidate is negotiable. Benefits will include medical and dental insurance, and participation in our profit-sharing 401(k) retirement plan.
Interested persons should send their r=E9sum=E9 to the attention of Dr. Patrick Huddie at the address above. Please direct e-mail to phuddie-at-microcosm.com.
-------------------------------------------------------------- Dr. Patrick L. Huddie (301) 725-2775 Fax (301) 725-2941 Microcosm, Inc., 9140 Guilford Road, Suite O, Columbia, MD 21046 e-mail phuddie-at-microcosm.com URL http://www.microcosm.com The Web is:"Vaster than empires and more slow" Andrew Marvell (1621-1678)
--------------D0A6E5E30BCDAEC778B68638 Content-Type: text/x-vcard; charset=us-ascii; name="vcard.vcf" Content-Description: Card for Patrick Huddie Content-Disposition: attachment; filename="vcard.vcf" Content-Transfer-Encoding: 7bit
begin: vcard fn: Patrick Huddie n: Huddie;Patrick org: Microcosm, Inc. email;internet: phuddie-at-microcosm.com title: CEO x-mozilla-cpt: ;0 x-mozilla-html: TRUE end: vcard
A good place to start is "The Image Processing Handbook" Second Edition. John C. Russ. CRC Press. ISBN 0-8493-2516-1
Mel Dickson President, Australian Society for Electron Microscopy Director, Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
Hi, Thank you who kindly responded to my question on EDS for Al-alloys. Troublesome students always raise extraordinary questions to test the range of knowledge (and/or self-confidence) of their teachers. Sometimes, they even try to meltdown the reputation of modern technique with the spark of their intuition. Now, let's show our royalty to the advanced materials science. The big question mark has not been erased yet, although the simplest answer, which may be the right one and has been selected, is to blame the students who prepared the dummy samples. Who knows whether they swallowed a piece of aluminium down before melting or not? There were some beginners who asked me quite often about the error range and sensitivity of EDS. My answer is that at first it depends upon your sample (Am I a good lawyer?); secondly, in micro-scale few thing is uniform (Do not blame me if the results are unrepeatable); at last, I would say that in ideal testing conditions the error should be less than +/-0.5% in general(I hope so indeed!). They were quite happy to use the machine. Several experts have advised me that we should give up the dependence upon the standardless analysis of EDS, although the others showed strong evidence to prove that it is working well. Faster would never be safer, just like driving a car. This should not imply that the results of "semi-quantitative analysis" can only be trusted in HALF, neither the error may be 50%. How do you feel? Do you mention "semi-" quite often to the users? My former colleague, Bruce, suggested that oily detector window might cause the problem for softer radiation such as Al-Ka. This message struck a spark in my poor memory. I really saw somebody who dared to wipe oil off the thin Be-window of a TN-5400 EDS using a cotton ball with a drop of acetone. That was ten years ago. Let's go to the question: Have you ever cleaned the Be window of your EDS system? If the answer is positive, how often? Or, just wake me up ---- NEVER EVER THINK ABOUT THAT, YOU THE TROUBLE MAKER! I swear I have never been a trouble-maker. I just want to learn something from you. Just talking, no action following. It is too lang. Thank you for your time.
A professorship due to the retirement of Professor Jon Gj=F8nnes at University of Oslo has been announced. The professor is expected to strengthen the reseach activity of the department in materials science/structure physics. The activity is mainly concerned with the application of electron-optical techniques to metallurgical questions, ceramic and semiconductors and with the development of quantitative methods for obtaining structural information (crystal structure, electronic structure and disordered structure).
Here follows the official announcement, including the application procedure. The announcement can also be found in the Journal Nature. The application deadline is October 3 1997. For additional information please contact
Professor Arne Olsen Department of Physics/Centre for Materials Science Gaustadalleen 21 0371 Oslo Norway Tel. (+47) 22 95 87 40 Fax: (+47) 22 95 87 49 e-mail: arne.olsen-at-fys.uio.no
Professor in Physics (Materials science/Structure physics)
The Faculty of Mathematics and Natural Sciences at the University of Oslo invites applications for the position of Professor in the Department of Physics with research interests within the field of materials science/structure physics.
The Department of Physics at the University of Oslo has 83.5 academic staff of which 26 are temporary (17 research associates and 9 adjunct professor positions). Further,there are about 45 funded by external sources. The department has 38 technical staff positions and 9 administrative staff positions.
Teaching in the Department is directed towards the degrees cand. mag. (approx. B.Sc.), siv.ing. (M.Eng), cand.scient. (M.Sc.) and dr.scient. (Ph.D). There are currently 110 students enrolled in Masters programmes and 70 in doctoral programmes.
Research in the department is organised in 8 groups which undertake both experimental and theoretical studies: Biophysics, Electronics, Elementary Particle Physics, Condensed Matter Physics, Nuclear and Energy Physics, Plasma and Space Physics and Structure Physics. In addition there is a Theoretical Physics Group.=20
The vacant professorship is connected to the Structure Physics Group, which also is part of the Faculty's Centre for Materials Science. The Structure Physics Group, localized in the Oslo University Research Park, runs an electron microscopy and metallographic laboratory with two transmission electron microscopes (JEOL 200CX and 2000FX) equipped for analysis (EDS, EELS, TV-system), optical microscopes and image analysis. A new TEM instrument will be installed in 1998. The research group, which numbers 20-25 students and staff, has extensive collaboration with other research groups within the University and in Norwegian industrial and public research laboratories. The group also maintains strong international contacts. The academic staff are engaged in the University's teaching programme for materials science as well as in general physics teaching in the department.=20
Much of the research in the group is linked to the application of electron-optical techniques to metallurgical questions, ceramics and semiconductors and to the development of quantitative methods for obtaining structural information (crystal structure, microstructure, electronic structure and disordered structure).
The professor will be expected to strengthen the research activity in Structure Physics as well as the operation of the group's laboratories. The successful applicant must be able to document scientific expertise in one or more of the group's major activities.=20
The appointee must be able to provide supervision at all levels of teaching. The professor will have responsibility for supervision of masters and doctoral candidates within her/his special field. The language of instruction for undergraduate courses is Norwegian, but English will be accepted for the first three years of appointment. The appointee may also be required to undertake administrative duties as prescribed in the applicable University regulations.
According to current regulations, the evaluation of applicants takes regard of scientific, professional and educational qualifications as well as other activities, which may qualify the applicant. Where several applicants are deemed to have equivalent qualifications after evaluation of the scientific, professional and educational qualifications, a female applicant will be ranked above a male applicant according to the procedure for the appointment of professorial staff.
The application shall specify the candidate's education, previous positions, scientific, professional and educational activities and administrative experience. Curriculum vitae and publications list are therefore to be included.
The application shall furthermore include a short description of the scientific works that the applicant regards as the most significant and on which the evaluation might especially be based. Normally this ought not exceed 10 items.
The application is to be addressed to the Academic Collegium, University of Oslo, and is to be sent with documentation to: Faculty of Mathematics and Natural Sciences, P.b. 1032 Blindern, N-0315 Oslo, Norway by the application date. Within one month of this date, the applicant must have sent to the Faculty's Secretariat:
- 5 complete sets of scientific works, published or unpublished, which one wishes to be considered in the evaluation (normally not exceeding 10)
- 5 copies of the application with documentation (C.V., complete publications list, description of the 10 most important works)
Scientific works which are in preparation on the application date may nonetheless be submitted within three months of the deadline provided the Secretariat is informed when the remaining works are submitted.
After the application date, the University will send applicants instructions for submission of scientific works.
One is otherwise referred to the regulations for appointment of professorial staff approved by the Academic Collegium in accordance with the University Act =A732.
I thank Garratt-Reed for corroberating my previous posting. John Bazzola had asked for a rough guide (he has confirmed that since) on detection limits of EDS versus WDS techniques. Anybody who has any significant experience with microprobe analysis knows that there is no single line correct answer. However, it is imperative for analysts to remember a few general figures so they can advise on appropriate instrumentation and techniques.
John B's initial inquiry deserved a reply and when none was given, I posted mine more than a day later. I believe that my posting is a useful guide for non-specialist analysts. Nothing that GR writes makes nonsence of my posting.
Nobody had asked about other differences between the techniques eg. resolution or simultaneous acquisition. I am pleased that G-R has supplied some information on those topics and on new, very high count-rate acquisition facilities for EDS. G-R is proud of his 0.6% "quantitative analysis" of Cr at an accuracy of +/- 0.1wt%.
I suggested that in EDS the presence of 1% of an element is the approxiamte lower limit for quantitative analysis. G-R has lowered that limit by some 40% - or has he? +/- 0.1% is good when 20% of the element is present, as it represents an accuracy of 0.5%. +/- 0.1% when 0.6% is present is about +/- 16% accuracy; I call that qualitative or at best semi-quantitative.
Another correspondent emailed me and noted that it was President Trueman who had been looking for a one handed adviser. But that was for an economist and not a scientist as I, apparently wrongly, remembered. The correspondent could see my point though; thanks to Brian Demczyk. I think that Trueman had an excellent idea but he should have extended that search to a scientist as well. Certainly microscopists and economists share disciplines which combine art and science. And I should add require 'good judgement'.
Why now was I abused with that opening: "Jim Darley's posting is hardly furthering good science". Am I to believe that good science is advanced by quarelsome nitpicking and that all broadbanding is verboten? I plead not guilty. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 400+ Links, MSDS ************************ http://www.proscitech.com.au
} } Subject: Re: detection limits x-ray } } Date: Friday, 5 September 1997 6:21 } } } } Jim Darley's posting is hardly furthering good science. } } } } 1: Detection limits (John's original question). Modern EDX systems are } } easily capable of quantitative analysis in the sub-1wt% range. See, for } } example http://prism.mit.edu/facltis/stem/stmexam.htm (the second } } illustration on that page) where I was getting analyses for Cr in steel } of } } the order of 0.6+/-0.1 wt%. I don't know about you, but I consider this } } quantitative. This, mind you, was in a measurement where I was } attempting } } to optimize spatial resolution rather than sensitivity, and the } acquisition } } time was 60sec. per data point. This example, of course, was from a } STEM. } } There are many more examples in the literature. } } } } Using beam gating techniques such as those developed by Charlie Lyman and } } colleagues, EDX data can readily be acquired at 20,000-40,000 counts per } } real second, if spatial resolution is sacrificed. Combine this with an } } acquisition time of 1,000 seconds (by no means unrealistic for an } important } } measurement) and the detection limit is in the rage of, or better than, } 0.01wt%. } } } } Of course, in the SEM, which may have been the point of John's original } } question, the situation is not the same, the beam voltage is lower } } (resulting in poorer peak/bremmstrahlung ratios) and the emission of } x-rays } } from a solid sample is different from that in a thin foil, but these are } } some of the variables that Michael quite rightly pointed out must be } considered. } } } } 2: Comparison between EDX and WDX. } } } } This is like comparing apples and oranges, because the instruments } designed } } with them are generally intended for different purposes. } } } } WDX has a much better peak resolution than WDX, which results in better } } measured P/B ratios (and hence improved statistics), as well as much } better } } capability in resolving nearby x-ray lines. Also, because the x-ray } } counting and wavelength analysis are different functions in the crystal } } spectrometer, available countrates have traditionally been higher in WDX } } than EDX (although modern EDX detectors are an order of magnitude faster } } than they were fifteen years ago). Against this must be set the fact } that } } WDX is inherently a serial technique (although multiple spectrometers } help } } here), while EDX is a parallel technique (compare the advantages of PEELS } } over SEELS - although this is not a totally fair comparison). Anyway, } the } } advantage of WDX (ignoring the capability of resolving peak overlaps) is } } improved statistical precision. Is this useful? } } } } Well, maybe. } } } } By far the most significant parameter which affects electron-induced } x-ray } } emission spectra is sample geometry. Two extreme cases are where the } sample } } is a thin foil (as in the STEM) where, to a first approximation, the } x-ray } } spectrum recorded by the detector is the same as that emitted, and to a } } second order, it is possible to derive a reasonable thickness correction } for } } cases where the error is small. Alternatively, when the sample has } } dimensions large compared with the volume irradiated by the electron } beam, } } and has an accurately known geometry compared to the incident beam and } the } } detector (such as a polished flat sample in the microprobe), correction } } programs such as ZAF can do a reasonable job of extracting a quantitative } } analysis. } } } } What about where the sample geometry is unknown (for example, a rough } } surface such as might be examined in the SEM)? In that case the } uncertainty } } in the analysis is far, far worse than any uncertainty caused by the poor } } statistics of the EDX spectrum, so there is no point or advantage } whatever } } in using WDX to try to improve things, because it won't. This is why } } typically an SEM has an EDX detector - it is cheaper and gives just as } good } } an analysis (except for the somewhat poorer detection limit). In the } } microprobe, great care is taken in polishing and mounting the sample, so } the } } advantage of the WDX detector can be realised. } } } } Where does this leave us? The advantage of a WDX detector over an EDX } } detector *ON THE SAME SAMPLE* is limited - perhaps an order of magnitude } in } } detection limit, and, on a flat, polished sample, also perhaps } approaching } } an order of magnitude in precision. The WDX detector can also resolve } many } } cases where peaks would overlap in EDX. On general rough SEM samples, } the } } only advantages of WDX are a small improvement in detection limits and } the } } ability to resolve overlaps (which could be important if a trace element } } peak overlaps a major peak in the EXD spectrum. } } } } The President's science advisors were right - it all depends. I seem to } } remember that one of Jim Darley's Prime Ministers (Gough Whitlam?) had to } be } } dismissed because Parliament would not pass his budget - but does that } have } } anything to do with Science? } } } } Tony Garratt-Reed } } } } } } } Hi John and Michael et al: } } } I recall that one of your Presidents (Kennedy?) was looking for a one } } } handed science adviser; the ones he had always prevaricated "on the one } } } hand and on the other hand". } } } } } } Michael is quite right, detection limits vary greatly depending on } endless } } } factors. However, it is useful to have some figures as guidelines: } } } Considering say the 10 elements following sodium. Detection of these is } } } about best. } } } For these in EDX the limit for quantitative analysis is about 1%. } Detection } } } limit is about 0.1%. Increasing counts and counting times beyond the } } } customary 100 seconds at perhaps 2000cps will scarcely improve either } } } limit. } } } } } } WDX is near quantitative to its detection limit and that is at least two } } } orders of magnitude greater than is EDX. } } } } } } I'll enter correspondence only when it concerns errors in excess of five } } } orders of magnitude. } } } Cheers } } } Jim Darley } } } } } } ProSciTech Microscopy PLUS } } } PO Box 111, Thuringowa QLD 4817 Australia } } } Phone +61 77 740 370 Fax: +61 77 892 313 } } } Great microscopy catalogue, 400+ Links, MSDS } } } ************************ http://www.proscitech.com.au } } } } } } } John J. Bozzola wrote: } } } } } } } } } } Under ideal (and reasonably attainable conditions), what is the } } } } } detection } } } } } limit (in grams) for EDX and WDX? Thanks. } } } } } } } } The question you ask is not specific enough ... to many factors to } be } } } } considered with regard to which elements you are interested in, and } } } } (e.g.) ... how sensitive your specimen is to long count times and high } } } } beam currents ... ... ask again ... } } } } cheerios, shAf } } } } -- } } } } } } } } } Anthony J. Garratt-Reed } } MIT Room 13-1027 } } 77 Massachusetts Avenue } } Cambridge, MA 02139-4307 } } United States of America } } } } Ph: 617-253-4622 } } Fax: 617-258-6478 } }
I gave shipping regulations as an example, where regulations frequently do not make much sense. While shipping regulations are not directly a 'microscopy matter' they do affect us all. Clearly, I did not advocate flaunting any regulations; it's foolhardy for an individual and suicidal for a business. But we are free to discuss: Are those rules wise, cost efficient and appropriate. It appears Chuck Garber thinks they are, I beg to differ. I am only concerned with international airfreight and the IATA defined 'dangerous goods' regulations. To make shipping safer, regulations can extend to packaging requirements, maximum quantities shippable, cargo only aircraft requirements for some goods and 'antidote packing'. Generally, packing requirements are reasonable. Weight limits set to exempt 'dangerous goods' are few and generally they are too high. This means tiny quantities of not particularly dangerous goods require very expensive shipping. Antidote packing is basically not used. It makes sense to pack OsO4 in a tin with a little full cream powdered milk. Vapour from a cracked vial would be absorbed and it gives the lab a handy supply of that powder to keep for any laboratory OsO4 problems. We have shipped OsO4 in that manner for many years now. Packing in vermiculate is less effective than newspaper - which would at least absorb some of the vapour. The funniest thing is the paperwork required and this is largely used to justify the expense of DG shipments. Spot checks and appropriate fines for non-compliance would be a better preventative than lots of paper work. To wit: The ValuJet disaster did happen despite those rules. Dangerous goods, certainly on international routes, go mostly by passenger aircraft. Picture the site of a crash - who would scramble through all that useless paper and to what good? The worst is the expense: We work on airfreight from the US at US$10/kg for the gross weight of medium sized shipments. For DG that figure for generally larger shipments is US$25/kg - which frequently is more than the value of the goods. This cost (+paperwork and extra packing) would be the cost the end-user has to bear if it meant substantially improved safety. I can see none. The overall global cost of DG shipments above the cost of normal shipments has to amount to several billion $ annually. It suits IATA and the airlines and apparently Chuck Garber. I submit that we are getting negligible additional safety for thousands of megabucks. Retaining and improving on the rules for packing and marking of boxes but severely limiting the paperwork would be a rather more effective option. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 400+ Links, MSDS ************************ http://www.proscitech.com.au
---------- } From: Garber, Charles A. {cgarber-at-2spi.com} } Date: Saturday, 6 September 1997 8:07 } Jim Darley wrote: } ================================================= } But too many regulations are not wise - eg. the dangerous goods shipping } laws which appear designed to make things more expensive but not safer. } ================================================= } We should all be taking the HAZMAT reguations, no matter where we live, } seriously. I can not comment about the laws in Australia, but I have a very } high regard for the regulations promulgated by the U. S. Dept. of } Transportation (DOT) and IATA (for international shipments) as well as the } reasons behind them. And I can state, from first hand experience, that if } the regulators are presented with sound technical information for a } reguation to be changed, they will indeed listen (at least in the US) and } sometimes make changes. } } If anyone remains unconvinced, about the importance of adherence to all } HAZMAT regulations, keep in mind that the ValuJet disaster was caused by } someone not taking seriously these same regulations. } } While it is correct that one can incur almost unbelievalbly high shipping } costs for HAZMATs, this is not always the case. In many instances, such as } for the ordering of osmium tetroxide, if one orders "smart", they can do } their shipping at costs only nominally more than if the same weight of } tweezers, grids, or SEM mounts were being shipped. Now this would not apply } for everything (e.g. our SPI Dusters, for example) but it does apply for a } surprising number of HAZMATs routinely used in EM labs. We are also in } the process of reformulating some of our embedding kits so they too can be } shipped at the lower prices. } } We have tried to explain how a customer can "order smart" on our website, } click on "Hazardous Items(Good News and Bad News)". While savings in } shipping costs for domestic US customers are possible, the real } beneficiaries are foreign customers now no longer are restricted to the use } of air freight (which has associated with it high minimum charges). } } And while we are on this subject, just remember, don't ever try to take } HAZMATs in checked airline luggage to save some money, it is unconscienable } from a moral standpoint and puts at risk everyone on the plane flight. } } Chuck } } =================================================== } Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 } President 1-(800)-2424-SPI } SPI SUPPLIES FAX: 1-(610)-436-5755 } PO BOX 656 e-mail: cgarber-at-2spi.com } West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com } } } Look for us! } ############################ } WWW: http://www.2spi.com } ############################ } ==================================================
You ma want to define and echo this question to the Society of Ultrastructural Pathology's List Server at their web site http://sup.ultrakohl.com.
} } } maria lucia ribeiro caldas {caldasml-at-amcham.com.br} 09/07/97 07:16pm } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am having technical problems in EM skin biopsies. I would like to have a detailed protocol for processing skin biopsies.
I would never touch the Be window! If oil is present and you need to clean... CAREFULLY drip solvent acrosse the face to rinse the window. Be sure the solvent of choice not only will disolve the oil, but NOT the materials used to construct the Be window. Woody
{snip}
Let's go to the question: Have you ever cleaned the Be window of your EDS system? If the answer is positive, how often? Or, just wake me up ---- NEVER EVER THINK ABOUT THAT, YOU THE TROUBLE MAKER! I swear I have never been a trouble-maker. I just want to learn something from you. Just talking, no action following. It is too lang. Thank you for your time.
If you wish to be removed from this mailing list, please reply with the subject "Remove" and this software will automatically block you from future mailings.
Register Your Web Site To Over 590 Search Engines "Instantly". FREE DOWNLOAD.
I would like to inform you about an exciting, new, and award winning product that could both, save you countless hours, and increase your exposure on the Internet immeasurably! Known as the Submission Wizard, our program offers web users, such as yourself, the ability to automatically submit your site to over 590 search engines in just one session . Select from different categories, even countries, the choice is yours.
The submission process is very simple, efficient, and most importantly fast. All you do is provide standard site information i.e. site name, address, description, and a few well chosen keywords, the Submission Wizard takes this information changes it into the format required by each different engine, and registers your web site. It's that simple!
In addition, the Wizard's database of search engines and directories is constantly being updated every week, thus giving your site fresh and relevant exposure. Also, the Wizard allows you to register multiple URL's.
Just imagine the possibilities! Take charge of your web exposure, try out the Submission Wizard demo FREE by following the provided hyperlink. We know you'll be as excited about it as much as we are!
Want to be #1 on the search engines, check out our web page we will show how to do it with our new publication.
http://www.submissions.com/demo.html
NOTE: If you are using an Apple platform you will need windows emulation software.
Thank you for your time, D. Reynolds Customer Services.
I enjoyed your discussion on x-ray analysis and the challenge one faces from some students. As the saying goes, "you never really understand something until you teach it." Students will forever keep us on our toes and perhaps even force us to open certain doors that we might otherwise have tip-toed by.
Anyway, you asked does one clean the detector window? Yes, we clean our NORVAR window perhaps once a year or two - depending upon how dirty it has become. Most software has a subroutine for checking the efficience of the window using a standard of some sort (iron, for example). When efficiency drops, then one should carefully clean the window. In our case, we clean the window by removing the detector and slowly allowing 10-15 ml of methanol to flow over the window. The methanol is not directed at the window but onto the metal part of the housing about an inch away from the window. The methanol then gently flows over the window and washes away most of the oil. Check with your manufacturer, however, since different windows will require different treatments and/or solvents. You must be very gentle with the detector since the window and/or electronics will be damaged by mishandling. Definitely talk to a service person before doing this - if you are new to this procedure. We were, but now we feel comfortable cleaning the window.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing 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 ####################################################################
Morphometry (i.e. measurement of form) in the biomedical field is mostly done on thin sections either at the LM or the EM level. The methods of choice for obtaining morphometric estimates from=20 sections are those of stereology. The "classical" foundations of stereology including assumption-based methods are covered in:
Modern design-based methods, mainly developed during the last 10-15 years, try to focus on the theory of unbiased sampling and on the counting and sizing of particles as well as on the orientation of=20 structures in 3D space. Reviews of modern stereological methods are e.g.:
Gundersen HJG et al.: APMIS 96;379-394 and 857-881 (1988) Cruz-Orive LM, Weibel ER: Am J Physiol 258;L148-L156 (1990) Mayhew TM: Exp Physiol 76;639-665 (1991)
The Journal of Microscopy and Acta Stereologica are the official journals of the International Society for Stereology.
If you have any further questions please contact me directly.
Sincerely,
Matthias Ochs Matthias Ochs, M.D. Dept. of Anatomy Div. of Electron Microscopy University of G=F6ttingen Kreuzbergring 36 D-37075 G=F6ttingen Germany Phone: +49 551 397036 Fax: +49 551 397004
I'm forwarding this message for a colleague in my department. Please resond to me at the address below. TIA.
Owen
} Seeking appropriate polishing procedures for Ge-Sn samples } ---------------------------------------------------------- } } I have eight Ge-Sn samples ( five Ge-10 at% Sn and three Ge-40 at% Sn ) } which have been annealed at 400 C, 500 C, 600 C, 700 C, and 800 C for five } weeks. I need to have good polishing procedures so that good } polished surfaces can be obtained in order to provide a detailed microchemical } analysis on these samples. Sn smearing problems arise when a fine grid, for example, } 0.05 micron alumina, was applied to the samples. This might due to the } fact that we have both soft and hard materials in the samples. } } Any suggestion on how to polish these samples is highly appreciate. Thank } you.
Owen P. Mills Michigan Technological University Metallurgical & Materials Engineering Rm 512 MME Building Houghton, MI 49931 906-487-2002 906-487-2934 FAX opmills-at-mtu.edu
Does anyone over there know the address (fax, phone, whatever...) of the German maker of time programmators called DIEHL. We have been looking for them all around talking with various vendors of electronics material, not to avail. (if someone can tell me the www address of the phone numbers searching tool of Deutsche Telekom or Chamber of Commerce in Germany that could even be a hint).
Jennifer T. Morse wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } I was recently given the following homework assignment: Actual } resolution as determined by measuring the smallest distance between two } distinguishable points in a suitable specimen (resolution standard), is } usually greater than the linear resolution of the microscope. Describe } the factors that contribute to this discrepency. Find and list suitable } good references. Can you give me any help in solving this problem? Do } you know of any good sources to speak to? Do you know the answer? Thank } you for your help, Fred Meisenkothen
Jennifer & Fred,
Two of the best, upper level resources on this type of thing are: 1. Advanced Light Microscopy by Maksymilian Pluta (Elsevier - 3 volume set; volume 1 = ISBM 0-444-98939-0) 2. Progress in Microscopy, M. Francon (Row, Peterson & Co./Pergamon Press)
Born & Wolf's book on the Physics of Optics is also a good reference. I have no info on publisher, etc.
Let me know what you find.
Best regards, Barbara Foster Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com
RE73-at-aol.com wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } I am having trouble finding information concerning the theroy of morphometry } and its benifits to the biological field... any sugjestions.
Suggest the following: 1. John Russ's Handbook (already sent to you via another email) 2. Image analysis in Biology, Donat-P Hader, ed; CRC Press ISBN 0-8493-6033-1 3. Electronic Light Microscopy by David Shotton, ed., Wiley-Liss (ISBN 0-471-56077-4)
If you need just a quick overview of imaging principles, please see: Optimizing Light Microscopy for Biological and Clinical Laboratories, Kendall-Hunt (ISBN 0-7872-3538-5). We have them available, still on show discount from the Microscopy & Microanalysis meeting.
Good luck! Barbara Foster Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com
Your question is very broad so you will have to narrow the following list to your interests. Enjoy the search!:
Bibliography
Aherne, W.A. and M.S. Dunnill 1982 Morphometry. Edward Arnold (Publishers) Ltd. London,-205.
Bertram, J.F. and R.P. Bolender 1990 Counting cells with stereology: Random versus serial sectioning. J Electron Microsc Techn, 14:32-38.
Bianciardi, G., P. Tanganelli, and G. Weber 1993 Blood cell activation: New perspectives from ultrastructural morphometry. Semin Thromb Hemost, 19:108-114.
Bolender, R.P. 1978 Correlation of morphometry and stereology with biochemical analysis of cell fractions. Int Rev Cytol, 55:247-289.
Bolender, R.P. 1981 Stereology: applications to pharmacology. Annu Rev Pharmacol Toxicol, 21:549-573.
Bolender, R.P. 1982 Stereology and its uses in cell biology. Ann NY Acad Sci, 383:1-16.
Bolender, R.P. 1992a Quantitative morphology for biologists and computer scientists: I. Computer-aided tutorial for biological stereology (version 1.0). Microsc Res Techn, 21:338-346.
Cruz-Orive, L.-M. 1980 On the estimation of particle number. J Microsc, 120:15-27.
Cruz-Orive, L.-M. and E.R. Weibel 1981 Sampling designs for stereology. J Microsc, 122:235-257.
Cruz-Orive, L.-M. 1987 Particle number can be estimated using a disector of unknown thickness: the selector. J Microsc, 145:121-142.
Cruz-Orive, L.-M. and E.R. Weibel 1990 Recent stereological methods for cell biology: a brief survey. Am J Physiol, 258:L148-56.
Dardick, I. and D. Caldwell 1985 Reproducibility of morphometric image analysis. Hum Pathol, 16:1178
Freedman, L.S. 1974 A note on Aherne's method of counting tissue components in relatively thick sections. J Microsc, 100:219-225.
Gundersen, H.J.G. 1977 Notes on the estimation of the numerical density of arbitrary profiles: the edge effect. J Microsc, 111:219-223.
Gundersen, H.J.G., M. Boysen, and A. Reith 1981 Comparison of semiautomatic digitizer-tablet and simple point counting performance in morphometry. Virchows Arch B Cell Pathol, 37:317-325.
Gundersen, H.J.G. 1986 Stereology of arbitrary particles. A review of unbiased number and size estimators and the presentation of some new ones, in memory of William R. Thompson. J Microsc, 143:3-45.
Gundersen, H.J.G. and E.B. Jensen 1987 The efficiency of systematic sampling in stereology and its prediction. J Microsc, 147(3):229-263.
Gundersen, H.J.G. 1988 The nucleator. J Microsc, 151:3-21.
Hammel, I., D. Lagunoff, M. Bauza, and E. Chi 1983 Periodic, multimodal distribution of granule volumes in mast cells. Cell Tissue Res, 228:51-59.
Henderson, W.R. and E.Y. Chi 1985 Ultrastructural characterization and morphometric analysis of human eosinophil degranulation. J Cell Sci, 73:33-48.
Loud, A.V., W.C. Barany, and B.A. Pack 1965 Quantitative evaluation of cytoplasmic structures in electron micrographs. Lab Invest, 14:258-270.
Loud, A.V. and P. Anversa 1984 Biology of disease: Morphometric analysis of biologic processes. Lab Invest, 50:250-261.
Maser, M.D. 1990 An overview of morphometry and stereology. Tx A&M Univ EM Views, Issue No. 4:3-8,15-16.
Mathieu, O., L.-M. Cruz-Orive, H. Hoppeler, and E.R. Weibel 1981 Measuring error and sampling variation in stereology: comparison of the efficiency of various methods for planar image analysis. J Microsc, 121:75-88.
Mayhew, T.M. and M.A. Williams 1971 A comparison of two sampling procedures for stereological analysis of cell pellets. J Microsc, 94:195-204.
Mayhew, T.M. and L.-M. Cruz 1973 Stereological correction procedures for estimating true volume proportions from biased samples. J Microsc, 99:287-299.
Mayhew, T.M. and L.-M. Cruz-Orive 1974 Caveat on the use of the Delesse principle of areal analysis for estimating component volume densities. J Microsc, 102:195-207.
Mayhew, T.M. 1979 Basic stereological relationships for quantitative microscopical anatomy - a simple systematic approach. J Anat, 129:95-105.
Mayhew, T.M. and F.H. White 1980 Ultrastructural morphometry of isolated cells: methods, models and applications. Pathol Res Pract, 166:239-259.
Mayhew, T.M. 1981 On the relative efficiencies of alternative ratio estimators for morphometric analysis of cell membrane surface features. J Microsc, 122:7-14.
Nazeran, H., F. Rice, W. Moran, and J. Skinner 1995 Biomedical image processing in pathology: a review. Australas Phys Eng Sci Med, 18:26-38.
Peachey, L.D. 1982 A simple digital morphometry system for electron microscopy. Ultramicrosc, 8:253-262.
Pesce, C.M. 1985 Morphometric studies need review by statisticians. Am J Clin Pathol, 83:258
Pesce, C.M. 1987 Biology of disease. Defining and interpreting diseases through morphometry. Lab Invest, 56:568-575.
Schmid-Sch”nbein, G.W. and S. Chien 1989 Morphometry of human leukocyte granules. Biorheol, 26:331-343.
Sokal, R.R. and F.J. Rohlf 1981 Biometry: the principles and practice of statistics in biological research. Freeman, San Francisco,
Sokol, R.J., G. Hudson, J. Wales, and N.T. James 1988 Morphometry of human blood leukocyte ultrastructure: Its potential value in haematology. Haematol, 21:129-139.
Sokol, R.J., G. Hudson, J. Wales, and N.T. James 1991 Ultrastructural morphometry of human leucocytes in health and disease. Electron Microsc Rev, 4:179-195.
Sterio, D.C. 1986 The unbiased estimation of number and sizes of arbitary particles using the disector. J Microsc, 134:127-136.
Stoeber, W. 1965 Statistical size distribution analysis. Lab Invest, 14:892-908.
Underwood, E.E. 1970 Quantitative stereology. Addison-Wesley, Reading,
Vedel Jensen, E.B. and H.J.G. Gundersen 1992 The rotator. J Microsc, 170:35-44.
Webster, P. and G. Griffiths 1994 A novel method for mean cell volume estimation. J Microsc, 174:85-92.
Weibel, E.R., G.S. Kistler, and W.F. Scherle 1966 Practical stereological methods for morphometric cytology. J Cell Biol, 30:23-38.
Weibel, E.R. 1969 Stereological principles for morphometry in electron microscopic cytology. Int Rev Cytol, 26:235-302.
Weibel, E.R. 1972 The value of stereology in analysing structure and function of cells and organs. J Microsc, 95:3-13.
Weibel, E.R. and R.P. Bolender 1973 Stereological techniques for electron microscopic morphometry. In: Principles and techniques of electron microscopy: Biological applications. Vol. 3. M.A. Hayat, ed. Van Nostrand Reinhold Co. New York, pp. 237-296.
Weibel, E.R. 1974 Selection of the best method in stereology. J Microsc, 100:261-269.
Weibel, E.R. 1975 Quantitation in morphology: possibilities and limits. Beitr Pathol, 155:1-17.
Weibel, E.R. 1981 Stereological methods in cell biology: where are we--where are we going? J Histochem Cytochem, 29:1043-1052.
Weibel, E.R. 1982 Biomorphometry in physiological and pathological research. Acta Med Pol, 23:115-125.
Weibel, E.R. 1989 Measuring through the microscope: development and evolution of stereological methods. J Microsc, 155:393-403.
Weibel, E.R. 1991 Fractal geometry: a design principle for living organisms. Am J Physiol, 261:L361-9.
------------------------------------- Name: Charles Gilbert VOC:(704)355-5261 Carolinas Medical Center FAX:(704)355-8424 Dept of Pediatric Research PO Box 32861 Charlotte, NC 28232-2861
Alcan International Limited, Kingston Research and Development Centre, is seeking a Materials Characterization Technologist to work in the area of metallography and electron optics (SEM, TEM, Electron Microprobe) support.
You will be involved in a wide range of activities aimed primarily at: (a) characterizing the microstructures of Alcan's automotive and packaging alloys which, in turn, support the Company's product and process development work, and (b) determining the factors which influence the performance of Alcan's sheet products. You will also play a key role in the ongoing development of the laboratory's materials characterization techniques.
We are seeking an individual skilled in metallographic specimen preparation and the operation and maintenance of both optical and electron microscopes. As the perfect candidate, you have a college diploma or university degree in a materials science discipline, or the equivalent, coupled with experience in both optical and electron metallography and in the use of PC-based data acquisition and analysis software. In addition, you must have the ability to cope with numerous activities simultaneously and adapt to changing priorities.
We offer an excellent compensation package commensurate with experience and a first-class working environment.
Interested candidates may send a resume by 18 September 1997 to: Personnel Administrator, Alcan International Limited, Kingston Research and Development Centre, Box 8400, Kingston, Ontario K7L 5L9. Fax: (613) 541-2308
".....CdS nano-particles, at least the ones we have seen, those that would be thin enough to "see" through, are smaller than the smallest holes we have ever seen anyone able to make in a "lacy" film. So I think that there might have been some misinformation accidently posted about this awhile back (see Aug. 11 posting DUNNTEM-at-aol.com working for Ted Pella, Inc.) If I am wrong about this please correct me and set the record straight."
Our posting on this subject stated that nono-particles have a tendency to adhere to the inner surface of the holes in the lacey films - an ideal location for EM imaging. Obviously, since the holes in the lacey film are usually from 1 micron up in diameter, a 5nm particle will not lie across a hole.
Charles Garber went on to say:
"This simple reality is often times missed by people worried about film thickness (which in fact ought to not even be relavent in the case of a lacey or holey film, because after all, you are getting your information through the holes, not through the "lacy" areas). So the whole need to worry about "thickness" per se really ought to be, in the case of lacey films, a non-issue!"
We made no reference to the thickness of the lacey film. Obviously it is a "non-issue". In fact, we do not see that anyone brought that subject up except Charles Garber.
We feel that it is generous of the creator of this list to allow vendors to post information which includes the products they have that might solve a technical problem. It is, we think, misuse of the list to suggest that a vendor is posting misinformation or to promote one's own products by criticizing another's products.
Let's keep competitive sales techniques out of the microscopy list.
We have a CM12 TEM/STEM that is shut down at the moment because the +24 volt supply (located in the remote rack) does not want to start up. This causes the microscope to shut down immediately after you press the main power button. Remote bench testing of this supply with all the safety circuits in place does not help in our diagnosis. We checked the setup with the +5 volt supply, which is almost identical and it works.
If anyone has had similiar problems to the startup sequence of this supply which prevents the microscope from powering up I would be very interested in an email message from you. ie. before we spend $4500 (Can) for a new/rebuilt supply from Philips.
email directly if you desire: eoptics-at-mcmaster.ca
Thanks in advance
Fred
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
} Several experts have advised me that we should give up the } dependence upon the standardless analysis of EDS, although the others } showed strong evidence to prove that it is working well.
Yes, but why not standardize? You are lucky to have a couple of extremely good EDS systems, give them a chance to work properly.
} My former colleague, Bruce, suggested that oily detector window } might cause the problem for softer radiation such as Al-Ka. This message } struck a spark in my poor memory. I really saw somebody who dared to wipe } oil off the thin Be-window of a TN-5400 EDS using a cotton ball with a } drop of acetone. That was ten years ago. } Let's go to the question: Have you ever cleaned the Be window of } your EDS system?
I use EDS for quantitative analysis of minerals, I get results which are comparable to WDS (provocative statement, which I'm happy to defend), I have an old probe with a fairly dirty vacuum system, and I clean my window when the response to Na drops by about 50%, which takes about 6 months. Al drops, too, but not so much. I clean it by gently dribbling a stream of Freon over the whole end of the detector so that the stream doesn't play onto the window directly, but runs down over it. I use Freon because it is a wonderful solvent for oil but is pretty non-aggressive towards epoxies etc which may be used in the construction of the window. I wouldn't use acetone, too aggressive. When I run out of Freon I'll use light petroleum spirit ("ligroin", "petroleum ether").
And DO NOT physically touch the window with ANYTHING at all!!!!!!!!!
But why don't you just follow the recommendations of your detector manufacturer?
In fact, this leads me to a question which often comes to mind when I read some of the postings --- they contain questions which could so easily be answered by the manufacturer, such as the one a couple of days ago about sources of replacement detector windows.
Am I just lucky in the quality of support that I get from Oxford Australia (take a bow, Keith, Julie, and John, for an unsolicited testimonial and thank-you)?
cheers
Ritchie
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
Barbara Foster wrote: } } Jennifer T. Morse wrote: } } } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } -----------------------------------------------------------------------.} } } I was recently given the following homework assignment: Actual } } resolution as determined by measuring the smallest distance between two } } distinguishable points in a suitable specimen (resolution standard), is } } usually greater than the linear resolution of the microscope. Describe } } the factors that contribute to this discrepency. Find and list suitable } } good references. Can you give me any help in solving this problem? Do } } you know of any good sources to speak to? Do you know the answer? Thank } } you for your help, Fred Meisenkothen } } Jennifer & Fred, } } Two of the best, upper level resources on this type of thing are: } 1. Advanced Light Microscopy by Maksymilian Pluta (Elsevier - 3 volume } set; volume 1 = ISBM 0-444-98939-0) } 2. Progress in Microscopy, M. Francon (Row, Peterson & Co./Pergamon } Press) } } Born & Wolf's book on the Physics of Optics is also a good reference. I } have no info on publisher, etc. } } Let me know what you find. } } Best regards, } Barbara Foster }
I am looking to convert an old Nikon Diaphot TMD with a 80/20 side prism, to a 100/100% side port output. All that is involved is changing the side prism. We have the 80/20 kind, which we would gladly exchange at our expense with anyone in possesion of the 100/100 prism.
If interested please contact me to arrange the particulars.
Two requests. Firstly, would anyone know of a how to guide, including download sites for bringing spectra files from a TN-5500 into a PC? We need cabling, some sort of FTP on both sides and hopefully a spectra manipulation program. I can't help but think there are hundreds of microscopists who would find this useful.
Secondly, I am assembling a list of shareware useful to microscopists. I would like to assemble a list of programs, descriptions and download sites, which will be posted to the listserver (or sent out directly to anyone who requests it, if it gets too long). Should replies to this second request be posted, or come to me directly? I think if the shareware is generally useful to microscopists, a brief description, review and URL would be appropriate material for the listserver. How about it Nestor?
I am send to you information from "Morphology Digest" 1997, issue 5, from 8 september 1997 (Nl) about creation new sterelogy list in Calgary. I think that will be interesting for all.
---------- Forwarded message ----------
I have to prepare some yeast for SEM. The person who requested this work wants some pretty pictures of his yeast, Schizosaccharomyces pombe, for use in seminars. I have read up on some techniques to use but have two main queries: 1. To collect the yeast onto filter paper ( a method used in several publications), do I just drop a suspension of the yeast onto the filter paper? What sort of filter paper do I use? Is there a "better" way of collecting these cells such as settling them onto poly-l-lysine coverslips? 2. Is there a preferred fixative that works? The literature suggests a plethora of fixative cocktails! For TEM, I slam the cells onto a liquid nitrogen cooled copper mirror and process them via substitution in methanol and embed them in lowicryl HM20. We do not have an ESEM so please don't suggest I view them unfixed. Thanking you all in advance, this really is a great way of learning and sharing information! Sarah Ellis
We are interested in measuring the thermal conductivity of micron and submicron phases in a composite.Could any one tell me if SPM (Scanning Probe Microscopy) can be used to measure thermal conductivity and if this is a quantitative or purely qualitative technique ? Are the results relative or absolute ?
Your question is very broad so you will have to narrow the following list to your interests. Enjoy the search!:
Bibliography
Aherne, W.A. and M.S. Dunnill 1982 Morphometry. Edward Arnold (Publishers) Ltd. London,-205.
Bertram, J.F. and R.P. Bolender 1990 Counting cells with stereology: Random versus serial sectioning. J Electron Microsc Techn, 14:32-38.
Bianciardi, G., P. Tanganelli, and G. Weber 1993 Blood cell activation: New perspectives from ultrastructural morphometry. Semin Thromb Hemost, 19:108-114.
Bolender, R.P. 1978 Correlation of morphometry and stereology with biochemical analysis of cell fractions. Int Rev Cytol, 55:247-289.
Bolender, R.P. 1981 Stereology: applications to pharmacology. Annu Rev Pharmacol Toxicol, 21:549-573.
Bolender, R.P. 1982 Stereology and its uses in cell biology. Ann NY Acad Sci, 383:1-16.
Bolender, R.P. 1992a Quantitative morphology for biologists and computer scientists: I. Computer-aided tutorial for biological stereology (version 1.0). Microsc Res Techn, 21:338-346.
Cruz-Orive, L.-M. 1980 On the estimation of particle number. J Microsc, 120:15-27.
Cruz-Orive, L.-M. and E.R. Weibel 1981 Sampling designs for stereology. J Microsc, 122:235-257.
Cruz-Orive, L.-M. 1987 Particle number can be estimated using a disector of unknown thickness: the selector. J Microsc, 145:121-142.
Cruz-Orive, L.-M. and E.R. Weibel 1990 Recent stereological methods for cell biology: a brief survey. Am J Physiol, 258:L148-56.
Dardick, I. and D. Caldwell 1985 Reproducibility of morphometric image analysis. Hum Pathol, 16:1178
Freedman, L.S. 1974 A note on Aherne's method of counting tissue components in relatively thick sections. J Microsc, 100:219-225.
Gundersen, H.J.G. 1977 Notes on the estimation of the numerical density of arbitrary profiles: the edge effect. J Microsc, 111:219-223.
Gundersen, H.J.G., M. Boysen, and A. Reith 1981 Comparison of semiautomatic digitizer-tablet and simple point counting performance in morphometry. Virchows Arch B Cell Pathol, 37:317-325.
Gundersen, H.J.G. 1986 Stereology of arbitrary particles. A review of unbiased number and size estimators and the presentation of some new ones, in memory of William R. Thompson. J Microsc, 143:3-45.
Gundersen, H.J.G. and E.B. Jensen 1987 The efficiency of systematic sampling
in stereology and its prediction. J Microsc, 147(3):229-263.
Gundersen, H.J.G. 1988 The nucleator. J Microsc, 151:3-21.
Hammel, I., D. Lagunoff, M. Bauza, and E. Chi 1983 Periodic, multimodal distribution of granule volumes in mast cells. Cell Tissue Res, 228:51-59.
Henderson, W.R. and E.Y. Chi 1985 Ultrastructural characterization and morphometric analysis of human eosinophil degranulation. J Cell Sci, 73:33-48.
Loud, A.V., W.C. Barany, and B.A. Pack 1965 Quantitative evaluation of cytoplasmic structures in electron micrographs. Lab Invest, 14:258-270.
Loud, A.V. and P. Anversa 1984 Biology of disease: Morphometric analysis of biologic processes. Lab Invest, 50:250-261.
Maser, M.D. 1990 An overview of morphometry and stereology. Tx A&M Univ EM Views, Issue No. 4:3-8,15-16.
Mathieu, O., L.-M. Cruz-Orive, H. Hoppeler, and E.R. Weibel 1981 Measuring error and sampling variation in stereology: comparison of the efficiency of various methods for planar image analysis. J Microsc, 121:75-88.
Mayhew, T.M. and M.A. Williams 1971 A comparison of two sampling procedures for stereological analysis of cell pellets. J Microsc, 94:195-204.
Mayhew, T.M. and L.-M. Cruz 1973 Stereological correction procedures for estimating true volume proportions from biased samples. J Microsc, 99:287-299.
Mayhew, T.M. and L.-M. Cruz-Orive 1974 Caveat on the use of the Delesse principle of areal analysis for estimating component volume densities. J Microsc, 102:195-207.
Mayhew, T.M. 1979 Basic stereological relationships for quantitative microscopical anatomy - a simple systematic approach. J Anat, 129:95-105.
Mayhew, T.M. and F.H. White 1980 Ultrastructural morphometry of isolated cells: methods, models and applications. Pathol Res Pract, 166:239-259.
Mayhew, T.M. 1981 On the relative efficiencies of alternative ratio estimators for morphometric analysis of cell membrane surface features. J Microsc, 122:7-14.
Nazeran, H., F. Rice, W. Moran, and J. Skinner 1995 Biomedical image processing in pathology: a review. Australas Phys Eng Sci Med, 18:26-38.
Peachey, L.D. 1982 A simple digital morphometry system for electron microscopy. Ultramicrosc, 8:253-262.
Pesce, C.M. 1985 Morphometric studies need review by statisticians. Am J Clin Pathol, 83:258
Pesce, C.M. 1987 Biology of disease. Defining and interpreting diseases through morphometry. Lab Invest, 56:568-575.
Schmid-Sch”nbein, G.W. and S. Chien 1989 Morphometry of human leukocyte granules. Biorheol, 26:331-343.
Sokal, R.R. and F.J. Rohlf 1981 Biometry: the principles and practice of statistics in biological research. Freeman, San Francisco,
Sokol, R.J., G. Hudson, J. Wales, and N.T. James 1988 Morphometry of human blood leukocyte ultrastructure: Its potential value in haematology. Haematol, 21:129-139.
Sokol, R.J., G. Hudson, J. Wales, and N.T. James 1991 Ultrastructural morphometry of human leucocytes in health and disease. Electron Microsc Rev, 4:179-195.
Sterio, D.C. 1986 The unbiased estimation of number and sizes of arbitary particles using the disector. J Microsc, 134:127-136.
Stoeber, W. 1965 Statistical size distribution analysis. Lab Invest, 14:892-908.
Underwood, E.E. 1970 Quantitative stereology. Addison-Wesley, Reading,
Vedel Jensen, E.B. and H.J.G. Gundersen 1992 The rotator. J Microsc, 170:35-44.
Webster, P. and G. Griffiths 1994 A novel method for mean cell volume estimation. J Microsc, 174:85-92.
Weibel, E.R., G.S. Kistler, and W.F. Scherle 1966 Practical stereological methods for morphometric cytology. J Cell Biol, 30:23-38.
Weibel, E.R. 1969 Stereological principles for morphometry in electron microscopic cytology. Int Rev Cytol, 26:235-302.
Weibel, E.R. 1972 The value of stereology in analysing structure and function of cells and organs. J Microsc, 95:3-13.
Weibel, E.R. and R.P. Bolender 1973 Stereological techniques for electron microscopic morphometry. In: Principles and techniques of electron microscopy: Biological applications. Vol. 3. M.A. Hayat, ed. Van Nostrand Reinhold Co. New York, pp. 237-296.
Weibel, E.R. 1974 Selection of the best method in stereology. J Microsc, 100:261-269.
Weibel, E.R. 1975 Quantitation in morphology: possibilities and limits. Beitr Pathol, 155:1-17.
Weibel, E.R. 1981 Stereological methods in cell biology: where are we--where are we going? J Histochem Cytochem, 29:1043-1052.
Weibel, E.R. 1982 Biomorphometry in physiological and pathological research.
Acta Med Pol, 23:115-125.
Weibel, E.R. 1989 Measuring through the microscope: development and evolution of stereological methods. J Microsc, 155:393-403.
Weibel, E.R. 1991 Fractal geometry: a design principle for living organisms. Am J Physiol, 261:L361-9.
------------------------------------- Name: Charles Gilbert VOC:(704)355-5261 Carolinas Medical Center FAX:(704)355-8424 Dept of Pediatric Research PO Box 32861 Charlotte, NC 28232-2861
Cambridge Healthtech Institute's Advances in Cellular Imaging November 13-14, 1997 Westin Hotel Horton Plaza San Diego, California
TECHNICAL TRENDS AND ADVANCES Multiphoton Excitation Imaging and Photochemistry in Cells and Tissue Dr. Warren Zipfel, Cornell University Highly Resolved Cell and Tissue Optical Imaging in Real Time Dr. Daniel Farkas, Carnegie Mellon University Combined Fluorescent and Gold Cluster Probes: "Simultaneous" Labeling for Light and Electron Microscopy Dr. Richard Powell, NANOPROBES, Inc. Novel Magnetic Messenger Labeling System Mr. Lonnie Adelman, Ericomp Inc.
VIEWING REAL-TIME CELLULAR CHANGES Imaging Drug Uptake and Metabolism in Living Intestinal Tissue with Confocal and Two-Photon Microscopy Dr. Marshall Montrose, Johns Hopkins University School of Medicine Dynamic Changes in Intracellular pH and Ca2+ Dr Randi Silver, Cornell University Medical College (invited) Smart Magnetic Resonance Contrast Agents: A New Generation of Image Enhancement Media Dr. Tom Meade, California Institute of Technology Multiple Fluorescent Proteins and Fluorescence Microscopy to Monitor Live Cell Activity or Screen for Protein Localization Dr. Neal Gliksman, Universal Imaging Corporation Multiphoton Laser Scanning Fluorescence Microscopy Dr. Victoria Centonze Frohlich, University of Wisconsin- Madison
IMAGE ANALYSIS AND INTERPRETATION Quantitative Molecular Image Analysis Dr. Branko Palcic, British Columbia Cancer Research Centre (invited) Image Analysis Tools Dr. Paul Goodwin, Fred Hutchinson Cancer Research Institute (invited) Quantitative Automated Microscopy Dr. Frans Nauwelaers, Becton Dickinson Cellular Imaging Systems (invited) Fluorescence Imaging MicroSpectrophotometer (FIMS) Dr. Douglas Youvan, KAIROS Scientific Inc.
SCREENING AND DRUG DEVELOPMENT High-Content, Cell-Based Screening: Easing the Bottlenecks of Target Validation and Optimization of Lead Compounds Dr. Kenneth Giuliano, BioDx, Inc. Applications of the Fluorometric Imaging Plate Reader (FLIPR) Technology to High-Throughput Screening Dr. Simon Pitchford, Molecular Devices Corporation Use of Fluorescence Polarization in Drug Screening Dr. Michael Jolley, Jolley Consulting and Research Inc. (invited) Fluorescence-Based Screening of Cellular Changes in Ion Concentrations for Drug Development Dr. Carla Suto, SIBIA Neuroscience, Inc. (invited) Imaging Requirements for Faster Drug Development: Screening with Higher Density Formats Dr. Al Kolb, Packard Instrument Company (tentative)
Improved technology for imaging of cells and related targets is having a dramatic impact on pharmaceutical research and development, driven in part by the demand for greater speed, precision, and automation. Novel strategies for labels, better software for image enhancement and analysis, and progress integrating imaging with other laboratory functions are being applied to a growing range of applications. Advantages in such diverse segments as microscopy, cytology, and cellular analysis, as well as more applied uses such as assessment of gels and drug development screening, will be discussed. All of these activities share a similar goal of rapidly and correctly translating images into data that can be stored, used, compared, and manipulated with as much ease and as little human intervention as possible. These developments promise to have a dramatic impact on laboratory productivity, and any research manager involved in these segments should consider participating.
HOTEL INFORMATION Westin Hotel Horton Plaza Reservations made after the cut-off 910 Broadway Circle date will be accepted on a space and San Diego, CA 92101 rate availability basis. Available rooms are limited, so please book T: 619-239-2200 early. F: 619-239-0509 Please identify yourself as a Cut-off Date: October 30, 1997 Cambridge Healthtech Institute Room Rate: $135 Single/Double conference attendee to receive the reduced room rate.
TRAVEL INFORMATION TRAVELWORLD T: 717-288-9311 or 800-828-6033 601 Market Street F: 717-288-4693 Kingston, PA 18704
Exclusive airline discounts are available on American Airlines as well as other specific airlines when tickets are purchased through TRAVELWORLD at least 14 days prior to the meeting date. Some restrictions apply.
CALL FOR POSTERS Cambridge Healthtech Institute encourages attendees to gain further exposure by presenting their work in the poster sessions. Please fill out the registration form, with the poster title and primary author. To ensure inclusion in the conference binder, a one-page summary must be submitted by October 3, 1997.
CALL FOR EXHIBITORS Space is available for companies interested in exhibiting products and services related to cellular imaging. This meeting should attract up to several hundred senior researchers and managers representing a broad range of disciplines and perspectives. Please contact Jim MacNeil of Cambridge Healthtech Institute at 617-630-1341 to obtain an exhibitor package or to inquire about offering a workshop during the meeting. Exhibit space is limited so call now to reserve a space at this premier event.
Each registration includes all conference sessions, posters and exhibits, one luncheon and reception, continental breakfasts, all refreshment breaks, and a copy of the document binder.
Handicapped Equal Access: In accordance with the ADA, Cambridge Healthtech Institute is pleased to arrange for special accommodations for attendees with special needs. All requests for such assistance must be submitted in writing to CHI at least 30 days prior to the start of the meeting.
Substitution/Cancellation Policy In the event that you need to cancel a registration you may: Transfer your registration to a colleague within your organization. Credit your registration to another Cambridge Healthtech Institute program. Request a refund minus a $75 processing fee. Request a refund minus the cost ($150) of ordering a copy of the document binder. Cancellations will only be accepted up to one week prior to the conference.
Program and speakers are subject to change.
------------------------Cut and Print Here------------------------
Yes!|__| Please register me for Advances in Cellular Imaging 571E
Advance Registration (by October 3, 1997) |__| $795 Commercial |__| $395 Academic, Government, Hospital-Affiliated On-site or Late Registration (after October 3, 1997) |__| $895 Commercial |__| $445 Academic, Government, Hospital-Affiliated FIRST NAME:______________________________________________________ LAST NAME:_______________________________________________________ TITLE:___________________________________________________________ DIV./DEPT.:______________________________________________________ COMPANY:_________________________________________________________ ADDRESS:_________________________________________________________ City/State/ZIP:__________________________________________________ COUNTRY:_________________________________________________________ TELEPHONE:____________________________ Fax:______________________ E-MAIL:__________________________________________________________ |__| Please send information on exhibiting and opportunities to present workshops. |__| Enclosed is a check or money order payable to Cambridge Healthtech Institute, drawn on a U.S. bank, in U.S. currency. |__| Please charge: |__| AMEX (15 digits) |__| Visa (13 to 16 digits) |__| MasterCard (16 digits) Card #:___________________________________________________________ Exp. Date:________________________________________________________ Cardholder's Name:________________________________________________ Signature:________________________________________________________ Cardholder's Address (if different from above):___________________ __________________________________________________________________ |__| Reserve with credit card information listed above and invoice me. (Invoices must be paid in full by the deadline to retain registration discount. Invoices unpaid one week prior to conference will be billed to credit card at full registration rate.) If you plan to register on site, please check with CHI beforehand for space availability. |__| I am interested in presenting a poster at ADVANCES IN CELLULAR IMAGING and will provide an abstract by October 17, 1997. Poster title:______________________________________________________ ___________________________________________________________________
FAX or MAIL your reservation/registration to: Cambridge Healthtech Institute tel: 617-630-1300 1037 Chestnut Street fax: 617-630-1325 Newton Upper Falls, MA 02164 e-mail: chi-at-healthtech.com http:www.healthtech.com/conferences/
In a message dated 97-09-09 03:38:32 EDT, jbest-at-vicon.net (John Best) writes:
{ { Firstly, would anyone know of a how to guide, including download sites for bringing spectra files from a TN-5500 into a PC? We need cabling, some sort of FTP on both sides and hopefully a spectra manipulation program. I can't help but think there are hundreds of microscopists who would find this useful. } }
Greeting John,
We have just the program for you.
The software is called VIDX X-ray Microanalysis. It is a Windows 95 and NT software. (It is regularly sold to function with our PC based X-ray microanalysis and digital Imaging hardware)
The software will open fthe most popular file formats. We can perform both offline and on-line connections to many older vintage X-ray analyzers such as Edax, Kevex, Link, Noran, PGT, Tracor. That means you don't have to go and buy a brand new x-ray analyzer.
Prices are affordable.
For more information contact us at
Evex Analytical 857 State Road Princeton, NJ 08540 609-252-9192
Collect the yeast on to membrane filters with nice circular pores (e.g. Nucleopore), not a torturous-path filter like filter paper (or e.g. Millipore). This will give a nice smooth background against which to view the yeast. Filters with 0.22 micron holes are maybe best, although 0.45 micron will work. The pores will also give an *approximate!* size standard for the yeast cells.
*Before* collecting the yeast, coat both sides of the filters in a sputter coater. This gives better conductivity for viewing. If you're rich, use silver filters instead.
For fixation, I'd use the recipe the article(s) that show SEMs of yeast most like what you're trying to achieve (including 'scope kV), and is the simplest.
Phil
} I have to prepare some yeast for SEM. The person who requested this } work wants some pretty pictures of his yeast, Schizosaccharomyces pombe, } for use in seminars. I have read up on some techniques to use but have } two main queries: } 1. To collect the yeast onto filter paper ( a method used in } several publications), do I just drop a suspension of the yeast onto the } filter paper? What sort of filter paper do I use? Is there a "better" } way of collecting these cells such as settling them onto poly-l-lysine } coverslips? } 2. Is there a preferred fixative that works? The literature } suggests a plethora of fixative cocktails! For TEM, I slam the cells } onto a liquid nitrogen cooled copper mirror and process them via } substitution in methanol and embed them in lowicryl HM20. } We do not have an ESEM so please don't suggest I view them unfixed. } Thanking you all in advance, this really is a great way of learning and } sharing information! } Sarah Ellis
} 1. To collect the yeast onto filter paper ( a method used in } several publications), do I just drop a suspension of the yeast onto the } filter paper? What sort of filter paper do I use? Is there a "better" } way of collecting these cells such as settling them onto poly-l-lysine } coverslips?
You can drop the cells onto poly-l-lysine coated coverslips after osmium fixation. Then process it as usual for dehydration and critical point dry.
Best regards,
*********************************************** * Ming H. Chen, PhD * * Medicine/Dentistry Electron Microscopy Unit * * University Of Alberta. * * Edmonton, Alberta, Canada * * * * Visit My Page At: * * http://www.ualberta.ca/~mingchen * ***********************************************
Hello, Here is info on our procedures. We are a "Skin LAB".
PROCESSING OF SKIN TISSUE FOR LIGHT AND ELECTRON MICROSCOPY EMBEDDING IN EPON 812 1. Place newly received tissue* in 1/2 Karnovskys fixative overnight. 2. Take EPON 812 mixture out of refrigerator (let it sit under the hood for at least 2 hours before opening the container). This mixture is a combination of DDSA, NMA, and EMBED 812. (48% of 812, 31% of DDSA, 21% of NMA) 3. Rinse biopsy in 0.1 M sodium cacodylate buffer x2, 15 minutes each rinse. 4. Post-fix in 1% osmium tetroxide, 1 and 1/2 hours. (mix 1:1, 2% OsO4 with 0.2 M sodium cacodylate buffer). 5. Rinse in dH2O x 2, 15 minutes each rinse. 6. En-bloc stain with 1% Uranyl Acetate for 1 and 1/2 hours. 7. Dehydrate through an ascending ETOH series 35% x2 (15 minutes each) 70% x2 (15 minutes each) 95% x2 (15 minutes each) 100% x2 (30 minutes each) 8. Add the catalyst to the EPON 812 mixture (.2ml of DMP30 per 10 ml resin). Stir slowly for 10 minutes. 9. Clear biopsy in Propylene oxide x2, 15 minutes each rinse. 10. Infiltrate by placing biopsy into a 3:1 mixture of Propylene oxide:EPON for 3-4 hours 2:1 mixture for 12-16 hours (overnight) with caps off 1:1 mixture for 12-16 hours (overnight) with caps off 11. Place biopsy into an embedding mold with fresh 100% EPON for 8 hours, then place mold into 60 oC oven for curing (24-48 hours).
EPON 812 can be substituted with either EMBED 812 (EMS), PolyBed 812 (Polysciences), Medcast or Eponate 12 (Ted Pella). All have the ingredients DDSA, NMA, and DMP 30.
EMBEDDING PROCEDURE FOR SKIN BIOPSIES
ROUTINE RAPID
1/2 KARNOVSKYS overnight 2hr to overnight
0.1M NaCaco 15minx2 5-10min 1%OsO4/.1M Na Caco 11/2 hrs 20min
bake for 24-48 hours in 60oC oven DMP .2ml/10ml resin
Note: EPON 812 (which was discontinued in 1979) can be substituted with EMBED 812 (EMS), Polybed 812 (Polysciences), Eponate 12 or Medcast (TedPella). All have the ingredients DDSA, NMA, and DMP-30, plus one 'company specific ingredient'.
I pasted this in from Word, I hope it makes some sense.
Bob Underwood Morphology Core Univ. of Washington -
On Sun, 7 Sep 1997, maria lucia ribeiro caldas wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I am having technical problems in EM skin biopsies. I would like to have } a detailed protocol for processing skin biopsies. }
Your question is very broad so you will have to narrow the following list to your interests. Enjoy the search!:
Bibliography
Aherne, W.A. and M.S. Dunnill 1982 Morphometry. Edward Arnold (Publishers) Ltd. London,-205.
Bertram, J.F. and R.P. Bolender 1990 Counting cells with stereology: Random versus serial sectioning. J Electron Microsc Techn, 14:32-38.
Bianciardi, G., P. Tanganelli, and G. Weber 1993 Blood cell activation: New perspectives from ultrastructural morphometry. Semin Thromb Hemost, 19:108-114.
Bolender, R.P. 1978 Correlation of morphometry and stereology with biochemical analysis of cell fractions. Int Rev Cytol, 55:247-289.
Bolender, R.P. 1981 Stereology: applications to pharmacology. Annu Rev Pharmacol Toxicol, 21:549-573.
Bolender, R.P. 1982 Stereology and its uses in cell biology. Ann NY Acad Sci, 383:1-16.
Bolender, R.P. 1992a Quantitative morphology for biologists and computer scientists: I. Computer-aided tutorial for biological stereology (version 1.0). Microsc Res Techn, 21:338-346.
Cruz-Orive, L.-M. 1980 On the estimation of particle number. J Microsc, 120:15-27.
Cruz-Orive, L.-M. and E.R. Weibel 1981 Sampling designs for stereology. J Microsc, 122:235-257.
Cruz-Orive, L.-M. 1987 Particle number can be estimated using a disector of unknown thickness: the selector. J Microsc, 145:121-142.
Cruz-Orive, L.-M. and E.R. Weibel 1990 Recent stereological methods for cell
biology: a brief survey. Am J Physiol, 258:L148-56.
Dardick, I. and D. Caldwell 1985 Reproducibility of morphometric image analysis. Hum Pathol, 16:1178
Freedman, L.S. 1974 A note on Aherne's method of counting tissue components in relatively thick sections. J Microsc, 100:219-225.
Gundersen, H.J.G. 1977 Notes on the estimation of the numerical density of arbitrary profiles: the edge effect. J Microsc, 111:219-223.
Gundersen, H.J.G., M. Boysen, and A. Reith 1981 Comparison of semiautomatic digitizer-tablet and simple point counting performance in morphometry. Virchows Arch B Cell Pathol, 37:317-325.
Gundersen, H.J.G. 1986 Stereology of arbitrary particles. A review of unbiased number and size estimators and the presentation of some new ones, in memory of William R. Thompson. J Microsc, 143:3-45.
Gundersen, H.J.G. and E.B. Jensen 1987 The efficiency of systematic sampling
in stereology and its prediction. J Microsc, 147(3):229-263.
Gundersen, H.J.G. 1988 The nucleator. J Microsc, 151:3-21.
Hammel, I., D. Lagunoff, M. Bauza, and E. Chi 1983 Periodic, multimodal distribution of granule volumes in mast cells. Cell Tissue Res, 228:51-59.
Henderson, W.R. and E.Y. Chi 1985 Ultrastructural characterization and morphometric analysis of human eosinophil degranulation. J Cell Sci, 73:33-48.
Loud, A.V., W.C. Barany, and B.A. Pack 1965 Quantitative evaluation of cytoplasmic structures in electron micrographs. Lab Invest, 14:258-270.
Loud, A.V. and P. Anversa 1984 Biology of disease: Morphometric analysis of biologic processes. Lab Invest, 50:250-261.
Maser, M.D. 1990 An overview of morphometry and stereology. Tx A&M Univ EM Views, Issue No. 4:3-8,15-16.
Mathieu, O., L.-M. Cruz-Orive, H. Hoppeler, and E.R. Weibel 1981 Measuring error and sampling variation in stereology: comparison of the efficiency of various methods for planar image analysis. J Microsc, 121:75-88.
Mayhew, T.M. and M.A. Williams 1971 A comparison of two sampling procedures for stereological analysis of cell pellets. J Microsc, 94:195-204.
Mayhew, T.M. and L.-M. Cruz 1973 Stereological correction procedures for estimating true volume proportions from biased samples. J Microsc, 99:287-299.
Mayhew, T.M. and L.-M. Cruz-Orive 1974 Caveat on the use of the Delesse principle of areal analysis for estimating component volume densities. J Microsc, 102:195-207.
Mayhew, T.M. 1979 Basic stereological relationships for quantitative microscopical anatomy - a simple systematic approach. J Anat, 129:95-105.
Mayhew, T.M. and F.H. White 1980 Ultrastructural morphometry of isolated cells: methods, models and applications. Pathol Res Pract, 166:239-259.
Mayhew, T.M. 1981 On the relative efficiencies of alternative ratio estimators for morphometric analysis of cell membrane surface features. J Microsc, 122:7-14.
Nazeran, H., F. Rice, W. Moran, and J. Skinner 1995 Biomedical image processing in pathology: a review. Australas Phys Eng Sci Med, 18:26-38.
Peachey, L.D. 1982 A simple digital morphometry system for electron microscopy. Ultramicrosc, 8:253-262.
Pesce, C.M. 1985 Morphometric studies need review by statisticians. Am J Clin Pathol, 83:258
Pesce, C.M. 1987 Biology of disease. Defining and interpreting diseases through morphometry. Lab Invest, 56:568-575.
Schmid-Sch”nbein, G.W. and S. Chien 1989 Morphometry of human leukocyte granules. Biorheol, 26:331-343.
Sokal, R.R. and F.J. Rohlf 1981 Biometry: the principles and practice of statistics in biological research. Freeman, San Francisco,
Sokol, R.J., G. Hudson, J. Wales, and N.T. James 1988 Morphometry of human blood leukocyte ultrastructure: Its potential value in haematology. Haematol, 21:129-139.
Sokol, R.J., G. Hudson, J. Wales, and N.T. James 1991 Ultrastructural morphometry of human leucocytes in health and disease. Electron Microsc Rev, 4:179-195.
Sterio, D.C. 1986 The unbiased estimation of number and sizes of arbitary particles using the disector. J Microsc, 134:127-136.
Stoeber, W. 1965 Statistical size distribution analysis. Lab Invest, 14:892-908.
Underwood, E.E. 1970 Quantitative stereology. Addison-Wesley, Reading,
Vedel Jensen, E.B. and H.J.G. Gundersen 1992 The rotator. J Microsc, 170:35-44.
Webster, P. and G. Griffiths 1994 A novel method for mean cell volume estimation. J Microsc, 174:85-92.
Weibel, E.R., G.S. Kistler, and W.F. Scherle 1966 Practical stereological methods for morphometric cytology. J Cell Biol, 30:23-38.
Weibel, E.R. 1969 Stereological principles for morphometry in electron microscopic cytology. Int Rev Cytol, 26:235-302.
Weibel, E.R. 1972 The value of stereology in analysing structure and function of cells and organs. J Microsc, 95:3-13.
Weibel, E.R. and R.P. Bolender 1973 Stereological techniques for electron microscopic morphometry. In: Principles and techniques of electron microscopy: Biological applications. Vol. 3. M.A. Hayat, ed. Van Nostrand Reinhold Co. New York, pp. 237-296.
Weibel, E.R. 1974 Selection of the best method in stereology. J Microsc, 100:261-269.
Weibel, E.R. 1975 Quantitation in morphology: possibilities and limits. Beitr Pathol, 155:1-17.
Weibel, E.R. 1981 Stereological methods in cell biology: where are we--where are we going? J Histochem Cytochem, 29:1043-1052.
Weibel, E.R. 1982 Biomorphometry in physiological and pathological research. Acta Med Pol, 23:115-125.
Weibel, E.R. 1989 Measuring through the microscope: development and evolution of stereological methods. J Microsc, 155:393-403.
Weibel, E.R. 1991 Fractal geometry: a design principle for living organisms. Am J Physiol, 261:L361-9. ------------------------------------- Name: Charles Gilbert VOC:(704)355-5261 Carolinas Medical Center FAX:(704)355-8424 Dept of Pediatric Research PO Box 32861 Charlotte, NC 28232-2861
Someone recently commented about having a glassblower grind down a bell jar that has a chipped rim. We have had this done a number of times with good success. It is indeed a feasible solution, provided you can find a glassblower willing to undertake the task. On the other hand, you could probably do it yourself, if you are willing to devote the time and energy required, because all one glassblower I observed did was to spread a slurry of silicon carbide abrasive on a piece of window glass, and sit and rub the bell jar around over it.
Alternatively, we have been successful in some instances in 'patching' the chips in bell jars using a stiff epoxy (e.g. Torr Seal). Clean all grease and oil off the bell jar (try using Tilex Soap Scum Remover), then fill the chipped hole with the epoxy, and set the bell jar on a flat, smooth surface covered with waxed paper (so that the surface comes out flat and smooth) while the epoxy cures. Good luck,
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
Please define your problems more clearly. I have a lot of experience with skin biopsies, (2 published papers also) but I need to know what the problems are - embedding, cutting, staining for LM, wrinkles, etc. I would be happy to help if I had the details. Also, the size of the biopsy which you are required to handle will have a great influence on your final protocol. My handout at FORUM booth at the MSA meeting was an exact protocol which we use in our laboratory. I would be happy to send it to you, if I knew it would be of use in solving your particular problem. (Need your address).
} ... } Someone recently commented about having a glassblower grind down a bell jar } that has a chipped rim. We have had this done a number of times with good } success. ...
I consider a bell jar with any defect to be a considerable risk for an implosion.If your users don't use a implosion gaurd with 100% consistency then replace the bell jar ...
cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
} ... } Someone recently commented about having a glassblower grind down a bell jar } that has a chipped rim. We have had this done a number of times with good } success. ...
I consider a bell jar with any defect to be a considerable risk for an implosion.If your users don't use a implosion gaurd with 100% consistency then replace the bell jar ...
cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
Maria, We have worked with skin from a variety of species for over 20 years. We primarily work with human, pig, and in vitro equivalents. We do SEM, TEM, Immuno EM, enzyme histochemistry and basic light microscopy. We will be happy to share any of our techniques with you. What type of problem are you having? We use half strength Karnovsky's for fixation and embed in Spurr. Along time ago we used EPON 812 and then switched to Polybed and now Spurr. Sectioning with a diamond knife greatly enhances your sections.Please let us know your specific problems. Good Luck!!! NAMR
Nancy A. Monteiro-Riviere,Ph.D.,DABFE,DABFM Professor of Investigative Dermatology/Toxicology North Carolina State University College of Veterinary Medicine Cutaneous Pharmacology and Toxicology Center 4700 Hillsborough St. Raleigh, NC 27606 Telephone:(919)829-4426 FAX:(919)829-4358 email: Nancy_Monteiro-at-ncsu.edu CTPC Homepage:http://cptc.ncsu.edu
Allow me to second this repair method. I used it to repair a seriously chipped Denton 502A bell jar (it would work on any), and after overnight curing (room temperature), the unit pulled as good a vacuum as quickly as it did before the chip.
Phil
} Alternatively, we have been successful in some instances in 'patching' the } chips in bell jars using a stiff epoxy (e.g. Torr Seal). Clean all grease } and oil off the bell jar (try using Tilex Soap Scum Remover), then fill the } chipped hole with the epoxy, and set the bell jar on a flat, smooth surface } covered with waxed paper (so that the surface comes out flat and smooth) } while the epoxy cures. } Good luck, } } Wilbur C. Bigelow, Prof. Emeritus } Materials Sci. & Engr., University of Michigan } Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; } Fx:313-763-4788; Ph:313-764-3321
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Someone recently commented about having a glassblower grind down a bell jar } that has a chipped rim. We have had this done a number of times with good } success. It is indeed a feasible solution, provided you can find a } glassblower willing to undertake the task. On the other hand, you could } probably do it yourself, if you are willing to devote the time and energy } required, because all one glassblower I observed did was to spread a slurry } of silicon carbide abrasive on a piece of window glass, and sit and rub the } bell jar around over it. } } Alternatively, we have been successful in some instances in 'patching' the } chips in bell jars using a stiff epoxy (e.g. Torr Seal). Clean all grease } and oil off the bell jar (try using Tilex Soap Scum Remover), then fill the } chipped hole with the epoxy, and set the bell jar on a flat, smooth surface } covered with waxed paper (so that the surface comes out flat and smooth) } while the epoxy cures. } Good luck, } } Wilbur C. Bigelow, Prof. Emeritus } Materials Sci. & Engr., University of Michigan } Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; } Fx:313-763-4788; Ph:313-764-3321 } } We have repaired our Bell Jars 10-15 times using Vacuum epoxy, such as Bell Torr or Torr Seal. We do it a bit different than above. We actually file and sand the surface smooth. Basically, I have come to the conclusion that for the level of vacuum that Denton Coaters use, the only time a new bell needs to be bought is if it cracks. Chipping is easy to take care of.
oh...The reason I replied to the above is that if you use the wax paper technique, make sure you get the epoxy thick enough in the chip (ie. as thick as the glass)
In behalf of the "boss" of the EM core, I'll ask you for help... We like to know if there is somebody around who used LR white resins in an AFS from Leica. What are your conditoins, problems encountered etc... Thanks a lot for your help
Marc
PS Thank you Nestor for your work...
------------------------------ SCHMUTZ Marc PhD IGBMC 1 rue Laurent FRIES BP 163 F 67404 Illkirch Cedex FRANCE
I am a user of an old Coates&Welter/Nanometrics FEG-SEM and am interested in producing Electron Channeling Patterns. I need to rock the beam about a point on the sample for this but my microscope doesn't have this capability.
Are there any users (probably ex-users!) of this venerable machine who can tell me whether a beam deflection system was ever produced for the C&W or who have experience of ECPs with it? If so, where can i get hold of the necessary electronics?
Any help would be much appreciated.
ANGUS BEWICK Physics Dept. University of Bristol UK phab-at-siva.bris.ac.uk
This is a multi-part message in MIME format. --------------3F23D1509D80037EE77529EF Content-Type: text/plain; charset=us-ascii Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit Content-Transfer-Encoding: 7bit
Barbara Foster wrote:
} ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } ----------------------------- } -----------------------------------------. } Born & Wolf's book on the Physics of Optics is also a good reference. } I have no info on publisher, etc. } Let me know what you find. } } Best regards, } Barbara Foster } Microscopy/Microscopy Education } 53 Eton Street } Springfield, MA 01108 } PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com
Barbara:
I think you may mean:
Principles of Optics : Electromagnetic Theory of Propagation, Interference and Diffraction of Light by M. Born, E. Wolf Sixth Edition (Paperback) Published by Pergamon Press Publication date: June 1981 ISBN: 0080264816
-------------------------------------------------------------- Dr. Patrick L. Huddie (301) 725-2775 Fax (301) 725-2941 Microcosm, Inc., 9140 Guilford Road, Suite O, Columbia, MD 21046 e-mail phuddie-at-microcosm.com URL http://www.microcosm.com The Web is:"Vaster than empires and more slow" Andrew Marvell (1621-1678)
--------------3F23D1509D80037EE77529EF Content-Type: text/x-vcard; charset=us-ascii; name="vcard.vcf" Content-Transfer-Encoding: 7bit Content-Description: Card for Patrick Huddie Content-Disposition: attachment; filename="vcard.vcf"
begin: vcard fn: Patrick Huddie n: Huddie;Patrick org: Microcosm, Inc. email;internet: phuddie-at-microcosm.com title: CEO x-mozilla-cpt: ;0 x-mozilla-html: TRUE end: vcard
We are replacing our old SEM + EDS and would like dispose of it ourselves since the manufacturer of the new unit is offering only a nominal sum in a trade in. Can anyone suggest effective places to advertise this equipment? (We are located in NE USA, and would like to make room for the new system in 3-4 mo.)
A follow up question that I have is: what is a reasonable price to ask? ( The SEM is a 16 y.o. AMRAY 1600 turbo LaB6/W, secondary & Link backscatter, 2 CRT's, vibration isolation table, in good condition & under factory service contract. The EDS is 3 y.o. Oxford Isis thin window 136eV spec., 126eV MnKa calibrated, with beam control and image capture, under factory service contract. )
I will greatly appreciate any & all suggestions | comments.
Many thanks for your numerous and energetic inquiries about "Optimizing Light Microscopy for Biological and Clinical Labs"! Since our web site is not quite ready, in answer to those requests, we have put together a short order form (see below) as well as the description sent in an earlier email "re: infinity optics".
For those of you in the Northeast, you can also get a complimentary copy of the book by attending the one-day lecture-demo, "Optimizing Light Microscopy". Dates: Oct 3 (Providence College, Providence RI), Nov 3 (NYC), Nov 5 (Springfield, MA), Nov 7 (Boston, Tufts Med School MRC) Email for details.
Book Description: "Optimizing Light Microscopy for Biological and Clinical Laboratories" is available from MME as well as through the American Society for Clinical Laboratory Sciences and the publisher, Kendall-Hunt. Approximately 200 pages with over 100 diagrams, illustrations, and micrographs. Peppered with short experiments which are geared to help practicing microscopists learn more about how their light microscopes work. There are also introductory chapters on video microscopy and other microscopy techniques, ranging from Confocal to EM to microspectrometry.
To order, please download this form. If you are paying by check, just include the form with your payment. If you are paying by credit card (MasterCard and Visa accepted), just fax the completed, signed form back to the MME offices: (413)746-9311. Your book will be shipped within 7-10 business days.
Ordering Information:
Please note: These are special Microscopy & Microanalysis prices: Single copy: $30 + S/H Shipping/Handling: Please add $4.50 by Priority Mail within USA $3.35 by Book rate within USA $6.95 by Priority Mail to Europe Class-sized orders: $25 on orders of 25 or more shipped to one address, plus shipping (will be quoted at time of order)
Please send: ____ copies -at- $30 each = $_______________ Shipping/handling options: = $_______________ Book rate inside US: $3.25 Priority mail, inside US: $4.50 Priority mail, Europe: $6.95 (For shipments outside the US, please fax/email for details)
Total payment: = $ _______________
Method of payment: [ ] Check or money order (payable to MME) [ ] Charge my account: [ ] Visa [ ] MasterCard Acct #:_____________________________________________ Expiration date: ___________________________________ Billing address for card, if different from address above: Name: _____________________________________________ Address: __________________________________________ City/State/Zip: ___________________________________ Phone: ____________________________________________
SEND FORM TO: Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108 USA ` PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com
Many thanks for your order. We hope you enjoy "Optimizing Light Microscopy". Let us hear from you with successes and/or comments.
Barbara Foster Consortium President MME -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-Microscopy/Microscopy Education is a consortium of 24 consultants who specialize in customized on-site training in all areas of microscopy, sample preparation, and image analysis. Our goal: to help you use your microscope more effectively. -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
We are in the process of rebuilding our darkroom from the ground up, and were wondering if anyone had any information on suppliers of the revolving, light-tight darkroom doors and/or other darkroom fixtures.
Thanks.
Craig Lending Department of Biology SUNY Brockport Brockport, NY 14420
Torr Seasl is an epoxy compound especially formulated for use in vacuum systems that was sintroduced by Varian Associates, Vacuum Products Division, 121 Hartwell Ave, Lexington, MA 02173-3133 (800-882-7426) a number of years ago. It is stated to be good at pressures below 10-9Torr, and is bakeable at temperarures up to 120C. It adheres to most clean materials (glass, metals, ceramics) and holds up well over long term service. Some companies that handle EM supplies also handle it (e.g. I find it listed in the Ladd catalog).
Other similar products are also sold by other companies. For example, Duniway Stockroom Corp. (800-446-8811; info-at-duniway.com) sells a product called 'Epoxy Patch' that is stated to be equivalent to Torr seal
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
------------------------------------------------------- Dan Edwards Structural Materials Research Section Battelle Pacific Northwest National Laboratory P.O. Box 999, MSIN P8-15 Richland, WA 99352
Hello, We have a problem fixing imaginal discs (third instar - Drosophila). We are following a protocol used by Andrew Tomlinson, 1985- "The cellular dynamics of pattern formation in the eye of Drosophila" in J. Embryol. exp. Morph. 89, 313-331. The fixative used is a combined cold glutaraldehyde/osmium followed by an osmium post-fix. We're seeing pore fixation of membranes including inner cristae of mitochondria and some clear areas within cytoplasm. Thanks in advance for any suggestions. Rosemary
#################################################### Rosemary Walsh Electron Microscope Facility for the Life Sciences The Biotechnology Institute for Research and Education 1 South Frear Lab University Park, PA 16802 814-865-0212 email:rw9-at-psu.edu ####################################################
We supply Chromium Targets. If you would like to give us a call at 800/444-3137 and let us know what type of system you have, we will be happy to help you.
At least once over the history of this listserver there has been extensive discussion about osmium "pepper" and other artifactual inclusions seen in embedded, sectioned TEM samples. Does anyone have a compilation of replies/discussion or remember what conclusions were drawn? Answer directly please, to avoid boring others with a rediscussion, OK? Many thanks. Grace P.S. Was it related especially to phosphate buffered osmium solutions?? I just had a major disaster with some and would like to solve the problem quickly.....
The semiconductor industry uses "wafer" tweezers. They are used to pick up wafers from the side. The contact is not as minimal as the triceps, but they may work. There are some listed at the following URL. I am not sure where the ones we use around here came from. URL: http://www.ebsciences.com/labsupply/tweezers.htm
We have a student in our lab who is having a problem with her protoplasts that have been embedded in Epon/Araldite pulling away from the resin when she sections. This causes a lot of problems, to say the least. Do any of you have suggestions as to how to keep this from happening? She has heard that adding tannic acid to the fix helps prevent the problem. Has anyone done that? She's also having a problem with the inclusion bodies falling out of her sections when she cuts. Any suggestions about that? We recommended she reduce her cutting speed. All suggestions will be gratefully passed along to her.
Always eager to learn new things,
Paula = )
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
I have sent a repeated requests to resubscribe but without any results. I should be most grateful if you can put me on the list. If there are any problems, Please let me know.
Can anyone recomend a good, user friendly LM IM analysis software program? Our current one is useless (hate to say). We will be demoing a Noesis pacckage soon. I have experience with Optimas, and personally do not care for it. Any other shareware besides NIH? All comments etc welcome.
I posted the below earlier this week but the email bounced back. I believe that the the previously advanced concept of 'quantitative' analysis does require examination. JD.
I thank Garratt-Reed for corroberating my previous posting. John Bazzola had asked for a rough guide (he has confirmed that since) on detection limits of EDS versus WDS techniques. Anybody who has any significant experience with microprobe analysis knows that there is no single line correct answer. However, it is imperative for analysts to remember a few general figures so they can advise on appropriate instrumentation and techniques.
John B's initial inquiry deserved a reply and when none was given, I posted mine more than a day later. I believe that my posting is a useful guide for non-specialist analysts. Nothing that GR writes makes nonsence of my posting.
Nobody had asked about other differences between the techniques eg. resolution or simultaneous acquisition. I am pleased that G-R has supplied some information on those topics and on new, very high count-rate acquisition facilities for EDS. G-R is proud of his 0.6% "quantitative analysis" of Cr at an accuracy of +/- 0.1wt%.
I suggested that in EDS the presence of 1% of an element is the approxiamte lower limit for quantitative analysis. G-R has lowered that limit by some 40% - or has he? +/- 0.1% is good when 20% of the element is present, as it represents an accuracy of 0.5%. +/- 0.1% when 0.6% is present is about +/- 16% accuracy; I call that qualitative or at best semi-quantitative.
Another correspondent emailed me and noted that it was President Trueman who had been looking for a one handed adviser. But that was for an economist and not a scientist as I, apparently wrongly, remembered. The correspondent could see my point though; thanks to Brian Demczyk. I think that Trueman had an excellent idea but he should have extended that search to a scientist as well. Certainly microscopists and economists share disciplines which combine art and science. And I should add require 'good judgement'.
Why now was I abused with that opening: "Jim Darley's posting is hardly furthering good science". Am I to believe that good science is advanced by quarelsome nitpicking and that all broadbanding is verboten? I plead not guilty. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 400+ Links, MSDS ************************ http://www.proscitech.com.au
} } Subject: Re: detection limits x-ray } } Date: Friday, 5 September 1997 6:21 } } } } Jim Darley's posting is hardly furthering good science. } } } } 1: Detection limits (John's original question). Modern EDX systems are } } easily capable of quantitative analysis in the sub-1wt% range. See, for } } example http://prism.mit.edu/facltis/stem/stmexam.htm (the second } } illustration on that page) where I was getting analyses for Cr in steel } of } } the order of 0.6+/-0.1 wt%. I don't know about you, but I consider this } } quantitative. This, mind you, was in a measurement where I was } attempting } } to optimize spatial resolution rather than sensitivity, and the } acquisition } } time was 60sec. per data point. This example, of course, was from a } STEM. } } There are many more examples in the literature. } } } } Using beam gating techniques such as those developed by Charlie Lyman and } } colleagues, EDX data can readily be acquired at 20,000-40,000 counts per } } real second, if spatial resolution is sacrificed. Combine this with an } } acquisition time of 1,000 seconds (by no means unrealistic for an } important } } measurement) and the detection limit is in the rage of, or better than, } 0.01wt%. } } } } Of course, in the SEM, which may have been the point of John's original } } question, the situation is not the same, the beam voltage is lower } } (resulting in poorer peak/bremmstrahlung ratios) and the emission of } x-rays } } from a solid sample is different from that in a thin foil, but these are } } some of the variables that Michael quite rightly pointed out must be } considered. } } } } 2: Comparison between EDX and WDX. } } } } This is like comparing apples and oranges, because the instruments } designed } } with them are generally intended for different purposes. } } } } WDX has a much better peak resolution than WDX, which results in better } } measured P/B ratios (and hence improved statistics), as well as much } better } } capability in resolving nearby x-ray lines. Also, because the x-ray } } counting and wavelength analysis are different functions in the crystal } } spectrometer, available countrates have traditionally been higher in WDX } } than EDX (although modern EDX detectors are an order of magnitude faster } } than they were fifteen years ago). Against this must be set the fact } that } } WDX is inherently a serial technique (although multiple spectrometers } help } } here), while EDX is a parallel technique (compare the advantages of PEELS } } over SEELS - although this is not a totally fair comparison). Anyway, } the } } advantage of WDX (ignoring the capability of resolving peak overlaps) is } } improved statistical precision. Is this useful? } } } } Well, maybe. } } } } By far the most significant parameter which affects electron-induced } x-ray } } emission spectra is sample geometry. Two extreme cases are where the } sample } } is a thin foil (as in the STEM) where, to a first approximation, the } x-ray } } spectrum recorded by the detector is the same as that emitted, and to a } } second order, it is possible to derive a reasonable thickness correction } for } } cases where the error is small. Alternatively, when the sample has } } dimensions large compared with the volume irradiated by the electron } beam, } } and has an accurately known geometry compared to the incident beam and } the } } detector (such as a polished flat sample in the microprobe), correction } } programs such as ZAF can do a reasonable job of extracting a quantitative } } analysis. } } } } What about where the sample geometry is unknown (for example, a rough } } surface such as might be examined in the SEM)? In that case the } uncertainty } } in the analysis is far, far worse than any uncertainty caused by the poor } } statistics of the EDX spectrum, so there is no point or advantage } whatever } } in using WDX to try to improve things, because it won't. This is why } } typically an SEM has an EDX detector - it is cheaper and gives just as } good } } an analysis (except for the somewhat poorer detection limit). In the } } microprobe, great care is taken in polishing and mounting the sample, so } the } } advantage of the WDX detector can be realised. } } } } Where does this leave us? The advantage of a WDX detector over an EDX } } detector *ON THE SAME SAMPLE* is limited - perhaps an order of magnitude } in } } detection limit, and, on a flat, polished sample, also perhaps } approaching } } an order of magnitude in precision. The WDX detector can also resolve } many } } cases where peaks would overlap in EDX. On general rough SEM samples, } the } } only advantages of WDX are a small improvement in detection limits and } the } } ability to resolve overlaps (which could be important if a trace element } } peak overlaps a major peak in the EXD spectrum. } } } } The President's science advisors were right - it all depends. I seem to } } remember that one of Jim Darley's Prime Ministers (Gough Whitlam?) had to } be } } dismissed because Parliament would not pass his budget - but does that } have } } anything to do with Science? } } } } Tony Garratt-Reed } } } } } } } Hi John and Michael et al: } } } I recall that one of your Presidents (Kennedy?) was looking for a one } } } handed science adviser; the ones he had always prevaricated "on the one } } } hand and on the other hand". } } } } } } Michael is quite right, detection limits vary greatly depending on } endless } } } factors. However, it is useful to have some figures as guidelines: } } } Considering say the 10 elements following sodium. Detection of these is } } } about best. } } } For these in EDX the limit for quantitative analysis is about 1%. } Detection } } } limit is about 0.1%. Increasing counts and counting times beyond the } } } customary 100 seconds at perhaps 2000cps will scarcely improve either } } } limit. } } } } } } WDX is near quantitative to its detection limit and that is at least two } } } orders of magnitude greater than is EDX. } } } } } } I'll enter correspondence only when it concerns errors in excess of five } } } orders of magnitude. } } } Cheers } } } Jim Darley } } } } } } ProSciTech Microscopy PLUS } } } PO Box 111, Thuringowa QLD 4817 Australia } } } Phone +61 77 740 370 Fax: +61 77 892 313 } } } Great microscopy catalogue, 400+ Links, MSDS } } } ************************ http://www.proscitech.com.au } } } } John J. Bozzola wrote: } } } } } } } } } } Under ideal (and reasonably attainable conditions), what is the } } } } } detection } } } } } limit (in grams) for EDX and WDX? Thanks. } } } } } } } } The question you ask is not specific enough ... to many factors to } be } } } } considered with regard to which elements you are interested in, and } } } } (e.g.) ... how sensitive your specimen is to long count times and high } } } } beam currents ... ... ask again ... } } } } cheerios, shAf } } Anthony J. Garratt-Reed } } MIT Room 13-1027 } } 77 Massachusetts Avenue } } Cambridge, MA 02139-4307 } } United States of America } } } } Ph: 617-253-4622 } } Fax: 617-258-6478
Would anyone else like to help this soul. Send mail to his address (mluiselli-at-davinci.cnart.mx) not mine. Thanks
} Return-Path: {mluiselli-at-davinci.cnart.mx} } Date: Wed, 10 Sep 1997 18:33:42 -0700 } From: "Lic. Mariana Luiselli" {mluiselli-at-davinci.cnart.mx} } Organization: C N C A } To: sdw-at-biotech.ufl.edu } Subject: can you help me to find out what is K=F6eler lighting... } X-URL: http://www.biotech.ufl.edu/~emcl/tips.html } } In my homework they ask me, what is k=F6eler lighting, but i don=B4t know= =20 } what it is. Can you help me to find it out? } Thank you so very much, } mariana luiselli } }
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 352-392-1295 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
Thanks to all who responded to my CM12 +24 volt power supply problem.
The answer to the problem is to change C104 and C106, 47 ufd. 40 volts capacitors to 47 ufd. 63 or 100 volts. These capacitors are in the auxiliary power supply for startup, within the unit.
After the repair, I powered up the microscope and it worked the first time.
Thanks again Fred
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
Hello, I=B4m interested in staining protoplast with DAPI, but I don=B4t know how d= o=20 it.Is necessary a fixation?. I would like know a protocol to make this=20 staining. Best regards, M.D.Gomez email: gomezm-at-plantas.ibmcp.upv.es =20
If at all possible, could compilations of replies/discussion regarding osmium pepper be posted on the list server. I know that would serve useful for myself. Thanks.
Dan Caruso Biological Technician Medjet-at-worldnet.att.net
---------- } From: Grace Kennedy {kennedy-at-nsi.edu} } To: microscopy-at-Sparc5.Microscopy.Com } Subject: osmium pepper } Date: Wednesday, September 10, 1997 7:52 PM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } At least once over the history of this listserver there has been extensive } discussion about osmium "pepper" and other artifactual inclusions seen in } embedded, sectioned TEM samples. Does anyone have a compilation of } replies/discussion or remember what conclusions were drawn? Answer } directly please, to avoid boring others with a rediscussion, OK? Many } thanks. Grace P.S. Was it related especially to phosphate buffered } osmium solutions?? I just had a major disaster with some and would like to } solve the problem quickly..... }
At 04:13 PM 9/10/97 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Consolidated International Corp. 4501 South Western Blvd Chicago, IL 60609 800/621-3680 312/376-5600 312/376-5835 FAX
I don't know if there are other suppliers. Be very careful with the installation. We've had difficulties with ours primarily due to an incompetent installation.
} Can anyone recomend a good, user friendly LM IM analysis software program? } Our current one is useless (hate to say).
Which one is it?
} We will be demoing a Noesis pacckage soon.
Superb but not particularly friendly. Steep learning curve.
} I have experience with Optimas, and personally do not care } for it. Any other shareware besides NIH? All comments etc welcome.
ImagePro is probably the most friendly and is fairly powerful. For shareware, I strongly suggest you look at ImageTools (http://ddsdx.uthscsa.edu/dig/itdesc.html). An alternative which I liked quite a bit less but which seems powerful is Osiris (http://www.expasy.ch/www/UIN/html1/projects/osiris/ReadmeOsiris.html).
I want to supply Karnovsky (modified to 2.5% G and 2.5% "paraformaldehyde") to a lab to collect 5mm biopsies over a period of time. In the past I have used Karnovsky for up to a week after producing it but I am not sure about storing it for 1-2 months. Should I store at 4 deg C or consider freezing some (it will be in caodylate with 2.5mM CaCl2)?
Malcolm Haswell Electron Microscopy University of Sunderland
"Another correspondent emailed me and noted that it was President Trueman who had been looking for a one handed adviser. But that was for an economist and not a scientist as I, apparently wrongly, remembered. The correspondent could see my point though; thanks to Brian Demczyk. I think that Trueman had an"
I think this must have been a Freudian slip, and at certain times he might have even been called President Bluntman, but his name was "Truman".
Don Marshall
Donald J. Marshall Relion Industries P.O. Box 12 Bedford, MA 01730 Ph: 617-275-4695 FAX: 617-271-0252
I've been using the BioQuant system for about 5 years now and love it. It's sold by R&M Biometrics Inc. Phone 615-350-7866.
It's a Windows based program that is very versatile program, color recognition as well as grey scale. It can do sterography, cell counts (I automated my cell proliferation studies with it), etc.
the usual discliamer :) I have no connection with this Co. except as a statisfied customer. -- Begin original message --
} From: Microls-at-aol.com } } Can anyone recomend a good, user friendly LM IM analysis software program? } Our current one is useless (hate to say). We will be demoing a Noesis } pacckage soon. I have experience with Optimas, and personally do not care } for it. Any other shareware besides NIH? All comments etc welcome. } } Sincerely: } Lou Solebello } JM Huber Corp. } } }
-- End original message --
regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
I have just returned from holidays and have found a number of responses related to the question of gelatinized glutaraldehyde I had posed just before I left. Thanks to everyone for their interest. Oh by the way, I have now been able to entice one of my chemist colleagues to look at some of these samples to see whether she can discover a reason for this phenomenon as a result of differences in berry chemistry. If anyone is interested in the results, please contact me offline, but it will be awhile before the chemical analyses are done.
Thanks again.
Paula.
Paula Allan-Wojtas Food Microstructure Specialist Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia, Canada B4N 1J5
Dear Craig: Try Arkay Corporation, 228 South First Street, Milwaukee, Wisconsin Phone #1-800-TO-ARKAY; 414-276-9196. My catalogue shows that they carry both. Don Gantz Boston Univ. Med School
I would be interested to hear from Philips XL30 & XL40 users regarding how long their tungsten filaments last.
Thanks for your input. Nancy R. Smith Director of Operations Microscope And Graphical Imaging Center California State University, Hayward http://www.csuhayward.edu/SCI/sem
Wil Bigelow wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Torr Seasl is an epoxy compound especially formulated for use in vacuum } systems that was sintroduced by Varian Associates, Vacuum Products } Division, 121 Hartwell Ave, Lexington, MA 02173-3133 (800-882-7426) a } number of years ago. It is stated to be good at pressures below 10-9Torr, } and is bakeable at temperarures up to 120C. It adheres to most clean } materials (glass, metals, ceramics) and holds up well over long term } service. Some companies that handle EM supplies also handle it (e.g. I } find it listed in the LADD catalog). } } Other similar products are also sold by other companies. For example, } Duniway Stockroom Corp. (800-446-8811; info-at-duniway.com) sells a product } called 'Epoxy Patch' that is stated to be equivalent to Torr seal } } Wilbur C. Bigelow, Prof. Emeritus } Materials Sci. & Engr., University of Michigan } Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; } Fx:313-763-4788; Ph:313-764-3321
As a manufacturer of vacuum evaperators we have been plagued by chipping of bell jars for many years. We have used (and sold) Torr Seal and several other epoxies over the years and they are satisfactory for small chips. However, for large chips we have found that epoxy-putty GAPOX10(tm) works extremely well. It is inexpensive and cures in one hour for sanding and smoothing. We can pull a vacuum of 10(-7) and have yet to encounter a problem.
Thanks to all who responded to my osmium pepper question. I now know not to osmicate in phosphate but haven't decided what I should use for a buffer-I don't think phosphate and cacodylate are compatible so a simple switch may be out. Has anyone out there used Dalton's dichromate/osmium? I cannot find any reference to anything by Dalton. I do have a formula and have used it but am curious to know if anyone else has played with it. Tx Grace
} Thanks to all who responded to my osmium pepper question. I now know not } to osmicate in phosphate but haven't decided what I should use for a } buffer-I don't think phosphate and cacodylate are compatible so a simple } switch may be out. Has anyone out there used Dalton's dichromate/osmium? I } cannot find any reference to anything by Dalton. I do have a formula and } have used it but am curious to know if anyone else has played with it. Tx } Grace
In a former life, I used Dalton's fixative for immersion fixation of vertebrate retina. I don't remember all the details, but the results were very good. I don't remember this fix being widely used, though.
Hi All, does anyone know of a portable IR microscope vendor? A collleague of mine would like to purchase one to be able to look at sub surface features on silicon crystals as-grown. Since the crystals are rather large, it is easier to take the microscope to the crystal then vice-versa if possible.
I appreciate any help.
cheers
Lucio
Dr.Lucio Mule'Stagno MEMC Electronic Materials Inc University of Missouri -St.Louis Silicon Materials Research Group Physics Dept., 501 Pearl Dr., 8001 Natural Bridge rd., St.Peters, St.Louis MO 63376 MO 63121 tel: 314 279 5338 314 516 5931/3 fax: 314 279 5363 314 516 6152 email: lmulestagno-at-memc.com luciom-at-newton.umsl.edu
} Can anyone recomend a good, user friendly LM IM analysis software } program? Any other shareware besides NIH? All comments etc welcome. } } Sincerely: } Lou Solebello } JM Huber Corp.
ImageTool is a free image processing and analysis program for Windows 95/NT from the University of Texas Health Science Center at San Antonio.
http://ddsdx.uthscsa.edu/dig/itdesc.html
This list needs a FAQ.
-------------------------------------------------------------- Dr. Patrick L. Huddie (301) 725-2775 Fax (301) 725-2941 Microcosm, Inc., 9140 Guilford Road, Suite O, Columbia, MD 21046 e-mail phuddie-at-microcosm.com URL http://www.microcosm.com The Web is:"Vaster than empires and more slow" Andrew Marvell (1621-1678)
jss wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. }
} The Sparc5. Microscopy Station Master } unsubscribe
} Thanks to all who responded to my osmium pepper question. I now know not } to osmicate in phosphate but haven't decided what I should use for a } buffer-I don't think phosphate and cacodylate are compatible so a simple } switch may be out. Has anyone out there used Dalton's dichromate/osmium? I } cannot find any reference to anything by Dalton. I do have a formula and } have used it but am curious to know if anyone else has played with it. Tx
Grace, In our experience, osmium pepper is not caused by phosphate problems but by using aldehydes that have partially polymerized (e.g., old glutaraldehyde or formaldehyde solutions). The polymers are small enough to get into the cell but once attached to proteins, can not be removed. The polymers then vigorously reduce osmium which then shows up as pepper. I have routinely used phosphate buffered osmium and have never had the pepper problem. In fact, however, for osmication you really don't need to buffer at all. Distilled water is fine.
I used Dalton's chrome osmium many years ago for tissue culture cells and it worked fine. Now we prefer to use osimum ferrocyanide which gives better contrast. You can prep the solution using cacodylate. We fix in either cacodylate OR phosphate buffered glut/form, rinse extensively (overnight) in cacodylate buffer and then into the osmium.
If you need more info, contact me again as I can fax you the info for Dalton's.
John
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing 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 ####################################################################
It's been cute sitting in the corner, but now this prof. needs the space. Does anyone want a Philips 75 TEM, 1960s vintage? It's small, not too heavy, and actually ran until it blew a filament sometime back. I'm not sure which decade that was... I have the manuals and tools as well.
You would, of course, have to pay packing and shipping.
For more info, drop me a line at tina-at-pbrc.hawaii.edu
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
When taking a biopsy, careful attention to the following points will ensure that the histopathologist is quite unable to give any useful information.
1. Paint the skin with a strong antiseptic solution, preferably one that is deeply coloured (after all, the tissue has to be stained sooner or later).
2. Rub on the antiseptic vigorously to ensure that the surface layers of the epidermis are so disturbed as to be histologically uninterpretable.
3. Inject a considerable volume of local anaesthetic solution into the middle of the area to be biopsied and preferably inject the fluid rapidly. This will produce a satisfactory degree of tissue distension and distortion.
4. Seize the area to be biopsied with artery forceps, making sure that the forceps are clamped firmly enough to crush the tissue. Should the biopsy be too large to be effectively be dealt with by one pair of forceps then do not hesitate to use several pairs.
5. The tissue excised should be from the centre of the lesion and should not include any of the adjacent normal tissue. It is making things too easy for the Pathologist if the specimen allows the relationship of the lesion to the adjacent tissue to be seen.
6. When making the excision, two alternative and equally effective techniques may be considered. Either the biopsy can be only a fraction of a millimetre thick (thus saving the trouble of cutting sections) or a large piece of tissue can be excised. In this case make a number of tentative cuts so that the biopsy is partly cut through in several places. This will help to make the specimen impossible to orientate for proper sectioning.
7. If you think that the lesion may be an invasion tumour, always remove it piecemeal with a curette. The Pathologist will then probably not be able to tell whether it is invasive or not.
8. Before placing the tissue into fixative, ensure that is adequately covered with blood. This should be done so effectively that after fixation the specimen is entirely concealed within clot. This ensures that the Pathologist is kept busy trying to find the specimen and provides a reasonable chance that every section will be covered with red blood cells dislodged from the clots during the sectioning process.
9. If the biopsy is to be really uninterpretable, care must now be exercised in the selection of the container and the fixative. The container should be very small so that there is room for a minimum of fixative. Plastic sequestrene bottles (pink label) are admirable for this purpose. It is quite surprising how much tissue can be compressed into one of these. Alternatively, use a container with a very narrow neck. A mass of tissue is often quite soft when freshly excised and can be forced through a small hole. When fixed the tissue can not be removed from the bottle without breaking the glass. With luck fragments of glass will then be driven into the tissue. This lends excitement to the process of section cutting.
10. The fluid into which the specimen is placed should on no account be a conventional histological fixative, such as 10% neutral-buffered formol saline. Plain water or physiological saline will ensure adequate breakdown of the cells. If available, Stuart's transport medium is even more effective in producing putrefaction. If such a culture medium is used, the specimen should be kept on a radiator until dispatch and should be sent to the laboratory by post (this is especially effective during the summer months).
11. On no account should the container be labelled with the patient's name or any other mark of identification. If you can arrange for several unlabelled containers to arrive in the laboratory on the same day so much the better. There is an excellent chance that the specimens from different patients will be confused. An alternative, and equally effective, measure is to place more than one biopsy in the same container. If they are from the same patient make sure the pieces of tissue are all of a similar shape and size. Thus, if one biopsy shows a neoplasm no-one will know which of the biopsy sites contains the tumour.
12. In the accompanying request form on no account should you provide the Pathologist with the name of either the patient, yourself or your address. Otherwise, there will be some means of indexing the specimen and of knowing where to send the report when eventually prepared.
13. So that the Pathologist shall not be influence or biased in interpretation, avoid giving any information regarding the age of the patient, or the site of the biopsy, or the duration and appearance of the lesion. Above all, never mention your own differential diagnosis.
Finally, one or more of the following stratagems should be used in selected cases:
(a) Ask for serial specimens, bearing in mind that a specimen 5 mm thick will yield about a 1000 sections.
(b) Telephone the laboratory frequently for the report. If possible arrange for the first two calls to reach the laboratory before the specimens.
(C) Ask for rapid-frozen sections especially on large heavily-calcified masses that have been present for several years.
Dr. Elaine Humphrey Biosciences Electron Microscopy Facility University of British Columbia 6270 University Blvd Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-unixg.ubc.ca
HASWELL Malcolm wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I want to supply Karnovsky (modified to 2.5% G and 2.5% "paraformaldehyde") } to a lab to collect 5mm biopsies over a period of time. } In the past I have used Karnovsky for up to a week after producing it but I } am not sure about storing it for 1-2 months. Should I store at 4 deg C or } consider freezing some (it will be in caodylate with 2.5mM CaCl2)? } } Malcolm Haswell } Electron Microscopy } University of Sunderland
Malcolm:
I have stored this fixative in glass at room temperature for extended periods (1-2 months) and have had no problems. Once you see some bottom ppt. or cloudiness, discard with copious amounts of water. Also be careful of the formalin conc. in the air. Best regards, Jerome.
Dalton's dichromate-osmium fixative brings back some nostalgic memories. I used it to fix rat testicular Leydig cells for my PhD thesis research (about 1956-58, Harvard Biology), and the results suggested that the smooth endoplasmic reticulum was tubular, rather than than vesicular, as was commonly believed at the time. In my subsequent postdoctoral work with Don Fawcett, opossum Leydig cells fixed with Dalton's dichromate osmium exhibited tubular smooth ER, while the same material fixed with osmium-acetate veronal, commonly used at the time, had vesicular SER. This work was published in 1961 (Christensen, Fawcett 1961, J Biophys Biochem Cytol 9:653-670).
The reference for the fixative is an abstract: Dalton, A.J. 1955, "A chrome-osmium fixative for electron microscopy," Anat Rec 121:281. I knew Jack Dalton, who was at the National Cancer Institute, at NIH in Bethesda. He and Marie Felix were the first to show the Golgi complex by EM (1954, as I remember).
Kent
A. Kent Christensen Department of Anatomy and Cell Biology Medical Sciences II Building University of Michigan Medical School Ann Arbor, MI 48109-0616 akc-at-umich.edu Tel (313) 763-1287 http://www-personal.umich.edu/~akc/
----------------------------
On Thu, 11 Sep 1997, Grace Kennedy wrote:
} Thanks to all who responded to my osmium pepper question. I now know not } to osmicate in phosphate but haven't decided what I should use for a } buffer-I don't think phosphate and cacodylate are compatible so a simple } switch may be out. Has anyone out there used Dalton's dichromate/osmium? I } cannot find any reference to anything by Dalton. I do have a formula and } have used it but am curious to know if anyone else has played with it. Tx } Grace
Our lab is using the fluorescent antibody technique to detect and count Cryptosporidium and Giardia in water samples. One of the biggest interference?s with this is the auto-fluorescence of algal cells which is often brighter than the FITC stained cells we are looking for. This is particularly inconvenient as we are attempting to automate the microscopy with the use of Image analysis. Part of our efforts are going into producing a cleaner final concentrate but it is not always possible to eliminate all the algae. In view of this has any one come across a product/method for quenching auto-fluorescence that does not affect the FITC stain.
Phil Dobson Australian Water Quality Centre Hodgson Rd, Bolivar South Australia 5118 Phone 61 8 8259 0341 Fax 61 8 8259 0228 Email phil.dobson-at-sawater.sa.gov.au
I'm translating a microscope manual from Japanese into English and constantly running into sentences that go like "part of the field of view may be cut off" and "image cut-off in peripheral part may occur."
Could anyone tell me how to put them correctly? Should I instead say "the field of view may be vignetted" and "image vignetting may occur" which sound more technical to me but I'm not sure about.
Any suggestion is welcomed. Thanks in advance.
Chiba Atsushi [(Mr.) -- *Chiba* is my surname] Voice: (+81) 010-045-9451
have you considered measuring the emission at two wavelengths? The autofluorescence of algae should be pretty red, whereas fluorescein is predominantly green. Subtracting a scaled red image from the corresponding green image should leave you with only fluorescein fluorescence, presuming that all of the algae have the same fluorescence spectrum. Alternatively, if the cells remain discrete in the image, it would be possible to get your image analysis package to ignore the algae on the basis of size and/or create a mask in the red image that prevents the algae being counted in the green one.
Ray
At 3:39 pm +0930 12/9/97, Phil Dobson wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Ray Hicks ________________________________________________________________________ |University of Cambridge |Tel 01223 330149 | |Department of Medicine |Fax 01223 336846 | |Level 5, Addenbrookes Hospital |e-mail {rh208-at-cus.cam.ac.uk} | |Hills Road Cambridge |Web http://facsmac.med.cam.ac.uk | |CB2 |ftp server ftp://131.111.80.78 | |UK | | |_________________________________|_____________________________________|
We currently have the first version of the Buehler image analysis system and I have nor problems with it. It is a very powerful package and for all of our Materials analysis needs it performs very well. We have not upgraded to the latest version wich is supposed to be even better due to lack of funds. They will be happy to demo it for you if you give them a call.
Roberto Garcia EMF Manager Wright State University
Sorry about keeping you waiting. Bernies Photo Center can be reached at 800 346-8884. They will have verything you need and if I could make a suggestion splurge on a sodium vapor light, it is worth it. No more stumbling around the darkroom. They also have the best prices around on bulk 4X5 film. Good luck with the darkroom.
Roberto Garcia EMF Manager Wright State University Dayton, OH
In a message dated 97-09-12 08:45:19 EDT, Chiba writes: { { sentences that go like "part of the field of view may be cut off" and "image cut-off in peripheral part may occur." Could anyone tell me how to put them correctly? Should I instead say "the field of view may be vignetted" and "image vignetting may occur" which sound more technical to me but I'm not sure about. } }
The phrase "part of the field of view may be cut off" sounds ok to me. Without an illustration, I do not know what it means. My mental image is that there is a black part within a photographic image taken with the microscope from a flash/shutter timing problem, or from blocking of the field of view by an accessory that is inserted in the optical path, such as the quarter wavelength plate typically used in a polarizing microscope.
The phrase "image cut-off in peripheral part may occur." could be reworded as "...the peripheral part of the image may be cut-off." As above, I do no know really what it means. But I imagine that it means the effect from closing down the sub-stage iris. I do not think that is really vingetting, a gradual shading, but it is close to it. If the effect being discussed is from gross misalignment of the bulb in the illuminator housing, then it could be vignetting.
Steve Stokowski Stone Products Consultants Concrete Petrographers http://members.aol.com/CrushStone/index.htm
} Sorry for my layman's question. } I'm translating a microscope manual from Japanese into } English and constantly running into sentences that goes } like "part of the field of view may be cut off" and } "image cut-off in peripheral part may occur." } } Could anyone tell me how to put them correctly? } Should I instead say "the field of view may be } vignetted" and "image vignetting may occur" which sounds } more technical to me but I'm not sure about. }
I would not use the term "vignette" myself. It is a technical term that is used in lens design to mean that you have blocked part of a bundle of rays (hopefully on-purpose to kill the wild rays that would have not been well focussed, but sometimes by mistake because a lens element was too small). Although the word could be used for your purpose, few people know what it means, and some of them may be confused because of the technical use. I would just say "the field of view may be reduced," and "part of the field of view may be cut off." Making it sound more technical is helpful only for people with trouble falling asleep :)
best regards mark
Mark W. Lund, PhD Director } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"The state is good at simple tasks, like killing people and seizing their wealth. It has far more trouble reaching inside individuals and making them good." Doug Bandow
} Sorry for my layman's question. } I'm translating a microscope manual from Japanese into } English and constantly running into sentences that goes } like "part of the field of view may be cut off" and } "image cut-off in peripheral part may occur." } } Could anyone tell me how to put them correctly? } Should I instead say "the field of view may be } vignetted" and "image vignetting may occur" which sounds } more technical to me but I'm not sure about. }
I would not use the term "vignette" myself. It is a technical term that is used in lens design to mean that you have blocked part of a bundle of rays (hopefully on-purpose to kill the wild rays that would have not been well focussed, but sometimes by mistake because a lens element was too small). Although the word could be used for your purpose, few people know what it means, and some of them may be confused because of the technical use. I would just say "the field of view may be reduced," and "part of the field of view may be cut off." Making it sound more technical is helpful only for people with trouble falling asleep :)
best regards mark
Mark W. Lund, PhD Director } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"The state is good at simple tasks, like killing people and seizing their wealth. It has far more trouble reaching inside individuals and making them good." Doug Bandow
I have been trying several stains on fixed monolayer fibroblasts. The cells have been fixed by either formalin, formalin fumes, or gluteraldehyde. None of the stains are penetrating the cells. Does anyone have a suggestion?
We just bought a new Olympus inverted microscope for epi-fluorescence. We were very surprised to find that our 1.2NA water immersion objective gave BRIGHTER pictures and SHARPER pictures than our 1.4 NA oil immersion objective. We thought it was a problem with the 1.4 NA objective so the sales representative brought by:
1) another 1.4 NA oil immersion objective: whose pictures were as poor as the previous objective 2) a 1.25 NA oil immersion UV objective: whose bright spots were brighter, whose black regions were darker, and whose contrast was sharper than on the 1.4 NA objective. (The 1.25 NA objective is also much more affordable objective).
Does anyone have an explanation? One possibility was that the 1.4 NA objective is NOT a UV objective: we thought that some UV light may be coming through our 460-490 nm excitation filter causing auto-fluorescence in the objective. So we put another very sharp 488 nm filter in front of it and it did not resolve the problem.
In comparing the 1.4 NA oil to the 1.2 NA water objective, shouldn't the signal be 85% brighter? - on the excitation side: (1.4/1.2)2=1.36, or 36% more transmission of excitation - on the emission side: (1.4/1.2)2=1.36, or 36% more collection of emission in total: (1.4/1.2)**4=1.85.
So how could the lower NA objectives give brighter sharper signals?
We are collecting our images with a Hamamatsu Cooled-CCD camera. Both the shutter on the excitation and the collection are under software control, so we know that data from both objectives are collected under the same condition. We've used a series of test slides so each objected could be used on the same image. Any suggestions are welcome.
Thanks,
Sanford M. Simon Laboratory of Cellular Biophysics Box 304 Rockefeller University 1230 York Avenue New York, N.Y. 10021 212-327-8130 (voice) 212-327-8022 (fax) simon-at-rockvax.rockefeller.edu (e-mail)
In regards to (i) EM immunocytochemical staining on primary antibodies and collodial gold and (ii) uranyl acetate and Pb citrate, I would like to poll the list to determine how many people either float their EM grids or immerse them in droplets so that both sides are stained. I have always floated on a 20 ul drop but have started wondering if I am missing half the action. Comments about advantages and disadvantages welcomed!
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
I have a couple of questions and will appreciate any replies and suggestions.
1) What is the best sample to test the resolution of a TEM at a tilt angle (} 30degree)?
I am testing the resolution of a CM300FEG microscope. I used oriented gold, evaperated gold particles, graphitized carbon and so on. There is no problem of those samples at zero tilt. But after tilt the specimen to 30 degree or higher, I got only the good resolution (2.3A in case of evaperated gold particle) in one direction, along the tilt axis. I believed it is not because of the focus gradient that I could not get the same resolution in both directions at this tilt angle. I think the samples (evaporated gold particles and graphitized carbon) may not be the suitable samples for the resolution test of the high tilt angles (higher than 45 degree and up to 60 degree). I wonder if any of you has done such test before and has any suggestions on which specimen should be used in this test and where to get it?
2) Is there any published documentation about the line resolution of Kodak SO-163 film?
I am writing an paper about the performance of the CM300FEG at low magnificaiton, and need to know the line resolution of Kodak SO-163 (something I can cite for). But I could not find any published documentation. Of course, I called Kodak technique support and of course they didn't give me a clue. There is a web site called "Bibliography on EM Imaging and Related Technologies" (http://www.uct.ac.za/depts/emu/imaging/papers.htm), but I can not find any thing there either about SO-163. The only other related information I got was from a similar discussion in this board a year ago. But the data about line resolution found in that discussion were mostly estimated but not from any published documentation. I wonder if any of you know there is any kind of published documentation including research paper, technique report or so which gives the line resolution of Kodak SO-163. (BTW, the topic about the line resolution has been discussed a year ago and I am sorry to ask it again.)
I appreciate any information or suggestion on my request.
Yifan Cheng
BTW, I am not sure if my email address is already on the list or not. So that I appreciate also that when you reply this email, send a reply to me directly as well as to the board. Thanks again.
-- ********************************************************************** * Dr. Yifan Cheng * Phone: +1-850-644-4104 (Office)* * Institute of Molecular Biophysics * or: +1-850-644-9769 (Lab) * * Florida State University * Fax: +1-850-561-1406 * * Tallahassee, FL 32306-4380 * Email: ycheng-at-sb.fsu.edu * * U.S.A. * http://www.sb.fsu.edu/~ycheng * **********************************************************************
We have a Zeiss DSM-960 with LaB6. This system has a long history of problems with the column isolation valve (manually operated) leaking during specimen exchanges. Numerous fixes have been tried including slightly larger and softer o-rings and metal shims to change the height/pressure which the gate cams to when closed, all to no avail. Has any one experienced similar problems on a Zeiss or on other instruments and was a fix found.
Thanks, Jim Mabon _____________________________________________________ James C. Mabon Center for Microanalysis of Materials Frederick Seitz Materials Research Laboratory 104 South Goodwin Avenue Urbana, Illinois 61801 (217)333-4265 *Fax(217)244-2278 email: mabon-at-uimrl7.mrl.uiuc.edu {that's the letter l} _____________________________________________________
} HASWELL Malcolm wrote: } } } } I want to supply Karnovsky (modified to 2.5% G and 2.5% "paraformaldehyde") } } to a lab to collect 5mm biopsies over a period of time. } } In the past I have used Karnovsky for up to a week after producing it but I } } am not sure about storing it for 1-2 months. Should I store at 4 deg C or } } consider freezing some (it will be in caodylate with 2.5mM CaCl2)? } } } } Malcolm Haswell } } Electron Microscopy } } University of Sunderland } } Malcolm: } } I have stored this fixative in glass at room temperature for extended } periods (1-2 months) and have had no problems. Once you see some bottom } ppt. or cloudiness, discard with copious amounts of water. Also be } careful of the formalin conc. in the air. Best regards, Jerome.
I would recommend purging the vials with nitrogen gas prior to sealing. Use teflon lined caps, if possible and store at 4C.
cheers
Edward J. Basgall, PhD The Pennsylvania State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618 http://www.personal.psu.edu/ejb11/
} } We will be demoing a Noesis pacckage soon. } } Superb but not particularly friendly. Steep learning curve.
Kalman, my records indicate you are using our old version Visilog4.1.3. The new release is much more powerfull and easier to use. Give me a call and I'll upgrade you at no cost.
Regards,
---------------------------------------------------------------------------- -------- Luc Nocente Tel: 514 345 1400 Noesis Vision Inc. Fax: 514 345 1575 e-mail: ln-at-noesisvision.com http://www.noesisvision.com 6800 Cote de Liesse, Suite 200 St-Laurent, PQ H4T 2A7,Canada ---------------------------------------------------------------------------- --------
NANCY SMITH wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } I would be interested to hear from Philips XL30 & XL40 users } regarding how long their tungsten filaments last. } } Thanks for your input. } Nancy R. Smith } Director of Operations } Microscope And Graphical Imaging Center } California State University, Hayward } http://www.csuhayward.edu/SCI/sem
Nancy,
Although we do not use a Philips XL30 or XL40 in our own lab, we have supplied filaments for years. So based on our experience and comments from our customers, let me make the following observations:
Some Factors Affecting Tungsten Filament Life
1) quality of vacuum 2) single or multiple user instrument 3) high or low KV 4) height setting of filament 5) oversaturation 6) quality of the Tungsten
Some of our customers report life spans of 50 to 200 hours and beyond. A normal burnout is when the filament has a bubble on the top of the broken wire. Look for these indicators when the filament burns out too quickly:
a) normal burnout where the base is clean could mean there is a good vacuum but an incorrect height setting or oversaturation. b) a discolered base could mean a bad vacuum c) a cracked filament (no bubble) could be a flaw in the wire or a slight crack in the wire during installation
line resolution of film: films were compared by Downing and Grano (1982) Ultra- microscopy 7:381-404 the Kodak 4463, as used in this study, is now the SO163, as given in a data sheet No P-252 by Kodak. There is no simple figure for the resolution, but they measured the modulation transfer function for different frequencies. Check the paper, it is worth reading. Regards, Reinhard
I'm translating a microscope manual from Japanese into English and constantly r unning into sentences that go like "part of the field of view may be cut off" a nd "image cut-off in peripheral part may occur."
Could anyone tell me how to put them correctly? Should I instead say "the field of view may be vignetted" and "image vignetting may occur" which sound more te chnical to me but I'm not sure about.
Any suggestion is welcomed. Thanks in advance.
Chiba Atsushi [(Mr.) -- *Chiba* is my surname] Voice: (+81) 010-045-9451
Chiba Atsushi wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Sorry for my layman's question. } } I'm translating a microscope manual from Japanese into English and constantly running into sentences that go like "part of the field of view may be cut off" an } } Could anyone tell me how to put them correctly? Should I instead say "the field of view may be vignetted" and "image vignetting may occur" which sound more tec } } Any suggestion is welcomed. Thanks in advance. } } Chiba Atsushi [(Mr.) -- *Chiba* is my surname] } Voice: (+81) 010-045-9451Chiba, I would go with the simpler language. While vignetting is the appropriate term, it is less explicit.
MARK YOUR CALENDARS NOW!!! EVERYONE IS WELCOME TO ATTEND!
The Fall meeting of the Midwest Microscopy and Microanalysis Society will be held on Friday, November 7, 1997, at Eli Lilly & Co. in Greenfield, Indiana (near Indianapolis). The meeting is hosted by Jeffrey Horn of the Toxicology Research Laboratories at Lilly.
The schedule for the meeting is:
8:30 Welcome and Brief Overview of EM Lab Functions at Lilly - J. Horn, M.S., Eli Lilly and Company
9:00 Electron Microscopy in Pharmaceutical Drug Development - V. Meador, DVM/PhD, Eli Lilly and Company
9:30 Everything You Always Wanted to Know About Molecular Morphology (But Were Afraid to Ask) - the Basics of In Situ Labeling Methods - J. Fagerland, Ph.D., Abbott Laboratories
10:00 Coffee Break
10:30 Ultrastructural Evidence in Legal Cases, N. Cheville, DVM/PhD, Iowa State University
11:30 Lunch (Provided)
12:45 Tour Eli Lilly & Co. Toxicology Facility
2:00 Use of SEM for Selecting Therapeutic Candidate in Various in vivo Thrombosis Models - G. Sandusky, DVM/PhD Indiana University
2:30 Diagnostic Electron Microscopy in the Hospital Setting - M. Goheen, M.S. Indiana University Hospitals
3:00 Coffee Break
3:30 Digital Imaging for Dummies - J. Gagne, M.S., Abbott Laboratories
4:15 Conclusion
Meeting announcements, maps, and registration instructions will be sent to members of MMMS in U.S. Mail soon. Anyone else can receive them by contacting me at (847) 935-0104 or via email at jane.a.fagerland-at-abbott.com.
Sanford, first are you speaking of the differences with objectives as seen through eyepieces or with the CCD camera? if the problem is with the CCD camera one problem is probably IR leakage onto CCD with one objective and not the other. The NA of the lens should be taken as a measure of resolution capability not amount of light transmission, though with two objectivesof exact same type of glass and design then the NA can be used to see who collects more light. But when you start to compare a PlanApo to PlanNeofluar then the glass and lens design insdie objective may have drastically different spectral properties, especially outside visible range (Up in IR } 850nm) this is what will cause a low contrast soft image.
Another likely possiblity is that the lower the NA the more depth of field the objective will have and thereby when looking at a 3-d object have a sharper edge as the fluorecesence is coming from above and below focal plane (This is why confocal looks so good and why most quality microscopes will offer, and you should be purchasing an objective with an iris diaphram -- or have body of scope that has it built into body--aperture diaphramfor epi path-- to cut down on NA to cut down on bloom in sample. (your flourescent cell)
To check yourself put both objective on upright scope with high resolution condenser and look at a very thin {5micoron sectionsof H&E stain tissue. at this point the higher NA should be clearer and sharper then the 1.25NA . But other than BF you can see that nore NA is not always better.
Scott E. Berman Advanced Imaging Concepts, Inc. (609) 921-3629 x26 Princeton, NJ scotte57-at-aol.com
Sanford, first are you speaking of the differences with objectives as seen through eyepieces or with the CCD camera? if the problem is with the CCD camera one problem is probably IR leakage onto CCD with one objective and not the other. The NA of the lens should be taken as a measure of resolution capability not amount of light transmission, though with two objectivesof exact same type of glass and design then the NA can be used to see who collects more light. But when you start to compare a PlanApo to PlanNeofluar then the glass and lens design insdie objective may have drastically different spectral properties, especially outside visible range (Up in IR } 850nm) this is what will cause a low contrast soft image.
Another likely possiblity is that the lower the NA the more depth of field the objective will have and thereby when looking at a 3-d object have a sharper edge as the fluorecesence is coming from above and below focal plane (This is why confocal looks so good and why most quality microscopes will offer, and you should be purchasing an objective with an iris diaphram -- or have body of scope that has it built into body--aperture diaphramfor epi path-- to cut down on NA to cut down on bloom in sample. (your flourescent cell)
To check yourself put both objective on upright scope with high resolution condenser and look at a very thin {5micoron sectionsof H&E stain tissue. at this point the higher NA should be clearer and sharper then the 1.25NA . But other than BF you can see that nore NA is not always better.
Scott E. Berman Advanced Imaging Concepts, Inc. (609) 921-3629 x26 Princeton, NJ scotte57-at-aol.com
Carlos E. Barbosa wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Does someone can say me what type of lamp have I to use for performing } UV micropscopy of thin-sections of rocks. } Thank you in advance! } } Carlos BarbosaDear Carlos,
Both high pressure mercury arcs and xenon arcs show good emission in the UV. One caution: most conventional optics do not transmit below about 365-380 nm. Talk to your microscope vendor about quartz optics for all relevant components. I assume that you will be doing fluorescence, since humans cannot see into the UV (although I am told that those of us who have had cataract operations see further into the near UV than those of us with convention eyes) *and* of course, direct viewing of UV light can literally fry the delicate tissue in the eye. With UV fluorescence from a thin section, the job is a bit easier, but you will need quartz optics throughout the illumination system , including the objective. You may also want to test the glue which is holding your thin section to its substrate, to make sure that it is not autofluorescent, creating undesirable background.
Let me know how things work out.
Barbara Foster President Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108 PH: (413)746-6931 FX: (413)746-9311 email:mme-at-map.com --------------------------------------------------------------------------------------------------------------------------------- ********** Microscopy/Microscopy Education ********** America’s First National Consortium of Microscopy Experts Specializing in Customized, On-site Training in all areas of Microscopy, Sample Prep, and Image Analysis
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas, As I understand for this type of staining, it depends on whether the grids are 'formvar' coated or not. If the grids that you are using are coated in some way, even if you did immerse them into solutions, the sections will only be stained on one side anyway. So there is no advantage in immersing the grids.
However, if the sections are on uncoated grids, then immersion into the Ab sols will give you greater staining, (both sides!)
Hope this helps,
Rich.
----------------------------------------------------------------------- Richard Lander Electron Microscope Technician South Campus Electron Microscope Unit Otago School of Medical Sciences P.O. Box 913 Dunedin New Zealand. Tel. National 03 479 7301 Fax. National 03 479 7254
"Southernmost EM Unit in the World!" ------------------------------------------------------------------------
=46or both immuno staining and U acetate/Pb citrate, I've always immersed. In 25 =B5l drops for the former and in a multigrid staining thing for the latter. THe immersion for immuno was to pick up as much stain as possible on a structure that was about half as thick as an EM section. Oh, I was using uncoated grids, of course.
Rosemary White Department of Ecology and Evolutionary Biology Monash University, Melbourne, Victoria 3168, Australia phone 61-3-9905 5670 email r.g.white-at-sci.monash.edu.au fax 61-3-9905 5613 or rgwhite-at-vaxc.cc.monash.edu.au
For reasons best known to ourselves, we are attempting to resurrect an OMAR SPC 900/EX critical Point Dryer. It has not been used in some time and none of us can recall using it, let alone how it was used!! Of course, the manuals have long since disappeared (presumably to the same place that odd socks and pens go) and we need to get it going. Does anybody out there have a manual that they could fax us or even an idea of the pressures and times we should be using with this little beast? Any help would be gratefully accepted!
Many thanks,
Colin V.
Colin J. Veitch Instrumentation Scientist CSIRO Division of Wool Technology PO Box 21, BELMONT, Vic. 3216. Australia.
I work with a LaB6 Auger in a university environment and have been given a project to identify contaminants on and just below the surface of a leadframe (Au on epoxy). I can overcome the charging prior to depth profiling by dropping to low voltage (2Kv or Less). Once I start ion sputtering for short times the charging is enormous and continuous. Any suggestions would be appreciated.
Sincerely,
Tamara E. Bloomer Assistant Scientist Ames Laboratory 137 Wilhelm Hall Ames, IA 50011 (515) 294-2564
The text "Advanced Scanning Electron Microscopy and X-Ray Microanalysis" by Newbury, Joy, Echlin, Fiori, and Goldstein, 1986, Plenum, has a chapter on characterization of semiconductors which covers EBIC and voltage contrast.
regards,
Dave Audette Olin Research Center Cheshire, CT 06410 USA (203)- 271-4272 deaudette-at-corp.olin.com
colin.veitch-at-dwt.csiro.au wrote: } } Hi All, } } For reasons best known to ourselves, we are attempting to resurrect an } OMAR SPC 900/EX critical Point Dryer. It has not been used in some time } and none of us can recall using it, let alone how it was used!! Of } course, the manuals have long since disappeared (presumably to the same } place that odd socks and pens go) and we need to get it going. Does } anybody out there have a manual that they could fax us or even an idea } of the pressures and times we should be using with this little beast? } Any help would be gratefully accepted! } Hi Colin: Times have to be determined by trial and error. It depends upon the sample. As to temperature, the chamber must be cooled to between 15-20C in order for there to be liquid CO2. The solvent used to dehydrate the sample must be removed and this is done with repeated flushing of the chamber. When this is done the temp is raised to above 32C(36C just to be sure) pressure will be around 1200lbs. At this point the critical point has been reached and passed and pressure can be slowly lowered. Another option is Hexamethlydisilazane(HMDS). This is a liquid that can be used in place of CPD. After dehydration HMDS is subistuted with several changes and then allowed to air dry. It gives good results. You will have to try it on your sample
Hope this helps. Good luck. THE OPINIONS HERE ARE NOT THOES OF THE FACILITY Greg Rudomen Greg-at-umic.sunysb.edu University Microscopy Imaging Center S.U.N.Y. Stony Brook Stoy Brook, NY 11794-8088
I think it is O.K. to follow phosphate with cacodylate. We do so routinely, and don't seem to have any problems. Lesley Weston.
On Thu, 11 Sep 1997, John Chandler wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Thanks to all who responded to my osmium pepper question. I now know not } } to osmicate in phosphate but haven't decided what I should use for a } } buffer-I don't think phosphate and cacodylate are compatible so a simple } } switch may be out. Has anyone out there used Dalton's dichromate/osmium? I } } cannot find any reference to anything by Dalton. I do have a formula and } } have used it but am curious to know if anyone else has played with it. Tx } } Grace } } In a former life, I used Dalton's fixative for immersion fixation of } vertebrate retina. I don't remember all the details, but the results were } very good. I don't remember this fix being widely used, though. } } John } chandler-at-lamar.ColoState.EDU } } }
I need a TPR 010 Piranni gauge for my Balzers BAF 301 Freeze etcher. Techno Trade does have them for $250.00 each...which is ok on an unlimited budget. Has anyone else gotten these elsewhere? Is there an aftermarket brand that I might use? And yes I have cleaned many a TPR 010 {I have a real good technique...} but after around four cleanings they act very strange. Any help, advise, or berations are welcome.
John Grazul Rutgers University Electron Imaging Facility
All manuals should follow the "KISS" principle! ie: Keep It Stupidly Simple!!!! Forget about whether it sounds "technical" or not, just make sure the directions are abundantly clear to a 5 year old child and researchers around the world will applaud you.
} } Sorry for my layman's question. } } I'm translating a microscope manual from Japanese into English and } constantly running into sentences that go like "part of the field of view } may be cut off" and "image cut-off in peripheral part may occur." } } Could anyone tell me how to put them correctly? Should I instead say "the } field of view may be vignetted" and "image vignetting may occur" which } sound more technical to me but I'm not sure about. } } Any suggestion is welcomed. Thanks in advance. } } } Chiba Atsushi [(Mr.) -- *Chiba* is my surname] } Voice: (+81) 010-045-9451
Cheers :) George Department of Earth and Atmospheric Sciences University of Alberta Edmonton, Alberta T6G 2E3 Canada ph: 403-492-5746 fax: 403-492-2030
Anyone aware of a "nearly" conventional photographic process for development of prints made from enlarged negatives? Examples: stabilization processors that use heat to develop sensitized papers OR (less desirably) concentrated chems as in the Kodak Ektamatic-like scheme.
Unfortunately, the heat-developed papers use mercury and silver and will be phased out shortly while the Ektamatic-type stabilization processors generate liquid wastes. The ideal situation would involve conventional negs enlarged onto a paper that could be processed using some sort of dry (or nearly so process) AND yielding photographic quality images.
Now, digital imaging people please don't jump all over me, because sometimes digital images and prints just do not offer adequate rendition of the information -- without spending big bucks. Right now, we are working with dense, highly contrasted negatives (difraction patterns, high contrast and thick specimens) and the quality of the digital images (direct image capture or scanning of TEM negs) is inadequate. My own idea would be to have a system akin to a xerographic process wherein one would enlarge a negative onto a high quality paper that would then be "developed" in a manner similar to a dry copier probably using toner powders. I believe all of these components currently exist but have not been put together - to my knowledge.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
O.k., I know this has been discussed MANY times before, but I I didn't save all the relavent messages.
Looking for two things:
(1) Refurbished / replacement screen for a JEOL 100-S (hey it still works fine)
(2) Looking for places to recoat our old 100s screen. I believe that SPI still does it, and will call soon (I did save your message Chuck). Grant Sci Corp doesn't seem to exist any more. Any others to consider? Any comments god or bad?
Thanks.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"640K ought to be enough for anybody." -- Bill Gates, 1981
We have 10 years of blocks filed and stored in small plastic boxes from Althor Products. Our last purchase order was returned: no forwarding address. Does anyone know how to contact these people or of another source for those 1 3/4 x 7/8 x 3/4" boxes with snap-on lids? Thanks Joyce Craig Chicago State University
We inherited an old embedding oven that is caked in resin. Does anyone ever clean out their embedding oven? If so, what is used to do this?
I am not so concerned about the cleanliness of the oven, but rather the problems it seems to be causing when things start sticking to the oven. There is a plastic petri dish with old desiccant in it that has stuck itself onto the bottom of the oven, with no way to easily dislodge it.
Any suggestions or comments would be greatly appreciated.
Susan
Susan Carbyn Atlantic Food and Horticulture Research Centre Agriculture and Agri-Food Canada Kentville, Nova Scotia B4N 1J5 Canada
I am making TEM samples of modern and fossil brachiopod (a bivalved marine invertebrate) shells composed of calcite. I was intending to use an ion milling device (argon) to thin the samples. However, it has been suggested that a tripod polisher would be less time consuming and provide a larger "viewing" area.
Has anyone had experience using a tripod polisher on carbonate materials? Would tripod polishing produce more defects in the crystallographic structure than ion thining? My intent, if possible, is to evaluate diagenetic alteration by observing crystallographic defects.
Thanks.
Nancy Buening
Nancy Buening E-mail: buening-at-geology.ucdavis.edu Department of Geology Phone: (916) 752-0350 University of California Fax: 916 752-0951 Davis, CA 95616
Julian, I didn't expect the reply so quick. I certainly try that. Thanks so much. Regarding serial sectioning, you didn't mention what kind of knife you used for GMA sectioning. Is tungsten or glass knife? Which is better? Regular Oil Red O staining procedure won't work with GMA section. Why? The reason we choosing GMA is that the morphology is better than frozen tissue. Do you have suggestion about this? Dorothy
We recoat screens in our lab. Please contact us for pricing.
Dianne Ernest F. Fullam, Inc.
O.k., I know this has been discussed MANY times before, but I I } didn't save all the relavent messages. } } Looking for two things: } } (1) Refurbished / replacement screen for a JEOL 100-S (hey it still } works fine) } } (2) Looking for places to recoat our old 100s screen. I believe } that SPI still does it, and will call soon (I did save your message } Chuck). Grant Sci Corp doesn't seem to exist any more. Any others } to consider? Any comments god or bad? } } Thanks. } } } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 352 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } } } "640K ought to be enough for anybody." } -- Bill Gates, 1981 } } Ernest F. Fullam, Inc. Phone: (518) 785-5533 FAX: (518) 785-8647 E-Mail: pdf-at-fullam.com
I apologize to Grant Scientific Corp for passing apparent misinformation, for indeed Grant Scientific does exist (as it has since 1975 I have been informed). I have spoken with the good folks at Grant, and they have been able to provide me some very useful information regarding TEM Screen re-coating.
I wish to make it all known to the listserver goupr that Grant does exist and can be contacted at: 803-829-2841.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"640K ought to be enough for anybody." -- Bill Gates, 1981
I am replying to my subscription request. All information is correct.
Nancy Buening
Nancy Buening E-mail: buening-at-geology.ucdavis.edu Department of Geology Phone: (916) 752-0350 University of California Fax: 916 752-0951 Davis, CA 95616
} We inherited an old embedding oven that is caked in resin. Does anyone } ever clean out their embedding oven? If so, what is used to do this?
I used to chisel away at the stuff, and try to use various solvents to clean off the glass door without dissolving my gloves, with limited success. One day I came into the lab and, lo! the oven was clean. Turns out someone had cleaned it out with our putty knife WHILE IT WAS HOT! Duh. She said it was pretty easy. Be careful of all surfaces. Wear gloves. Worry about toxicity. Disregard me if anyone says this is not recommended.
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
} } We inherited an old embedding oven that is caked in resin. Does anyone } ever clean out their embedding oven? If so, what is used to do this? }
I am not sure what kind of resin is involved, but once I had to clean out a 5 litre three-neck flask that had been used to synthesize alkyd resins - these are basically phthalic polyesters crosslinked through polyunsaturated fatty acids. There was a caked-on film that would not yield to concentrated sulphuric acid, tetrahydrofuran, or any of the usual things. I put some .880 Ammonia in the bottom of the flask, blocked the necks with cotton wool, and left it overnight, and next morning the film had swollen and fallen away and could be yanked out with tongs or whatever.
Ammonia vapour is very effective with polyester based resins because of (a) it basic nature and (b) most important, its small molar volume.
If, on the other hand, your resin is an epoxy, it might be better to put a dish of methylene chloride (dichloromethane) in the bottom, seal the oven, and go away overnight. Methylene chloride is the basis of most commercial paint strippers.
The use of vapour technique does make for much less messy operation. Once the film is loosened, strong detergent should be good enough for scrubbing.
However, that PLASTIC PETRI DISH would probably turn into a gooey mess with the methylene chloride vapour, so try the ammonia first. (Also consider, does your oven seal with an O-ring?)
+------------------------------------------------------------------------+ | 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 have a procedure for Oil Red O staining of GMA sections which was sent to me by Bob Schoonhoven. I haven't tried it myself.
Stain: Oil Red O Indication: Demonstrate lipids Solutions: 1) 60% aqueous triethyl phosphate 2) 0.5% Oil Red O solution 0.5g Oil Red O (CI 26125) 100 ml 60% aqueous triethyl phosphate Filter before use 3) Celestin Blue 0.5g celestin blue B 100 ml 5% aqueous ferric ammonium sulfate Boil gently 2-3 minutes; cool to room temperature; filter; and add 12 ml glycerol Filter before use Procedure: 1) Rinse briefly in 60% triethyl phosphate 2) Stain 5-20 minutes in Oil Red O solution 3) Rinse 1-2 seconds in 60% triethyl phosphate 4) Rinse well in distilled water 5) Counterstain in Celestin Blue 15 minutes 6) Rinse well in distilled water 7) Mount in glycerin jelly or other water-soluble mount Results: lipids: red-orange nuclei: blue Reference: Feldman, A.T. and Dapson, R.W., "Relative Effectiveness of Various Solvents for Oil Red O," _Medical Laboratory Technology_, Vol. 31:335-341, 1974.
Disclaimer: Energy Beam Sciences manufactures the JB-4 and JB-4A microtomes for sevtioning plastic-embedded tissue, and sells GMA kits.
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
The vignetting I most frequently see is a cut-off of the image at the corners due to the microscope's camera tube. This can commonly be caused by using an adapter (for example, a C-mount adapter for the digital or video microscope camera) with too wide a field of view. The descriptions you cite in the Japanese text could be describing this or other phenomena. You'd need to know more from the original author(s) to be sure. Brooks Corl Senior Applications Manager, Polaroid Corporation E-mail: corlb-at-polaroid.com
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
In a message dated 97-09-12 08:45:19 EDT, Chiba writes: { { sentences that go like "part of the field of view may be cut off" and "image cut-off in peripheral part may occur." Could anyone tell me how to put them correctly? Should I instead say "the field of view may be vignetted" and "image vignetting may occur" which sound more technical to me but I'm not sure about. } }
The phrase "part of the field of view may be cut off" sounds ok to me. Without an illustration, I do not know what it means. My mental image is that there is a black part within a photographic image taken with the microscope from a flash/shutter timing problem, or from blocking of the field of view by an accessory that is inserted in the optical path, such as the quarter wavelength plate typically used in a polarizing microscope.
The phrase "image cut-off in peripheral part may occur." could be reworded as "...the peripheral part of the image may be cut-off." As above, I do no know really what it means. But I imagine that it means the effect from closing down the sub-stage iris. I do not think that is really vingetting, a gradual shading, but it is close to it. If the effect being discussed is from gross misalignment of the bulb in the illuminator housing, then it could be vignetting.
Steve Stokowski Stone Products Consultants Concrete Petrographers http://members.aol.com/CrushStone/index.htm ---------------------------------- Forwarded ----------------------------------
Are you into digital? Please help to reevaluate digital image processing for the development of a program for next year's MSA-98 meeting (Follow-up and Summary will be placed here):
"Applied Image Processing: What Can It Do For Digital Imaging"
If you like to help, type your reply directly into this message and send it to me at
Klaus-Ruediger Peters {Peters-at-bsac.uchc.edu}
Thanks for your help and voicing your opinion. Klaus ****************************************************************************
What is important, what are you using, what deserves more attention, what has "practical value" and may represent one of the topics? What is of your interest? What can't you do but like to? Please, hit "REPLY" now, then "PASTE" my address into "TO:" and just fill in your Spontaneous thoughts on:
-----------------------------------Being Digital--------------------------- Digital image handling (Shifting data, formatting, labeling, annotating, 8-bit to 16-bit/per channel)
----------------------------------Trying Digital----------------------------- Digital darkroom (Printing images as hard copies on paper, overheads and slides)
----------------------------------Doing Digital----------------------------- Digital image display and "enhancement" (Presenting the desired information)
---------------------------------Using Digital------------------------------ Data quantification (Finding and measuring the desired information)
----------------------------Other topic of interest?------------------------
Images are as diverse as their usages. But, what are the common concepts and how good do they apply to the common daily imaging tasks? Any possible contributions from yourself to this MSA 1998 program?
Thanks for your initiative and for your help. Klaus
The Minnesota Microscopy Society FALL BUFFET DINNER & TALK will take place Tommorrow, SEPTEMBER 18, 1997 from 5:30 - 8:00 PM at The Campus Club, University of Minnesota Minneapolis, East Bank Campus
SPEAKER: William P. Wergin Agricultural Research Service,U.S. Dept. of Agriculture, Beltsville, MD
TOPIC: "THE MICROSCOPY OF SNOW:" "The 3-D Structure and Metamorphoses of Snow and Ice Crystals as Revealed by Low Temperature SEM". For more details see our website at http://resolution.umn.edu/MMS/
We hope to provide a pleasant evening during which microscopists will be moved to renew or begin memberships in MMS, MSA and/or MAS.
Program 5:30-6:00 Wine, Cider & Cheese Social 6:00-7:00 Buffet Dinner. 7:00-8:00 Talk The Buffet Dinner is $12 per MMS member, payable at the door. (non-member fee is $22, includes new membership, to attend without becoming member, $15.) STUDENTS: NEW FEATURE THIS YEAR: Current student members or new student members - $5.00 membership fee payable at the door- will receive a complimentary buffet dinner courtesy of MMS and sponsoring vendors.
Please make an advance reservation by contacting:
Mike Coscio (612)569-1331, 569-1284 FAX, mike.coscio-at-medtronic.com Stuart McKernan, (612)626-7942, 626-7530 FAX, stuartm-at-maroon.tc.umn.edu
Parking is available behind the Union in the East River Road Ramp (connected by walkway to Union) for $2.50 (per day rate), at the Radisson Ramp on Washington Ave. S.E., a block east of the Union, and at other Minneapolis Campus locations
__________________ Stuart McKernan stuartm-at-tc.umn.edu Microscopy Specialist CIE Characterization Facility, University of Minnesota Phone: (612) 626-7594 100 Union Street S. E., Minneapolis, MN 55455 Lab: (612) 624-6590
A collegue contacted me asking for information on labeling of a small protein in potato tuber. Apparently, she is having problems with the potato starch disintegrating in the electron beam. Do any of you have any suggestions about how to retain antigenicity and, at the same time, attain good (or reasonable) fixation. If so, please contact her offline at the email address below as she is not yet a member of this list.
Her name is Bonnie Compas: email address is: becompas-at-csupomona.edu
Thank you,
De Wood
*****************************************
Delilah F. Wood United States Department of Agriculture Western Regional Research Center 800 Buchanan Street Albany, CA 94710
} Did anybody know the size of large and small viewing screen of CM120 } and CM200/FEG. Your information will be greatly appreciated! } gary } } } +--------------------------------------------------------------------+ } | Gary Gang REN, Ph.D. | } | Department of Cell Biology #MB219 Email: gangren-at-scripps.edu | } | Mail Stop MB21 Tel: (619) 784 9815 (O) | } | The Scripps Research Institute (619) 546 1585 (H) | } | 10550 N. Torrey Pines Road Fax: (619) 784 9927 | } | La Jolla, CA 92037, USA http://leonardo.scripps.edu/ren | } +--------------------------------------------------------------------+ } } }
I am looking for a person to fill a 1 year surface chemist position at a petroleum company in Beacon, NY (Dutchess County). The person must have Ultra High Vacuum, X-Ray Spectoscopy (XPS) and Scanning Electron Microscopy (SEM)or AES experience. The chemist will be doing sample prep and intro, analysis, and running data. It is an applied research, product related position. Please e-mail me at kwagscha-at-aerotek.com or call 1-800-973-1518 ext. 1045 if interested. Kristen Wagschall
I am posting this for a friend who is not on the listserver. Please reply=
directly to Trinity College.
Thank you!
David Henriks South Bay Technology, Inc.
************************ PLEASE POST ************************
Department of Human Resources Trinity College Hartford, Connecticut 06106
Position Announcement
Electron Microscopy Laboratory Manager/ Technician
Trinity College has an immediate opening for an experienced electron microscopist to work in a newly established Electron Microscopy Center. A= s this facility is designed to serve both the physical and biological sciences, familiarity with both disciplines is highly preferred. Primary responsibilities include assisting faculty and students in the use of the=
facility for teaching and research, and overseeing the upkeep and use of TEM, STEM and specimen preparation facilities, including the analytical E= M EDX and EELS). A B.S. degree with advanced research training and/or extensive experience with TEM are essential; master's degree with requisi= te experience is preferred.
This position is a full-time, 12-month appointment with full College benefits. Applications will be reviewed upon receipt; search will continu= e until position is filled. Please respond with a resume, cover letter stating salary expectatons, and the names, telephone numbers and addresses of three professional references to: Prof. Daniel Blackburn, c= /o Human Resources, 300 Summit Street, Trinity College, Hartford, CT 06106.=
(Resumes also may be faxed: (860) 297-5140, or sent via the internet: Sandra.Magee-at-Mail.Trincoll.edu).
Trinity College is an Equal Opportunity/Affirmative Action Employer. Wome= n and mminorities are encouraged to apply. Applicants with disabilities should request any needed accommodation in order to participate in the application process.
The Duniway Stockroom Corp. ((800-446-8811) deals in new, used, and rebuilt vacuum devices, including all types of vacuum gauges. They might be able to help you with a replacement gauge, or possibly they could rebuild yours.
Good luck,
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
I have received a couple of inquiries about what Torr Seal is and where to get it.
Torr Seasl is an epoxy compound especially formulated for use in vacuum systems that was introduced by Varian Associates, Vacuum Products Division, 121 Hartwell Ave, Lexington, MA 02173-3133 (800-882-7426) a number of years ago. It is stated to contain no solvents, and therefore to be good at pressures below 10-9Torr. It is bakeable at temperarures up to 120C. It adheres to most clean materials (glass, metals, ceramics), and holds up well over long term service. Some companies that handle EM supplies also handle it (e.g. I find it listed in the Ladd catalog, probably SPI handles it too).
Other similar products are also sold by other companies. For example, Duniway Stockroom Corp. (800-446-8811; info-at-duniway.com) sells a product called 'Epoxy Patch' that is stated to be equivalent to Torr seal
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
Dear Microscopists, I am forwarding this message from SAFETY listserver. Marek Malecki.
On Sep 17, 8:56am, Dr. Gary Gang REN wrote: } Subject: screen size } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Did anybody know the size of large and small viewing screen of CM120 } } and CM200/FEG. Your information will be greatly appreciated! } } gary } } } } } } +--------------------------------------------------------------------+ } } | Gary Gang REN, Ph.D. | } } | Department of Cell Biology #MB219 Email: gangren-at-scripps.edu | } } | Mail Stop MB21 Tel: (619) 784 9815 (O) | } } | The Scripps Research Institute (619) 546 1585 (H) | } } | 10550 N. Torrey Pines Road Fax: (619) 784 9927 | } } | La Jolla, CA 92037, USA http://leonardo.scripps.edu/ren | } } +--------------------------------------------------------------------+ } } } } } } } } } -- End of excerpt from Dr. Gary Gang REN
You can measure it quite accurately by using the measuring function of the CM scope. Just move any thing you can identify from one side of the screen to the other side of the screen.
Yifan Cheng
-- ********************************************************************** * Dr. Yifan Cheng * Phone: +1-850-644-4104 (Office)* * Institute of Molecular Biophysics * or: +1-850-644-9769 (Lab) * * Florida State University * Fax: +1-850-561-1406 * * Tallahassee, FL 32306-4380 * Email: ycheng-at-sb.fsu.edu * * U.S.A. * http://www.sb.fsu.edu/~ycheng * **********************************************************************
Dear Microscopists, To evaluate effects of various cryoprotectants on the cells' viability, we would like to image cultured cells being frozen. Is anybody aware of a cryo-stage (below -100deg.C) for light microscopy? Any information will be greatly appreciated. Vendors welcome. Sincerely, Marek Malecki.
FOR SALE: Brand New 6.2 mm Dupont diamond knife. Never used. Current retail for a knife of this length is approximately $6000+. I would like to sell the knife for $3200 or best offer. Ron Kalil
Ronald Kalil Center for Neuroscience University of Wisconsin 1300 University Ave. Madison, WI 53706
Has anyone ever heard of something called the "Arkograf electrical metal etching pen?" Or more importantly, the name of the manufacturer and where they are located? It is a device for making permanent markings on metal surfaces for identification purposes.
Thanks.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
We have an Electric Discharge Machine Mark III, Ser 338854; It came originally from Concept EDM Ltd, Maidenhead, Berkshire England. I checked the UK yellow pages but they seem no longer to exist.
Our problem is that the Electric Discharge Machine won't work and we would like the circuit diagrams (schematics) before trying to fix it.
If anyone out there has knowledge of the current address etc. of the original suppliers, OR has a circuit diagram they could copy for us, we would be most grateful.
Thanks,
Mel Dickson President, Australian Society for Electron Microscopy Director, Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
My belated thanks for responding to my query into "vignetting" I posted on the Microscopy list. Your comment was very helpful and enable me to get the job done.
Thank you very much again.
Chiba Atsushi [(Mr.) -- *Chiba* is my surname] Voice: (+81) 010-045-9451
My belated thanks for responding to my query into "vignetting" I posted on the Microscopy list. Your comment was very helpful and enable me to get the job done.
Thank you very much again.
Chiba Atsushi [(Mr.) -- *Chiba* is my surname] Voice: (+81) 010-045-9451
My belated thanks for responding to my query into "vignetting" I posted on the Microscopy list. Your comment was very helpful and enable me to get the job done.
Thank you very much again.
Chiba Atsushi [(Mr.) -- *Chiba* is my surname] Voice: (+81) 010-045-9451
My belated thanks for responding to my query into "vignetting" I posted on the Microscopy list. Your comment was very helpful and enable me to get the job done.
Thank you very much again.
Chiba Atsushi [(Mr.) -- *Chiba* is my surname] Voice: (+81) 010-045-9451
} Dear Microscopists, } I am forwarding this message from SAFETY listserver. } Marek Malecki. } } Date: Wed, 17 Sep 1997 13:25:29 -0400 } From: "Ralph Stuart, Vermont SIRI" {rstuart-at-esf.uvm.edu} } } A Tiny Drop Of Mercury Shatters Lives And Science } } A9 1997 The Associated Press } } LYME, N.H. (September 13, 1997 6:45 p.m. EDT) - } snips
} "She loved her work," he says. "It made her happy." } } She couldn't have known the risks. She couldn't have known how bad the bad } stuff really was. Truth is, no one knew. } } Just a tiny drop of liquid. Sweet-smelling. Dense. Deadly. } } By HELEN O'NEILL, The Associated Press
While not wishing to diminish the dangers of mercury, I really don't see that this sort of journalistic hype helps anybody. It makes a good story, I suppose, for the sort of newspaper that runs 'Elvis Lives' stories, but it isn't science and even the courts regard hearsay as highly unreliable.
} Dear Microscopists, } I am forwarding this message from SAFETY listserver. } Marek Malecki. } } Date: Wed, 17 Sep 1997 13:25:29 -0400 } From: "Ralph Stuart, Vermont SIRI" {rstuart-at-esf.uvm.edu} } } A Tiny Drop Of Mercury Shatters Lives And Science } } A9 1997 The Associated Press } } LYME, N.H. (September 13, 1997 6:45 p.m. EDT) - } snips
} "She loved her work," he says. "It made her happy." } } She couldn't have known the risks. She couldn't have known how bad the bad } stuff really was. Truth is, no one knew. } } Just a tiny drop of liquid. Sweet-smelling. Dense. Deadly. } } By HELEN O'NEILL, The Associated Press
While not wishing to diminish the dangers of mercury, I really don't see that this sort of journalistic hype helps anybody. It makes a good story, I suppose, for the sort of newspaper that runs 'Elvis Lives' stories, but it isn't science and even the courts regard hearsay as highly unreliable.
Terribly embarrassed... Please accept my apology for the 5 or 6 private thank-yous I just posted. I just kept hitting reply without realizing it was a mailing list.
Chiba Atsushi [(Mr.) -- *Chiba* is my surname] Voice: (+81) 010-045-9451
The procedure that Steven posted does indeed work very nicly.....Provided that no alcohol was used in the "dehydration" process. If you wish to look for lipids in GMA embedded tissues you must use a series of graded (with water) monomer solutions instead of alcohols. Somewhere ????? I have the procedure written but I can't put my hands on it right now (ie: I haven't got a clue as to where I 'filed' it). :(
regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
My thoughts exactly, see the August? issue of Scientific American for a better tribute to Karen Wetterhahn and a less subjective discussion of the risks associated with chemical handling.
} } While not wishing to diminish the dangers of mercury, I really don't see } that this sort of journalistic hype helps anybody. It makes a good story, I } suppose, for the sort of newspaper that runs 'Elvis Lives' stories, but it } isn't science and even the courts regard hearsay as highly unreliable. } } Regards, } } Larry Stoter
================= C. John Runions Section of Ecology and Systematics Corson Hall Cornell University Ithaca, New York USA 14853
Evex Analytical, manufacture of the VIDX X-ray Analyzer and the VIDX = Scan Digital Imaging system for SEM/STEM and TEM, would like your feed = back.
We are now engineering Version 3.0 of our VIDX X-ray analyzer software. = We would like to ask you for your wish list. Your wish list may include = functions, routines, macros, etc. which you would like to see = implemented in your ideal x-ray analyzer, This inquiry is open to every = one.
Please respond via e-mail. You may foward drawing also.
Thank you
Peter Tarquinio Evex Analytical www.evex.com 609-252-9192 Tel 609-252-9091 Fax
Linkam make a very nice heating/cooling stage for light micrscopes with a temperature range of -196 to 600 C. They are distributed locally by Fryer Co. Phone 847-669-2000.
Regards,
Joe Neilly Abbott Laboratories Microscopy and Microanalysis 200 Abbott Park Rd. Abbott Park, IL 60064
I am posting this message for a friend who would like to have the e-mail address of Bill McManus, Utah State Univ., Dept . Biology, Logan, Utah. Please reply to :
Kalabm-at-em.agr.ca
Ann Fook Yang EM Unit, ECORC, Agriculture and Agri-Food Canada, Central Experimental Farm, Ottawa, Ontario, Canada K1A 0C6
If anyone is over in UK at the end of February please bear in mind the fol= lowing meeting....
************************************************************* Microscopy of Internal Interfaces: Determination of= Nanochemistry
Institute of Physic= s, 76 Portland Place, London W1N 4AA,= UK.
Friday 27 February 1998=
10.00am - 5.30pm
Organizer: Dr Rik Brydson, School of Process, Environmental and Materials = Engineering, University of Leeds, Leeds LS2 9JT, UK.
EMAG Committee of Institute of Physics Joint with Royal Microscopical Society: Materials Section.
The chemistry of internal interfaces in both structural and functional mat= erials is often a critical factor affecting resultant physical properties, sometimes overrid= ing or controlling the effects of interface structure. The majority of interfaces have a spatial = extent, in at least one dimension, of a few nanometres. Clearly it is desirable to be able to dete= rmine accurately interfacial chemistry and bonding. With this in mind, a one day IOP meetin= g will be held in London at the end of February 1998 on the techniques for interfacial micro= analysis and relevant case studies in this rapidly expanding field. The day will be div= ided into fundmentals and applications sessions.
Confirmed Invited Speakers: oProf. Mick Brown (Cambridge) "Grain boundary chemistry in metals and all= oys" (Sponsored by RMS). o Dr Rik Brydson (Leeds) "Interfacial bonding determined by EELS" o Dr David Jefferson (Cambridge) "Determination of surface chemistry using= HREM" o Prof. John Titchmarsh (Sheffield Hallam) "Determining Interfacial Segreg= ation using EDX" o Dr John Watts (Surrey) "Surface Analysis applied to Interfaces" o Prof Bruce Hamilton/ Dr Uschi Bangert (UMIST) "STM/STEM of cleaved semic= onductor multilayers" o Dr Paul J Warren (Oxford) "Atom Probe Field Ion Microscopy of Internal I= nterfaces".
Other topics will include: o Interfacial Chemistry and HREM o STM on cross-sections o Theoretical aspects of interface chemistry o Metal-support interactions in Supported Catalysts o Semiconductor interfaces o Biomaterial interfaces o Coatings o Corrosion Films
Additional POSTER contributions are extremely welcome.
For further details about registration etc. please contact IOP conference desk Tel: 0171 470 4800 Fax: 0171 470 4848 Email: physics-at-i= op.org or Rik Brydson Tel: 0113 233 2369 Fax: 0113 242 2531 Email: mtlrmdb-at-leeds.ac= .uk _____________________________ Dr. Rik Brydson, University Research Fellow, Electron Optical Unit, Department of Materials, School of Process, Environmental and Materials Engineering University of Leeds, Leeds LS2 9JT, U.K.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
As part of the group involved in the postmortem examination of the death of Karen Wetterhahn I was grateful for all the information and warnings I could lay my hands on before I dealt with any material. The event ,however reported, serves as a reminder to all of us of the dangers of our materials and the need to keep up with the information involved in our safety. This institution has completely reexamined it's glove policy.
Hey Larry- how many electron microscopists have you met who's hand trembles and shakes as they reach out to shake your hand, or dumps the food from their fork or plate as they struggle to eat lunch? I believe they, their family and friends don't look at this as trash journalism, we should all remember SAFETY FIRST. -Mike Rock
On Thu, 18 Sep 1997, Larry Stoter wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Dear Microscopists, } } I am forwarding this message from SAFETY listserver. } } Marek Malecki. } } } } Date: Wed, 17 Sep 1997 13:25:29 -0400 } } From: "Ralph Stuart, Vermont SIRI" {rstuart-at-esf.uvm.edu} } } } } A Tiny Drop Of Mercury Shatters Lives And Science } } } } A9 1997 The Associated Press } } } } LYME, N.H. (September 13, 1997 6:45 p.m. EDT) - } } } snips } } } "She loved her work," he says. "It made her happy." } } } } She couldn't have known the risks. She couldn't have known how bad the bad } } stuff really was. Truth is, no one knew. } } } } Just a tiny drop of liquid. Sweet-smelling. Dense. Deadly. } } } } By HELEN O'NEILL, The Associated Press } } While not wishing to diminish the dangers of mercury, I really don't see } that this sort of journalistic hype helps anybody. It makes a good story, I } suppose, for the sort of newspaper that runs 'Elvis Lives' stories, but it } isn't science and even the courts regard hearsay as highly unreliable. } } Regards, } } Larry Stoter } } }
I have recently bought several different types of bulbs from Lamp Technology (Bohemia, NY) at 800-533-7548. There prices are a lot better that anything I have ever found through a microscope dealer. One of my dealers told me that I was getting a better price than he was wholesale. I recommend you give them a call. They have a web page but I can't remember the address - use a search engine if you are interested. I have no interest in the company except as a very happy user.
} Does anyone know of a source for the HBO 100W/2 100 watt bulbs } used for fluorescence light microscopy? } } Thanks, } } Dennis Shubitowski } University of Michigan } School of Dentistry } dshubito-at-umich.edu
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
They were rated a favorite last year by Beth Richardson (UGA) who was on a similar quest.
(No affiliation blah, blah, blah.....)
cheers ed
Edward J. Basgall, PhD The Pennsylvania State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618 http://www.personal.psu.edu/ejb11/
Advanced MicroBeam, Inc. would like to announce its new web presence to t= he microscopy community. We can be found at http://www.advancedmicrobeam.co= m. Please take a look at how we may be of service to you.
Thanks Dave
=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D=3D= =3D=3D=3D=3D=3D=3D=3D=3D=3D David A. Stanford Advanced MicroBeam, Inc. 4217C King Graves Rd PO Box 610 Vienna OH 44473-0610
-------------------------------------------------------------- Dr. Patrick L. Huddie (301) 725-2775 Fax (301) 725-2941 Microcosm, Inc., 9140 Guilford Road, Suite O, Columbia, MD 21046 e-mail phuddie-at-microcosm.com URL http://www.microcosm.com The Web is:"Vaster than empires and more slow" Andrew Marvell (1621-1678)
Without in any way downplaying the tragedy for the people involved, the dose-response relationship postulated in the story is not very plausible. Mercury toxicity has been studied extensively for at least half a century. It is a cumulative poison which does cause the type of neurological effects reported, but a "single tiny drop", through a glove and quickly washed off, being lethal seems suspect. Associated Press is not peer reviewed (although maybe it depends on your definition of "peer").
In 1996, the EPA published a 7-volume report on environmental mercury. For some data from recent animal studies (rodent and primate) and the current EPA and FDA reference exposure limits, try the following Web pages.
An excellent sample to test resolution at high tilt angles is a single crystal silicon wafer. If mounted as a cross-sectional sample (electron beam parallel to a {011} zone axis), there are several advantages. In th= is initial orientation, the (111) lattice spacings of 3.14 A, (200) spacings=
of 2.715A and (220) spacings of 1.92A are all available for resolution tests. Being a cubic single crystal, it is a simple procedure to follow t= he Kikuchi bands to any other cubic zone axis for the additional resolution checks you mentioned.The resolution at high tilt angles can be checked by=
mounting the cross-sectional silicon sample with the sample's glue line either parallel or perpendicular to the sample holder rod. With the glue=
line parallel to the sample holder rod, {100} zone axes are accessible by=
rotating the sample holder 45 degrees in either direction. The {100} zone=
includes (220) spacings (200) spacings for resolution tests. Similarly, = if the sample is mounted with the glue line perpendicular to the sample hold= er rod, the second tilt attachment can be used to tilt the sample 45 degrees=
to the {100} zone axis in either direction about the second tilt axis. = If your goniometer has a limited tilt capability, there are many other choce= s of zones at lower (and higher) angles for you to choose from.
While you are performing these resolution tests, there are several other calibrations that are easy to perform and can be done with the same sampl= e. For example, the complete magnification range of the TEM (the supplied values from the manufacturer can be 5-10% off), the camera constant calibration, and the imge/diffraction patter rotation calibration. All o= f these tests and calibrations are extensively documented for the MAG*I*CAL=
calibration sample, which is a single crystal of silicon upon which are precisely grown calibration marks. =
PLEASE NOTE: South Bay Technology, Inc. does offer the MAG*I*CAL Calibration Standard and so we have a finiancial interest in promoting it= s use. For more inforamtion on the MAG*I*CAL, please contact me off-line.
David Henriks TEL: 800-728-2233 (toll-free in USA) South Bay Technology, Inc. 714-492-2600 1120 Via Callejon FAX: 714-492-1499 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. I have a couple of questions and will appreciate any replies and =
suggestions.
1) What is the best sample to test the resolution of a TEM at a tilt angle (} 30degree)?
I am testing the resolution of a CM300FEG microscope. I used oriented gold, evaperated gold particles, graphitized carbon and so on. There is no =
problem of those samples at zero tilt. But after tilt the specimen to 30 degree or =
higher, I got only the good resolution (2.3A in case of evaperated gold particle) =
in one direction, along the tilt axis. I believed it is not because of the =
focus gradient that I could not get the same resolution in both directions at =
this tilt angle. I think the samples (evaporated gold particles and graphitized carbon) may not be the suitable samples for the resolution test of the high tilt angles (higher than 45 degree and up to 60 degree). I wonder if any of you has done such test before and has any suggestions on which specimen should =
be used in this test and where to get it?
2) Is there any published documentation about the line resolution of Kodak SO-163 film?
I am writing an paper about the performance of the CM300FEG at low magnificaiton, and need to know the line resolution of Kodak SO-163 =
(something I can cite for). But I could not find any published documentation. Of =
course, I called Kodak technique support and of course they didn't give me a clue. There is a web site called "Bibliography on EM Imaging and Related Technologies" (http://www.uct.ac.za/depts/emu/imaging/papers.htm), but I can not find any thing there either about SO-163. The only other related information I got =
was from a similar discussion in this board a year ago. But the data about line resolution found in that discussion were mostly estimated but not from any published documentation. I wonder if any of you know there is any kind of published documentation including research paper, technique report or so which gives the line resolution of Kodak SO-163. (BTW, the topic about the line resolution has been discussed a year ago and I am sorry to ask it again.)
I appreciate any information or suggestion on my request.
Yifan Cheng
BTW, I am not sure if my email address is already on the list or not. So that I appreciate also that when you reply this email, send a reply to me directly as well as to the board. Thanks again.
-- ********************************************************************** * Dr. Yifan Cheng * Phone: +1-850-644-4104 (Office)* * Institute of Molecular Biophysics * or: +1-850-644-9769 (Lab) * * Florida State University * Fax: +1-850-561-1406 * * Tallahassee, FL 32306-4380 * Email: ycheng-at-sb.fsu.edu * * U.S.A. * http://www.sb.fsu.edu/~ycheng * ********************************************************************** =
Dear reader, I tried to detect mRNA and protien in the same slide. My insitu hybridyzation and immunohistochemistry both worked. but not working when I put them together. I tried insitu first then IHC. Is there anyone tell me some tricks about it? Thanks. Dorothy
Kate, would you share with us the outcome of your institution's re-examination of its glove policy? Thanks. /Vachik Hacopian
} As part of the group involved in the postmortem examination of the death of } Karen Wetterhahn I was grateful for all the information and warnings I could } lay my hands on before I dealt with any material. The event ,however reported, } serves as a reminder to all of us of the dangers of our materials and the } need to keep up with the information involved in our safety. } This institution has completely reexamined it's glove policy. } } Kate Connolly
Dorothy, You may want to visit the WWW site by Dr. Gwen Childs. It contains the detailed hands on protocols whuch you need and the excellent examples of labelings:ISH and IL. http://cellbio.utmb.edu/childs/childs.htm
Sincerely, Marek Malecki.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
************************ PLEASE POST ************************
Department of Human Resources Trinity College Hartford, Connecticut 06106
Position Announcement
Electron Microscopy Laboratory Manager/ Technician
Trinity College has an immediate opening for an experienced electron microscopist to work in a newly established Electron Microscopy Center. As this facility is designed to serve both the physical and biological sciences, familiarity with both disciplines is highly preferred. Primary responsibilities include assisting faculty and students in the use of the facility for teaching and research, and overseeing the upkeep and use of TEM, STEM and specimen preparation facilities, including the analytical EM EDX and EELS). A B.S. degree with advanced research training and/or extensive experience with TEM are essential; master's degree with requisite experience is preferred.
This position is a full-time, 12-month appointment with full College benefits. Applications will be reviewed upon receipt; search will continue until position is filled. Please respond with a resume, cover letter stating salary expectatons, and the names, telephone numbers and addresses of three professional references to: Prof. Daniel Blackburn, c/o Human Resources, 300 Summit Street, Trinity College, Hartford, CT 06106. (Resumes also may be faxed: (860) 297-5140, or sent via the internet: Sandra.Magee-at-Mail.Trincoll.edu).
Trinity College is an Equal Opportunity/Affirmative Action Employer. Women and minorities are encouraged to apply. Applicants with disabilities should request any needed accommodation in order to participate in the application process.
*****************
Christine Caragianis Broadbridge, Assi. Prof. Trinity College, Department of Engineering 300 Summit Steet Hartford, CT 06106-3100
Forwarded Email please reply direclty to fischg98-at-providence.edu
Below is the result of your feedback form. It was submitted by (fischg98-at-providence.edu) on Tuesday, September 16, 1997 at 11:34:39 ---------------------------------------------------------------------------
Email: fischg98-at-providence.edu Name: Gia Fischetti
School: Providence College
State: RI
Zip: 02918
Question: I am currently doing research on Leishmania enrietti, and I am now tring to prepare specimines for our TEM. I have been unsuccessful at finding a "recipe" for the preparation of this organism. I am reluctant to just try Karnovsky's fixative, and would appreciate any help that you could give me on this matter. Thank you very much. I hope to hear from you soon. Gia Fischetti
If you read the obituary etc., she was clearly quite a scientist. A moral here is check that your gloves are good for what you are using, we have toxicologists/occasional electron microscopists using tributyl tin which is also not nice.
Rick, While the AP is not peer reviewed Science is. Suggest that you check out the August issue. Karen was a collaberator with several people in our lab. Also suggest that you look up "dimethylmercury". One drop (50ul) is equal to about 300X the occupational exposure limit. -- Begin original message --
} } } Without in any way downplaying the tragedy for the people involved, the } dose-response relationship postulated in the story is not very plausible. } Mercury toxicity has been studied extensively for at least half a century. } It is a cumulative poison which does cause the type of neurological effects } reported, but a "single tiny drop", through a glove and quickly washed off, } being lethal seems suspect. Associated Press is not peer reviewed (although } maybe it depends on your definition of "peer"). }
} Rick Mott } rick-at-pgt.com } } (These are personal views having nothing to do with my employer) }
-- End original message --
regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
I'm doing automated particle analysis (SEM/EDX) of suspended particulate material from drinking water reservoirs. Particles are filtered onto nuclepore membranes and carbon coated. Features are located using a thresholded bse image. Diatoms (some species) are so thin that that they don't image very well. Does anybody know of a 'staining' method to increase the atomic number contrast for amorphous silica?
reply to jptmvl-at-mailbox.syr.edu thanx, dave johnson
I would appreciate any information about the software QUANTITEM for quantitative determination of interfacial roughness and composition mappings in HREM images. Thanks in advance.
F. Peiro *******************************+ Francesca Peiro
EME, Enginyeria i Materials Electronics Dpt. Fisica Aplicada i Electronica Universitat de Barcelona Avda. Diagonal 645-647 08028 Barcelona, Spain
The story of the death of Dartmouth College chemistry professor Karen Wetterhahn is unfortunately very true and not greatly exaggerated. She was using very small amounts of dimethylmercury as an NMR standard. The amount reported is "one to a few drops". This is not a large exposure and the exposure occured only once. She was reported to be a very careful chemist. The latex gloves she was wearing did not stop the absorption of the dimethylmercury through her skin. Treatment for mercury poisoning was unsuccessful in saving her life.
Everyone needs to minimize their exposure to toxic materials and microscopists are well aware of this as is noted in this listserver. The composition of gloves worn in the laboratory is very important and each application should be evaluated according to need. It serves no purpose to downplay the potential for tragedy in the lab from toxic materials. Remember also that the greatest potential for problems is the long term exposure to small amounts of materials that the lab person has become accustomed to using - familiarity breeds contempt. Two good articles about Karen Wetterhahn appear in: Chemical and Engineering News, June 16, 1997, p. 11, and Scientific American, September, 1997, p. 20.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The Dartmouth College Biosafety Office in combination with OSHA has produced a comprehensive study /report on the characteristics and abilities of available gloves. On a smaller scale , anyday, I expect a poster for all the labs summarizing the chief points of this work. Anyone wishing information on gloves should contact Michael Blayney at: Michael.Blayney-at-dartmouth.edu
I cannot believe this discussion. No doubt mercury is a nasty cumulative toxin. But blaming a faulty glove and a single drop passing onto the skin is pure nonsense. In the good ol days I cleaned a number of times mercury diffusion pumps and purified the mercury by shaking the stuff with nitric acid and solvents in a separating funnel. No gloves! I have no trace of Minnemata disease, steadier hands than most people and I am "approximately normal". I knew a man who had active mercury poisoning. This was contracted in a large but closed room after a large industrial thermometer fractured on a very hot oven. The man inhaled fumes for several hours. Mercury poisoning most frequently is caused by ingestion and inhalation. I do not advocate bathing in mercury, but that story I take with a drop of something else! Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 400+ Links, MSDS ************************ http://www.proscitech.com.au } } Kate, would you share with us the outcome of your institution's } re-examination of its glove policy? Thanks. /Vachik Hacopian } } } } } As part of the group involved in the postmortem examination of the death of } } Karen Wetterhahn I was grateful for all the information and warnings I could } } lay my hands on before I dealt with any material. The event ,however reported, } } serves as a reminder to all of us of the dangers of our materials and the } } need to keep up with the information involved in our safety. } } This institution has completely reexamined it's glove policy. } } } } Kate Connolly
If I recall correctly, the compound in question was dimethyl mercury, which turned out to pass quickly through her gloves, in contrast to metallic mercury.
} Without in any way downplaying the tragedy for the people involved, the } dose-response relationship postulated in the story is not very plausible. } Mercury toxicity has been studied extensively for at least half a century. } It is a cumulative poison which does cause the type of neurological effects } reported, but a "single tiny drop", through a glove and quickly washed off, } being lethal seems suspect. Associated Press is not peer reviewed (although } maybe it depends on your definition of "peer").
Scott Schwinge Friday Harbor Labs University of Washington
Can we move the Hg discussion to www.Hgtox.com, the gloves discussion to the handwear listserv, and get on to death of more important people (Di)?
Seriously, Prof. Wetterhahn and Di were interesting individuals who have received their eulogies in other more appropriate forums. Whether one drop of diMeHg causes death is debatable, but not here (are any of you microscopists using it?). Gloves get holes and have unsealed wrists (usually). Enough OK? Rob Palmer CEB/UT
If you can still excite the x-ray lines of interest, try lowering the incident beam potential. Woody ______________________________ Reply Separator _________________________________
I'm doing automated particle analysis (SEM/EDX) of suspended particulate material from drinking water reservoirs. Particles are filtered onto nuclepore membranes and carbon coated. Features are located using a thresholded bse image. Diatoms (some species) are so thin that that they don't image very well. Does anybody know of a 'staining' method to increase the atomic number contrast for amorphous silica?
reply to jptmvl-at-mailbox.syr.edu thanx, dave johnson
I get my Osram HBO 100W/2 for a price of $128.00 from
Elkay P.O. Box 1105 La Jolla CA 92038-1105
Larry Kuritzky 619-454-5742
} Does anyone know of a source for the HBO 100W/2 100 watt bulbs } used for fluorescence light microscopy? } } Thanks, } } Dennis Shubitowski } University of Michigan } School of Dentistry } dshubito-at-umich.edu
Dr. Steven Barlow, Associate Director EM Facility/Biology Department 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/EM_Facility
Would anyone know of a used tripod polisher I could buy? I need it to prepare TEM sections of modern and fossil shells (brachiopods) made of calcite.
Thanks.
Nancy Buening
Nancy Buening E-mail: buening-at-geology.ucdavis.edu Department of Geology Phone: (916) 752-0350 University of California Fax: 916 752-0951 Davis, CA 95616
I believe you have missed the point: dimethyl mercury is different from the metallic mercury that you are addressing.
} I cannot believe this discussion. No doubt mercury is a nasty cumulative } toxin. But blaming a faulty glove and a single drop passing onto the skin } is pure } nonsense. } In the good ol days I cleaned a number of times mercury diffusion pumps and } purified the mercury by shaking the stuff with nitric acid and solvents in } a separating funnel. } No gloves! I have no trace of Minnemata disease, steadier hands than most } people } and I am "approximately normal". } I knew a man who had active mercury poisoning. This was contracted in a } large } but closed room after a large industrial thermometer fractured on a very } hot oven. The man inhaled fumes for several hours. } Mercury poisoning most frequently is caused by ingestion and inhalation. I } do not advocate bathing in mercury, but that story I take with a drop of } something else!
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Can we move the Hg discussion to www.Hgtox.com, the gloves discussion to } the handwear listserv, and get on to death of more important people (Di)? } } Seriously, Prof. Wetterhahn and Di were interesting individuals who have } received their eulogies in other more appropriate forums. Whether one drop } of diMeHg causes death is debatable, but not here (are any of you } microscopists using it?). Gloves get holes and have unsealed wrists } (usually). Enough OK? } Rob Palmer } CEB/UT } } }
If you have the option, I would try low kV secondary imaging to do your image analysis, using kV's of 5-10, or even lower. Then later, go to backscatter and increase your accelerating voltage for EDX.
The problem with stains that increase your atomic number contrast, it seems to me, is that they would also greatly affect your EDX results.
Also, you could try doing your EDX first, then sputter-coating your samples for imaging purposes. Of course, then you lose the capability of going back to recheck your x-ray data.
If you must find a particular particle and image and do x-ray on it at the same time, then let me know what you find out from others, because that's a problem I've faced, too. The only thing that comes to mind immediately is image it at low kV, then reconfigure your scope for higher kV backscatter and EDX and hope you don't lose the particle in the process.
Best wishes, Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
Moving the mercury story after the "air " time it has had I can see.
But , I do see a need to discuss gloves on this listserve, we work with too many nasty chemicals. Especially with chemicals such as lowicryl.
Does anyone know of a WWW site for glove permiability information? Perhaps even with microscopy specific chemicals?
I would like to make it a bookmark, or put an anchor to it on our web pages.
Thanks,
Lou Ann
} Can we move the Hg discussion to www.Hgtox.com, the gloves discussion to } the handwear listserv, and get on to death of more important people (Di)? } } Seriously, Prof. Wetterhahn and Di were interesting individuals who have } received their eulogies in other more appropriate forums. Whether one drop } of diMeHg causes death is debatable, but not here (are any of you } microscopists using it?). Gloves get holes and have unsealed wrists } (usually). Enough OK? } Rob Palmer } CEB/UT
*************************** Lou Ann Miller Microscopic Imaging Lab College of Vet. Medicine University of Illinois 2001 S Lincoln Ave Urbana,Illinois 61801 217-244-1566 lamiller-at-ux1.cso.uiuc.edu
Microscopy Home Page: http://www.cvm.uiuc.edu/MicImagLab/MicImagLab.html
Central States Microscopy Society http://www.cvm.uiuc.edu/HomePages/LouAnnMiller/CSMS/csms.html
Personal Home Page: http://www.cvm.uiuc.edu/HomePages/LouAnnMiller/LAM.html
Whether it is proper or not to discuss the death of a scientist in this list, I do not know.
BUT, I do not think a SCIENTIST's death is less valued than those CELEBILITIES' or Di. Without scientists, our world will less progress but without those "celebrities", our world has no impact or even better !!
We are scientists and engineers, we should respect ourselves and be proud of it.
I am not a chemist, but have received quite extensive training on hazardous materials here at work. I have learned that materials not normally absorbed through the skin can be readily absorbed when they are carried by a material that is absorbed. I believe that it may amplify the toxic effects in some cases.
It is dangerous to assume that a material that presents a minor hazard in one state, is not hazardous in another!
David L Johnson wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } I'm doing automated particle analysis (SEM/EDX) of suspended particulate } material from drinking water reservoirs. Particles are filtered onto } nuclepore membranes and carbon coated. Features are located using a } thresholded bse image. Diatoms (some species) are so thin that that they } don't image very well. Does anybody know of a 'staining' method to } increase the atomic number contrast for amorphous silica? } } reply to jptmvl-at-mailbox.syr.edu thanx, dave johnsonDave,
I don't know of an EM approach but there is a very old technique called "Rheinberg Illumination" which works a treat with diatoms. Basically, the technique is called "optical staining" and involves selectively filtering the background versus the scattered light. One of my favorites produces a blue background and yellow diatoms .... a combination which should be pretty effective for automated image analysis systems. We've used them for years on many non-stained specimens, ranging from diatoms to mold in ketsup.
The filters, originally available from KODAK, worked really well with 10x objectives. When they discontinued producing these filters, they were kind enough to give us the originals. We can make a set of about 20 different filters available to you for a modest price. Contact me directly and I can provide you with the details for both how to use these filters and pricing.
Plase let us know how you make out.
Best regards,
Barbara Foster President Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108 PH: (413)746-6931 FX: (413)746-9311 email:mme-at-map.com --------------------------------------------------------------------------------------------------------------------------------- ********** Microscopy/Microscopy Education ********** America’s First National Consortium of Microscopy Experts Specializing in Customized, On-site Training in all areas of Microscopy, Sample Prep, and Image Analysis
What size are the particles you are trying to image?
For the first time I am going to add my two cents worth. In regards to the posting and discussion about dimethylmercury, I would rather read about safety problems and concerns than be subjected to the bickering between list members. This is not the first occasion that some list members have used this public forum to deliver jibes. That the occassion revolves around someone's death makes it all the more objectionable to me. Although Nestor has reminded the entire list of the rules of the list, and the objectionable traffic has abated, I'm seeing it again. Please respect the published purpose and rules of this list. I speak for myself, but perhaps I am not alone in feeling this way.
Sincerely, Maureen Petersen
************************************************************************ Maureen Petersen Department of Plant Pathology 1453 Fifield Hall University of Florida
Dear David, A light gold or gold-palladium sputter coat will increase BSE response and will interfere minimally with EDX identification. Diatoms are pure Si anyway, so small Au and Pd peaks will not interfere. I always use Au-Pd when I image by BSE, unless there are heavy elements already present.
David Johnson wrote: } I'm doing automated particle analysis (SEM/EDX) of suspended particulate } material from drinking water reservoirs. Particles are filtered onto } nuclepore membranes and carbon coated. Features are located using a } thresholded bse image. Diatoms (some species) are so thin that that they } don't image very well. Does anybody know of a 'staining' method to } increase the atomic number contrast for amorphous silica? } } reply to jptmvl-at-mailbox.syr.edu thanx, dave johnson Regards, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
I keep Paraplast Plus melted in a bath more or less continuously, so it is always ready to use, even though it may be one or two months between uses. Lately my students have been having some sectioning problems (with freshly sharpened blades) and before I suggest that maybe they have done something wrong during embedding, is it possible that the wax has "gone bad?"
Gary Radice, Associate Professor gradice-at-richmond.edu Department of Biology 804-289-8107 (voice) University of Richmond VA 23173 804-289-8233 (FAX)
I always immerse the grids for IEM-colloidial gold expt's. That is if I am using only 1 primary antibody. If using 2 primary antibody, then I float the grids, one side per antibody/gold. As a side note, I also immerse the grids when staining with UA and Pb.
Best of Luck Ed Calomeni Medical College of Ohio Dept Pathology Toledo, OH 43614 emlab-at-opus.mco.ecu
Colleagues , I can't help this guy can any of you? Reply back to him not the listserver...
Nestor Your Friendly Neighborhood SysOp
Below is the result of your feedback form. It was submitted by (dmatthew-at-providence.edu) on Sunday, September 14, 1997 at 20:46:01 ---------------------------------------------------------------------------
Question: Hello. I am an undergrad student at PC just beginning a class using the EM. For a project I plan on examaning the amebocytes of limulus polyphemus (horshoe crab). I am intersted in any information on fixation techniques of the cells for the TEM. I had planned on centrifuging blood and then processing the pellet. I've begun a literature search for specific techniques, but getting my hands on the more obscure journals can be difficult. Any help would be appreciated. Thanx so much.
We got our target from Goodfellow Metals in Cambridge UK Tel: +44-1223-568068 Fax: +44-1223-420639. They have high purity chromium foils of various thicknesses and will laser cut to what ever size is needed. We ordered a 1mm thick target for the Denton Magnetron Sputter Coater we have on our Oxford Instruments CM1500 cryopreparation unit. It works very well. Get Goodfellows to send you their very informative catalogue.
Patrick Echlin Director, Multi-Image Centre University of Cambridge
On Wed, 10 Sep 1997, T. Graham wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } I need to get ahold of a chromium sputtering target. Does anyone know } which company I may purchase this from? } } Tom Graham } } }
Does anyone know of the Histoclear and Histomount clearing and mounting media? I'd like to know the name of the manufacturer and where they are located? Thanks,
Imre Kovacs M.D. kimre-at-comser.szote.u-szeged.hu
Alzheimer's Disease Research Center Albert Szent-Gyorgyi Medical University H-6720 Szeged, Somogyi u. 4. Hungary
Dear colleagues, I have formvar powder and choloform at my disposal to prepare holey carbon films. Could you give me the method to use or the references of some literature where I can find it? Thank you for your help! Nathalie Bozzolo _________________________________________________
Nathalie Bozzolo Laboratoire d'Analyse des Materiaux Centre de Recherche Public - Centre Universitaire 162a, avenue de la Faiencerie L-1511 Luxembourg tel : (352)46 66 44 402 fax : (352)46 66 44 400 _________________________________________________
Many thanks to Phil Dahlstrom for sorting out the "mercury" confusion. It reminds me of the time when I was an inexperienced young scientist at the Paint Research Association (and dinosaurs walked the earth!). I had thought of using tetraethyl lead as an (electron? X-ray?) dense material, and was forcefully warned off by one of the senior staff there. Simply because it was found in petrol did not mean that it was OK to use: in fact, at the "lead" factory there would be paraffin (kerosene) showers, and on the slightest contact the victim would be shoved under, clothes and all - speed is of the essence.
More recently, I was looking for ways of putting heavy metal into specimens either for TEM density or for backscattering SEM, and I discovered recent work in the literature, suggesting triphenyl bismuth as a relatively low toxicity material - bismuth compounds are apparently remarkably innocuous compared to its neighbours in the periodic table. I didn't get so far with that, as we found another way of imaging our specimens - water trees in electric cables - but if anybody is interested, here are a couple of references they could follow up:
(1) TI: THERMOMECHANICAL INVESTIGATION OF POLY(METHYLMETHACRYLATE) CONTAINING AN ORGANOBISMUTH RADIOPACIFYING ADDITIVE AU: RAWLS_HR, GRANIER_RJ, SMID_J, CABASSO_I JN: JOURNAL OF BIOMEDICAL MATERIALS RESEARCH, 1996, Vol.31, No.3, pp.339-343
(2) TI: Radiopaque copolymers of styryldiphenylbismuth vinylbenzylphosphonate and methyl methacrylate AU: Tamber_H, Smid_J, Cabasso_I JN: CHEMISTRY OF MATERIALS, 1997, Vol.9, No.6, pp.1335-1341
+------------------------------------------------------------------------+ | 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 | +------------------------------------------------------------------------+
} -----Original Message----- } From: Vladimir Dusevich [SMTP:dusevich-at-astro.ocis.temple.edu] } Sent: Friday, September 19, 1997 2:56 PM } To: Robert J. Palmer Jr. } Cc: Microscopy-at-Sparc5.Microscopy.Com } Subject: Re: Hg, gloves, death } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } ---------------------------------------------------------------------- } -. } } 100% agree!!! } } On Fri, 19 Sep 1997, Robert J. Palmer Jr. wrote: } } } } ---------------------------------------------------------------------- } -- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } } ---------------------------------------------------------------------- } -. } } } } Can we move the Hg discussion to www.Hgtox.com, the gloves } discussion to } } the handwear listserv, and get on to death of more important people } (Di)? } } } } Seriously, Prof. Wetterhahn and Di were interesting individuals who } have } } received their eulogies in other more appropriate forums. Whether } one drop } } of diMeHg causes death is debatable, but not here (are any of you } } microscopists using it?). Gloves get holes and have unsealed wrists } } (usually). Enough OK? } } Rob Palmer } } CEB/UT } } } } } }
INVITATION TO SUBMIT PROPOSALS FOR 1) PROGRAM PARTICIPATION and 2) 1998 FACULTY FELLOWSHIPS
The Shared Research Equipment (SHaRE) User Facility and Program at the Oak Ridge National Laboratory (ORNL) provides access to a variety of advanced instrumentation for collaborative materials science research. Through the SHaRE User Program, materials scientists from universities, industries, or other government laboratories may access the SHaRE Facility as well as other instrumentation within ORNL's Metals and Ceramics (M&C) Division. Facility instrumentation includes a variety of electron microscopes, atom probe field-ion microscopes, and mechanical properties microprobes.
SHaRE Program Participation Proposals are being solicited at this time for facility use during fiscal year 1998 (October 1, 1997 - September 30, 1998). The program is intended to support collaborations between M&C staff members and researchers external to ORNL. Therefore, proposals must identify at least one staff member and one non-ORNL participant who will act as a co-principal investigator and share responsibility for the project. Proposals will be reviewed by a committee and evaluated with regard to scientific excellence, relevance to the interests of the U.S. Department of Energy, Division of Materials Sciences, and the likelihood of project success. Additionally, principal investigators and graduate students from U.S. accredited universities are eligible to receive funds to defray certain program-related travel and subsistence expenses.
The SHaRE Facility instrumentation and the related User Program are described in detail at http://www.ornl.gov/share. In the past, only letter proposals were submitted for program participation. However, application for program participation and travel support is now made by using a downloaded form located at http://www.ornl.gov/share/pdf/proposal98.pdf. The form will be mailed to potential applicants upon request.
1998 Faculty Fellowships Faculty fellowships provide outstanding university faculty extended access to the SHaRE User Facility at ORNL. It is anticipated that at least one junior and one senior university faculty member will be appointed as fellows. Information regarding the fellowships, including eligibility requirements, length of appointment, stipends, and application guidelines may be found at http://www.ornl.gov/share. The guidelines will be mailed to potential applicants upon request.
SHaRE will accept and review proposals for projects and fellowships at any time during the fiscal year, but allocates the majority of funds during the month of October. Proposals for review during the October meeting should be received at the address below by October 10, 1997. Proposals will be reviewed, and travel awards announced, in mid-October. Fellowship applications received after the October date will be reviewed during February 1998.
Proposals submitted (5 copies) should be sent to:
SHaRE Proposals Education and Training Division, MS-36 Oak Ridge Institute for Science and Education P.O. Box 117 Oak Ridge, Tennessee 37831-0117
SHaRE is jointly administered for the U.S. Department of Energy by ORNL and the Oak Ridge Institute for Science and Education (ORISE). For additional information or clarification on proposal submissions, please contact me directly.
Regards,
Neal
Dr. Neal D. Evans voice: (423) 576-4427 Shared Research Equipment Program facsimile: (423) 574-0641 Oak Ridge National Laboratory email: evansnd-at-ornl.gov Building 5500, MS 6376 Oak Ridge, TN 37831-6376
Greetings! I am looking for a site where I can find microscopic pictures of various kinds of protists for my fifth grade science students. I have drawings, but we do not have access to slides of the real thing. Perhaps someone out there can direct me to a useful website that will interst my students. Thanks! Reply to: jpolak-at-esu6.esu6.k12.ne.us Julie Polak Exeter Public Schools Exeter, Nebraska
} I have to prepare some yeast for SEM. The person who requested this } work wants some pretty pictures of his yeast, Schizosaccharomyces pombe, } for use in seminars. I have read up on some techniques to use but have } two main queries: } 1. To collect the yeast onto filter paper ( a method used in } several publications), do I just drop a suspension of the yeast onto the } filter paper? What sort of filter paper do I use? Is there a "better" } way of collecting these cells such as settling them onto poly-l-lysine } coverslips?
We use regular microscope slides, cleaned well (detergent, acid or alcohol): coat with l mg/ml aqueous solution of poly-L-lysine for several minutes, rinse in distilled wate.
Take a (preferably) aqueous suspension of the yeast and place onto the microscope slide and allow the cells to settle for about one hour at RT. Carefully tip the slide and allow the unattached cells to flow off. Now, gently dropper some fixative (2% glut/4% formald in buffer of choice) onto the slide to completely cover the smear. Allow to set undisturbed overnight at RT in a humid chamber (petri dish with filter paper or Tupperware container with moist paper towels). Transfer the slide into a rinse solution (Coplin jar of distilled water or petri of distilled water). With some yeasts, the aldehyde fix is adequate to preserve the integrity of the cells (others may collapse without osmium post-fixation). The slides are then slowly dehydrated in ethanol (20-40-60-80-100-100%) for 10 min each. Critical point dry using liquid carbon dioxide as transitional solvent. You will need to break the slide into 1 inch squares to fit into the CPD device. Do this by scoring the slide with a diamond marker pen and pressing down gently onto an applicator stick. Mark an "X" the back side with the diamond marker to help identify the good side later. Freeze drying from the ethanol should also work well.
IF osmium is needed place slide/specimen into 2% aqueous osmium tetroxide overnight - take care to avoid evaporation by either immersion of the slide into a shallow container of osmium or by vapor fixation of the slide/specimen. Vapor fixation (CAUTION WITH OS FUMES - DO IN PROPERLY OPERATING FUME HOOD) may be accomplished by placing the wet slide/specimen into a plastic petri dish and then placing a small volume (4-5 ml) of 2% osmium solution nearby in the dish. Rinse in distilled water and dehydrate and CPD as described above.
Caveats: do not overload the slides with culture since you do not want a heaped up mess but isolated cells. A suspension that is quite turbid should work well - not a paste or milky/opaque solution.
Contact me if you have any other questions. I have done a lot of imaging of yeasties by TEM and SEM.
Cheers,
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing 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 would like to ask you for your advise on cryo-sectioning of plant material in possibly low temperature. We tried to section the frozen piece of leaf using Cryocut E Reichert, playing with different embedding media but the results were not satisfactory. We found difficulties in cutting thick sections (5-30 microns) even of very young leaf. Morphological structure was not well preserved.
I really would appreciate your advise, suggestions, tips and technical tricks in this matter. What equipment do you use and what can you recommend?
Thank you in advance for all your time and assistance
With best regards
Jolanta Mesjasz-Przybylowicz ************************************************************************ Dr Jolanta Mesjasz-Przybylowicz National Accelerator Centre P.O. Box 72 Faure 7131 South Africa tel: 27-21-8433820 fax: 27-21-8433543 Internet: MESJASZ-at-nac.ac.za ************************************************************************
I have used both with excellent results, for 10 micron and 100-200 micron sections. Histomount *really* likes to shrink, so it needs to be watched while drying. Also, it makes zillions of air bubbles if you try to dry it with heating like is done with Permount. Dry finished slides at room temp. Excess Histomount cleans up with Histoclear and a razor blade.
Both are made/sold by National Diagnostics in New Jersey, US, but I don't have the contact information. (This is a year old--I assume the information hasn't changed.)
Fisher sells a similar (identical?) product to Histoclear, but I don't remember if they have a Histomount analog.
Phil
} Does anyone know of the Histoclear and Histomount clearing and } mounting media? I'd like to know the name of the manufacturer and } where they are located? } Thanks, } } } Imre Kovacs M.D. } kimre-at-comser.szote.u-szeged.hu } } Alzheimer's Disease } Research Center } Albert Szent-Gyorgyi } Medical University } H-6720 Szeged, } Somogyi u. 4. } Hungary
1. Add 4 drops of solution b to solution a, and shake vigorously for 30 seconds. 2. Pour on clean glass slides and dry. (ie: dip the slides into a staining dish.) 3. Breath on slide several times. 4. Float layer off the slides. 5. On the floating layers, gently place grids, dull side down. 6. Pick up layer with parafilm, letting the film stick to the parafilm. 7. Let dry for 30 minutes. 8. Put in petri dish with filter paper saturated with 50% methanol for 20 minutes. 9. Take each grid off mesh and dip in 50% methanol and place on filter paper to dry. 10. Coat with carbon in a vacuum evaporator. 11. Immerse each grid in chloroform briefly.
} ---------- } From: bozzolo-at-crpcu.lu[SMTP:bozzolo-at-crpcu.lu] } Sent: 22 September, 1997 06:11 } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: TEM - holey carbon films } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi The company is:National Diagnostics 404-699-2121
bob
On Mon, 22 Sep 1997, IMRE KOVACS M.D. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear Microscopists, } } Does anyone know of the Histoclear and Histomount clearing and } mounting media? I'd like to know the name of the manufacturer and } where they are located? } Thanks, } } } Imre Kovacs M.D. } kimre-at-comser.szote.u-szeged.hu } } Alzheimer's Disease } Research Center } Albert Szent-Gyorgyi } Medical University } H-6720 Szeged, } Somogyi u. 4. } Hungary } }
} Rick, } While the AP is not peer reviewed Science is. Suggest that you check out } the August issue. Karen } was a collaberator with several people in our lab. Also suggest that } you look up } "dimethylmercury". One drop (50ul) is equal to about 300X the } occupational exposure limit. } -- Begin original message -- } } Without in any way downplaying the tragedy for the people involved, the } } dose-response relationship postulated in the story is not very plausible. } } Mercury toxicity has been studied extensively for at least half a century. } } It is a cumulative poison which does cause the type of neurological effects } } reported, but a "single tiny drop", through a glove and quickly washed off, } } being lethal seems suspect. Associated Press is not peer reviewed (although } } maybe it depends on your definition of "peer"). } } } } Rick Mott } } rick-at-pgt.com } } } } (These are personal views having nothing to do with my employer) } } } } -- End original message -- } } } regards, } Bob } Robert Schoonhoven
Of course, one of the problems is that the original newspaper article that started this thread refered to 'mercury'. Which as many have pointed out, as elemental metallic mercury while dangerous is essentially a long term, cumulative poison and there is no possibility that single drop on even unprotected skin will cause the slightest harm.
On the other hand, dimethylmercury is an entirely different kettle of fish (apologies for colloquialism). This is a classic example on the confusion that arises when the scientifically ignorant start talking about things they don't understand.
It reminds me of the debate in the British parliament a number of years ago on the merits of fluoridation of water - the principle case was made by a scientifically ignorant MP who based his argument against fluoridation on a dictionary definition of the chemical and biological properties of fluorine. One wonders if he had ever looked up the same information for chlorine and took a similar stand on his use of salt on his food.
While there are many arguments in science, particularly in areas relating to ethical issues, which may be open to the general public, there are equally many that aren't. It is simply a case that if you haven't studied and learnt the basic facts, you are ignorant and aren't qualified to comment. And notions of democracy, and free speech don't change that.
The Division of Nephrology in the University of Florida College of Medicine is seeking a full-time electron microscopy technician. This individual will be one of two full-time electron microscopy technicians staffing the Division's Electron Microscopy Facility, which serves as a research facility for the members of the Division of Nephrology and their collaborators. The research conducted primarily strives to determine correlations between renal ultrastructure and function using transmission and scanning electron microscopy, morphometric analysis, and immunogold and immunoperoxidase cytochemistry. Equipment housed within the facility include 2 Zeiss EM-10A transmission electron microscopes, a Topcon DS130C scanning electron microscope, 2 LKB Nova ultramicrotomes, a Leica Automatic Freeze Substitution unit, and all related support equipment. The candidate's duties will include tissue processing, ultramicrotomy, immunocytochemistry, routine maintenance of the electron microscopes, viewing samples, photographic processing, and related support functions. The candidate may be hired as an Electron Microscopy Technician of Senior Electron Microscopy Technician depending on experience.
Reply to Dr. Jill Verlander verlandr-at-medicine.ufl.edu ******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab PO Box 118525 Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
Our Ohio company is seeking engineering / scientist support for microelectromechanical systems research & development. Work will involve new materials and surface treatments to control the friction and wear of MEMS devices; Friction & wear measurement of MEMS systems / materials.
It is important that candidates have capabilities in cross-section TEM, analytical TEM of tribological materials; especially wear tracks; creation of unique microstructures; and understand friction & wear on a fundamental level.
Contact Ronald Decker - deckerrc-at-utcdayton.com
Ronald C Decker Program Manager Universal Technology Corporation 1321 Research Park Drive, Suite 100 Dayton OH 45432-2817 Voice (937) 426-8530, Fax (937) 426-7753
Dear All: I inherited a used Jeol 100ZX. It is in excellent working condition and about 20 years old, located in the Los Angeles area. If you are in need of a TEM or want it for spare parts please make me an offer. Peter Jordan, EMSI 909 694-1939
This information is correct as of March 95 when we last ordered. A European source of Hystomount (and I think also Hystoclear) is:
Hughes and Hughes Ltd. Unit 1F, Lowmoor Industrial Estate, Tonedale, Wellinton, Somerset TA21 0AZ England Phone +44 823 660222 FAX +44 823 660186
As regards to Hystoclear, there are other companies which sell a limonene-based xylene substitute. As an example, Fisher calls its product Hemo-De. They are different than xylene in more than just the toxicity. Thus I would ask for a small sample from your supplier to try before you buy. In my experience these products are normally sold in large quantities. (Fisher lists it smallest size as 1 gallon).
Good luck, Azriel Gorski
On Mon, 22 Sep 1997, IMRE KOVACS M.D. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear Microscopists, } } Does anyone know of the Histoclear and Histomount clearing and } mounting media? I'd like to know the name of the manufacturer and } where they are located? } Thanks, } } } Imre Kovacs M.D. } kimre-at-comser.szote.u-szeged.hu } } Alzheimer's Disease } Research Center } Albert Szent-Gyorgyi } Medical University } H-6720 Szeged, } Somogyi u. 4. } Hungary }
We need a Zeiss Axioplan microscope for transmitted light and fluorescence work. This would be the original Axioplan, not the new Axioplan 2 model. If you own this microscope, and are interested in selling or trading your instrument, please send me details of the configuration you have. Part numbers would be helpful.
Patrick
-------------------------------------------------------------- Dr. Patrick L. Huddie (301) 725-2775 Fax (301) 725-2941 Microcosm, Inc., 9140 Guilford Road, Suite O, Columbia, MD 21046 e-mail phuddie-at-microcosm.com URL http://www.microcosm.com
I want to attach a video system to my light microscope. I just bought the camera (Sony ccd 370) and the corresponding adaptor and lens, although now I'm having trouble with the board. I bought a miro dc30 board but I only get a small image (I want the image on the entire screen), and, besides, I'm not having good printings. I'm working with and HP 133 Mhz, 48 MB ram, and my printer is an HP 870. I guess that this are good working conditions, thus, I believe that the board is not correct. Can anyone help me? Thank you in advance!
I would check out the adapter and lens system before the board.
We have a Pixera camera that we have tried to replace our old RS-170 camera with. The lens/adapter for the RS-170 camera screws right in to the forn of our Pixera, however, the field of view is only about one third the size it was with our RS-170 camera. I came to find out that that should not be a surprise. The chip on the Pixera is only 1/3" across while it is 1" across on the RS-170. We have yet to get the right adapter, but have looked at items from Optem, Diagnostic Instruments, and Edmund Scientific. We should be able to get much closer.
There are others out there that have gone through the same exercise. Maybe they will speak up too.
At 08:21 AM 9/23/97 -0300, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear microscopists: We have very good deal of barely used, looks like new NORAN confocal and RMC ultramicrotome and their accesories for sale. Please contact Dorothy at 617-432-2970. Thanks.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I want to attach a video system to my light microscope. I just bought } the camera (Sony ccd 370) and the corresponding adaptor and lens, } although now I'm having trouble with the board. I bought a miro dc30 } board but I only get a small image (I want the image on the entire } screen),
Not necessarily. I am not familiar with your camera or board but the problem could be that the relay lens for the camera has too large a field of view. Have you tried connecting the camera directly to a monitor? Is the image acceptable on that?
Thanks to all who responded to my question about my Denton not sucking. The problem turned out to be a broken rough pump bellows. Ida (the parts lady) at Denton was great! Even though there was a long backorder for the part, she searched all over Denton and found a bellows for me. She did this out of the kindness of her heart. Thanks to her we were able to repair my machine lickety-split. I just thought I'd let everybody know how much I appreciate them, the BBer's for advice and the Denton folks for being so helpful. My Denton now sucks like you wouldn't believe. It hasn't worked this well in a long time. It can now pump down to 2 X 10-5 without any LN2 added to the trap. I guess the moral of the story is...It always pays to ask those who know.
Happily carbon coating in Berkeley,
Paula = )
p.s. I have no financial interest in Denton, etc, etc, etc.
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
I am not familiar with the Sony system, but if it is NTSC video, then you will be limited to about 500 lines of vertical resolution. Horizontal resolution will depend on the Sony and the capture card. It is probably about 400 lines... This will equate to 500x~400 pixel image. If your computer screen is set to - say 1024x768 pixel display, the unmodified image would fill about 1/2 the CRT. There may not be a convenient way to increase the Sony resolution, but you could fill the screen by reducing the CRT resolution to 640x480. Better yet, (although "hollow" magnification) would be to increase the image pixel array size (software manipulation) after capture to fill the screen. Increasing the pixel array for the same size hard copy may help produce better halftone printer output also.
For work like this, I would prefer higher resolution, digital still cameras. The obvious drawback is lack of "real-time" output.
Woody White, Electron Microscopist SEM/EDS/WDS
Work: Mcdermott Technology, Inc. woody.n.white-at-mcdermott.com http://www.mtiresearch.com/
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I want to attach a video system to my light microscope. I just bought the camera (Sony ccd 370) and the corresponding adaptor and lens, although now I'm having trouble with the board. I bought a miro dc30 board but I only get a small image (I want the image on the entire screen), and, besides, I'm not having good printings. I'm working with and HP 133 Mhz, 48 MB ram, and my printer is an HP 870. I guess that this are good working conditions, thus, I believe that the board is not correct. Can anyone help me? Thank you in advance!
Our lab is in the transition of doing immunogold tagging in HRLM-TEM.But we continue to do immunofluorescent techniques received in Michels transport media. Is there anyone out there still doing immunofluorescent techniques (direct or indirect immunofluorescent techniques) on renal, skin, muscle or nerve biopsies? If yes, is the biopsy snap-frozen in liquid nitrogen as soon after removal as possible (precooling a metal chuck for about a minute. Placing a small volume of saline or water on top of the colled chuck, immediately placing the renal or skin bx into the water or saline drop. As the water and the bx begin to freeze, the chuck is turned down to eliminate excess water or saline. The chuck is placed in liquid nitrogen for approx one minute to snap-freeze the bx or tissue. When nitrogen stops bubling bx is adequately frozen and bx is ready to be stored at -70oC or to be sectioned). If not possible to do technique is the tissue being storred in Michels' transport media also sold commercially as Zeus. Are the renal biopsies being tagged with IgG, IgM, IgA, C3, C1q, Kappa & Lambda? Skin, muscle and nerve biopsies tagged with IgG, IgM, IgA, C3 and Fibrinogen? Please respond. Thanx Teresa
Our lab is in the transition of doing immunogold tagging in HRLM-TEM.But we continue to do immunofluorescent techniques received in Michels transport media. Is there anyone out there still doing immunofluorescent techniques (direct or indirect immunofluorescent techniques) on renal, skin, muscle or nerve biopsies? If yes, is the biopsy snap-frozen in liquid nitrogen as soon after removal as possible (precooling a metal chuck for about a minute. Placing a small volume of saline or water on top of the colled chuck, immediately placing the renal or skin bx into the water or saline drop. As the water and the bx begin to freeze, the chuck is turned down to eliminate excess water or saline. The chuck is placed in liquid nitrogen for approx one minute to snap-freeze the bx or tissue. When nitrogen stops bubling bx is adequately frozen and bx is ready to be stored at -70oC or to be sectioned). If not possible to do technique is the tissue being storred in Michels' transport media also sold commercially as Zeus. Are the renal biopsies being tagged with IgG, IgM, IgA, C3, C1q, Kappa & Lambda? Skin, muscle and nerve biopsies tagged with IgG, IgM, IgA, C3 and Fibrinogen? Please respond. Thanx Teresa
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } I would check out the adapter and lens system before the board.
Agreed.
} We have a Pixera camera that we have tried to replace our old RS-170 camera } with. The lens/adapter for the RS-170 camera screws right in to the forn of } our Pixera, however, the field of view is only about one third the size it } was with our RS-170 camera. I came to find out that that should not be a } surprise. The chip on the Pixera is only 1/3" across while it is 1" across } on the RS-170. We have yet to get the right adapter, but have looked at } items from Optem, Diagnostic Instruments, and Edmund Scientific. We should } be able to get much closer.
I am not sure which particular kind of microscopy will suit my needs, and would very much appreciate it if somebody could help me in this regard.
My application is as follows: I need to look at molecular-level changes occurring at a gas-liquid interface with respect to some parameters. The liquid will have, among other components, proteins in it. I want to look at the larger molecules which are affected by an interface. For this, I plan to take a liquid volume and disperse bubbles in them. Then I plan to freeze a part of the liquid rapidly and carry out imaging at various sections. I don't know what kind of technique or microscopy is the best for this. I would appreciate comments and suggestions on this. Also, if somebody knows of a paper or book that has this kind work in it, then that would give me some leads. I have searched some of the indexes with keywords like surface, bubble, modification, microscopy, protein etc., but could not find what I wanted. Am I missing something here? We have some generic metrology equipment like Surface profilers (contact and Optical) , SEM, AFM, Light Microscopes (not near-field). I would also be interested in knowing if any of this can be adapted for my needs. I have NOT done any of the following: biological tissues examining/sectioning, staining, fixing, cryomicroscopy, AFM etc. My experience is with microfabricated structures and examination under the SEM, light microscope and surface profilers. However, I am very willing to learn a new field, and can work towards obtaining a new piece of equipment, or try to have some arrangement with interested commercial/non-commercial parties.
Thanks very much.
Ashok Institute for Micromanufacturing Louisiana Tech University E-mail: krishnan-at-engr.latech.edu ph: (318) 251-8110, fax: (318) 257 5104
"Smaller, lighter, more functional and less expensive consumer products, industrial machines, instruments...possibilities limited only by man's imagination"
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Ms. Polak: Please look at the website list that is section V of the Project MICRO bibliography (address below). And if you're really into protozoa, I recommend the recent book by Anderson & Druger listed in section IIB.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
The Analytical Microscopy Core facility at Hospital for Special Surgery, New York, NY, is searching for a full time electron microscopy technician. The Core provides TEM and SEM service to about 50 clinical and basic scientists, whose major interests are in orthopedics and connective tissue diseases. About 2 days per week would be used to provide technical assistance to a senior researcher, which would involve TEM, SEM, immuno and histochemistry, and development of image analysis techniques. This individual should be familar with basic TEM and SEM techniques but will be trained to work with connective tissues and calcified cartilage and bone. The job can be classified as EM Technician or Senior Technician, depending on qualifications. Interested persons please respond to Steve Doty at address below:
Stephen B. Doty, PhD. Phone: (212) 606-1417 Director, Analytical Microscopy Core Fax: (212) 717-1192 Hospital for Special Surgery email: dotys-at-hss.edu 535 E. 70th Street, NY, NY 10021
Dear All: I made two mistakes on my posting yesterday, first the Jeol is a 100C and not a 100ZX and my phone number is 909 694-1839 and not 1939. The thing I got right was that it is 20 years old and it is still for sale. Sorry, I'll never write an e-mail letter at midnight. Peter Jordan, EMSI
Dear All: A few months ago there was a posting about a Zeiss 9 TEM given away for free. If this is still available or if you know who did the posting please let me know. If I remember right it was in the San Diego area. Thank you. Peter Jordan
Greetings again! I want to thank all of you terrific people out there who have so graciously sent me information about protist pictures!!!! I took my 5th graders to several of the sites and they said the protists were "AWESOME!!!" (I concurred!) We especially liked the "zoo." Several of you sent personal messages encouraging my students and me, and also sent links and other interesting information. Thanks again for all your help! I think I've even made a couple of new net-friends! Many thanks, Julie Polak
At 10:32 AM 9/24/97 +0200, you wrote: } I have a problem with an import the file with an extension *.sp ( spectrum ) } and put it to the document in word 6 or amipro } } Malgorzata Warmuzek
As a rule, you must have a program that supports OLE (object linking and embedding) for the type of file that you have at hand before you can import that file into another program. I suspect that you have no such program for your SP files.
You will probably have to export the file to some form that can be read by your spreadsheet or graphing program, prepare your graph there, and then copy that graph into your word processor. I haven't worked much with Microsoft's MSGRAPH that comes with Word, but it might do what you need in a rudimentary sort of way without invoking a spreadsheet program. ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering, or 270 Metals Development Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-3091
We had so much fun yesterday at the Microbe Zoo and at Kunkel's Gallery, that I want to share our four favorite sites with the rest of the world (as it were). I think these would be helpful to other upper elementary/middle school teachers and students.
I use Thumbs Plus (approx. $60 US if I remember correctly) Cerious Software, Inc. 1515 Mockingbird Lande Suite 209 Charlotte, NC 28209 http://www.cerious.com
I have no financial interest in this company other than doing my part as a very satisfied customer.
} ------------------------------------------------ } Opinions or statements expressed herein, rational or otherwise, do not } necessarily reflect those of my employer. } } Harold J. Crossman } OSRAM SYLVANIA INC. } Lighting Research Center } 71 Cherry Hill Dr. } Beverly, MA 01915 } Phone: (508) 750-1717 } E-mail: crossman-at-osi.sylvania.com } } Our web sites: www.sylvania.com } www.siemens.com } -- } } "Crossman, Harold" {crossman-at-osi.SYLVANIA.com} } } }
We are in the process of resurrecting Cambridge 250 Mk3 SEM and we are looking for the source of spare parts. Any feedback on the subject will be greatly appreciated.
As far a s cryosections of leaves are concerned. It ain't easy because of all the internal air spaces. Cryofracturing is no problem and with a kittle care you can cryoplane the leaf ti get a nice smooth surface. I think I know what you want to do Jolanta (are you still doing ion beam microscopy ?) May be freeze substitution is the best approach. If all else fails read my book
Best wishes
Patrick Echlin University of Cambridge
On Mon, 22 Sep 1997, Jolanta Mesjasz-Przybylowicz wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } } Dear Microscopists, } } } I would like to ask you for your advise on cryo-sectioning of plant } material in possibly low temperature. We tried to section the frozen } piece of leaf using Cryocut E Reichert, playing with different embedding } media but the results were not satisfactory. We found difficulties in } cutting thick sections (5-30 microns) even of very young leaf. } Morphological structure was not well preserved. } } I really would appreciate your advise, suggestions, tips and } technical tricks in this matter. } What equipment do you use and what can you recommend? } } Thank you in advance for all your time and assistance } } With best regards } } Jolanta Mesjasz-Przybylowicz } ************************************************************************ } Dr Jolanta Mesjasz-Przybylowicz } National Accelerator Centre } P.O. Box 72 } Faure 7131 } South Africa } tel: 27-21-8433820 } fax: 27-21-8433543 } Internet: MESJASZ-at-nac.ac.za } ************************************************************************ }
Thumbs plus by Cerius Software works very well for us as an image filing tool. It provides a thumbnail of all images in a directory.
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-bgsm.edu * **************************************************************
On Wed, 24 Sep 1997 mark_munro-at-bio-rad.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } } Dear all, } does anyone know of any shareware/inexpensive image database and } archiving software. } } Thanks in advance, } } Mark Munro } } }
mark_munro-at-bio-rad.com wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Dear all, } does anyone know of any shareware/inexpensive image database and } archiving software. } } Thanks in advance, } } Mark Munro
Mark,
Company Cerious Software Inc. http://cerious.catalogue.com/index.html has a great software ThumbPlus 3.0 for 60 US$.
Henrik Kaker SEM-EDS Laboratory Metal Ravne d.o.o. Slovenia
We are looking for any information we can find on the microscopy/identification of particles from automobile tires. Seems we have a case where we need to try to match particles from a pair of tennis shoes to a particular set of automobile tires(best case), or at least identify particles as possibly coming from automobile tires (more likely). (I'll let your imagination fill in the details on this one!)
If anybody has done anything like this, please let us know. In the meantime, we'll be hitting the indexes/abstracts and databases.
Never a dull moment.... Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
ThumbsPlus is very good. I've used a single site license. In my new position, I've just gotten 5 concurrent site licenses for myself and my team. I don't know how good their Mac version is. I believe that it is still in beta testing. It is shareware and Cerious software has a website where you can download the shareware version. Once you are registered, there are some things that are available, but the unregistered version is very good. I tried it out that way after it was suggested previously on the Microscopy Listserver.
-Scott Walck
"The opinions expressed are those of S.D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
We do direct immunofluorescence on renal and skin biopsies. For the most part, the specimens are received in Michels transport media (which we make ourselves). Upon arrival, the specimens are washed in 3 changes of transport buffer to flush out the excess salts of the michels media (3 washes, 10 minutes each, room temp). The specimen is then lightly blotted, oriented in a cryomold, covered in OCT, and frozen. To freeze, we cool iso-pentane (2-methylbutane) in liquid nitrogen, and lower the cryomold into the cooled iso-pentane for a few seconds. We then attach a chuck in the cryostat, and cut. When the specimen is received fresh, we skip the washing part, and proceed as above. On the renal biopsies, we run: IgG, IgA, IgM, C3, C1q, Fibrinogen, Kappa, Lambda, and Albumin. On the skin biopsies, we run: IgG, IgA, IgM, C3, C1q, Fibrinogen, and Albumin.
Hope this answers your question.
Phyllis Davie Immunocytochemistry Laboratory University of Washington Medical Center pdavie-at-u.washington.edu
On Tue, 23 Sep 1997, Flores, Teresa wrote:
} Our lab is in the transition of doing immunogold tagging in HRLM-TEM.But we } continue to do immunofluorescent techniques received in Michels transport } media. Is there anyone out there still doing immunofluorescent techniques } (direct or indirect immunofluorescent techniques) on renal, skin, muscle or } nerve biopsies? If yes, is the biopsy snap-frozen in liquid nitrogen as } soon after removal as possible (precooling a metal chuck for about a } minute. Placing a small volume of saline or water on top of the colled } chuck, immediately placing the renal or skin bx into the water or saline } drop. As the water and the bx begin to freeze, the chuck is turned down to } eliminate excess water or saline. The chuck is placed in liquid nitrogen } for approx one minute to snap-freeze the bx or tissue. When nitrogen stops } bubling bx is adequately frozen and bx is ready to be stored at -70oC or to } be sectioned). } If not possible to do technique is the tissue being storred in Michels' } transport media also sold commercially as Zeus. } Are the renal biopsies being tagged with IgG, IgM, IgA, C3, C1q, Kappa & Lambda? } Skin, muscle and nerve biopsies tagged with IgG, IgM, IgA, C3 and Fibrinogen? } Please respond. Thanx Teresa } } } }
You can capture the spectrum with a software, such as Paint Shop Pro we are using, and save it as TIFF or Bitmap file. If you have the software, I can show you the details.
With the best wishes, Charlie Kong kong-at-t-rex.materials.unsw.edu.au
On Wed, 24 Sep 1997, Malgorzata.Warmuzek wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } } I have a problem with an import the file with an extension *.sp ( spectrum ) } and put it to the document in word 6 or amipro } } Malgorzata Warmuzek } Foundry Research Institute } Research Materials Department } Structural and Physical Research Laboratory } } Zakopianska 73 Call +48 12 2605022 ext. 317 } 30-418 KRAKOW - POLAND Fax +48 12 2665478, +48 12 2660870 } } }
I don't have access to a nice workshop, where they can construct a glow discharger, where in Europe can I buy one? Or, is it out there someone, who has an old mashine you don't need anymore?
TIA
Gunnel Karlsson
^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ Gunnel Karlsson E-mail Gunnel.Karlsson-at-oorg2.lth.se Biomicroscopy Unit Tel +46 222 8229 Inorganic Chemistry 2 Fax +46 222 4012 Box 124 S-221 00 LUND, Sweden ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ This message was sent by Eudora with recycled electrons
Malgorzata, There should be some function on your Link system for outputting the data of a *.sp file onto a DOS disk. This can then be read in to Microsoft Excel, plotted as a graph and the graph imported into MS word. We have a different Link system to you so the details may vary on how you get the Link system to output onto a DOS disk but I would be happy to send you a copy of the procedure that works for us if that would help.
Yours sincerely
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
I have your book as a Bible on my desk. I remember our discussions concerning this problem as well but I do hope that I will find someone around the world who get closer to solve it. May be you know or heard about such person?
In cryofracturing we will be not able to obtain one layer of cells, that is a reason why I was trying cryo-sectioning. Freeze-substitution can be an option, but one should be careful about redistribution of ions which is our main worry in preparation for X-ray microanalysis.
Best regards
Jolanta
} As far a s cryosections of leaves are concerned. It ain't easy because } of all the internal air spaces. Cryofracturing is no problem and with a } kittle care you can cryoplane the leaf ti get a nice smooth surface. I } think I know what you want to do Jolanta (are you still doing ion beam } microscopy ?) May be freeze substitution is the best approach. If all } else fails read my book } } Best wishes } } Patrick Echlin } University of Cambridge
************************************************************************ Dr Jolanta Mesjasz-Przybylowicz National Accelerator Centre P.O. Box 72 Faure 7131 South Africa tel: 27-21-8433820 fax: 27-21-8433543 Internet: MESJASZ-at-nac.ac.za ************************************************************************
For that matter, if you are running under Windows, you can use the PrintScreen or Alt-PrintScreen keys to capture the screen and paste it into Word or whereever. You can get fancier by using the Format Picture item in Word to trim the bitmap to the area you want and to size it. Or you can get fancier still and first paste the bitmap into a picture editor (e.g., LView Pro or MS Imager or even MS Paint which come with Windows). Then you can crop or annotate the image and save it to a file or copy it from there to your word processor.
Note that the difference between PrintScreen and Alt-PrintScreen is that the Alt form copies only the active window instead of the whole screen. Thus you can size your spectrum (or any other application) as you want before snapping a copy.
This may not be as nice as importing the spectrum in a form in all its detail, but it will convey the information.
At 02:40 PM 9/25/97 +1000, you wrote: } Malgorzata, } } You can capture the spectrum with a software, such as Paint Shop } Pro we are using, and save it as TIFF or Bitmap file. If you have the } software, I can show you the details. } } With the best wishes, } Charlie Kong } kong-at-t-rex.materials.unsw.edu.au } } } On Wed, 24 Sep 1997, Malgorzata.Warmuzek wrote: } } } I have a problem with an import the file with an extension *.sp ( spectrum ) } } and put it to the document in word 6 or amipro } } } } Malgorzata Warmuzek } } Foundry Research Institute } } Research Materials Department } } Structural and Physical Research Laboratory } } } } Zakopianska 73 Call +48 12 2605022 ext. 317 } } 30-418 KRAKOW - POLAND Fax +48 12 2665478, +48 12 2660870 ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering, or 270 Metals Development Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-3091
To make a glow discharge all you need is a poor vacuum ( ~ 100 mTorr) and a high voltage power supply. You can use a vacuum bell jar with an electrical feedthru. Install a leak value in the system and rough pump it out, then put in a controlled leaked of what ever gas you want, air will work, but Argon is nice and be careful with Oxygen.
Arrange your electrode to be in the bell jar (insultate it up to the point where you want the "glow" to start.) Then slowly crank up the kV. Depending on your geometry, gas pressure etc.. you should get something by the time you reach ~ 1 kV.
The more important question is what do you want to do with the discharge?
Have you tried the (Walter) McCrone Institute in Chicago? I don't have the address handy, but maybe someone else does. They have worked on everything from the Shroud of Turin to ancient paints.
------------------------------------------------ Opinions or statements expressed herein, rational or otherwise ;-) do not necessarily reflect those of my employer.
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 Phone: (508) 750-1717 E-mail: crossman-at-osi.sylvania.com
Our web sites: www.sylvania.com www.siemens.com --
Greetings All, After each step of the UA stain and the LC stain, I rinse the copper grids by immersion. I have four changes of water and "dip" the grid about 40 times in each change. I am interested in changing that to the procedure that rinses the grid by "flooding" it using a syringe or whatever. Can someone share with me exactly how that is done ... what type of water ... etc. Also ... do the sections ever wash off the grid when rinsing that way??
Thanks in advance, Sharron Chism HT (ASCP) Electron Microscopy Lab Harris Methodist Hospital Fort Worth, Texas
09/25/97 Hi everywhere/everyone, as a brand-new member of this listservice I want to add a notice on = self-fabrication of silicone rubber molds for epoxide-resin-embedding, = which Lonie Kerr asked for 10/28/96 (only for the case, someone has = similar problems now). There are articles (in English) on self fabrication of moulds for = epoxide-embedding, at least 2 of them are: 1) MUSS W., SIMONSBERGER P. (1983): Home-made Silicone Rubber Embedding = moulds for Electron Microscopy, Mikroskopie (Vienna) 40, 207-209 (simple version for the unexperienced) 2) MUSS W.(1984): Self fabrication of Silicone Rubber embedding Moulds = for use in Electron Microscopy; Mikroskopie (Vienna) 41, 34-39 (sophisticated version for the unexperienced including silicone rubber = material to be used). I am producing all my moulds (types as shown in article 2) like cubic, = flat, "beem capsule"- type w/round or pyramidal block face, with = engraved numbers for identification and others as requested) by myself. = They are needed in our routine diagnostic EM-Lab and fabricated since = 1984 without problems, with high quality in shearing force and = unmoulding properties as well as very low costs (quantity needed a year = for about 1300 to 1600 specimen blocks ~ 5 moulds 48 specimen = cabinets/each; costs/mould ~ US-Dollars 5-15.-, depending on amount of = silicone rubber mass needed). Fabrication is simple, if negative moulds of sufficient quality are at = hand and several general rules in working up the silicone mass are = strictly followed.=20 Silicone rubber moulds of this type are o.k. for use with epoxide = resins, use for hydrophilic resins like LR White, Lowicryls not tested = yet. If interested in how to produce such moulds efficiently and interested = in which kind/quality of silicone rubber one should use, please send = e-mail request to:
Wolfgang MUSS PhD Head of EM-Lab at Pathology Department LKA Muellner Hauptstrasse 48 A-5020 SALZBURG/AUSTRIA/Europe phone: ++43++662+4482-4720 Ext. fax: ++43++662+4482-882 Ext (c/o W. MUSS) e-mail: W.Muss-at-lkasbg.gv.at.
Good luck, hope this helps somebody. END of Message, no attachment added.
} I don't have access to a nice workshop, where they can construct a glow } discharger, where in Europe can I buy one? Or, is it out there someone, who } has an old mashine you don't need anymore? } } TIA } } Gunnel Karlsson
You don't need a workshop to make a usable system. Get a small, cheap handheld Tesla coil of the sort used for physics demonstrations. Identify an unused current feedthru in your vacuum evaporator, and attach a wire fitting to it to use as a "docking port" for the Tesla coil. Discharge the coil into the vacuum during rough (mechanical) pump. Take care to not discharge the coil in air; the ozone produced damages the cilia in your respiratory system.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
Dear Randy, } } We are looking for any information we can find on the } microscopy/identification of particles from automobile tires. Seems we } have a case where we need to try to match particles from a pair of tennis } shoes to a particular set of automobile tires(best case), or at least } identify particles as possibly coming from automobile tires (more likely). } (I'll let your imagination fill in the details on this one!) } } If anybody has done anything like this, please let us know. In the } meantime, we'll be hitting the indexes/abstracts and databases. } We have looked at some polymer blends for which the components can be differentially stained, and I can give you the name of our user who could possibly tell you whether this can be done for tire particles. The HVEM can be used to look at the structures of such particles if they are some few micrometers thick. We can also do EDX (and have done so on a few forensic specimens). Yours, Bill Tivol
See the following article for an easy to build glow discharge unit. I built a similar one several years ago for ionizing grids, and it works very well. All you need is a drill, a hack saw, a drill bit, and some epoxy. You will also need a plastic desciccator, a few feet of wire, a few machine and sheet metal screws, and some sheet aluminum about 1/8 to 1/16 of an inch thick. Mine is simpler than the one described in the article, consisting of only an aluminum disk screwed into the cut-off end of the high voltage generator, a larger disk placed inside the desciccator (with a wire going to an outside ground). You will also need a mechanical vacuum pump.
Aebi U, 1987 [See Related Articles] A glow discharge unit to render electron microscope grids and other surfaces hydrophilic. J Electron Microsc Tech 7(1), 29-33 (1987) ---------------------- Doug Keene DRK-at-shcc.org
See the following article for an easy to build glow discharge unit. I built a similar one several years ago for ionizing grids, and it works very well. All you need is a drill, a hack saw, a drill bit, and some epoxy. You will also need a plastic desciccator, a few feet of wire, a few machine and sheet metal screws, and some sheet aluminum about 1/8 to 1/16 of an inch thick. Mine is simpler than the one described in the article, consisting of only an aluminum disk screwed into the cut-off end of the high voltage generator, a larger disk placed inside the desciccator (with a wire going to an outside ground). You will also need a mechanical vacuum pump.
Aebi U, 1987 [See Related Articles] A glow discharge unit to render electron microscope grids and other surfaces hydrophilic. J Electron Microsc Tech 7(1), 29-33 (1987) ---------------------- Doug Keene DRK-at-shcc.org
Wolfgang: I , for one, am very interested in learning more about this technique. I started trying to do this last month without a lot of success. I just looked for your publication but unfortunately our somewhat mediocre library doesn't carry Mikroskopie . So it would be a great help to me if you could outline your procedure. I am especially interested in your formulation of silicon rubber and where you buy the components. Thanks in advance. Tom Phillips
-------------------------------------. } } 09/25/97 } Hi everywhere/everyone, } as a brand-new member of this listservice I want to add a notice on } self-fabrication of silicone rubber molds for epoxide-resin-embedding, } which Lonie Kerr asked for 10/28/96 (only for the case, someone has } similar problems now). } There are articles (in English) on self fabrication of moulds for } epoxide-embedding, at least 2 of them are: } 1) MUSS W., SIMONSBERGER P. (1983): Home-made Silicone Rubber Embedding } moulds for Electron Microscopy, Mikroskopie (Vienna) 40, 207-209 } (simple version for the unexperienced) } 2) MUSS W.(1984): Self fabrication of Silicone Rubber embedding Moulds for } use in Electron Microscopy; Mikroskopie (Vienna) 41, 34-39 } (sophisticated version for the unexperienced including silicone rubber } material to be used). } I am producing all my moulds (types as shown in article 2) like cubic, } flat, "beem capsule"- type w/round or pyramidal block face, with engraved } numbers for identification and others as requested) by myself. They are } needed in our routine diagnostic EM-Lab and fabricated since 1984 without } problems, with high quality in shearing force and unmoulding properties as } well as very low costs (quantity needed a year for about 1300 to 1600 } specimen blocks ~ 5 moulds 48 specimen cabinets/each; costs/mould ~ } US-Dollars 5-15.-, depending on amount of silicone rubber mass needed). } Fabrication is simple, if negative moulds of sufficient quality are at } hand and several general rules in working up the silicone mass are } strictly followed. } Silicone rubber moulds of this type are o.k. for use with epoxide resins, } use for hydrophilic resins like LR White, Lowicryls not tested yet. } If interested in how to produce such moulds efficiently and interested in } which kind/quality of silicone rubber one should use, please send e-mail } request to: } } Wolfgang MUSS PhD } Head of EM-Lab at Pathology Department LKA } Muellner Hauptstrasse 48 } A-5020 SALZBURG/AUSTRIA/Europe } phone: ++43++662+4482-4720 Ext. } fax: ++43++662+4482-882 Ext (c/o W. MUSS) } e-mail: W.Muss-at-lkasbg.gv.at. } } Good luck, hope this helps somebody. } END of Message, no attachment added.
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
I published an article entiled as "A Modified Autowasher Device for Rapidly Washing Large Numbers of EM-grids" in Microscopy Researdh and Technique vol 26:177-179 (1993). Here I just copy the summary part for you:
This device consists of a siphon system and 5 or 10 grid disks, modified from the previous mode (Chen, 1973), for large numbers of grids with ultrathin sections. This method improves the ease of assembling grids onto the grids disk and also requires much less stain solution. This system only takes 5 min for one single stain washing, at a maximum of 100 grids, and also avoids stain contamination. The grid disk can also be used for immunocytochemistry work and for critical point drying of grids with biological specimens.
You may go to SPI Supplies website at http://www.cccbi.chester.pa.us/spi/new/stanwash.html/ for details.
Good luck,
Ming
On Wed, 24 Sep 1997, Chism, Sharron wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Greetings All, } After each step of the UA stain and the LC stain, I rinse the } copper grids by immersion. I have four changes of water and "dip" the } grid about 40 times in each change. I am interested in changing that to } the procedure that rinses the grid by "flooding" it using a syringe or } whatever. Can someone share with me exactly how that is done ... what } type of water ... etc. Also ... do the sections ever wash off the grid } when rinsing that way?? } } Thanks in advance, } Sharron Chism HT (ASCP) } Electron Microscopy Lab } Harris Methodist Hospital } Fort Worth, Texas }
*********************************************** * Ming H. Chen, PhD * * Medicine/Dentistry Electron Microscopy Unit * * University Of Alberta. * * Edmonton, Alberta, Canada * * * * Visit My Page At: * * http://www.ualberta.ca/~mingchen * ***********************************************
Mark, I've been using a product called imageAXS CE v3.0. This is a free version of a slightly more flexible commercial program. It automatically finds image files and creates a MS Access database which you can edit and use for searching and indexing images. the url is:
http://www.dascorp.com
Hope this helps glenn
Glenn Poirier Tel (514) 398 -6774 Electron Microprobe Laboratory Fax (514) 398 4680 Earth and Planetary Sciences glennp-at-stoner.eps.mcgill.ca McGill University http://castaing.eps.mcgill.ca 3450 University St. Montreal, Qc H3A 2A7
There are three sides to every story: Your side, My Side and the truth
Dear Tom, I have been casting my own silicon rubber molds for some time now. These are one-inch molds for epoxy metallurgical mounts. The silicon rubber comes from Dow Corning and they have several strengths and hardnesses. First, I have a kit consisting of several 100 ml tri-pour (plastic) beakers with a hole drilled in the bottom to fit a short screw. A block of aluminum, one inch in diameter and about 3/4 inch high, is tapped to receive the screw. You screw the aluminum block securely into the bottom of the beaker, mix the SiRubber (according to the directions) in a disposable cup, outgas the SiRubber in a vacuum desicator, then pour the rubber into the beaker. Outgas again. Leave it overnight to set, then remove the set rubber in the morning. It is a bit of a struggle to remove: I take out the screw, then push up on the Al block with a sharp point. Any flaps can be cut off with a razor blade. You can cast these things in any shape you can imagine.
You wrote: } Wolfgang: I , for one, am very interested in learning more about this } technique. I started trying to do this last month without a lot of } success. I just looked for your publication but unfortunately our somewhat } mediocre library doesn't carry Mikroskopie . So it would be a great help } to me if you could outline your procedure. I am especially interested in } your formulation of silicon rubber and where you buy the components. } Thanks in advance. Tom Phillips } } -------------------------------------. Regards, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
What are some of the problems associated with making a photograph of a single atom? and What methods are used for getting around this problem? please send response to :
We currently use Potssium ferrocyanide reduced OsO4 as our 'routine' post fixative, primarily for enhanced membrane staining. When it works, tissue stains up beautifully! However, every now and then, we get tissue which appears to not have infiltrated properly ater using this post fixative. Nine times out of ten, it is a result of the potassium ferrocyanide (as repeat tests with new/no Pot Ferro show). According to the literature, (I think even to the original pot ferro paper) they do mention that extra care must be taken when infiltrating into epoxy resin when using this stuff. It only seems to be intermittent, even with fresh Pot ferro made up.
Does anybody know what is the pot ferro stock solution shelf life is? (we use stuff between 4 and 8 weeks old after 'brewing' for the first four) What does the pot ferro react with in the tissue to prevent good infiltration? Any other ideas/suggestions?
I'd appreciate any help on this,
Thanks
Rich.
P.S. Thanks for all the replies to an earlier request (4 - 5 weeks!) for info re: charging policies for EM Units too! :-)
----------------------------------------------------------------------- Richard Lander Electron Microscope Technician South Campus Electron Microscope Unit Otago School of Medical Sciences P.O. Box 913 Dunedin New Zealand. Tel. National 03 479 7301 Fax. National 03 479 7254
"Southernmost EM Unit in the World!" ------------------------------------------------------------------------
Hello all, Has anyone been successful in labelling Substance-p for TEM? I am trying to tag it in endothelial cells, and have been having as much luck as a snowball would have in hell. Any help would be appreciated, as the old PhD may hinge on this some day! While I am here, if anyone has used DiI (3,3'-dioctadecyloxacarbocyanine percholate) or DiO (1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine percholate) for neuronal tract tracing, please email me? ...................................................................... ....................................... Alex Black Department of Anatomy National University of Ireland Galway
alexander.black-at-ucg.ie
Alex Black BSc, MMedSc Department of Anatomy National University of Ireland, Galway Galway, Ireland
RENAL BIOPSY - IMMUNOFLUORESCENCE CONTROLS } Anyone doing immunofluorescenst techniques in renal biopsies running a control? } No.
One reason is that there is not enough tissue to go around. It is very difficult to get control sections from a positive case and there are not enough positive cases that are archived to pproduce the material.
Any suggestions? |--------------------------------| | Karlene Hewan-Lowe, M.B., B.S. | | Department of Pathology | | Emory University | | Phone: 404-686-2926 | | Fax: 404-6864978 | |--------------------------------|
Dear colleagues, thank you for replies to my topic from yesterday and your interest. = Lucky to see that there are some out there in need for such a procedure. = God bless you! To handle the problem via e-mail I think would go to far, concerning = spaceneeds of informations. Unfortunately I do have neither a homepage = (for the future there is planned one) nor a scanner unit, so I am not = able to reproduce those papers for you by e-mail. So I shall send ASAP = to all of you (see below) written infos to you. Unfortunately I don=B4t have original reprints of that papers mentioned = in my information.So you will get copies, as an attachment I shall send = Instructions which summarize the most important things in my opinion to = do the job as optimal as possible.=20 So my message to all who responded till now and others maybe following: Thank you all very much for your interest/greetings (Keith! welcome = greetings too!): - Jerzy BOHDANOWICZ, GDANSK/Poland, - Julian P.S. SMITH III, ROCK-HILL, SC/USA - Phil OSHEL, CHAMPAIGN, IL/USA (will contact you separately with = respect to "Microscopy today") - Tom PHILLIPS, COLUMBIA, MO/USA - Ann Fook YANG, OTTAWA, ONT/CANADA - Scott WHITTAKER, GAINESVILLE, FL/USA - Gabriel Adriano ROSA, BUENOS AIRES/ARGENTINA=20 - John J. BOZZOLA, CARBONDALE, IL/USA you will get written/copied information ASAP. Sincerely yours Wolfgang MUSS Dept. Pathology LKA (Gen.County Hospital), EM-Lab, Muellner Hauptstrasse = 48, A-5020 SALZBURG/AUSTRIA-Europe, phone: ++43++662+4482+4720 Ext, = Fax:++43++662+4482+882 Ext ("c/o W.MUSS") End of message, no attachment added.
Re to Sharron CHISM, Fort Worth Tx; 09/26/97 Dear Sharron, whatever type of aid (apparatus) you will use for staining your = ultrathins: concerning water quality: if possible, use UHQ (ultrahigh-purified = water) or at least bi- (better triple-)distilled water from a = quartz-glass distilling apparatus (most conveniently in your own lab!). = My/our story: our quartz-glass bi-distilling apparatus which at that time had a = life-span of nearly 12 years (in which time we got no bigger problems at = all) had broken down (heating wire burned through). Therefore we thought = to overcome the problem in ordering } } A.bidest.sterile, "non pyrogenic" = { { via our hospital pharmacy (+/- every time freshly produced, etc.). In = fact we got worsest stainings of our sections due to precipitates never = seen before in such an amount and shapes. We thought this to be a = possible source with respect to our handling in washing the grids, = unclean glass ware, syringes, etc.... or just more dust in the lab or = else; nothing of that all: despite using same methods for mixing up our = staining solutions as usual, we were not able to locate this source of = junk on the grids! When I asked at our pharmacy, how they produce their = water, they showed me & told me about their apparatus: it was/is a still = producing vapours by means of copper plates! An analytic chemist told me = some days after, that their central analytic lab got their own = distilling (UHQ) machine because of too high copper-contents of the = bidistilled water of the former source. Since that info I used only = their UHQ (coming along in clean glassware, most preferably quartz = glass) with no precipitate-problems any more, hoping to get my old = quartzglass still as soon as possible from repairing. Hope this adds another aspect in "hunting our elephantine precipitates", Best regards Wolfgang MUSS SALZBURG/Austria, Europe
Sharron, I also stain with Uranyl Acetate 5 min and Lead Citrate 5 min. I place copper grids, section side down, on drops of UA and LC. Inbetween stain and after I grasp edge of copper grid and flood by dripping from top of forcep for approx 10 sec each grid. I use Distilled, Sterile water to rinse. any reason for change. I know several techs that rinse as you do, but felt there needed to be changed. I've been staining as above for over 22 years. } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I would like to point out another source of contamination in stills: the tubing. Not the obvious crud, but breakdown products. We were using manufacturer-recommended Tygon tubing on our Barnsted still, and we getting some mysterious scum in the distillate. The micro-analytical lab couldn't identify it, other than as some complex organic kind of thing. Process of elimination ended at the tubing. The distillate was coming off at below, but not much below, the breakdown temperature of the tubing. The tubing was changed for higher-temperature rated silicone tubing. This seemed to solve the "new contamination" problem, but the previous contamination had coated the inside of the glass (not sure if it was quartz) carbouy and wouldn't clean off (not even will Tilex Scum Remover).
Phil } } Re to Sharron CHISM, Fort Worth Tx; 09/26/97 } Dear Sharron, } whatever type of aid (apparatus) you will use for staining your ultrathins: } concerning water quality: if possible, use UHQ (ultrahigh-purified water) } or at least bi- (better triple-)distilled water from a quartz-glass } distilling apparatus (most conveniently in your own lab!). My/our story: } our quartz-glass bi-distilling apparatus which at that time had a } life-span of nearly 12 years (in which time we got no bigger problems at } all) had broken down (heating wire burned through). Therefore we thought } to overcome the problem in ordering } } A.bidest.sterile, "non pyrogenic" { { } via our hospital pharmacy (+/- every time freshly produced, etc.). In fact } we got worsest stainings of our sections due to precipitates never seen } before in such an amount and shapes. We thought this to be a possible } source with respect to our handling in washing the grids, unclean glass } ware, syringes, etc.... or just more dust in the lab or else; nothing of } that all: despite using same methods for mixing up our staining solutions } as usual, we were not able to locate this source of junk on the grids! } When I asked at our pharmacy, how they produce their water, they showed me } & told me about their apparatus: it was/is a still producing vapours by } means of copper plates! An analytic chemist told me some days after, that } their central analytic lab got their own distilling (UHQ) machine because } of too high copper-contents of the bidistilled water of the former source. } Since that info I used only their UHQ (coming along in clean glassware, } most preferably quartz glass) with no precipitate-problems any more, } hoping to get my old quartzglass still as soon as possible from repairing. } Hope this adds another aspect in "hunting our elephantine precipitates", } Best regards } Wolfgang MUSS } SALZBURG/Austria, Europe
Among the awards to be presented next year is the "Morton D. Maser Distinguished Service Award," to be presented to an individual to ..."recognize outstanding volunteer service to the Society, ... and who has served the Society for many years with great dedication."
The nomination is to include a letter (this e-mail) from the primary nominator and supplemental letters (e-mails) of support from others.
It is with great personal pleasure that I nominate Nestor Zaluzec for this award.
Nestor is our friendly SYSOP, has organized the Argonne e.m. computer resources on behalf of microscopists everywhere, has organized the computer workshop/software exchanges at our meetings, served as MSA Program Chair for the Minneapolis meeting, etc., etc.
If you would like to provide a supplemental e-mail of support please direct your contribution to Gracie Burke, MSA Awards Committee. Deadline for input is December 31, 1997.
mgburke-at-pitt.edu
Thank you.
Ron Anderson
p.s. Please DO NOT send or copy your letters to the listserver, it's bad enough that Gracie will never speak to me again!
What is probably happening is that the paraffin wax is seperating from the monomers that have been added to it. This can happen under several circunstances. The most common being that the temperature on the paraffin dispencer is set too high for the wax being used. Another reason could be that the wax has been sittting unused (but hot) for too long a time (several weeks or more).
-- Begin original message --
} From: Gary Radice {gradice-at-richmond.edu}
} } I keep Paraplast Plus melted in a bath more or less continuously, so it is } always ready to use, even though it may be one or two months between uses. } Lately my students have been having some sectioning problems (with freshly } sharpened blades) and before I suggest that maybe they have done something } wrong during embedding, is it possible that the wax has "gone bad?" } } Gary Radice, Associate Professor gradice-at-richmond.edu } Department of Biology 804-289-8107 (voice) } University of Richmond VA 23173 804-289-8233 (FAX) -- End original message --
regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
} } Malgorzata Warmuzek wrote: } } I have a problem with an import the file with an extension *.sp ( spectrum ) and put it to the } } document in word 6 or amipro
Dear Malgorzata,
I use the ISIS suite 3.0 and there is an easy way to copy spectra to any other windows program, maybe it works also with your version. Use the command 'Buttons' 'Print' and a new window will appear called 'Spectrum printing'. There it is possible to activate the command 'Edit' 'Copy'. The Spectrum will now be transfered to the clipboard in form of a vector graphic.
Kind regards
Rainer ------------------------------------------- Rainer Ziel Akzo Nobel Central Reasearch 63784 Obernburg - Germany
It's possible to make excellent moulds from the material that dentists use to make dental impressions. This is usually polyvinyl siloxane, though there is also a polyether material that is more awkward to work with. The polyviynl siloxane (Perfourm, Reprosil etc.) comes in tubes (cheaper) or in cartridges which give a much better result. We have been using these materials in our lab for many years. They are available from any dental supplier. Hope this helps. Lesley Weston.
On Thu, 25 Sep 1997, Wolfgang Muss wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } 09/25/97 } Hi everywhere/everyone, } as a brand-new member of this listservice I want to add a notice on self-fabrication of silicone rubber molds for epoxide-resin-embedding, which Lonie Kerr asked for 10/28/96 (only for the case, someone has similar problems now). } There are articles (in English) on self fabrication of moulds for epoxide-embedding, at least 2 of them are: } 1) MUSS W., SIMONSBERGER P. (1983): Home-made Silicone Rubber Embedding moulds for Electron Microscopy, Mikroskopie (Vienna) 40, 207-209 } (simple version for the unexperienced) } 2) MUSS W.(1984): Self fabrication of Silicone Rubber embedding Moulds for use in Electron Microscopy; Mikroskopie (Vienna) 41, 34-39 } (sophisticated version for the unexperienced including silicone rubber material to be used). } I am producing all my moulds (types as shown in article 2) like cubic, flat, "beem capsule"- type w/round or pyramidal block face, with engraved numbers for identification and others as requested) by myself. They are needed in our routine diagnostic EM- Lab and fabricated since 1984 without problems, with high quality in shearing force and unmoulding properties as well as very low costs (quantity needed a year for about 1300 to 1600 specimen blocks ~ 5 moulds 48 specimen cabinets/each; costs/mould ~ US- Dollars 5-15.-, depending on amount of silicone rubber mass needed). } Fabrication is simple, if negative moulds of sufficient quality are at hand and several general rules in working up the silicone mass are strictly followed. } Silicone rubber moulds of this type are o.k. for use with epoxide resins, use for hydrophilic resins like LR White, Lowicryls not tested yet. } If interested in how to produce such moulds efficiently and interested in which kind/quality of silicone rubber one should use, please send e-mail request to: } } Wolfgang MUSS PhD } Head of EM-Lab at Pathology Department LKA } Muellner Hauptstrasse 48 } A-5020 SALZBURG/AUSTRIA/Europe } phone: ++43++662+4482-4720 Ext. } fax: ++43++662+4482-882 Ext (c/o W. MUSS) } e-mail: W.Muss-at-lkasbg.gv.at. } } Good luck, hope this helps somebody. } END of Message, no attachment added. } }
We have a problem obtaining frozen ultra-thin sections of cultured cells. We have tried both 2.3 M sucrose and 20% PVP/1.8 M sucrose to infiltrate the cells before freezing without success. Briefly, after fixing the cells and rinsing with buffer, the plates are scraped and the cells are microfuged briefly to pellet the cells. Then we add a small amount (approximately 1X the volume of the cell pellet) of sucrose or PVP/sucrose to the pellet of cells, and tap the tube to resuspend the cells. The next day a small amount of the cell suspension is placed on a metal peg and frozen in liquid nitrogen. When sectioning we obtain snow-like flakes on the knife edge. Tissues processed in a similar manner cut well. We suspect the problem to be too much buffer associated with the cell pellet, even though we try to remove as much buffer from the pellet as possible. Any suggestions would be appreciated.
Christine Roy and David Hall Albert Einstein College of Medicine Bronx, NY
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Considerable! If you or anybody else can image a single atom in an SEM, then you're edging into Nobel Prize territory (although it's relatively easy in a dedicated STEM). Under the correct circumstances, it's not too difficult to do it in a TEM either. For example, isolated uranium atoms on a thin carbon film support are relatively easy. If you can manage a single sulphur atom on a carbon support in a SEM, then I guess you'll be off to Sweden. On the other hand, there are a number of 'probe' instruments - STM, AFM, etc - around which can visualise individual atoms without any difficulty. However, if you want to understand what the image really means, then it starts to get difficult:)
This seems to me to be another case of where the real answer to the question is a visit to your local library. I know they aren't necessarily 'high-tech', but there still isn't much around electronicaly to beat a visit to a good library, sitting down, going through the journals, indexes, citation catalogues, etc. It takes time but it will give you what you want - refereed papers in reputable journals.
Sharron, if you don't like the dip method try the dilution method, because any flooding method with the syringe or whatever may destroy your sections, try allowing the grid to float to the bottom of a small beaker (10-20 ml) repeat 3-5 times, pour off the excess water to retrieve the grid, and rember to pick it up by the edge of the grid. -MR
On Wed, 24 Sep 1997, Chism, Sharron wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Greetings All, } After each step of the UA stain and the LC stain, I rinse the } copper grids by immersion. I have four changes of water and "dip" the } grid about 40 times in each change. I am interested in changing that to } the procedure that rinses the grid by "flooding" it using a syringe or } whatever. Can someone share with me exactly how that is done ... what } type of water ... etc. Also ... do the sections ever wash off the grid } when rinsing that way?? } } Thanks in advance, } Sharron Chism HT (ASCP) } Electron Microscopy Lab } Harris Methodist Hospital } Fort Worth, Texas }
Sharron, if you don't like the dip method try the dilution method, because any flooding method with the syringe or whatever may destroy your sections, try allowing the grid to float to the bottom of a small beaker (10-20 ml) repeat 3-5 times, pour off the excess water to retrieve the grid, and rember to pick it up by the edge of the grid. -MR
On Wed, 24 Sep 1997, Chism, Sharron wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Greetings All, } After each step of the UA stain and the LC stain, I rinse the } copper grids by immersion. I have four changes of water and "dip" the } grid about 40 times in each change. I am interested in changing that to } the procedure that rinses the grid by "flooding" it using a syringe or } whatever. Can someone share with me exactly how that is done ... what } type of water ... etc. Also ... do the sections ever wash off the grid } when rinsing that way?? } } Thanks in advance, } Sharron Chism HT (ASCP) } Electron Microscopy Lab } Harris Methodist Hospital } Fort Worth, Texas }
Hello, all- Are any of you interested in a Denton DFE-3 Freeze Etch Unit that fits on a Denton DV-502 vacuum evaporator? Please don't respond unless you are reasonably sure you know what this is, and think you might want it, anyway! It seems to be in excellent condition.
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
I am finally starting to prepare some TEM samples of glass. Does anyone know whether a light coat of carbon should be applied to both sides of the sample or is just one side (I assume the top side in the microscope) good enough?
-Scott
Scott D. Walck PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of S.D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
I have been asked to prepare some delicate fungal hyphae samples. I have a Denton critical point dryer and a freeze dryer. I'm more comfortable using the freeze dryer. Has anyone used a freeze drying technique on similar samples? Or does anyone in Alberta or British Columbia have a Denton critical point dryer that I might be able to come and see how to properly use. I'm not convinced that I'm using this one properly. Any suggestions appreciated.
It is indeed possible there may be excess buffer with the cells. Do you get *anything* in the way of sections that you can observe in the scope? If so, what do you see?
1. Do the fixed cells stay nicely as a pellet? After you fix and wash the cells, perhaps you could transitionally embed the pellet in low-melt agarose or 1.5% agar to hold the cells together. Do this only if the fixed pellet does not hold together on its own. Some fixed cells will form a nice pellet and stay that way. Others tend to go back into suspension.
2. *Definitely* increase the amount of cryoprotectant, well in excess of the volume of the pelletted cells. Try a trick which we used to use for some tissues: Assuming you have maybe a 50-100 microliter packed cell volume, fill a small 1-2 ml centrifuge tube, Eppendorf (TM) or similar, with the cryoprotectant. Then gently layer the cell pellet, embedded if necessary as in #1, above, on top of the cryoprotectant and very gently push the pellet *just under* the surface of the cryoprotectant. Allow the pellet to sink to the bottom of the tube on its own accord. This may take overnight. Infiltration with the cryoprotectant is complete when the pellet reaches the bottom of the tube. There is usually no need to leave the pellet in the cryoprotectant for longer periods of time. You can remove it and mount on a pin immediately after it reaches the bottom of the tube.
Good Luck! Let us know the secret after you get the problem solved.
Imaging atoms? Carefully consider the affects of potential probe size on the image, electron beam(and descrete potential electron optic subcomponents) and AFM tip size. SPM(AFM) and EM may be producing images heavily convoluted by tip or e-beam probe size contour. Atoms or multiple probe images?
---------- } From: Larry Stoter {LPS-at-teknesis.demon.co.uk} } To: mskittee-at-swbell.net; Microscopy-at-sparc5.microscopy.com } Subject: Re: SEM photography } Date: Friday, September 26, 1997 12:35 PM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } -----------------------------------------------------------------------. } } } } What are some of the problems associated with making a photograph of a } } single atom? and What methods are used for getting around this problem? } } please send response to : } } } } mskittee-at-swbell.net } } Considerable! If you or anybody else can image a single atom in an SEM, } then you're edging into Nobel Prize territory (although it's relatively } easy in a dedicated STEM). Under the correct circumstances, it's not too } difficult to do it in a TEM either. For example, isolated uranium atoms on } a thin carbon film support are relatively easy. If you can manage a single } sulphur atom on a carbon support in a SEM, then I guess you'll be off to } Sweden. On the other hand, there are a number of 'probe' instruments - STM, } AFM, etc - around which can visualise individual atoms without any } difficulty. However, if you want to understand what the image really means, } then it starts to get difficult:) } } This seems to me to be another case of where the real answer to the } question is a visit to your local library. I know they aren't necessarily } 'high-tech', but there still isn't much around electronicaly to beat a } visit to a good library, sitting down, going through the journals, indexes, } citation catalogues, etc. It takes time but it will give you what you want } - refereed papers in reputable journals. } } Regards, } Larry Stoter } } ------=_NextPart_000_01BCCADB.8B1B4500 Content-Type: text/html; charset=ISO-8859-1 Content-Transfer-Encoding: quoted-printable
{html} {head} {/head} {BODY bgcolor=3D"#FFFFFF"} {p} {font size=3D2 = color=3D"#000000" face=3D"Arial"} Imaging atoms? Carefully consider the = affects of potential probe size on the image, electron beam(and descrete = potential electron optic subcomponents) and AFM tip size. SPM(AFM) = and EM may be producing images heavily convoluted by tip or e-beam = probe size contour. Atoms or multiple probe images? {br} {font size=3D2 = color=3D"#008080"} {br} {br} {font color=3D"#000000"} ---------- {br} > =
} } I am finally starting to prepare some TEM samples of glass. Does anyone } know whether a light coat of carbon should be applied to both sides of the } sample or is just one side (I assume the top side in the microscope) good } enough? } -Scott
One side carbon coating is sufficient. It can be on the top or the down side in the microscope, though you should get a better image if you place it on top, actually. If you do not coat, be sure you will charge and explode the sample immediately.
} Date: Fri, 26 Sep 1997 15:16:43 -0400 } From: Dr. David Hall {hall-at-aecom.yu.edu} } To: Microscopy Forum {microscopy-at-sparc5.microscopy.com} } Subject: frozen thin sections of isolated cells } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } We have a problem obtaining frozen ultra-thin sections of cultured cells. } We have tried both 2.3 M sucrose and 20% PVP/1.8 M sucrose to infiltrate } the cells before freezing without success. Briefly, after fixing the cells } and rinsing with buffer, the plates are scraped and the cells are } microfuged briefly to pellet the cells. Then we add a small amount } (approximately 1X the volume of the cell pellet) of sucrose or PVP/sucrose } to the pellet of cells, and tap the tube to resuspend the cells. The next } day a small amount of the cell suspension is placed on a metal peg and } frozen in liquid nitrogen. When sectioning we obtain snow-like flakes on } the knife edge. Tissues processed in a similar manner cut well. We } suspect the problem to be too much buffer associated with the cell pellet, } even though we try to remove as much buffer from the pellet as possible. } Any suggestions would be appreciated. } } Christine Roy and David Hall } Albert Einstein College of Medicine } Bronx, NY } Sounds to me as if your sections are sucrose (the "snow" or white stuff). I suspect your cells are thinly dispersed in the sucrose, and not stuck together enough to make a "section."
We add paraformaldehyde to the monolayers, swirl for just a few min (~5), scrape with a rubber policeman and pellet into a small tipped tube:
| | | | | | | | | | | | | | | | | | | | \ / H U almost exactly this size.
We pellet in a swinging bucket centrifuge and then microfuge to pack the cells. We let them fix for another hour or two and cut off the very bottom and again just above the cells, forming a log with the cells in the center that can be pushed out with a paper clip. If they stick together, fine, proceed. If not, push them into small piles (~0.5-1 mm), on a piece of Parafilm, drain them with filter paper cut into pie-shaped wedges using the very tip to touch the pellet gently, and coat them with cooled, still molter 1% agar. Cut away any excess agar. Inflitrate with 3 changes of sucrose (2.3M) over about 30-60 min. Place onto stubs and flash freeze. This keeps the cells together, not dispersed thinly in your sucrose.
If you fix very long before pelleting, your cells will not like to stick together, and will disperse in the sucrose. The consistency of the cell pellet should be like cooked oatmeal. (I could make some "snotty" comment about consistencies of other substances). If they are too wet, they will disperse, and you'll have to hunt all over your grid for them. If they're too dry, you could alter the ultrastructure.
Good luck. S
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Below is the result of your feedback form. It was submitted by (dmatthew-at-providence.edu) on Saturday, September 27, 1997 at 16:01:50 ---------------------------------------------------------------------------
Email: dmatthew-at-providence.edu Name: Douglas Matthews
School: Providence College
State: RI Question: Can anyone help me find a recipe for Millonig's phosphate buffer?
All MSA Members are reminded that nominations are currently being solicited for the 1998 MSA Awards. The Awards include:
MSA Distinguished Scientist (Biological Sciences) MSA Distinguished Scientist (Physical Sciences) MSA Burton Medal MSA Outstanding Technologist (Biological Sciences) MSA Outstanding Technologist (Physical Sciences) MSA Morton D. Maser Distinguished Service Award
Distinguished Scientist Awards: These Awards honor preeminent scientists from both the Biological and Physical disciplines who have an internationally recognized record of outstanding achievements in the field of microscopy and microanalysis.
Burton Medal: The Burton Medal was initiated to honor the distinguished contributions to the field of microscopy and microanalysis of a scientist who is not older than 35 years of age in the January of the award year.
Outstanding Technologist Awards: These Awards honor technologists from both the Biological and Physical Sciences who have made significant contributions such as the development of new techniques which have contributed to the advancement of microscopy and microanalysis.
Morton D. Maser Distinguished Service Award: This Award was initiated to recognize outstanding volunteer service to the Society as exemplified by Mort Maser, who served the Society for many years with great dedication. This award is made to honor an MSA member who has provided significant volunteer service to the Society over a period of years.
The Distinguished Scientist, Burton Medal, and Outstanding Technologist Awards Nominations should include: 1) a letter from the primary MSA nominator describing the research accomplishments of the candidate with particular emphasis on the unique technical achievements in the Physical or Biological Sciences; and 2) supplemental letters of support from other members of the scientific community.
The Morton D. Maser Distinguished Service Award Nomination should include: 1) a letter from the primary MSA nominator describing the basis for the nomination; and 2) supplemental letters of support from other members of MSA.
The Deadline for receipt of Awards Nomination Packages is December 31, 1997. Please contact the MSA Awards Committee [Gracie Burke (mgburke-at-pitt.edu), Stan Erlandsen (stan-at-lenti.med.umn.edu), Bev Maleef (Beverly_E_Maleeff-at-sbphrd.com), Jim Bentley (bentleyj-at-ornl.gov), Bill Gunning (wgunning-at-magnum.mco.edu) ] or the MSA Business Office for additional information. NOTE: E-mail "letters" are acceptable!
Thanks!
Gracie Burke
Dr. M.G. Burke Westinghouse Electric Corp. Bettis Laboratory West Mifflin, PA 15122 tel: (412) 476-5883; fax: (412) 476-5151 e-mail: mgburke-at-pitt.edu
Dear Douglas, 09/29/97 original recipe for MILLONIG=B4s buffer was published as follows: MILLONIG G: Advantages of a Phosphate Buffer for OsO4-Solutions in = Fixation. Abstract B 26, publ. on behalf of the EMSA in: Journal of Applied Physics (LANCASTER =3D Procs of the ann. Meetings of = EMSA) 32, p.1637 (1961) Original Text of this abstract:
} } Since the sodium-mono- and diphosphate exists as an effective buffer = system in the body fluids of animals, it has seemed reasonable to test = it as a vehicle for OsO4 in fixation of biological tissues. An isotonic = (delta=3D -0.56 degrees centigrade) solution at pH 7.3 of the following = composition was tested: sol. a) 2,26% NaH2PO4 sol. b) 2.52% NaOH sol. c) 5.4% Glucose (about 5 ml added to an end volume of OsO4-soln. = of 50 ml.)
[sol. d): 41,5 ml sol. a) + 8.5 ml sol. b) [fixative: 45 ml sol. d) + 5 ml sol. c) + 0.5g OsO4 (as crystals)] [It has been found that this solution is stable for several weeks, if = stored in the refrigerator in a clean bottle. In comparative studies, = mainly with the veronal-acetate buffer + Osmium and on different animal = tissuies, the phosphate buffer seems to prevent extraction of the = cytoplasmic matrix, preserves the glycogen and fibrillar elements and = gives an uniform fixation at different levels in the tissue block { {] --------End of original abstract text of Millonig himself.---------
Recipe for convenient lab-volumes therefore: for an isotonic buffer solution ( ~ 300 mosmol, pH ~ 7.2 - 7.3 ) u = should mix the following: 5.65 g NaH2PO4.H2O ad 250 ml of A.dest ( =3D 33.9 g ad 1500 ml) 1.26 g NaOH-pastilles ad 51 ml A.bidest ( =3D 7.76 g ad 307 ml) makes: ~300 ml ( ~ 1800 ml)
You should measure pH, as well as osmolality: in my experience osmolality is a little bit lower than 300 mosmols therefore I have to add about 0.1 to 0.2 (w/v) of D-Glucose (~0.5 g, or = ~3.3g respectively, for endvolume of 1800 ml) to end up with 300 = mosmols.
MILLONIG=B4s 0.13 M sodium phosphate buffer (isoosmotic with mammalian = blood) (from: MILLONIG G. (ed): Laboratory Manual of Biological Electron = Microscopy; published and distributed by Mario SAVIOLO-Editore, = VERCELLI/ITALIA, copyright by G.Millonig, 1976 , 67 pages) Solution A: 0.164 M monosodium phosphate ( =3D 2.26% NaH2PO4.H2O, =3D 2.56% NaH2PO4 .2H2O) Solution B: 0.63 N sodium hydroxide (=3D 2.52% NaOH)
pH 6.0 6.2 6.4 6.8 7.0 7.2 = 7.4 7.6 7.8 ml A 96.2 94.7 92.5 87.9 85.8 83.9 82.5 = 81.6 80.8 ml B 3.8 5.3 7.5 12.1 14.2 16.1 = 17.5 18.4 19.2=20
The Centre for Microscopy and Microanalysis at the University of Queensland has the following specimen holders surplus. All holders to be sold in working order, the cold stage controller is not included. Interested parties should contact,
Assoc. Prof. Alasdair W. McDowall Deputy Director: Center for Microscopy & Microanalysis University of Queensland, Brisbane. QLD 4072 Phone: (07) 3365-4211 International: 61 (7) 3365-4211 Facsimile: (07) 3365-4422 International: 61 (7) 3365-4422 WWW: http://www.uq.oz.au/nanoworld/nanohome.html
EXCESS SPECIMEN HOLDERS DUE TO JEOL 4000FX-4010 UPGRADE JEOL 4000FX SPECIMEN HOLDERS (a) GATAN (D8020106-1) Normal double tilt, motorised, low background. Holder has had routine use. (b) GATAN (D8020105) Tilt holder, motorised rotation. Holder has not been used. (c) JEOL standard single tilt holder. Holder has had minimum use. Also Hitachi 200kV TEM H 800 Specimen Holder (d) GATAN ( D003130 ) LN2 holder. Holder has had minimum use.
Cost: Aus$ 5,000.00 per holder plus shipping.
-- ************************************************************ Duncan Waddell (BSc) Centre for Microscopy and Microanalysis The University of Queensland. St. Lucia. Qld. 4072 Telephone: +61-7-3365-4216 Facsimile: +61-7-3365-2199 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. ************************************************************
Following the thread about coating of a glass sample, Jacky Larnould added off line that when the objective aperture is set (above the sample) the "charging effect" disappears, and the sample does no break. While I have noticed this phenomenon for a while I have never come with a satisfactory explanation. Has anyone found a good explanation to this?
} I am finally starting to prepare some TEM samples of glass. Does anyone } know whether a light coat of carbon should be applied to both sides of the } sample or is just one side (I assume the top side in the microscope) good } enough? }
We only ever coat one side. We usually try to put the coated side towards the electron source, but not always. It has always worked fine in preventing charging. I've often wondered why this should be, but it certainly works, on either ceramics or polymers!
Tony.
Anthony J. Garratt-Reed MIT Room 13-1027 77 Massachusetts Avenue Cambridge, MA 02139-4307 United States of America
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Reply to: RE} frozen thin sections of isolated cells
Dear Christine and David, We routinely prepare culture cell pellets for ultra-thin cryosectioning but we tend to put the resuspend the cells in gelatin then pellet this to form a nice gelled block of cells which can be treated the same as tissue for cryosectioning. Here is a copy of our protocol. If you have any questions please feel free to call me at 203-785-3646. You may also want to try adjusting the temperature and thickness you are cutting your cells. This may alleviate the "snow" effect. Linda Chicoine Center for Cell Imaging yale Univ. School of Medicine linda.iadarola-at-yale.edu EMBEDDING CELLS IN GELATIN/SUCROSE
1. Fix cells in buffered fixative directly in the culture dish. Can fix for 30' to overnight at 4C.
2. Remove fixative and replace with PBS/10%FCS to cover monolayer.
3. Scrape cells off culture dish with teflon , use glass pippette to transfer cells to an eppendorf tube.
4. Centrifuge gently to form a loose pellet.
5. Remove half the PBS/FCS and replace with 10% gelatin at a 1:1 dilution with PBS/FCS still in the tube. Final concentration 5% gelatin. Resuspend cells in this and re-centrifuge gently to form a pellet again.
6. Place tube in ice bucket for 15-30' or in fridge for longer, until gelatin has hardened.
7. Cut bottom of tube with a razor blade. Cut straight through tube to get the gelled pellet in the bottom piece of the tube.
8. In a petri dish of buffer or sucrose sitting on ice, remove the cell pellet from the bottom of the tube using a shaved wooden stick. Under a dissecting microscope, cut the pellet into small triangular or rectangular pieces.
9. Place peices in eppendorf tube filled with sucrose. Leave pieces for 30' or longer to infiltrate.
10. Pour sucrose into small petri dish, on ice,under disecting microscope. Remove a piece using capillary action of a fine point forceps and place the piece on a specimen nail. Check for excess sucrose around the pellet. Remove excess sucrose with a wedge of filter paper.
11. Immediately plunge the nail with the specimen into liquid nitrogen. Swirl the specimen somewhat vigorously and when completely frozen place the specimen in a nunc tube on an aluminum storage cane. Specimens can remain on the cane in liquid nitrogen for several years.
--------------------------------------
We have a problem obtaining frozen ultra-thin sections of cultured cells. We have tried both 2.3 M sucrose and 20% PVP/1.8 M sucrose to infiltrate the cells before freezing without success. Briefly, after fixing the cells and rinsing with buffer, the plates are scraped and the cells are microfuged briefly to pellet the cells. Then we add a small amount (approximately 1X the volume of the cell pellet) of sucrose or PVP/sucrose to the pellet of cells, and tap the tube to resuspend the cells. The next day a small amount of the cell suspension is placed on a metal peg and frozen in liquid nitrogen. When sectioning we obtain snow-like flakes on the knife edge. Tissues processed in a similar manner cut well. We suspect the problem to be too much buffer associated with the cell pellet, even though we try to remove as much buffer from the pellet as possible. Any suggestions would be appreciated.
Christine Roy and David Hall Albert Einstein College of Medicine Bronx, NY
------------------ RFC822 Header Follows ------------------ Received: by QuickMail.Yale.edu with SMTP;26 Sep 1997 16:51:50 -0400 Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id OAA17617 for dist-Microscopy; Fri, 26 Sep 1997 14:09:43 -0500 Received: from post.aecom.yu.edu (post.aecom.yu.edu [129.98.1.4]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id OAA17613 for {microscopy-at-Sparc5.Microscopy.Com} ; Fri, 26 Sep 1997 14:09:42 -0500 Received: from worm.aecom.yu.edu (worm.aecom.yu.edu [129.98.90.160]) by post.aecom.yu.edu (8.8.7/8.8.7) with SMTP id PAA11598 for {microscopy-at-Sparc5.Microscopy.Com} ; Fri, 26 Sep 1997 15:18:49 -0400 (EDT) Message-Id: {3.0.1.32.19970926151643.006af944-at-mailserver.aecom.yu.edu} X-Sender: hall-at-mailserver.aecom.yu.edu (Unverified) X-Mailer: Windows Eudora Light Version 3.0.1 (32)
Hi Folks !
Does anyone of you know of some specific staining technique for Titanium, and where to find a cook-book for it? We need it for recovery of the new type of Gun Shot Residues.
Best regards Bengt Bengt Stocklassa , Managing Director Cox Analytical Systems AB | Phone: +46 31 7725300 House of Innovations, CTH | Fax: +46 31 7725600 412 96 - Gothenburg, SWEDEN | E-mail: bengt-at-xco.se
Hi Jill, I've done some preliminary work using Leica's FD unit on fungal material. I've FD Bergamot (Monarda didyma) leaves with a powdery mildew fungal infection and yeast (Sacc. cer). The leaves came out fine but the fungal hyphae were collapsed. The yeast came out very nicely. I ran both fixed (Karnovsky's) and unfixed pieces. All were cryo prepped by propane plunge freezing. All were examined at 2.0 kV uncoated. Overall I was pleased with the results, I haven't had a chance to go over all of the samples but the chemical fixation seemed to be better for maintaining the fungus. I would run both unfixed and fixed as I believe the mechanical agitation of adding a liquid solution may have washed away some fungal material. Aldehyde and/or Osmium vapor fixation would likely be less disruptive.
My FD time/temp schedule was: 48hrs-at--80C 36hrs-at--60C 24hrs-at--40C 12hrs-at--30C 12hrs-at--15C 24hrs-at--5C 6hrs-at-+10C 30hrs-at-+20C Vacuum was maintained with a cryo sorption pump.
I can post some images to my web page by the end of the week, if you are interested. The unfixed yeast pics are there already. Follow the links from Research Projects to Sample prep for SEM... http://www.personal.psu.edu/ejb11
cheers ed
Edward J. Basgall, PhD The Pennsylvania State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618 http://www.personal.psu.edu/ejb11/
Let me modify my earlier suggestion for using the PrintScreen copying method to concur with Rainer's suggestion below. The PrintScreen trick still works, but other methods are probably more suitable when available. And this package does allow for copying the spectrum to the Windows clipboard.
You should be advised that you will probably have to use the Paste Special function to select the spectrum "picture" for pasting instead of the sample id "text". The text will come up as default.
At 08:51 PM 9/26/97 +0200, Rainer wrote: } } } Malgorzata Warmuzek wrote: } } } I have a problem with an import the file with an extension *.sp ( spectrum ) } and put it to the } } document in word 6 or amipro } } Dear Malgorzata, } } I use the ISIS suite 3.0 and there is an easy way to copy spectra to any other } windows program, maybe it works also with your version. Use the command } 'Buttons' 'Print' and a new window will appear called 'Spectrum printing'. } There it is possible to activate the command 'Edit' 'Copy'. The Spectrum will } now be transfered to the clipboard in form of a vector graphic. } } Kind regards } } Rainer ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering, or 270 Metals Development Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-3091
Dear Colleagues I'm trying to cut 60 um parafin sections, but I've found it very difficult. It's rare to botain a good section There is some procedure to facilitate this work
Francisco Javier Hernandez Blazquez Centre commun de Quantimetrie, 8 avenue Rockefeller 69373 LYON CEDEX 08. France. Tel : (33) 4 78 77 75 19.
Responding to the message of {v03007804b05554e7eb94-at-[206.69.208.21]} from Yves Maniette {yves-at-giga.sct.ub.es} (by way of Nestor J. Zaluzec): } Following the thread about coating of a glass sample, Jacky Larnould added } off line that when the objective aperture is set (above the sample) the } "charging effect" disappears, and the sample does no break. While I have } noticed this phenomenon for a while I have never come with a satisfactory } explanation. Has anyone found a good explanation to this? } } Yves Maniette } We looked at this some time ago. JEOL machines work better than Philips, unless you install a really large aperture (we use 800 micron) and move to that aperture rather than moving the apertures out of the beam. The lack of symmetry induced by moving the aperture rod to the side rather than moving to an empty hole prevents the system from effectively stabilizing the charge, and the beam is displaced from the specimen.
The beneficial effect of the aperture is probably due to some capacitative charge dissipation from the sample by the very close proximity of the aperture. It is important to have everything properly aligned - beam and aperture, or the lack of radial symmetry will again deflect the beam away from the area of interest.
Different specimens respond differently - we have had very good success with single crystal alumina, but less success with polycrystalline specimens.
Good luck.
Stuart
__________________ Stuart McKernan stuartm-at-tc.umn.edu Microscopy Specialist CIE Characterization Facility, University of Minnesota Phone: (612) 626-7594 100 Union Street S. E., Minneapolis, MN 55455 Lab: (612) 624-6590
We are trying to calculate the surface area of gap junctions. Has anyone done something similar to this? Are the gap junctional complexes (groups of connexons) arranged in a circular profile? If you could provide me with a reference or any information, I would appreciate it.
With regard to the following: ================================================= We are looking for any information we can find on the } microscopy/identification of particles from automobile tires. Seems we } have a case where we need to try to match particles from a pair of tennis shoes to a particular set of automobile tires(best case), or at least identify particles as possibly coming from automobile tires (more likely). ================================================= Assuming what you are saying is you have a need to demonstrate "common origin", which is not all that of an uncommon request, we have found the best approach is to use thin section TEM on the particulates. Tire-origin particulates have a nice characteristic dispersion of carbon black plus other inorganics.
The elastomeric system found in tennis shoes also has inorganic additions present. Generally speaking (based on our own in-house experience) the additive particles are larger than would be the carbon black and other particles found in a tire. Hence to differentiate between tires vs. tennis shoes, this should not be a major problem. If you wanted to show that the particulates came from a specific tire or a specific pair of tennis shoes, this would be a much higher level question, and considerable additional work (and the running of far more samples) would be indicaed.
The reason why this is more of a TEM than SEM request is that the particulates, especially the carbon black, is really below the practical limit of SEMs in terms of resolution and furthermore, the TEM view of this kind of sample is far easier to understand and interpret. And when EDS does have to be done, that data also is far easier to interpret and understand (because you don't have to deal with "depth" effects).
All thin sectioning on these kinds of samples must be done with the use of a good ultramicrotome, using a cryo stage, and using diamond knives, and we would always use a "Materials Science" diamond knife in order to not unnecessarily balloon up the cost of doing this kind of work.
Disclaimer: Our Structure Probe laboratories offer this kind of laboratory analytical service for clients needing to have done this kind of work. We are also a major provider of "materials science" diamond knives for persons wanting to do this kind thin sectioning.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Charlie Kong wrote: } } Rainer, } } Have you found that the spectrum you got it by copying become a low resolution (287x287 pixels) graph? } } I have tried the method you described in the internet, and felt the difficulty to see the lebels of axis... } } p.s. I pasted the spectrum in Paint Shop Pro. } } Charlie Kong kong-at-t-rex.materials.unsw.edu.au
Dear Charlie,
The spectrum is copied to the clipboard in form of a vector graphic. PaintShopPro is a pixel graphic program. So the spectrum is converted when it is imported. You can import the graphic directly to your Word processor. In the german version of Winword it is done by the command 'Einfugen' 'Inhalte Einfugen' 'Graphic'. Afterwards it is possible to double click the graphic and to change e.g. labels.
Kind regards
Rainer
------------------------------------------- Rainer Ziel Akzo Nobel Central Reasearch 63784 Obernburg - Germany
On Mon, 29 Sep 1997, Francisco Javier Hernandez Blazquez wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear Colleagues } I'm trying to cut 60 um parafin sections, but I've found it very difficult. } It's rare to botain a good section } There is some procedure to facilitate this work
A neat old trick for doing this is to use rubber cement in the mixture. It gives the paraffin the needed tensile strength to prevent crumbling.
David Feldser asked for references on microwave-accelerated staining for electron microscopy. I list 14 references FYI:
1. Estrada JC, Brinn NT, Bossen EH: A rapid method of staining ultrathin sections for surgical pathology TEM with the use of the microwave oven {italic} . {/italic} Am J Clin Pathol 1985, 83:639-641
2. Zondervan PE, de Jong A, Sorber CWJ, Kok LP, de Bruijn WC, van der Kwast TH: Microwave stimulated incubation in immunoelectron microscopy: a qualitative study {italic} . {/italic} Histochem J 1988, 20:359-364
3. Matsutani S, Yamamoto N: Improved methods for immuno-electron microscopy of cultured cells: use of a novel substrate and application of microwave irradiation {italic} . {/italic} Acta Histochem Cytochem 1990, 23:227-236
4. Wouterlood FG, Boon ME, Kok LP: Immunocytochemistry on free-floating sections of rat brain using microwave irradiation during the incubation in the primary antiserum: light and electron microscopy {italic} . {/italic} J Neurosci Methods 1990, 35:133-145
6. van Deuren B, van Reempts J, Borgers M: Microwave-enhanced silver staining of degenerating neuronal processes {italic} . {/italic} Eur J Morphol 1992, 24:597(abstract)
7. Giammara BL, Hopfer RL, Yates PE, Hanker JS: A rapid silver stain for the DNA of microorganisms cultured from AIDS or other immunocompromised patients {italic} . {/italic} Proc Twelfth Int'l Congr Electron Micros 1990, 48:762-763
8. Hanker J, Giammara B: Microwave-accelerated cytochemical stains for the image analysis and the electron microscopic examination of light microscopy diagnostic slides {italic} . {/italic} Scanning 1993, 15:67-80
11. Utsunomiya H, Shan L, Kawano I, Iwasaki A, Ono K, Kobayashi A, Kuma K, Kishikawa S, Kakudo K: Immunolocalization of parathyroid hormone in human parathyroid glands with special references to microwave antigen retrieval {italic} . {/italic} Endocr Pathol 1995, 6:223-227
12. Stirling JW, Graff PS: Antigen unmasking for immunoelectron microscopy: labeling is improved by treating with sodium ethoxide or sodium metaperiodate, then heating on retrieval medium {italic} . {/italic} J Histochem Cytochem 1995, 43:115-123.
13. Login GR, Dvorak AM. Microwave fixation and microwave staining methods for microscopy. In: Hayat MA ed. Immunogold-silver staining: methods and applications. Boca Raton, CRC Press, 1995, pp. 163-182
14. Login GR, Dvorak AM: The Microwave Toolbook. A Practical Guide for Microscopists. Boston, Beth Israel Hospital, 1994, pp. 184
Reminder: "Optimizing Light Microscopy" will be hosted by the Biology Department of Providence College this Friday, October 3. Course cost: $150,includes a copy of "Optimizing Light Microscopy for Biological and Clinical Laboratories". Despite the title of the book, the course has wide application to light microscopy in any venue.
For further information, write, call, or email:
Barbara Foster President Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108 PH: (413)746-6931 FX: (413)746-9311 email:mme-at-map.com --------------------------------------------------------------------------------------------------------------------------------- ********** Microscopy/Microscopy Education ********** America’s First National Consortium of Microscopy Experts Specializing in Customized, On-site Training in all areas of Microscopy, Sample Prep, and Image Analysis
Reminder: "Optimizing Light Microscopy" will be hosted by the Biology Department of Providence College this Friday, October 3. Course cost: $150,includes a copy of "Optimizing Light Microscopy for Biological and Clinical Laboratories". Despite the title of the book, the course has wide application to light microscopy in any venue.
For further information, write, call, or email:
Barbara Foster President Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108 PH: (413)746-6931 FX: (413)746-9311 email:mme-at-map.com --------------------------------------------------------------------------------------------------------------------------------- ********** Microscopy/Microscopy Education ********** America’s First National Consortium of Microscopy Experts Specializing in Customized, On-site Training in all areas of Microscopy, Sample Prep, and Image Analysis
Reminder: "Inside Fluorescence", a lecture-demonstration on factors affecting fluorescence microscopy, will be hosted this SATURDAY, OCTOBER 4, by Providence College.
Cost of class: $150 which includes a detailed course workbook.
For further information, please contact our office.
Barbara Foster President Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108 PH: (413)746-6931 FX: (413)746-9311 email:mme-at-map.com --------------------------------------------------------------------------------------------------------------------------------- ********** Microscopy/Microscopy Education ********** America’s First National Consortium of Microscopy Experts Specializing in Customized, On-site Training in all areas of Microscopy, Sample Prep, and Image Analysis
The best explanation for the effect of the objective aperture was published (I recall) in a Philips bulletin.
The section in a TEM has a positive charge generated in it as the primary beam produces secondary electrons in the section which then exit from it. This charge will destroy the section if it is not neutralised or conducted away through e.g. a carbon coat.
The objective aperture neutralises the positive charge in the specimen by reflecting back backscattered and secondary electrons which re-enter the section.
Mel Dickson Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
Dear Yves, In every TEM I've seen, the objective aperture slides in just below the sample. In samples that are likely to break, such as formvar-covered slots, I was told never to look at the sample without the objective aperture in. Having all the high-kV electrons hitting on just the top side would pop the film. If the objective aperture is in, the high-kV electrons scattered back from the aperture below the specimen will balance the flux from above and the film probably won't break. BTW, it doesn' matter which side of the sample you coat, since the electrons go right through, anyway. Otherwise, it wouldn't be TEM. You wrote: } All, } } Following the thread about coating of a glass sample, Jacky Larnould added } off line that when the objective aperture is set (above the sample) the } "charging effect" disappears, and the sample does no break. While I have } noticed this phenomenon for a while I have never come with a satisfactory } explanation. Has anyone found a good explanation to this? } } Yves Maniette } Regards, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am interested in purchasing some time on a commercial ESEM to do some microscopy of fractures. Does anyone know of any ESEMs in CA, AZ, OR or WA that will provide commercial time? If so please email me at HWachob-at-fail.com. Thank You for your assistance.
Problem: I do have a excellent Zeiss stand and a complete series of Leitz planapochromatic objectives. However as you probable know Zeiss microscopes have a 160 mm mechanical tubelength whereas teh Leitz objectives are calculated for 170 mm. What kind of correction lens can I use in between the tube so that I can use the Leitz lenses on the Zeiss stand without loose of optical quality? Can anybody give me a good advice .
feldsdm4-at-juniata.edu wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Email: feldsdm4-at-juniata.edu } Name: David Feldser } } School: Juniata College } } Question: Where can i find a good source of information } about microwave staining technology for the } electron mircroscope? } } ---------------------------------------------------------------------------
Hi David,
this Link might be helpful http://www.rfglobalnet.com
--- ImmunoFluorescence Controls for Renal Biopsies------ From T. Fores: } Anyone doing immunofluorescenst techniques in renal biopsies running a control? } } From: Wanda } Hi Teresa: } } At the present we are not running a control on our immunofluorescent } techniques in renal biopsies. Has anyone replying back doing this? Even if we pooled resources, there would not be enough material to distribute to all laboratories that run this test. One solution is to run a known positive patient sample at the time titre the antibody. Another suggested solution was to run a tonsil control with each batch of patient samples (simplifying solution - the test material in each tissue is equivalent). And what about the control for the complement (C1, C3, C4) and fibrinogen?
Business idea: rats with immune mediated GN, frozen kidney samples on slides ...
The situation regarding immunofluorescence controls (IF) for renal biopsies is an example of the tension that exists between pure science and the practical application of science in clinical laboratories. Manuals are written and regulations are mandated without regard to the practical aspect of running a test in a clinical laboratory. Some rational solution is needed to stop laboratorians from resorting to draconian measures in order to comply with the regulatory agencies.
} Also would you know of anyone who might be interested in serving as a } consultant for EM or an EM Tech interested in relocating? I can consult on Transmission EM of human tissues. |--------------------------------| | Karlene Hewan-Lowe, M.B., B.S. | | Department of Pathology | | Emory University | | Phone: 404-686-2926 | | Fax: 404-6864978 | |--------------------------------|
Message text written by INTERNET:MWIS-at-crf.cuis.edu } microscopy-at-Sparc5.Microscopy.Com {
Check out PAX-it, an excellent image capture, archiving and databasing system. It has pretty powerful report generation software which allows y= ou to effectively replace traditioinal film in the photo documentation process. Going digital seems to pay for itself quickly. The other good thing is that it has network software, so you can access the database of images from your desktop PC's.
------------------------------------------------------------------------ If you work in MATERIALS R&D, SOLID STATE PHYSICS, CRYSTALLOGRAPHY or in a SCIENTIFIC LIBRARY, this is interesting for you.
A true compilation of all world literature data on MATERIALS CONSTITUTION; PHASE DIAGRAMS and RELATED DATA is available in print: the annual "RED BOOK" series . It started with the publication year 1990 and is produced by MSI and the Russian information service VINITI.
This compilation extracts data, diagrams and text from the world literature and arranges the information after alloy systems (not after publications), every year. - No need to browse hundreds of journals + proccedings - All foreign language publications translated into English - Best coverage of "eastern" publications available anywhere. - Uniformly structured detailed summaries of binary, ternary,..., multicomponent systems.
The introduction package, comprises the publication years 1990 to 1995 inclusive. It provides over 5300 summaries, more than 6000 diagrams and more than 1200 tables printed on over 8500 pages. Have a look at sample summaries and order information at
http://www.msiwp.com or inquire at info-at-msiwp.com
NOTE: The very attractive introduction offer expires mid December `97.
------------------------------------------------------------------------- This mailing list will inform you on the global phase diagram evaluation program of MSI and its team MSIT. It will announce how you can access its further products, such as the literature data base, electronic phase diagrams, etc. The traffic will be as low as 1 or 2 mailings per quarter. If this information is of no interest to you, please accept our appologies for the present message and send an e-mail to majordomo-at-msiwp.com with the message in the body: unsubscribe msi-adverts This will remove your address from the list.
Dear Microscopists I'm just beginning to use LR white as embedding medium for TEM- immuno- cytochemistry. For previous embeddings of plant root segments for conventional TEM research with SPURRs resin, I used flat embedding mold of silicon rubber for simple orientation of the segments. Is it possible to use flat embedding mold also with LR white and how is it possible to avoid contact with oxygen during polymerization? What kind of molds are otherwise most suitable for LR white? Furthermore I'm looking for a common embedding procedure of plant roots in LR white for use in TEM immunogold-labelling. I would like to contact in this way a lab with similar interests. Thanks to all comments.
Martin Bartels FB Biologie / AG Pflanzenoekologie C.v.O. University Oldenburg /Germany PO Box 2503 26111 Oldenburg phone ++49 441 798 3436 fax ++49 441 798 3436 e mail: bartels-at-uni-oldenburg.de
Here is a copy of replies I got about teh charging effect. One was sent off line. I made an horrible mistake saying that the aperture is above the sample, which is nonsense. Well let's say that I had a cold yesterday. YM.
All,
Following the thread about coating of a glass sample, Jacky Larnould added off line that when the objective aperture is set (below the sample) the "charging effect" disappears, and the sample does not break. While I have noticed this phenomenon for a while I have never come with a satisfactory explanation. Has anyone found a good explanation to this?
capacitance effect between the sample and aperture - the smaller the aperture the better for diffraction burgers vector analysis. Move the aperture around and note what happens. Also compare Philips CM and Jeol aperture geometries - you should find Philips easier to use with non conducting samples owing to aperture being nearer to specimen. It's been a while since I was doing this, however, when setting up diffraction conditions, it's best to use large objective aperture when locating yourself in the pattern. Removing the aperture entirely, as normally done, causes massive beam tilt in strongly charged samples (e.g. dirty MgO). cheers
We looked at this some time ago. JEOL machines work better than Philips, unless you install a really large aperture (we use 800 micron) and move to that aperture rather than moving the apertures out of the beam. The lack of symmetry induced by moving the aperture rod to the side rather than moving to an empty hole prevents the system from effectively stabilizing the charge, and the beam is displaced from the specimen.
The beneficial effect of the aperture is probably due to some capacitative charge dissipation from the sample by the very close proximity of the aperture. It is important to have everything properly aligned - beam and aperture, or the lack of radial symmetry will again deflect the beam away from the area of interest.
Different specimens respond differently - we have had very good success with single crystal alumina, but less success with polycrystalline specimens.
In every TEM I've seen, the objective aperture slides in just below the sample. In samples that are likely to break, such as formvar-covered slots, I was told never to look at the sample without the objective aperture in. Having all the high-kV electrons hitting on just the top side would pop the film. If the objective aperture is in, the high-kV electrons scattered back from the aperture below the specimen will balance the flux from above and the film probably won't break. BTW, it doesn' matter which side of the sample you coat, since the electrons go right through, anyway. Otherwise, it wouldn't be TEM.
The best explanation for the effect of the objective aperture was published (I recall) in a Philips bulletin.
The section in a TEM has a positive charge generated in it as the primary beam produces secondary electrons in the section which then exit from it. This charge will destroy the section if it is not neutralised or conducted away through e.g. a carbon coat.
The objective aperture neutralises the positive charge in the specimen by reflecting back backscattered and secondary electrons which re-enter the section.
To: Manufacturers of microscopes, imaging and image analysis systems, and sample preparation equipment
ASCB has informed us that there is still space available for MME to conduct market research at the upcoming Cell Biology meeting. If there is enough interest from you, we would be pleased to conduct a multi-client survey, by subscription. This meeting provides a prime venue for testing new ideas for instrument development. Since MME conducted research at this meeting in 1993, there are also select opportunities for long-term trend evaluation.
October 9th will be the deadline for our decision to go. Please note that there will be only one block of questions available per technology area and a total of approximately 25 questions, so space will be limited to first come/first served.
For further information, please respond directly to:
Barbara Foster Consortium President Microscopy/Marketing & Education 53 Eton Street Springfield, MA 01108 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com
Message text written by Randy Schnack } Microscopy-at-Sparc5.Microscopy.Com {
The Leco system is the one that most are using. It uses a high res CCD camera to provide film replacement using digital means. The software the= y provide is PAX-it, which does the archiving in an easy cabinet & folder style database. It has a really cool "report generation" tool that links=
all of the data fields with the images in MS Word and gives you a great print-out. =
The budget of 15k sounds about right for a complete computer, capture car= d, software AND high res camera. If the user wants to provide their own pentium, that will save some money as well.
Dear Yves, } } One side carbon coating is sufficient. It can be on the top or the down } side in the microscope, though you should get a better image if you place } it on top,
The image should be the same regardless of which side is coated. Assuming a perfect plane-wave for the incident beam, coating the bottom will cause the image--assumed to be perfect--to be convoluted with the scattering from the carbon; whereas, coating the top will form the image with a beam which has been convoluted with the same scattering. Both resulting images will be the same. Yours, Bill Tivol
Greetings to all, is there any person or EM-Lab out there who/which uses/used a resin = formulation with EPON 815 (Polysciences).=20 Would be glad to receive a mixing recipe which works/worked as I have a = small amount left for testing a staining procedure on semithin sections = made from original Epon embedded blocks. If also a reference could be = added it would be fine. Thanking you in advance Wolfgang MUSS, EM-Lab, Dept. Pathology, A-5020 SALZBURG/AUSTRIA, Europe e-mail: W.Muss-at-lkasbg.gv.at
This is a question for the Noran Voyager 4 users. After I obtain an image of my sample, on the monitor, is it possible to e-mail it from the Voyager to a person on MS Exchange? Does it have to be converted into a particular type of file? Does the person on the other end, the one receiving the image, have to convert it and then view as a certain type of file? Are instructions to do this in the Voyager manuals?
Hi would it be simpler to put a correction collar on the scope that would extend your tube length to 170mm?
Bob
On Mon, 29 Sep 1997, P.M. HOUPT wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } dear microscopists, } } Problem: I do have a excellent Zeiss stand and a complete series of } Leitz planapochromatic objectives. } However as you probable know Zeiss microscopes have a 160 mm mechanical } tubelength whereas teh Leitz objectives are calculated for 170 mm. } What kind of correction lens can I use in between the tube so that I can } use the Leitz lenses on the Zeiss stand without loose of optical } quality? } Can anybody give me a good advice . } } thank you in advance, } } Pieter Houpt } } (the Hague ,the Netherlands) } } }
I flat embed LR White all the time. I got an old Vacume oven out of surplus and hooked up a vacume pump and a dry nitrogen tank. When it is time for polymerization and the temperature is stable at 55C, I purge the chamber 3 times with nitrogen by pumping it down and letting in the nitrogen. On the last perge, I fill the chamber with the nitrogen leaving a slight vacume 1-3 lbs. Just to keep the door sealed. Polymerize for 24-48 hrs.
I use the peel-away polypropylene embedding molds. Dont use polystyrine.
On Wed, 1 Oct 1997, Martin Bartels wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear Microscopists } I'm just beginning to use LR white as embedding medium for TEM- immuno- } cytochemistry. For previous embeddings of plant root segments for } conventional } TEM research with SPURRs resin, I used flat embedding mold of silicon } rubber for } simple orientation of the segments. Is it possible to use flat embedding } mold } also with LR white and how is it possible to avoid contact with oxygen } during } polymerization? What kind of molds are otherwise most suitable for LR } white? } Furthermore I'm looking for a common embedding procedure of plant roots in } LR } white for use in TEM immunogold-labelling. I would like to contact in this } way } a lab with similar interests. } Thanks to all comments. } } Martin Bartels } FB Biologie / AG Pflanzenoekologie } C.v.O. University Oldenburg /Germany } PO Box 2503 } 26111 Oldenburg } phone ++49 441 798 3436 fax ++49 441 798 3436 } e mail: bartels-at-uni-oldenburg.de } }
There is a paper from 1987 describing how to make a simple glow-discharge unit. We made one from this design and have been using it successfully for years, slightly modified - the needle-type valve doesn't seem to be necessary, just close off the feed for the argon with a removable clamp. The reference is : Aebi U. and Pollard T.D., A glow discharge unit to render electron microscope grids and other surfaces hydrophilic. J. Electron Microsc.Technique, 7:29-33 (1987). Hope this helps. Lesley Weston.
On Thu, 25 Sep 1997, Gunnel Karlsson wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello, } } I don't have access to a nice workshop, where they can construct a glow } discharger, where in Europe can I buy one? Or, is it out there someone, who } has an old mashine you don't need anymore? } } TIA } } Gunnel Karlsson } } ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ } Gunnel Karlsson E-mail Gunnel.Karlsson-at-oorg2.lth.se } Biomicroscopy Unit Tel +46 222 8229 } Inorganic Chemistry 2 Fax +46 222 4012 } Box 124 } S-221 00 LUND, Sweden } ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ } This message was sent by Eudora with recycled electrons } } }
There is a paper from 1987 describing how to make a simple glow-discharge unit. We made one from this design and have been using it successfully for years, slightly modified - the needle-type valve doesn't seem to be necessary, just close off the feed for the argon with a removable clamp. The reference is : Aebi U. and Pollard T.D., A glow discharge unit to render electron microscope grids and other surfaces hydrophilic. J. Electron Microsc.Technique, 7:29-33 (1987). Hope this helps. Lesley Weston.
On Thu, 25 Sep 1997, Gunnel Karlsson wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello, } } I don't have access to a nice workshop, where they can construct a glow } discharger, where in Europe can I buy one? Or, is it out there someone, who } has an old mashine you don't need anymore? } } TIA } } Gunnel Karlsson } } ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ } Gunnel Karlsson E-mail Gunnel.Karlsson-at-oorg2.lth.se } Biomicroscopy Unit Tel +46 222 8229 } Inorganic Chemistry 2 Fax +46 222 4012 } Box 124 } S-221 00 LUND, Sweden } ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ } This message was sent by Eudora with recycled electrons } } }
At 09:17 AM 9/30/97 -0400, James Martin wrote: } Can anyone steer me to a website that has photomicrographs of histological } samples; in particular, blood samples?
There are a number of such sites. There are annotated links to the best of them from "The Histotech's Home Page" (http://www.histology.to). Go to "Peggy's Links" and select the section on images.
Best regards, Steven Slap ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
Some years ago our lab studied substance P and did some immuno-EM. There is a nicely detailed experimental procedure in this paper: "A substanceP-like peptide in bullfrog autonomic nerve terminals: anatomy biochemistry and physiology," C.W.Bowers, L.Y. Jan & Y.N. Jan, Neuroscience, vol 19, No 1, pp343-356, 1986. Good luck! Larry D. Ackerman (415) 476-8751 Howard Hughes Medical Institute FAX (415) 476-5774 UCSF, Box 0724, Rm U426 533 Parnassus Ave. mishot-at-itsa.ucsf.edu San Francisco, CA 94143
O.k., with all the positive comments about the Epson Stylus Photo printer, I have one more question: Since the postscript level 2 emmulation for the Stylus photo adds ~ 20% to the cost of the printer is it worth it? Every other printer I have has postscript capabilities and we generally use it (even though we're Intel PC / Windows based, not Macintosh) any opinions would be great! Thanks.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"640K ought to be enough for anybody." -- Bill Gates, 1981
This WWW site has hundreds of histology images, including several related to blood samples.
Best Regards,
Kirk J. Czymmek University of Delaware kirk-at-udel.edu
On Tue, 30 Sep 1997, James Martin wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Can anyone steer me to a website that has photomicrographs of histological } samples; in particular, blood samples? } } Thanks. } } James Martin } Williamstown Art Conservation Center } } }
Martin Bartels wrote: ================================================ I'm just beginning to use LR white as embedding medium for TEM- immuno- cytochemistry. For previous embeddings of plant root segments for conventional TEM research with SPURRs resin, I used flat embedding mold of silicon rubber for simple orientation of the segments. Is it possible to use flat embedding mold also with LR white and how is it possible to avoid contact with oxygen during polymerization? What kind of molds are otherwise most suitable for LR white? ================================================== One can use flat UV transparent silicone molds for this type of work. However, since the (transparent) silcone, in order to be UV transparent, does not have any of the additives normally incorporated to provide chemical resistance, don't expect long lifetimes, some people report being able to use a cavity (with L. R. White(TM) for example) only once or twice. The problem with oxygen exposure is easily "solved" by over filling the cavity with resin slightly, so there is a positive miniscus, and then placing another identical transparent mold on top, flat side down onto the over- filled cavities. Capillary action ensures that there is a quite adequate seal against the presence of oxygen.
Such molds can be purchased at the main suppliers of accessories of consumables for microscopy laboraotries including SPI, full details about which can be found on the SPI website, given below. These molds do not all "come out of the same source" and are not all the same, so don't assume the result experienced from one brand would be the same for all other brands.
Disclaimer: SPI Supplies manufactures transparent silicone molds for this application and we would obviously have an interest in seeing more people using these transparent molds.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
"Electron, X-ray and Ion Spectroscopies-A Primer", an all day educational seminar, will be presented Friday, November 14, 1997, at Argonne National Laboratory. Advance registration is required. For more information contact Chas. Allen at allen-at-aaem.amc.anl.gov or Anita Brandes at g10809-at-email.mot.com including your complete mailing address. We will send you information and a registration form. Registration fee is $30 ($10 for students) which includes lunch and refreshments at breaks.
======================================== Charles W. Allen Electron Microscopy Center-HVEM-Tandem Facility MSD 212/E211 Argonne National Laboratory Argonne. IL 60439 USA
Email:allen-at-aaem.amc.anl.gov Tel: 630-252-4157 Fax:630-252-4798 (Note: On August 3,1996, Area Code changed from 708 to 630) ========================================
I'm trying to find a protocol that describes the TOTO or PATOTO (not potato!) technique for fixing plant specimens for SEM. I've tried doing a literature search and all I get are references to Dorothy and Oz, just kidding. I've read about the procedure but I cannot find a good descriptive protocol. Thanks for helping with this.
I'll get that pretty picture, and it's little dog, too.
Hoping a house doesn't fall on you,
Paula = )
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
To everyone with an e-mail address in my computer (sorry if you have this information already):
ALWYN EADES
(full name: John Alwyn Eades)
IS MOVING TO LEHIGH UNIVERSITY IN OCTOBER 1997 (moving in about ten days)
Present address
Materials Research Laboratory 104 S Goodwin Urbana Illinois 61801-2985
is moving to
Department of Materials Science and Engineering Lehigh University Whitaker Laboratory 5 East Packer Avenue Bethlehem PA 18015-3195
610 758 4231 610 758 4244 FAX jae5-at-lehigh.edu not activated yet. ** Alwyn Eades Center for Microanalysis of Materials University of Illinois at Urbana-Champaign Phone 217 333 8396 Fax 217 244 2278 eades-at-uimrl7.mrl.uiuc.edu (NB those are letter l not ones)
In mid-October this year (1997), I will be moving to Lehigh. The new address will be:
Department of Materials Science and Engineering Lehigh University Whitaker Laboratory 5 East Packer Avenue Bethlehem, PA 18015-3195
Peter: There is a more serious problem which I am sure the Zeiss and Leitz people will quickly point out. That is that the older series of objectives had to be used with a corresponding ocular to reduce optical aberrations, etc. in the objectives, giving you that high optical quality. Really old scopes had a way of adjusting the tube length, but that is the least of your problems in a more modern scope when you switch to another manufacturer's objectives. Of course the latest scopes have "infinity corrected" optics, but I wouldn't switch them either. If you use them, you will not have as good an image, but it would give you an image. The best tack would be to find an appropriate Leitz stand with appropriate oculars. Many older scopes are available, and without the optics are pretty cheap.
Regards, Mike
} } dear microscopists, } } } } Problem: I do have a excellent Zeiss stand and a complete series of } } Leitz planapochromatic objectives. } } However as you probable know Zeiss microscopes have a 160 mm mechanical } } tubelength whereas teh Leitz objectives are calculated for 170 mm. } } What kind of correction lens can I use in between the tube so that I can } } use the Leitz lenses on the Zeiss stand without loose of optical } } quality? } } Can anybody give me a good advice . } } } } thank you in advance, } } } } Pieter Houpt } } } } (the Hague ,the Netherlands) ================================================ Michael L. Boucher Sr. mboucher-at-isd.net 13345 Foliage Avenue Apple Valley, MN 55124-5603 Ph 612-432-8836 ================================================
David Feldser wrote: ================================================= Question: Where can i find a good source of information about microwave staining technology for the electron mircroscope? ================================================= The most often asked for book in this category, at SPI, is the following:
The Microwave Tool Book Authors: G. R. Login, DMD and A. M. Dvorak, MD http://www.cccbi.chester.pa.us/spi/catalog/books/book29.html
You can see the entire Table of Contents as well as some comments by reviewers, just to make sure this is what you really want.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Hello, I am an undergrad attending the University of Scranton. I am trying to find some beginner information regarding flourescent microscopy. While I have found some web-sites dealing with flourescence, they are all too advanced for me. It is the same situation with the school's library. Most of the information is not geared for the beginner. I am mostly interested in staining neural tissue (CNS).
Thank you very much for any help you can offer. It will be greatly appreciated.
Several list members have requested a summary of the responses to my inquiry after websites with histological images. Thanks again to all who responded, and apologies if I've inadvertently not included your response in this summary. Here goes:
- snip
The following sites contain histological images (including blood):
Scott Henderson, Ph.D. Director of Microscopy, Mount Sinai School of Medicine, Department of Cell Biology & Anatomy
- snip
There are a number of such sites. There are annotated links to the best of them from "The Histotech's Home Page" (http://www.histology.to). Go to "Peggy's Links" and select the section on images.
This WWW site has hundreds of histology images, including several related to blood samples.
Best Regards,
Kirk J. Czymmek University of Delaware kirk-at-udel.edu
- snip
try at http://www.cba.arizona.edu/histology-lab.html
or http://www.hslib.washington.edu/courses/blood/
or at http://144.92.79.188/histology/histo.html
good luck
Gabriel Adriano Rosa Area Microscopia Electronica, Depto. Cs. Biologicas Fac. de Ciencias Exactas y Naturales, Universidad de Buenos Aires Ciudad Universitaria, 4 piso, Pab. II, CP 1428, Buenos Aires, ARGENTINA
- snip
My Histology web page has a collection of links - they ought to get you what you want or at least send you in the correct direction.
} Date: Wed, 01 Oct 1997 11:06:53 -0400 } From: Edgar Voelkl {vog-at-ornl.gov} } X-Sender: vog-at-cosmail1.ctd.ornl.gov } To: "Dr. Larry F. Allard" {allardlfjr-at-ornl.gov} } MIME-version: 1.0 } Status: O } } Dear microscopists, } } I regret to inform you that Professor Gottfried Moellenstedt passed away } on September } 11th, close to midnight. As the father of the electron biprism he led } developments in the area of electron holography -- as well as } many other areas ! -- and contributed strongly to the reputation of the } University of Tuebingen, Germany, in the international community. } Professor } Moellenstedt will be missed by his many students and long-time } collaborators and colleagues. } } } } Dr. Edgar Voelkl } ORNL } Bldg 4515 } 1 Bethel Valley Road } P.O. Box 2008 } Oak Ridge, TN 37831-6064 } } Tel.: (423) 574-8181 } Fax: (423) 574-4913 } email: vog-at-ornl.gov }
Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
Hello to all, especially dear Rosey (AUSTRALIA) and Dr. GARBER (SPI, = USA), thank you two for your suggestions and greetings (espec. Rosey, for your = hello to the "lippizaners" (for all those not knowing what they are: = those being totally white colored, only male horses, serving for the = famous "Spanische Hofreitschule"- i.e. "Spanish k.k. royal riding = school" - in VIENNA/Austria since about 200 years now) but for all:answering in short for elucidation of my problem: 1) (for Dr. GARBER/SPI:) I meant to say: EPON 81 5 (eight fifteen); it = is a remnant of resin given to me without costs from Polysciences at the = trinational EM-congress at REGENSBURG (FRG, in Sept. 1997) 2) Rosey: thank you very much for sending me a recipe of your mixture = (find below my mixture) and the LUFT-reference.: do you use any left = lot(s) of originally EPON 812 or a substitute of this? If this is true, = would you=20 (or anyone, who uses or has in stock remainders of the original MONSANTO = / SHELL resin component) be so kind to send at a maximum of about 100 g = of that "old", original Epon 812 by gratitude to my Lab (adress given = below)???? 3) The problem, generally adressed, is: The production of the original resin component EPON 812 (Trade mark = of MONSANTO/SHELL) has been cancelled. No one -as to my knowledge- can order EPON 812 any = more. The substance EPON 812 has been replaced by chemically very = similar components (like SERVA/Bioproducts, Boehringer/Ingelheim: = Glycidether 100, which I am using now). A lot of Trade names have been = established by several supplying companies: EMBED 812, POLARBED 812, = SPI-Pon 812 ("exact direct replacement"). I have developed for use in our Lab about 10 years ago a simple, = polychromatic two-step staining for semithin sections, which was devised = for our needs with original EPON 812 resin embeddings. Since 1995 the = intended and standardized staining procedure failed to produce same = results in staining intensity like before. A long time I tried to = overcome that failing in looking forward to "unusual specimen = processing" (which was standardized at that time, too). Nothing happened = until I remembered as the only alteration in our standardized processing = the changing of resin component ORIGINAL Epon 812 (which I had in stock = for about 2 years more than being produced or available from suppliers) = to the replacement by "glycid ether 100 (SERVA)", which is, by = certificate, a 1,2,3-Propanetriol glycidyl ether with a MW of ~ 306.0 = and an epoxy equivalent (g/mol) of 150. Viscosity is certified as 196 = mPa.s at 25 degr. centigrade. I had to change and newly standardize the staining procedure in a very = hard labor, now including 2 short steps more, to get same results as = before. As an explanation for that I asssume the polymerizing character = of the new, replaced substance to be altered, leading to altered = staining properties of the sections too. This interpretation at the = moment is the only, because I had seen and noted an unusual changing of = resin color when mixing up the first three components (intensive red = color!; see below), which fades only to the (with old EPON 812) = "formerly seen yellowbrown color unless mixing components at least for, = say, 20 min, oxygen access provided (if overlayed by freon gas, to = shield from an increased humidity, this will last at least ~ 60 min!). = This was the only difference in my processing I could find! But there is = no difference in using a "red" or "yellowbrown" resin mixture (let me = assume it to be "unoxidized"/"oxidized" resin components): The routine = staining never achieved the same results without adding the two steps = "more". As I am going to prepare a manuscript for a paper on subject = "polychromatic staining of semithin epoxide ("EPON") sections", I should = be prepared to answer the question: "why the staining procedure formerly = did it without, and why now you need unalterable 2 steps more". So I am = trying to lock my hypothesis on altered polymerization properties of my = resin in staining "old fashioned EPON 812"-semithin sections, as = compared to variable EPON replacements. 3a) Hope, that you, Dr. Garber could supply me with any small amounts=20 (up to, say maximum 50 grams) of your SPI -Epon Replacement resin(s) = (kits?) to be able to perform my tests.
4) Rosey: My EPON-(Glycidether 100) mixture=20 ( based on the formerly used EPON 812, WPE ~ 146-150, according to the = A, B-formulations of LUFT 1961 ) is (measurement by weight):
45,23 g Glycidether 100 (SERVA =3D Bioproducts, Boehringer/Ingelheim, = FRG) + 13,29g DDSA (SERVA) + 28,68g MNA/NMA (SERVA) poured together, and according to GLAUERT: warmed up in an oven to 45 = degrees centigrade, then mixing with a special, selfmade glass stirrer, = after some minutes "red color" will appear, mixing as long as the = yello-brown color appears, then add your accelerator (DMP-30, SERVA) as e.g 1.35% (w/w) or as 1.75% = (w/w), respectively, according to Luft=B4s recommendation, to add 1-2% = of accelerator DMP-30. The color of the resin during and after mixing shall be yellow-brown; = polymerization in our lab is classically: 37, 45, 60-64 degrees C, ~ = 12-24 h (over night) each. If we have to perform "fast and hot" = polymerization, we infiltrate tissue specimens (up to 3 times for at = least 1 h/each) in and embed then specimens into a resin mixture = containing 2% (w/w) DMP-30 (acc. to LUFT). This modified resin recipe essentially is a "recalculated" w/w-mixture = of LUFT=B4s A:B-composition, in a relation of A:B =3D 3:7, which yields = polymerized blocks of a higher hardness, suited for human pathological = specimens (also large specimen cutting areas up to 4 x 5 mm) as we = process them now for about 15 years without problems. 5) To Dr. GARBER: we have met several times (I think) at congresses in = Europe as well as in the USA. Unfortunately I don=B4t know anything = about an "MCEM 98 meeting next week at Portoroz, YU", you mentioned. A = copy "directions for the use of SPI-Pon 812" I greatly should appreciate = either via this e-mail server or via my fax indicated below. Thank you = also for the informations on how to order saving 30% of the costs... Thank you very much for responding and your help,=20 other opinions or informations by any colleague out there on the subject =
(anyone out there, performing the two-step polychromatic staining = procedure of HUMPHREY and PITTMAN 1974: Azure II-Methyleneblue-basic = fuchsin?? on Epon 812 or EPON 812-replacement resins?? handling, = results, satisfaction?) are warmest welcome.
best regards=20 Wolfgang MUSS EM-Lab of Dept. Pathology, LKA Muellner Hauptstrasse 48 A-5020 SALZBURG, AUSTRIA/Europe phone: ++43++662-4482-4720 Ext Fax: ++43++662-4482-882 Ext (c/o:W.MUSS) E-mail: W.Muss-at-lkasbg.gv.at END of message
Although I have commercial interest in this subject, I am mentioning it because the discussion of archiving systems is already out there, with specific systems listed...I mention our system as an alternative to those being discussed.
BUEHLER, LTD. also sells an image capture/archiving system; the M.A.R.S.(TM) System. M.A.R.S. comes complete with a camera and pentium computer. An optional high resolution (three chip) camera is available.
Features include image capture and enhancement; complete image annotation and editing; measurement tools including: Point to point, Parallel beam, Perimeter/ Area of a polygon, Radius, Angle, and Weld bead angle bisection; Hardness test measurement and Vickers/ Knoop calculation; Image comparison (5 images overlap for grain size comparison, etc.; and automated report generation through a WORD(R) interface. WORD templates come standard, but customized report templates are easily created. Web site info is available at http://www.buehlerltd.com
For more info, you can contact your local salesman or call (800)323-9330 or (847)295-6500.
Scott D. Holt BUEHLER, LTD. PO Box 1 41 Waukegan Rd. Lake Bluff, IL 60044 (847)295-6500 http://www.buehlerltd.com
Dear Reader: Is any one know how to fix and process 17 and 18 days mouse embryo in paraffin embedding? I have no problem to cut 16 days mouse embryo paraffin sections. However, not 17 and 18 days, those tissues are too mushy to cut. They looks like unfixed and wax cann't get into tissue. Thanks.
Dorothy Zhang Harvard School of public Health Building 2, CVLAB 677 Huntington Ave, Boston, MA 02115 Phone# 617-432-2970
} } http://www.lumen.luc.edu/lumen/MedEd/Histo/frames/histo_frames.html } http://www.usc.edu/hsc/med-sch/images/images.html } http://www.kumc.edu/instruction/medicine/anatomy/histoweb/ } } Links to these sites (and others) may be found at: } } http://www.pharmacy.arizona.edu/centers/tox_center/swehsc/exp_path/w3-hist } o.htm The last link does not work! Other may try as well out site which has many teaching modules and = link to other pathological sites. http://www1.omi.tulane.edu/classware/pathology/medical_pathology/overvi= ew.html. My own site has lots of inner ear stuff and it is listed as virtual = resource by the Association for Research in Otolaryngology at site = http://www.aro.org/
*Disclaimer: Whatever... is not Tulane =B9s opinion! Cesar D. Fermin, Ph.D. Professor of Pathology & Otolaryngology web: http://www.tmc.tulane.edu/ferminlab, Internet: = fermin-at-tmc.tulane.edu 1430 Tulane Ave/SL79 New Orleans, La 70112-2699 Fax 504 587-7389, Voice 584-2521, Main Office 588-5224
THE NEW ENGLAND SOCIETY FOR MICROSCOPY (NESM) will hold its October Meeti= ng at M/A-COM, Incorporated 1011 Pawtucket Boulevard, Lowell, MA, (508) 442-4400 on Wednesday, Octobe= r =
22, 1997 at 5:00 PM.
PROGRAM
5:00 pm Registration
5:10 pm Tours Note: 40-minute, escorted tours of the M/A-COM labs will=
start from the NESM registration area, leaving every 20 minutes (or as groups of 10 arrive) until around 6 pm. =
=
6:00 pm Buffet Dinner =
7:15 pm "How to Find Out Why" (Case studies illustrating the use of vario= us tools, including microscopy tools, to determine how and why microelectron= ic components fail) presented by Dana Crowe, Manager of Corporate Reliabilit= y Engineering, M/A-COM, Inc. =
=
8:00 pm "Endocytosis in Alveolar Epithelial Type II Cells" (A discussion = of potential pathways by which ultrafine particles cross the lung epithelium=
and enter the pulmonary interstitium) presented by Rebecca Stearns, Dept.=
of Environmental Health, Harvard School of Public Health =
=
NEW MEMBERS WELCOME!
To register, contact L. Kirstein at tel: 508-473-9673 or E-mail:104365.3522-at-compuserve.com. =
Cesar & others, Actually the URL was missing the "l" on .html part and I always have problems when my URLs are sent via email because the URLs that they allow me at the College of Pharmacy are so doggone long that they wrap to the next line [yes I've tried to get them to alias the sites, but it falls on deaf ears (sorry Cesar ;-) ]. Try using http://www.pharmacy.arizona.edu/exp_path.html and "drilling down" to get to the site.
BTW Cesar, the Tulane site you referenced looks like a good one.
Doug Cromey
At 12:03 PM 10/1/97 +0000, you wrote: } } http://www.lumen.luc.edu/lumen/MedEd/Histo/frames/histo_frames.html } } http://www.usc.edu/hsc/med-sch/images/images.html } } http://www.kumc.edu/instruction/medicine/anatomy/histoweb/ } } } } Links to these sites (and others) may be found at: } } } } http://www.pharmacy.arizona.edu/centers/tox_center/swehsc/exp_path/w3-hist } } o.htm } The last link does not work! } Other may try as well out site which has many teaching modules and link to other pathological sites.=20 } http://www1.omi.tulane.edu/classware/pathology/medical_pathology/overview.h tml. } My own site has lots of inner ear stuff and it is listed as virtual resource by the Association for Research in Otolaryngology at site http://www.aro.org/ } } *Disclaimer: Whatever... is not Tulane =B9s opinion! } Cesar D. Fermin, Ph.D. Professor of Pathology & Otolaryngology } web: http://www.tmc.tulane.edu/ferminlab, Internet: fermin-at-tmc.tulane.edu = =20 } 1430 Tulane Ave/SL79 New Orleans, La 70112-2699 } Fax 504 587-7389, Voice 584-2521, Main Office 588-5224 } } ..................................................................... : Douglas W. Cromey, M.S. Dept. of Cell Biology & Anatomy : : Sr. Research Specialist University of Arizona : : (office: AHSC 4212A) P.O. Box 245044 : : (voice: 520-626-2824) Tucson, AZ 85724-5044 USA : : (FAX: 520-626-2097) (email: doug-cromey-at-ns.arizona.edu) : :...................................................................: http://www.pharmacy.arizona.edu/exp_path.html Home of: "Microscopy and Imaging Resources on the WWW"
In light of Alwyn's upcoming departure from the University of Illinois a Web Page has been prepared for Alwyn on which you can record messages and regards for those of you who are unable to participate in person. The address is http://ginny.mrl.uiuc.edu/alwyn/ .
Roxanne Luesse Administrative Secretary Materials Research Laboratory 104 S. Goodwin Urbana, Illinois 61801 Phone: 217-333-1370 Fax: 217-244-2278
I published a paper on a modification of the TOTO technique which works well on plants. The reference is: FOOD STRUCTURE, Vol.12 (1993) pp.475-482. If you wish to talk to me about it, feel free to contact me directly.
Bill
William R. McManus Electron Microscopy Facility Department of Biology Utah State University Logan UT 84322-5305 1-801-797-1920 billEMac-at-cc.usu.edu
Wolfgang Muss wrote: ================================================ Unfortunately I don´t know anything about an "MCEM 98 meeting next week at Portoroz, YU ================================================ Acutually it is Portoroz, Slovenia, the dates are October 5-8, and information about it can be found by looking at the SPI website under "Hot Meetings". There is still time to register, contact person is Dr. Miram Ceh, E:mail {mcem97-at-ijs.si} .
Let me give a sales pitch for this meeting, my only "benefit" will be that maybe you might visit our SPI exhibit stand and say "hello":
1] You will be able to hear an outstanding lecture by "our own" Nestor Zaluzec on the topic of "Analytical Electron Microscopy and Materials Research at ANL". It is also my understanding that there is going to be some kind of round table discussion on the "future" in terms of internet communications between scientists in microscopy.
In fact, the quality of the scientific program is outstanding. Another "Plenary lecture" will be "Pigments of the Gastrointestinal Tract.A Saga of Botched Histochemistry" by Feroze N. Ghadially. There is an impressive number of poster presentations on virtually all areas where one uses EM.
2] You really should check out the MCEM website, this is a great meeting about to happen! And if you have have never been to that part of the world, I am told that there is nothing like visiting the old port city of Portoroz!
3] Portoroz is not more than a day's drive from many of the major capitals of Europe!
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I believe you are refering to the OTO method described in the article "Thiocarbohydrazide-mediated osmium binding: a technique for protecting soft biological specimens in the scanning electron microscope" by Robert O Kelly, etal. in Principles and Techniques of Scanning Electron Microscopy. vol 4. 1975. ed by M.A.Hayat. ISBN 0-442-25686-8. This method enhances osmium binding and therefore reduces charging and heating of difficult to gold -coat structures. I am not sure how well this would work with plant material. Of course you may be refering to another method.
Hank Adams New Mexico State University Las Cruces, NM
My specimen holder in our Polaron E3000 CPD is not permitting the intermediate fluid to drain out the bottom, ie the tiny hole is blocked. I have removed the retaining screw and spring, but the slider rod is stuck. I have tried gently tapping it from bothh ends, but it will not budge.
Before I really try to sledgehammer it, I thought I would ask if it comes out of one end preferentially, ie from the end withh the retaining spring or the opposite end where it fits over the peg in the CPD itself. I'm assuming it is of uniform diameter with no t shoulders, but don't want to risk ruining it totally. I am soaking it in WD 40 overnight to see if that loosens it any.
You might prefer to email me directly. Thanks
Carolyn J. Emerson email: cemerson-at-plato.ucs.mun.ca
Biology Department Memorial University St. John's, NF A1B 3X9 Tel: (709) 737-7515 Fax: (709) 737-3018
We have had a rash of incorrect postings to the listserver, lately. Most fall into the following 3 categories...
1.) Incorrect address for subscribe/unsubscribe..
To subscribe and/or unsubscribe you must post your message to :
Listserver-at-MSA.Microscopy.Com
and you must provide your ORIGINAL subscription address
Please do not send subscribe/unsubscribe messages to the list ( Microscopy-at-MSA.Microscopy.Com) . If you can't remember what to do then use our WWW form at:
Also all subscribe/unsubscribe processing is done only once/day. So there is generally at least a 24 hr lag time before you are removed from the server.
2.) Attempts to Post messages but to the wrong address..
To post a message send it to:
Microscopy-at-MSA.Microscopy.Com
several people have been posting messages to the server Administrative Address (Listserver-at-MSA.Microscopy.Com) which I must then redirect manually. This obviously wastes time and effort!
3.) Advertising.
Please remember our rules about Advertising (i.e. it is not allowed) there have been numerous "violators" lately many of which are walking in the grey area of "just" going over the line and most of those individuals have received an electronic slap on the hand from me. However, there has been one repeat offender who has been removed from the list as they continued to violate our rules after repeated requests to stop. If you can't remember the rules they are, of course, posted on the WWW
Please note that for nearly all of next week I will be "off-line" and the system will be on auto-pilot. I EXPECT that there will be glitches in the subscribe/unsubscribe functions as at least 25% of all of the unsubscribe messages have some type of an error (how many ways do you think people spell unsubscribe?.. all of those must be manually processed !) Unfortunately, my links to the NET will be pretty minimal to non-existant next week. So please be patient, or plan ahead and unsubscribe before Friday Oct 3.
I have a little problem with a selective colour revealing the microstructure of the Ti6Al4V alloy - this is a wax-lost cast. The gas bubbles appear when etching by imersion a specimen in a water solution of the NH4F.HF reagent so a thin film of the reaction products cannot deposite on a ground face of a specimen. Finaly I receive the black-and-white results of etching.Do you know the reason of these bubbles creation or maybe you know some other etchants withoght NH4F.HF reagent to tint a microstructure of Ti-Al-V alloys ?
Best regards for all
Janina Radzikowska e-mail: jradz-at-iod.krakow.pl Foudry Research Institute Krakow, Poland
I have a little problem with a selective colour revealing the microstructure of the Ti6Al4V alloy - this is a wax-lost cast. The gas bubbles appear when etching by imersion a specimen in a water solution of the NH4F.HF reagent so a thin film of the reaction products cannot deposite on a ground face of a specimen. Finaly I receive the black-and-white results of etching.Do you know the reason of these bubbles creation or maybe you know some other etchants withoght NH4F.HF reagent to tint a microstructure of Ti-Al-V alloys ?
Best regards for all
Janina Radzikowska e-mail: jradz-at-iod.krakow.pl Foudry Research Institute Krakow, Poland
Janina.Radzikowska wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello! } } I have a little problem with a selective colour revealing the } microstructure of the Ti6Al4V alloy - this is a wax-lost cast. } The gas bubbles appear when etching by imersion a specimen in a } water solution of the NH4F.HF reagent so a thin film of the reaction } products cannot deposite on a ground face of a specimen. Finaly I receive } the black-and-white results of etching.Do you know the reason of these } bubbles creation or maybe you know some other etchants withoght NH4F.HF } reagent to tint a microstructure of Ti-Al-V alloys ? } } Best regards for all } } Janina Radzikowska e-mail: jradz-at-iod.krakow.pl } Foudry Research Institute } Krakow, Poland
Janina,
Here is some etchants for color etching of titanium and alloys (from http://www.kaker.com/etch/demo/etch.html):
Material: Titanium alloys (Ti) Type: Microetching Method: Physical etching Etchnant: Thermal etching Procedure: The specimen is polished and tinting removed from bakelite holder. Heating is done in a stainless steel or tungsten-filament basket in air. Basket temperature 1200 C (2192 F) approx. Specimen temperature approx. 400-600 C (752-1112 F). Time approx. 15-60 s. Air blowing improves both oxidation rate and time and specimen's temperature control. Remarks: Color etching. Different coloring of the matrix and the various phases is obtained. In titanium-aluminides-based alloys, the TiAl matrix usually appears yellow and brown. The Ti3Al phase usually appears blue or green. Reference: E. Beraha and B. Sphigler, Color Metallography, American Society for Metals (ASM), Metals Park, Ohio 440073, USA, 1977. p. 111.
Material: Titanium alloys (Ti) Type: Microetching Method: Chemical etching Etchant (electrolyte): 3 g ammonium bifluoridie and 4 ml hydrochloric acid (25 %) in 100 ml distilled water. Procedure: Immersion at room temperature for a few sonds. (To obtain good coloring, last polishing stage should be carried out in, or with, saturated solution of oxalic acid.). Remarks: Color etching. Alpha titanium grains are differently colored. sondary alpha and alpha-prime and various intermetallic phases (such as Ti2Cu) remain white (uncolored) or are evident by coloring. Reference: E. Beraha and B. Sphigler, Color Metallography, American Society for Metals (ASM), Metals Park, Ohio 440073, USA, 1977. p. 111.
Material: Titanium alloys (Ti) Type: Microetching Method: Chemical etching Etchant (electrolyte): 5 g ammonium bifluoride in 100 ml distilled water. Procedure: For pure titanium, immersion at room temperature for a few sonds. Longer etching time required for titanium alloys. Remarks: Color etching. Titanium alpha grains and twins are differently colored according to their crystallographic orientation. sondary phases are evident by coloring. Reference: E. Beraha and B. Sphigler, Color Metallography, American Society for Metals (ASM), Metals Park, Ohio 440073, USA, 1977. p. 111.
Material: Titanium alloys (Ti) Type: Microetching Method: Chemical etching Etchant (electrolyte): 2-3 g sodium molybdate, 5 ml hydrochloric acid (35 %) and 1-2 g ammonium bifluoride in 100 ml destilled water. Procedure: Immersion at room temperature until specimen surface occurs. Remarks: Color etching. Etching of as-cast titanium alloys.Titanium alpha matrix is colored blue or green. TiC is colored yellow or dark brown. Reference: E. Beraha and B. Sphigler, Color Metallography, American Society for Metals (ASM), Metals Park, Ohio 440073, USA, 1977. p. 111.
Henrik
-- Henrik Kaker SEM-EDS Laboratory, Metal Ravne d.o.o. Koroska c.14, 2390 Ravne, Slovenia Tel: +386-602-21-131, Fax: +386-602-20-436 SEM-EDS Laboratory Web Site: http://www2.arnes.si/guest/sgszmera1/index.html Microscopy Vendors Database: http://www.kaker.com/mvd/vendors.html Kaker.Com: http://www.kaker.com
} Let me give a sales pitch for this meeting, my only "benefit" will be that } maybe you might visit our SPI exhibit stand and say "hello": } } In fact, the quality of the scientific program is outstanding. Another } "Plenary lecture" will be "Pigments of the Gastrointestinal Tract.A Saga of } Botched Histochemistry" by Feroze N. Ghadially. There is an impressive } number of poster presentations on virtually all areas where one uses EM. } } Chuck }
Also, revised versions of papers presented at MCEM 97 will appear as a special issue of Journal of Computer Assisted Microscopy, early next year.
} } Is any one know how to fix and process 17 and 18 days mouse embryo in paraffin embedding? I have no problem to cut 16 days mouse embryo paraffin sections. However, not 17 and 18 days, those tissues are too mushy to cut. They looks like unfixed and wax cann't get into tissue. Thanks. { {
It could be fixation but it sounds like a combination of both inadequate fixation and processing. Some specifics would help ie: -Type of fixative, length of time in fixative -Are the specimens bisected prior to fixation and processing -What type of tissue processor is being used -What is your processing protocol, reagents, time in reagents, vacuum and temperature applied to which stations in the processor
Feel free to either e-mail me or call.
regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
The magnetic field in our laboratory is too high for the new high resolutio= n=20 SEM we are going to install. I know of one active magnetic field=20 cancellation system (Oxfords). Can anyone tell me if there are other simila= r=20 systems in the market?
Best regards Lars Oestensson Graenges Technology=20
For years we have "airfuged" our fluorescent antibodies at 95 000 rpm before use to eliminate any aggregates which may have formed during storage. Does anyone know if this can (or should) be done with gold conjugated antibodies?
TIA,
Pat Hales McGill University Dept. of Anatomy & Cell Biology hales-at-hippo.medcor.mcgill.ca
This is a multi-part message in MIME format. --------------E32E6228270A2877B26D0CC7 Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit
Carolyn Emerson (by way of Nestor J. Zaluzec) wrote: } } } My specimen holder in our Polaron E3000 CPD is not permitting } the intermediate fluid to drain out the bottom, ie the tiny } hole is blocked. I have removed the retaining screw and spring, } but the slider rod is stuck. I have tried gently tapping it } from bothh ends, but it will not budge. } } Before I really try to sledgehammer it, I thought I would ask if } it comes out of one end preferentially, ie from the end withh } the retaining spring or the opposite end where it fits over the peg } in the CPD itself. I'm assuming it is of uniform diameter with } no t shoulders, but don't want to risk ruining it totally. I am } soaking it in WD 40 overnight to see if that loosens it any. } Carolyn, DO NOT TRY TO REMOVE IT FROM THE END WITHOUT THE SCREW. The rod has shoulders on it. Remove it from the screw end. I have the Jumbo model and the holders should be similar. The brass rod is most likely corroded. Good luck Gregory Rudomen S.U.N.Y. Stony Brook University Microscopy imaging center --------------E32E6228270A2877B26D0CC7 Content-Type: text/x-vcard; charset=us-ascii; name="vcard.vcf" Content-Transfer-Encoding: 7bit Content-Description: Card for Rudomen, Greg Content-Disposition: attachment; filename="vcard.vcf"
Here are some replies taken from the "Tips & Tricks" archive I maintain. If you would like to check out the whole thread, got to the web address at the end of this message and follow the "Tips & Tricks" link. From there go to "SEM" and look fo the link dealing with fields. Good luck.
The guy I had contact with for the field compensation system at
Linear Reasearch Associates (it's a small company) is Curt Dunnam
(crd4-at-cornell.edu). He's the chief engineer, and will likley be happy to
talk.
Ben Simkin (simkin-at-egr.msu.edu)
simkin-at-egr.msu.edu
I've had similar problem with our microscope, and if it's truly fields,
I'd recommend just living with not using the outlet. AC fields can be activly
supressed if they are from an external source (we have purchased a system
from Linear Research Associates; they run ads in Microscopy Today (I don't
have the address with me right now)), but this sounds much more like a ground
loop, and the solutions to that (so I've been told) include either sinking
your own dedicated common ground (anywhere from easy to nearly impossible), or
disconnecting the ground connection between your asscessories and your SEM,
and running a "floating ground". I've discussed this as one solution with
our SEM service technician, and he says it sometimes works (assuming the
noise it adds to your instrument signal from the differance in "ground"
potentials is acceptably low).
Ben Simkin (simkin-at-egr.msu.edu)
Dept. Mat. Sci. and Mech.
Michigan State University
Incidentally, there was a series of articles in Microscopy Today (Don Grimes,
Ed., MicroToday-at-aol.com) recently on magnetic fields, their causes and
cures. It was written by Curt Dunnam of Linear Research Associates,
Trumansburg, NY 14886 (Fax: 770-368-8256). LRA apparently specializes in
diagnosing and dealing with magnetic fields in EM labs. You might get useful
help from them. I know there are other companies that do the same, but I
don't happen to have names and addresses readily available.
Wil Bigelow
Wil_Bigelow-at-mse.engin.umich.edu
Hello all,
thanks to all of you who have answered my questions. Here is the relevant
information:
1.1. Two addresse are quoted, maybe the first one is more recent:
THE DINDIMA GROUP P/L
10 Argent Place
RINGWOOG, Victoria, 3134
Australia
Telephone Number +61 3 9873 4455
AND/OR
Post Office Box 106
VERMONT,
VIC 3133
AUSTRALIA
PHONE;
+613-9873-4455
FAX:
+613-9873-4749
1.2. Email is: 100241.3642-at-compuserve.com.au
1.3. If you are in USA, better deal with Chuck (cgarber-at-2spi.com), he has
already done for you the custom process and distributes Arlunya products.
1.4. If you are in Pakistan or nearby country then it might be more useful
2. "Lupe" (actually binocular, sorry for the mistake) I have got several
interesting answers and will answer them privately.
At 02:38 PM 10/2/97 +0100, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 352-392-1295 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
There is company based in USA that can provide an active field cancellation system, that may suit your needs.
address:
Integrated Dynamics Engineering Inc. 150-P New Boston Street Woburn MA 01801 USA
tel: (617) 938-5120 fax: (617) 938-5122
Disclaimer: I have no affiliation with this company, just a satisfied customer.
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
******************************************************** On Thu, 2 Oct 1997 lars.oestensson-at-techno.graenges.se wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } The magnetic field in our laboratory is too high for the new high resolution } SEM we are going to install. I know of one active magnetic field } cancellation system (Oxfords). Can anyone tell me if there are other similar } systems in the market? } } Best regards } Lars Oestensson } Graenges Technology }
We are a lab that does muscle and nerve biopsies, both histology and TEM. We would like to know if there are any atlas-type reference books available of TEM for muscle and nerve (possibly both normal and disease-state). We are interested in using it for teaching purposes (both lab personnel and MD residents).
Thanks in advance, Susan Danielson Medical College of WI Muscle/Nerve lab Milwaukee 414-259-3836
At 09:34 AM 10/2/97 -0700, Pat Hales wrote: } For years we have "airfuged" our fluorescent antibodies at 95 000 rpm before } use to eliminate any aggregates which may have formed during storage. Does } anyone know if this can (or should) be done with gold conjugated antibodies?
Unfortunately, there is no simple answer to this question. Our BioSite gold conjugates are prepared in such a way as to almost totally eliminate clusters (95% of the particles are guaranteed singlets), and centrifuging will actually create, rather than reduce, clumping. However, other gold colloids, produced by other manufacturers, are said to improve with centrifuging. I suggest that you ask the supplier of your gold conjugates for their recommendation.
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
The following is being forwarded to the list as a courtesy. Do Not use E-mail reply function. Thank you. -Karen
Please respond by mail to: Dr. Douglas C. Youvan, CSO KAIROS Scientific Inc. 3350 Scott Blvd., Bldg 62 Santa Clara, CA 95054 USA
dyouvan-at-kairos-scientific.com wrote: } [Below is an]... announcement regarding staff positions that are currently open at KAIROS. The corresponding ad will appear in the October 10, 1997 issue of Science magazine. } } For the software enginnering position, we are especially interested in identifying graduate students or postdocs who are just finishing their studies and who have experience in C++ programming. } } Thanks, } } Doug } } ***************************************************************** ************************** } BASIC + APPLIED R&D POSITIONS } } KAIROS is integrating optical design and software engineering with molecular genetics to develop novel instrumentation, reagents, and methodologies in the fields of biotechnology, microscopy, medicine, and materials science. } } Successful candidates should relate their training and work experience to the content of our website: } } www.kairos-scientific.com } } KAIROS has three immediate openings for scientists and engineers trained in one or more of the following fields: } } Software Development (C++/MFC) } Optical Spectroscopy and Microscopy } Protein Engineering and Cell Biology } } Curriculum Vitate and names of references should be mailed to: } } Dr. Douglas C. Youvan, CSO } KAIROS Scientific Inc. } 3350 Scott Blvd., Bldg 62 } Santa Clara, CA 95054 USA } } Please do NOT respond via e-mail. } ***************************************************************** ************************** } -- } Douglas C. Youvan, Ph.D. } Chief Scientific Officer } KAIROS Scientific Inc. } Bldg. 62, 3350 Scott Blvd. } Santa Clara, CA 95054 } } T: 408-567-0400 x11 } F: 408-567-0440 } W: http://www.kairos-scientific.com } } Editor, Biotechnology et alia http://www.et-al.com
You are dealing with a whole set of interrelated, complicated problems. Briefly- Avoid useing the old Epon from Shell. Noone has it anymore, and if they do, they will not use it because of its unreliablilty between batches. (It was known to contain up to 13% of chlorine left over from the manufacturing process at one time). Noone will be able to duplicate your results, if you use the old Shell Epon in your publication. Use the replacements which are known to be the purified chemical equivalents and are not mixtures of Araldite, Epon, various dilutents (as some of them tend to be). Use LX-112, or Eponate 812.
The interaction between methylene blues and azures, etc. and epon sections are far more complicated than suspected. The coloration depends on osmium penetration (to some extent), on the percentage of unpolymerized monomers left in your tissue, etc. If you would send me your address via e-mail, I will send you a copy of my standard stain (methylene blue, azure, basic fuchsin) method which was a handout at an MSA meeting some years ago. Also you must pay close attention to your stains - are you buying the correct CI number? Have you changed suppliers?
I am short on time right now, but if you contact me with specific questions I would be happy to tell you what I know.
We are trying to do TEM (thin sections) on large liposomes containing cholesterol and triolein (triglyceride) droplets, fixing in solution at room temp then using freeze-substitution & Lowicryl HM23 to minimize extraction of lipids. Ruthenium tetroxide seemed to give better preservation of membrane structure than osmium did, but I don't see the triolein droplets any more. With osmium the triolein droplets are a nice uniform grey. With ruthenium there are dark structures that may be triolein droplets, but they are not homogeneous grey, rather they look like myelin figures, i.e. look like tightly wrapped layers of onion skin (phospholipid membrane?), dark with many concentric thin white lines (railroad tracks?). Particularly odd is that the thin white lines in these dark structures always look sharp as if the membranes were cut perpendicularly, but there is just no way we could always be cutting them perpendicularly.
Does anyone have experience with ruthenium and triglyceride? Is this what you would expect, or does triglyceride stained with ruthenium normally look similar to TG stained with osmium? (We are not experienced at processing liposomes, and these in particular are very sensitive to the process used for EM, in case you have any suggestions.)
I have some color photographic prints of blue fluorescence and yellow fluorescence. Does anyone know how well B&W photos of these prints will turn out? Are there any special filters/tricks that I could try?
TIA
Bob
Dr. Robert R. Wise Department of Biology University of Wisconsin-Oshkosh Oshkosh, WI 54901
(920) 424-3404 tel (920) 424-1101 fax wise-at-uwosh.edu
You can get the same effect as that provided by the large, bulky, and expensive revolving darkroom doors rather more simply by using two doors, separated by a small (3 ft) entry way chamber. Instead of solid doors, however, use heavy black curtains. For each of the doorways use two curtain panels that are fastened at the top and along opposite sides of the door frame, but which overlap by about 18 inches down the middle of the doorway. With this arrangement, you can walk through the curtain panels of the first doorway into the entry chamber. The curtain panels of the first doorway will close behind you, and then you simply walk through the panels on the sedcond doorway into or out of the darkroom.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
From my experience with moving color photos over to B&W, the best way is to bring them into something like Adobe Photoshop and manipulate the grey scale. Blue and Yellow should respond well to this. Photoshop has a control called "Levels" which gives a histogram of grey levels then lets you set where you want the black, white and the midpoint to be within that histogram. I don't know what your pictures look like but if you had a print of a blue image on a black background, for instance, you could even adjust this so that the blue shows up white against the black, both being "pure" black and white if you desire (e.g. 0 and 256). Sometimes this is better than simply adjusting the "brightness" and "contrast" controls. There is also a similar adjustment called "curves" but to be honest, I never seem to get what I want out of that control (most likely due to ignorance on my part).
I hope this helps. Feel free to contact me off line if you need additional info.
Cheers,
John V. Pacific Northwest National Laboratory Richland, WA USA js_vetrano-at-pnl.gov (509)372-0724
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have some color photographic prints of blue fluorescence and yellow fluorescence. Does anyone know how well B&W photos of these prints will turn out? Are there any special filters/tricks that I could try?
TIA
Bob
Dr. Robert R. Wise Department of Biology University of Wisconsin-Oshkosh Oshkosh, WI 54901
(920) 424-3404 tel (920) 424-1101 fax wise-at-uwosh.edu
Dear Bob: The biggest problem is to have the greys represent, light areas as fluorescing. I expect that the yellow will be no problem but the blue will probably be too dark. I would rephotograph the prints in B&W film using colour filters. If you remember the Oswald colour circle which has complimentary - opposite colours one can easily determine which colours will result in lighter or darker greys. So blue, photographed using a blue filter will result in a lighter grey. Orange in a darker. You can achieve the same with digital imaging and colour replacements. Regards Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au } } I have some color photographic prints of blue fluorescence and yellow } fluorescence. Does anyone know how well B&W photos of these prints will } turn out? Are there any special filters/tricks that I could try? } } TIA } } Bob } } Dr. Robert R. Wise } Department of Biology } University of Wisconsin-Oshkosh } Oshkosh, WI 54901 } } (920) 424-3404 tel } (920) 424-1101 fax } wise-at-uwosh.edu } }
} The magnetic field in our laboratory is too high for the new high resolution } SEM we are going to install. I know of one active magnetic field } cancellation system (Oxfords). Can anyone tell me if there are other similar } systems in the market? } } Best regards } Lars Oestensson } Graenges Technology
Hi Lars,
Besides the systems already mentioned by Scott and Fred, there is a system named EMF-1 produced by Advanced Research Systems in Illinois. You can find information about the system on the web-address:
http://www.mcs.net/~ars/ars/emf-1.htm
I have no experience with the system - I just came across this webpage some time ago.
Best regards, Joergen.
J. B. Bilde-Soerensen Senior Research Scientist, Ph. D. Materials Research Department Risoe National Laboratory DK-4000 Roskilde Denmark
Dear Susan, one reference for that: W.J.Kenneth CUMMING, FULTHORPE J., HUDGSON P., MAHON M (Eds): Color Atlas of MUSCLE PATHOLOGY MOSBY-WOLFE (London) 1994, ISBN 0 7234 2016 5, iii-vi, 1- 202 (incl. = subj. index), ~ 140.- US-Dollars is, as to my knowledge, one of the most recently published color atlas, = dealing with Histology, Histochemistry, EM and a bit of Molecular = Biology in Muscle (+/- nerve/nerves not representatively included). For Nerve interpretation/concerning musculature you could look for:=20 a) RICHARDSON E. P. Jr., DeGIROLAMI U. (Eds) Pathology of the Peripheral = Nerve, (+/- TEM, B/W, also histology); Vol. 32 in the Series: Major Problems in Pathology (LiVOLSI Virginia, Consulting Editor). W.B. SAUNDERS (Philadelphia, London...) 1995, 1-164, incl. subj.index; = ISBN0-7216-3298-X (ordered from HARCOURT-BRACE-IOVANOVICH, London, = GB-Pounds ~46.- =3D ~58-60.- US-Dollars) b)VITAL C., VALLAT J.-M.(Eds) Ultrastructural Study of the Human = Diseased Peripheral Nerve, 2nd Ed., ELSEVIER, N.Y.,1987, 1- 286, incl. = subj. index; ISBN 0-444-01136-6, ~165.- US-Dollars
All three books are (in my opinion) worth their price; they are used = also for interpreting muscle cases. Hope this helps, best wishes and have a good day Wolfgang MUSS, A-5020 SALZBURG/Austria e-mail: W.Muss-at-lkasbg.gv.at.
I would be interested to hear from other SEM users with the same (JOEL JSM 5400) or a similar SEM what sort of filament life you are achieving ?
I fear that we may have a vacuum leak since the filament life is characteristically low and tarnishing is usually visible on the filament holder. The latter I am told may be an indication of a vacuum leak in the system.
3rd Oct.97 Dear all, today I got the data sheets (in English, as well as contact adress of = the selling company in USA) on the silicone rubber product I use for = long time. So I am able to send within the next 3-4 days my infos on how to produce = home-made silicone rubber embedding molds to all colleagues out there = who wanted to get that informations.=20 For those who wish/wished only e-mail info on the technique: I shall = contact you directly by e-mail within the next days.=20 A short version of how to plan/to do it, what type of silicone rubber I = use, technical hints and considerations, especially for posting in the = archives (to Scott WHITTAKER) is considered to be prepared next week.
Wish good luck to all=20 (hopefully my technique helps to solve problems. I should be glad = receiving a feedback, if anyone will try the whole.
Note added: I am/was producing and selling (at prime cost + 10% + postage) such = molds for some european as well US- colleagues who don=B4t/didn=B4t want = to fabricate their own negative-forms (which is the most expensive and = time-consuming part remembering that you should be able to produce not = only one or two, but many molds from them, but, a very nice part for = "left-brains") for conventional TEM-embeddings of = epoxide-resins/polyester-resins in following types: - cubic blocks, numbered 1-48 - cubic blocks, numbered 1-12, dito, numbered 1-10 - pyramidal, size + 0 - mold for specimen infiltration consisting of 7 by 4 rounded cavities, = ~ 1.5 ml resin / each cavity. If you want more information on that, feel free to mail to me your = questions or queries. =20 Disclaimer:=20 I have no financial interest in nor am affiliated to the company selling = the silicone rubber product and only speak as a satisfied customer.
Wolfgang MUSS, PhD. Dept. Pathol., LKA, EM-Lab Muellner Hauptstrasse 48 A-5020 SALZBURG, AUSTRIA phone: ++43++662+4482-4720 Ext. fax: ++43++662+4482-882 Ext (c/o W.Muss) e-mail: W.Muss-at-lkasbg.gv.at (character next right to -at- is a "small" L)
Jurgen Paetz wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Dear fellow SEM users } } I would be interested to hear from other SEM users with the same (JOEL } JSM 5400) or a similar SEM what sort of filament life you are achieving } ? } } I fear that we may have a vacuum leak since the filament life is } characteristically low and tarnishing is usually visible on the filament } holder. The latter I am told may be an indication of a vacuum leak in } the system. } } Regards } } J. Paetz (Senior mineralogist) } } Amplats Research Center } Republic of South Africa
Dear Jurgen Paetz,
You are correct that a discolored base is usually a sign of a poor vacuum. A normal burnout of a filament should have a bubble on top of the broken wire. If your filament burns out this way and the base is discolored it is probably a bad vacuum. If the filament is craked , no bubble, then their could be a flaw in the wire or a slight crack was made by human handling. We do not use a JEOL ourselves, but we do sell filaments for all the different scopes and our customers seem to feel you should get 50 - 200 hours out of a filament.
I have the same machine as yours with selected operation for low vacuum chamber (JSM-5400LV). For high vacuum operation, more than 100 hours, sometime 130 hours of filament life can be achieved.
****************************************** Zhiyu Wang Electron Microscope Lab and Imaging Center Western Kentucky University(WKU) Bowling Green KY 42101
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear fellow SEM users } } I would be interested to hear from other SEM users with the same (JOEL } JSM 5400) or a similar SEM what sort of filament life you are achieving } ? } } I fear that we may have a vacuum leak since the filament life is } characteristically low and tarnishing is usually visible on the filament } holder. The latter I am told may be an indication of a vacuum leak in } the system. } } Regards } } J. Paetz (Senior mineralogist) } } Amplats Research Center } Republic of South Africa } } }
We have a position open for an EM technician.(see below) Informal enquiries can be made to me cgilpin-at-man.ac.uk
UNIVERSITY OF MANCHESTER SCHOOL OF BIOLOGICAL SCIENCES
RESEARCH TECHNICIAN, GRADE C (ELECTRON MICROSCOPY)
Applications are invited for the position of electron microscope technician within the School of Biological Sciences' Electron Microscope, Graphics and Photography Unit. The post will involve working closely within a team of technicians within the Unit in providing assistance with electron microscope operation, sample preparation, image processing, image archiving, data interpretation, networking and computer software management and maintenance. Applicants should have working experience in electron microscopy and preferably a working knowledge of computers. The salary for this post is stlg11,365 p.a.
Application forms are available from Mr. A. Nicholas, School of Biological Sciences, University of Manchester, 2.205 Stopford Building, Oxford Road, Manchester M13 9PT UK. arthur.nicholas-at-man.ac.uk The deadline for applications is October 31, 1997.
The University of Manchester is an equal opportunities employer. Chris Gilpin Biological Sciences Electron Microscope Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 161 275 5170 fax +44 161 275 5171 http://www.biomed.man.ac.uk/biology/emunit/emhome.html
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have had all of my nerve and muscle questions answered in; Myology by Andrew G. Engel and Betty Banker, Vol I and II, McGraw-Hill. I have no idea of the year of the latest edition or the price. My edition (1986) has 2106 pages. Kate Connolly
We are looking for a Reichert-Jung FC4E cryochamber and its control unit for our Reichert-Jung Ultracut E ultramicrotome. If you have an FC4E that you'd like to sell, please contact me.
Russell E. Cook Scientific Associate Electron Microscopy Center for Materials Research Argonne National Laboratory Building 212 9700 South Cass Avenue Argonne, IL 60439 (630)252-7194 FAX: (630)252-4798
Bob: The trick is to use Kodak Panalure II Repro RC paper which is designed for making B&W from color negatives. Tom
} I have some color photographic prints of blue fluorescence and yellow } fluorescence. Does anyone know how well B&W photos of these prints will } turn out? Are there any special filters/tricks that I could try?
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
} I have some color photographic prints of blue fluorescence and yellow } fluorescence. Does anyone know how well B&W photos of these prints will } turn out? Are there any special filters/tricks that I could try? } } TIA } } Bob } Bob,
They should turn out fine. I take it you're meaning to take copy-stand photos of the prints? Just use the filters as if you're taking B&W negatives of the original preps. Yellow 12 filter to lighten the yellow and darken the blue, a blue filter for the reverse, a red to really darken the blue.
Or no filter. If the blue areas are nicely a blue (not light blue) they should naturally have a darker grey value. Try shooting a roll of a few prints, doing each one no filter, Yellow 12, blue (I forget the number). Maybe even a red.
I assume you're shooting with T Max? Another choice would be to use mumbletymumble, some orthochromatic B&W. This will have a reduced sensitivity to blue, and will shoot as if you were using a yellow filter (more-or-less).
The only other trick is the usual: watch for reflections off of the prints, but if you're using a copystand, this should be moot.
} I would be interested to hear from other SEM users with the same (JOEL } JSM 5400) or a similar SEM what sort of filament life you are achieving } I fear that we may have a vacuum leak since the filament life is } characteristically low and tarnishing is usually visible on the filament } holder. The latter I am told may be an indication of a vacuum leak in } the system.
How long is your filament lifetime?
I work on a CamScan SEM/EDX and have lifetimes from 90-240 hours, depending on how much EDX (20kV, SEM: 15kV) I made. The lifetime of the filament does not seem to depend on the sort of the filamnet, cheap ones have similar lifetimes to more expensive ones.
Our SEM has no lock, so we have to ventilate (with N2) the whole column, including the filament.
Greeings, O.Rother -- Oliver Rother Institute for Geology and Paleontology Scanning Electron Microscope (SEM) University of Kiel, Germany Tel. +49 431 35021 Fax: +49 431 35262
Reference to the problem with stray magnetic field. We have installed a Field Cancellation System by Spicer Consulting which we purchased from Agar Scientific,Stanstead, Essex, UK in all the labs in which we have high resolution instruents. They cost (3years ago) about 8000 Uk pounds each. They work VERY well.
Parick Echlin Multi-Imaging Centre University of Cambridge
On Thu, 2 Oct 1997 lars.oestensson-at-techno.graenges.se wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } The magnetic field in our laboratory is too high for the new high resolution } SEM we are going to install. I know of one active magnetic field } cancellation system (Oxfords). Can anyone tell me if there are other similar } systems in the market? } } Best regards } Lars Oestensson } Graenges Technology }
Rinse grids easily this way: Find a source of freshly, freshly, (not a typo) distilled water. (The collecting jars for the water must be maintained cleanly, as well as the tubing, and the water must not "sit" for days in the jars). If you must use deionized water be aware that it may not be as clean. That is, deionized water may be filtered through a 3 micrometer filter and then through a 1 micrometer filter. If you then apply a 0.22micrometer filter before use, the water may still not be really clean enough for TEM. Please be aware of all these possibilities if you see miscellaneous dirt on your sections. What most of us do not realize graphically is that a particle which passes through a 0o.22 micrometer filter may be really huge at a 15x mag. At any rate, fill a clean syringe with good quality water. Attach a 0.22 micrometer filter which you have cleaned by running through it (at a previous time) about 15cc of boiling water. (some filters contain dust aquired during the manufacturing process). When rinsing grids allow genrous quantities of water to run down the forceps and over the grids. Blot with dustless filter paper. If we have a lot of grids to do, we use the Hiroka Staining Kit. This works really well and is easy to use, especially if one only loads the center three or four rows of the pad. Most important - always pay attention to your water! It has to be as clean as you can get it. Bye, Hildy
How are the lines applied to TEM fluorescent screens? I have new screen that is blank and would like to apply at least a recognizable spot at the center, maybe a frame outlining the photo area. I'd think that I could use a drafting pen. True?
TIA, Owen
Owen P. Mills Michigan Technological University Metallurgical & Materials Engineering Rm 512 MME Building Houghton, MI 49931 906-487-2002 906-487-2934 FAX opmills-at-mtu.edu
Since I have archived a discussion on how to make your own TEM screens, I would appreciate if you would be so kind as to send me the replies to this thread. It would make a nice addition. Thanks
At 11:41 AM 10/3/97 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 352-392-1295 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
Chris, Terribly sorry chap, but I'm rather confused about the salary you offered in your posting for the technician position. Would you mind translating {stlg11,365 p.a.} into English, American English that is! ;-);-);-) ------------------------------------- Name: Winston W Wiggins E-mail: wwiggins-at-carolinas.org
Fellow Histoneters that responded to running controls on immunofluorescent (IF) studies. The response was not great. Maybe 6. The responses ranged from not running controls at all; to using tonsils, which seemed to appeas the inspectors; to being unrealistic "request that your pathologist or urolgist give you removed kidneys with tumors that can be negative for everything except IgG." As of 9/29/97 our lab had a positive (renal needle bx) lupus, in which I sectioned several blank slides, cold acetone fixed, and am storing in a -30oC. Upon receiving another IF case and tagging as per usual with IgG, IgM, IgA, C3, C1q, K & L, one of the known lupus slide will be tagged with Polyvalent (IgA, IgM, IgG, Kappa, Lambda). Thank you everyone for allyour imput and valuable help. If anyone else has a realistic suggestion, please e-mail me. Teresa
Richard Thrift asked about preservation of liposomes in embedded tissue. For traditional ethanol-dehydrated, epon-embedded preparations for TEM, check out reference by Angermuller and Fahimi from 1982 in Histochemical Journal 14:823-835 on imidazole-buffered osmium tetroxide. They obtained excellent preservation and staining of lipid droplets and lipoproteins with this technique applied to rat liver. Was particularly effective in preserving lipids with unsaturated fatty acids such has the oleic acid in triolein. We had good results in preserving emulsions composed of lecithin, cholesterol, and triolein. Don Gantz Boston Univ. School of Medicine gantz-at-med-biophd.bu.edu
We are in need of some extremely low power objectives (1X, 2.5X and 5X) lens for a Leica light microscope (all brands will be considered). The lenses should have flat field and extremely good resolution cababilities.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing 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 National Institute of Standards & Technology has many Post Doctoral Positions open. These are offered competitively through the National Research Council. Within microscopy/microanalysis research areas we have several possible openings described at the following sites:
http://rap.nas.edu/lab/NIST/50837106.html This opportunity highlights our analytical research using Analytical Electron Microscopy/Compositional Mapping
http://rap.nas.edu/lab/NIST/50837109.html This opportunity highlights our Submicroscopic Chemical and Physical Characterization of Materials and Particles
http://rap.nas.edu/lab/NIST/50837110.html This opportunity highlights our Electron-Probe Microanalysis/Scanning Electron Microscopy research.
These are very general descriptions of broad areas of research. If you have research ideas that are related to these analytical approaches and are looking for a Post Doc opportunity, please contact me soon.
The NIST/NRC program offers a two year post doc at an annual salary of $45,500. The applications are due to the NRC in January 1998. This includes a technical proposal and several recommendations.
A candidate must be a U.S. citizen that receives their PhD and starts work at NIST by Jan 15, 1999 (You can start as early as July 1, 1998). So, this is the perfect opportunity for those of you that are graduating this spring through next fall. PLEASE NOTE that NIST/NRC only accepts applications ONCE a year, unlike some other institutions.
Eric B. Steel, Leader e-mail: eric.steel-at-nist.gov Microanalysis Research Group Office: 301-975-3902 N.I.S.T. FAX: 301-417-1321 Bldg. 222/Rm A113 Gaithersburg MD 20899 http://www-sims.nist.gov/Division/MicroGroup.html
I have used both techniques with varying results. CPD often caused artifacts, but also produced more attractive images than freeze drying. Many cells collaps with FD. My best results were obtained in cryo or with fresh hydrated samples (you get about 30 min. observation time before the cells collaps; depending on what your specimens are, of course). If you have access to an ESEM this may be the best bet.
best wishes
Sincerely +----------------------------------------------------------------- |Dr Stephan Helfer, SSO |Senior Mycologist - MSc Course Director | |Royal Botanic Garden Edinburgh, Inverleith Row, EDINBURGH EH3 5LR, |Scotland UK | |http://www.rbge.org.uk | |phone: +44 (0)131 552 7171 ext 280 | or +44 (0)131 459 0446-280 (direct digital VoiceMail line) |fax: +44 (0)131 552 0382 +------------------------------------------------------------------
"What is the best commercial source of consistently high-quality holey grids?"
This is a seductive question for a vendor to answer?
I am associated with Ted Pella Inc. in Redding, Northern California and would venture to say that our holey films are of consistently high quality.
I am not clear from your question whether you are looking for holey films for astigmatism correction or holey films for specimen application. If the latter - we supply a product we call NetMesh Grids, Lacey films. These contain many holes of different sizes in a netlike pattern and are very strong.
Please contact us at 1(800)237-3526 or 1(808)573-8945.
To everyone in my e-mail address file (sorry if you have this information already): please note that I am taking a new job and moving in about ten days.
ALWYN EADES
(full name: John Alwyn Eades)
IS MOVING TO LEHIGH UNIVERSITY IN OCTOBER 1997
Present address
Materials Research Laboratory 104 S Goodwin Urbana Illinois 61801-2985
is moving to
Department of Materials Science and Engineering Lehigh University Whitaker Laboratory 5 East Packer Avenue Bethlehem PA 18015-3195
610 758 4231 610 758 4244 FAX jae5-at-lehigh.edu not yet activated ** Alwyn Eades Center for Microanalysis of Materials University of Illinois at Urbana-Champaign Phone 217 333 8396 Fax 217 244 2278 eades-at-uimrl7.mrl.uiuc.edu (NB those are letter l not ones)
In mid-October this year (1997), I will be moving to Lehigh. The new address will be:
Department of Materials Science and Engineering Lehigh University Whitaker Laboratory 5 East Packer Avenue Bethlehem, PA 18015-3195
Filament life is a function of many factors. The presence of tarnishing usually signifies some sort of oxidation, implying a vacuum leak. I must say, if your SEM is a new one, that filament life usually improves over the first year of life. I think this is a result of outgassing the whole system. BTW, if you have a "bubble" at the end of your burnt-out filament, this is a result of over-saturating the filament. Remember to check the satuation level every hour for the first six hours of a new filament's life, as the saturation level drops fairly quickly, then levels off. After the second year, a filament lasts me a month.
I do not have a JEOL 5400, these are just general W-filament comments.
You wrote: } Dear fellow SEM users } } I would be interested to hear from other SEM users with the same (JOEL } JSM 5400) or a similar SEM what sort of filament life you are achieving } ? } } I fear that we may have a vacuum leak since the filament life is } characteristically low and tarnishing is usually visible on the filament } holder. The latter I am told may be an indication of a vacuum leak in } the system. } } Regards } } J. Paetz (Senior mineralogist) } } Amplats Research Center } Republic of South Africa } Regards, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
I am impressed with, and use, holey carbon films prepared by Structure Probe. Dozens of grids with these films have been uniformly excellent. Recently I had a chance to use a couple of holey grids made by Pella, and they were excellent also. I don't recall the pricing comparison, but both are cheaper than I can make them myself (and probably better :-) ).
Larry
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
Vachik Hacopian wrote: =================================================== What is the best commercial source of consistently high-quality holey grids? =================================================== As a long time manufacturer of filmed grids, including "holey" and "lacey" types, my answer would not be exactly that of an independent third party.
However, just remember one thing: The "making" of filmed grids is easy to describe, however the "art" of making a superb grid is not. But at the end of the day, no one knows for sure what they have made unless they have their own in-house TEM facilities to inspect the quality of their products. Be certain your intended vendor has their own facilities to check themselves what they are getting ready to send out their door.
Otherwise you, the customer, will be the QC department for that vendor yourself! Anyone who has been unhappy with purchased filmed grids in the past will know exactly what I am talking about. Customers are always welcome to visit our production facility and meet our staff members responsible for the in-house production and TEM inspection of our coated grids.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I have just come back from working in the USA and on reading my e-mail I saw your request and some of the answers provided.
Perhaps I have been in this game too long! I just had to sit down and tr= y to help you out as I feel the answers that I saw were not exactly correct= ? =
We are talking about Tungsten, the life of LaB6 is another question!
1. I can not speak directly for the instruments that you mention but =
ALL SEM are the same when it comes to filament life 1.1 Filament life is inversely proportional to the current you wish to=
draw from the gun, emission current measured in micro amps. 1.2 Typically Japanese instruments need ~100uA for good quality high resolution images, Philips use a lower current than other instruments usually between 30 and 50 uA at saturation 1.3 The position of the filament in the cathode and the bias setting decide the total current that you may pull from the gun. The nearer the filament to the cap the higher the current that you may attain. 1.4 To check to see if you have the "optimum - high performance" filame= nt position run up to 40,000X (working distance 10 to 15mm) with a nice specimen which gives lots of signal and see if you obtain an image (even = if its noisy) through the whole of the spot size (often called probe current= ) range. If you do the filament is fine if you do not the filament is in a= n "economy position" which will reduce current but increase filament life. =
If you need to work at high resolution you will need the filament in the optimum position, for less than 20,000X and for EDX this is not required.=
2. Filament life is also very dependant upon gun vacuum, it is possible = to have a poor gun vacuum even if the vacuum gauge says the vacuum is good. =
The gun will be very smelly in a poor vacuum environment, the chamber tending to be orange in colour and the filament base will also tend towar= ds an orange colour. Filament bases will ALWAYS show a colour, pale blue =3D=
low heating level low current use, dark blue =3D higher heating and more=
normal if the instrument is being used for highish performance, orange-brown =3D contamination through poor vacuum 3. Filament life also depends on how carefully the filament is saturated=
i.e. use a wave form and make sure you fully saturate but do not overheat=
the filament. 4. You should check saturation and alignment every time you switch the k= V on and every time you change the kV. As the filament ages and thins it will require a different current to attain saturation. When you change t= he kV the gun becomes more (up) or less (down) efficient and the saturation will change, on many instruments so will the emission current. 5. Due to the higher currents used in the SEM 90% of filaments failing under normal use will have small "melt" blobs at the failure point, this = is normal. A large melt blob usually means oversaturation. In the TEM the currents drawn are far lower and the usual break is between two tapered ends, blobs are more rare unless oversaturated.
So after all that how long should they last? Well 30 to 50 hours is OK i= f you are running in a normal lab environment. If you are pushing the resolution expect 15 to 30 hours. Aiming at better resolution than the manufacturer claims, then you must expect to have {10 hours filament life=
and you will end up with a very dirty gun and first condenser system; but=
you are getting more than you paid for! If you get well in excess of 50 hours my personal belief is that the instrument is not being used to its best effect, there is far more in a SEM than using it as a super light microscope! Of course it is horses for courses, unfortunately too few people realize just how much performance they could really pull from thei= r SEM if they only asked the instrument in the correct way for more performance. =
Electron microscope operators must realize that the filament is a consumable item! As I travel the world I really believe that the life of=
the filament in most laboratories takes priority over the quality of the image. Every laboratory I visit has the filament in the long life econom= y position and they almost all complain that the instrument will not do wha= t they want. You cannot get good high resolution or low kV results with ou= t plenty of beam current and that starts by setting the gun up correctly an= d in that case you must sacrifice filament life. Claims of 100 hours plus = in a SEM must mean the gun is under run, probably the operator is using the first peak - too big for imaging at any level of performance. Nothing wrong with this but any complaints of poor performance need to be rectifi= ed by first looking at filament position and saturation.
In my books "Working With a SEM" and "Maintaining and Monitoring the Electron Microscope" I discuss filament breaks and the colour of the base=
under different conditions.
Sorry to rabbit on but the problem of being in this game for 33 years is that you have seen it all before! It is so frustrating that SEM are considered by many people to be instruments to look at flies eyes or bee'= s knees, when they are really a scientific instrument with a very complex imaging system and tremendous potential when used correctly. You should = be able to run even a 10 year old instrument at 50,000X if it is set up correctly, we do time after time.
I am still looking for a TEM for checking quality of the em products I produce. Are their any free or low-priced Zeiss 9s or similar instruments out there. I do not need particularly high resolution but rather a machine that is relatively easy to maintain. I would be most grateful for any help.
The corollary to this argument is that only items made by the supplier can be trusted. Frankly, I would worry about a supplier who makes WDS standards, grids, apertures, refilaments and more. Nothing wrong with an in-house facility to film grids, but what is wrong with the manufacturer maintaining standards? I know of three (not counting SPI) EM consumable suppliers who have their grids filmed by suppliers with TEMs. Presumably all such coated grids are made with TEM access and checking. This 'contribution' to the server by SPI is, all things considered, another blatant advertisement and a waste of subscribers time. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au
---------- } From: Garber, Charles A. {cgarber-at-2spi.com} } Vachik Hacopian wrote: } =================================================== } What is the best commercial source of consistently high-quality holey grids? } =================================================== } As a long time manufacturer of filmed grids, including "holey" and "lacey" } types, my answer would not be exactly that of an independent third party. } } However, just remember one thing: The "making" of filmed grids is easy to } describe, however the "art" of making a superb grid is not. But at the end } of the day, no one knows for sure what they have made unless they have their } own in-house TEM facilities to inspect the quality of their products. Be } certain your intended vendor has their own facilities to check themselves } what they are getting ready to send out their door. } } Otherwise you, the customer, will be the QC department for that vendor } yourself! Anyone who has been unhappy with purchased filmed grids in the } past will know exactly what I am talking about. Customers are always } welcome to visit our production facility and meet our staff members } responsible for the in-house production and TEM inspection of our coated } grids. } } Chuck } } =================================================== } Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 } President 1-(800)-2424-SPI } SPI SUPPLIES FAX: 1-(610)-436-5755 } PO BOX 656 e-mail: cgarber-at-2spi.com } West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com } } } Look for us! } ############################ } WWW: http://www.2spi.com } ############################ } ==================================================
We're looking for an apparatus for preparing vitrified TEM specimens, such as the Reichert KF80, which is not produced anymore. Does anybody have something like standing around?
Philip
-- Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 86 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www_scem.csb.ki.se/pages/philip.html
Dear colleagues, 35 mm.color slides are still a relevant media for us. We have been addressing the problem of PC/slide interface. We have obtained a slide scanner, so the problem of putting our large slide inventory on disk is solved. The slide printers we are aware of are too expensive (US$ 5,000 range?) I am considering setting up a good quality flat screen monitor with a 35 mm. camera and appropriate lens, dedicated to tranforming PC screens to slides. Does anyone have experience with such an arrangement, or a better solution? Thanks for your help, Prof. Walter A. Mannheimer Dept. of Metallurgy and Materiais Eng. Federal University of Rio de Janeiro POBox 68505, 21945 Rio de Janeiro, Brazil Vox (55 21) 590-0579 Fax (55 21) 290-6626 wamann-at-metalmat.ufrj.br
I have been making slides this way for years, it works very well. It works best with slow film (ASA 25 or 64) and long exposures (2 to 10 seconds) so that screen flicker is averaged out. I use a 120mm lens (longer lens yield less distortion), tripod, and cable release. Use as dark of room as you can to avoid glares on the screen. Also, taping matte black paper over the edges of the monitor prevents them from showing up on your slide, giving a much more polished look. Be sure to place the mouse pointer out of the way while taking the picture (I tend not to notice the mouse on the monitor as it looks natural there but on a slide it looks like you are trying to highlight a feature). If color balance is important for your slides then you may have to do quite a bit of fiddling with the monitor. Also, it is important to adjust the image width and height to remove distortions before taking pictures. The easy way to do this is to display an image you know is a circle and adjust the monitor until it really is a circle.
Dan Moore
At 09:38 AM 10/6/97 EST3EDT, you wrote: } Dear colleagues, } 35 mm.color slides are still a relevant media for us. } We have been addressing the problem of PC/slide interface. } We have obtained a slide scanner, so the problem of putting our large } slide inventory on disk is solved. } The slide printers we are aware of are too expensive (US$ 5,000 } range?) } I am considering setting up a good quality flat screen monitor with a } 35 mm. camera and appropriate lens, dedicated to tranforming PC screens } to slides. Does anyone have experience with such an arrangement, } or a better solution? Thanks for your help,
INTERNATIONAL CONFERENCE ON PHYSICAL MESOMECHANICS AND COMPUTER AIDED DESIGN OF ADVANCED MATERIALS AND TECHNOLOGIES - M E S O M E C H A N I C S ' 9 8 (May 31 - June 2, 1998)
and/or at
WORKSHOP ON MICRO- AND MESOMECHANICS ASPECTS OF MATERIAL FAILURE - M E S O F A I L U R E ' 98 (June 3 - 4, 1998)
in Tel Aviv, Israel (one paper per presenter).
What is Mesomechanics? A new science, a branch of the physics and mechanics of deformation and fracture, has recently received its own name - Mesomechanics. The "meso" range of experimental and theoretical analysis of processes related to deformation and fracture of solids is defined as lying between the scale at which continuum mechanics is sufficiently accurate for a description of the events, and the atomistic scale. Within this range, several subscales can be identified, for instance those related to grains, grain boundaries, particles, shear bands, voids, microcracks and dislocations. To study phenomena within the meso range, new tools are often needed, sometimes outside the normal use of classical mechanics. Mesomechanics deals also with phenomena of self-organization in materials under loading.
Conference and Workshop Co-Chairmen: Prof. V.E. Panin, Director of State Research Center Institute of Strength Physics and Materials Science, Siberian Branch of Russian Academy of Sciences, Tomsk, Russia. Prof. R.L. Salganik, Center for Technological Education Holon, Israel. Prof. G.C. Sih, Xi'an Jiaotong University, China. Prof. M.P. Wnuk, University of Wisconsin, Milwaukee, USA
Conference Topics: -Physics and mechanics of heterogeneous media as a basis for computer aided design of advanced materials. -Computer aided design of advanced materials based on metals, ceramics and polymers. -Basic scientific principles of strengthening and surface treatment of materials. -Non-destructive testing based on mesomechanics. -Mesomechanics of fatigue. -Experimental methods of mesomechanics. -Fractals in mesomechanics. -Mathematical models and methods for mesomechanics. -Contact mesomechanics. -Strain localization processes at pre-fracture stage on meso-level of material structure. -Influence of yielding, irreversible deformation and fracture on phase transformations in solids on micro- and meso-levels. -Mesomechanics of time-dependent deformation, damage and fracture. -High rate deformation and fracture processes. -Mesomechanics of highspeed and shock wave deformation. -Mesomechanics of rocks and soils -Engineering applications of mesomechanics.
Some invited key-note lectures that will be presented to the Conference: Prof. K.B. Broberg, "Modelling of Materials in Mesofracture" (University College Dublin, Ireland); Prof. Y.C. Gao,"Fatigue Mechanism of Fiber-Reinforced Materials" (Northern Jiaotong University, China); Prof. J.K. Knowles, "Continuum Modeling of Solid-Solid Phase Transitions" (Caltech, USA); Prof. B.R. Lawn, "Damage Accumulation Beneath Hertzian Contacts in Ceramics and Other Brittle Materials" (National Institute of Standards and Technology, USA); Prof. G.C. Sih, "Transitional and Stability Character of Mesofracture" (Xi'an Jiaotong University, China); Prof. M.P. Wnuk, "Constitutive Modeling of Damage Accumulation and Fracture in Multiphase Materials" (University of Wisconsin, Milwaukee, USA)
A synopsis of one page should be sent or faxed and also sent by e-mail (if possible) to the Conference and Workshop Coordinator not later than October 31, 1997: see the Application and Information Request below.
Please inform your Colleagues to submit tentative paper titles immediately, thanks.
For more information please see
http://www.cteh.ac.il/happen/meso98.html
or address:
Prof. R.L. Salganik - the Conference and the Workshop Coordinator. Center for Technological Education Holon P.O. Box 305, Holon 58102. Israel
****************************************** INTERNATIONAL CONFERENCE: "MESOMECHANICS'98" May 31. - June 2, 1998 WORKSHOP: "MESOFAILURE'98" June 3 - 4, 1998 Tel Aviv, Israel
APPLICATION AND INFORMATION REQUEST
Please mail this form to: Prof. R.L. Salganik - the Conference and Workshop Coordinator. Center for Technological Education Holon, Affiliated with Tel Aviv University P.O. Box 305, Holon 58102, Israel
I intend to: =C9 attend the Conference/Workshop (Yes/No), =C9 present an oral/poster paper to the Conference (Yes/No), =C9 present a paper to the Workshop (Yes/No) entitled:______________________________________________________ ___________________________________________________________ ___________________________________________________________ ___________________________________________________________ ___________________________________________________________ ___________________________________________________________
A synopsis of one page should be sent or faxed and also sent by e-mail (if possible) to the Conference and Workshop Coordinator not later than October 31, 1997.
=C9My organization intends to participate in the Exhibition (Yes/No).
} } Dear colleagues, } 35 mm.color slides are still a relevant media for us. } We have been addressing the problem of PC/slide interface. } We have obtained a slide scanner, so the problem of putting our large } slide inventory on disk is solved. } The slide printers we are aware of are too expensive (US$ 5,000 } range?) } I am considering setting up a good quality flat screen monitor with a } 35 mm. camera and appropriate lens, dedicated to tranforming PC screens } to slides. Does anyone have experience with such an arrangement, } or a better solution? Thanks for your help, } Prof. Walter A. Mannheimer } Dept. of Metallurgy and Materiais Eng. } Federal University of Rio de Janeiro } POBox 68505, 21945 Rio de Janeiro, Brazil } Vox (55 21) 590-0579 Fax (55 21) 290-6626 } wamann-at-metalmat.ufrj.br
Walter, I've been doing this for several years and am quite happy with the results. We have a slide making service here that accepts PC disks and charges too much money for slide production. With my tripod, macro lens and color slide film I can shoot and process an entire role of 36 exposures for less than $10US.
A good quality monitor is important. I meter directly off of the screen and use a mid range f stop 8 - 11. For text with a lot of light background, I bracket the exposure times. Turn off room lights to avoid reflections. I've compared slides from my monitor to those produced by a slide film recorder and yes, the resolution is better from the expensive dedicated slide film recorder. But not that much better.
My .02 worth.
cheers Ed
Edward J. Basgall, PhD The Pennsylvania State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618 http://www.personal.psu.edu/ejb11/
Has anyone performed image processing and analysis on nerve? Following is the question posed to me, I am clueless on what I would be measuring/doing regarding nerve analysis. I do not have any IA experience with nerves.
Input from Experts on neurology would be appreciated.
I suspect that there are probably reports of quantitative approaches to assessing peripheral nerves?? Does anyone have any suggestions?
Project:
I will be working with a diabetic drug, I believe designed to be an insulin "secretagog" (don't quote me on that) which in a previous mouse study caused a peripheral neuropathy morphologically similar to the classic spontaneous ageing neuropathy of mice. The hypothesis is that drug related changes in blood glucose contributed to the pathogenesis and thus it is a pharmacologic effect.
What questions should I be asking Gregory.Argentieri-at-pharma.novartis.com
At 09:00 AM 10/3/97 +0200, Jurgen Paetz wrote: } I would be interested to hear from other SEM users with the same (JOEL } JSM 5400) or a similar SEM what sort of filament life you are achieving? } I fear that we may have a vacuum leak since the filament life is } characteristically low and tarnishing is usually visible on the filament } holder. The latter I am told may be an indication of a vacuum leak in } the system.
I want to add my corroboration to this diagnosis. A combination of short filament life and discolored filament base is a sure indication of a vacuum leak or other source of contamination.
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
Hello to Prof. Mannheimer and those reading along,
I'm a technician and SEM operator at the Univ. of Penn. I've had very acceptable results in 'screen shooting' PC monitor images using a 35 mm Nikon camera and a slow color slide film. All we did was completely darken the room, (no lights and shaded the windows) to avoid any screen glare from these sources, put the camera on a good tripod with cable shutter release (exposures can be anywhere from 1/2 sec. to 5-10 sec. at full open lens); position the camera so that the screen image fills the field, adjust the screen brightness and contrast for an optimal appearing image (what you see is what you get) and shoot. We use the camera's interior light meter and usually bracket each shot. The bracketing can be reduced, or avoided, once you become comfortable with the film sensitivity you're using and the screen brightness of the image.
Also, Daniel Moore's procedures seem real good. Thanks for the tip on matte black paper.
Hello, I am hoping someone out there has had some experience doing in-situ techniques using semi-thin plastic sections. Frozen sections of the tissue I will be working with does not provide adequate morphology. Any suggestions or references would be greatly appreciated. Thanks in advance, Gary
Hello, I am hoping someone out there has had some experience doing in-situ techniques using semi-thin plastic sections. Frozen sections of the tissue I will be working with does not provide adequate morphology. Any suggestions or references would be greatly appreciated. Thanks in advance, Gary
wamann2-at-METALMAT.UFRJ.BR wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Dear colleagues, } 35 mm.color slides are still a relevant media for us. } We have been addressing the problem of PC/slide interface. } We have obtained a slide scanner, so the problem of putting our large } slide inventory on disk is solved. } The slide printers we are aware of are too expensive (US$ 5,000 } range?) } I am considering setting up a good quality flat screen monitor with a } 35 mm. camera and appropriate lens, dedicated to tranforming PC screens } to slides. Does anyone have experience with such an arrangement, } or a better solution? Thanks for your help, } Prof. Walter A. Mannheimer } Dept. of Metallurgy and Materiais Eng. } Federal University of Rio de Janeiro } POBox 68505, 21945 Rio de Janeiro, Brazil } Vox (55 21) 590-0579 Fax (55 21) 290-6626 } wamann-at-metalmat.ufrj.brDear Dr. Mannheimer,
I have taken a large number of slides in this fashion. I have an old Olynpus OS2N camera with a Vivitar 90 mm Macro/Telephoto lens on it. This sort of lens (prefferably with a zoom), gives you lots of flexibility. As I remember, we used tungsten film and set the shutter ot 1/60th second. Since we had a non-interlaced screen, this worked very well.
Hope this information is helpful. Best of luck!
Barbara Foster President Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108 PH: (413)746-6931 FX: (413)746-9311 email:mme-at-map.com --------------------------------------------------------------------------------------------------------------------------------- ********** Microscopy/Microscopy Education ********** America’s First National Consortium of Microscopy Experts Specializing in Customized, On-site Training in all areas of Microscopy, Sample Prep, and Image Analysis
Greg, Here's a publication that I was involved with years ago that did quantitation on peripheral nerve basement membrane thickness.
Johnson PC, Doll SC, Cromey DW (1986) Pathogenesis of diabetic neuropathy. Annals of Neurology 19:450-457.
Dr. Peter C. Johnson (formerly of the UA, and then the Barrow Neurological Institute in Phoenix AZ) and (as I recall) Dr. Peter J. Dyck (Mayo Clinic) were/are big users of morphometric approaches to peripheral nerves. I'd suggest running a MedLine on these two as a place to start for ideas.
Good Luck. Doug
At 05:11 PM 10/6/97 +0100, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America http://www.pharmacy.arizona.edu/exp_path.html Home of: "Microscopy and Imaging Resources on the WWW"
} How are the lines applied to TEM fluorescent screens? I have new screen } that is blank and would like to apply at least a recognizable spot at the } center, maybe a frame outlining the photo area. I'd think that I could use } a drafting pen. True? } AEI supplied us with a plastic template which had the proper posi- tions for the corners and centers marked for both "tilt" & "horiz" screen positions. We used to lay this over the screen and stick a probe through the holes in the plastic, then draw corners & crosses. Subsequently, we had our shop scribe the appropriate lines in our screen blanks with their smallest mill bit. When the phosphor is poured onto the blanks, the marks are quite visible. We also added marks for the middle of the film edges. Since there is ~2 mm uncertainty in the screen position from the mount, all these marks are only approximate. Yours, Bill Tivol
--IMA.Boundary.189261678 Content-Type: text/plain; charset=US-ASCII Content-Transfer-Encoding: 7bit Content-Description: cc:Mail note part
Owen,
Trying to remember how I marked screens that I had cast for the 1MeV microscope in Madison a long time ago. I think that I used an extremely sharp pointed, very soft lead pencil. As I recall, I sharpened the pencil on very fine sandpaper and the surface was hard enough to write on. I would expect that ink from a Rapidograph-type pen would wick and make a wide blotchy mark.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Good morning,
How are the lines applied to TEM fluorescent screens? I have new screen that is blank and would like to apply at least a recognizable spot at the center, maybe a frame outlining the photo area. I'd think that I could use a drafting pen. True?
TIA, Owen
Owen P. Mills Michigan Technological University Metallurgical & Materials Engineering Rm 512 MME Building Houghton, MI 49931 906-487-2002 906-487-2934 FAX opmills-at-mtu.edu --IMA.Boundary.189261678 Content-Type: text/plain; charset=US-ASCII; name="RFC822 message headers" Content-Transfer-Encoding: 7bit Content-Description: cc:Mail note part Content-Disposition: inline; filename="RFC822 message headers"
Received: from ns1 (159.198.180.56) by ccmailgw.mcgawpark.baxter.com with SMTP (IMA Internet Exchange 2.1 Enterprise) id 001E0E9F; Fri, 3 Oct 97 14:32:25 -0500 Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by ns1 (8.8.0/8.8.0) with SMTP id OAA20529; Fri, 3 Oct 1997 14:45:25 -0500 (CDT) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id KAA02536 for dist-Microscopy; Fri, 3 Oct 1997 10:39:03 -0500 Received: from news.mtu.edu (news.mtu.edu [141.219.70.11]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id KAA02533 for {Microscopy-at-Sparc5.Microscopy.Com} ; Fri, 3 Oct 1997 10:39:02 -0500 Received: from mtu.edu (root-at-mtu.edu [141.219.70.1]) by news.mtu.edu (8.8.7/8.8.7) with ESMTP id LAA12933 for {Microscopy-at-Sparc5.Microscopy.Com} ; Fri, 3 Oct 1997 11:48:44 -0400 (EDT) Received: from mmserver.mm.mtu.edu (mmserver.mm.mtu.edu [141.219.20.61]) by mtu.edu (8.8.7/8.8.7) with ESMTP id LAA05906 for {Microscopy-at-Sparc5.Microscopy.Com} ; Fri, 3 Oct 1997 11:48:43 -0400 (EDT) Received: from Backscatter.my.mtu.edu (backscatter.my.mtu.edu [141.219.20.99]) by mmserver.mm.mtu.edu (8.8.7/8.8.7/mturelay-1.2) with SMTP id LAA18203 for {Microscopy-at-Sparc5.Microscopy.Com} ; Fri, 3 Oct 1997 11:48:42 -0400 (EDT) Message-Id: {3.0.32.19971003114147.006d2cbc-at-mmserver.mm.mtu.edu} X-Sender: opmills-at-mmserver.mm.mtu.edu X-Mailer: Windows Eudora Pro Version 3.0 (32)
REDUCED FEE REGISTRATION DEADLINE January 31, 1998 Workshop on Tripod Polishing
Workshop Objective This course will cover all aspects of pre-thinning and focus on final thinning for TEM via Tripod Polishing. Due to the limited class size an= d the extensive hands-on opportuinities, this course is well suited to novices as well as advanced Tripodders. Attendees will also learn the latest techniques available in ion milling and in plasma cleaning for TEM=
samples. The course will include sections on:
How to do it and why should I? What's really going on and what am I really seeing? How to prepare small, specific area cross-sections. The problem of wildly differing materials (eg tungsten). Rapid preparation of TEM cross-sections. Preparation of a wide range of materials: semiconductors, ceramics, metals,...
Hands-on Opportunity This course will be unique in that it will provide a hands-on opportunity=
for every class participant. Tripod Polishers, Polishing Wheels, and pre-thinning equipment will be made available to all participants and actual samples will be prepared - by the students - as part of the course= =2E =
This is a great opportunity to get your hands dirty and actually learn by=
doing. The instructors will walk you through each step of the process an= d then let you loose on the equipment. This course is designed to teach th= e Tripod Polishing technique. Silicon samples will be provided to the students and used as the basis for the course teaching.
Workshop Location and Dates South Bay Technology - San Clemente, CA Dates: Friday & Saturday - March 13 & 14
Previous Participants (partial list) INTEL, AMD, Motorola, LSI Logic, Conner Peripherals, Univ of Maryland, Un= iv of New Mexico, UNAM (Mexico), LG Electronics (Korea), Battelle, MEMC, MVA=
Inc., Univ of Michigan, U.S. Bureau of Mines, IBM, Naval Research Lab, Purdue Univ, Univ of Alabama, Univ of Arizona, Univ of Colorado, Univ of Wisconsin.
Class Size Due to the intensive hands-on aspects of this course, class size will be strictly limited to 10 participants.
Registration Fee: $795 (includes lunches and Friday night Dinner) $695 if registration fee paid by January 31, 1998=
Registration Deadline: 30 days prior to workshop
For additional Information: Monica Pflaster South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 TEL: 800-728-2233 FAX: 714-492-1499 e-mail: sbt-at-southbaytech.com
ON-LINE Registration available at: http://www.southbaytech.com
Registration Form
To register for the workshop, please fill out this form and send it, with=
registration fee to:
South Bay Technology, Inc. =
Workshop on Tripod Polishing 1120 Via Callejon San Clemente, CA 92673 USA
Payment must be made in the form of a check, money order, Visa or MasterCard. Checks must be drawn on a U.S. Bank and made payable to Sout= h Bay Technology, Inc. Credit card orders by FAX may be sent to South Bay Technology at 714-492-1499. Please do not send credit card information v= ia e-mail.
Name: = =
=
Affiliation: = =
=
Address: = =
=
= =
=
=
City: State: = =
Zip: Country:_________ Telephone: FAX: = =
=
e-mail:________________________
Primary sample type: = =
= =
=
VISA MasterCard Card #_________________________________
Expiration Date________ Signature of Cardholder_________________________
Cardholder name (Please print):________________________________________ =
We are embarking on some bromo-deoxyUridine (BrDU) studies with cell cultures and are wondering if we need to make up fresh stocks each time or can we sterile filter and store at 4 C or -80C. Does anybody have practical experience with its stability? TIA.
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
I know this is sort of off the topic of the listserv, but can anyone help a colleague who is:
looking for any information on a company or individual who sells Anatoxin-a(s) [P=O structure]. This is a toxin isolated from the blue-green freshwater algae Anabaena flos-aquae.
Or if anyone knows how the toxicity differs between anatoxin-a(s) and the synthetic anatoxin-a fumarate (produced by Sigma and others).
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Jurgen Paetz wrote: } } } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } -----------------------------------------------------------------------.} } } Dear fellow SEM users } } } } I would be interested to hear from other SEM users with the same (JOEL } } JSM 5400) or a similar SEM what sort of filament life you are achieving } } ? } } } } I fear that we may have a vacuum leak since the filament life is } } characteristically low and tarnishing is usually visible on the filament } } holder. The latter I am told may be an indication of a vacuum leak in } } the system. } } } } Regards } } } } J. Paetz (Senior mineralogist) } } } } Amplats Research Center } } Republic of South Africa } } } } Dear Jurgen Paetz, } } You are correct that a discolored base is usually a sign of a poor } vacuum. A normal burnout of a filament should have a bubble on top of } the broken wire. If your filament burns out this way and the base is } discolored it is probably a bad vacuum. If the filament is craked , no } bubble, then their could be a flaw in the wire or a slight crack was } made by human handling. } We do not use a JEOL ourselves, but we do sell filaments for all the } different scopes and our customers seem to feel you should get 50 - 200 } hours out of a filament. } } John Arnott } Ladd Research } We have a JEOL 5800LV. Our filiment life on that machine has been 150-200. We feel that is low. Our JEOL 733 filiments last in the 1000's of hours. The 5800 burnt filiments display the small "ball" (normally only present on one end of the break unless the filiment has been very over-driven). The bases of the filiments are always slightly discolored and we do not have a vacuum leak (that we know of). The bases may get discolored when we work in LV mode (though I have been assured that the vacuum in the gun chamber stays very high). Our JEOL 733 filiment bases are always somewhat discolored, and we watch the gun chamber vacuum very closly, so I know there is no leak.
One thing that may extend the filiment life is allowing the filiment to cool before venting the chamber (I don't know if the 5400 uses an exchange port or just vents the chamber). On our Hitachi S-450 we found that doing this does increase filiment life, but the 5800 is new and we have just started cooling the filiment, so I do not know what the effect will be. I do know that machines that change Acc kV on the fly have shorter filiment lives.
I write this not to contradict John Arnott's above statement, but just to show that there are "extreme" differences in machine filiment lives that are not just based on the type of machine being used but also possibly on the way the machine is used. The number 50-200 hours seems low to me, but most of my experience is on the 733, so maybe I am spoiled. As far as discoloration being a sign of a leak, I bet it is, but how much discoloration is normal should also be a question.
We are doing TEM on cultured Koala lymphocytes. We are having an on going problem with Spurr's resin (we use medium Spurr's). In some cases resin doesn't get polymerised and stays soft even after 3-4 days in sixty degree oven. The interesting point is that it doesn't happen with every sample. Even in a series of different samples which are processed at the same time and embeded in the same batch of resin, some remain soft while the others are quite Ok.
We heard that some components of culture media may interfere with resin polymerisation so we have been careful to wash the cultured cells properly but it did not eliminate the problem. Any advice on how to tackle the problem is highly appreciated. I would also be thankful if you suggest a way to revive those samples which are embeded in soft resin.
Regards
M. Ghoddusi
Majid Ghoddusi Division of Veterinary Pathobiology The University of Queensalnd QLD 4072 Australia
My colleague Neil Hand has recently published a paper entitled:
Non-isotopic in-situ hybridization to detect chick Sox gene mRNA in plastic embedded tissue. Histochemical Journal Vol 29, pp 625-629 (1997)
This may be of help, including other references.
Neil may be contacted at: mpznhand-at-unix.ccc.nottingham.ac.uk
Cheers
Kev
^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ Kevin J. Randall Dept of Histopathology Queen's Medical Centre University Hospital NHS Trust Nottingham NG7 2UH Tel: 0115 924 9924 x 43725 ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^K
We need to analise Cu, Ag, Fe, etc. as impurities in very small samples of gold alloys. The best technique that we could find is thin foild EDS using an analytical Hitachi 8100 TEM with a Ge detector. To improve accuracy, we would like to use elemental standards, but we did not find a supplier of EDS/TEM standards up to now. I would appreciate to receive your suggestions.
Best regards Rui Vilar DeMAT/IST -- ####################################### Rui Vilar Departamento de Engenharia de Materiais Instituto Superior Ticnico Av. Rovisco Pais, 1096 Lisboa Codex Portugal Tel.: -351-1-8418121 Fax: -351-1-8418121 or -351-1-8418120 Email: pcrvilar-at-alfa.ist.utl.pt #######################################
reidr1-at-uofs.edu wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Email: reidr1-at-uofs.edu } Name: Richard Reid } } School: University of Scranton } } Hello, } I am an undergrad attending the University } of Scranton. I am trying to find some beginner } information regarding flourescent microscopy. } While I have found some web-sites dealing with } flourescence, they are all too advanced for me. } It is the same situation with the school's } library. Most of the information is not geared } for the beginner. I am mostly interested in } staining neural tissue (CNS). } } Thank you very much for any help you can } offer. It will be greatly appreciated. } } Sincerely, } } Richard Reid } } --------------------------------------------------------------------------- Dear Richard,
We occasionally run a class in fluorescence. The best resource for the class has been a set of books by ROST: Fluorescence Microscopy (Vol 1), and Quantitative Fluorescence Microscopy. Publisher: Cambridge Press.
Our new book "Optimizing Light Microscopy for Biological and Clinical Labs" also has a sound chapter on Fluorescence. Email me for details, if you are interested.
Hope these help. Good luck in your studies.
Best regards,
Barbara Foster President Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108 PH: (413)746-6931 FX: (413)746-9311 email:mme-at-map.com --------------------------------------------------------------------------------------------------------------------------------- ********** Microscopy/Microscopy Education ********** America’s First National Consortium of Microscopy Experts Specializing in Customized, On-site Training in all areas of Microscopy, Sample Prep, and Image Analysis
Fellow micrsocpist, Has anyone had experience identifying blood residue on obsidian stone tools using sem. These tools are between 800 to 1000 years old. Does anyone know of any references that bases identification of blood on morphology or elemental analysis or has any hints for features that may be apparent?
Hank Adams Electron Microscopy Lab New Mexico State University, :"Home of the worst football team in the country" and proud of it!
EMPLOYMENT ANNOUNCEMENT: ElectroImage has an immediate opening for a technical person who has microscopy, computer, and image analysis skills. We are based on Long Island and distribute digital cameras, software, and other imaging products. If you would be interested in discussing the opportunity, please contact me at Matt-at-electroimage.com or by telephone at 516-773-4305.
Thank you.
Matt Irwin ElectroImage, Inc. 277 Northern Blvd. Suite 101 Great Neck, NY 11021
First of all, please DO NOT "REPLY" to this message, but contact the person listed below, for whom I am posting this message to Microscopy (she is not subscribed). Thanks. Gib Ahlstrand.
Used JEOL 100B TEM, free giveaway, but you pay shipping costs. This scope is in good working order with lots of extras: EDAX brand x-ray microanalaysis system (older model), SED detector, STEM detector, specimen current detector.
Contact: Ms. Barb Clark, University of Minnesota, Dept. of OBGYN, Minneapolis, MN USA 55455.
One of the companies would be M.A.C. in the U.K. E-mail standards-at-dial.pipex.com Fax: +44 1480 462901 they also have a web page i think. htpp://www.macstandards.co.uk/town/street/yr49/ ------------------------------------- Name: Luc Harmsen E-mail: anaspec-at-.icon.co.za
Vachik Hacopian wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } What is the best commercial source of consistently high-quality holey grids? } } Vachik Hacopian
Dr. Hacopian, I am inclined to say that since Ladd has been producing holey film for over forty years that we produce the most consistent and best film , but I suppose all manufacturers would say the same thing or go into a lomg-winded discourse on why theirs is the best. I would suggest that the best course for you is to try some different suppliers who have a long history of supplying high quality products over the years (such as Ladd, Pella Fullam, etc.) and see whose film best suits your application. Since their reputations for quality have existed for such a long period I don't think you would go wrong with any of them.
Developmental Biologist Department of Zoology Miami University Oxford, Ohio
We seek an Assistant Professor for a tenure-track position to begin in August, 1998. Ph.D. in zoology or biology and postdoctoral experience required. Individuals with expertise in any area of Animal Developmental Biology are invited to apply, but preference will be given to those who use electron microscopy as a major research tool. We expect this person to develop an independent research program in developmental biology that will enhance the department's research capabilities.
Teaching responsibilities include: (1) a sophomore level course in developmental biology; (2) participation in a team-taught introductory biology course; and (3) and advanced course in a specialty area. The successful applicant will be expected to seek external funding to support his/her research and to supervise and advise graduate and undergraduate students. Advancement will be based on teaching, research, and professional service, with primary emphasis on teaching and research.
Miami University is a state-assisted institution in SW Ohio. The department has excellent research facilities; the EM facility is well-equipped for SEM, TEM, cryopreservation, and confocal microscopy (see http://www.muohio.edu/~zoocswis for more details about the department and our facilities). The department has strong Ph.D. and M.S. programs, 32 faculty members, several postdoctoral researchers, and 50 graduate students on the Oxford campus.
Interested persons should submit a curriculum vitae, a statement of teaching philosophy, a description of current research and long-term research interests, and should arrange for three letters of recommendation and transcripts of graduate and undergraduate academic work to be sent to:
Dr. Douglas H. Taylor, Chair of Zoology, Miami University, Oxford, OH 45056.
Review of applications will begin on 1 December, 1997, and continue until the position is filled. Miami University offers equal opportunity in employment and education.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"640K ought to be enough for anybody." -- Bill Gates, 1981
I use a 160 tube length upright Zeiss microscope for plankton research.Because I have to study living cells I want to use DIC (differential interference contrast or Nomarkski). I asked Zeiss in the UK and in the Netherlands , but they told me that this device is not available anymore for that kind of "old" type microscopes. Knows anybody an address where I can get secondhand DIC attachment.
"Filament Life" The life of a filament in an electron probe has been mentioned during the=
discussions of SEM filament life, they cannot be compared!
A filament in a probe is being used to generate x-rays not to produce a high resolution image, the cap design and component geometry is different=
from a high resolution SEM. Those of you who use your SEM for EDX work will know that with the correct geometry you do not need much beam curren= t to generate a good x-ray spectra; people use lower emission currents or much smaller spot sizes to compensate for the SEM overcurrent.
"Filament Base" Filament bases indicate the environment in which that filament is being forced to operate. A good vacuum and economy filament position (TEM styl= e) even in an SEM will give a base lightly coated with tungsten, which is powder blue in colour. Drive the filament harder and the base will gathe= r even more tungsten and become much darker. If the vacuum environment is = of a poor quality the filament base will become yellow to orange in colour, this IS an indication of a poor environment which, in spite of what the vacuum gauges say, indicates a leak or an inefficient vacuum system. =
"Virtual Leaks" Be aware that the practice of situating the diffusion pump or turbo pump=
immediately below the specimen chamber does mean that filament life is related to how dirty the specimen is. A gassy specimen will reduce the pumping efficiency of the gun and thus filament life. A bad vacuum in th= e gun produces discharge and thus instability but most of all you will smel= l the problem when you open the gun; it has an oily-ozone smell!
Thanks to all who responded to my question. Quite a number of people photograph a monitor with good results. The several hints are appreciated. Best regards to all Prof. Walter A. Mannheimer Dept. of Metallurgy and Materiais Eng. Federal University of Rio de Janeiro POBox 68505, 21945 Rio de Janeiro, Brazil Vox (55 21) 590-0579 Fax (55 21) 290-6626 wamann-at-metalmat.ufrj.br
We are currently observing gold foils in TEM, along with EDS analysis. However, in order to perform absortion corrections we need to know the thickness of the sample. Question: how can we determine sample thickness from TEM observations? (We have a double tilt analytical holder).
} } Dear All } } We are doing TEM on cultured Koala lymphocytes. We are having an on going } problem with Spurr's resin (we use medium Spurr's). In some cases resin } doesn't get polymerised and stays soft even after 3-4 days in sixty degree } oven. The interesting point is that it doesn't happen with every sample. } Even in a series of different samples which are processed at the same } time and embeded in the same batch of resin, some remain soft while the } others are quite Ok. } } We heard that some components of culture media may interfere with resin } polymerisation so we have been careful to wash the cultured cells properly } but it did not eliminate the problem. Any advice on how to tackle the } problem is highly appreciated. I would also be thankful if you suggest a } way to revive those samples which are embeded in soft resin. } } Regards } } M. Ghoddusi } } Majid Ghoddusi
Dear Majid Ghoddusi,
Please keep in mind that Ladd is a supplier of all the chemicals dicussed but with that in mind I would like to suggest the following:
1) It is very important to thoroughly mix the complete resin mixture. From your brief description, incomplete mixing would seem a likely reason for your problems. 2) If you are sure that incomplete mixing is not the problem, have you allowed enough time for complete infiltration of complete resin into your cells? 3) Humidity might also be an issue. The resin mixture is hygroscopic thus the resin may be absorbing water which would adversly effect curing and cutting properties. Try curing in a closed BEEM or gelatin capsule.
A solution that sometimes extracts epoxy reins from embedded samples can be prepared as follows: a) Prepare standard solution of KOH in absolute ethanol b) Allow to stand overnight c) The dark-colored supernatant is used as solvent d) Trim areas that contain only epoxy resin and immerse the sample in the solvent e) After epoxy resin is disolved wash 4 or so times in absolute ethanol F) Re-embed CAUTION!!!!!! I can not predict how if this treatment will destroy the vital components of your cells so please test this procedure or a couple of samples.
} } We need to analise Cu, Ag, Fe, etc. as impurities in very small samples } of gold alloys. The best technique that we could find is thin foild EDS } using an analytical Hitachi 8100 TEM with a Ge detector. To improve } accuracy, we would like to use elemental standards, but we did not find } a supplier of EDS/TEM standards up to now. I would appreciate to receive } your suggestions. } } Best regards } Rui Vilar } DeMAT/IST } -- } ####################################### } Rui Vilar } Departamento de Engenharia de Materiais } Instituto Superior Ticnico } Av. Rovisco Pais, 1096 Lisboa Codex } Portugal } Tel.: -351-1-8418121 } Fax: -351-1-8418121 or -351-1-8418120 } Email: pcrvilar-at-alfa.ist.utl.pt } #######################################
Dear Rui Vilar: Because thickness of specimen and standards are very difficult to control within 10 or even 20% of desired, quantitative analysis in TEM is elusive. Thickness (and other factors) change X-ray generation greatly. There are always exceptions, for instance just comparing a simple (say 2 phase) spectrum with that of a very similar standard will give reasonable results.
If the impurities are smaller than about 4 microns the area analysed will be too small for an EDS on SEM and the smaller 'envelope' analysed in TEM 'wins'. But TEM analysis could never be 'quantitative' - e.g. +/- 1% in a hundred.
In TEM, beam diameter (plus a little) yields most X-rays, but the small analysis envelope is little help if the impurities are not uniform throughout the thickness of the specimen as well.
Few companies make TEM EDS/WDS standards because of their limited applications. Regards Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au
} -----------------------------------------------------------------------. } } We need to analise Cu, Ag, Fe, etc. as impurities in very small samples } of gold alloys. The best technique that we could find is thin foild EDS } using an analytical Hitachi 8100 TEM with a Ge detector. To improve } accuracy, we would like to use elemental standards, but we did not find } a supplier of EDS/TEM standards up to now. I would appreciate to receive } your suggestions. } } Best regards } Rui Vilar } DeMAT/IST } -- } ####################################### } Rui Vilar } Departamento de Engenharia de Materiais } Instituto Superior Ticnico } Av. Rovisco Pais, 1096 Lisboa Codex } Portugal } Tel.: -351-1-8418121 } Fax: -351-1-8418121 or -351-1-8418120 } Email: pcrvilar-at-alfa.ist.utl.pt } #######################################
Our facility is undergoing ISO certification. My EM lab is small, consisting of one SEM with BSE and EDS. I would appreciate any suggestions/warnings, etc. from your experiences with ISO certification.
You can use convergent beam diffraction to measure thickness. In principle, this relies on the measurement of the spacings of thickness fringes which are visible in the CBED disks. It is very well described in the book "Transmission Electron Microscopy", volume II, "Diffraction" by D. B. Williams and C. B. Carter (Plenum, 1996). For the development of the technique they quote works by P. M. Kelly et al, phys. stat. sol. A31 (1975) p. 771, and S. M. Allen, Phil. Mag. A43 (1981) p. 325.
This is probably the best bet, and may be fairly straightforward for gold foils.
Wharton Sinkler
On Wed, 8 Oct 1997, Nuno Braz wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } We are currently observing gold foils in TEM, along with EDS analysis. } However, in order to perform absortion corrections we need to know the } thickness of the sample. } Question: how can we determine sample thickness from TEM observations? } (We have a double tilt analytical holder). } } Email: pcrvilar-at-alfa.ist.utl.pt } Tel: 351 1 8418124 } Fax: 351 1 8418120 }
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Wharton Sinkler PhD Department of Materials Science and Engineering Northwestern University 2225 North Campus Drive Evanston, IL 60208-3108 tel: (847) 491-7809 fax: (847) 491-7820 email: sinkler-at-apollo.numis.nwu.edu
Now that the subject of holey-C grids has been raised, are there any vendors willing to guarantee contamination-free films? Most of the grids I've purchased from vendors will contain some silicon contamination, along with variable Na, Ca, K and Cl. Since I normally investigate silicates, Si contamination is very problematic. I assume the Si comes from diffusion pump oil. Have any of the vendors with "TEM quality control" checked for the cleanliness of their grids?
Ciao for now, Ken
Kenneth JT Livi Department of Earth and Planetary Sciences 34th and Charles Streets Johns Hopkins University Baltimore, Maryland 21218 USA Phone: (410) 516-8342 Fax: (410) 516-7933 e-mail: klivi-at-jhu.edu
--IMA.Boundary.048223678 Content-Type: text/plain; charset=US-ASCII Content-Transfer-Encoding: 7bit Content-Description: cc:Mail note part
Hank,
It just so happens... See: Material Residues on Stone Tool Edges: Is Optical Microscopy Missing an Opportunity. Microscope Vol 45:3 89-93 (1997). Just out. On first page is topic heading BLOOD TRACES. Let me know if you need a fax.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Fellow micrsocpist, Has anyone had experience identifying blood residue on obsidian stone tools using sem. These tools are between 800 to 1000 years old. Does anyone know of any references that bases identification of blood on morphology or elemental analysis or has any hints for features that may be apparent?
Hank Adams Electron Microscopy Lab New Mexico State University, :"Home of the worst football team in the country" and proud of it! --IMA.Boundary.048223678 Content-Type: text/plain; charset=US-ASCII; name="RFC822 message headers" Content-Transfer-Encoding: 7bit Content-Description: cc:Mail note part Content-Disposition: inline; filename="RFC822 message headers"
Received: from ns2.baxter.com (159.198.1.38) by ccmailgw.mcgawpark.baxter.com
(IMA Internet Exchange 2.1 Enterprise) id 001E7B96; Tue, 7 Oct 97 15:55:48 -0500 Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by ns2.baxter.com (8.8.0/8.8.0) with SMTP id QAA28048; Tue, 7 Oct 1997 16:04:37 -0500 (CDT) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id MAA11625 for dist-Microscopy; Tue, 7 Oct 1997 12:03:33 -0500 Received: from bubba.NMSU.Edu (bubba.NMSU.Edu [128.123.3.39]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id MAA11622 for {Microscopy-at-Sparc5.Microscopy.Com} ; Tue, 7 Oct 1997 12:03:26 -0500 Received: from NMSU.Edu by bubba.NMSU.Edu (8.8.7/NMSU) id LAA25942; Tue, 7 Oct 1997 11:16:33 -0600 (MDT) id LAA06558; Tue, 7 Oct 1997 11:12:49 -0600 (MDT) Received: from Microspace.nmsu.edu (pc-hadams.NMSU.Edu [128.123.5.46]) by nestor.NMSU.Edu (8.8.6/8.7) with SMTP id LAA100884 for {Microscopy-at-Sparc5.Microscopy.Com} ; Tue, 7 Oct 1997 11:13:23 -0600 Message-Id: {199710071713.LAA100884-at-nestor.NMSU.Edu} Comments: Authenticated sender is {hadams-at-cnmailsvr.nmsu.edu} To: Microscopy-at-sparc5.microscopy.com
There are several methods you can use for this. Depending on the nature of your sample , some of these might be more " practical " than other methods.
1) You might be able to see a contamination spot on the sample. If so, then if you tilt the sample a known amount you can get a value of the thickness by measuring the distance between the contamination spots at the top and bottom of the foil.
2) You might be able to use convergent beam analysis. (see for example D.B. Williams Practical Analytical Electron Microscopy in Materials Science).
3) You can estimate the foil thickness from the number of thickness fringes seen using two beam conditions ( see for example Edington's book " Practical Electron Microscopy in Materials Science" ).
4) You can also use trace methods (see Hirsch et. al).
Jordi Marti
We are currently observing gold foils in TEM, along with EDS analysis. However, in order to perform absortion corrections we need to know the thickness of the sample. Question: how can we determine sample thickness from TEM observations? (We have a double tilt analytical holder).
} Mounting medium can add tens of } microns (or hundreds, if you're having a bad day!) to the effective } thickness of } the coverslip. Along with variability in thickness of coverslips (--even } those } that are nominally 0.17 mm--), that's the reason for the addition of } correction } collars onto lenses. } One way around that problem is to mount sections directly on the coverslip surface and then use the mounting medium to attach the coverslip the slide. With this approach, there is no chance of having to much mounting medium between the coverslip and specimen. But as Barbara Foster pointed out in another posting, we find great variation in actual coverslip thickness between manufacturers and within a box. We have also found supposedly high quality glass slides which varied significantly in thickness along the length of an individual slide. this can also lead to excessive mounting medium between the coverslip and slide if the coverslip spans a low point in the center of the slide. An inexpensive micrometer is the only way you can be sure you are buying a good product.
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. Steve Chapman, That was so well put that I printed it into our operating manual. It still leaves me with the questions, "Is 150-200 hours all I can expect from our JEOL 5800LV filaments?, and should we expect less filament life if we run at low vacuum more often?" I have been told "no" to the second question, but I believe that may be wrong. As far as the first question goes, I have been working with probes so long that 200 hours just seems very low (even keeping in mind that SEM filaments work far harder). Does cooling the filament before venting the chamber extend filament life?? Filaments only cost ~$80 each, but we are soft funded, so saving a couple hundred over a year actually helps us alot. Thanx Christopher
} } "Filament Life" } The life of a filament in an electron probe has been mentioned during the } discussions of SEM filament life, they cannot be compared! } } A filament in a probe is being used to generate x-rays not to produce a } high resolution image, the cap design and component geometry is different } from a high resolution SEM. Those of you who use your SEM for EDX work } will know that with the correct geometry you do not need much beam current } to generate a good x-ray spectra; people use lower emission currents or } much smaller spot sizes to compensate for the SEM overcurrent. } } "Filament Base" } Filament bases indicate the environment in which that filament is being } forced to operate. A good vacuum and economy filament position (TEM style) } even in an SEM will give a base lightly coated with tungsten, which is } powder blue in colour. Drive the filament harder and the base will gather } even more tungsten and become much darker. If the vacuum environment is of } a poor quality the filament base will become yellow to orange in colour, } this IS an indication of a poor environment which, in spite of what the } vacuum gauges say, indicates a leak or an inefficient vacuum system. } } "Virtual Leaks" } Be aware that the practice of situating the diffusion pump or turbo pump } immediately below the specimen chamber does mean that filament life is } related to how dirty the specimen is. A gassy specimen will reduce the } pumping efficiency of the gun and thus filament life. A bad vacuum in the } gun produces discharge and thus instability but most of all you will smell } the problem when you open the gun; it has an oily-ozone smell! } } Steve Chapman } Senior Consultant } Protrain }
We have had good success in many cases using the Bremmstrahlung shape to determine an absorption correction. See "EMAG '87 - Anlytical Electron Microscopy", ed. G. W. Lorimer, Institute of Metals, London (1988) p. 7, or "Analytical Electron Microscopy 1987" ed. D. C. Joy, San Francisco Press, p. 225 for more information. In the work we reported there we used software we wrote ourselves. We now use "Desktop Spectrum Analyser" from the folks at NIH. I don't know if any of the commercial analyser software packages support this algorithm.
We have never, so far as I remember, used this method in gold, but I don't see why there should be any problem.
This method avaoids having to determine the sample thickness (notoriously difficult!) or having to make any assumptions about the sample geometry.
I'd be glad to provide more information if you e-mail me directly.
Tony Garratt-Reed
Anthony J. Garratt-Reed MIT Room 13-1027 77 Massachusetts Avenue Cambridge, MA 02139-4307 United States of America
I am up against a brick wall in the calibration of a rotating compensator. For our ISO program, we must have all measurement devices calibrated by a traceable (NIST) standard if possible. Are there any ideas on how to calibrate a rotating compensator for birefringence measurements.
Jim Harper Amoco Fabrics and Fibers jeharper-at-amoco.com
The easy (or easiest) and accurate method to measure the thickness of the specimen with a known structure like gold is Two-beam CBED. From the rocking curves you see from the CBED pattern you can measure the thickness. If you use a Philips CM microscope, there is a freeware to measure the thickness from the CBED pattern. Although I never use that software by myself, I believe it is quite straigh forward. Or you can do a much more accurat thickness refinement by using J.M.Zou's refinement program or other similar program. Check out J.C.H.Spence's book.
Yifan
-- ********************************************************************** * Dr. Yifan Cheng * Phone: +1-850-644-4104 (Office)* * Institute of Molecular Biophysics * or: +1-850-644-9769 (Lab) * * Florida State University * Fax: +1-850-561-1406 * * Tallahassee, FL 32306-4380 * Email: ycheng-at-sb.fsu.edu * * U.S.A. * http://www.sb.fsu.edu/~ycheng * **********************************************************************
Sorry, but we had a typo in my previous e-mail. Part A should have read Prepare a "saturated" solution not a "standard" solution. Hope this is clear. If not, please e-mail or call 802-878-6711 or fax at 802-878-8074.
Sounds like an infiltration problem. What is your procedure? Are your blocks too big? Is your infiltration time too short? Is your dehydrating agent wet? Are your components well mixed before use? Do you let the first 3 components mix well before adding the catalyst? Have you at any time let the surface of the sample dry so that further infiltration doesn't happen properly?
On Tue, 7 Oct 1997, Majid Ghoddusi wrote:
} Date: Tue, 7 Oct 1997 09:21:55 +1000 (GMT+1000) } From: Majid Ghoddusi {vp092327-at-student.uq.edu.au} } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM: Problem with Spurr's resin } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Dear All } } We are doing TEM on cultured Koala lymphocytes. We are having an on going } problem with Spurr's resin (we use medium Spurr's). In some cases resin } doesn't get polymerised and stays soft even after 3-4 days in sixty degree } oven. The interesting point is that it doesn't happen with every sample. } Even in a series of different samples which are processed at the same } time and embeded in the same batch of resin, some remain soft while the } others are quite Ok. } } We heard that some components of culture media may interfere with resin } polymerisation so we have been careful to wash the cultured cells properly } but it did not eliminate the problem. Any advice on how to tackle the } problem is highly appreciated. I would also be thankful if you suggest a } way to revive those samples which are embeded in soft resin. } } Regards } } M. Ghoddusi } } } Majid Ghoddusi } Division of Veterinary Pathobiology } The University of Queensalnd } QLD 4072 } Australia } } Tel: (07) 3365 2569 } Fax: (07) 3365 1355 }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Dear Nuno: Another way you can use to measure thickness, if you have the Gatan EL/P system, is EELS. By measuring the intensity of Zero loss and the first plasma peak, you can figure out the thickness. This method is described on Egerton's "EELS in Electron Microscopy" book. Good luck!
On Wed, 8 Oct 1997, Nuno Braz wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } We are currently observing gold foils in TEM, along with EDS analysis. } However, in order to perform absortion corrections we need to know the } thickness of the sample. } Question: how can we determine sample thickness from TEM observations? } (We have a double tilt analytical holder). } } Email: pcrvilar-at-alfa.ist.utl.pt } Tel: 351 1 8418124 } Fax: 351 1 8418120 }
} . . . } } "Filament Life" } The life of a filament in an electron probe has been mentioned during the } discussions of SEM filament life, they cannot be compared! } } A filament in a probe is being used to generate x-rays not to produce a } high resolution image, the cap design and component geometry is different } from a high resolution SEM. Those of you who use your SEM for EDX work } will know that with the correct geometry you do not need much beam current } to generate a good x-ray spectra; people use lower emission currents or } much smaller spot sizes to compensate for the SEM overcurrent. } } . . .
I agree that microprobes are designed quite differently than SEMs ... but I am not too sure to what extent this can be said of their respective guns. While I can well imagine a probe gun having design aspects for stability (... a cross-over less susceptable to mechanical or thermal variation ...), resultant beam currents for x-rays (edx or wdx) vs. imaging are a result of condenser lens settings. That is, both of my instruments (probe and SEM) use very similar emission currents (i.e., 60 to 80 microAmps) ... measured as the load on the HV power supply. While tungsten filament saturations for either type of gun do involve filament tip design and wehnelt geometries, I am willing to bet saturation (self-biasing as a result of "driving the filament heat") occurs within 25 degrees of eachother, and that there probably is as great a disparity between SEM manufacturers than SEM vs. probes. Of course, you should feel free to clarify what you mean by choosing lower "emission" currents and ""SEM overcurrent" ...
cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
Spaulding, Robert F wrote: } } I am looking for a source for the book, Quantitative Electron-Probe } Microanalysis by V. D. Scott and G. Love. My local bookstore is having } no luck. } } Thanks, } Robert F. Spaulding } Corning Incorporated } Characterization Science & Services } Microscopy & Microanalysis Dept. } SP-FR-01-8 } Corning, New York 14831 } } 607-974-3732 } fax 607-974-3385 } Internet: SpauldinRF-at-corning.com
Publisher is Ellis Horwood (New York, London, Toronto), ISBN 0-13-104050-2 and address is:
Ellis Horwood Limited A division of Simon & Schuster International Group Campus 400, Maylands Avenue Hemel Hempstead Hertfordshire, HP2 7EZ UK
Henrik -- Henrik Kaker SEM-EDS Laboratory Metal Ravne d.o.o. Koroska c. 14 2390 Ravne Slovenia Tel: +386-602-21-131 Fax: +386-602-20-436 SEM-EDS Lab http://www2.arnes.si/guest/sgszmera1/index.html MVD Database http://www.kaker.com/mvd/vendors.html Kaker.Com http://www.kaker.com
Although my primary responsibility is running our EM facility, I have served as an R&D team representative in obtaining ISO 9001 certification, and I have also been trained in auditing. Hopefully, I can give you some appropriate insight.
In order to provide you with appropriate information, I would need to know the nature of your organization and how your lab fits into it. But in general, I would say that your greatest challenge will be documentation. If you have an equipment log for your SEM and EDS (as you should), you have completed one aspect already. But in addition:
1.) You will probably need to generate some appropriate standard operating procedures (SOP's) for your equipment, including calibration and maintenance procedures. You should pay particular attention to how you schedule these, so that you will ensure compliance. Also, do not forget your standards; they are also subject to recalibration schedules.
2.) Sample traceability may be an issue for you as well. Be sure to have procedures describing how you handle and track them.
3.) If you have subcontractors who perform certain calibrations for you (e.g. picoammeters and stuff) you may need to have a document explaining how you assess them.
4.) Get some calibration stickers - you will need to tag any equipment that is subject to calibration. Equipment not requiring calibration may need reference only tags.
5.) You may need to conduct an equipment and software validation that includes installation, operation and processing (IQ, OQ, PQ) depending upon how the equipment is used. This can be quite extensive and time-consuming.
THE IMPORTANT THING TO REMEMBER WITH DOCUMENTATION IS THAT YOU DO WHAT YOU SAY YOU WILL DO. DO NOT GENERATE "ALL INCLUSIVE" DOCUMENTS THAT INCLUDE STEPS THAT YOU DO NOT NORMALLY DO - UNLESS YOU NEED TO CHANGE AND YOU PLAN TO STICK TO IT.
There are many other considerations that are too extensive to go into here, but if you still need information or would like to discuss this more, feel free to contact me direct.
Regards,
Bob Citron Chiron Vision Claremont, CA USA Bob_Citron-at-cc.chiron.com
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Our facility is undergoing ISO certification. My EM lab is small, consisting of one SEM with BSE and EDS. I would appreciate any suggestions/warnings, etc. from your experiences with ISO certification.
Majid: It is known that Spurr's resin has some incompatibilities for example I have seen problems using Kellenberger en bloc uranyl acetate staining with Spurr's resin. If this is the case it seems to affect some areas more than others in the block and is not due to mixing or infiltration. With proper usage Spurr's can infiltrate plant cells and moon rocks so it shouldn't be that hard to infiltrate cells in a monolayer. bob
Kenneth JT Livi wrote: =================================================== Now that the subject of holey-C grids has been raised, are there any vendors willing to guarantee contamination-free films? Most of the grids I've purchased from vendors will contain some silicon contamination, along with variable Na, Ca, K and Cl. Since I normally investigate silicates, Si contamination is very problematic. I assume the Si comes from diffusion pump oil. Have any of the vendors with "TEM quality control" checked for the cleanliness of their grids? =================================================== It is a correct statement that it is easy for grids to get contaminated as indicated. There is a reality check relating to cost, that is, we ourselves do not routinely check with EDS, each batch for the presence or absense of Si. But we do check periodically and will when specially requested, do a special batch check by EDS. This higher level of control represents slightly higher costs, that being the reason it is not done for every batch . If good housekeeping and cleanliness conditions are maintained, one can make "Si-free grids". In any case, we can produce what we ourselves determine (and believe) to be "Si-free" filmed grids as determined by our own EDS measurements on our in-house JEOL 100CX TEM. However, I am concerned about the "willing to guarantee" requirement. If you are in agreement that a "zero" amount of anything does not exist, then what detection limits are we talking about?
We have found at least a few of our customers probably have detection limits lower than what we are able to achieve when we do the control work. Clearly a small spot size XPS might find all kinds of things that would otherwise be "guaranteed to be free of" when done by EDS in a TEM.
There is another element to this equation: Some users "find" Si in our spectroscopically pure carbon mounts. We are convinced there is no silicon in our carbon mounts, at least if present at all, the level would be less than 1 ppm as determined by emission spectroscopy. Yet we are told by customers that Si is sometimes "detected". However, others have told us that the culprit is the Si in the Si (Li) detector being used. We know that at least some of the EDS systems (e..g like the older ones we have in house) seem to have some problem trying to model the "notch" for the background subtraction algorithms. So we have been operating on the theory that at least some of the so-called Si being detected in filmed grids is an artifact of the system being used, yet on the other hand, I am not saying it is always an artifact. After all, one CAN indeed end up with Si on filmed grids! The common practice of using silicone fluids in a vacuum evaporator is one already mentioned potential source.
The other comment would be that if one wants Si-free films, then to be on the safe side, at least we have for some time, used Santovac 5 in the diffusion pumps of the vacuum evaporators.
I hope that explains why we believe our holey carbon (filmed) grids are indeed "silicon-free". I would also expect that any other producer, taking the same precautions, should end up with grids that would be comparable.
I would welcome any further suggestions on this topic, either on-line or off -line.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ================================================== ------- FORWARD, End of original message -------
Although I work in medical research EM I am currently attending, in my spare time and for my own personal interest, a course in Horticulture. Part of that course involves the study of photosynthesis. Our instructor on the course does not have an EM photograph of a chloroplast which he can use as a "handout" for the class. I have no botanical EM material at all.
I was wondering if there is a kind soul on the List who would like to donate a scanned copy of a chlorplast. I can promise that it would be used for educational purposes only and not for any commercial purpose. Due acknowledgment would be printed on the image.
Ideally what I would like would be a JPEG, GIF or TIFF file which could be printed out on a decent laser printer and then the instructor could run off enough copies for the students on the course (about 50).
Failing that, could anybody with the relevant knowledge point me to a botany related website where I could download a public domain image of a chloroplast.
Quantitative Electron-Probe Microanalysis (Ellis Horwood Series in Physics and Its Applications) ~ Ships in 2-3 days Love G., et al / Paperback / Published 1995 Their Price: $77.00
Regards, Neal
Spaulding, Robert F wrote: } } I am looking for a source for the book, Quantitative Electron-Probe } Microanalysis by V. D. Scott and G. Love. My local bookstore is having } no luck. } } Thanks, } Robert F. Spaulding } Corning Incorporated } Characterization Science & Services } Microscopy & Microanalysis Dept. } SP-FR-01-8 } Corning, New York 14831
I am looking for a commercial lab in the New England area that I could send some samples out to be embedded, sectioned and TEM prints taken. If I could get the names of the labs and people to contact, I would appreciate it greatly. Please contact me off of the listserve so as not to infringe on those who are not interested. I will gladly supply more details about the samples to those who contact me.
David Bell Millipore Corporation phone: (781) 533-2108 fax: (781) 533-3143 e-mail: David_Bell-at-Millipore.com
You may visit my website at {http://www.ualberta.ca/~mingchen/pcell.htm} for the TEM image of a plant cell which has some chloroplasts in it. Also you should be able to print it out on your printer.
Best wishes,
Ming
On Thu, 9 Oct 1997, Stephen Griffiths wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hi } } Although I work in medical research EM I am currently attending, in my } spare time and for my own personal interest, a course in Horticulture. Part } of that course involves the study of photosynthesis. Our instructor on the } course does not have an EM photograph of a chloroplast which he can use as } a "handout" for the class. I have no botanical EM material at all. } } I was wondering if there is a kind soul on the List who would like to } donate a scanned copy of a chlorplast. I can promise that it would be used } for educational purposes only and not for any commercial purpose. Due } acknowledgment would be printed on the image. } } Ideally what I would like would be a JPEG, GIF or TIFF file which could be } printed out on a decent laser printer and then the instructor could run off } enough copies for the students on the course (about 50). } } Failing that, could anybody with the relevant knowledge point me to a } botany related website where I could download a public domain image of a } chloroplast. } } With thanks in advance } } Regards } Stephen Griffiths. } } {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} } Stephen Griffiths } Visual Science Department } Institute of Ophthalmology } Bath Street, London. EC1V 9EL } e-mail:- s.griffiths-at-ucl.ac.uk (work address) } or stephen.griffiths-at-dial.pipex.com (home address) } {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} }
*********************************************** * Ming H. Chen, PhD * * Medicine/Dentistry Electron Microscopy Unit * * University Of Alberta. * * Edmonton, Alberta, Canada * * * * Visit My Page At: * * http://www.ualberta.ca/~mingchen * ***********************************************
Kenneth JT Livi wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Now that the subject of holey-C grids has been raised, are there any } vendors willing to guarantee contamination-free films? Most of the grids } I've purchased from vendors will contain some silicon contamination, along } with variable Na, Ca, K and Cl. Since I normally investigate silicates, Si } contamination is very problematic. I assume the Si comes from diffusion } pump oil. Have any of the vendors with "TEM quality control" checked for } the cleanliness of their grids? } } Ciao for now, } Ken } } Kenneth JT Livi } Department of Earth and Planetary Sciences } 34th and Charles Streets } Johns Hopkins University } Baltimore, Maryland 21218 USA } Phone: (410) 516-8342 } Fax: (410) 516-7933 } e-mail: klivi-at-jhu.edu
Dear Kenneth Livi,
Kenneth JT Livi Wrote -
During the production of carbon film grids there are three (3) possible silicon contamination sources: 1) Residue in the evaporator 2) Evaporator Oil 3) Carbon Source
When we produce "silicon free" film we take the following precautions: 1) We thoroughly clean the evaporator prior to each evaporation cycle.
2) On all our evaporators silicon free oil is used.
3) We use pure carbon not graphite as a source. The impurity level does not exceed 2ppm
Since there is always a potential for silicon in the carbon the term "silicon free" is subjective but based on the above we feel we produce film with at most 1 ppm Si but it is impossible for LADD to guarantee the total absence of Si. :)
John Arnott PH: 802/878-6711 LADD RESEARCH FAX: 802/878-8074 13 Dorset Lane Williston, VT 05495
Dear all, I greatly should appreciate informations on experiences with existing/ or on dealers/companies of LASER PROJECTION SYSTEMS (connectable to PC/Video and TV Cams, Remote system). We know a company (ASK-System) in Europe which deals with LIGHT PROJECTION SYSTEMS for demonstration of slides, PC-data, etc. in teaching and else applications. Has anybody suggestions, informations (company/-ies, approx. price, combinations with periphery, necessary components) for us ? Such a Laser Projection System should work for projection screen dimensions of at the maximum approx. 2 x 3 m or little less, if available. Any suggestions and comments are welcome. Best wishes for the day sincerely yours Wolfgang MUSS Dept. Pathology LKA, EM-Lab, Muellner Hauptstrasse 48 A-5020 SALZBURG, Austria/Europe Phone: ++43++662+4482-4720 Ext Fax: ++43++662+4482-882 Ext. e-mail: W.Muss-at-lkasbg.gv.at.
Does any one have any image processing and analysis scripts/macros/programs they are willing to share regarding the processing and analysis of cross section neurons. to include axon thickness, sheath size as well as to classify and count cross section samples based on size.
I would be willing to share one program I have (written for Kontron KS400).
Now that the subject of holey-C grids has been raised, are there any vendors willing to guarantee contamination-free films? Most of the grids I've purchased from vendors will contain some silicon contamination, along with variable Na, Ca, K and Cl. Since I normally investigate silicates, Si contamination is very problematic. I assume the Si comes from diffusion pump oil. Have any of the vendors with "TEM quality control" checked for the cleanliness of their grids?
Janet Woodward asked about laboratory experiences with ISO issues. I am laboratory director of Structure Probe, Inc., which has been accredited since 1980 in the subdiscipline of "microscopy" by the American Association for Laboratory Accreditation. We are currently accredited to the standard of ISO Guide 25, which is the international document which specifically applies to laboratories. A direct comparison with ISO 9000-series documents shows that the requirements are roughly the same, plus Guide 25 requires a demonstration of competence, which is not required by ISO 9000. While our experience has been evolutionary, and Janet is about to jump into the deep end of the pool without having had swimming lessons, some of the things which we have learned over the years might help.
1. Read the Directions: Unfortunately, there are a number of very specific requirements, and it is important that you know what they are. It is one thing to be involved in a discussion about whether what you do does or does not meet a particular requirement--it is another to be "blindsided" by a requirement of which you were not aware.
2. Documentation: The biggest problem for us in the evolution of our accreditation experience is the need for explicit procedure documents which cover every aspect of our technical operations. Writing procedures for the assembly of widgets from boxes of parts can be a challenge--writing procedures for doing contract microscopy in an independent laboratory environment, where the nature of the work is totally driven by the requirements of clients, can be a nightmare. We have written documents that are very heavy on such issues as how to document each and every micrograph so that it can be unequivocally traced to a specific sample and management review of each and every incoming project. At the same time, the documents allow a great deal of professional judgement for the individual analyst, provided that the procedures actually used are documented in the report.
3. Calibration: This is the "culture shock" area. You will be expected to support your results with a calibration trail, and that calibration will be expected to be traceable, where possible, to a national standards body. I have found it helpful to use the word "calibration" but think about "verification". We have a program in which each column instrument is run through a set of standard evaluations every month, and the results are recorded as micrographs and spectra in log books. As a result, we can go back and show you the performance history (magnification, resolution, etc.) of the instrument and, if push comes to shove, we can answer the critical question--"How do you know that, on October 5, 1895, the magnification of this micrograph was in fact 3,000X?"
4. Traceable Reference Samples: The actual core of the calibration program is the repetition and documentation, but the problem that seems to be causing the most pain is the traceability of the reference samples used. This is a moving target. What is going on is that there is no such thing, as this is written on October 9, 1997, as an accredited calibration laboratory for standards for microanalysis. There are materials available from the National Institute for Standards and Technology (NIST) in the U.S. and from comparable bodies in other countries, but they do not cover many of the areas which a good calibration program will include. Various suppliers, including SPI Supplies (tm), offer reference materials suitable for calibration programs, but these are not necessarily fully traceable--if someone says that a reference material is traceable, ask what the "uncertainty budget" is (get used to that term, it covers the fact that every time you extend a measurement from the ultimate reference, you become less and less certain what the "actual" value is).
I am happy to discuss any of these issues in detail--off line is probably better. I am also happy to conduct dialog with your assessor--some of these problems are more easily resolved by some who can speak "quality".
Disclaimer: I am employed by Structure Probe, Inc., parent company of SPI Supplies. I am also a member of the Accreditation Council of the American Association for Laboratory Accreditation.
Andy
Andrew W. Blackwood, Ph.D. Structure Probe, Inc. P.O. Box 656 West Chester, PA 19381-0656 Ph: 1 610 436 5400 FAX: 1 610 436 5755 e-mail: ablackwood-at-2spi.com WWW: http://www.2spi.com
I am trying to find the mailing addresses, email addresses, phone or fax for the following electron microscopists. I want to reproduce figures from their books in our web course and the publisher has informed me that the copyright has reverted back to the author. Unfortunately, the publisher (Elsevier) no longer has addresses for these authors. If you can help me, email me at gsosinsky-at-ucsd.edu.
1) Alan W. Agar 2) Ronald H. Alderson 3 Dawn Chescoe 4) J.H. Martin Willison 5) Arthur J. Rowe
I am looking for advice from those experienced with using Bouin's fixative.
A users of our lab is about to leave on a cruise and needs to preserve plankton at sea. In the past he has used Bouin's fixative, but now, due to the picric acid component, the hassle factor has gone way up.
He is looking for either a pre-made Bouin's so he does not have to deal with the picric acid issue, or a substitute fixative that would serve his needs.
If you can help, send the information to me and I will pass it along to him.
Thanks.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
The Materials Research Science and Engineering Center (MRSEC) of the University of Maryland is looking for a post-doctoral fellow to perform research on ferroelectric and ferromagnetic oxide films using TEM. The MRSEC has very strong experience in the synthesis and characterization of these materials. We also have experitise in device fabrication. The position is available imediately with a starting salary of $32-35K depending on experience.
Experience on high resolution TEM is highly desirable. Interested candidates should submit their resume and a list of three references to Dr. Lourdes Salamanca-Riba either by regular mail or e-mail at either of the following addresses.
Materials and Nuclear Engineering Department University of Maryland College Park, MD 20742-2115
Thank you for your helpful responses to my question:
} What is the best commercial source of consistently } high-quality holey grids?
Ian Montgomery recommeds Agar Aids in England. He has been using the same grid for almost 30 years! That's my kind of quality production. British engineering is second to none. Ian, is that grid made of copper or kryptonite? ;-)
Larry Allard recommends grids produced by Structure Probe and Pella.
Brian Demczyk recommends SPI grids.
Markus F. Meyenhofer wrote "It's the fool behind the tool!!!". Thank you, Markus, for a touch of humor.
Representatives of Scott Scientific, Ted Pella, SPI Supplies, and Ladd Research wrote to praise their own products.
Does anybody have (good or bad) experience with any of the following or other fast freezing devices: Reichert KF80 Leica CPC Gentleman Jim (constructed by Alan Boyne)
Has anybody made a comparison?
Philip
-- Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 86 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www_scem.csb.ki.se/pages/philip.html
I'm sending this message on behalf of the Department of Biology at Eastern Connecticut State University. Our microbiologist had to undergo emergency surgery and is not expected to return to his teaching duties for another 5 to 8 weeks. We have been able to cover most of his classes, but right now his introductory microbiology class has no instructor. The lecture meets M, W, and F from 9 to 10:00 AM and the lab meets on Thursday from 2 to 5:00 PM. We are in desperate need of a temporary replacement. If anyone is interested or knows of someone, please let me know. Of course, the individual would have to live within reasonable driving distance of the University (Willimantic, Connecticut). If these times don't fit your schedule, but you are still interested, let me know. We might be able to rearrange the time.
If we cannot find a replacement, we are open to suggestions. Perhaps someone knows of a course on tape or even an ongoing internet course.
Please respond immediately via Email. I will forward your message to our department chair.
Thanks,
Martin A. Levin Department of Biology Eastern Connecticut State University Willimantic, CT 06226 Phone: (860)465-4324 FAX: (860)465-5213
We would like control our microscope Philips 420/STEM by connecting it to a PC, Mac or Silicon Graphics. Can anybody give advice to make it effectively? Thanks. Stanislav Hucek Lab. of electron microscopy Institute of Parasitology ASCR Branisovska 31 370 05 Ceske Budejovice Czech Republic
O.k., resin wizards, I have a user looking a truely clear, i.e. water clear, resin for emmbedding some small objects (1-2 cm x 2 mm neoperene O-rings) for gifts as paper weights. Therefore sectionablity or LM / EM is not a factor but durability and hardness are - they would like "nice and hard" resin.
I realize that this is not a strickly scientific quiry - and I apologize to anyone offended by any triviality - but why not ask the experts, eh? After who among us hasn't emmbedded a cockroach, etc. and kept it on their desk?
Commercial vendors should feel free to respond.
Thank you.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"640K ought to be enough for anybody." -- Bill Gates, 1981
This the SILVER Scottish Microscopy Symposium will take place at the above = =0Avenue and the Organising Committee have arranged a Scientific Programme = which =0Awe hope will appeal to as many microscopists as possible. Here is = the finalised =0Aprogramme.
Low temperature strategies for immunocytochemistry: choice or compromise - = =0AJeremy Skepper, Cambridge
Combining stereology and for immunogold labelling in quantitative =0Aimmuno= electron microscopy of membranes - John Lucocq, Dundee
Confocal Microscopy with high light budget - Tony Wilson, Oxford, England
Green Fluorescent Protein (GFP) as a tag for plant virus movement studies -= Alison =0ARoberts, Dundee
Environmental Scanning Electron Microscopy - Dirk van der Vall, Eindhoven,= =0AHolland
Staining resin sections - why infiltration is (or should be) incomplete - R= ichard =0AHorobin, Sheffield
Efficient and unbiased 3D measurements in microscopy - Stereology - Vyvyan = =0AHoward, Liverpool, England
Minimal preparation procedures for SEM of botanical specimens - Stephan Hel= fer, =0AEdinburgh
Registration will be =A320. For more information either contact me or you c= an visit =0Aour web site for the latest information and also details of las= t years meeting. Web site- http://www.abdn.ac.uk/~nhi691/smg97.htm=20 (This will be updated next week)
Kevin Mackenzie Tillydrone E.M. Unit University of Aberdeen Tillydrone Avenue Aberdeen AB9 2NT
This the SILVER Scottish Microscopy Symposium will take place at the above = =0Avenue and the Organising Committee have arranged a Scientific Programme = which =0Awe hope will appeal to as many microscopists as possible. Here is = the finalised =0Aprogramme.
Low temperature strategies for immunocytochemistry: choice or compromise - = =0AJeremy Skepper, Cambridge
Combining stereology and for immunogold labelling in quantitative =0Aimmuno= electron microscopy of membranes - John Lucocq, Dundee
Confocal Microscopy with high light budget - Tony Wilson, Oxford, England
Green Fluorescent Protein (GFP) as a tag for plant virus movement studies -= Alison =0ARoberts, Dundee
Environmental Scanning Electron Microscopy - Dirk van der Vall, Eindhoven,= =0AHolland
Staining resin sections - why infiltration is (or should be) incomplete - R= ichard =0AHorobin, Sheffield
Efficient and unbiased 3D measurements in microscopy - Stereology - Vyvyan = =0AHoward, Liverpool, England
Minimal preparation procedures for SEM of botanical specimens - Stephan Hel= fer, =0AEdinburgh
Registration will be =A320. For more information either contact me or you c= an visit =0Aour web site for the latest information and also details of las= t years meeting. Web site- http://www.abdn.ac.uk/~nhi691/smg97.htm=20 (This will be updated next week)
Kevin Mackenzie Tillydrone E.M. Unit University of Aberdeen Tillydrone Avenue Aberdeen AB9 2NT
The Department of Materials at Oxford University will shortly be setting up an Advanced Analytical HREM Facility, funded by a major grant from EPSRC. This will be based on a JEOL 3000F (300kv FEG/TEM) with a comprehensive range of analytical techniques, including EDX, GIF holography, etc. It is anticipated that the Facility will be set up early in 1998 to extend the present extensive range (12) of TEM and SEM instruments and support facilities. We are planning to appoint a Senior Research Officer who will be responsible for the day-to-day running of the Facility, and who will develop a successful Outside Users' programme. This will be a key position, funded for three years in the first instance. The successful candidate will be someone with expertise in the above techniques and some relevant post-doctoral experiance. Salary will be in the range 15159-22785 pounds sterling depending on age and experience.
For further details, please contact directly: Dr. John Hutchison, Department of Materials, University of Oxford, ParksRoad, Oxford. OX1 3PH. UK. phone: +44 1865 273705 e-mail: john.hutchison-at-materials.ox.ac.uk
I have been searching for an electronic overlay font (preferably a Microsoft compatible "True Type" variety rather than an Adobe "Post Script") to facilitate labeling of gray-scale images. Is anyone aware of a source for such work saver?
In the pre-digital days, we were getting good results with the paste-on overlay letters that are still available from microscopy supply houses (e.g., I was partial to the Microscopist's Collection from SPI). These were the little black letters that were printed over slightly larger white outlines. Because of this contrasting outline, they were easily readable irrespective of the brightness of the image below.
Currently, we are barely getting by in labeling of electronic images by adjusting the font color to either white or black, according to what the brightness of the underlying image dictates. More often than not, this is ineffective in EM images with highly modulated brightness. On occasion, I have resorted to manually overlaying a black text over a white one in a bold version of the same font; this is tedious and does not work in all but the briefest annotations because the kerning of the two font styles does not compensate for the differences in the widths of the letters.
The simplistic solution of setting the overlay text box background color to non-transparent is not satisfactory because it often ends up obscuring the feature of interest.
Does anyone possess a suitably outlined font, or has come up with a creative solution to this vexing problem?
valdemar-at-fast.net Valdemar Furdanowicz Homer Research Labs Bethlehem Steel Co. Bethlehem, PA
} We would like control our microscope Philips 420/STEM by connecting } it to a PC, Mac or Silicon Graphics. Can anybody give advice to make } it effectively? } Thanks.
I don't know specifically about the 420/STEM, but we're using the 4pi SE-II board in a Power Mac to control our Phillips STEM.
Imageing plug-ins work with NIH-Image or Photoshop. EDS plug-ins work with (NIST) DTSA or FLAME.
We're at work on a library to call 4pi driver routines from XlispStat ( a graphical statistics package ), Python (and interpreted object-oriented language), Java and NIH-Image. ( We've been using prototypes of these long enough to finally figure out how to redo them right! ;-)
See: http://www.4pi.com
---| Steven D. Majewski (804-982-0831) {sdm7g-at-Virginia.EDU} |--- ---| Department of Molecular Physiology and Biological Physics |--- ---| University of Virginia Health Sciences Center |--- ---| P.O. Box 10011 Charlottesville, VA 22906-0011 |--- All power corrupts and obsolete power corrupts obsoletely." - Ted Nelson
The "Analytical Methods" section of the following article includes operating conditions for electron microprobe analysis of Na-bearing zeolites. Analcime is not specifically addressed, but the described method should work for this mineral, too.
Ross, M., Flohr, M.J.K., and Ross, D., 1992, Crystalline solution series and order-disorder within the natrolite mineral group: American Mineralogist, v. 77, p. 685-703.
Back in the 1950s, the standard embedding medium for biological EM was a mixture of methyl and n-butyl methacrylates (~1:9), catalyzed with 2% Luperco CDB, and polymerized at 60oC (or by UV). That material (otherwise known as "plexiglass") was very clear and hard, and would satisfy your criteria. In fact, about 1958 I embedded a ~3.5" cockroach in methacrylate. I had encountered it in the hallway outside our lab (it looked like an approaching Volkswagen "beetle"). It was an escapee from another lab that was doing research on the nervous system of such critters.
Kent
A. Kent Christensen Department of Anatomy and Cell Biology Medical Sciences II Building University of Michigan Medical School Ann Arbor, MI 48109-0616 akc-at-umich.edu Tel (313) 763-1287 http://www.umich.edu/~akc/
------------------------------------
On Fri, 10 Oct 1997 edelmare-at-casmail.muohio.edu wrote:
} O.k., resin wizards, I have a user looking a truely clear, i.e. } water clear, resin for emmbedding some small objects (1-2 cm x 2 mm } neoperene O-rings) for gifts as paper weights. Therefore } sectionablity or LM / EM is not a factor but durability and hardness } are - they would like "nice and hard" resin. } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 352 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu
I am looking to buy or borrow a used power supply for a 100 W Hg arc lamp (specifically the Osram 100 W/2, although I believe most DC arc lamp supplies will work). Please respond to
umbach-at-msc.cornell.edu
or reach me by phone at 607-255-7426
Thanks!
-- Dr. Kit Umbach Dept. of Materials Science and Engineering 360 Bard Hall Cornell University Ithaca, NY 14853
we want to open the .img images of our XL20 and need some program for this purpose. Every hint appreciated.
Thanks in advance.
Andreas Loewe
______________________________________________________________ Andreas Loewe Tel: +49-228-734-180 University of Bonn Fax: +49-228-734-205 Institute for Inorganic Chemistry email: loewe-at-uni-bonn.de Inorganic Material Research Roemerstr. 164 53117 Bonn Germany http://www.elmi.uni-bonn.de/ ______________________________________________________________
} I am looking for advice from those experienced with using Bouin's fixative. } } A users of our lab is about to leave on a cruise and needs to preserve } plankton at sea. In the past he has used Bouin's fixative, but now, due to } the picric acid component, the hassle factor has gone way up. } } He is looking for either a pre-made Bouin's so he does not have to deal } with the picric acid issue, or a substitute fixative that would serve his } needs. } } If you can help, send the information to me and I will pass it along to him. } } Thanks. } } Jonathan Krupp
Substitute: I've used Trump's, Karnovsky's and plain old 2% glut or 10% formalin in 0.1 micron filtered seawater for plankton. Do you guys have a copy of the UNESCO book on Collection and Preservation of Zooplankton? (This includes ciliates, etc.) Try John Pearse, he might have one.
One solution is to import your image files into Microsoft Word (other word processors may have similar functions) and use the WordArt tool to overlay your picture with the desired text. This tool will allow you to outline letters using most true type fonts and allows you to control pitch, text color, outline color, and outline thickness. This method is fairly quick and painless. One hint is that when you first start adding text it may disappear behind the image. To bring the text to the front click on the image with the second mouse button, click on "order" and select "send behind text". This will place the image behind the text so that the text remains visible.
You can then either save the image in Word format or use a screen capture program to save the image in other formats. If using a screen capture program (such as LView) you will be limited to the resolution of your screen minus window borders so this solution works only for smaller images (aprox. 970x570 pixels on a 1024x768 monitor). There may be a way to export larger images from Word but as I am a novice to using Word I haven't found one yet.
Dan Moore
At 12:17 PM 10/10/97 -0400, you wrote: } Dear Colleagues: } } I have been searching for an electronic overlay font (preferably a } Microsoft compatible "True Type" variety rather than an Adobe "Post } Script") to facilitate labeling of gray-scale images. Is anyone aware of a } source for such work saver? } [snip] } } Does anyone possess a suitably outlined font, or has come up with a } creative solution to this vexing problem? } } valdemar-at-fast.net } Valdemar Furdanowicz } Homer Research Labs } Bethlehem Steel Co. } Bethlehem, PA } }
I should have added that using Word you can rotate your labels and add arrows, circles, boxes, etc. to your images. You can even add cartoon balloons and let your subjects speak to you.
Vlademar, There is a software package called "Designer" it is sold by Micrografx. It is very powerful and will do what you need. The cost is about $600. It is worth while to check it out.
Greg Rudomen Greg-at-umic.sunysb.edu S.U.N.Y. Stony Brook University Microscopy Imaging Center 516-444-3126
valdemar wrote:
} I have been searching for an electronic overlay font (preferably a } Microsoft compatible "True Type" variety rather than an Adobe "Post } Script") to facilitate labeling of gray-scale images. Is anyone aware of a } source for such work saver? } } In the pre-digital days, we were getting good results with the paste-on } overlay letters that are still available from microscopy supply houses } (e.g., I was partial to the Microscopist's Collection from SPI). These } were the little black letters that were printed over slightly larger white } outlines. Because of this contrasting outline, they were easily readable } irrespective of the brightness of the image below. } } Currently, we are barely getting by in labeling of electronic images by } adjusting the font color to either white or black, according to what the } brightness of the underlying image dictates. More often than not, this is } ineffective in EM images with highly modulated brightness. On occasion, I } have resorted to manually overlaying a black text over a white one in a } bold version of the same font; this is tedious and does not work in all } but the briefest annotations because the kerning of the two font styles } does not compensate for the differences in the widths of the letters. } } The simplistic solution of setting the overlay text box background color to } non-transparent is not satisfactory because it often ends up obscuring the } feature of interest. } } Does anyone possess a suitably outlined font, or has come up with a } creative solution to this vexing problem?
This is the recipe that I use in Photoshop 4.0 to put Black on White scale markers, text, and symbols onto micrographs. The results look just like the rub-on transfers that I used to use.
1. create a layer (Photoshop 4.0 does this automatically with text tool) You must use Photoshop image mode for the layer option.
2. in that layer in the font and font size that you want, type the text. add a black line at an appropriate length and width and any other text, symbols, arrows, etc. that you want to put on the micrograph. By using the layer, you preserve the original micrograph in the background layer. you can use the info window to draw lines to particular lengths. If other layers are created when new text is added, merge those layers. Don't merge them with the background layer!
3. Select all (ctrl-A in the PC) The marquee will be around the whole layer.
4. You have to move the selected region up then down with an arrow key. (This is done in the PC with the Ctrl-shift-arrow key in the PC) what this does is to select all of the objects in the layer individually. A Marguee should be around each object.
5. Select the foreground color as white. (You are going to write a white border around each Marquee.)
6. Go to Edit-Stroke and select the width of the white line you want (Width) and select the Outside option. for 300 dpi images at about 4" x 5", I suggest a font size of about 14 (Arial) with a Width of about 3-4 pixels for the stroke width. This will write a white border 4 pixels wide around all of the selected black features.
7. Deselect (ctrl-D)
8. If you want to save this as image in another format such as TIF or BMP, then you have to Merge the layers and save the image in that mode.
Note: you should have anti-aliasing selected for all this.
This technique works very well for me. You can make them look like real Rub-ons with another technique and offsetting the white a little, but that is another story.
Scott D. Walck PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of S.D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
Philip, I used a Gentleman Jim Plunge Freezer a million years ago and as I recall our tissue had freeze damage.
For a review try: Ryan, KP (1992) Cryofixation of tissues for electron microscopy: A review of plunge cooling methods. Scanning Electron Microsc., 6:715-743. I got that reference from Scott Russell's article (1997) Spray-Freezing Freeze Substitution of Cell Suspensions for Improved Preservation of Ultrastructure. Microscopy Research and Technique 38:315-328. He lists several references for freezing device reviews.
best regards, beth
************************************** Beth Richardson EM Lab Coordinator Botany Department University of Georgia Athens, GA 30602
Dear all,=20 (according to a suggestion of Scott WHITTAKER I place the posting to the = server once more) if there anybody still is out there interested in getting written = informations on "How do do it", product data sheets (in English or = German, if you like) of the silicone rubber used, the adress of the = Company (in the USA and Germany) and a sample mold out of my fabrication = (all free of charge)=20 PLEASE SEND an E-MAIL NOW (to get it from my desk) to:
W.Muss-at-lkasbg.gv.at (note: the "l" right to "-at-" is a small "L").
Thank you for your interest.=20 Best regards, have a nice and a calm weekend, W.MUSS
} I have been searching for an electronic overlay font (preferably a Microsoft } compatible "True Type" variety rather than an Adobe "Post Script") to } facilitate labeling of gray-scale images. Is anyone aware of a source for } such work saver? } } In the pre-digital days, we were getting good results with the paste-on } overlay letters that are still available from microscopy supply houses (e.g., } I was partial to the Microscopist's Collection from SPI). These were the } little black letters that were printed over slightly larger white outlines. } Because of this contrasting outline, they were easily readable irrespective } of the brightness of the image below. } (snip) } Does anyone possess a suitably outlined font, or has come up with a creative } solution to this vexing problem?
We use Adobe Photoshop. In version 4.0, be sure to select the outlined text tool so that the letters are surrounded by marquees when you are done typing. Type your text on the image. Before you de-select the text, go to the pallette region of the toolbar and exchange foreground (usually black) and background (usually white) colors. Then go to the "Edit" menu and select "Stroke" chose a 2-3 pixel stroke width "outside" the letter. This makes an outlined text in any font on your system.
-- Best Regards, John Minter
Eastman Kodak Company Phone: (716) 722-3407 Analytical Technology Division FAX: (716) 477-3029 Room 2142G Bldg 49 Kodak Park Site email: minter-at-kodak.com Rochester, NY 14562-3712 calendar: via PROFS
Seemed like PhotoStyler used to have an option for "drop shadowing". I would assume that PhotoShop also would have that eature. you could chose the amount and direction of shift of the shadow. I think the colors were also cusomizable.
However, I don't have acopy of either very accessible just now. For as much as it comes in handy, I can't imagine such a feature being dropped. } } Dear Colleagues: } } I have been searching for an electronic overlay font (preferably a } Microsoft compatible "True Type" variety rather than an Adobe "Post } Script") to facilitate labeling of gray-scale images. Is anyone aware of a } source for such work saver? } } In the pre-digital days, we were getting good results with the paste-on } overlay letters that are still available from microscopy supply houses } (e.g., I was partial to the Microscopist's Collection from SPI). These } were the little black letters that were printed over slightly larger white } outlines. Because of this contrasting outline, they were easily readable } irrespective of the brightness of the image below. } } Currently, we are barely getting by in labeling of electronic images by } adjusting the font color to either white or black, according to what the } brightness of the underlying image dictates. More often than not, this is } ineffective in EM images with highly modulated brightness. On occasion, I } have resorted to manually overlaying a black text over a white one in a } bold version of the same font; this is tedious and does not work in all } but the briefest annotations because the kerning of the two font styles } does not compensate for the differences in the widths of the letters. } } The simplistic solution of setting the overlay text box background color to } non-transparent is not satisfactory because it often ends up obscuring the } feature of interest. } } Does anyone possess a suitably outlined font, or has come up with a } creative solution to this vexing problem? } } valdemar-at-fast.net } Valdemar Furdanowicz ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-8216
I am posting this for the principal investigator listed below. Please send all request and questions to the person at the address listed. Thank you
EXPERIENCED ELECTRON MICROSCOPIST
To participate in highly interactive and motivated lab of molecular biologists; analysis of gene knockout mice with emphasis on the cytoskeleton. Ph.D. is desirable; expertise in immuno EM is exxential. Long-term position. Submit r=E9sum=E9. and list of three references to:
Dr. Elaine Fuchs Howard Hughes Medical Institute The University of Chicago 5841 Sout Maryland Avenue MC1028 Room N314 Chicago, IL 60637 =46ax: 773-702-0141
Mei Lie Wong Department of Biochemistry HHMI-UCSF Ph. 415-476-4441 Fax 415-476-1902 email wong-at-msg.ucsf.edu
} One solution is to import your image files into } Microsoft Word (other word processors may have } similar functions) and use the WordArt tool
Why not import the image file into PhotoShop or Corel where there is a huge variety of text/drawing tools and little constraint on the size/resolution of the image file? You can do composites, layouts, cutouts, masks, etc. quite handily.
If you do, you will pellet your gold. As a matter of fact, this is one way to clean up your old reagents of protein that has come off.
On Thu, 2 Oct 1997, Pat Hales wrote:
} Date: Thu, 02 Oct 1997 09:34:31 -0700 } From: Pat Hales {hales-at-medcor.mcgill.ca} } To: microscopy-at-sparc5.microscopy.com } Subject: Gold Conjugated Antibodies } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } For years we have "airfuged" our fluorescent antibodies at 95 000 rpm before } use to eliminate any aggregates which may have formed during storage. Does } anyone know if this can (or should) be done with gold conjugated antibodies? } } TIA, } } Pat Hales } McGill University } Dept. of Anatomy & Cell Biology } hales-at-hippo.medcor.mcgill.ca }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Dear Richard, The nicest clear resin I have seen is one the geologists use called: transoptic. It is a hot-press resin that comes up hard and clear and in good contact with the material. I don't know any details of where to get it. You wrote: } O.k., resin wizards, I have a user looking a truely clear, i.e. } water clear, resin for emmbedding some small objects (1-2 cm x 2 mm } neoperene O-rings) for gifts as paper weights. Therefore } sectionablity or LM / EM is not a factor but durability and hardness } are - they would like "nice and hard" resin. } } I realize that this is not a strickly scientific quiry - and I } apologize to anyone offended by any triviality - but why not ask the } experts, eh? After who among us hasn't emmbedded a cockroach, etc. } and kept it on their desk? } } Commercial vendors should feel free to respond. } } Thank you. } } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 352 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu
Regards, Mary Mary Mager Electron Microscopist Metals and Materials Eng., UBC 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel:604-822-5648, fax:604-822-3619 e-mail: mager-at-unixg.ubc.ca
I am a graduate student in the MBA program at Rockhurst College in Kansas City, Missouri.
For a class I am taking this semester, I am required to conduct some marketing research. This research will determine the public's attitudes toward a variety of consumer products.
Your email address was randomly chosen by me to be invited to participate in this program. If you would like to cooperate, you will be asked to answer via email a series of questionnaires. The surveys can always be answered by using the "RESPOND" feature of your email software and filling in the blanks on forms that will be sent to you.
Each survey should take only a couple of minutes to complete. There will be a small non-monetary gift awarded by email to those who complete all the surveys on time.
To participate in this project you must: - Be at least 18 years old. - Be a citizen of the United States. - Have an email address that ends with .gov, .com, .org, .net, or .edu.
If you would like to help me with this project, please send an EMPTY email to jdeshon-at-sky.net with only the words "YES SURVEY" (without the quotes) in the SUBJECT line. I will email you further instructions.
If you don't want to participate, kindly ignore this message. If I hear nothing from you, I will place your email address on my suppression file and you will never be bothered by me again.
Whether you participate or not, I hope you have a wonderful day.
I'm wondering whether anyone has remote TEM capabilities working via Internet connections, similar to remote SEM facilities that I've seen and heard about. I was told that remote TEM was not currently available. I would like to know whether remote TEM is useful and/or practical.
Casey Lu Assistant Professor of Biology Humboldt State University
I'm planning on submitting a grant proposal to NSF this fall to obtain matching funding for a TEM at Humboldt State University, located in far northern California. One of the questions that I must answer is "Why teach undergraduates TEM?" TEM is obviously a major tool used by cell biologists, and biologists in general must regularly interpret TEM-based information, but do they really benefit from learning the techniques? I would argue that they do greatly benefit and point out that students of TEM learn how to design and carry out rather involved biological investigations. They must be careful, persistent, and use reasoning to determine whether real changes have occurred in tissues/cells under investigation. I'm wondering if there are other benefits for students who learn TEM, such as possessing a valuable skill for jobs in the drug industry etc.
I would greatly appreciate any thoughts on the value of learning TEM as an undergraduate biology major.
Casey Lu Assistant Professor of Biology Humboldt State University
Casey: check out the URL: http://tpm.amc.anl.gov/MMC.
Also, details about Remote Microscopy at the High Temperature Materials Lab at ORNL are given at http://www.ms.ornl.gov/htmlhome/mauc/MAGRem.html.
Larry PS who told you remote TEM was not currently available?
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
We start to work in the field of image analysis in application to the light microscopy of steels, aluminium, titanium and copper alloys. Our problems are standard: grain size measurement, distribution of grain size, area of phases %, and so on. We try to assemble the simple system on the base of PC (486DX100). Now we looking for the software for this problems, but financial situation in Ukraine is rather bad. We have the preliminary information about the existing of free ware and shareware programs for quantitative metallography and image analysis for IBM PC compatible computers. The not expensive commercial programs in this field are interested for us too. We need for Your advise: where we can obtain or buy this programs ? The most convenient way for us is to transfer the files via E-mail. Ordinary and express mail services (for example UPS or DHL) are accessible for us too. We are very grateful for Your help.
With best regards Vladimir Pashinsky
Ukraine (fSU), Donetsk State Technical University, Department of Material Science
We start to work in the field of image analysis in application to the light microscopy of steels, aluminium, titanium and copper alloys. Our problems are standard: grain size measurement, distribution of grain size, area of phases %, and so on. We try to assemble the simple system on the base of PC (486DX100). Now we looking for the software for this problems, but financial situation in Ukraine is rather bad. We have the preliminary information about the existing of free ware and shareware programs for quantitative metallography and image analysis for IBM PC compatible computers. The not expensive commercial programs in this field are interested for us too. We need for Your advise: where we can obtain or buy this programs ? The most convenient way for us is to transfer the files via E-mail. Ordinary and express mail services (for example UPS or DHL) are accessible for us too. We are very grateful for Your help.
With best regards Vladimir Pashinsky
Ukraine (fSU), Donetsk State Technical University, Department of Material Science
Software such as Image Pro and Materials Pro are available through us, Evex analytical or through your Image Pro Reseller in the Ukraine.
For more information contact Medica Cybernetics at www.mediacy.com
Cheers Peter Tarquinio Evex Analytical
----------
Dear colleagues,
We start to work in the field of image analysis in application to the light microscopy of steels, aluminium, titanium and copper alloys. Our problems are standard: grain size measurement, distribution of grain size, area of phases %, and so on. We try to assemble the simple system on the base of PC (486DX100). Now we looking for the software for this problems, but financial situation in Ukraine is rather bad. We have the preliminary information about the existing of free ware and shareware programs for quantitative metallography and image analysis for IBM PC compatible computers. The not expensive commercial programs in this field are interested for us too. We need for Your advise: where we can obtain or buy this programs ? The most convenient way for us is to transfer the files via E-mail. Ordinary and express mail services (for example UPS or DHL) are accessible for us too. We are very grateful for Your help.
With best regards Vladimir Pashinsky
Ukraine (fSU), Donetsk State Technical University, Department of Material Science
Casey Lu asked about remote control of TEM/SEM over the internet
In addition to the reference that Larry Allard gave to the DoE2000 Materials Microcharacterization Collaboratory (http://tpm.amc.anl.gov/mmc) which has links and connections to the TPM programs at ANL, LBNL, NIST, ORNL, & the University of Illinois. You should also get a copy of the proceedings of Microscopy & Microanalysis '96 meeting in Minneapolis. The proceedings were published by the society through the Journal of the Microscopy Society of America (http://www.msa.microscopy.com).
At that meeting there was an entire day long session on TelePresence Microscopy in Education and Research. This symposium covered nearly all aspects of TelePresence from TEM to SEM. You can obtain copies of most all of the abstracts via the MSA site (http://www.msa.microscopy.com) in Adobe Acrobat PDF format, to find these you must go to "Past and Future MSA meetings" on the MSA site and then use the "View an On-line Copy of the 1996 Meeting Abstracts" link. Specify the search phrase "TelePresence Microscopy". There are 13 papers listed there, 6 of which deal with TEM.
A follow-up symposium will also be held at the Microscopy & Microanalysis 98 meeting in Atlanta. The meeting is scheduled for July 12-16th so mark your Calendar if your interested.
Thanks to everyone who responded to my query about marking fluorescent TEM screens. Most related that it can be done, carefully, with a soft lead pencil. I tried it, it worked. Contact me off-line if you'd like more info.
Owen
Owen P. Mills Michigan Technological University Metallurgical & Materials Engineering Rm 512 MME Building Houghton, MI 49931 906-487-2002 906-487-2934 FAX opmills-at-mtu.edu
Has anyone performed image processing and analysis on nerve? Following is the question posed to me, I am clueless on what I would be measuring/doing regarding nerve analysis.
I do not have limited IA and EM experience with nerves would anyone recommend the best methods for fixation, and the types/concentration of fixatives used.
Input from Experts on neurology would be appreciated.
I suspect that there are probably reports of quantitative approaches to assessing peripheral nerves?? Does anyone have any suggestions?
Project:
I will be working with a diabetic drug, I believe designed to be an insulin "secretagog" (don't quote me on that) which in a previous mouse study caused a peripheral neuropathy morphologically similar to the classic spontaneous ageing neuropathy of mice. The hypothesis is that drug related changes in blood glucose contributed to the pathogenesis and thus it is a pharmacologic effect.
What questions should I be asking Gregory.Argentieri-at-pharma.novartis.com
---------- Von: Deutschlaender, Norbert, Path. An: Gregory.Argentieri-at-sandoz.com Betreff: AW: Image Processing and Analysis on Nerve Datum: Dienstag, 7. Oktober 1997 12:04
Dear Gregory, be careful not to mix up "diabetic neuropathy" (with hyperglycemia) and hypoglycemic damage. Isn't the drug you mention a compound producing hypoglycemia (followed by mainly cerebral alteration, i.e. in cortex and hippocampus; rarely in anterior horn neurons of spinal cord followed by axonal degeneration of peripheral nerves)? This is the first question you have to ask. Before having a clean qualitative neuropathologic diagnosis you better do not go into any morphometry of nerves (fiber density, fiber diameter, "roundness" of profiles, myelin sheet thickness etc.). At least in rats and humans the target for hypoglycemic damage is the central nervous system. Immersion of nerves with fixative (for instance Karnovsky's) turns mostly out to be adequate to perfusion, if you inhibit postmortal contraction (in situ fixation of nerve segment fixed to a wooden stick by 2 filament-knots before dissection, or at least fixation of segment attached to piece of firm paper).
Think further of doing fiber teasing, and using more distal nerve segments (tibial, sural nerve) instead of sciatic, because you may have a distally pronounced axonopathy in this model.
Norbert.
Dr. med. vet. Norbert Deutschlaender; Hoechst Marion Roussel Frankfurt/Germany. ---------- Von: Gregory.Argentieri-at-sandoz.com An: Gregory.Argentieri-at-sandoz.com; microscopy-at-Sparc5.Microscopy.Com Betreff: Image Processing and Analysis on Nerve Datum: Montag, 6. Oktober 1997 18:11
Greg, for your information, Rob Vogt at Erim (Environmental Research Institute of Michigan) has done extensive work in nerve resconstruction using Visilog. I believe he works on AXXON fibers or something similar.
If you want his phone or email, let me know. I can also fax you a paper that he has written on the subject if you are intereted.
Luc Noesis Vision Inc.
At 01:07 PM 10/12/97 -0500, Gregory.Argentieri-at-sandoz.com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
---------------------------------------------------------------------------- -------- Luc Nocente Tel: 514 345 1400 Noesis Vision Inc. Fax: 514 345 1575 e-mail: ln-at-noesisvision.com http://www.noesisvision.com 6800 Cote de Liesse, Suite 200 St-Laurent, PQ H4T 2A7,Canada ---------------------------------------------------------------------------- --------
We paste up our digital images onto pages using CorelDraw! CorelDraw! enables you to make an outline around your letters in a contrasting colour. So we can have black letters with a white minimal outline or vice-versa. It works with any typeface. And of course you can make the text any colour or shade of gray.
Mel Dickson Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
To convert XL.img files to tiff one can use either:
maketiff.exe (some Philips made program) DeBabelizer by Equilibrium opens .img files as well Graphicconverter by Lemkesoft (not sure)
Thanks again.
Andreas Loewe
______________________________________________________________ Andreas Loewe Tel: +49-228-734-180 University of Bonn Fax: +49-228-734-205 Institute for Inorganic Chemistry email: loewe-at-uni-bonn.de Inorganic Material Research Roemerstr. 164 53117 Bonn Germany http://www.elmi.uni-bonn.de/ ______________________________________________________________
} Dear colleagues, } } We start to work in the field of image analysis in application } to the light microscopy of steels, aluminium, titanium and } copper alloys. Our problems are standard: grain size } measurement, distribution of grain size, area of phases %, } and so on. We try to assemble the simple system on the } base of PC (486DX100). Now we looking for the software for } this problems, but financial situation in Ukraine is rather bad. } We have the preliminary information about the existing of free } ware and shareware programs for quantitative metallography } and image analysis for IBM PC compatible computers. The not } expensive commercial programs in this field are interested for } us too.
Dear Vlad,
there is quite a selection of shareware programs for the PC though I'm not sure how well they will perform on a 486. You might have to invest in a Pentium.
Try the following download sites:
Image Tool: http://ddsdx.uthscsa.edu/dig/itdesc.html ftp://maxrad6.uthscsa.edu ImageTool Mirror Sites Belgium - ftp://pa.cc.kuleuven.ac.be/pub/ImageTool Japan - ftp://gold.fish.kagoshima-u.ac.jp/pub/Win/science/ImageTool UK - ftp://micros.hensa.ac.uk/mirrors/uthscsa USA - ftp://wuarchive.wustl.edu/packages/graphics/image-tool
Osiris: http://www.expasy.ch/www/UIN/html1/projects/osiris/osiris.html OSIRIS software can be obtained free of charge from: Digital Imaging Unit University Hospital of Geneva 24 Micheli du Crest 1211 Geneva 14 - Switzerland Fax: (+41 22) 372 61 98 email : osiris-at-dim.hcuge.ch A special developper license is available for the full source code. Please contact us for more information.
Mage: ftp://suna.biochem.duke.edu/pub/PCprograms/
ScionImage: http://www.scioncorp.com ftp://scioncorp.com/ (the Macintosh program NIH image ported to the PC, but it seems to require a Pentium. Maybe You can get an older version for the 486.)
If You need any help, don't hesitate to ask.
Philip
-- Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 86 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www_scem.csb.ki.se/pages/philip.html
POSTDOC POSITION IN COMPUTATIONAL SOLID STATE SCIENCES
At Department of Physics, Norwegian University of Science and Technology (NTNU), Trondheim, Norway there is a vacant postdoc position for a two years period. The postdoc will work in the Electron microscopy group, NTNU and the starting date is December 1st, other dates can be discussed. The position is financed by the Norwegian Research Council under a joint program between Professor Ragnvald Høier NTNU; Professor Johan Taftø, University of Oslo; Dr. Bjørn Anderson, SINTEF Materials Technology, Oslo and Dr. Jon Samset, IFE, Oslo.
Applicants should have a background in solid state theory and preferentially experience with computer modelling. The postdoc will be involved in material modelling problems closely connected to ongoing experimental activity ranging from early stages of nucleation and growth of new phases in aluminium alloys to bonding and ductility in intermetallics and EELS interpretation. Further, in addition to general material characterisation the group at NTNU is in particular strongly involved in development and application of quantitative convergent beam electron diffraction.
The computing facilities are good (CRAYT3E and UNIX workstations).
Applications should be sent to Professor Ragnvald Høier, Department of Physics, Norwegian University of Science and Technology, N-7034 Trondheim, Norway as soon as possible and before Nov. 10th. Phone +47-73-593588 Fax +47-73- 597710 e-mail ragnvald.hoier-at-phys.ntnu.no
More information can be obtained from: Dr. Knut Marthinsen, SINTEF Materials Technology, N-7034 Trondheim, Norway e-mail: knut.marthinsen-at-matek.sintef.no or
Dr. Randi Holmestad, Department of Physics, Norwegian University of Science and Technology, N-7034 Trondheim, Norway e-mail: randih-at-phys.ntnu.no
I am forwarding a question to the list from a colleague who is not a member of the list. Please respond to his address as requested. Thank you.
Hello, I am seeking information on the use of Cationized Ferratin in experiments to study extracellular structures on cellulolytic bacteria (or any others for that matter). I believe that the nature of the stain is such that it can cause proteins to aggregate on the surface of bacteria and cause inaccurate readings. If you have any information either in support or disproving this idea please let me know. I would love to discuss my research, as we are trying to get a publication ready. I am not a member of this list so please send your responces directly to me at the address below.
Here is an updated and slightly amended version of the EM technician job which I mailed to the list recently.
Informal enquiries to me
Chris
UNIVERSITY OF MANCHESTER SCHOOL OF BIOLOGICAL SCIENCES
RESEARCH TECHNICIAN, GRADE C
(ELECTRON MICROSCOPY)
Applications are invited for the position of electron microscope technician within the School of Biological Sciences Electron Microscope, Graphics and Photography Unit. The post will involve working closely within a team of technicians in the Unit; providing assistance with electron microscopy operation, sample preparation, image processing, image archiving, data interpretation, networking and computer software management and maintenance. Applicants should have working experience in electron microscopy and preferably a working knowledge of computers. The salary for this post is the grade C scale stlg10,735 - stlg12,037.
Applicants should be qualified to a minimum ONC/2A - level standard and preferably hold an HNC/BSc in a biological sciences subject, physics, computation or mathematics.
Application forms and further particulars are available from Mr A. Nicholas, School of Biological Sciences, University of Manchester, 2.205 Stopford Building, Oxford Road, Manchester, M13 9PT, UK. email anichola-at-fs1.scg.man.ac.uk
The closing date for applications in October 31, 1997.
The University of Manchester is an equal opportunities employer
Chris Gilpin Biological Sciences Electron Microscope Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 161 275 5170 fax +44 161 275 5171 http://www.biomed.man.ac.uk/biology/emunit/emhome.html
I am physicist and so my SEM-EDX application field is inorganic material. I need to know whether a very small dried sample is blood. I have not any device to biological sample preparation. I have obtained the following EDX standardless semicuantitative analysis:
Unknow sample My own blood sample
C = 50.9 C = 47.4 O = 39.3 O = 44.6 Na= 0.5 Na= 1.2 Si= 0.1 Si { 0.1 (presence) P = 0.4 P = 0.2 S = 1.8 S = 1.4 Cl= 2.8 Cl= 2.7 K = 3.2 K = 1.6 Fe= 0.6 Fe= 0.4 Cu { 0.1 (presence) Cu { 0.1 (presence)
Are sufficient these data to conclude that the unknow sample is blood or furthermore human blood?. If not, what may I to do?.
Colleagues: Does anyone know of a good basic reference (textbook, tutorial, etc.) for fluorescence in situ hybridization (FISH)? Looks like I'm going into the FISH business, and I need to learn the basic techniques. Any help most humbly appreciated.
TIA-- Bev Maleeff SmithKline Beecham Pharmaceuticals Toxicology-US, UE 0462 709 Swedeland Road King of Prussia, PA 19406 610/270-7987 610/270-7202 fax Beverly_E_Maleeff-at-sbphrd.com
} I am physicist and so my SEM-EDX application field is inorganic } material. I need to know whether a very small dried sample is blood. I } have not any device to biological sample preparation. I have obtained } the following EDX standardless semicuantitative analysis: } } Unknow sample My own blood sample } } C = 50.9 C = 47.4 } O = 39.3 O = 44.6 } Na= 0.5 Na= 1.2 } Si= 0.1 Si { 0.1 (presence) } P = 0.4 P = 0.2 } S = 1.8 S = 1.4 } Cl= 2.8 Cl= 2.7 } K = 3.2 K = 1.6 } Fe= 0.6 Fe= 0.4 } Cu { 0.1 (presence) Cu { 0.1 (presence) } } Are sufficient these data to conclude that the unknow sample is blood or } furthermore human blood?. If not, what may I to do?. } } - Thank you in advance -
What is the sample? In what form did you get it? Are there square or cubical structures that are probably salt crystals (from a buffer solution)?
Have you examined the sample at low kV? {5kV, 1 or less better. You should still be able to see indications of (likely badly distorted) biconcave red cells (RBCs), and more likely platelets. Depending on how long the sample set before it dried, the platelets will be like tiny discs (smaller than the RBCs--say half the size or less, depending on how much the RBCs shrank) if it dried quickly. If the sample dried slowly, so the platelets were in fluid for a half hour or so, they will spread out and thin, with an irregular margin. They'll be larger in size, but very thin. 5kV is likely to be too high a voltage, likely you'll go right through them and not see the platelets. 1kV or less is needed. Examine your own blood sample first, starting with a fresh one, to find these structures, then go to the unknown. I'm not giving sizes, as there are too many variables for that to be meaningful, other than "smaller than RBCs", sizes of which in the SEM depends on how prepared and how dried (everything optimum, they're round about 8 microns in the SEM).
Any chance of getting the sample onto a formvar coated TEM grid, then putting that grid over a hole in the SEM stub? You'll have a better chance of seeing things if you're not imaging the substrate through the sample.
I am seeking information(micrographs, etc.) on the low-cycle-fatigue and high-cycle-fatigue fractures of ferritic malleable iron. The usual handbooks are of very little help. Thank you for any help.
My favorite program for creating and manipulating text is also CorelDraw. However, I just bought a package of plugins for use with Photoshop or Corel PhotoPaint. The package from Xaos (http://www.xaostools.com/products/index.html) cost $129 and included: TypeCaster, Paint Alchemy and Terrazo. The TypeCaster plugin will enable you to create highly visible text on your micrographs--it may be difficult to restrain your creative impulses. The other plugins may be useful for creating backgrounds for slides, web page images and as simple relief from the humdrum of scientific data.
I have no connection with Xaos financial, social or otherwise--wish I did! Larry D. Ackerman (415) 476-8751 Howard Hughes Medical Institute FAX (415) 476-5774 UCSF, Box 0724, Rm U426 533 Parnassus Ave. mishot-at-itsa.ucsf.edu San Francisco, CA 94143
Electron Probe filaments are just the same as are being used in the typic= al SEM. The difference is the position of the filament. If you place the filament a long way from the cap you need less heat because the bias fiel= d is more effective, the result is less evaporation, less emission current=
and a longer filament life. You do not need many electrons to generate enough x-rays for analysis compared with normal SEM imaging!
Saturation is saturation, it should be on the plateau of the graph. =
However the plateau may be moved higher or lower in the heat range depending upon the position of the filament and the amount of bias being used. I am afraid all those who have replied seem to be writing off electron gun importance, this is totally wrong, probe current is basicall= y number of electrons, get more from the gun and the current goes up. The gun sets the quality of an SEM image, set the gun up incorrectly and the rest of the system cannot compensate, you just run out of electrons!
Driving the filament hard means pushing it forward and increasing the bia= s field to constrain the beam but aiming in a Japanese instrument for 100 t= o 120uA emission current. This filament pushed forward increases the numbe= r of electrons being emitted from the cap, but this means more heat is required to reach saturation as more heat is lost to the cathode cap and because the bias field effect is weakened. More heat shortens the filame= nt life through evaporation. Increasing the bias constrains the electrons funnelling them together to try to achieve a small source. High performance requires a small high electron dense source which is improved=
further by using the correct anode-cathode distance of 1mm for every 2kV.=
Put very simply put a 50um source gives a 50A microscope, the condenser system giving about a 10,000X reduction. Reduce the source size but keep=
up the number of electrons it contains and you improve the instruments performance, 40um source gives a 40A microscope, it is possible to get mo= re than you paid for; the cost is filament life!
I can't vote positively for your results, although I am working in physical science as well.
In the range of my knowledge, a large error could be induced in the standardless EDS analysis for the light elements such as C and O, except you got a super-advanced detector and software to do the job. The results could be used to prove that the major contents of your sample are C and O, or possibly with H and N which you have not counted into the results or which are undetectable by the EDS.
The other reason I would not support your suggestion of the "human blood" is that the measurements of the minor contents such as Si, P, Na and Cu were less than 0.5%. They should be considered as the experimental error, rather than the evidence. The present of a few percent of S, Cl and K is also quite common in many organic samples, at least I met several times.
After all, I think that you should have more sufficient evidence to support your argument. The results of EDS could be trusted in many cases, but it failed to convince me this time.
Regards,
Charlie
Jose Luis Encinas wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hi, } } I am physicist and so my SEM-EDX application field is inorganic } material. I need to know whether a very small dried sample is blood. I } have not any device to biological sample preparation. I have obtained } the following EDX standardless semicuantitative analysis: } } Unknow sample My own blood sample } } C = 50.9 C = 47.4 } O = 39.3 O = 44.6 } Na= 0.5 Na= 1.2 } Si= 0.1 Si { 0.1 (presence) } P = 0.4 P = 0.2 } S = 1.8 S = 1.4 } Cl= 2.8 Cl= 2.7 } K = 3.2 K = 1.6 } Fe= 0.6 Fe= 0.4 } Cu { 0.1 (presence) Cu { 0.1 (presence) } } Are sufficient these data to conclude that the unknow sample is blood or } furthermore human blood?. If not, what may I to do?. } } - Thank you in advance -
Can anyone tell me of a supplier of thin sections of minerals and rocks for educational purposes? I am told that Carolina Biological is a possibility, and have requested their catalog, which is long in coming. Are there additional suppliers?
I'm not looking for labs that make thin sections from rocks and minerals that the customer supplies.
W. Riedel Scripps Institution of Oceanography UCSD La Jolla, CA 92093-0220
San Francisco Microscopical Society and California Association of Criminalists Trace Evidence Study Group
Joint Dinner Meeting Announcement October 30, 1997 Berkeley, California
Speaker: Brian J Ford Topic: Antony van Leeuwenhoek and the Single Lens Microscope
We are delighted to announce a very special evening featuring the noted English scientist and author, Brian J. Ford. Have you ever wondered how 17th and 18th century microscopists could prepare images of insects, cells, bacteria, and all manner of microscopic organisms and structures without SEMs, DIC, PCM, confocal microscopes, scanning tunneling microscopes and all those other marvelous inventions of the 20th century? How can a single lens microscope reveal structures that are even now difficult to study with sophisticated research microscopes? How indeed! Come hear some amazing answers to these questions.
Our program this month features a presentation on Antony van Leeuwenhoek (1632-1723) and single lens microscopy. Professor Ford is the author of countless books and papers and has done extensive research on the historical development of microscopy, including work with original, 17th century Leeuwenhoek instruments and samples from the archives of the Royal Microscopal Society in London. Many American microscopists know Professor Ford through his annual presentations at Inter/Micro in Chicago.
For further information on this meeting please see a special web page prepared for this special occasion: The URL is http://ourworld.compuserve.com/homepages/steve_shaffer/announce.htm - check it out NOW!
-- Submitted by Stephen A. Shaffer MicroDataware sshaffer-at-microdataware.com (business) steve_shaffer-at-compuserve.com (personal)
Dear list, there were a lot of questions conserning image processing software - that is one more. Does anyone can point as to a free-, share- or commercial software for crystal images processing (excluding CRISP - we know it well)? What we need other than different filters is to measure lattice distorsions and spots shifts on the large area.
The CI number for Methylene blue is 52015. The CI number for Azure B (sometimes called Azure I) is 52010. There is no CI number for Azure II since it is a 1:1 mixture of the the two stains above. Could you simply mix 52015 and 52010? I have not done this, simply because the Azure II worked fine, and I don't have time to fix things which already work reliably. I cannot think of any reason why mixing the two stains rather than buying the azure II would alter the situation. We buy our methylene blue and our Azure II from Sigma. I do not know if the percentage of active stain varies per gram between batches. This is something one needs to be aware of given the vast interactions between LM stains and embedding epoxies. I also used to buy very nice stains from Fisher (but someone stole our Fisher Catalog)! PS. I do not have any commercial interest in either of the companies mentioned. The price of these stains can vary enormously so it pays to check several sources. Just make sure that the company states the CI number of the product they are selling. Please note that the stain improves with age. Make a lot and let it sit in the dark at rt. Also please note that if the stained section is not rinsed with alcohol (75% - destained, and then with 100%), the result will not be as brilliant as it should be. Bye, Hildy
It is out of my field also, but I understand there is a chemical which, when applied to blood will cause it to floresce under UV light.... Perhaps someone else cam be more specific.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi,
I am physicist and so my SEM-EDX application field is inorganic material. I need to know whether a very small dried sample is blood. I have not any device to biological sample preparation. I have obtained the following EDX standardless semicuantitative analysis:
Unknow sample My own blood sample
C = 50.9 C = 47.4 O = 39.3 O = 44.6 Na= 0.5 Na= 1.2 Si= 0.1 Si { 0.1 (presence) P = 0.4 P = 0.2 S = 1.8 S = 1.4 Cl= 2.8 Cl= 2.7 K = 3.2 K = 1.6 Fe= 0.6 Fe= 0.4 Cu { 0.1 (presence) Cu { 0.1 (presence)
Are sufficient these data to conclude that the unknow sample is blood or furthermore human blood?. If not, what may I to do?.
I want to thank everyone that responded to my request for TEM labs. We are now in the process of wading through the myriad labs that were recommended. Hopefully, we will make a decision on which lab we will use within the next week. When we do, I'll be sure to contact each of you individually to let you know of our decision. Once again, thanks a bunch :-)
David Bell Millipore Corporation 80 Ashby Road Mailstop B2C Bedford, MA 01730
The largest perhaps is Ward's Scientific. They have many different sets of these. Pricy though.
Ward's Natural Science Establishment, Inc. 5100 West Henrietta Road Rochester, NK 14692-9012 1-800-962-2660 They may have a web page by now?
} Can anyone tell me of a supplier of thin sections of minerals and } rocks for educational purposes? I am told that Carolina Biological } is a possibility, and have requested their catalog, which is long } in coming. Are there additional suppliers? } } I'm not looking for labs that make thin sections from rocks and } minerals that the customer supplies. } } W. Riedel } Scripps Institution of Oceanography } UCSD } La Jolla, CA 92093-0220 ================================================ Michael L. Boucher Sr. mboucher-at-isd.net 13345 Foliage Avenue Apple Valley, MN 55124-5603 Ph 612-432-8836 ================================================
A few days ago, in response to an inquiry from Casey Lu about remote TEM operation, I posted the following:
Casey: } check out the URL: http://tpm.amc.anl.gov/MMC. } } Also, details about Remote Microscopy at the High Temperature } Materials Lab } at ORNL are given at } http://www.ms.ornl.gov/htmlhome/mauc/MAGRem.html.
It turns out that apparently the web page for the ORNL site does not display fully when addressed using Microsoft Internet Explorer. Netscape seems to work just fine. Also, earlier web pages up to htmlhome work OK on IE. We are checking to see what the problem might be. I will post when something is resolved, in case anyone else has encountered this problem.
Larry
Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
Electron microscopy specialist needed in spring 1998. Prepare ultra thin sections, and photogragh them using JEOL 1200EX. Darkroom, Adobe Illustrator/Photoshop.
Salary commensurate with experience.
Mail or fax resume and two letters of recommendation to:
Dr. Peter Sterling 123 Anatomy/Chemistry BLDG Department of Neuroscience University of Pennsylvania Philadelphia, PA 19104-6058
We have a Peltier cooled Critical Point Dryer (CPD750 - manufactured by EMSCOPE) that is having problems attaining a low enough temperature prior to flushing with carbon dioxide. The manual suggests a temperature below 10C - we can rarely get to below 12C and frequently only get to 14C. Ambient temperature at the moment is only around 20C. Has anyone any suggestions? As usual we have no specifications for the electronic circuit nor a circuit diagram.
Thanks
Ian
Ian Hallett HortResearch Mt Albert Research Centre Private Bag 92 169 Auckland, New Zealand Fax 64-9-815 4201 Telephone 64-9-849 3660 EMail ihallett-at-hort.cri.nz
Are we talking about Richardson's stain? (I don't have the reference to hand, but I could find it if anyone wants.) This is a 1:1 mixture of 1% methylene blue in 1% borax with 1% Azure II in water. It is my routine stain for thick sections and gives brilliant metachromatic results, even without an alcohol rinse.
Lesley Weston
On Tue, 14 Oct 1997, HILDEGARD CROWLEY wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Hi, } } The CI number for Methylene blue is 52015. } The CI number for Azure B (sometimes called Azure I) is 52010. } There is no CI number for Azure II since it is a 1:1 mixture of the the } two stains above. } Could you simply mix 52015 and 52010? I have not done this, simply } because the Azure II worked fine, and I don't have time to fix things } which already work reliably. I cannot think of any reason why mixing the two } stains rather than buying the azure II would alter the situation. } We buy our methylene blue and our Azure II from Sigma. I do not know if } the percentage of active stain varies per gram between batches. This is } something one } needs to be aware of given the vast interactions between LM stains and } embedding epoxies. I also used to buy very nice stains from Fisher (but } someone stole our Fisher Catalog)! PS. I do not have any commercial } interest in either of the companies mentioned. The price of these stains } can vary enormously so it pays to check several sources. Just make sure } that the company states the CI number of the product they are selling. } Please note that the stain improves with age. Make a lot and let it sit } in the dark at rt. Also please note that if the stained section is not } rinsed with alcohol (75% - destained, and then with 100%), the result } will not be as brilliant as it should be. } Bye, } Hildy }
Andrey L. Chuvilin wrote: -------------------------------------------------------------------. } } Dear list, } there were a lot of questions conserning image processing software - that } is one more. Does anyone can point as to a free-, share- or commercial } software for crystal images processing (excluding CRISP - we know it } well)? } What we need other than different filters is to measure lattice } distorsions and spots shifts on the large area.
Dear Andrey,
I know of 3 shareware programs that will measure and correct lattice distortions:
ICE: http://ncmi.bioch.bcm.tmc.edu/ftp.html http://condor.bcm.tmc.edu/3DEM/download.html ftp://ncmi.bioch.bcm.tmc.edu/pub/ICE/ which is a menu driven C-version of the Fortran-classic
MRC: http://www.EMBL-Heidelberg.DE/ExternalInfo/fuller/em_mrc_overview.html to obtain mail to: rac1-at-mrc-lmb.cam.ac.uk
You could also try EM (also a classic): to obtain mail to: hegerl-at-vms.biochem.mpg.de
A good source of information is the special edition of the Journal of Structural Biology, Vol. 116, No. 1, January/Februray 1996. It reviews most of the software generally used in the field.
Yours,
Philip
-- Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 86 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www_scem.csb.ki.se/pages/philip.html
Dear all, in our lab the question of carcinogenicity of propylenoxide arose. Where can get hard data about it, and which alternative can be recommended in Epon-embedding. Thank you all, Norbert
Ian, As long as the tempature is between 10-15C the CO2 will be a liquid. I use water from a sink to cool my CPD. Even in the summer the water is cool enough to get liquid CO2. I can't help you with your other problem.
-- Greg Rudomen Greg-at-umic.sunysb.edu S.U.N.Y. Stony Brook University Microscopy Imaging Center 516-444-3126
IAN HALLETT wrote:
} } We have a Peltier cooled Critical Point Dryer (CPD750 - manufactured } by EMSCOPE) that is having problems attaining a low enough } temperature prior to flushing with carbon dioxide. The manual } suggests a temperature below 10C - we can rarely get to below 12C and } frequently only get to 14C. Ambient temperature at the moment is } only around 20C. Has anyone any suggestions? As usual we have no } specifications for the electronic circuit nor a circuit diagram. } }
Ian, As long as the tempature is between 10-15C the CO2 will be a liquid. I use water from a sink to cool my CPD. Even in the summer the water is cool enough to get liquid CO2. I can't help you with your other problem.
-- Greg Rudomen Greg-at-umic.sunysb.edu S.U.N.Y. Stony Brook University Microscopy Imaging Center 516-444-3126
IAN HALLETT wrote:
} } We have a Peltier cooled Critical Point Dryer (CPD750 - manufactured } by EMSCOPE) that is having problems attaining a low enough } temperature prior to flushing with carbon dioxide. The manual } suggests a temperature below 10C - we can rarely get to below 12C and } frequently only get to 14C. Ambient temperature at the moment is } only around 20C. Has anyone any suggestions? As usual we have no } specifications for the electronic circuit nor a circuit diagram. } }
Ian, As long as the tempature is between 10-15C the CO2 will be a liquid. I use water from a sink to cool my CPD. Even in the summer the water is cool enough to get liquid CO2. I can't help you with your other problem.
-- Greg Rudomen Greg-at-umic.sunysb.edu S.U.N.Y. Stony Brook University Microscopy Imaging Center 516-444-3126
IAN HALLETT wrote:
} } We have a Peltier cooled Critical Point Dryer (CPD750 - manufactured } by EMSCOPE) that is having problems attaining a low enough } temperature prior to flushing with carbon dioxide. The manual } suggests a temperature below 10C - we can rarely get to below 12C and } frequently only get to 14C. Ambient temperature at the moment is } only around 20C. Has anyone any suggestions? As usual we have no } specifications for the electronic circuit nor a circuit diagram. } }
Ian, As long as the tempature is between 10-15C the CO2 will be a liquid. I use water from a sink to cool my CPD. Even in the summer the water is cool enough to get liquid CO2. I can't help you with your other problem.
-- Greg Rudomen Greg-at-umic.sunysb.edu S.U.N.Y. Stony Brook University Microscopy Imaging Center 516-444-3126
IAN HALLETT wrote:
} } We have a Peltier cooled Critical Point Dryer (CPD750 - manufactured } by EMSCOPE) that is having problems attaining a low enough } temperature prior to flushing with carbon dioxide. The manual } suggests a temperature below 10C - we can rarely get to below 12C and } frequently only get to 14C. Ambient temperature at the moment is } only around 20C. Has anyone any suggestions? As usual we have no } specifications for the electronic circuit nor a circuit diagram. } }
Ian, As long as the tempature is between 10-15C the CO2 will be a liquid. I use water from a sink to cool my CPD. Even in the summer the water is cool enough to get liquid CO2. I can't help you with your other problem.
-- Greg Rudomen Greg-at-umic.sunysb.edu S.U.N.Y. Stony Brook University Microscopy Imaging Center 516-444-3126
IAN HALLETT wrote:
} } We have a Peltier cooled Critical Point Dryer (CPD750 - manufactured } by EMSCOPE) that is having problems attaining a low enough } temperature prior to flushing with carbon dioxide. The manual } suggests a temperature below 10C - we can rarely get to below 12C and } frequently only get to 14C. Ambient temperature at the moment is } only around 20C. Has anyone any suggestions? As usual we have no } specifications for the electronic circuit nor a circuit diagram. } }
Ian, As long as the tempature is between 10-15C the CO2 will be a liquid. I use water from a sink to cool my CPD. Even in the summer the water is cool enough to get liquid CO2. I can't help you with your other problem.
-- Greg Rudomen Greg-at-umic.sunysb.edu S.U.N.Y. Stony Brook University Microscopy Imaging Center 516-444-3126
IAN HALLETT wrote:
} } We have a Peltier cooled Critical Point Dryer (CPD750 - manufactured } by EMSCOPE) that is having problems attaining a low enough } temperature prior to flushing with carbon dioxide. The manual } suggests a temperature below 10C - we can rarely get to below 12C and } frequently only get to 14C. Ambient temperature at the moment is } only around 20C. Has anyone any suggestions? As usual we have no } specifications for the electronic circuit nor a circuit diagram. } }
Ian, As long as the tempature is between 10-15C the CO2 will be a liquid. I use water from a sink to cool my CPD. Even in the summer the water is cool enough to get liquid CO2. I can't help you with your other problem.
-- Greg Rudomen Greg-at-umic.sunysb.edu S.U.N.Y. Stony Brook University Microscopy Imaging Center 516-444-3126
IAN HALLETT wrote:
} } We have a Peltier cooled Critical Point Dryer (CPD750 - manufactured } by EMSCOPE) that is having problems attaining a low enough } temperature prior to flushing with carbon dioxide. The manual } suggests a temperature below 10C - we can rarely get to below 12C and } frequently only get to 14C. Ambient temperature at the moment is } only around 20C. Has anyone any suggestions? As usual we have no } specifications for the electronic circuit nor a circuit diagram. } }
The Fall meeting of the Midwest Microscopy and Microanalysis Society will be held on Friday, November 7, 1997, at Eli Lilly & Co. in Greenfield, Indiana. The meeting is hosted by Jeffrey Horn of the Toxicology Research Laboratories at Lilly.
The program will begin at 8:30 a.m. and run until about 4:30 p.m. Lunch will be provided courtesy of Abbott Laboratories. In addition to podium presentations, there will be a tour of the Toxicology Facility at Eli Lilly.
The program includes a variety of topics - the proverbial "something for everyone" interested in microscopy and imaging. On the agenda are presentations about digital imaging, diagnostic electron microscopy, forensic electron microscopy, in situ labeling methods, and use of electron microscopy in the selection and development of pharmaceutical therapeutic candidates.
For more details about the program and meeting location, contact me by telephone at (847) 935-01014 or via E-mail at jane.a.fagerland-at-abbott.com.
Lilly is a secured facility, and pre-registration is requested to expedite entry and to allow us to plan for enough food for lunch. Please pre-register by contacting me at the phone or E-mail address above (E-mail is preferred).
Any supplier should be able to get you a MSDS (Material Safety Data Sheet) for it; ours came from Polyscience, 400 Valley Rd, Warrington, PA 18976, 215 343-6484. However WE DO NOT USE IT AT ALL AND HAVE NOT FOR OVER 15 YEARS. You can go straight from absolute ethanol into Eopn substitutes and Spurr as long as it is DRY. We use drying beads (molecular sieves) in all our 100% EtOH bottles. We keep only about 200-300 ml in a bottle, and when empty, we bake the beads. If there is silt, let it settle out and pipet off from the top. There has been a discussion of drying beads on this net recently.
On Wed, 15 Oct 1997, Deutschlaender, Norbert, Path. wrote:
} Date: Wed, 15 Oct 1997 10:07:00 +0200 } From: Deutschlaender, Norbert, Path. {DEUTSCHLAE-at-MSMPFEI.Hoechst.com} } To: "microscopy-at-sparc5.microscopy.c" {microscopy-at-sparc5.microscopy.com} } Subject: propylenoxide } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear all, } in our lab the question of carcinogenicity of propylenoxide arose. Where } can get hard data about it, and which alternative can be recommended in } Epon-embedding. } Thank you all, } Norbert }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
After years of always going from 100% ethanol to 100% propylene oxide before 1:1 epon:propylene oxide, I now go directly from 100% ethanol to 1:1 ethanol:epon and haven't seen any difference. Some resins (epon-araldite????) may require a propylene oxide intermediate but it doesn't seem to be required for epon only. } Dear all, } in our lab the question of carcinogenicity of propylenoxide arose. Where } can get hard data about it, and which alternative can be recommended in } Epon-embedding. } Thank you all, } Norbert
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
Dear Microscopists, We are using chicken embrio fibroblasts (CEF) for Golgi traffic study. We need specific Golgi staining from one side (cis- or trans-). We have tried to stain Golgi with several lectines - Helix Pomatia (HP) , Wheat Germ Agglutinin (WGA), Limax Flavus Agglutinin (LFA), but without any success. We used preembedding technices with lectines conjugated with horseradish peroxidase. HP simply did not stain CEF Golgi. WGA and LFA stained practically only endosomal compartment and rarely trans-Golgi network, but not trans-cisternes of Golgi. All these lectines work good in other cell types as NRK or RBL. I would like to ask does anybody know any specific lectin staining protocol for these cells. Any suggestions about lectin staining of cis- or trans- Golgi cisternes would be greatly appreciated.
At 03:59 PM 10/10/97 +0200, Philip Koeck wrote: } Does anybody have (good or bad) experience with any of the following } or other fast freezing devices: } Reichert KF80 } Leica CPC } Gentleman Jim (constructed by Alan Boyne) } Has anybody made a comparison?
I have had a long history of involvement with these devices, and believe that they are all good instruments which produce comparable results. That being said, I have sold the Gentleman Jim for more than 15 years, first with Ted Pella and now with Energy Beam Sciences, so my prejudices are obvious- the Gentleman Jim is much, much less expensive than the other instruments mentioned.
Best regards, steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
Donald P Robertson wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Could anyone suggest names of 3rd party vendors of LaB6 filaments } for an Hitachi H9000NAR. Thanks in advance for any advice. } Donald Robertson
Ladd Research supplies LaB6 filaments for Hitachi scopes under the catalog # 63070. If you wish a quote just contact us.
John Arnott Ladd Research ladres-at-worldnet.att.net tel 1-800-451-3406 fax 1-802-878-8074
I'm looking for a digital camera for an optical microscope that I can control from a program, such as a Visual Basic program, for setting up an automated imaging system. Anyone out there doing this type of thing?
} We have a Peltier cooled Critical Point Dryer (CPD750 - manufactured } by EMSCOPE) that is having problems attaining a low enough } temperature prior to flushing with carbon dioxide. The manual } suggests a temperature below 10C - we can rarely get to below 12C and } frequently only get to 14C. Ambient temperature at the moment is } only around 20C. Has anyone any suggestions? As usual we have no } specifications for the electronic circuit nor a circuit diagram. } Dear Ian, Peltier units can go bad. If you can measure the current they draw now and compare that with what they are supposed to draw, you can ascertain whether the unit is working as specified. Generally, the units are in the form of an array of individual devices. I don't re- member if these are series or parallel, but if one or more has shorted (series) or failed open (parallel), the unit will draw more/less current than it should. Since you have neither specs nor diagrams, this info may not be useful, but perhaps someone else can let you know what cur- rent a properly-working device draws. Good luck. Yours, Bill Tivol
I have also avoided propylene oxide for years for safety reasons. (Although I was warned of its flammability/volatility).
Alternatives: Usually I work with Spurr's resin (OK also carcinogenic but more easy to contain) and when dehydrating in acetone no transition solvent is needed. For ethanol, on the other hand, a transition solvent can be very useful in my experience with samples from pig skin to cell cultures. What to use? I have converted to tert-butyl glycidyl ether (t-BGE), available from Aldrich Chemical, Milwaukee, WI USA (cat. #25,171-2). This was recommended in a 1995 Biomaterials Workshop lecture by Hendrik K. Koerten in San Francisco, CA, sponsored by the Society for Biomaterials. It has been particularly useful to me in that it does not dissolve polymer materials/implants as readily as propylene oxide. Another name for this chemical is Butyl-2,3-epoxypropylether.
I have also used t-BGE with Araldite and EmBed 812 (an Epon-clone from Electron Microscopy Sciences), and it worked well.
Another good, or better, choice with the eponates is HPMA (hydroxypropyl methacrylate) which also is more kind to materials (cell culture plates) than propylene oxide.
I believe both HPMA and t-BGE are less flammable/carcinogenic than propylene oxide.
Karen
deutschlae-at-msmpfei.hoechst.com wrote: } } Dear all, } in our lab the question of carcinogenicity of propylenoxide arose. Where } can get hard data about it, and which alternative can be recommended in } Epon-embedding. } Thank you all, } Norbert
-- Karen Zaruba kszaruba-at-mmm.com 3M Company, 3M Center Bldg. 270-1S-01 St. Paul, MN 55144 "The opinions stated above are my own, not necessarily 3M's"
Title: Biological Science Technician (Plants) Lab: USDA-ARS, Salinas, CA
The USDA-ARS is seeking a biological science technician (plants) (GS-0404-6/7/8/9) for the Crop Improvement and Protection Research Unit in Salinas, CA. The incumbent will share responsibilities in electron microscopy of plant virus infections for ultrastructural characteristics. The incumbent will also assist in research involving molecular, serological, and biological studies of several plant viruses infecting sugarbeet and vegetables. Candidate must have a knowledge of electron microscopy, plant virology, and knowledge of microbiological techniques. Must be a U.S. citizen. Bachelors degree is desirable. Salary is commensurate with experience ($22,805-$40,300 per annum). For information regarding research program contact Gail C. Wisler or James E. Duffus (408)755-2835. For information regarding application procedures/forms contact Elsa Chavez at (408)755-2800. Applications must be postmarked by Nov 3, 1997. The USDA is an equal opportunity employer.
Gail C. Wisler USDA-ARS 1636 E. Alisal St. Salinas, CA 93905 (408)755-2835 FAX (408)755-2814 e-mail: gwisler-at- ASRR.ARSUSDA.gov
John R Reffner wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } I'm looking for a digital camera for an optical microscope that I can control } from a program, such as a Visual Basic program, for setting up an automated } imaging system. Anyone out there doing this type of thing?Hi, John,
I believe most of the digital cameras have this capability. In the past, we have had some experience with those from Kodak, Leaf, and Pixera. Also, Polaroid just announced a new digital camera. I would also check to see what Optronics and Dage-MTI are carrying. RE: outlets - call your local system integrator or the manufacturer's directly. (The Kodak Division you need is the one in New Haven; the Polaroid Group is under Jim Landrigan). If you have trouble finding a local source, please email me for manufacturers' contacts.
Best regards,
Barbara Foster President Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108 PH: (413)746-6931 FX: (413)746-9311 email:mme-at-map.com --------------------------------------------------------------------------------------------------------------------------------- ********** Microscopy/Microscopy Education ********** America’s First National Consortium of Microscopy Experts Specializing in Customized, On-site Training in all areas of Microscopy, Sample Prep, and Image Analysis
Title: Biological Science Technician (Plants) Lab: USDA-ARS, Salinas, CA
The USDA-ARS is seeking a biological science technician (plants) (GS-0404-6/7/8/9) for the Crop Improvement and Protection Research Unit in Salinas, CA. The incumbent will share responsibilities in electron microscopy of plant virus infections for ultrastructural characteristics. The incumbent will also assist in research involving molecular, serological, and biological studies of several plant viruses infecting sugarbeet and vegetables. Candidate must have a knowledge of electron microscopy, plant virology, and knowledge of microbiological techniques. Must be a U.S. citizen. Bachelors degree is desirable. Salary is commensurate with experience ($22,805-$40,300 per annum). For information regarding research program contact Gail C. Wisler or James E. Duffus (408)755-2835. For information regarding application procedures/forms contact Elsa Chavez at (408)755-2800. Applications must be postmarked by Nov 3, 1997. The USDA is an equal opportunity employer.
Gail C. Wisler USDA-ARS 1636 E. Alisal St. Salinas, CA 93905 (408)755-2835 FAX (408)755-2814 e-mail: gwisler-at- ASRR.ARSUSDA.gov
We have attempted to immuno-label one micron sections cut from LR White with primary antibody followed by TRITC or FITC conjugated secondaries. The sections are dried onto glass slides at 60C (the same temperature at which the blocks were cured), the immunolabel procedure performed, and a cover glass mounted with aquamount (UV clear). These experiments were completely unsuccessful in the fluorescent microscope, even though the same immunolabeling protocol works well in the EM with the antibodies and immuno-gold secondaries applied to the surface of ultrathin sections. I realize there are better ways to label tissue with antibody in preparation for fluorescence microscopy, but we are surprised by the negative result in these experiments. Does anyone have an explanation for the failure? Am I missing something simple? Is it because there is no penetration of primary and secondary into the tissue sections, therefore not enough bound TRITC or FITC to detect? It would be nice if this technique worked so that we could easily test the ability of our primaries to bind specifically to LR White embedded tissue.
Many thanks,
Doug Keene Laboratory for Ultrastructural Research Portland Shrine Research Unit ---------------------- Doug Keene DRK-at-shcc.org
OOPS! I share my husband's mailbox and erased the new address for Alwyn Eades. Where did he go? To avoid multiple responses, please limit reply to those who reside in Chicago, Ill. or immediate vic.
} I heard about a new x-ray microscopy technique that can image cellular } processes in real-time. Have you heard about this? } } } Bob Clark } } Hi ! Interesting idea, but it doesn=B4st seem very likely that it would be possible. According to a simple calculation we made, imaging of a cell would kill it within a few milliseconds, using x-rays. If someone has other information it would be interesting to share it !
bengt Bengt Stocklassa , Managing Director Cox Analytical Systems AB | Phone: +46 31 7725300 House of Innovations, CTH | Fax: +46 31 7725600 412 96 - Gothenburg, SWEDEN | E-mail: bengt-at-xco.se
A Research Fellowship is available from 1 January 1998 for a fixed period until 30 June 1999. The apointee will work on an electron microscope study of high-purity iron alloys to determine the mechanism of formation of coarse side-plate structures from grain boundary polygonal ferrite, and the effect of alloying elements on this transition, which would provide a means, via suppression of this reaction, to produce interlocking acicular ferrite microstructures from intragranular nucleation.
Applicants should have, or be submitting for, a PhD in Metallurgy, Materials Science or a related discipline. Experience in electron microscopy would be advantageous.
Salary will be on the scale for Research Staff Grade 1A within the range stlg15,159 - stlg18,494 p.a. according to qualifications and relevant experience.
Application forms and further particulars may be obtained from Professor D V Edmonds, Department of Materials, University of Leeds, Leeds, LS2 9JT, tel +44 0113 233 2341/8, fax +44 0113 233 3284, email d.v.edmonds-at-leeds.ac.uk
Closing date for applications 7 November 1997.
The University of Leeds promotes an Equal Opportunities Policy. _____________________________ Dr. Rik Brydson, University Research Fellow, Electron Optical Unit, Department of Materials, School of Process, Environmental and Materials Engineering University of Leeds, Leeds LS2 9JT, U.K.
Dear all, 10/16/97 unfortunately I don=B4t know at which point I=B4m entering = considerations and discussions (due to several failings of our Server). Following the discussion as far as I got the postings to my e-mail-box = (original question by Norbert Deutschlaender, Re Malcolm Haswell, Re = Sara E.Miller, Re Thomas E.Phillips , Re Karen Zaruba) for me there are = several points worth to consider: i) propyleneoxide (PO): why, when, which substitutes possible I was using PO for specimen preparation during my studies in Zoology = about 20 years ago. I was frustrated because of the odious smelling of = PO-vapors and its solving reactivity to safety wear like gloves and = plastic labware. When I got then for the first time a MSDS (material = Safety Data Sheet) on PO and searched for the chemical nature of the = substance I got aware of its properties as a carcinogen, I knew from the = beginning that it is a highly flammable solvent and there were severe = cautions in handling the substance....The main reason(s) for using this = solvent PO in my opinion was/were the high rate of volatilization at = room temperature (20 degr.C), which is, according to MSDS about 588 hPa = (at 33 degr.C: 980 hPa, compare data for Aceton and Acetonitrile, see = below): so in fact that "intermedium" (which as such should aid in the = miscibility of the hydrophilic alcoholic solutions with the hydrophobic = nature of the embedding resin, sometimes also said to be a = "clearing[??]agent") after several changes evaporated as a pure = hydrophobic solvent out from the resin. To produce "no shock treatment" = on the tissues which eventually could lead to severe ultrastructural = changes in morphological terms, that exchange between hydrophilic and = hydrophobic phases usually was/is done by including several changes of = the tissues=20 in relations of EtOH:PO e.g. 3:1, 1:1, 1:3, then pure PO e.g. 3 changes, then pure PO:Epon(-oxide) e.g. 3:1, 1:1, 1:3 then pure resin e.g. several changes to get rid of traces of the = solvent PO.
ii) It turned out many years ago that at least EPON (as a hydrophobic, = non water-miscible substance) only up to 96% would be hydrophobic. It = was said to have a hydrophilic phase at/up to a concentration of 4%, = namely "aquon"-phase. This allowed for an embedding only via 100%, = absolute water-free EtOH, provided several "protected" (against = watervapor-uptake) changes in pure such EtOH as well as several changes = in mixtures of abs.abs. EtOH / Epon were performed. In addition the = "infiltrating" time of each such step had to be prolonged (at least = doubled). I remember the days when the "new" hydrophilic resins like LR White or = the Lowicryls were not "born" yet and people tried to overcome = interactions of organic solvents with tissue antigens at the beginning = of ultrastructural localization of antigens by Antibodies.
iii) I don=B4t have experience with t-BGE (as mentioned by Karen = Zaruba). Karen mentions also the possibility to dehydrate by Acetone = rather than Ethanol. Yes, you don=B4t need an intermedium like PO, = provided the several changes of tissues as mentioned in i). As a = reference (maybe out of many possible/present references) I can quote: = BULLOCK et al (Eds, 1989): Techniques in diagnostic Pathology, Vol. 1, = p. 1 ff (Academic Press, ISBN: 0-12-681911-4) where it is stated that = "Acetone (is) slightly superior than Ethanol as dehydrating agent for = better preservation of Immunoreactivity/Antigenicity in processing Bone = Marrow Aspirates in Plastic". Acetone has a lower rate of volatilization = at room temperature (20 degr.C), which is, according to MSDS about only = 233 hPa (at 33 degr.C: no data available, compare data for Acetonitrile, = see below and PO, see above).
For several reasons (mainly for safety reasons) I use now for a long = time (} 10 years) a substitute for PO, namely ACETONITRILE (syn.: = Methylcyanide) for dehydration, or as an Intermedium, respectively. When = I saw the 2 page publication (CAVE: text p 38: : ACETONITRILE: = Substitute for Propylene Oxide in Tissue Processing for Transmission = Electron Microscopy ; CAVE: Illustrations erroneously are printed on = page 41 instead page 39) by:=20 TARNOWSKI Betty I., & SCHONBAUM Greg R. in the Proceedings of EMSA, = 42nd Ann. Meeting ( 1984) / Detroit I learned a lot of new aspects.
I do not know wether there is an earlier paper on that subject or an = other author(s) to be named for publishing first that substitution = method (please apologize).
Product info: ACETONITRILE: 2-cyanopropane-2-ol, (C 2 H 3 N), MW: 41.05 = g/mol, 1 liter =3D 0.78 kg; melting point: -46 degr.C., boiling point: 81 = degr.C, flash point: 5 degr.C, 100% miscible with water, free from = halogens; e.g. from SIGMA Order# A 3396 (Disclaimer: I have no interest in = SIGMA=B4s..... nor am I affiliated with Sigma....)
Acetonitrile (AN) has -compared to Acetone and PO- the lowest rate of = volatilization at room temperature (20 degr.C), which is, according to = MSDS about only 97 hPa (at 33 degr.C: no data available, compare data = for Acetone, and PO, see above). It is indeed a toxic substance = (methyl-cyanide): due to the reduced vapor-pressure you will have a = tremendous decrease in the possibility of inhaling the toxic vapors when = working with it. Additionally, AN is not classified as carcinogen, so if = you work carefully in a well-ventilated area, you wouldn=B4t need a = filter-mask or even a fume-cupboard (in practical work). At least you = should not eat or drink the solvent! The substance is fully (100%) miscible with water, so, in fact, you = could dehydrate and intermediate, infiltrate by/with only one solvent.=20 I haven=B4t tried a complete dehydration with increasing concentrations = of only Acetonitrile but I am convinced such would work (provided you = can prevent tissues from "rehydration" by hygroscopic saturation of the = solvent via water-vapors or high humidity).=20 As the substance at all has to be classified as toxic, flammable and = vapors to be a health-risk, I=B4m not using it as the only dehydrating = agent.
After I checked that substitution possibility in comparing control = specimen processings (i.e. PO vs. AN) and got aware of that no = alterations in ultrastructural morphology could be detected between PO = and AN-specimens, from 1985 on I have completely changed to the = following schedule for (automatically) processing my diagnostic tissue = samples (standardized protocol, approx. 1500-1600 blocks/year): increasing concentrations of alcohols (as usual), - 4 x EtOH absolutely waterfree (maybe there will be rehydration of = about=20 0.5-1%, when opening bottle and handling: this is no problem at all) (times: each 5/10/10/10 mins) - 3 x pure ACETONITRILE (each 5, 10, 15 mins) - infiltration by mixture=20 EPOXIDE RESIN : ACETONITRILE =3D 1:1 (at least 45 mins) (GLYCIDETHER 100,=20 formerly SERVA/now: BIOPRODUCTS) then - 3 x pure EPOXIDE RESIN (at least until tissue blocks have sunk down = to the bottom of the "infiltrating mold" or infiltrating glass).=20 Eventually I recommend the use of a bulb lamp (e.g. 60 W) to be = positioned approx. 10 cm above the infiltrating mold, at least of the = first pure resin step to aid in evaporation of Acetonitrile remnants in = the tissue/resin mixture. In doing that way, up to now no problems in infiltration- and = polymerization quality occured (except when wet tissue blocks exceeded = dimensions L/W/H of 4x5x1.5 mm, where one should increase duration of = the processing steps).
The suggestion of Sara Miller (or her method) to use molecular sieves = (drying beads) in all of their 100% EtOH bottles I have to mention that = I got serious problems with a following damage of the cutting edge of = our diamond knives when sectioning the blocks (due to silt produced from = the molecular sieve beads, which were infiltrating the tissues and = therefore embedded too). But: maybe this happened due to pouring of 100% = EtOH from the bottle rather than pipetting off from the top of the = solution.
At the end: answering the question of Malcolm Haswell "opinion about which is nastier: Spurr=B4s resin with alcohol or Epon with propylene oxide" I have to "confess": if you have to choose between "devil" and "satanas" you should try to reach the "purgatory" that means: Try to reduce toxicity, handling and healthy hazards, = maintain safety implications, be aware of "the devil and satanas".
Best regards to all Wolfgang MUSS EM-Lab of Dept. Pathol. LKA A-5020 SALZBURG, AUSTRIA/Europe
Hi, 10/16/97 unfortunately I am not aware of a commercial source of Uranyl formate, = which I think has been used as negative staining agent in former years. As an indication I found a fax notice from 1984 that a company in = Germany, namely PAESEL Chemicals (I wonder if they are still alive) = which offered 5 g of that substance at a price of US$ 53.-. Now I have had a look in my files and a phone call: the company is
PAESEL GmbH&Co=20 Moselstrasse 2 B D-63 452 HANAU Federal Republic of Germany phone: ++49++6181-18-700 Fax: ++49++6181-18-7070=20
They told me to have listed Uranyl-formate in quantities of 5 gs, no = price-information could be given at the moment.
Note added: If you still have to produce your uranyl-formate by yourself, you should = have a look on 2 original publications I found, dealing with the = fabrication of Uranylformate:=20
LEBERMANN R (1965): Use of Uranylformate as a Negative Stain, J. Molecul. Biol. 13, p.606 ff
and an alternative preparation method:
BRADLEY D.E. (1965) in J. Gen. Microbiology 38, p. 395 ff
Best wishes and luck to you
Wolfgang MUSS Dept. Pathology LKA, EM-Lab A-5020 SALZBURG, Austria/Europe.
Hi, 10/16/97 unfortunately I am not aware of a commercial source of Uranyl formate, = which I think has been used as negative staining agent in former years. As an indication I found a fax notice from 1984 that a company in = Germany, namely PAESEL Chemicals (I wonder if they are still alive) = which offered 5 g of that substance at a price of US$ 53.-. Now I have had a look in my files and a phone call: the company is
PAESEL GmbH&Co=20 Moselstrasse 2 B D-63 452 HANAU Federal Republic of Germany phone: ++49++6181-18-700 Fax: ++49++6181-18-7070=20
They told me to have listed Uranyl-formate=20
ADDED NEW: Order# 3 36 234
in quantities of 5 gs, no price-information could be given at the = moment.
Note added: If you still have to produce your uranyl-formate by yourself, you should = have a look on 2 original publications I found, dealing with the = fabrication of Uranylformate:=20
LEBERMANN R (1965): Use of Uranylformate as a Negative Stain, J. Molecul. Biol. 13, p.606 ff
and an alternative preparation method:
BRADLEY D.E. (1965) in J. Gen. Microbiology 38, p. 395 ff
Best wishes and luck to you
Wolfgang MUSS Dept. Pathology LKA, EM-Lab A-5020 SALZBURG, Austria/Europe.
Greetings, Doug Keene wrote: } } We have attempted to immuno-label one micron sections cut } from LR White with primary antibody followed by TRITC or } FITC conjugated secondaries. ... } These experiments were completely unsuccessful in } the fluorescent microscope, even though the same } immunolabeling protocol works well in the EM with the } antibodies and immuno-gold secondaries applied to } the surface of ultrathin sections. ....
Keep in mind that the propoprtion of surface area to volume is much higher for a 60 nm ultra section then for a 1 um semithin section. For LR White, if the ab can make it to antigens as deep as 15 nm (I am just guessing at the depth here), then it will find the antigen in half the ultrathin section volume but only in 3% of the semithin section. You may be able to improve your signal at the light level with one of the antigen retrival methods that are around. I have never done these myself, but I have heard that they will work. Alternatively, you can embed your samples in butyl-methyl methacrylate, which is extractable after sectioning and thus gives great access for your ab throughout the section volume. Of course, this resin is not so great in the em (although useable) and so this path would mean that you would have different preps for light and em work (although I believe that LR white is a methacrylate based resin). Hope this helps, Tobias Baskin
Propylene oxide can be omitted from an embedding protocol.
However! it is important to realize that 1)alcohol interferes with polymerization 2)acetone also interferes with polymerization 3)a mixture of PO and 812 is much less viscous than a mixture of 812 and acetone and alcohol 3) PO is actually a monoexpoxide and if remnants are left in the tissue it will become incorporated into the block. The critical issue in many infiltration steps is the viscosity of the monomers. Skin may require PO, liver can do nicely without it.
We have abondoned PO a long time ago. We use acetone instead. However, if the blocks are particularly large or difficult then we go to PO temporarily to insure adequate infiltration. We make absolutely sure that no acetone or alcohol is left in the mixture. This means changing 812 more often, and changing to clean vials after the tissue has been in the first undiluted 812 for one hour.
Dear list We are progressing nicely with digital imaging and printing from our microscopes. We have several networked printers that people are using. I now need to be able to charge accordingly for the printouts that people do. I am using WIN95 networking and NT4.0 server (and Novell 3.12 but want to stay away from that for network printing if I can). Is anyone using server software which will record which printer was used, who used it and how many printouts they did? NTserver event manager is sort of working but seems always to say 0 pages were printed. If anyone has gone down this route I would appreciate some comments.
Raining again in Manchester!
Chris
Chris Gilpin Biological Sciences Electron Microscope Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 161 275 5170 fax +44 161 275 5171 http://www.biomed.man.ac.uk/biology/emunit/emhome.html
I routinly test 1 micron LR White sections on glass before going to the trouble of Immuno EM. I usually use peroxidase ABC which seems to enhance the signal enough to be visualized. However, the fluorescent signal is very weak unless you start stacking antibodies which can create more backround. So you need a very good optimized scope to be able to see the signal and your faint signal may be obscured by the backround signal of the aquamount which has higher fluorescence than some others. Prolong from Molecular Probes is very quiet and enhances Texas Red. Or my favorite for quiet and quick is: 70% Glycerol, 25% .5M Tris pH 9, 5% n-Propylgallate, heat to mix then pH to 7.4 and store in the fridge. For use: 1 drop, swirl, drain most of it off then coverslip. There should be barely enough to cover the coverslip.
Good Luck Doug
Robert Underwood Morphology Core Dermatology U of Wash. Seattle, WA On Thu, 16 Oct 1997, Doug Keene wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Fellow Microscopists: } } We have attempted to immuno-label one micron sections cut } from LR White with primary antibody followed by TRITC or } FITC conjugated secondaries. The sections are } dried onto glass slides at 60C (the same temperature at } which the blocks were cured), the immunolabel procedure } performed, and a cover glass mounted with aquamount (UV } clear). These experiments were completely unsuccessful in } the fluorescent microscope, even though the same } immunolabeling protocol works well in the EM with the } antibodies and immuno-gold secondaries applied to } the surface of ultrathin sections. I realize there are } better ways to label tissue with antibody in preparation } for fluorescence microscopy, but we are surprised by the } negative result in these experiments. Does anyone have an } explanation for the failure? Am I missing something } simple? Is it because there is no penetration of primary } and secondary into the tissue sections, therefore not } enough bound TRITC or FITC to detect? It would be nice if } this technique worked so that we could easily test the } ability of our primaries to bind specifically to LR White } embedded tissue. } } Many thanks, } } Doug Keene } Laboratory for Ultrastructural Research } Portland Shrine Research Unit } ---------------------- } Doug Keene } DRK-at-shcc.org } } }
Our lab is in the process preparing swatches of 50/50 ploycotton fabric contaminated with bacteria for SEM. The swatches will be processed using the fairly standard method, for this lab, of aldehyde, osmium, and ethanol.
HAS ANYONE HAD POSITIVE OR NEGATIVE RESULTS USING HMDS RATHER THAN CPD ON TEXTILES/CLOTH?
At 10:39 AM 10/16/97 GMT+1200, Ritchie Sims wrote: } Does anyone know of a commercially-available embedding resin kit } using methyl and/or butyl acrylate and/or methacrylate? } Preferably email and/or website contact.
There is a methyl methacrylate embedding kit manufactured by Heraeus Kulzer in Germany and sold under the name "Technovit 9100." Unfortunately, the activator, Percodox 16, an organic peroxide manufactured by Akzo Chemical, is classified as an explosive. This effectively prevents the kits from being shipped by air. Kulzer is working on a replacement protocol using a different activator but, in the meantime, the kits are not available.
Disclaimer: Energy Beam Sciences sells the Technovit GMA and (we hope) MMA kits in the United States.
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
Chris: Windows NT 4.0 should be able to to maintain a log for printers. We use NT 4.0 Workstation to record the user, the printer used, the name of the document printed, and the number of pages. We charge for prints from our TEK Phaser 440. This is how to set it up to log. 1. Start} settings} printers 2. double click the name of the printer you want to audit 3. Select Printer from the menu} properties} security} auditing 4. Select Add, select the name of the group you want to audit (I just select "Everyone"), select Add, then select OK, then select OK in the Properties page. 5. In the Printers page, select File} Server properties} Advanced} Log Spooler information events, then select OK. 6. Then restart the computer. 7. In the Event viewer in Administrative Tools it will record all print events in the System Event log.
I have found NT to be a great operating system once you figure out what you want to do. Hope this will help. Stanley L. Flegler, Assistant Director Center for Electron Optics Michigan State University US flegler-at-pilot.msu.edu
The UCF/Cirent Materials Characterization Facility has an immediate opening for an ion probe engineer with expertise in the operation and maintenance of SIMS equipment. Expertise in the operation and maintenance of an RBS accelerator is also a plus. The MCF at the University of Central Florida in Orlando is a 5400 sq. ft. facility which includes 2 FEG SEM's (Hitachi, JEOL), 2 TEM's (Philips 400T, JEOL 2000FX), a PHI Auger, 3 SIMS (PHI, Riber, Cameca IMS 3f), a JEOL 733 microprobe and an 1.7MeV RBS accelerator. The engineer will be responsible for the daily operation and maintenance of SIMS equipment. It is also expected that the engineer will work closely with MCF faculty, staff, students and industrial affiliates. UCF is close to Cirent Semiconductor (Lucent Technologies), Lockheed Martin, NASA Kennedy Space Center, Westinghouse, Universal Studios, Walt Disney World, Harris Corp., Pratt and Whitney, and others. Please send a resume and a list of references to: Dr. L.A. Giannuzzi, Director, UCF/Cirent Materials Characterization Facility, Mechanical Materials & Aerospace Engineering, PO Box 162450, 4000 Central Florida Blvd., Orlando, FL 32816-2450 or send an email Word attachment to: lag-at-pegasus.cc.ucf.edu. UCF is an equal opportunity affirmative action employer.
************************************************************************* Lucille A. Giannuzzi, Ph.D.
Assistant Professor Dept. of Mechanical, Materials, and Aerospace Eng.
Director, Cirent/UCF Materials Characterization Facility President, Florida Microscopy Society (a local affiliate of MSA)
University of Central Florida phone (407) 823-5770 PO Box 162450 fax (407) 823-0208 4000 Central Florida Blvd. email lag-at-pegasus.cc.ucf.edu Orlando, FL 32816-2450 USA ----------------------------------------------------------------------- "Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain *************************************************************************
We have an LKB 2208 Multiplate wax heater for mounting plastic boats onto glass knives. It seems to be running a few degrees cool as the wax solidifies too soon upon contacting the knife with the boat. Does anyone know of a way to increase the temperature?
Bob
Dr. Robert R. Wise Department of Biology University of Wisconsin-Oshkosh Oshkosh, WI 54901
(920) 424-3404 tel (920) 424-1101 fax wise-at-uwosh.edu
Deutschlaender, Norbert, Path. wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Dear all, } in our lab the question of carcinogenicity of propylenoxide arose. Where } can get hard data about it, and which alternative can be recommended in } Epon-embedding. } Thank you all, } Norbert
Norbet,
Propylene oxide is an NTP anticipated carcinogen and is an IARC category 2B (probable cacinogen. For further information contact National Toxicology Program Office at 919-541-0530 or the WHO Publications Center at 518-436-9686. Acetone and ethanol are viable alternatives to Propylene oxide. Ethanol should be used cautiously because it may inhibit epoxy polymerization.
Hope this helps,
Charles Duvic Chief Chemist Ladd Research tel 1-800-451-3406 fax 1-802-878-8074
Chris: I forgot one part of step 4. Step 4 should read: Select Add, select the name of the group you want to audit (I just select "Everyone"), select Add, then check Print Success, Failure, etc. as needed in the Printer Auditing page, then select OK, then select OK in the Properties page. Sorry for the confusion. Stanley L. Flegler, Assistant Director Center for Electron Optics Michigan State University US flegler-at-pilot.msu.edu
We have a couple you could use, but I'd need more info from you before I could recommend one; such as: when you say you are looking for digital, do you mean the image output, or simply a digital control?, color or monochrome?, what level of support of the camera control do you need?, etc.
Please feel free to contact me using the information below.
****************************** Jim Haley Applications Engineer I-CUBE 2411 Crofton Lane, Suite 14A Crofton, MD 21114 voice: (301) 858-0505 fax: (301) 858-0615 web site: http://www.i-cubeinc.com e-mail: haley-at-i-cubeinc.com ******************************
John R Reffner wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I'm looking for a digital camera for an optical microscope that I can control } from a program, such as a Visual Basic program, for setting up an automated } imaging system. Anyone out there doing this type of thing?
I read an article on X-ray microscopy in the Jan/Feb. 1996 issue =
of Biophotonics International (by Laurin Publishing), pg. 58. =
The article highlights work by researchers (Dr. Cathie Magowan) =
at the Ernest Orlando Lawrence Berkeley National Laboratory, =
studying interactions between malarial parasites and host cells. =
They studied living [I think] parasites in red blood cells over =
48 hours, in aqueous medium. The stated resolution of the system =
was 60 nm in X-ray mode; the microscope could be switched between =
visible light and X-ray modes on the same sample.
If anyone knows more about this I'd be interested out of =
curiosity. Being a biologist I don't really want all the gory =
details; I just want to know practicle applications, =
availability, cost, etc. of the instrumentation.
Thanks, Karen
bengt-at-mail.coxsys.se wrote: } =
} } =
} } I heard about a new x-ray microscopy technique that can image cellular=
} } processes in real-time. Have you heard about this? } } } } } } Bob Clark } } } } Hi ! } Interesting idea, but it doesn=B4st seem very likely that it would be } possible. According to a simple calculation we made, imaging of a cell } would kill it within a few milliseconds, using x-rays. If someone has o= ther } information it would be interesting to share it ! } =
} bengt } Bengt Stocklassa , Managing Director } Cox Analytical Systems AB | Phone: +46 31 7725300 } House of Innovations, CTH | Fax: +46 31 7725600 } 412 96 - Gothenburg, SWEDEN | E-mail: bengt-at-xco.se
-- =
Karen Zaruba =
kszaruba-at-mmm.com 3M Company, 3M Center Bldg. 270-1S-01 St. Paul, MN 55144 "The opinions stated above are my own, not necessarily 3M's"
I'd like to de-embed some LX112 embedded plant leaf tissue so that I could examine it in the SEM but, never having done this before, I'm looking for advice, tips, references, protocols etc. Also, is it possible to de-embed thin sections already stained (uranyl acetate, lead citrate) and examined in the TEM? (Assuming I could get them off the grid....)
} Our lab is in the process preparing swatches of 50/50 ploycotton } fabric contaminated with bacteria for SEM. The swatches will be } processed using the fairly standard method, for this lab, of aldehyde, } osmium, and ethanol. } } HAS ANYONE HAD POSITIVE OR NEGATIVE RESULTS USING HMDS RATHER THAN } CPD ON TEXTILES/CLOTH? } } Thanks, Lloyd.
I haven't tried HMDS on 50/50 polycotton, but I have used it with good success on cotton fibers and polycarbonate membranes, as well as other compounds. I would expect it to work fine, but it might be better if you specified the polymer in the blend. (Have you contacted the vendor of the HMDS?)
An excellent source for Image Analysis/ Imaging techniques for any topic is Dr. John Russ's book "The Image Processing Handbook" which is available from CRC press.
David Bell Millipore Corporation Mailstop B2C 80 Ashby Road Bedford, MA 01730
} I am trying to to find a reference for the Epon-Spurr resin recipe that is } used for microwave fixation. } } I have listed here that the resin can be poylmerized at 70 degree C for } 24 hours but I ned a reference that says I can do that.. } } Any suggestions or does anyone have the reference } } Thanks } } Eric } Fred Hutchinson Cancer Research Center } }
I have tried labelling 1 micron sections with lectins conjugated with TRITC and they worked exceptionally well.... that is the lectin was labelled with the TRITC tag..
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Tomorrow evening there will be a meeting in Chicago of the State Microscopical Society of IL (SMSI). The speaker is Riccardo Levi-Setti, director of the Enrico Fermi Institute at the Univ. of Chicago and he will speak about his work on trilobites. If you are interested in details, let me know for I will be going.
==================================================================== FALL MEETING OF THE APPALACHIAN REGIONAL MICROSCOPY SOCIETY (AReMS) October 23-24, 1997
Host: Clinch Valley College of the University of Virginia, Wise, VA
Registration: Oct. 23, Noon until 5:00 pm, Holiday Inn, Norton, VA (For additional information and pre-registration, see the bottom of this notice.)
Thursday, October 23, 1997, WORKSHOPS:
Workshop 1. 2:00 - 3:00 "HTML and Web Design" Led by Dr. Herb Brown, Dir. Instructional Technology, CVC. Will cover the basics of web page creation, using the Netscape Navigator Gold editor. Hypertext Markup Language (HTML) will be explained and interpreted. Participants will create their own basic web page with text, graphics, and hypertext links. Good web page design techniques will be fostered. Participants will only need basic computer skills.
Workshop 2. 2:00 - 3:00 "E-mail and networked communications" Led by Mr. Alex Edwards, Assoc. Professor of Computer Science, CVC.
Workshop 3. 2:00 - 3:00 "An Introduction to the SEM" Led by Dr. Stan Kunigelis, Assoc. Professor of Zoology, CVC Intended for students and other novices.
Workshop 4. 3:00 - 4:30 "Digital Imagery: A How-To Approach for Importing, Exporting, Managing, and Everything in Between". Led by Rick McGill, Eastman Chemical Company A. How to design your own image management database system using Microsoft Access. B. How to make your importing/exporting life easier, using some inexpensive commercial software packages.
SOCIAL HOUR: 6:00 - 7:00 (Open bar, Alumni Hall, CVC Campus)
BANQUET: 7:00 - 9:00, Alumni Hall, CVC campus
GASTRONOMIC DELIGHTS Cream of Artichoke Soup Peppered Duck with Chutney glaze, served over wild rice Tenderloin Medalions of Beef, with Burgundy sauce Orange-Ginger baby Carrots Twice-Baked Potatoes Almond Cream custard, with Raspberries White or Red Wine Coffee/Tea
SPEAKER: Dr. Loren W. Knapp, University of South Carolina "Science Education: Coming of Age in the 21st Century"
Friday, October 24, 1997 Room 220 New Classroom Building, Clinch Valley College
8:00 - 8:30 Registration and Coffee 8:00 - 8:30
8:30 - 8:50 Mr. B.J. Craven, Lorillard Research. "Ultrasonic leak detection for the microscopist"
8:50 - 9:15 Dr. Fred E. Hossler, Dept. of Anatomy, School of Medicine, ETSU "Intrinsic lymph nodes in the wall of the urinary bladder - structure and blood supply"
9:15 - 9:35 Mr. Eric Bond, University of Tennessee "The uses of Electron Microscopy in polymer morphology characterizations"
9:35 - 10:00 Dr. Bob Price, School of Medicine, Univ. of South Carolina "The effect of angiotension II on early embryonic heart development"
10:00 - 10:30 BREAK: VISIT THE EXHIBITS
10:30 - 11:00 AReMS Business Meeting
11:00 - 11:25 Dr. Loren W. Knapp, Dept. of Biology, Univ. of South Carolina "Spiculogenesis in Octocorals--Hard tissues: Hard Science"
11:25 - 11:50 Mr. Dave Calvert, Eastman Chemical Company "Of misconceptions and serendipity: Our microscopy and image analysis successes"
11:50 - 12:15 Mr. Mike Kiser, Clinch Valley College "An SEM analysis of development in the tapeworm Hymenolepis dimunata"
12:15 - LUNCH, CVC cafeteria. ---------------------------------------------------------------------- For additional information, pre-registration, and directions, contact Dr. J. Rex Baird, Dept. of Natural Science, Clinch Valley College, Wise, VA 24293 FAX: 540-328-0247 E-mail: jrb-at-clinch.edu Phone: 540-328-0201 AReMS Web site: http://www.clinch.edu/~jrb/arems.html Costs: Registration - Member: $10.00 Non-member: $15.00 Exhibitor: $75.00 Thursday Banquet: $25.00 Friday Lunch: $ 4.00 Annual Dues - Individual: $ 5.00 Corporate: $15.00
Housing: The following motels have reserved a block of rooms at the listed rates. Please mention AReMS when calling. They are side by side in Norton, approximately four miles from Wise.
Holiday Inn (540-679-7000) $54.50 (dbl) Should call before Oct. 15. Super 8 Motel (540-679-0893) $38.59 (1-4 persons) ==================================================================
The FBI Microanalysis Laboratory is seeking development of an application to archive spectra generated by SEM/EDXA, in order to manage large numbers of spectra used for analysis and reference purposes. The application would function somewhat similarly to databases presently used for other spectroscopies, such as FTIR. Database utilities currently provided by EDXA manufacturers consist simply of 1 - naming a spectrum, and 2 - retrieval of spectra by either recalling a specific spectrum by name, or comparing a spectrum to an entire directory of spectra by either a "matching" or quantitative comparison. In order to provide flexible search management, we would like an application that would additionally provide:
1. Attachment of text to spectra. This text would permit descriptors (key words) for each spectrum, which would be searchable with usual "AND/OR" operators, thereby permitting retrieval of spectral clusters based on similar criteria such as material, batch, use, source, date, etc. This spectral database would then function as a true relational database. Other functions, such as sorting, would also be possible. 2. Automatic display of spectra in "nested" fashion (overlayed, but vertically offset slightly), permitting display and critical comparison of numerous spectra simultaneously. 3. Attachment of images to spectra.
I am seeking advice from those who might have experience and/or interest in archiving spectra for the purpose of writing a RFP. If you have interest in the development of a database such as this, please contact me directly for more details.
Thank you. Dennis.
Dennis C. Ward voice: 202-324-2982 FBI fax: 202-324-4018 Microanalysis Laboratory e-mail: DCWard-at-juno.com
} } I heard about a new x-ray microscopy technique that can image cellular } } processes in real-time. Have you heard about this? } } } } } } Bob Clark
Perhaps you are thinking about biospectroscopy, in which you can follow cell processes by their effects on molecular signatures observed by confocal raman spectroscopy. For example, you can follow the movement of a DNA-binding organic molecule (I forget what it was) in 3D and real time as it crosses the membrane, traverses the cell and enters the nucleus. Needs considerable computer power and the 3D effects are calculated after the experiment.
Rosemary White Department of Ecology and Evolutionary Biology Monash University, Melbourne, Victoria 3168, Australia phone 61-3-9905 5670 fax 61-3-9905 5613 email r.g.white-at-sci.monash.edu.au
kszaruba-at-MMM.COM wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } I read an article on X-ray microscopy in the Jan/Feb. 1996 issue } of Biophotonics International (by Laurin Publishing), pg. 58. } The article highlights work by researchers (Dr. Cathie Magowan) } at the Ernest Orlando Lawrence Berkeley National Laboratory, } studying interactions between malarial parasites and host cells. } They studied living [I think] parasites in red blood cells over } 48 hours, in aqueous medium. The stated resolution of the system } was 60 nm in X-ray mode; the microscope could be switched between } visible light and X-ray modes on the same sample. } } If anyone knows more about this I'd be interested out of } curiosity. Being a biologist I don't really want all the gory } details; I just want to know practicle applications, } availability, cost, etc. of the instrumentation. } } Thanks, } Karen } } bengt-at-mail.coxsys.se wrote: } } } } } } } } I heard about a new x-ray microscopy technique that can image cellular } } } processes in real-time. Have you heard about this? } } } } } } } } } Bob Clark } } } } } } Hi ! } } Interesting idea, but it doesn´st seem very likely that it would be } } possible. According to a simple calculation we made, imaging of a cell } } would kill it within a few milliseconds, using x-rays. If someone has other } } information it would be interesting to share it ! } } } } bengt } } Bengt Stocklassa , Managing Director } } Cox Analytical Systems AB | Phone: +46 31 7725300 } } House of Innovations, CTH | Fax: +46 31 7725600 } } 412 96 - Gothenburg, SWEDEN | E-mail: bengt-at-xco.se } } -- } Karen Zaruba } kszaruba-at-mmm.com } 3M Company, 3M Center Bldg. 270-1S-01 } St. Paul, MN 55144 } "The opinions stated above are my own, not necessarily 3M's"Just a quick tag-along to Karen's notes... Having been involved in both applications support and microscope design, I would like to know all the gorey details.
Barbara Foster President Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108 PH: (413)746-6931 FX: (413)746-9311 email:mme-at-map.com --------------------------------------------------------------------------------------------------------------------------------- ********** Microscopy/Microscopy Education ********** America’s First National Consortium of Microscopy Experts Specializing in Customized, On-site Training in all areas of Microscopy, Sample Prep, and Image Analysis
We have seen your message. We at Electron Microscopy Sciences have both a Methyl Methacrylate alone and a Methyl/Butyl Methacrylate kits available. You may see it at our Website where our complete 400 page catalog is up and working at www.emsdiasum or in our hard copy catalog. Please let me know if I may be of further assistance to you.
Sincerely,
Stacie Kirsch Electron Microscopy Sciences http://www.emsdiasum.com
} .... } } The FBI Microanalysis Laboratory is seeking development of an } application to archive spectra generated by SEM/EDXA, in order to manage } large numbers of spectra used for analysis and reference purposes. The } application would function somewhat similarly to databases presently used } for other spectroscopies, such as FTIR. ...
One problem, with respect to other spectroscopies, is the multitude of different SEM/EDX configurations ... for example, take-off angles and detector windows. Operators' choices for instrumental parameters (... e.g., keV ...) also run a gamut. Towards the end of having a "trustworthy" database, you might want to call all of us in for a conference (... any excuse to party ...).
cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
I read warning that " ImagePC beta 1a" , a port of NIH Image to Windows 95 which requires installation of Microsoft's "DirectX, " driver causes computer's display to stop working correctly. I will appreciate feedback or suggestions from those using Image PC.
as many of you will be aware, it can be very difficult to accurately determine diffraction ring diameters in selected area diffraction patterns recorded from multicrystalline TEM specimens. Especially when rings are very spotty.
What I'd like to do is digitize the SAD pattern, locate the exact centre and intergrate pixel intensities through 180 degrees. This will result in a one dimensional diffraction pattern with pairs of spots either side of the undiffracted spot, which should be much simpler to measure.
I would like to know if anyone out there has a computer program to do this type of SAD pattern manipulation. I propose to use Photoshop and a flat bed scanner on a Macintosh to digitize the SAD patterns which would be saved as TIFF or some other suitable format. So I suppose the ideal solution to my problem would be a Photoshop plugin. I also use NIH Image so a plugin for this program would also be suitable.
I would really appreciate any help with a plugin, stand alone program (Mac or PC) or comments on how I should go about writing my own solution. I look forward to your replies,
Mark Blackford TEM Group Materials Division, ANSTO PMB 1, Menai, N.S.W. Australia 2234 Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
John Luft originally suggested 1-2 epoxy propane or propylene oxide as an intermediate solvent for epoxy infiltration because he reasoned that its chemical structure would allow it to be incorporated into the epoxy resin.
But I question if this could ever happen. Consider the boiling points of the three solvents which are being compared:
ethanol....... 78.3 Deg. C acetone....... 56 deg. C. epoxy propane 34.3 Deg. C.
Most epoxies are cured at 70 Deg or above. At this temperature acetone but especially epoxy propane will very rapidly evaporate from the mixture. But alcohol will not. Years back I mixed epoxy resin with 10 percent of each solvent and put the three samples into the curing oven at 70 deg. C. The acetone an epoxy propane mixtures cured perfectly normally. But the alcohol sample stayed sticky. I think it just doesn't evaporate.
Bottom line: since then my lab has always used acetone as a less hazardous alternative to epoxy propane. They both reduce the viscosity of epoxy resin about the same amount. The historically minded could check my 1968 paper-- Viscosity changes in Araldite during polymerization---- Laboratory Practice 17, pp 707-708.
Mel Dickson Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
} I read warning that " ImagePC beta 1a" , a port of NIH Image to Windows 95 } which requires installation of Microsoft's "DirectX, " driver causes } computer's display to stop working correctly. I will appreciate feedback or } suggestions from those using Image PC.
It's tricky. It took 2 installations but it worked fine on my Hitachi lap-top. OTOH, after a bit of work, I opted for ImageTools and removed ImagePC.
I had contact with Chris Cathcart about a DIC system for a Zeiss microscope , but unfortunately he gave me a wrong e-mail address so I can't send him my reply.Knows anybody his correct e-mail.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Reply to: RE} Reference for use of Epon-spurr recipe
Dear Eric, You may want to look up Giammara,B.1993, Scanning 15:82-87 or "The = Microwave Tool Book", Chapter 10, Login and Dvorak, Beth Israel Hospital = Dept. of Pathology, Boston, MA. In addition, Gary Login has published = extensively regarding microwaves and microwave embedding. Linda Chicoine Center for Cell Imaging Yale University New Haven, CT 203-785-3646 http://info.med.yale.edu/cellimg
--------------------------------------
I am trying to to find a reference for the Epon-Spurr resin recipe that = is used for microwave fixation.
I have listed here that the resin can be poylmerized at 70 degree C for 24 hours but I ned a reference that says I can do that..
Any suggestions or does anyone have the reference
Thanks
Eric Fred Hutchinson Cancer Research Center
------------------ RFC822 Header Follows ------------------ Received: by quickmail.yale.edu with ADMIN;16 Oct 1997 14:48:02 -0400 Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com = (8.6.11/8.6.11) id MAA09624 for dist-Microscopy; Thu, 16 Oct 1997 = 12:10:45 -0500 Received: from bug1.fhcrc.org (bug1.fhcrc.org [140.107.10.110]) by Sparc5.= Microscopy.Com (8.6.11/8.6.11) with ESMTP id MAA09621 for = {Microscopy-at-sparc5.microscopy.com} ; Thu, 16 Oct 1997 12:10:42 -0500 Received: from fred.fhcrc.org (fred [140.107.32.6]) by bug1.fhcrc.org (8.8.7/8.8.7) with SMTP id KAA07698 for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 16 Oct 1997 10:21:11 -0700 = (PDT) Received: from localhost by fred.fhcrc.org (SMI-8.6/SMI-SVR4) id KAA21755; Thu, 16 Oct 1997 10:21:09 -0700
This is a multi-part message in MIME format. --------------DED2B0FC0A29C42C04E245BE Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit
Folks: Over the years the subject of immersion oils has come up several times, with the general agreement that Cargille oils are the best. The question is which one! A,B, NVH, FF, and DF Our application is almost entirely fluorescence microscopy. That would be types A, FF, and DF... but which is best and can that selection be used on an inverted scope?
I am sure that someone has done a comparative analysis, if so could you forward the conclusions Thanks
-- Simon C. Watkins Ph.D. Associate Professor Director CBI University of Pittsburgh Pittsburgh PA 15261 tel:412-648-3051 Fax:412-648-2004 URL:http://sbic6.sbic.pitt.edu
--------------DED2B0FC0A29C42C04E245BE Content-Type: text/x-vcard; charset=us-ascii; name="vcard.vcf" Content-Transfer-Encoding: 7bit Content-Description: Card for Simon C. Watkins Content-Disposition: attachment; filename="vcard.vcf"
begin: vcard fn: Simon C. Watkins n: ;Simon C. Watkins email;internet: swatkins-at-pitt.edu x-mozilla-cpt: ;0 x-mozilla-html: TRUE version: 2.1 end: vcard
This is a multi-part message in MIME format. --------------DED2B0FC0A29C42C04E245BE Content-Type: text/plain; charset=us-ascii Content-Transfer-Encoding: 7bit
Folks: Over the years the subject of immersion oils has come up several times, with the general agreement that Cargille oils are the best. The question is which one! A,B, NVH, FF, and DF Our application is almost entirely fluorescence microscopy. That would be types A, FF, and DF... but which is best and can that selection be used on an inverted scope?
I am sure that someone has done a comparative analysis, if so could you forward the conclusions Thanks
-- Simon C. Watkins Ph.D. Associate Professor Director CBI University of Pittsburgh Pittsburgh PA 15261 tel:412-648-3051 Fax:412-648-2004 URL:http://sbic6.sbic.pitt.edu
--------------DED2B0FC0A29C42C04E245BE Content-Type: text/x-vcard; charset=us-ascii; name="vcard.vcf" Content-Transfer-Encoding: 7bit Content-Description: Card for Simon C. Watkins Content-Disposition: attachment; filename="vcard.vcf"
begin: vcard fn: Simon C. Watkins n: ;Simon C. Watkins email;internet: swatkins-at-pitt.edu x-mozilla-cpt: ;0 x-mozilla-html: TRUE version: 2.1 end: vcard
10.30-11.10 Low voltage cryoSEM for x-ray microanalysis Patrick Echlin, Cambridge
11.10-11.50 Variable pressure SEM: an enhancement of cryomicroscopy. Roger Angold, RHM
11.50-12.20 CryoEM and research in cosmetics Philippe Hall=E9got, L=92Or=E9al Recherche
12.20-1.00 Trades Exhibition 1.00-1.45 LUNCH 1.45-2.00 Annual General Meeting
2.00-2.15 Freeze-drying of thin sections for x-ray microanalysis Alice Warley, UMDS, St. Thomas=92 Hospital, London
2.15-2.30 Freeze-substitution strategies for the retention of mineral for= EDX=20 and correlated immunogold labelling of Lowicryl HM20 sections. Jeremy Skepper, Multi Imaging Centre, Cambridge
2.30-2.45 Advances in cryotechniques for analytical microscopy of plant= tissue John Forsdyke, Oxford Microscopy Consultancy
2.45-3.00 Structure of a chaperonin ATP-ase mutant by cryoTEM and 3-D reconstruction. Jose Jimenej, Birkbeck College
3.00-3.30 TEA
3.30-4.00 Cryogenic light microscopy in the development of long term cryopreservation techniques for fungi. David Smith, International Mycological Institute, Surrey
4.00-4.30 Quantitative x-ray mapping of ion-transporting and=20 metal-sequestering epithelia of invertebrate tissues after hyperbaric freezing Carol Winters, University of Wales, Cardiff
4.30 Chairman=92s concluding remarks.
=A325 Registration includes coffee, tea, lunch and Trade Exhibition/Posters welcome
FURTHER INFORMATION FROM : =09
Secretary, CMG Georgina Godwin =09 International Mycological Institute=09 Bakeham Lane, Surrey web site : http://www.gla.ac.uk/Acad/IBLS/II/cryomg.htm=09 TW20 9TY
Tel: 01784470111 x 556 email: G.Godwin-at-Cabi.Org FAX: 01784 470909 Dr Laurence Tetley IBLS EM Centre Joseph Black Building University of Glasgow Glasgow G12 8QQ
email l.tetley-at-bio.gla.ac.uk =09 tel. 0141 330 4431 fax 0141 330 3516 =09 I & I Divisional web pages http://www.gla ac.uk/Acad/IBLS/II/ EM facility web pages http://www.gla.ac.uk/Acad/IBLS/II/em/mcb-em.htm
WinJade from MDI Inc. (XRD-at-AOL.com) is a program for x-ray diffraction analysis that has a software option for doing just that. I've helped them make a few modifications on how they extract the image from the digitized pattern. They now have 4 different ways of doing it. I have submitted an abstract to the International Conference on Metallurgical Coatings an Thin Films-98 being held in San Diego in April on the use of this program with multiphase samples with partial ring patterns. I also plan to publish the paper. If you correlate XRD data with electron diffraction data, this program is great. You can overlay JCPDS or NIST Crystal Database files directly on the image. You can reduce it to a one dimensional pattern that can be overlayed with the XRD patterns. this program has made phase identification from SAD patterns much simpler for me. They have a demo program. I don't know if it has the electron diffraction module in it. You should direct your questions to Quentin Johnson. -Scott
Scott D. Walck PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of S.D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
as many of you will be aware, it can be very difficult to accurately determine diffraction ring diameters in selected area diffraction patterns recorded from multicrystalline TEM specimens. Especially when rings are very spotty.
What I'd like to do is digitize the SAD pattern, locate the exact centre and intergrate pixel intensities through 180 degrees. This will result in a one dimensional diffraction pattern with pairs of spots either side of the undiffracted spot, which should be much simpler to measure.
I would like to know if anyone out there has a computer program to do this type of SAD pattern manipulation. I propose to use Photoshop and a flat bed scanner on a Macintosh to digitize the SAD patterns which would be saved as TIFF or some other suitable format. So I suppose the ideal solution to my problem would be a Photoshop plugin. I also use NIH Image so a plugin for this program would also be suitable.
I would really appreciate any help with a plugin, stand alone program (Mac or PC) or comments on how I should go about writing my own solution. I look forward to your replies,
Mark Blackford TEM Group Materials Division, ANSTO PMB 1, Menai, N.S.W. Australia 2234 Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
Many thanks to the numerous responders to the propylene oxide-alternative. I hope that I did not break off an avalanche of fear because of the carcinogenicity problem. I am aware, that many labs in this country have been using the compound for many years, including my EM-lab. But without wishing to depriciate the potential danger of p.o., I believe that according to the experimental data in rats and mice (literature up to 1996), the compound seems to be a rather weak carcinogen, acting either after repeated local subcutaneous injections (rare sarcomas in rats) or after life-long inhalation of rather high doses ( very low incidence of nasal hemangiocarcinomas in mice). Thus, since p.o. is classified now as a possible carcinogen in humans, we should of course avoid it completely or reduce it to exceptional cases, but nobody should be anxious about exposition in the past. Norbert
What you're describing is similar to a procedure we use called EDIP - electron diffraction image processing. Basically, what we do is scan a TEM negative, digitize it, identify the center of the pattern, then "draw" a series of concentric rings going out form this central spot, increasing by one pixel at a time. We then take the entire integrated intensity inside each of these rings. By subtracting the integrated intensity successively from the next largest ring, i.e., subtract the integrated intensity of ring "Y" from the integrated intensity in ring "Z", then "X" from "Y". etc., one ends up with a "graph" of the diffraction pattern, containing all of the information. This is an extremely useful technique for identifying small precipitates, different phases, etc., particularly if there is a known element or compound included in the pattern which can be used to calibrate the graph. The full procedure is given below - John Philips wrote it up for internal use, so it may be a bit "site specific", but it should give you the useful software names and procedures. NOTE 1: The most recent version of Image Pro Plus contains a circular line profile feature, that does all this automatically. NOTE 2: John Russ of "The Image Processing Handbook" fame has expended a lot of effort on this very problem but from another angle, cf: "Application of the Hough Transform to Electron Diffraction Patterns", Journal of Computer- Assisted Microscopy, Vol. 1, No. 1, pp. 3-37 (1989). I heard that he is producing some software for Photoshop using Hough transforms due out in a month(?) that performs this procedure. Aanyway, good luck! It's a great technique. Please contact me off the listserver if you need more info.
This note describes a procedure for analyzing spots or ring electron diffraction patterns with an image analysis program and a personal computer.
Software: Image analysis program from Image-Pro plus for the PC from Media Cybernetics (www.mediacy.com). Plotting program from Origin Ver 4.0 from Microcal Software Inc.
Hardware: IBM compatible Pentium, Windows NT or equilavent; HP scanner with negative attachment.
Purpose: The analysis is used to do data extraction from electron diffraction patterns that is equivalent to a line scan plot of intensities vs distance (Rd) from the centre of the central spot on the diffraction pattern to some arbitrary user selected point outside the spots or rings near the edge of the photo.
Procedure: The negative to process is converted to a grey level tiff image with a flatbed scanner. A program written in the Auto_Pro macro language of Image-Pro finds the x,y coordinates of the central spot and calculates successive histograms of concentric circular areas of interest starting at the centre of the central spot and increasing by one pixel radius outward to some user-selected point.
Each of these values which are the sums of all the grey levels within these circular areas of interest are appended to a file for later processing with the Origin plotting program. A background image is created from the original using the background extraction feature of the Image-Pro program and a background' file is created using identical xy coordinates and circular histogram parameters. Processing: Subtracting the preceeding value from each of these histogram sums produces a table where each value is equilavent to adding up all the pixel values that lie on the original concentric circle that bounded the histogram. Plotting these values vs their row number is a line scan of intensities from the centre of the central spot to the selected position.(ie: intensity vs Rd). Similar processing of the background image gives a background line profile. Specific instructions: Scan the EDP negative as grey level with the highest resolution available( currently 200dpi on the HP scanner and fine black and white photo mode.). Save as a tiff file. Load the file into Image Pro Invert the file (only to get numbers for a graph that has the background(black) as zero and spots (white) giveing increasing numbers. Make sure you apply the inversion map to the image. select an AOI that includes the spots and rejects edges and various features that are not part of the pattern. duplicate/crop this area and minimize the original image for clarity of the display. create a background image from the duplicated image using the background extraction feature . run the macro diff_main' and follow the instruction on the screen. This macro will find the centre of the image and allow you to select an outer ring where the analysis will stop. This center and outer ring radius will be exactly the same on the foreground' image and the background' image. This is necessary to simplify later processing of the data. The files generated currently have to be edited because the first reading is extraneous and the current version of Image Pro adds two newline characters between readings when you append data to the file. To remove these use for example, Wordperfect and search for three newlines and replace them with one newline. This is essential for Origin or Excel to import the data as a continuous column of numbers. The file as saved has five items in a row and the last one is of interest to us since it is the sum of the grey values contained in the concentric circular histograms that the macro creates. The first four columns can be deleted if desired. The first readings are not accurate representations of the histogram because the size of the circle is only one pixel and it increments by one pixel radius for successive histograms. The first few readings are significant in that their existence defines the distance from the centre of the central spot to where ever the outer ring was chosen. The point of this exercise is to subtract the background image data from the foreground image data and to end up with a plot of Rd vs intensity and try to coorelate the intensities and peak positions with orientation on an unknown sample.
We do a lot of immunofluorescence and after upgrading our scope and looking for very low signals of insitus we under went a study of optimizing mounting media and immersion oils in order to get better signal to noise ratio. I tested every major brand of immersion oil several times and from old and fresh batches. The Cargilles were always highest in autofluorescence and the lowest was Nikon oil. It has always been a mystery why so many people like Cargille, we have several bottles that we gave up using years ago due to it obscuring our signals.
Bob Morphology Core
On Fri, 17 Oct 1997, Simon C. Watkins wrote:
} Folks: } Over the years the subject of immersion oils has come up several times, } with the general agreement that Cargille oils are the best. The } question is which one! A,B, NVH, FF, and DF } Our application is almost entirely fluorescence microscopy. That would } be types A, FF, and DF... but which is best and can that selection be } used on an inverted scope? } } I am sure that someone has done a comparative analysis, if so could you } forward the conclusions } Thanks } } } -- } Simon C. Watkins Ph.D. } Associate Professor } Director CBI } University of Pittsburgh } Pittsburgh PA 15261 } tel:412-648-3051 } Fax:412-648-2004 } URL:http://sbic6.sbic.pitt.edu } }
I've been contacted by a rep from Ted Pella with a new microwave item that allows complete TEM specimen preparation in just 3 hours (live tissue to grid in scope)! Is anyone using this technology regularly, especially for preparing plant tissues? If so, how is it working out for you?
An article in the 27 Sept. '97 issue of New Scientist, pp. 24-28, describes a x-ray microscope being developed at the Rutherford Appleton Laboratory in Oxfordshire, headed by Jie Zhang. The microscope is under development - uses a very bright X-ray laser pulse with a very narrow spectrum, between 2.2 and 4.4 nanometers. Perhaps that laboratory has a web site with more details. Haven't checked; just came across the article in the library.
On Thu, 16 Oct 1997, Bengt Stocklassa wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } =20 } =20 } } I heard about a new x-ray microscopy technique that can image cellular } } processes in real-time. Have you heard about this? } } } } } } =09=09=09Bob Clark } } } } Hi ! } Interesting idea, but it doesn=B4st seem very likely that it would be } possible. According to a simple calculation we made, imaging of a cell } would kill it within a few milliseconds, using x-rays. If someone has oth= er } information it would be interesting to share it ! } =20 } bengt } Bengt Stocklassa , Managing Director } Cox Analytical Systems AB=09| Phone: +46 31 7725300 } House of Innovations, CTH=09| Fax: +46 31 7725600 } 412 96 - Gothenburg, SWEDEN=09| E-mail: bengt-at-xco.se } =20 } =20
Dear Casey, I saw your message. There are many researches here in the United States that have moved over to Microwave technology. It started in Europe and South Africa and now is slowly moving over. The technology is grand and yes even for plant tissue it allows you to do all of the procedures associated with specimen prep for microscopy in the microwave and all in 3 hours or less. This includes dehydration, fixation, staining, polymerizing and evn immuno and decalcification work. We have done alot of work with the major movers who have published on the subject of microwave technolgy, including Gary Login from Harvard, and Dr. Boon who has published the Microwave Cookbook for Microscopists. It truly is a technology worth looking into. There are quite a few laboratory microwave oven manufacturers here in the states and we happen to be one of them. You may see what we have to offer at www.emsdiasum.com or just request a complete technical brochure from us. Please do not hesitate to contact us for further information or if we can give you any technical assistance. Sincerely,
Stacie Kirsch Electron Microscopy Sciences 215-646-1566
Dear colleagues, 8th of October 1997 I posted the following question to the server:
} } Dear all, I greatly should appreciate informations on experiences with existing/ = or on dealers/companies of LASER PROJECTION SYSTEMS (connectable to PC/Video and TV Cams, Remote system). We know a company (ASK-System) in Europe which deals with LIGHT = PROJECTION SYSTEMS for demonstration of slides, PC-data, etc. in teaching and else applications. Has anybody suggestions, informations (company/-ies, approx. price, combinations with periphery, necessary components) for us ? Such a Laser Projection System should work for projection screen = dimensions of at the maximum approx. 2 x 3 m or little less, if available. Any suggestions and comments are welcome. Best wishes for the day sincerely yours Wolfgang MUSS Dept. Pathology LKA, EM-Lab, Muellner Hauptstrasse 48 A-5020 SALZBURG, Austria/Europe Phone: ++43++662+4482-4720 Ext Fax: ++43++662+4482-882 Ext. e-mail: W.Muss-at-lkasbg.gv.at. { {
Unfortunately up to now no suggestion or any answer was received. Since at the time of first posting our server had a lot of troubles and = problems it may be possible that the message was not transmitted in full = or at least garbled. I post our question once more again, asking anybody (also = selling/dealing companies) for suggestions, solutions. Thanking you in advance, have a nice sunday/weekend, Wolfgang MUSS Department of Pathology, EM-Lab. Muellner Hauptstrasse 48 A-5020 SALZBURG, Austria / Europe phone: ++43++ 662 - 4482 - 4720 Ext. Fax: ++43++ 662 - 4482 - 882 Ext. (c/o W. MUSS) e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to -at- is a small "L")
We are planning to purchase a microscope to be used with Polaroid DMC-2000 camera for image analysis of immunohistochemical data. We have found an information on the web about new Nikon Eclipse E-600 and E-800 series microscopes which seem to suit our purposes. But we cannot locate any Nikon dealers in Russian that can provide us with the information on prices and purchase. We can make more complex steps towards purchase through one of the USA companies but can anyone tell what are at least the price ranges for these series of microscopes to know whether they'll fit into our budget?
Thanks in advance.
Konstantin Anokhin
----------------------- neuro-at-redline.ru
Laboratory of Molecular Neurobiology Institute of Physiology Russian Academy of Medical Sciences
Another correspondent claimed that Cargille' is subject to autofluorescense. True, if you are using the wrong type. I have no idea were Nikon is sourcing immersion oil, but you can be sure that several microscope manufacturers are "branding" Cargille's. Here is a copy from our online re immersion oils:
"The refractive index using incandescent light is: Type A 1.5482; type B 1.5468 and type NVH 1.5439. Request additional information if required:
Type B is the most used of these immersion oils. Types A & B have viscosities of 150 and 1250cSt respectively. For a greater gap between cover glass and objective, (or condenser and slide) type B is more desirable. Type A is less sticky and easier to clean up. For horizontal, inverted and inclined instruments and projection equipment, Type NVH with high viscosity of 21,000cST should be used. These three types are classed as low fluorescence, however, for serious fluorescence work types FF and DF - see below - are required.
Type DF combines very low fluorescence and ideal optical properties. It is recommended where specimen fluorescence is good and highest resolution is required. Type FF does not fluoresce but its optical characteristics are not quite perfect."
For fluorescent work the choice seems to be: DF when specimen have fairly strong fluorescence and ideal optical properties are required. If fluorescence is weak type FF should be used which has zero autofluorescence but is optically no quite perfect. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au
} Date: Friday, 17 October 1997 22:35 } } Folks: } Over the years the subject of immersion oils has come up several times, } with the general agreement that Cargille oils are the best. The } question is which one! A,B, NVH, FF, and DF } Our application is almost entirely fluorescence microscopy. That would } be types A, FF, and DF... but which is best and can that selection be } used on an inverted scope? } } I am sure that someone has done a comparative analysis, if so could you } forward the conclusions } Thanks } } } -- } Simon C. Watkins Ph.D. } Associate Professor } Director CBI } University of Pittsburgh } Pittsburgh PA 15261 } tel:412-648-3051 } Fax:412-648-2004 } URL:http://sbic6.sbic.pitt.edu
My name is Myriam Aguirre and I am student of PHD of physics in Argentina. My e-mail is maguirre-at-citefa.edu.ar . I would like to ask you some questions about transmission microscopy. I am studying the structure of compound Mercury Cadmiun Telluride in composition x=0.2 (Hg0.8Cd0.2Te) by TEM. I have had some problems with Hg evaporation. I could not see the structure of the compound because the high temperature which is produced by the electron beam, evaporates the sample. I have tried with the sample cooled with liquid N but the beam goes on eating the sample. Are there solutions to this problem? Thank you in advance. Myriam Aguirre -------------------------------------------------------------------------
I had the same problem with two other Multiples. Pull the heat element out and surround it with some heat sink compound then reinsert it. That should be enough to raise the temp short of buying a new element. Any hardware or electronics store will carry it.
Fred Hayes The Dow Chemical Company Analytical Sciences, Microscopy 1897 bldg, E78 Midland, MI 48667 517-638-2203
} ---------- } From: wise-at-vaxa.cis.uwosh.edu[SMTP:wise-at-vaxa.cis.uwosh.edu] } Sent: Thursday, October 16, 1997 11:31 AM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: LKB Multiplate } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} as many of you will be aware, it can be very difficult to accurately } determine diffraction ring diameters in selected area diffraction patterns } recorded from multicrystalline TEM specimens. Especially when rings are } very spotty. } } What I'd like to do is digitize the SAD pattern, locate the exact centre } and intergrate pixel intensities through 180 degrees. This will result in a } one dimensional diffraction pattern with pairs of spots either side of the } undiffracted spot, which should be much simpler to measure. } This can readily be done using two of the operations in the SPIDER image-processing program. The operation which determines the center coor- dinates and ring radii works by marking from 3 to 20 points on the ring and least-squares fitting the center & radius to those points (for more than 3 selected). Once the center has been found, another operation can be used to calculate the rotational average--which is equivalent to integrating through 360 deg. If you need 180 deg specifically, SPIDER can take half the image, prepare a mirror (or rotated) image and produce a new image which would give you (2 times) the integrated value. This can be done for each half.
} I would like to know if anyone out there has a computer program to do this } type of SAD pattern manipulation. I propose to use Photoshop and a flat } bed scanner on a Macintosh to digitize the SAD patterns which would be } saved as TIFF or some other suitable format.
Several formats (including TIFF) can be converted to SPIDER format.
} So I suppose the ideal } solution to my problem would be a Photoshop plugin. I also use NIH Image } so a plugin for this program would also be suitable. } } I would really appreciate any help with a plugin, stand alone program (Mac } or PC) or comments on how I should go about writing my own solution. I } look forward to your replies,
I have written a stand-alone version (in addition to that incor- porated in SPIDER), and I'll be happy to send you the FORTRAN code. I think you can, then, rewrite it for Mac or PC. Yours, Bill Tivol
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Post-Doctoral Appointment in Electron Microscopy
A post-doctoral appointment for a person skilled in electron microscopy techniques is open in the Materials and Engineering Sciences Center at Sandia National Laboratories-California. The appointee will apply transmission electron microscopy to the characterization of metallurgical and ceramic materials. The candidate must have received a PhD in Materials Science, Chemistry, or Physics. Experience with diffraction contrast methods, HRTEM, and analytical electron microscopy, including EDS and EELS, is strongly desired. Experience using other microscopy methods, including SEM, is also desirable.
Send resume, with names of references, publication list, statements of research expertise, and copies of transcripts to:
Sandia National Laboratories c/o Anna Isham, MS 9111, HR Dept. -CA0041 P.O. Box 969 Livermore, CA 94551-0969
U.S. Citizenship is normally required. Sandia National Laboratories is an Equal Opportunity Employer/Affirmative Action Employer.
For the fourth time a subscribed to the microscopy discussionlist and three times I unsubscribed , because my main interest is LM , I do not work for my research on marine plankton with EM. Would it not possible to make 2 sublists one for LM and one for EM ? Now members who's main work is with LM are sometimes flooded with EM discussions and vice versa. Please give your opinion.
Dear Myriam, } } I would like to ask you some questions about transmission microscopy. I am } studying the structure of compound Mercury Cadmiun Telluride in composition } x=0.2 (Hg0.8Cd0.2Te) by TEM. I have had some problems with Hg evaporation. } I could not see the structure of the compound because the high temperature } which is produced by the electron beam, evaporates the sample. I have tried } with the sample cooled with liquid N but the beam goes on eating the } sample. } Are there solutions to this problem? Thank you in advance.
I have used high voltage (1.2 MV) and low dose imaging to ameliorate this problem. Since the interaction cross-sections with matter fall as the energy of the electron beam increases, the higher the energy (up to ~1 MV), the lower the amount of heat deposited by the beam. This is counter-intu- itive, but none-the-less true. Also, the lower the dose rate, the smaller the ultimate temperature rise. Our scope is equipped with a very sensitive intensified CCD, which allows rapid scanning of the specimen to locate the area of interest, and rapid focussing at low dose rates. Since this camera sacrifices some reso- lution to achieve high sensitivity, we record images on film, and we use LoDose or MRF32 x-ray film to get about one order of magnitude more sensi- tivity than from 4489 or SO163. I cannot tell you that this will be enough for you to be able to get images from your material, but it could be worth doing--at LN2 temperature, of course. Good luck. Yours, Bill Tivol
Greetings, Pieter Houpt wrote: } Dear microscopist, } } For the fourth time a subscribed to the microscopy discussionlist and } three times I unsubscribed , because my main interest is LM , I do not } work for my research on marine plankton with EM. } Would it not possible to make 2 sublists one for LM and one for EM ? } Now members who's main work is with LM are sometimes flooded with EM } discussions and vice versa. The answer here is for folks to make good use of the subject line. The clearer the subject line, the easier it is for us to pick the items to peruse. None of us has the same set of interests. I might read posts on TEM but not ion probe. Someone else may read anything in the life science but nothing in materials science. And so it goes. I think one list with good subject lines will serve the community better than a lot of separate lists. Just my few sou, Tobias Baskin
For my $0.02....I think many of us are involved in both LM and EM, or are interested in both. As long as people stick to using descriptive subject lines, there isn't really a problem sorting through messages to find the ones of direct interest. But isn't it all interesting, anyway? I read (or try to read) the materials stuff...miles/km over my poor head most of the me, but still interesting. Or am I just a hopeless geek?
So - I'd prefer not to see the list split along those lines. Although I could just subscribe to both, if it comes to that; and just deal with the cross-postings.
PLease excuse typing/grammar oddities - I'm midway through a techniques course and losing my marbles.
Tamara CSHL
On Mon, 20 Oct 1997, P.M. HOUPT wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear microscopist, } } For the fourth time a subscribed to the microscopy discussionlist and } three times I unsubscribed , because my main interest is LM , I do not } work for my research on marine plankton with EM. } Would it not possible to make 2 sublists one for LM and one for EM ? } Now members who's main work is with LM are sometimes flooded with EM } discussions and vice versa. } Please give your opinion. } } Pieter Houpt FRMS } } The Hague The Netherlands. }
There is an immediate opening for an experienced SEM operator in our East Coast independent analytical research laboratory. Duties are highly varied and challenging, mostly materials science; the major responsibility is operation of a JEOL JSM-840 with EDS. Frequent client contact. Non-smokers only; drug screen required. Please send resume with salary history in confidence to ablackwood-at-2spi.com
Andy
Andrew W. Blackwood, Ph.D. Structure Probe, Inc. P.O. Box 656 West Chester, PA 19381-0656 Ph: 1 610 436 5400 FAX: 1 610 436 5755 e-mail: ablackwood-at-2spi.com WWW: http://www.2spi.com
I am looking for brief comments on laser printers for doing images from our SEM and CCD cameras.
We have an Apple 4/600PS printer now. It's OK, but slow. I would like to replace it with something like an Apple 12/640PS or HP LaserJet 5M.
Any advice regarding which would be a good choice for our lab (mostly Mac, but printer would be on ethernet) and how much additional memory would be good to get for doing grayscale images of several MBs?
Thanks.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
I am looking for brief comments on laser printers for doing images from our SEM and CCD cameras.
We have an Apple 4/600PS printer now. It's OK, but slow. I would like to replace it with something like an Apple 12/640PS or HP LaserJet 5M.
Any advice regarding which would be a good choice for our lab (mostly Mac, but printer would be on ethernet) and how much additional memory would be good to get for doing grayscale images of several MBs?
Thanks.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
My primary interest is in electron microprobery, nevertheless I feel that I get something out of the biolological EM and even LM material.
After all, most people do put in a "subject" field, and the "delete" button is, on my computer at least, very close to hand.
Please leave things as they are.
Ritchie
} For the fourth time a subscribed to the microscopy discussionlist and } three times I unsubscribed , because my main interest is LM , I do not } work for my research on marine plankton with EM. } Would it not possible to make 2 sublists one for LM and one for EM ? } Now members who's main work is with LM are sometimes flooded with EM } discussions and vice versa. } Please give your opinion.
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
Dear Electron Beamers, Imagers, Probers and etc.'ers. Another great opportunity to touch base with fellow microscopists etc. in the Ottawa area.
WHEN: Wednesday, October 29, 1997, 3:00 p.m. to 8:30 p.m.
WHERE: Same place: RCMP Sgt.s Mess, HQ BLDG., 1200 Vanier Pkwy (at Hwy #417) Directions for the Sgts. Mess and parking available at entrance gate.
FORMAT: Same as before, very informal; any examples of your work in any format would be appreciated for conversation pieces. (Poster board stands will be available). Micrograph Competition "MOST INTERESTING MICROGRAPH"
NOTE: Sorry but no equipment demos: Computer demos can be accommodated to some extent but access to power is limited.
FOOD (free): Snacks, Coffee, Beverages - Served at 4:00 .p.m. Pizza and Subs - Served at 5:30 p.m. For those in need there will also be a cash bar available between 4:00 p.m. and 8:00 p.m.
COST: Its Free=A6 Thanks to some of our local vendors of equipment and supplies. (Who will also be there as colleagues in our field)
I WILL ATTEND (FAX BACK)
NAME PHONE FAX =09 =09 Dave Ballantyne tel:613- 998-6047 fax: 613-952-7325 e-mail: david.ballantyne-at-rcmp-grc.gc.ca Jeff Fraser tel613-: 993-8570 fax:613- 993-8566 e-mail: jeff.fraser-at-nrc.ca
I also would prefer a LM list. Perhaps it is too much work pro bono for the sysop. If list posters would use the prefixes suggested (LM, PLM, SEM, TEM), then it would be much easier to filter out the postings. To enforce this, the sysop would have to have a way to electronically bounce nonconforming postings. Perhaps Nestor would comment.
Cross fertilization is in my opinion essential to science. We all learn from others and I have benefited from the occasional tidbit of information from other fields. It would be a mistake to seperate "microscopists" just based upon a percieved differentiation in the illumination source, imaging modality, or field (Life Science / Physical Science).
I realize that there are a few people that have particuliar interests and they would prefer a seperate list, but it is not my intent to segregate the community but bring it together. With these comment in mind I will respectfully decline the request to create yet another list.
Modern Email programs such as Eudora, have the ability to filter messages based upon key words. To be honest with the volume of mail I receive on a daily basis it is the only way I can keep up. If everyone uses the subject line as it is intended and the delete key then it is a simple job to skip a whole range of messages.
Just for those that have forgotten here is a exerpt from the instructions on Subject lines....
As a courtesy to the readers of this list please indicate in the subject line of your message a reasonable descriptive title of your comment/inquiry. Also preface your description by the conventional abbreviation of the type of microscopy you are interested such as LM, IRM, XRM, TEM, SEM, AFM, STM, uProbe...... For example, if you are interested in optical microscopy and have a question about staining then a Title/Subject line for your message might be
Subject: LM - Need help on Staining Cells
or if you are interested in TEM analysis of dislocations then
Subject: TEM - Dislocation Loop Analysis and so forth.
Just a note about Apple printers. They have a technology called photograde which widens the gray scale and makes photos look better. For details, see their web page at:
http://imaging.apple.com/printers/pr-tech.html
We have that technology on a LaserWriter Pro 630 and I think it prints at 300 dpi as well as a LaserWriter Pro 810 at 800 dpi (but in less time). I do not know if other manufacturers have something like this.
The printer is not especially fast, and I don't know what you can do to really speed one up, except that you might as well put as much RAM in them as will fit.
Disclaimer: I do not sell Apple printers, I just like the ones we have!
Cheers,
John Vetrano js_vetrano-at-pnl.gov _______________________________________________________________________________
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi:
I am looking for brief comments on laser printers for doing images from our SEM and CCD cameras.
We have an Apple 4/600PS printer now. It's OK, but slow. I would like to replace it with something like an Apple 12/640PS or HP LaserJet 5M.
Any advice regarding which would be a good choice for our lab (mostly Mac, but printer would be on ethernet) and how much additional memory would be good to get for doing grayscale images of several MBs?
Thanks.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
} Cross fertilization is in my opinion essential to science. } We all learn from others and I have benefited from the } occasional tidbit of information from other fields. It } would be a mistake to seperate "microscopists" just } based upon a percieved differentiation in the illumination } source, imaging modality, or field (Life Science / Physical Science). } } I realize that there are a few people that have particuliar } interests and they would prefer a seperate list, but it } is not my intent to segregate the community but bring it } together. With these comment in mind I will respectfully } decline the request to create yet another list.
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
Dear Jon, I have been using an HP Laserjet 4P for about four years, now, to print images from my SEM and Quartz PCI. This printer (600 dpi) runs hard all day long and I usually need a new cartridge every three or four weeks from printing everyone's images in 8 X 10 format (instant blow-up). For reliability the HP's can't be beat. It has never given me the slightest problem. I have an additional 4MB of memory, on top of the 2MB it comes with. If you get a 1200 dpi printer, double that. This does an 8 X 10 print in about 1.5 minutes. It is the size of the printout that has the most effect on the speed. You wrote: } Hi: } } I am looking for brief comments on laser printers for doing images from our } SEM and CCD cameras. } } We have an Apple 4/600PS printer now. It's OK, but slow. I would like to } replace it with something like an Apple 12/640PS or HP LaserJet 5M. } } Any advice regarding which would be a good choice for our lab (mostly Mac, } but printer would be on ethernet) and how much additional memory would be } good to get for doing grayscale images of several MBs? } } Thanks. } } Jonathan Krupp } Microscopy and Imaging Lab } University of California } Santa Cruz, CA 95064 } (408) 459-2477 } FAX (408) 429-0146 } jmkrupp-at-cats.ucsc.edu } Regards, Mary
snip=8A NOTE 1: The most recent version of Image Pro Plus contains a circular line profile feature, that does all this automatically. NOTE 2: John Russ of "The Image Processing Handbook" fame has expended a lot of effort on this very problem but from another angle, cf: "Application of the Hough Transform to Electron Diffraction Patterns", Journal of Computer- Assisted Microscopy, Vol. 1, No. 1, pp. 3-37 (1989). I heard that he is producing some software for Photoshop using Hough transforms due out in a month(?) that performs this procedure. Aanyway, good luck! It's a great technique. Please contact me off the listserver if you need more info.
This note describes a procedure for analyzing spots or ring electron diffraction patterns with an image analysis program and a personal computer.
Software: Image analysis program from Image-Pro plus for the PC from Media Cybernetics (www.mediacy.com). Plotting program from Origin Ver 4.0 from Microcal Software Inc.
Hardware: IBM compatible Pentium, Windows NT or equilavent; HP scanner with negative attachment.
Purpose: The analysis is used to do data extraction from electron diffraction patterns that is equivalent to a line scan plot of intensities vs distance (Rd) from the centre of the central spot on the diffraction pattern to some arbitrary user selected point outside the spots or rings near the edge of the photo.
Procedure: The negative to process is converted to a grey level tiff image with a flatbed scanner. A program written in the Auto_Pro macro language of Image-Pro finds the x,y coordinates of the central spot and calculates successive histograms of concentric circular areas of interest starting at the centre of the central spot and increasing by one pixel radius outward to some user-selected point.
Each of these values which are the sums of all the grey levels within these circular areas of interest are appended to a file for later processing with the Origin plotting program. A background image is created from the original using the background extraction feature of the Image-Pro program and a background' file is created using identical xy coordinates and circular histogram parameters. Processing: Subtracting the preceeding value from each of these histogram sums produces a table where each value is equilavent to adding up all the pixel values that lie on the original concentric circle that bounded the histogram. Plotting these values vs their row number is a line scan of intensities from the centre of the central spot to the selected position.(ie: intensity vs Rd). Similar processing of the background image gives a background line profile. Specific instructions: Scan the EDP negative as grey level with the highest resolution available( currently 200dpi on the HP scanner and fine black and white photo mode.). Save as a tiff file. Load the file into Image Pro Invert the file (only to get numbers for a graph that has the background(black) as zero and spots (white) giveing increasing numbers. Make sure you apply the inversion map to the image. select an AOI that includes the spots and rejects edges and various features that are not part of the pattern. duplicate/crop this area and minimize the original image for clarity of the display. create a background image from the duplicated image using the background extraction feature . run the macro diff_main' and follow the instruction on the screen. This macro will find the centre of the image and allow you to select an outer ring where the analysis will stop. This center and outer ring radius will be exactly the same on the foreground' image and the background' image. This is necessary to simplify later processing of the data. The files generated currently have to be edited because the first reading is extraneous and the current version of Image Pro adds two newline characters between readings when you append data to the file. To remove these use for example, Wordperfect and search for three newlines and replace them with one newline. This is essential for Origin or Excel to import the data as a continuous column of numbers. The file as saved has five items in a row and the last one is of interest to us since it is the sum of the grey values contained in the concentric circular histograms that the macro creates. The first four columns can be deleted if desired. The first readings are not accurate representations of the histogram because the size of the circle is only one pixel and it increments by one pixel radius for successive histograms. The first few readings are significant in that their existence defines the distance from the centre of the central spot to where ever the outer ring was chosen. The point of this exercise is to subtract the background image data from the foreground image data and to end up with a plot of Rd vs intensity and try to coorelate the intensities and peak positions with orientation on an unknown sample.
__________________________________________________________________ Philippe Buffat Ecole Polytechnique Federale de Lausanne (EPFL) Centre Interdepartemental de Microscopie Electronique Address: EPFL-CIME, Batiment MX-C, CH-1015 Lausanne, Switzerland Phone: +41(21)693 29 83 Fax: +41(21)693 44 01 (Central European Time) E-mail: philippe.buffat-at-cime.uhd.epfl.ch, WWW URL http://cimewww.epfl.ch/ ______________________________ Eudora F2.1 ___________________________
Hi, despite or better, just because being a greenhorn in using the = Listserver for informations of any +/- EM-related kind I do not vote = for separating the list into several specialized fields of interests. = Nestor Zaluzec=B4s suggestions in my opinion are a great deal.=20 I am learning, learning and learning and am interested to see also other = problems in neighboured areas of my main topic TEM/diagnosis/pathology = (by the way it is too much related and connected with LM due to the = necessity of correlative microscopy for diagnostic purposes).=20 I fully agree to the opinions of Tamara, Tobias, Ritchie and Nestor and = partially to corwinl-at-pt.cyanamid.com in respect to "subject lines" : if = requests would posted using systematic (?) Prefixes, it would help much. = Is there anybody who might have a suggestion how to adhere to "rules" = which must be designed very simple but obligatory??? Therefore: as Ritchie said: "No No No Please leave things as they are."
Best regards, have a nice day Wolfgang MUSS A-5020 SALZBURG, Austria/Europe
Mark Blackford asked for a soft to perform intensity integration along diffraction rings.
If you have access to a copy of Digital Micrograph from Gatan, you can easily add a plug-in named "rotational projection" (or some equivalent name) available on Gatan home site http://www.gatan.com/. It was initially designed to integrate rings in the Fourier transform of images. Thus if you enter your diffraction pattern, let say in TIFF (ie real numbers), you should first tell to the program to change the data into complex numbers and then run the averanging on the rings. If you know how to use the programming language of Gatan, you can also probably modify the data type required in the plug-in (it will then need less RAM to run). Don't forget that diffraction "rings" may deviate from perfect circles by some percent if you don't check and correct the astigmatism in diffraction mode!
Best regards
Philippe Buffat
__________________________________________________________________ Philippe Buffat Ecole Polytechnique Federale de Lausanne (EPFL) Centre Interdepartemental de Microscopie Electronique Address: EPFL-CIME, Batiment MX-C, CH-1015 Lausanne, Switzerland Phone: +41(21)693 29 83 Fax: +41(21)693 44 01 (Central European Time) E-mail: philippe.buffat-at-cime.uhd.epfl.ch, WWW URL http://cimewww.epfl.ch/ ______________________________ Eudora F2.1 ___________________________
Check the review by Kirz et al., Soft X-ray microscopes and their biological applications,1995, Q. Rev. Biophys., 28:33-130. Also, Chris Jacobsen and members of his group at SUNYSB had excellent presentations on the topic, including new developments, at MSA in Cleveland.
Regards, Michel **************************************************** Michel Deschuyteneer, Ph.D. deschuyt-at-sbbio.be Scientist Electron Microscopy Laboratory
SmithKline Beecham Biologicals Rue de l'Institut, 89 B1330 Rixensart, BELGIUM Tel: +32-2-656 9290 Fax: +32-2-656 8164 **************************************************** Standard disclaimer: the opinions expressed in this communication are my own and do not necessarily reflect those of SmithKline Beecham. ****************************************************
Dear all, Does anybody know where I could obtain a sample of natural yttrialite (a thorium bearing yttrium silicate). I would like some to compare with some yttrium disilicate (the synthetic form of yttrialite) that we have made in our lab.
Thanks
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren, Tel: (44) (0) 121 414 3447 IRC in Materials for FAX: (44) (0) 121 414 3441 High Performance Applications, email: I.MacLaren-at-bham.ac.uk The University of Birmingham, http://web.bham.ac.uk/I.MacLaren/ Birmingham B15 2TT, England. ++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
We have a variety of laser printers but my favorite is a Lexmark Optra L(?). It is on a heterogeneous (Mac/PC/UNIX) network and can handle large images without a problem. The printer easily handled a 24MB tiff image embedded into a frame in a Microsoft Word document although it took seven minutes to print. However, I typically print 2-3 640x480 8-bit images on a singe page, along with text, arrows, graphics, etc. and by the time I walk the 50 steps to the printer, the page is done. I'm a happy customer.
Also, check out the latest PC Magazine (Nov. 4) which is devoted almost entirely to printers.
------------------------------------------------ Opinions or statements expressed herein, rational or otherwise, do not necessarily reflect those of my employer.
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 Phone: (508) 750-1717 E-mail: crossman-at-osi.sylvania.com
Our web sites: www.sylvania.com www.siemens.com --
I, too, am in the market for a new printer and have been extremely impressed with the HP and Epson stylus photo printer. It's so new on the market I had trouble finding someone with on to test. Try going through Epson or HP.
I was asked by a chemist to look at carbon nano tubes, as I'm a biologist I have no idea how to prepare the stuff. May I just put some powder on a grid and look at it or are there some special procedures ? Any commments , ideas and references are welcome.
TIA
Marc
------------------------------ SCHMUTZ Marc IGBMC 1 rue Laurent FRIES BP 163 F 67404 Illkirch Cedex FRANCE
Well... I believe that anyone is free to set up a separate listserv f=F6r Light Microscopy without nessecarily having to split the current microscopy listserv...
People are then free to decide what list/lists to subscribe... :)
} } We have a variety of laser printers but my favorite is a Lexmark Optra } L(?). It is on a heterogeneous (Mac/PC/UNIX) network and can handle } large images without a problem.
We also have the Lexmark (Optra R) and use pc/mac/unix with success. The printer is 1200 dpi but the best thing is the grey level capability. Try experimenting with gloss and semi-gloss papers for different appearance.
Chris
Chris Gilpin Biological Sciences Electron Microscope Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 161 275 5170 fax +44 161 275 5171 http://www.biomed.man.ac.uk/biology/emunit/emhome.html
Due to an information of Dalene Josling (thank you very much!) that my = posting was not read-able because of more characters than 80 /line I = post the message once more to the Server (hopefully I was able to get = rid of the wider format, I counted 72 characters/line now):
Hi, despite or better, just because being a greenhorn in using the = Listserver for informations of any +/- EM-related kind I do not vote for = separating the list into several specialized fields of interests. Nestor = Zaluzec=B4s suggestions in my opinion are a great deal.=20 I am learning, learning and learning and am interested to see also other = problems in adjacent areas of my main topic TEM / diagnosis / pathology = (by the way this is too much related and connected with LM due to the = necessity of correlative microscopy for diagnostic purposes).=20 I fully agree to the opinions of Tamara, Tobias, Ritchie and Nestor and = partially to corwinl-at-pt.cyanamid.com in respect to "subject lines" : if = requests would be posted using systematic (?) prefixes, it would help = much (this also would be valid for the "Archives" -section). Is there = anybody who has a suggestion which, and how to adhere to "rules" which = must be designed very simple but obligatory???
Therefore: as Ritchie said: "No No No Please leave things as they are."
Best regards, have a nice day Wolfgang MUSS A-5020 SALZBURG, Austria/Europe
My interests and career have heavily involved both LM and EM, so their separation would mean that I would have to look at two lists instead of one. I hope that they can stay together, since I think they reinforce one another.
A. Kent Christensen Department of Anatomy and Cell Biology Medical Sciences II Building University of Michigan Medical School Ann Arbor, MI 48109-0616 akc-at-umich.edu Tel (313) 763-1287 http://www.umich.edu/~akc/
Dear all, Please point me to some references related to quantitative measurement of lipid/phospholipid/membrane extraction due to conventional chemical processing of biological samples. Thank you in advance.
Yew Meng Heng E.M. Facility Health Sciences Centre McMaster University Hamilton, Ontario Canada
YES there is really such a thing - the Maryland Microscopical Society 25th semi-annual swap meet will be held on November 2, 1997 at the Holliday Inn in Calverton, MD (Exit 29B off Route 95, just North of the DC beltway) 10am - 4pm. 50 dealers expected - this is usually a good show with lots of used equipment. More info.?, contact J.F. Ptak, 202-337-0945.
I have been going for about 10 years, and this is a great place to pick up used equipment.
Instead of using wax for attaching the boats, use nail polish. You can get really cheap nail polish in black, blue, or green if you wish. I gave up on the LKB Multiplates years ago. The temperature is not right for properly drying the 1-micron sections, or staining with toluidene blue either. Joyce Craig Chicago State University
} I was asked by a chemist to look at carbon nano tubes, as I'm a biologist I } have no idea how to prepare the stuff. May I just put some powder on a grid } and look at it or are there some special procedures ? Any commments , ideas } and references are welcome.
We examine carbon nanotubes as follows:
1. suspend the specimen in a small volume of either acetone or methanol (trial and error will be needed to determine the exact dilution. so try different dilutions)
2. suspend the sample by swirling (some people sonicate for several minutes) and use a pipette to transfer a droplet onto a holey, filmed (Butvar or Formvar) and carbon coated grid (suggest purchasing from commercial source)
3. allow to air dry and examine in TEM looking for tubes suspended over the holes (best resolution)
The TEM should be set up for hi res, cooled traps over specimen area and a clean vacuum system is needed.
Good luck.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing 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 that the Microscopy list should not be split. [Nestor needs *something* to keep him busy. :-)! ]
But there already is a list dealing mostly with light microscopy. It's the Histonet list. They do cover related topics as well, but most posts are about light microscopy, immunostaining, and tissues (no materials stuff). It's run by histotechnologists. Address:
{HistoNet-at-Pathology.swmed.edu} Subject: re: subscribe } } WHO SHOULD SUBSCRIBE? } Anyone interested in research or clinical applications of histology, } immunohistochemistry, in-situ hybridization pathology, and electron } microscopy may find Histonet informative and useful. Currently, there are } more than 850 subscribers from all over the world. Subscribers include } hospital employees from major urban centers and obscure remote locales, } university researchers, botanists and the employees of commercial } laboratories, government agencies, veterinary facilities and a wide } variety of commercial industrial ventures. } } WHO RUNS HISTONET? } The list is run by Linda R. Margraf, M.D. and Herb K. Hagler, Ph.D. using } hardware and software owned by the University of Texas Southwestern } Medical School, Department of Pathology in Dallas, Texas. If you have any } questions or problems with Histonet please contact Linda Margraf at } LMargraf-at-childmed.dallas.tx.us. } } HOW DOES THE LIST WORK? } This server, unlike many systems, uses ONLY ONE ADDRESS to send commands } to the computer and to post messages. The server will recognize commands } sent in the SUBJECT line of the message and only when they are spelled } exactly as listed below. Anything not identified as a command will be } circulated to EVERYONE on the list. } } The following is a list of commands the server recognizes: } } subscribe } Your address will be added to the list of subscribers. You will then be } able to send messages to this list that will be forwarded to all other } list subscribers. You will begin to receive all messages sent to the list } by other subscribers. } } subscribe digest } Your address will be added to the list of subscribers who receive a digest } instead of each forwarded message. A digest is a compilation of all the } messages received in a 24 hour period. It is sent to the digest } subscribers every night after midnight. Digest subscribers can post and } respond to messages the same as "real-time" subscribers.
} Well... I believe that anyone is free to set up a separate listserv f=F6= r } Light Microscopy without nessecarily having to split the current microscopy } listserv... } } People are then free to decide what list/lists to subscribe... :) } } / Martin
Hi, I had the same problem, so I can recommend you to make a suspention of this powder in alcohol, then put a drop on a grid with deposited thin film. Contrast will be poor, so you will need to use defocused image.
Vladimir Dusevich dusevich-at-ncsu.edu
Schmutz Marc wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hi around the microscopy world, } } I was asked by a chemist to look at carbon nano tubes, as I'm a biologist I } have no idea how to prepare the stuff. May I just put some powder on a grid } and look at it or are there some special procedures ? Any commments , ideas } and references are welcome. } } TIA } } Marc } } ------------------------------ } SCHMUTZ Marc } IGBMC } 1 rue Laurent FRIES } BP 163 } F 67404 Illkirch Cedex } FRANCE } } Tel: +33 (0)388 653 330 direct } Fax: +33 (0)388 653 201 } email:schmutzm-at-lear.u-strasbg.fr } } ------------------------------
At 03:15 PM 10/21/97 +0900, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I used to view these regularly for a materials scientist. We simply put a small amount of the carbon sample in a solvent and used a sonicator to disperse it. Ten minutes should do a decent job. We then put drops of the suspension onto holey grids which were sitting on filter paper. When they were dry we put them in the TEM for viewing. Some of the nanotubes would sit on the edges of the holes in the film and could be viewed that way.
The nanotubes we viewed required very high magnification and very stable beam and vacuum conditions. Use liquid nitrogen on the pumps and on your decontaminator. Set up your instrument for high resolution conditions and let the electronics and gun stabilize for a while before viewing. Getting the best resolution was trickier by far than preparing the samples.
Good luck.
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
Upon the advice of John MacKenzie, I began training our facility users to open their images in Photoshop, set the halftoning screening to 150 lpi, and using our HP LaserJet 4M+ to make working prints of scanning electron micrographs and, with less success, transmission electron micrographs. They are a vast improvement over the default settings.
However, four weeks ago I bought an Epson Stylus 800 ink jet color printer for home (list $449, cheaper at various outlets). It makes excellent working prints of SEMs and, when pushed to the limit (higest res, expensive paper), makes *nearly* publication quality prints. Fine, light, horizontal lines do show in areas of solid color, although they are less noticeable on a colleague's printer than on my own. They would be great for reviewers' copies and general-purpose applications. The colors (including black and greys) are snappier than on a dye-sublimation printer. (The dye-sub printer I have used shows the horizontal lines, as well, but they are more obscured by the slight blurriness of the process.)
Special ink-jet papers run from a few cents a sheet to about $.50 for the really good stuff. Replacement color and black ink cartridges are about $20 to $27, depending on source, and can be used up pretty quickly on images.
We will be buying this printer for the EM lab within the next week, primarily for good working prints and posters (it does banners, as well). It's a great buy for less than the cost of a dye-sub printer, but I don't expect it to do it all! I still use the darkroom...
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
I don't know if you have a special place in your heart for LaserJet printers specifically, but if you don't, we are getting excellent results with the Epson Stylus Color 800 Ink Jet printer which prints up to 1440 dpi/8 bit image depth. I typically print four 512x512, 8 bit images on a page at 720dpi and it takes about 2 minutes. We are using it on a Novell network with no extra memory. There is special paper for printing -at- 720dpi or above which costs about $12 US for 50 sheets and there is also a special glossy paper which really makes the images look photographic, which sells for about $25 US for 15 sheets. If you have your heart set on a LaserJet printer to use plain paper, I also have used the HP 5P (now no longer sold as the 5P but the 6P has the same engine) which gives true 8 bit gray level shading and excellent pictures. I, personally could not see enlarging the images larger than 2 per page (3.5"x3.5"), but both of these printers have done an excellent job of eliminating the "P" word from the lab. The Epson, being color, has the added advantage of doing a great job with color presentation graphics and transparencies. The Epson sells for about $400 US.
David Bell Millipore Corporation 80 Ashby Road Bedford, MA 01730 (617)533-2108
As usual, I have no vested interest in either of the above mentioned companies, nor do I have anything against Polaroid! (Film may never die!)
Jonathan Krupp wrote:
jmkrupp-at-cats.ucsc.edu on 10/20/97 02:41:30 PM
To: Microscopy-at-Sparc5.Microscopy.Com cc: (bcc: David Bell)
Hi:
I am looking for brief comments on laser printers for doing images from our SEM and CCD cameras.
We have an Apple 4/600PS printer now. It's OK, but slow. I would like to replace it with something like an Apple 12/640PS or HP LaserJet 5M.
Any advice regarding which would be a good choice for our lab (mostly Mac, but printer would be on ethernet) and how much additional memory would be good to get for doing grayscale images of several MBs?
Thanks.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
Ritchie Sims wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } No No No } } My primary interest is in electron microprobery, nevertheless I feel } that I get something out of the biolological EM and even LM material. } } After all, most people do put in a "subject" field, and the "delete" } button is, on my computer at least, very close to hand. } } Please leave things as they are. } } Ritchie } } } For the fourth time a subscribed to the microscopy discussionlist and } } three times I unsubscribed , because my main interest is LM , I do not } } work for my research on marine plankton with EM. } } Would it not possible to make 2 sublists one for LM and one for EM ? } } Now members who's main work is with LM are sometimes flooded with EM } } discussions and vice versa. } } Please give your opinion. } } Ritchie Sims phone: 64 9 3737599 ext 7713 } Department of Geology fax: 64 9 3737435 } University of Auckland } Private Bag 92019 } Auckland } New Zealand
I agree with the concensus about keeping LM and EM together. We have been fighting a battle for more balanced approaches to integrated microscopy and having the two intertwined makes a lot of sense. My original training was with the Royal Microscopical Society, which integrates all types of microscopy. It was the general feeling then, and my continued feeling, now, that each of us has a lot to learn from rubbing elbows with neighboring technologies.
Barbara Foster President Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108 PH: (413)746-6931 FX: (413)746-9311 email:mme-at-map.com --------------------------------------------------------------------------------------------------------------------------------- ********** Microscopy/Microscopy Education ********** America’s First National Consortium of Microscopy Experts Specializing in Customized, On-site Training in all areas of Microscopy, Sample Prep, and Image Analysis
We just make a dispersion of the powder ( say in methanol) and put a drop on a holey grid and we let it dry for a few minutes. We examine the nano tubes at about 100K to 300K. If you use regular carbon coated grids ( rather than holey grids) the image of the nano tubes will not be as clear because the carbon background will interfere.
I was asked by a chemist to look at carbon nano tubes, as I'm a biologist I have no idea how to prepare the stuff. May I just put some powder on a grid and look at it or are there some special procedures ? Any commments , ideas and references are welcome.
TIA
Marc
------------------------------ SCHMUTZ Marc IGBMC 1 rue Laurent FRIES BP 163 F 67404 Illkirch Cedex FRANCE
Question for anyone using a Lexmark Optra printer (specifically Optra Rn+) have you run into "spotting" problems with your prints? We've been having problems with ver fine "spotting" or "speckling" - randomly distributed black dots ranging from ~20 um to 300um (no I didn't actually measure them) all over the page. If so have you managed the a cure? I've clean everything I can think of serval times, without success. I've contacted Lexmark, but they weren't all that helpful and suggested sending it in for servicing. I haven't even gone through one cartridge yet!
Any suggetsions?
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"640K ought to be enough for anybody." -- Bill Gates, 1981
For those of you who print on CMYK printers like the Epson photo, how do you deal with the out-of-gamut colors like pure red or green or blue? It looks like my HP1600 (or is it the program I'm using, Photoshop) may be substituting a single shade of red for all out-of-gamut intensities of red in the original, which makes the print look saturated, lifeless, less detail than I expect. Since I am printing confocal images that are all red & green (& look beautiful on the screen), it's an obvious problem.
Is there a good solution (other than buying an RGB printer)? Is there an easy _automatic_ way to compress the dynamic range to span the colors available from CMYK printers, rather than clipping off the top intensities? or another approach?
Thanks! Richard Richard_Thrift-at-DepoTech.com p.s. it has struck me again how different the images look depending on the monitor used. I optimized the color levels using one monitor and they now look poor on another. Can you recommend color management systems, & indicate price? Thanks!
I've heard a few mentions of the BioQuant system in the past. Now I am considering it myself, and would like to hear a little more detail from users.
What do you like most/least about the system? How easy is it to handle intensity comparisons of stained specimens (BF and Fluor)? What camera option did you choose and how well does it work with a variety of specimen types (macro copy stand, micro BF, fluor, moving subject, etc)?? Does it really do excellent gel densitometry? How about colony counting? 3D measurements? Can it turn VCR tapes (short) into digital movies?
I'd appreciate any comments! Thanks as always, Karen
P.S. Regarding the Message Subject requirements, what about general subjects like digital imaging, printers, etc?? LM vs. TEM, etc. doesn't really fit.
-- Karen Zaruba kszaruba-at-mmm.com 3M Company, 3M Center Bldg. 270-1S-01 St. Paul, MN 55144 "The opinions stated above are my own, not necessarily 3M's"
} We also have the Lexmark (Optra R) and use pc/mac/unix with success. } The printer is 1200 dpi but the best thing is the grey level } capability. Try experimenting with gloss and semi-gloss papers for } different appearance.
In a message dated 97-10-21 18:20:52 EDT, tina-at-pbrc.hawaii.edu, wrote:
{ { However, four weeks ago I bought an Epson Stylus 800 ink jet color printer for home (list $449, cheaper at various outlets). It makes excellent working prints of SEMs and, when pushed to the limit (higest res, expensive paper), makes *nearly* publication quality prints. Fine, light, horizontal lines do show in areas of solid color, although they are less noticeable on a colleague's printer than on my own. They would be great for reviewers' copies and general-purpose applications. The colors (including black and greys) are snappier than on a dye-sublimation printer. (The dye-sub printer I have used shows the horizontal lines, as well, but they are more obscured by the slight blurriness of the process.)
Special ink-jet papers run from a few cents a sheet to about $.50 for the really good stuff. Replacement color and black ink cartridges are about $20 to $27, depending on source, and can be used up pretty quickly on images. } }
Have you tried Kodak Inkjet Photographic Quality paper yet? It works out at about 60 cents a sheet and indeed produces "near photographic quality" results.
I have tried everything available and am very pleased with this product.
I use it with a Hewlett Packard 694C DeskJet printer but I believe that it is compatible with the Epson Stylus printers also.
} Due to an information of Dalene Josling (thank you very much!) } that my posting was not read-able because of more characters than 80 /line } I post the message once more to the Server (hopefully I was }
For those who use Pegasus mail, there is a option "Reader as you are reading a message with thw option "Wrap long lines" very useful.
} "No No No } Please leave things as they are."
Since I am working in multidissiplinary environment which includes SEM, TEM, CONF, LM, EDS and hopefully STEM, and PEELS at a later stage. We do have users from Life sciences and Physical sciences and a separated list is unthinkable for me.
My 5 cents worth (SA) ~ 1.3 cents worth (US). Bad exchange rate!
A couple of people e-mailed me to thank me for mentioning the trick of increasing the halftone screening resolution, so I will repeat it for you all.
In Photoshop, under File select Page Setup..., then click on Screen... *Uncheck* Use Printer's Default Screen, and enter 150 lines/inch for Frequency. Leave Angle at 45 degrees and Shape as diamond, unless you want to experiment.
To set this as the new default for Photoshop, Alt-click on the Save button (Windows) or Option-click (I think) on Save for Mac.
I have found this trick to work on numerous printers, not just HP LaserJets. It even made my Brother fax/copier/scanner/200dpi printer at home print reasonable SEMs! For some of the printers it was more necessary to adjust gamma levels or just to lighten up the images than on others, as more ink will be applied.
This tip originally came from John MacKenzie at MSA in Cleveland, and was worth the entire cost of traveling all the way from Hawaii!
I've got a few fun tricks coming up in the next Microscopy Today, and a more complete Photoshop tutorial in another month or so. I'd be glad to incorporate any other tips you all may have, so don't hesitate to e-mail.
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangela **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Regarding the suggestion that the List Server should be be sub-divided into LM andEM. No! No ! No! Microscopy is a broad canvas and we can learn from all aspects of the subject and fromm different application disciplines within microscopy.
Patrick Echlin Multi-Imaging Centre Cambridge University
On Mon, 20 Oct 1997, P.M. HOUPT wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear microscopist, } } For the fourth time a subscribed to the microscopy discussionlist and } three times I unsubscribed , because my main interest is LM , I do not } work for my research on marine plankton with EM. } Would it not possible to make 2 sublists one for LM and one for EM ? } Now members who's main work is with LM are sometimes flooded with EM } discussions and vice versa. } Please give your opinion. } } Pieter Houpt FRMS } } The Hague The Netherlands. }
Marc, The method I used was to disperse the powder in 2-propanol. Sonication helps but is not necessary. Then a drop of the suspension is dried on a holey carbon support film. If you do not use a holey carbon, you won't be able to distinguish the tubules from the background.
Regards, Eric Lachowski
---------------------- Dr Eric E. Lachowski University of Aberdeen Department of Chemistry Meston Walk Old Aberdeen AB24 3UE Scotland +44 1224 272934 e.lachowski-at-abdn.ac.uk
Does anybody have a spare Pirani gauge for an Edwards high vacuum carbon coating unit E12E. I would be eternally grateful to any list server member who did or new how I could get hold of one (relatively cheap). Thanks
The Microscopical Society of Southern California (MSSC) is proud to invite anyone interested to a lecture by Brian J Ford, on October 28 (Tuesday)
Topic: Antony van Leeuwenhoek and the Single Lens Microscope
Our program this month features a presentation on Antony van Leeuwenhoek (1632-1723) and single lens microscopy. Professor Ford is the author of countless books and papers and has done extensive research on the historical development of microscopy, including work with original, 17th century Leeuwenhoek instruments and samples from the archives of the Royal Microscopal Society in London. Many American microscopists know Professor Ford through his annual presentations at Inter/Micro in Chicago. The (MSSC) is Located at the Crossroads Schools in Santa Monica. Crossroads is located on Olympic Boulevard and 20th street. Anyone interested may contact Larry Albright the Program Chair at: 310-399-0865 Days or 310 471-0424 evenings.. It will be an exiting evening.......Larry
"The beauty and genius of a work of art may be reconceived, though its first material expression be destroyed, but when the last individual of a race of living things breathes no more another heaven and another earth must pass before such a one can be again." William Beebe albrite-at-Plasma-Art.com 419 Sunset Avenue Venice CA 90291 310-399-0865 310-392-9222 FAX
You are invited to check out my web site at: www.Plasma-Art.com
} The Microscopical Society of Southern California (MSSC) is proud to invite anyone } interested to a lecture by Brian J Ford, on October 28 (Tuesday) } } Topic: Antony van Leeuwenhoek and the Single Lens Microscope } } Our program this month features a presentation on Antony van Leeuwenhoek } (1632-1723) and single lens microscopy. Professor Ford is the author of } countless books and papers and has done extensive research on the } historical development of microscopy, including work with original, 17th } century Leeuwenhoek instruments and samples from the archives of the } Royal Microscopal Society in London. Many American microscopists know } Professor Ford through his annual presentations at Inter/Micro in } Chicago. } The (MSSC) is Located at the Crossroads Schools in Santa Monica. } Crossroads is located on Olympic Boulevard and 20th street. } Anyone interested may contact Larry Albright the Program Chair at: } 310-399-0865 Days } or 310 471-0424 evenings.. } It will be an exiting evening.......Larry } } "The beauty and genius of a work of art } may be reconceived, though its first material expression be destroyed, but when the last individual of a race of living things breathes no more another heaven and another earth must pass before such a one can be again." William Beebe } albrite-at-Plasma-Art.com } 419 Sunset Avenue } Venice CA 90291 } 310-399-0865 } 310-392-9222 FAX } } } You are invited to check out my web site at: www.Plasma-Art.com }
Jonathan Swift 1733
So, naturalists observe, a flea Larry Albright Hath smaller fleas that on him prey; albrite-at-Plasma-Art.com And these have smaller fleas to bite'em, 419 Sunset Avenue And so proceed ad infinitum. Venice CA 90291 Thus every poet, in his kind, 310-399-0865 Is bit by him that comes behind. 310-392-9222 FAX Web Page.www.Plasma-Art.com Jonathan Swift 1733
I used to use a pyramitome to trim my blocks for TEM, and since I have relocated to another facility, I don't have the luxury of using it anymore. I was wondering if they are still available and would appreciate more information on getting one. Please reply to me at my email address, rather than cluttering up the listserver.
Thanks in advance,
Susan
Susan Carbyn Atlantic Food and Horticulture Research Centre Agriculture and Agri-Food Canada Kentville, Nova Scotia B4N 1J5 Canada
Recently our resident molecular biologists have requested us to fix and process suspensions of ovarian granulosa cells so that they can perform in situ hybridisations on them. Because of the limitations of the experiment and the in situ methodology this locks us into a number of factors. 1. the cells must be in suspension (ie they have been mechanically dispersed) 2. Paraformaldehyde must be the fixative 3. the tissue must be embedded in paraffin
We have tried fixing in suspension in 4% paraformaldehyde in phosphate buffered saline, centrifuging to a pellet, resuspending in a small volume of 2% agar followed by 8 hours of dehydration (ethanol), clearing (xylene) and infiltration/embedding in paraffin. However there appears to be a lot of damage to the cytoplasm and the nucleii are very condensed.Any thoughts at all on preparing cell suspensions would be appreciated.
Regards Peter Smith AgResearch Wallaceville Upper Hutt New Zealand Smithp-at-agresearch.cri.nz
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I use all forms of microscopy and would be more than a bit annoyed if I had to subscribe to multple lists. In spite of its being somewhat cumbersome I prefer the listserver to remain unchanged.
my $0.02(US) worth
Edward J. Basgall, PhD The Pennsylvania State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618 http://www.personal.psu.edu/ejb11/
Privilege does not absolve one of ecological responsibility.
Greetings, Peter Smith asked alternatives to agar embedding for "loose" samples. We have had good luck with a trick we picked up from the cryofixation folks. We make wire loops out of very fine copper wire (36 gauge) and coat the loop with Formvar. The diameter of the loop can be a few mm or up to 7mm (we have never tried larger). You then collect your cells on the Formvar. For example, if the cells are in a small volume of solution, go "fishing" with the loop. You can make the Formvar surface more sticky by pretreatment with polylysine. Now what you do is to place a second Formvar coat over the loop, thus trapping your sample between two Formvar films. You will probably need to be sure that you don't have too great a puddle of liquid on your loop for this step. Some fiddling will be needed. We cast small rectangles of Formvar on water, with the narrow side of the rectangle a bit wider than the loop diameter, and the long side of the rectangle a bit longer than twice the loop diameter. Then you can very quicky dunk your loop with the samples onto the Formvar rectangle. Line up your loop with the middle of the rectangle, so it makes two squares, and plunge at right angles to the water. The Formvar rectangle just snaps to the loop. Now your cells are trapped inside. But the FOrmvar is readily permeable to your fixative, to your dehydration agent and even to paraffin. At the end of the "day", you can excise the loop the with a razor. In fact, if you really get 36 gauge copper, you can just cut through it with a double-edged razor blade. I hope this helps. If any of the above was foggy, please reply. Tobias Baskin
} Recently our resident molecular biologists have requested us to fix and } process suspensions of ovarian granulosa cells so that they can perform } in situ hybridisations on them. Because of the limitations of the } experiment and the in situ methodology this locks us into a number of } factors. } 1. the cells must be in suspension (ie they have been mechanically } dispersed) } 2. Paraformaldehyde must be the fixative } 3. the tissue must be embedded in paraffin } } We have tried fixing in suspension in 4% paraformaldehyde in phosphate } buffered saline, centrifuging to a pellet, resuspending in a small } volume of 2% agar followed by 8 hours of dehydration (ethanol), } clearing (xylene) and infiltration/embedding in paraffin. However there } appears to be a lot of damage to the cytoplasm and the nucleii are very } condensed.Any thoughts at all on preparing cell suspensions would be } appreciated. } } Regards Peter Smith } AgResearch Wallaceville } Upper Hutt } New Zealand } Smithp-at-agresearch.cri.nz }
To those more chemically minded than myself (i.e. everyone),
I've seen many notices that waste 2% osmium tetroxide can be reduced to less hazardous form by mixing with twice volume of corn oil. I'd like to implement this practice, but I also use a lot of 0.5% ruthenium tetroxide. Can this also be reduced with oil? The only thing I've heard of for RuO4 is sodium bisulfite, which is quite toxic/harmful itself. It would be great to find a single practice that works for both Os and Ru.
Also, does anyone know if buffer solutions (cacodylate or phosphate) interfere with OsO4 reduction by corn oil?
Thanks for your help, Karen
-- Karen Zaruba kszaruba-at-mmm.com 3M Company, 3M Center Bldg. 270-1S-01 St. Paul, MN 55144 "The opinions stated above are my own, not necessarily 3M's"
may-be this question doesn't belong here, but I am totally stuck so I still ask it: I have recently taken some 120 Mb of digitized pictures with a CCD-camera equiped microscope using IPlab software on a MacIntosh. Does anybody know a windows program that can read IPlab's TIFF format (12 bit greyscale, stored in 2 bytes, I think) and convert it to any windows-recognized format? These data are important for me. Thanks in advance. Kees Jalink
--
Kees Jalink The Netherlands Cancer Institute, dept. of Cell Biology H1 Plesmanlaan 121 1066CX Amsterdam, the Netherlands 020-5121982 (tel) / 020-5121989 (fax) kees-at-NKI.NL (email) / 0297-320248 (tel at home)
Dear Karen, for over ten years now I (use my OsO4-solutions) and dispose of my used = up working solutions in a way which is very safe, but uncommon. I=B4ve = had posters on that at MSA-Meeting Boston 1992 (also included in the = Proceedings, 50th Ann. Meeting, p. 754/755: MUSS W.H.: "Pro=B4s for a = multiple/repeated use and safe disposal with recovery of OsO4 = solution(s) in routine EM") For about 1300-1600 specimen blocks to osmicate I need only 4-6 g = OsO4/year. I don=B4t use the cornoil-disposal method, because I think Os a valuable = metal which shouldn=B4t be wasted off the drain/ down the sink ("with a = plenty flush of water" !! as has been proposed by several people, which = in my opinion would not fit ecological considerations!) but be recovered = and recycled. I learned my lession at the 1992 Boston Meeting as well as within the = last 5 years: I tried people to convince that it is possible to reduce = the amount of OsO4 used for staining/postfixation of specimens AND = recover all of the Os from solutions as used (like buffers, like = mixtures etc.) as this is possible in Switzerland, where a regular Os = recovery and recycling has been established for all EM-Labs aided by the = SGOEEM=3DSwiss Society for EM since 1972!!=20 It is not easy to convince people about that though the process is very = easy, simple, effective in my experience and more or less costs are low. = I am doing it. I have, unfortunately, no experience in "deactivating" = RuO4, but I think, considering my scanty knowledge from chemistry = lessions and 17 years experience in disposal/recovery of OsO2/Os (+/- = pure), this should be no problem at all. If you want to get informed about the reasons, why and how I do it: = please don=B4t hesitate to contact me by e-mail.
Best wishes for a lucky day to you and all of the community
Wolfgang MUSS EM-Lab, Dept. Pathology, LKA A-5020 SALZBURG, Austria/Europe.
I fix cells in suspension on regular basis and use agar to encapsulate the cells before pelleting them. Fix cells in suspension, spin down gently to remove excess fix and resuspend in minumum fix. Make 2% agar and keep at 48 deg to avoid solidifying the agar. Mix equal vol of concentrated cell suspension and agar, solidify the suspension and then run up the agar piece as you normally would any tissue.... hope this helps. However if the tissue is losing some of the morphological integrity then you may need to change the fixative or may be add low concentrations of glut to it. Neelima Shah,UOP morphology core.
At 08:47 AM 10/23/97 +1300, Smith, Peter wrote: } } Recently our resident molecular biologists have requested us to fix and } process suspensions of ovarian granulosa cells so that they can perform } in situ hybridisations on them. Because of the limitations of the } experiment and the in situ methodology this locks us into a number of } factors. } 1. the cells must be in suspension (ie they have been mechanically } dispersed) } 2. Paraformaldehyde must be the fixative } 3. the tissue must be embedded in paraffin } } We have tried fixing in suspension in 4% paraformaldehyde in phosphate } buffered saline, centrifuging to a pellet, resuspending in=A0 a small } volume of 2% agar followed by=A0 8 hours of dehydration (ethanol), } clearing (xylene) and infiltration/embedding in paraffin. However there } appears to be a lot of damage to the cytoplasm and the nucleii are very } condensed.Any thoughts at all on preparing cell suspensions would be } appreciated. } } Regards =A0=A0=A0=A0 Peter Smith } =A0=A0=A0=A0=A0=A0 AgResearch Wallaceville } =A0=A0=A0=A0=A0=A0 Upper Hutt } =A0=A0=A0=A0=A0=A0=A0 New Zealand } =A0=A0=A0=A0=A0=A0 Smithp-at-agresearch.cri.nz=A0=A0=A0=A0=A0 } =A0=A0=A0=A0=A0 } } Attachment Converted: "c:\eudora\attach\LM cell suspensions"=20
Has anybody evaluated the Agfa T5000 or T8000 or the ScanMate 11000, 3000, F8 or F8 Plus for use in high resolution EM (2D crystals, helical arrangements or singe particles of proteins)? We're trying to decide which scanner to buy and we're specifically looking at geometric accuracy of the scan and modulation transfer functions. So far we have data on the Zeiss SCAI, the Scitex Eversmart Pro and an Eikonix scanner.
-- Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 86 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www_scem.csb.ki.se/pages/philip.html
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have had all of my nerve and muscle questions answered in; Myology by Andrew G. Engel and Betty Banker, Vol I and II, McGraw-Hill. I have no idea of the year of the latest edition or the price. My edition (1986) has 2106 pages. Kate Connolly
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Reply to: RE} molviol distributor?
Dear Reinhard:
We buy our supply of Mowiol from Calbiochem (LaJolla, CA 92039) = Tel.1-800-854-3417 The catalogue number is 475904 and 100g costs $44US. Hope this helps.
Linda Chicoine Center for Cell Imaging Yale University New Haven, CT 203-785-3646 http://info.med.yale.edu/cellimg
--------------------------------------
Hi,
I am looking for the fluorescent antifading mounting medium MOVIOL. Can someone out there tell me who the distributor is?
------------------ RFC822 Header Follows ------------------ Received: by quickmail.yale.edu with ADMIN;23 Oct 1997 05:18:36 -0400 Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com = (8.6.11/8.6.11) id DAA01154 for dist-Microscopy; Thu, 23 Oct 1997 = 03:38:17 -0500 Received: from duchesse.zdv.Uni-Mainz.DE (duchesse.zdv.Uni-Mainz.DE = [134.93.8.122]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id = DAA01151 for {microscopy-at-sparc5.microscopy.com} ; Thu, 23 Oct 1997 = 03:38:15 -0500 Received: from localhost (windoff-at-localhost) by duchesse.zdv.Uni-Mainz.DE (8.8.5/8.8.5) with SMTP id KAA13732; Thu, 23 Oct 1997 10:48:56 +0200 (MET DST) X-Authentication-Warning: duchesse.zdv.Uni-Mainz.DE: windoff owned = process doing -bs
What makes you think that sodium bisulfate is toxic? The Merc Index gives its LD50 (in rats) as 115 mg/Kg, and says that it is used as a preservative and bleach in food.
best regards mark
Mark W. Lund, PhD Director } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"The state is good at simple tasks, like killing people and seizing their wealth. It has far more trouble reaching inside individuals and making them good." Doug Bandow
You might want to begin with Image Alchemy from Handmade Software (http://www.handmadesw.com/index.html) or Thumbs Plus from Cerious Software (http://www.cerious.com). While I don't know if they have direct translators for IPLab files, they can handle many types of conversion.
NIH Image may work also. I don't have the URL handy. Does anybody else have it?
If you can't translate the files directly, can you use IPLab to translate them into an intermediate, platform-independent format? TIFF? JPG? Photoshop?
------------------------------------------------ Opinions or statements expressed herein, rational or otherwise, do not necessarily reflect those of my employer.
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 Phone: (508) 750-1717 E-mail: crossman-at-osi.sylvania.com
Our web sites: www.sylvania.com www.siemens.com --
Thanks a lot to all who gave me some nice tricks to observe carbon nanotubes. It helped me a lot and due to your nice indications I've got correct images at the first shot.
Marc
------------------------------ SCHMUTZ Marc IGBMC 1 rue Laurent FRIES BP 163 F 67404 Illkirch Cedex FRANCE
In a message dated 97-10-23 03:19:53 EDT, Kees Jalink wrote:
{ { Does anybody know a windows program that can read IPlab's TIFF format (12 bit greyscale, stored in 2 bytes, I think) and convert it to any windows-recognized format? } }
PaintShopPro, ver. 4.14, from JASC will read 6 different TIFF subformats. These are: uncompressed, Huffman compresssed, pack bits compressed, LZW compressed, Fax Group 3 compressed, and Fax Group 4 compressed. None of these are 12 bit grayscale. They range thru 1, 4, 8, and 24 bit/pixel color scales. Attach a single file to an e-mail to me and I will try to change the image to something else, such as a gif or a jpg, your choice. PSP supports 35 different formats.
Yours truly, Steve Stokowski Stone Products Consultants Concrete Petrographers 10 Clark Street, Suite A Ashland, Massachusetts, 01721 USA 508-881-6364 http://members.aol.com/CrushStone/index.htm
My safety officer has told me that treating the osmium as noted may bring charges of reprocessing hazardous waste, for which the University doesn't have a license. They insist I give them the Osmium as is for disposal.
} To those more chemically minded than myself (i.e. everyone),
} I've seen many notices that waste 2% osmium tetroxide can be } reduced to less hazardous form by mixing with twice volume of } corn oil. I'd like to implement this practice, but I also use a } lot of 0.5% ruthenium tetroxide. Can this also be reduced with } oil? The only thing I've heard of for RuO4 is sodium bisulfite, } } which is quite toxic/harmful itself. It would be great to find a } single practice that works for both Os and Ru.
} Also, does anyone know if buffer solutions (cacodylate or } phosphate) interfere with OsO4 reduction by corn oil?
} Thanks for your help, vKaren
} -- } Karen Zaruba vkszaruba-at-mmm.com } 3M Company, 3M Center Bldg. 270-1S-01 vSt. Paul, MN 55144 } "The opinions stated above are my own, not necessarily 3M's"
--------------------------------------------------------------------- COME TO OUR OPEN HOUSE "INNER SPACE/OUTER SPACE" IT IS FREE, AND EVERYONE IS WELCOME Saturday, November 15, 1997 4-8 pm SEE ELECTRON MICROSCOPES TELESCOPES AND IN OPERATION FOR MORE INFORMATION, SEE MY WEB PAGE OR CALL 594-6182 --------------------------------------------------------------------- Dr. Steven Barlow, Associate Director EM Facility/Biology Department 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/EM_Facility
} windoff-at-mail.uni-mainz.de Reinhard, MOWIOL Cat.#475904, is distributed in the USA by CALBIOCHEM-NOVABIOCHEM Corp. of LaJolla, CA phone 800 628 8470
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Salut to the listmembers, } } may-be this question doesn't belong here, but I am totally stuck so I } still ask it: } I have recently taken some 120 Mb of digitized pictures with a } CCD-camera equiped microscope using IPlab software on a MacIntosh. Does } anybody know a windows program that can read IPlab's TIFF format (12 bit } greyscale, stored in 2 bytes, I think) and convert it to any } windows-recognized format? These data are important for me. } Thanks in advance. } Kees Jalink } } -- } } } Kees Jalink } The Netherlands Cancer Institute, dept. of Cell Biology H1 } Plesmanlaan 121 1066CX Amsterdam, the Netherlands } 020-5121982 (tel) / 020-5121989 (fax) } kees-at-NKI.NL (email) / 0297-320248 (tel at home) } You might try Wang Image (now a standard free imager that comes with windows 95, or is availiable as freeware I believe) We use it to convert JEOL tiffs to tiff formats that Lview/NIH can read. It seems to read many different tiff formats but only saves in one (which I don't know, but a tiff saved by wang image suddenly becomes readable by all of our other viewer programs). \ *shrug* hope it helps} Christopher Institute of Meteoritics Albuquerque, NM USA
An issue that won't go away? I think there are legitimate reasons for this.
Like the sys-op and others with similar views expressed here, I believe in cross-fertilization among the hosted disciplines, and at least skim each message routed through the MSA Server.
Nevertheless, I do appreciate the other view. All inclusiveness is nice, but we all have different depths of interest, time, and patience.
In addition, MSA Mail Reflector messages arrive jumbled with more pressing and time sensitive professional and personal mail. The messages are rarely labeled in the subject field with the suggested key-words, and none bear a prefix (e.g. "MSA:") identifying them as emanating from this forum. I wade through it all-at-once, unable to apportion time by priority &| interest. How many have discarded a critical bill or letter intermingled with the mountain of trash stuffed in the post box? Need that experience be re-enacted in the electronic medium?
A fix does exists in most e-mail browsers in the form of the recipient defined filters (electronic or visual); some e-mail might get redirected to the MSA\LM folder, some to MSA\EM folder, some to MSA\Bio folder, ..., some to MSA\MISC folder, ... - to be perused as time and interests allow, and the reminder to the TrashBin folder. Without those key words, though, it's tough to set up the filters adequately.
However, the sys-op's recent appeal for voluntary use of the key-words in the subject field (please excuse this, Nestor) is naive; after a brief excursion into compliance, most of us return by expediency to the minimum that will get by. As the MSA Mail Reflector is currently set up, there are no simple solutions to the problem.
Suggested changes to the MSA Mail Reflector to address these issues:
(1) Filter the incoming postings for approved key-words in the subject field (for inclusiveness, "MISC." among them) and bounce back non-compliant mail with appended reminder of the policy and a list of acceptable key-words to choose among. (A new key-word may be added to the approved list when the traffic on the issue warrants separating it from the "MISC".) The filtering should also effectively purge the forum of machine generated SPAM that may spill in here inadvertently.
(2) Automatically prefix the subject field with the "MSA:" key-word. This, above the current practice by the MSA Server of inserting the MSA logo into the body text, will aid the subscribers in determination of the source and in sorting of e-mail prior to wading into the content.
(3) Create and ENFORCE a policy for vendors to use "VENDOR" as perhaps the last key-word in the subject field. Such feature would permit the commercial members to maintain their valuable contributions without bothersome censorship, but with ample warning for the rest of us. I am tiring of tripping over the same outfit indulging here in thinly disguised attempts at brand recognition & promotion; a disclaimer at the end of the message is too little too late. There ought to be a recognized, regulated, and thus less insidious venue for this stuff. (If non-compliance continues unabated, a special filter at the server might be programmed to append the VENDOR key-word automatically, perhaps with addition of the vendor's name, for the recalcitrant few.)
I do appreciate that all of this would fall on Nestor, who can't be looking for more work at the moment. But, in my opinion, it would address the root causes for the recent gripes without inconveniencing those who like things just the way they are.
So much for my 50 cents worth.
Otherwise, a great service, and under no circumstances I would unsubscribe.
A few weeks ago I posted a querry to the listserver wondering why I was not getting good fluorescent labeling using otherwise successful primary antibodies applied to the surface of one micron thick LR White sections followed by secondary TRITC or FITC conjugates. I sincerely thank those that responded, and I am now sharing the responses:
I have tried labelling 1 micron sections with lectins conjugated with TRITC and they worked exceptionally well.... that is the lectin was labelled with the TRITC tag...
Eric Rosen
I suspect the problem lies in the different secondaries, maybe try the fluorescent-gold-secondaries that are commercially available to test fluorescence and gold label at the same time. The product is called fluoronanogold and is available from Nanoprobes Inc. You can tap into their website (http://www.nanoprobes.com) to obtain detailed protocols or contact them at 516-444-8815. They also put out a newsletter.
Margaret Springett e-mail hukee.margaret-at-mayo.edu IEM Specialist at Mayo Foundation 1426 Guggenheim Rochester, Mn. 55905
I routinly test 1 micron LR White sections on glass before going to the trouble of Immuno EM. I usually use peroxidase ABC which seems to enhance the signal enough to be visualized. However, the fluorescent signal is very weak unless you start stacking antibodies which can create more backround. So you need a very good optimized scope to be able to see the signal and your faint signal may be obscured by the backround signal of the aquamount which has higher fluorescence than some others. Prolong from M olecular Probes is very quiet and enhances Texas Red. Or my favorite for quiet and quick is: 70% Glycerol, 25% .5M Tris pH 9, 5% n-Propylgallate, heat to mix then pH to 7.4 and store in the fridge. For use: 1 drop, swirl, drain most of it off then cove rslip. There should be barely enough to cover the coverslip.
Good Luck Doug
Robert Underwood Morphology Core Dermatology U of Wash. Seattle, WA
We had the same disappointment a few years back when we went from great IHC labelling at the EM level to nothing with half micron LR sections. We got round the problem in two ways. One, we gold labelled the half micron LR sections then silver intensified and photographed with white light. Two, we went back to embedding in wax, cut 5 to 10 micron sections, de-waxed and got excellent FITC-IHC labelling. Although we were reticent to go to wax embedding (19th century technology) it rendered available such enormous areas of antigenic sites that the IHC was excellent plus we could use the sections for in situ work and the fluorescent apoptosis kits.
If you really want to go to dissolvable resins then check Frank's work in: Gubler, F. (1989). Immunoflourescence localization of microtubules in plant root tips embedded in Butyl-Methyl Methacrylate. Cell Biol International Reports, Vol. 13, No. 1, January, 1989.
Eric Hines Microscopy Centre CSIRO Entomology Canberra
Have you tried to do regular gold followed by silver enhancement or gold toning on the semi-thin LR White sections? Then you'd even be using the same secondaries on your tests. I have no ideas on the fluorescence working (or not), but could you please post any responses to the server?
A few weeks ago I posted a querry to the listserver wondering why I was not getting good fluorescent labeling using otherwise successful primary antibodies applied to the surface of one micron thick LR White sections followed by secondary TRITC or FITC conjugates. I sincerely thank those that responded, and I am now sharing the responses:
I have tried labelling 1 micron sections with lectins conjugated with TRITC and they worked exceptionally well.... that is the lectin was labelled with the TRITC tag...
Eric Rosen
I suspect the problem lies in the different secondaries, maybe try the fluorescent-gold-secondaries that are commercially available to test fluorescence and gold label at the same time. The product is called fluoronanogold and is available from Nanoprobes Inc. You can tap into their website (http://www.nanoprobes.com) to obtain detailed protocols or contact them at 516-444-8815. They also put out a newsletter.
Margaret Springett e-mail hukee.margaret-at-mayo.edu IEM Specialist at Mayo Foundation 1426 Guggenheim Rochester, Mn. 55905
I routinly test 1 micron LR White sections on glass before going to the trouble of Immuno EM. I usually use peroxidase ABC which seems to enhance the signal enough to be visualized. However, the fluorescent signal is very weak unless you start stacking antibodies which can create more backround. So you need a very good optimized scope to be able to see the signal and your faint signal may be obscured by the backround signal of the aquamount which has higher fluorescence than some others. Prolong from M olecular Probes is very quiet and enhances Texas Red. Or my favorite for quiet and quick is: 70% Glycerol, 25% .5M Tris pH 9, 5% n-Propylgallate, heat to mix then pH to 7.4 and store in the fridge. For use: 1 drop, swirl, drain most of it off then cove rslip. There should be barely enough to cover the coverslip.
Good Luck Doug
Robert Underwood Morphology Core Dermatology U of Wash. Seattle, WA
We had the same disappointment a few years back when we went from great IHC labelling at the EM level to nothing with half micron LR sections. We got round the problem in two ways. One, we gold labelled the half micron LR sections then silver intensified and photographed with white light. Two, we went back to embedding in wax, cut 5 to 10 micron sections, de-waxed and got excellent FITC-IHC labelling. Although we were reticent to go to wax embedding (19th century technology) it rendered available such enormous areas of antigenic sites that the IHC was excellent plus we could use the sections for in situ work and the fluorescent apoptosis kits.
If you really want to go to dissolvable resins then check Frank's work in: Gubler, F. (1989). Immunoflourescence localization of microtubules in plant root tips embedded in Butyl-Methyl Methacrylate. Cell Biol International Reports, Vol. 13, No. 1, January, 1989.
Eric Hines Microscopy Centre CSIRO Entomology Canberra
Have you tried to do regular gold followed by silver enhancement or gold toning on the semi-thin LR White sections? Then you'd even be using the same secondaries on your tests. I have no ideas on the fluorescence working (or not), but could you please post any responses to the server?
A "RFQ" for products which, to the best of our knowledge are similar to those offered by you ,was placed with us by one of our clients.
We are a world wide sourcing firm and we are paid by our clients to find them suitable suppliers .
To you ,our service is totally free of charge !!!
The information we will get from you will not only be immediately sent to this particular client but also to all other clients looking for similar products .
To define and advise us the products you are interested to export and/or to get more information about us and our FREE SERVICE , please use our Internet interface at: http://www.thebol.com
Best Regards
N. Nissimoff Director theBOL - Purchasing Department
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } Salut to the listmembers,
Where to get freeware Wang Image (as *.zip files ?), on which URL address ? J o z e f Stankovic
} You might try Wang Image (now a standard free imager that comes with } windows 95, or is availiable as freeware I believe) We use it to convert } JEOL tiffs to tiff formats that Lview/NIH can read. It seems to read } many different tiff formats but only saves in one (which I don't know, } but a tiff saved by wang image suddenly becomes readable by all of our } other viewer programs).
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Steve Stokowski, I will try to send you single file (*.spe or *.lab) by e-mail and you`l to change the spectrum image to jpg or gif format, O.K. ? To send you file better by e-mail or FTP ?
Yours sincerely J o z e f Stankovic
} In a message dated 97-10-23 03:19:53 EDT, Kees Jalink wrote: } } { { Does anybody know a windows program that can read IPlab's TIFF format (12 } bit greyscale, stored in 2 bytes, I think) and convert it to any } windows-recognized format? } } } } PaintShopPro, ver. 4.14, from JASC will read 6 different TIFF subformats. } These are: uncompressed, Huffman compresssed, pack bits compressed, LZW } compressed, Fax Group 3 compressed, and Fax Group 4 compressed. None of } these are 12 bit grayscale. They range thru 1, 4, 8, and 24 bit/pixel color } scales. Attach a single file to an e-mail to me and I will try to change } the image to something else, such as a gif or a jpg, your choice. PSP } supports 35 different formats. } } Yours truly, } Steve Stokowski } Stone Products Consultants } Concrete Petrographers } 10 Clark Street, Suite A } Ashland, Massachusetts, 01721 USA } 508-881-6364 } http://members.aol.com/CrushStone/index.htm
Vachik Hacopian wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Charles Duvic of Ladd Research wrote: } } } Acetone and ethanol are viable alternatives to Propylene oxide. Ethanol } } should be used cautiously because it may inhibit epoxy polymerization. } } What is meant here by the word "cautiously"? Is it in reference to the } ethanol not being absolutely dry? } } Vachik Hacopian
Dr. Hacopian, Perhaps " cautiously" was too strong of word. I simply meant that care must be taken to ensure that all ethanol is removed from the speciman before any attempt at polymerization. Some investigaters have have noted that a "small" amount of ethanol in the speciman doesn't seem to affect polymerization of the block. Since "small" is subjective and not clearly defined I think it is best to wash the speciman with a resin mixture until all (or most) ETOH is removed.
To Eriko: unfortunately it is not possible to communicate with you by direct = e-mail (see below). Eriko, you did=B4nt send with your postal adress! Please forward it to me Thank you very much Wolfgang MUSS ------------------------------------------------------------
----- Transcript of session follows ----- mail: Cannot append to /var/mail/terao
----- Message header follows ----- Received: from aesculap.lkasbg.gv.at by fisher.sc.ucl.ac.be = (5.x/SMI-SVR4) id AA15831; Mon, 13 Oct 1997 09:16:53 +0200 Received: (from smap-at-localhost) by aesculap.lkasbg.gv.at (8.6.12/8.6.9) = id IAA27090 for {terao-at-bani.ucl.ac.be} ; Mon, 13 Oct 1997 08:11:18 +0100 Received: from unknown(10.1.101.6) by aesculap.lkasbg.gv.at via smap = (V1.3) id sma027088; Mon Oct 13 08:11:03 1997 Received: from c1pa008.lkasbg.gv.at (c1pa008.lkasbg.gv.at [10.1.42.8]) by hermes.lkasbg.gv.at (8.8.5/8.8.5) with SMTP id LAA06414 for {terao-at-bani.ucl.ac.be} ; Mon, 13 Oct 1997 11:09:34 +0200 Received: by c1pa008.lkasbg.gv.at with Microsoft Mail id {01BCD7B8.54DF35C0-at-c1pa008.lkasbg.gv.at} ; Mon, 13 Oct 1997 09:13:55 = +-200 Message-Id: {01BCD7B8.54DF35C0-at-c1pa008.lkasbg.gv.at}
Hello,
after several unsuccessful attempts to stain PE ultrathin cuts with the Kanig method I would like to ask if somebody has been successful and is willing to share his protocol with me. The specimen is already cut with an cryo-ultramicrotome and the cuts are lying on copper grids.
TIA,
Petra -------------------------------------------------------------- Dr. Petra Wahlbring Centre de Recherche Public Centre Universitaire (CRP-CU) Laboratoire d'Analyse des Materiaux (LAM) 162a, av. de la Faiencerie L-1511 Luxembourg tel. +352-466644-402 fax +352-466644-400 e-mail: petra.wahlbring-at-crpcu.lu
We are exploring the use of ambient atomic force microscopy to study the surface of transition metal oxide. We would like to contact research groups who are involved in the simulation of water adsorption on transition metal oxide(v2o5,MoO3) Who are the contacts for this type of work?
} may-be this question doesn't belong here, but I am totally stuck so I } still ask it: } I have recently taken some 120 Mb of digitized pictures with a } CCD-camera equiped microscope using IPlab software on a MacIntosh. Does } anybody know a windows program that can read IPlab's TIFF format (12 bit } greyscale, stored in 2 bytes, I think) and convert it to any } windows-recognized format? These data are important for me. } Thanks in advance. } Kees Jalink } } -- } } } Kees Jalink } The Netherlands Cancer Institute, dept. of Cell Biology H1 } Plesmanlaan 121 1066CX Amsterdam, the Netherlands } 020-5121982 (tel) / 020-5121989 (fax) } kees-at-NKI.NL (email) / 0297-320248 (tel at home)
I'd do it the other way. Usa Graphic Converter, a widely available Mac shareware application, to do the convertion on the Mac. Graphic converter can read about 50 graphic formats and saves to about 40, including a range of PC formats. Not sure if it will read in your particular TIFF files, but if not, you could contact the author - he has added in a number of proprietary formats on request.
Regards,
-- Larry Stoter 17, Rocks Park Road, Uckfield, E. Sussex, TN22 2AT, UK email: LPS-at-teknesis.demon.co.uk Phone/Fax: +44 (0)1825 767967
Ian, As long as the tempature is between 10-15C the CO2 will be a liquid. I use water from a sink to cool my CPD. Even in the summer the water is cool enough to get liquid CO2. I can't help you with your other problem.
-- Greg Rudomen Greg-at-umic.sunysb.edu S.U.N.Y. Stony Brook University Microscopy Imaging Center 516-444-3126
IAN HALLETT wrote:
} } We have a Peltier cooled Critical Point Dryer (CPD750 - manufactured } by EMSCOPE) that is having problems attaining a low enough } temperature prior to flushing with carbon dioxide. The manual } suggests a temperature below 10C - we can rarely get to below 12C and } frequently only get to 14C. Ambient temperature at the moment is } only around 20C. Has anyone any suggestions? As usual we have no } specifications for the electronic circuit nor a circuit diagram. } }
I use Spurr's resin for general e.m. histology and LR White (London Resin Company Limited) for most other things. They can both be mixed with absolute alcohol and so I can avoid the problems with propylene oxide.
I know that Spurr's is more carcinogenic than other epoxides but at least it isn't as volatile as propylene oxide which also has an annoying habit of melting many types of gloves. But I would be interested to hear anyone else's opinion about which is nastier: Spurr's resin with alcohol or epon with propylene oxide.
Malcolm Haswell E.M. Unit University of Sunderland
Disclaimer - these are my opinions and not necessarily those of my employer.
----------
Norbert
I use Spurr's resin for general e.m. histology and LR White (London Resin Company Limited) for most other things. They can both be mixed with absolute alcohol and so I can avoid the problems with propylene oxide.
I know that Spurr's is more carcinogenic than other epoxides but at least it isn't as volatile as propylene oxide which also has an annoying habit of melting many types of gloves. But I would be interested to hear anyone else's opinion about which is nastier: Spurr's resin with alcohol or epon with propylene oxide.
Malcolm Haswell E.M. Unit University of Sunderland
Disclaimer - these are my opinions and not necessarily those of my employer.
----------
Peggy Brannigan wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } Hello all, } } I'd like to de-embed some LX112 embedded plant leaf tissue so that I could } examine it in the SEM but, never having done this before, I'm looking for } advice, tips, references, protocols etc. Also, is it possible to de-embed } thin sections already stained (uranyl acetate, lead citrate) and examined } in the TEM? (Assuming I could get them off the grid....) } } Thanks,
Peggy,
We at LADD sell LX-112 and one procedure we know of that may work for you is prepared as follows:
1. Prepare saturated solution of KOH in absolute ethanol. Let stand overnight. 2. Pour off supernatant fluid. This is the epoxy solvent. 3. Trim block of excess epoxy resin. 4. Let speciman soak in solvent until resin is removed. Time will vary depending on size of block. 5. Wash with several changes of absolute ethanol. 6. Prepare for SEM
NOTE This solution may do damage to your speciman. I suggest trying this procedure on only one or two of your less critical samples.
Good luck,
Dr. Charles Duvic Ladd Research tel 1-800-451-3406 fax 1-802-878-8074
Below is the result of your feedback form. It was submitted by (gusev-at-ipm.sci-nnov.ru) on Friday, October 24, 1997 at 07:05:18 ---------------------------------------------------------------------------
Email: gusev-at-ipm.sci-nnov.ru Name: Gusev S.A.
School: IPM RAS, Nyzhnii Novgorod University
Zip: 603600
Question: We have made periodical 2-D arrays of the magnetic single domain particles. The size of the each particle ~30 nm. We measured their cooperative properties and would like to see the state (the direction and the magnetude of magnetization) of the each particle. Who can help us to realize our dream by means of the electron microscopy?
I just wanted to thank you all for your help. I just replaced the Tonner cartridge in our Lexmark Optra, tidded up the printer, and the first test print out was free from spots, speckles, blemishes! We're greatly happy with our Lexmark once again.
Take home lesson - when printing any sort of images, even a lite quaitity mixed with mostly text, the 7K-pages and 15K-pages are not anywhere in the ball park (we started having problems -at-~2,500 pages and totally unacceptible at 3K pages with our 7k-page cartridge)
Thank you.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"640K ought to be enough for anybody." -- Bill Gates, 1981
As I was asked to put a summary on the server so here it is. Thank's again to every body who answered me.
Marc
Just take a small amount of the Soot and put it in a vial and add Acetone (10ml) and then ultrasonicate the solution for 3-4 mins. Then you need to place a few drops on a carbon coated grid, let the solvent evaporate and put the grid in the scope. First use low mag to locate the material on the grid. It is best to use 100-200 kV to image the tubes. KMJ
I sssm to recall that the powder can be suspended in a solvent (benzene or toluene-I don't remember which). A single drop onto a carbon coated copper mesh grid is sufficient.
Hi, I had the same problem, so I can recommend you to make a suspention of this powder in alcohol, then put a drop on a grid with deposited thin film. Contrast will be poor, so you will need to use defocused image.
Marc, There are many reports of TEM on carbon nanotubes. However, you can sonicate the powder containing the nanotubes in ethanol or water, then place a small aliquot on the TEM grid and wick away the excess liquid. It's important to make sure the carbon actually gets suspended in the liquid you are sonicating with. I use a pipette to help mix the suspension quickly.
We examine carbon nanotubes as follows:
1. suspend the specimen in a small volume of either acetone or methanol (trial and error will be needed to determine the exact dilution. so try different dilutions)
2. suspend the sample by swirling (some people sonicate for several minutes) and use a pipette to transfer a droplet onto a holey, filmed (Butvar or Formvar) and carbon coated grid (suggest purchasing from commercial source)
3. allow to air dry and examine in TEM looking for tubes suspended over the holes (best resolution)
The TEM should be set up for hi res, cooled traps over specimen area and a clean vacuum system is needed.
Good luck.
I used to view these regularly for a materials scientist. We simply put a small amount of the carbon sample in a solvent and used a sonicator to disperse it. Ten minutes should do a decent job. We then put drops of the suspension onto holey grids which were sitting on filter paper. When they were dry we put them in the TEM for viewing. Some of the nanotubes would sit on the edges of the holes in the film and could be viewed that way.
The nanotubes we viewed required very high magnification and very stable beam and vacuum conditions. Use liquid nitrogen on the pumps and on your decontaminator. Set up your instrument for high resolution conditions and let the electronics and gun stabilize for a while before viewing. Getting the best resolution was trickier by far than preparing the samples.
We just make a dispersion of the powder ( say in methanol) and put a drop on a holey grid and we let it dry for a few minutes. We examine the nano tubes at about 100K to 300K. If you use regular carbon coated grids ( rather than holey grids) the image of the nano tubes will not be as clear because the carbon background will interfere.
------------------------------ SCHMUTZ Marc IGBMC 1 rue Laurent FRIES BP 163 F 67404 Illkirch Cedex FRANCE
} as many of you will be aware, it can be very difficult to accurately } determine diffraction ring diameters in selected area diffraction patterns } recorded from multicrystalline TEM specimens. Especially when rings are } very spotty.
This is true even if "signatures" between patterns are obviously different by eye, at least for large unit cells. Before the days of digitization, we developed a method for making sense of such patterns from mixed-mineral specimens (esp. interplanetary dust particles collected in the earth's stratosphere) which was published in Micron around 1980. Let me know if you would like the reference.
} What I'd like to do is digitize the SAD pattern, locate the exact centre } and intergrate pixel intensities through 180 degrees. This will result in a } one dimensional diffraction pattern with pairs of spots either side of the } undiffracted spot, which should be much simpler to measure.
One of our attempts in those early days involved "photographic" azimuthal averaging: putting film on a turntable whose center aligned with the projected pattern center, and then exposing the film for a couple revolutions. I don't recommend this at all!
} I would like to know if anyone out there has a computer program to do this } type of SAD pattern manipulation. I propose to use Photoshop and a flat } bed scanner on a Macintosh to digitize the SAD patterns which would be } saved as TIFF or some other suitable format. So I suppose the ideal } solution to my problem would be a Photoshop plugin. I also use NIH Image } so a plugin for this program would also be suitable.
The image processing language Semper has verbs for both locating the pattern center very precisely, and for azimuthal averaging while treating images not simply as bytes but as (real or complex) floating point numbers. If you get access to a Semper interpreter (my contact address for the company keeps getting lost), I'll be happy to share our macros with you.
} I would really appreciate any help with a plugin, stand alone program (Mac } or PC) or comments on how I should go about writing my own solution. I } look forward to your replies,
We've written a Visual Basic program for doing this, but I'm not sure it's sufficiently error-trapped for outside distribution. If Java programs could read local system data files on the request of a local user (can they?), I could probably whip together a program that everyone could use in a couple of hours.
In a message dated 97-10-24 02:38:53 EDT, you write:
{ { I will try to send you single file (*.spe or *.lab) by e-mail and you`l to change the spectrum image to jpg or gif format, O.K. ? To send you file better by e-mail or FTP ? } }
Dr. J o z e f Stankovic:
I do not know what to do with a .spe or .lab format. Sorry. Maybe somebody on the Listserver can help.
I have been a happy user of this system for about 5 years(just upgraded to the WIN 95 version). The answer to all of your question but one is yes it can do that.. I don't know about the vidio conversion though.. But than again I use Adobe for that. On Tue, 21 Oct 1997 kszaruba-at-MMM.COM wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear Listers, } } I've heard a few mentions of the BioQuant system in the past. } Now I am considering it myself, and would like to hear a little } more detail from users. } } What do you like most/least about the system? How easy is it to } handle intensity comparisons of stained specimens (BF and Fluor)? } What camera option did you choose and how well does it work with } a variety of specimen types (macro copy stand, micro BF, fluor, } moving subject, etc)?? Does it really do excellent gel } densitometry? How about colony counting? 3D measurements? Can } it turn VCR tapes (short) into digital movies? } } I'd appreciate any comments! } Thanks as always, } Karen } } P.S. Regarding the Message Subject requirements, what about } general subjects like digital imaging, printers, etc?? LM vs. } TEM, etc. doesn't really fit. } } -- } Karen Zaruba } kszaruba-at-mmm.com } 3M Company, 3M Center Bldg. 270-1S-01 } St. Paul, MN 55144 } "The opinions stated above are my own, not necessarily 3M's" }
The file is about 2.0 MB and will expand to 2.2 MB after decompression.
At 08:10 AM 10/24/97 +0000, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The November meeting of the Mic. Soc. of Northeast Ohio will welcome S. Frank Platek, of the FDA - Forensic Chemistry Center (Cincinnati, OH) as our featured speaker on Thursday, November 6, 1997. Details are as follows:
Title: "Forensic Microscopy Related to Product Tampering and Counterfeiting" The evening's presentation will 'focus' on the Forensic Chemistry Center's multiple microscopy investigation of product tampering and counterfeiting of foods, beverages, pharmaceuticals, and medical devices. Specifically, the use of stereoscopic, polarizing and comparison light microscopy, as well as scanning electron microscopy, and energy dispersive x-ray analysis in State and Federal cases. Techniques for case analysis and some uncommon used of image analysis; backscattered electron imaging and x-ray mapping will be featured along with case histories.
Place: Case Western Reserve University (Medical School) Andrews Conference Center, room 6306
Time: 5:45 Reception/Social Hour 6:30 Presentation 8:00 Dinner at Club Isabella Menu includes: Soup or salad; choice of Grilled Salmon and Pasta Primavera; OR Herb Roasted Chicken Breast; OR Pasta Sauteed with Fresh Spinach. Dessert Tray and Coffee for $20.00 (make checks payable to MSNO).
Meal choices and reservations must be made by noon, Monday November 3, 1997 to Valerie Woodward, MSNO Secretary (216)447-5408 (voice) or woodward-at-brk.bfg.com (E-mail).
does anyone know where I can find a source for the laser that fit on the Gatan Duo mill, and are connected with the auto-terminator with a bnc connector? Any hints are welcome: maker, vendor, etc.
I would be grateful if you would bring the position below to the attention of potential candidates.
Many thanks,
Steve Pennycook
{fontfamily} {param} Times {/param} {bigger} POSTDOCTORAL POSITION IN MATERIALS PHYSICS
OAK RIDGE NATIONAL LABORATORY, SOLID STATE DIVISION
A postdoctoral position is available in the Electron Microscopy Group led by Dr. Stephen J. Pennycook at ORNL, in collaboration with Prof. Elizabeth Dickey of the University of Kentucky. Although the position will be joint between ORNL and the University of Kentucky, the post-doctoral researcher will be on permanent assignment at ORNL. The research programs to be pursued in this position include: (1) Structure and Chemistry of Novel Nanotube Materials and (2) Interface Structure and Bonding in High-Temperature Oxide Composites. In the nanotube project, materials with novel physical properties are being developed at the University of Kentucky through selectively doping and functionalizing single wall nanotube (SWNT) bundles. Critical to the project is a fundamental understanding of the dopant distribution in the SWNT bundles and the subsequent effect on structure and electronic properties. In the oxide composite program, we seek to understand the atomic structure and chemistry of interfaces in these high-temperature structural materials and to develop correlations between the atomic structure and mechanical behavior of the interfaces. Both projects will require complementary atomic-scale electron imaging and electron energy loss spectroscopy (EELS), so the successful candidate should have practical experience in both techniques.
The Solid State Division has some of the world's finest facilities for atomic scale imaging of materials: a VG Microscopes HB603 300 kV scanning transmission electron microscope with a 1.26=C5 probe size provides direct, Z-contrast imaging capabilities for interfaces in materials. A VG Microscopes HB501UX 100 kV microscope with a high sensitivity parallel EELS capability provides atomic resolution spectroscopy. The Electron Microscopy Group has two Silicon Graphics=20 workstations with the Molecular Simulations' Cerius 2 package incorporating the CASTEP pseudopotential code. The Division has a number of additional workstations, and access to extensive parallel computing capabilities, including the Intel Paragon XP/S 35 and XP/S 150 with 512 and 2048 processors respectively.
I noticed that you mentioned SPE and LAB files which I recognize as being from a Link system. We also have one. Those files will not convert readily since they are a proprietary format. But we have a couple of approaches that we use to embed Link spectra in a document.
First, the file may be pasted into a document with the following procedure. - Prepare the spectrum for printing and begin the print dialog. The print preview window will display. - From the EDIT pull down menu select COPY. - Switch to the application you wish to paste into. - From the EDIT pulldown menu select COPY SPECIAL and paste the spectrum in as a picture (not as text which copies in only the header information). - The spectrum will appear as a resizable graphic line drawing with spectrum and axis labels.
Second, a snapshot of the spectrum window may be processed. - Prepare the spectrum for printing, but stop short of starting the printing process. - Activate the zoomed spectrum or main spectrum window (your choice), and press Alt-Prtscrn. This will copy a bitmap of the active window just as it appears to the clipboard. - Switch to the application of choice such as Word or MS-Imager (I like Imager). Paste the clipboard contents, or use the File New From-Clipboard function in Imager. The new image may be cropped to eliminate the unwanted features.
This second procedure works with any Windows (3.x or 95) application window. Hope this help.
At 10:29 AM 10/25/97 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Some of the user's here are embedding plant tissue into LR White after doing an acetone dehydration (I know it's not the recommended dehydrating solution, so don't nag me on that = ) ). My question is: Is there a stain that is soluble in acetone that we could use in our last 100% acetone step that would stain the plant tissue so it is visible for embedding and sectioning. When we use ethanol for dehydrating we use a 0.1% safranin O and it turns the plants a lovely pink without affecting the immunostaining capacity. Is there a stain out there that would work in acetone & do the same thing?
If anybody out there knows of one....
I'm dyeing to hear about it.
Freeing the radicals (acetone, that is) in Berkeley,
Paula = )
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
Dear All: I am forwarding this request for a donation of an oil immersion microscope from my son, Rene Jordan, who is a volunteer working for the Peace Corps in the Philippines. The microscope is needed for the detection of Malaria. I hope this is not un unreasonable solicitation, I am not familiar at all with the value of such a microscope. Abount my son: He graduated with a degree in environmental science from UCI, Calif. and joined the Peace Corps in April. After his 3 months training in Manila he was sent to one of the 7000 islands for coastal resource management, mainly sea turtles and coral reefs. The small village he is in has no phone (how nice), no paved roads and every night at 9 the generator is turned off except Tuesdays were it runs till 10 for the church service. To get his mail he travels once a month for 6 hours to the next bigger city. If you have one of those microscopes standing around please let me know or you can contact him direct: Rene Jordan - PCV PCSDS 3/F Capitol Complex Puerto Princesa City, 5300 Palawan, Philippines Thank you very much, Peter Jordan
Hello, I use eosin B (Kodak cat# 117 7930) in 100% ethanol to stain the tissue before embedding in LRWhite for immunostaining. So far I have never had any problems with the immo-detection of various antigens . Eosin B can be dissolved in acetone as well.. I use at 1%..hope it helps Neelima Shah. Morphology core, UOP
} Boarders, } } =A0=A0=A0=A0=A0=A0=A0 Some of the user's here are embedding plant tissue= into LR White } after doing an acetone dehydration (I know it's not the recommended } dehydrating solution, so don't nag me on that =3D )=A0 ).=A0 My question= is:=A0 Is } there a stain that is soluble in acetone that we could use in our last 100% } acetone step that would stain the plant tissue so it is visible for } embedding and sectioning.=A0 When we use ethanol for dehydrating we use a } 0.1% safranin O and it turns the plants a lovely pink without affecting the } immunostaining capacity.=A0 Is there a stain out there that would work in } acetone & do the same thing? } } =A0=A0=A0=A0=A0=A0=A0 If anybody out there knows of one.... } } =A0=A0=A0=A0=A0=A0=A0 I'm dyeing to hear about it. } } } Freeing the radicals (acetone, that is) in Berkeley, } } } Paula=A0=A0 =3D ) } } Paula Sicurello } UC Berkeley } Electron Microscope Lab } psic-at-uclink4.berkeley.edu=20
} Thanks a lot to all who gave me some nice tricks to observe carbon } nanotubes. It helped me a lot and due to your nice indications I've got } correct images at the first shot.
} Marc ---------------------------------------------response------------------------- ----------------- Marc:
If you have time and the inclination I would be interested to see a brief description of the technique you finally used.
I have a question which has undoubtedly been answered before...! We are going to buy a framegrabber and a camera. We use a Power Mac 8200 and want to replace the old greyscale framegrabber with a color one. The camera is used in order to capture images of plants under development, but also for capturing images of cells under a microscope. Should we go for digital cameras? What is the difference between an AV card and a framegrabber?
Can somebody help us in order to take the right decision?
Thanks for all the suggestions and help for tranferring IPlabs 12-bit grey-scale TIFF format (MacIntosh) to windows. I tested some 20 different commercial and freeware viewers, converters, etc. In my hands, the only one that worked without any problems is Thumbsplus (ver3.10) by Philip Crews (Cerious software; available as shareware). Just copy the files to a PC directory, rename them to *.tif, and Thumbsplus will read them, correctly noting that it is 16-bit greyscale (last 4 bits not used) and motorola byte order. Nice piece of software for 65$.
((Salut to the listmembers,
may-be this question doesn't belong here, but I am totally stuck so I still ask it: I have recently taken some 120 Mb of digitized pictures with a CCD-camera equiped microscope using IPlab software on a MacIntosh. Does anybody know a windows program that can read IPlab's TIFF format (12 bit
greyscale, stored in 2 bytes, I think) and convert it to any windows-recognized format? These data are important for me.))
Kees Jalink The Netherlands Cancer Institute, dept. of Cell Biology H1 Plesmanlaan 121 1066CX Amsterdam, the Netherlands 020-5121982 (tel) / 020-5121989 (fax) kees-at-NKI.NL (email) / 0297-320248 (tel at home)
Inside IPLab spectrum you can change the data types from 16 bit to 8 bit images. It is under the math menu, change data types. Once you get it to 8 bit, the images should be readable in any Mac or PC program that can open tiff files.
One other note, I use MacLinkPlus to transfer tiff images between PC and Mac and visa versa.
Hope this helps, Lou Ross
Senior Electron Microscope Specialist 101 Geological Sciences Building University of Missouri Columbia, MO 65211 (573) 882-4777, 882=5458 (fax) geosclmr-at-showme.missouri.edu www.missouri.edu/~geosclmr/ebaf.html
Hello Microscopy world! We have been asked to attempt to microscopically evaluate collagen films. The sheets are approximately 50% hydrated and are approximately 500 um thin. The client wishes to see if there are differences evident in the cross-linking that occurs as the film sheets are produced under various conditions (varying concentrations of constituents). It is necessary to evaluate them as close to their natural state as possible. Any suggestions? Thanks for accepting this challenge! Tracey Pepper Bessey Microscopy Facility Iowa State University
We would like to find a computer program to determine the misorientation that exists across a grain boundary from electron diffraction patterns. Specifically, if the electron diffraction patterns of adjacent grains were obtained, does a program exist which, by using the two ED patterns, would describe the grain boundary geometry by determining the grain boundary plane and rotation axis that exists between the two grains? We are interested in performing this analysis on non-cubic materials which possess a hexagonal or rhombohedral structure.
Thanks in advance.
Owen
============================= Owen P. Mills Michigan Technological University Metallurgical & Materials Engineering Rm 512 MME Building Houghton, MI 49931 906-487-2002 906-487-2934 FAX opmills-at-mtu.edu
Hello Microscopy World! We have been asked to microscopically evaluate thin sheets of collagen films. They are approximately 50% hydrated and about 500 um thin. Our client wishes to see the differences in the cross-linking of the collagen produced under various conditions. It is necessary to evaluate them under as natural conditions as possible. Any suggestions? Thanks! Tracey Pepper Bessey Microscopy Facility Iowa State University
Software to calculate misorientation from Kikuchi patterns is quite common, however typically homemade (I used to have one written in FLEXTRAN, running on Tracor Northern analyzers, but that is history). I enclose below a list of those microscopist which I know having written such programs recently. Stefan and Robert both have software which could already have build in your application to determine grain boundary indices (you need to determine the direction of the grain boundary with respect to the orientation of the unit cell of the grain of interest as well the inclination of the grain boundary in the foil, typically obtained by a tilt analysis). Stefan also has a routine build in which is very much useful for Burgers vector analysis. All programs are suited for on-line analysis and are PC based.
Stefan Zaefferer: stefan-at-isma.u-psud.fr Robert Schwarzer: robert.schwarzer-at-tu-clausthal.de Paul Baggethun: baggeth-at-sms.emse.fr
Hasso Weiland Alcoa Technical Center Alcoa Center, PA 15069
} ---------- } From: Owen P. Mills[SMTP:opmills-at-mtu.edu] } Sent: Monday, October 27, 1997 10:19 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM - Electron diffraction software } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am interested in getting some feed back on Digital Capture packages from various vendors. I specificaly want to place system on a JEOL 35C SEM. I aminterested in knowing how well the various vendors perform specificaly SemiCaps. If anyone has any information such as pros and cons to any one particular system I would realy like to hear it. Please send emails to rgarcia-at-cs.wright.edu. Thanks.
Roberto Garcia Electron Microscopy Facility Manager Wright State University
Thank you to those who responded so far to my question on reduction of OsO4 and RuO4 to less hazardous states using corn oil.
So far no one has responded with experience with RuO4 and corn oil - any others lurking out there want to add 2 cents? I did get a very detailed reply from Wolfgang Muss on using ethanol to inactivate OsO4. Also I have never heard of the "reprocessing hazardous waste" issue that Steven Barlow brought up. Sounds like a wonderfully forward-thinking, environmentally responsible policy to me. [Must have been designed by the same folks that allow liability issues re:children to make it too costly for our neighborhood to have a park.] Does this reprocessing issue apply outside San Diego?
Answers to two questions: The corn oil procedure for OsO4 appears in a "Technical Data Sheet" from Electron Microscopy Sciences, with the following references given: (I haven't read them, just the data sheet)
Cooper, K. Neutralization of Osmium Tetroxide in case of accidental spillage and for disposal. Bulletin of The Microscopical Society of Canada. 1988. 8:24-28. (Also I think I saw similar article in Don Grimes' free publication, Microscopy Today).
Lunn, G.;Sansone, E.B. Osmium Tetroxide. Destruction of Hazardous Chemicals in the Laboratory; Program Resources, Inc. Frederick, MD; p211-213.
The other question was about the hazards of Sodium Bisulfite. I am just going by an MSDS I have for CAS # 7631-90-5 from Sigma-Aldrich. This states that sodium bisulfite is Toxic, Harmful by Inhalation and Contact, causes Severe Irritation, and is a Possible Sensitizer. Taking into account the fact that even bread pudding would probably show up as harmful by inhalation on an MSDS, still the designations of Toxic, Sensitizer and "Severe" Irritant are worth noting.
Still appreciate any further comments! Karen
-- Karen Zaruba kszaruba-at-mmm.com 3M Company, 3M Center Bldg. 270-1S-01 St. Paul, MN 55144 "The opinions stated above are my own, not necessarily 3M's"
I am a post doc working in nanotechnology at Washington University in St. Louis. I have a general question regarding calibration standards for microscopes. We are researching possible uses of a collections of small monodispersed particles and it occurred to us that there might be a use for them in calibrating microscope systems.
Specifically, I am interested in scanning probe microscopy (AFM,STM,MFM), electron microscopy (SEM,TEM) and light microscopy (both optical and near-field optical scanning microscopy).
What sort of small particles are used (if any) to calibrate these instruments? We are interested in the composition and size.
If collections of small particles are not generally used, would there be an interest in the microscopy community for a collection of small particles (~100 nm to 10 microns) to be used as a calibration or any other sort of aid.
I would be very interested in your reply,
Thanks very much
-Stephen Irons
********************** Washington University Department of Physics CB1105, 1 Brookings Drive St. Louis, MO 63130
Iam an international student accepted in the biology department at New York University. I have received my Master's degree in microbiology. I would like to know more about your scholarship for graduates. Thank you
I am experiencing an excessive charging in my SEM. The tested samples are conductive, the specimen mount, holder and SEM stage are all well grounded, but charging persists, precluding succesful imaging. Secondary and backscattered electron images are equally affected. These samples do not exhibit charging when analyzed under the same conditions (AV) in another SEM. Many different samples were tested in these two SEMs and the results are consistent.
It appears that some instrumental factor is involved. I would appreciate the input on the subject. Thank you very much in advance.
Chris Terlecki Applied Analytical Sciences ph: 714-434-6894 email: aas-at-pacbell.net
Dear Karen, } } So far no one has responded with experience with RuO4 and corn oil - any } others lurking out there want to add 2 cents?
Just a guess, but since Ru is also a group VIII metal, and since Ruthenium red is used to stain the lipid components of membranes, an un- saturated oil should combine with Ru in much the same way as with Os.
} Taking } into account the fact that even bread pudding would probably show up } as harmful by inhalation on an MSDS, ...
As the recent discussion of the MSDS of H2O would indicate. Yours, Bill Tivol
I've been asked to find out what software people are using to simulate electron diffraction patterns, specifically selected area diffraction patterns. We would like to know the name of each software package, supplier, approximate price, computer platform required, etc. Your comments on accuracy and ease of use would be most welcome.
We would also like to hear from people who have had bad experiences with commercial software. But please contact me directly, not via the listserver. We don't want to upset anyone unnecessarily.
Thankyou,
Mark Blackford TEM Group Materials Division, ANSTO PMB 1, Menai, N.S.W. Australia 2234 Phone 61 2 9717 3027 Fax 61 2 9543 7179
Disclaimer: The views expressed in this E-mail message do not necessarily represent the official views of ANSTO from which this message was conveyed.
I am trying to view air samples captured on a teflon filter with a Cambridge SEM. Is there a standard procedure that I should follow?
Joe Wise Director, W. M. Keck Math/Science Institute Crossroads School for Arts and Sciences 1714 Twenty-First Street Santa Monica, CA 90404 (310) 829-7391
Stephen Irons wrote: ============================================== I have a general question regarding calibration standards for microscopes. We are researching possible uses of a collections of small monodispersed particles and it occurred to us that there might be a use for them in calibrating microscope systems. ============================================== You might want to consider the Mag*I*Cal TEM Calibration sample, and unlike polymeric calibrated microspheres, there is nothing that is "changed" by the electron beam. Full details about the Mag*I*Cal sample can be found on our website given below.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
} Date: Mon, 27 Oct 1997 21:08:35 -0800 } To: jwise-at-kmsi.org (Joe Wise) } From: Mary Mager {mager-at-unixg.ubc.ca} } Subject: Re: air samples } } Dear Joe, } I have done exactly that on 60 different air filtered samples. My only comment would be to use Nucleopore-type filters (polycarbonate) if possible, because they are much smoother and show the particles up better than Teflon. The Teflon filters are quite rough and do not carbon coat well, so particles are harder to see and charging is a problem. Just cut out a small piece, attach to a stub, carbon coat and observe. } } } I am trying to view air samples captured on a teflon filter with a Cambridge } } SEM. Is there a standard procedure that I should follow? } } } } Joe Wise } } Director, W. M. Keck Math/Science Institute } } Crossroads School for Arts and Sciences } } 1714 Twenty-First Street } } Santa Monica, CA 90404 } } (310) 829-7391 } } } } e-mail: jwise-at-kmsi.org } } http://www.kmsi.org } } } Regards, } Mary }
Chris - and other microscopists - If indeed the specimen and stage are grounded, you should check that you are not using excessive beam current. Also, a large condenser spot or a huge final aperture can be the problem. Since BS too is affected grounding of the scintillator cannot be the problem. If a bare specimen mount does not charge, then the specimen itself is likely the problem. The other SEM may use just a little less beam current and a partially conducting specimen would then not charge up. If the problem is specimen related, one of the more common problems would be the umbrella effect; where the specimen is well coated on the upper surfaces, but these surfaces prevent a continuos good coating on the under sides. Visualise a stack of cannon balls or a mushroom: Coating from above would not be effective. Such specimens can be sputter coated by laying the pin type mount on the side, coating and then turning the specimen on the opposite side of the pine and giving it a second coating. At least this is a more interesting problem then fiddling with digital files. Oh, did you use that address "experts" deliberately? I understand the word means 'squirt under pressure'. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au ----------
} Dear SEM experts, } } I am experiencing an excessive charging in my SEM. The tested samples } are conductive, the specimen mount, holder and SEM stage are all well } grounded, but charging persists, precluding succesful imaging. Secondary } and backscattered electron images are equally affected. These samples } do not exhibit charging when analyzed under the same conditions (AV) in } another SEM. Many different samples were tested in these two SEMs and } the results are consistent. } } It appears that some instrumental factor is involved. I would } appreciate the input on the subject. Thank you very much in advance. } } } Chris Terlecki } Applied Analytical Sciences } ph: 714-434-6894 } email: aas-at-pacbell.net
} I am experiencing an excessive charging in my SEM. The tested samples } are conductive, the specimen mount, holder and SEM stage are all well } grounded, but charging persists, precluding succesful imaging. Secondary } and backscattered electron images are equally affected. These samples } do not exhibit charging when analyzed under the same conditions (AV) in } another SEM. Many different samples were tested in these two SEMs and } the results are consistent. } } It appears that some instrumental factor is involved. I would } appreciate the input on the subject. Thank you very much in advance. } } } Chris Terlecki } Applied Analytical Sciences } ph: 714-434-6894 } email: aas-at-pacbell.net
My first reaction is that it isn't charging if you see the effect is BSE images. You first need to really determine if it is the specimen or SEM. Same effect with entirely different specimens? If you move the same specimen between the different SEMs, is it always only in the one SEM? Use scan rotation - does the effect stay with the specimen or move with the scan direction. I guess I'd suspect some fault in the scan generation if it is really to do with the SEM - electrical fault in scan generation, or fault in coils, or connection to coils?
Regards,
-- Larry Stoter 17, Rocks Park Road, Uckfield, E. Sussex, TN22 2AT, UK email: LPS-at-teknesis.demon.co.uk Phone/Fax: +44 (0)1825 767967
We are interested in cryosections of plant material especially for elemental analysis with EDX. I would like to get informations about the necessary equipment (cryostat or ultracryomicrotomes) and experiences. Thanks in advance
Heike Buecking Dr. Heike Buecking University of Bremen UFT Plant Physiology and Plant Anatomy Leobener Str. D 28359 Bremen Germany TEL: +49-421-218-2954 or TEL: +49-421-218-7283 FAX: +49-421-218-3737 e-mail: heibueck-at-uft.uni-bremen.de FAX: +49-421-218-3737 e-mail: heibueck-at-uft.uni-bremen.de
I presume that for some reason you cannot coat the sample even though they are conductive. A wiff ie 5nm AuPd on the surface should overcome the charging. Also, turn down the wick on your microscope ie low kV, low beam current.
Patrick Echlin Cambridge
On Mon, 27 Oct 1997, Janusz Chris Terlecki wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear SEM experts, } } I am experiencing an excessive charging in my SEM. The tested samples } are conductive, the specimen mount, holder and SEM stage are all well } grounded, but charging persists, precluding succesful imaging. Secondary } and backscattered electron images are equally affected. These samples } do not exhibit charging when analyzed under the same conditions (AV) in } another SEM. Many different samples were tested in these two SEMs and } the results are consistent. } } It appears that some instrumental factor is involved. I would } appreciate the input on the subject. Thank you very much in advance. } } } Chris Terlecki } Applied Analytical Sciences } ph: 714-434-6894 } email: aas-at-pacbell.net }
It's not clear from your message what you are looking at. Are you really looking at air in which case use a helium cold stage and condense the stuff to a solid. If you are looking at stuff suspended in air see Chap 11 in "SEM & XRMA' Goldstein et al Plenum 1992.
Good luck.
Patrick Echlin Cambridge UKOn Mon, 27 Oct 1997, Joe Wise wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I am trying to view air samples captured on a teflon filter with a Cambridge } SEM. Is there a standard procedure that I should follow? } } Joe Wise } Director, W. M. Keck Math/Science Institute } Crossroads School for Arts and Sciences } 1714 Twenty-First Street } Santa Monica, CA 90404 } (310) 829-7391 } } e-mail: jwise-at-kmsi.org } http://www.kmsi.org } }
I have a question regarding what polymer resin to use for embedding a sample for (TEM) observation. The sample is a powder, which is likely to be moisture and CO2 sensitive. I am thus concerned that a condensation polymer would induce changes in the sample. We have been grinding the sample to produce a suspension for TEM specimen preparation. However, we would like to use ion milling, as this will be more likely to preserve the microstructure.
Does anyone know of a good embedding polymer for a moisture and CO2 sensitive sample, which is also high-vacuum compatible?
Wharton
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Wharton Sinkler PhD Department of Materials Science and Engineering Northwestern University 2225 North Campus Drive Evanston, IL 60208-3108 tel: (847) 491-7809 fax: (847) 491-7820 email: sinkler-at-apollo.numis.nwu.edu
Look for a grounding wire that may be disconnected. If your instrument capable of measuring the absrobed current, see if that is operating normally. A disconnected wire to ground from the stage will cause any sample to charge. Check continuity between the stage and the microscope body. Move the stage around while checking; you may have an intermitten contact.
-Scott Walck ----------
I would like to observe specimens on TEM substrates before and after ashing.
Specifically what I am trying to do is place a specimen on a suitable TEM substrate. Image the specimen and then image the specimen after ashing the specimen on the TEM substrate.
So far I have not been successful. I have tried germanium substrates on nickel and copper grids substrates. I also have tried gold on gold. The substrates decompose in all cases. I am surprised that the gold on gold decomposed.
I would be appreciated of any information related to this effort.
Regards,
Larry Murphy Group Leader, Analytical Section Cabot Corporation
I went through the MSA pages on the internet but could not get any information regarding the scholarships for graduate students. Please give a phone number or address or E-mail address to whom I can contact. thank you
I would like to put forward the presence of cellulose (or other major algal/Phaeophyceae/ cell wall components) in biological samples using fluorescence microscopy. I have heard about the existence of specific fluorochromes binding to cellulose. Could anyone give me more information about this.
If none of the other suggestions cure the problem, check for column contamination. A small bit of insulating material at the final lens or in the column can deflect (often erratically) the incident beam.
Woody White, Electron Microscopist SEM/EDS/WDS
Work: Mcdermott Technology, Inc. woody.n.white-at-mcdermott.com http://www.mtiresearch.com/
I have always used Calcofluor white M2R to visualize cell walls. I'm pretty sure it is specific for cellulose (I did find it's specificity described in writing, problem is that I was the author) but you might want to check.
================= C. John Runions Section of Ecology and Systematics Corson Hall Cornell University Ithaca, New York USA 14853
} I have a question regarding what polymer resin to use for embedding a } sample for (TEM) observation. The sample is a powder, which is likely to } be moisture and CO2 sensitive. I am thus concerned that a condensation } polymer would induce changes in the sample. We have been grinding the } sample to produce a suspension for TEM specimen preparation. However, we } would like to use ion milling, as this will be more likely to preserve } the microstructure. } } Does anyone know of a good embedding polymer for a moisture and CO2 } sensitive sample, which is also high-vacuum compatible?
} Wharton -
You don't say what TYPE of powder you're dealing with, but I'm wondering why you "would like to use ion milling, as this will be more likely to preserve the microstructure". Have you considered ultramicrotomy? There are standard embedding protocols, but selection of the right one depends on the sample.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
I believe that you better use ultramicrotomy to prepare powder samples for TEM. Particularly your sample is sentitive to drying and CO2. First, you embed your powder into resin which leads to completely enclose your powders from the environment. Then use Ultramcrotomy to cut thin-sections. I believe you will finds some places to prepare this type of TEM samples.
Larry Murphy wrote: ================================================== I would like to observe specimens on TEM substrates before and after ashing.
Specifically what I am trying to do is place a specimen on a suitable TEM substrate. Image the specimen and then image the specimen after ashing the specimen on the TEM substrate.
So far I have not been successful. I have tried germanium substrates on nickel and copper grids substrates. I also have tried gold on gold. The substrates decompose in all cases. I am surprised that the gold on gold decomposed. ===================================================== A technique has been developed that takes advantage of a SiO2 filmed grid followed by oxygen plasma etching. We at SPI, have used this method since the mid-1970's (credit for discovering it, so far as I know, is Dr. John T. Stasny who was working for SPI at the time) and it is useful for both materials science as well as life science thin sectioned samples:
1] Thin section the sample using normal diamond knife techniques but pick up the sections on SiO2 coated grids. 2] Do your pre-ashing TEM examination; the SiO2 film structure will not appear greatly different from what you are used to seeing with carbon. 3] Then place the grid in an oxygen plasma etcher and using pure oxygen, back fill to as low as possible a vacuum, and then "etch" for 10-20 seconds which should be enough for removal of organics in a thin section. The reason for the maximum vacuum for the backfill so to have the highest possible partial pressure of oxygen, thereby resulting in the fastest possible etching time. The oxygen plasma of course will not etch the SiO2 film. Of course, one does need a good stable SiO2 film. 4] You should be able to put the grid back into the TEM and photograph the same identical area after etching.
Tell me how it comes out. The plasma etcher (isotropic) being used should be operated at not more than 100 watts, other wise too much heat is generated. If you have a leaky system, letting in nitrogen, not too much has to be let in to drastically reduce the etching rate. The SiO2 film is made by evaporation of SiO at 10 -5 torr or better. When you buy them commercially, they are much cheaper than the alternatives you said you (unsuccessfullly) tried previously.
Disclaimer: SPI Supplies has been producing stable SiO2 films for this particular application and also offer the SiO for anyone wanting to make their own support films. We also produce the Plasma Prep II Plasma Etcher so we would have an obvious interest in seeing more of this sort of work being done. See our website, given below, for more information.
Chuck
PS: In a few weeks we will put up on our website a nice life science example of this kind of etching on a SiO2 filmed grid.
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I'm translating a microscope manual from Japanese and stuck with description that goes like, "The image position (of the eyepiece?) is set at 10 mm from the abutting joint."
Can anyone decipher my lousy translation and tell me what it means and if my diction is right?
Thanks in advance.
Chiba Atsushi [(Mr.) -- *Chiba* is my surname] Voice: (+81) 010-045-9451
Dear all, we are trying to determine the orientation of rutile crystals (20 microns) in a jarosite matrix. When we tried beamrocking, the rutile disappeared, probably because the jarosite acted as a flux. Has anybody any experience of this problem or any ideas as to how we might overcome it?
TIA Eric
---------------------- Dr Eric E. Lachowski University of Aberdeen Department of Chemistry Meston Walk Old Aberdeen AB24 3UE +44 1224 272934 e.lachowski-at-abdn.ac.uk
} Greetings! } } I have a question regarding what polymer resin to use for embedding a } sample for (TEM) observation. The sample is a powder, which is likely = to } be moisture and CO2 sensitive. I am thus concerned that a condensation } polymer would induce changes in the sample. We have been grinding the } sample to produce a suspension for TEM specimen preparation. However, = we } would like to use ion milling, as this will be more likely to preserve } the microstructure. } } Does anyone know of a good embedding polymer for a moisture and CO2 } sensitive sample, which is also high-vacuum compatible? } } Wharton Wharton,
It=D5s an interesting question you have. All of the conventional epoxy r= esins (O.K. At least the ones that I=D5ve used) are anhydride cross-linked . T= he polyester polymerization that I recall involves the formation of a ester = bridge between the two carboxyl groups of the anhydride and an epoxy group at ea= ch site. Thus, the reaction (been a long time since organic chem, mind you)= is a condensation rather than an addition (e.g. polyethylene ). Some water is exchanged at the business end of the anhydride during these reactions, bu= t there shouldn=D5t be much generated overall. (Just how long is beginning = to show, I think). Anyway, a low viscosity resin like VCD (Spurr=D5s resin) might = be the ticket. If you limit the amount of flexibilizer (DER 736, Quetol, etc.) = you can get these resins quite hard and the resulting sections, especially if= given a light carbon coating are relatively beam-stable.
Other things will cross-link epoxies, polyamines, for example but I=D5m y= et less aware of their polymerization reaction than for what the biologists around here use. The other commonly used class of resins are = the acrylics. Most of the ones in common use are proprietary formulations. = The old, conventional acrylics aren=D5t very beam stable.
Having embedded it, do you then plan ion milling? I=D5ve not done this w= ith an epoxy embedded powder sample but, epoxy glue lines in sandwitched (mostly= Si) tend to erode quickly, at my hands. Might be grim for a powder. If you t= ry the ultramicrotome, you=D5ll need something other than water to float your films on or do it dry (sounds li= ke you=D5d need a really hard resin for that to work at RT; we can=D5t do an= hydrous cryo sections in our lab, here in the cellar ;-). You might be able to g= et thin enough or nearly thin enough with a tripod polisher, but you=D5ll ne= ed to work out an anhydrous system for, at least, the final colloidal silica st= ep.
I think I=D5d try to work out a tripod system for this, $0.02. Good luck.
Cheers, John Heckman TEM Supervisor Center for Electron Optics Michigan State University
A number of people have asked for more details as to my previous query.
First some brief background. Cabot has developed a new reinforcing filler for elastomers, Carbon-Silica Dual Phase Fillers. Naturally we are working on characterizing several aspects of these new fillers.
I would like to assess the morphology of the silica phase by removing the carbon phase. Ashing the sample in bulk and viewing by TEM provides some useful information. I would like a higher level of information regarding the silica phase.
So what I have tried to do is image the dual-phase aggregates on a TEM substrate, then place the TEM substrate into a muffle furnace for 2 hours at 550C in air. The intention is to then view the ashed aggregates (now only silica). Unfortunately the substrates I have tried (germanium and gold) have decomposed. By decompose I mean the substrate is no longer on the grid or severely folded onto the grid bars. I am very surprised that this also happens with a gold substrate on a gold 400 mesh grid!!
The carbon in the dual phase filler begins to decompose about 500C. The dimension of the aggregates are in the range of 100nm give or take.
Regards,
Larry Murphy Group Leader, Analytical Section Cabot Corporation
I'm looking for the source of the quote: "There are lies, damn lies, and statistics."
No, this has nothing to do with interpretation of EDS spectra despite what the cynics may say!
Thanks in advance
Harry ------------------------------------------------ Opinions or statements expressed herein, rational or otherwise, do not necessarily reflect those of my employer.
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 Phone: (508) 750-1717 E-mail: crossman-at-osi.sylvania.com
Our web sites: www.sylvania.com www.siemens.com -- "Crossman, Harold" {crossman-at-osi.SYLVANIA.com}
} I have a question regarding what polymer resin to use for embedding a } sample for (TEM) observation. The sample is a powder, which is likely to } be moisture and CO2 sensitive. I am thus concerned that a condensation } polymer would induce changes in the sample. We have been grinding the } sample to produce a suspension for TEM specimen preparation. However, we } would like to use ion milling, as this will be more likely to preserve } the microstructure. } } Does anyone know of a good embedding polymer for a moisture and CO2 } sensitive sample, which is also high-vacuum compatible? } Is it possible to examine the powder without embedding? I have looked at sediment particles which have been suspended in water and placed on a grid. These have been examined after the water has evaporated. Since water is not good for your particles, another liquid would have to be used, but the procedure should work equally well. It is also very quick to try. Yours, Bill Tivol
We are using our new Ted Pella 3450 microwave for processing potato tissue and have some questions: 1) Do the fumes created by polymerization of LR White stay in the equipment and so might hinder future use for immunolabeling? 2) What would be the best grids for Protein A-gold labeling (on LR White)? 3) What is the best temp. limit for alcohol dehydrations? Thanking you in advance for your help... Bonnie Compas
I am posting the following query for a client of mine. Please respond to MICROSCOPY as usual, or to me privately off-line. Thanks.
Gib Ahlstrand
----------------------------------------------------------------------------- I have some encapsulated flavor particles primarily coated with modified starch and sucrose and coated secondarily with fat. To look at how the flavor particles are coated with fat, confocal microscopy is used. I have stained fat with nile blue. I wonder what is a good stain for modified starch. Any other staining tips you can give for fat & starch using confocal microscopy will be appreciated.
I have been having particular difficulty with a batch of fish eggs I have tried to prepare for SEM. They were fixed in glut. followed by osmium and then dehydrated to 100% EtOH. I have then dried them from liquid CO2 in a CPD. A few eggs have survived the drying intact, the rest either exploding or collapsing.
I have extended the periods in EtOH, and soaking in the CO2 (with lots of flushing), and have slowly vented the gas to minimize any 'shock'. I have tried taking the eggs from 100% EtOH through a graded series of Freon before drying, but the eggs shrivel up by the time I get to 50% Freon. I also tried drying from Peldri, going in a graded series from 100% EtOH to 100% Peldri and letting them soak in the liquid Peldri overnight. By that stage, they had again collapsed.
I guess freeze-drying might be my next move, but thought I'd ask if others may have suggestions on how to get these specimens through the CPD stage. I have processed fish eggs before, but these seem to be quite robustly encapsulated.
Thank you.
Carolyn J. Emerson email: cemerson-at-plato.ucs.mun.ca
Biology Department Memorial University St. John's, NF A1B 3X9 Tel: (709) 737-7515 Fax: (709) 737-3018
I recall it was Mark Twain in "Life on the Mississippi". It was connected to a discussion as to how fast the mouth of the Mississippi was moving north and how soon it would be in Tennessee instead of Louisiana.
At 09:49 AM 10/29/97 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
According to my source, The New Dictionary of Thoughts by T. Edwards et al and published in 1959 (okay, I ain't that new either) by Standard Book Company, the quote is actually:
There are three kinds of lies - lies, damnable lies, and statistics. Commander Holloway H. Frost
I pesonally prefer:
Statistics are no substitue for judgement. Henry Clay
Shalom from Jerusalem, Azriel Gorski
On Wed, 29 Oct 1997, Crossman, Harold wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I'm looking for the source of the quote: "There are lies, damn lies, } and statistics." } } No, this has nothing to do with interpretation of EDS spectra despite } what the cynics may say! } } Thanks in advance } } Harry } ------------------------------------------------ } Opinions or statements expressed herein, rational or otherwise, do not } necessarily reflect those of my employer. } } Harold J. Crossman } OSRAM SYLVANIA INC. } Lighting Research Center } 71 Cherry Hill Dr. } Beverly, MA 01915 } Phone: (508) 750-1717 } E-mail: crossman-at-osi.sylvania.com } } Our web sites: www.sylvania.com } www.siemens.com } -- } "Crossman, Harold" {crossman-at-osi.SYLVANIA.com} }
I get to much mail. WHY do people not reply directly to inquiry posters'!
______________________________________________________ Anand Patel Harvard School of Public Health 94 Beacon Street #78 Cardiovascular Biology Laboratory Somerville, MA 02143 Building II, Room 127 677 Huntington Avenue tel & fax: (617) 354-5132 Boston, MA 02115 USA
} Hello, Does anyone know how to open an IP Lab 12 bit greyscale image file } captured from a Photometrics Sensys CCD camera, as a Raw file in } Photoshop? All you have to do is to convert the 12 bit image to 8 bit. You can do this with IPLab, go to Math and select "to byte as shown" and save your file. Now you'll be able to open your stuff in Photoshop. Let me know if you have any problem
--Ciprian Have fun and keep the sun on your back and a smile on your face. __________________________________________________________ Ciprian A. Almonte University of Pittsburgh Center for Biologic Imaging Pittsburgh, PA 15261
} } } Hello, Does anyone know how to open an IP Lab 12 bit greyscale image file } captured from a Photometrics Sensys CCD camera, as a Raw file in } Photoshop? } } Bob } Morphology core Lab } U of Washington } Hi Bob, It seems that IPLab must be gaining popularity, as this question has been posed a few times recently. The solution, according to Micheal Mort, Ph.D. (president of Scanalytics, Inc., the makers of IPLab software) is to go to the Math menu (once the image is contrast adjusted), and select the command "to byte as shown" then save as a tiff file. Dr. Mort has been most gracious and prompt with my questions regarding their software, and can be reached via email at mmort-at-iplab.com He also informed me that they have software IPLab/Windows at a discount price (since you already proably have IPLabs/Mac). I have no financial interest in any of the above mention firms, just someone like yourself, learning as I go......
Randy Nessler rnessler-at-emiris.iaf.uiowa.edu Views expressed are my own.
Hi Carolyn, You might try some of the mordant techniques using glutaraldehyde, tannic acid, guanidine HCl and osmium by: Gamliel, Scanning Elec. Microsc. 1985: IV; pp1649-1662, 1985. or Osmium, tannic acid, uranyl acetate as per: Shroeter, etal., J. Elect. Microsc. Techn. v1 pp219-225, 1984.
Judy Murphy published two nice reviews covering non-coating techniques which may also help to strengthen cells against collapse: Scanning Electron Microsc. 1978, vol II, pp 175-194 Scanning Electron Microsc. 1980 , vol I, pp 209-
Klaus Peters also published a paper titled "Improved handling of structural fragile cell-biological specimens by the exchange method." J. of Microscopy v 118, pp 429-441.
I could dig them out and FAX to you if you do not have access to these journals.
good luck
cheers
Edward J. Basgall, PhD The Pennsylvania State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618 http://www.personal.psu.edu/ejb11/ Privilege does not absolve one of ecological responsibility.
I am trying to do the same thing, but on yougert. I am using fast green for protein and nile blue for fat, but what fluorochrome can be used in the 647nm wavelength of the Argon/Krypton laser to identify starch?
list
William R. McManus Electron Microscopy Facility Department of Biology Utah State University Logan UT 84322-5305 1-801-797-1920 billEMac-at-cc.usu.edu
On Wed, 29 Oct 1997 09:45:32 -0800 (PST) BECOMPAS-at-CSUPomona.edu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } We are using our new Ted Pella 3450 microwave for processing potato tissue } and have some questions:
Bonnie: you will probably get lots of different responses to these questions. Here are mine, based on at least some experience with the same equipment:
} 1) Do the fumes created by polymerization of LR White stay in the equipment } and so might hinder future use for immunolabeling?
Can you vent your 3450 to the outside? Certainly, this would be recommended. We have noticed that if samples of LR White are not covered during polymerization, the media does seem to sublimate, then recondenses and polymerizes on all surfaces (this is our experience during the polymerization of LR White in a nitrogen rich chamber heated to 60C). Using the microwave, you should consider submersing beem capsules, sealed with parafilm under the cap, in water. This will insure that LR white fumes do not enter the microwave during polymerization. Use the temperture limiting probe to limit the temperture of the water to 70 C and polymerize for 60 minutes, or set to 80C and polymerize for 45 minutes. A 500 ml beaker with recirculated water at 10 to 20 C should also be included in the microwave during polymerization.
} 2) What would be the best grids for Protein A-gold labeling (on LR White)?
We use formvar coated Nickel grids. We prefer slot grids, but use whatever you prefer. You may want to use uncoated nickel grids if you wish to label both sides for a double labeling protocol.
} 3) What is the best temp. limit for alcohol dehydrations?
A progressively lower temperture scheme is best. Start with 30% EtOH at 0C for 10 min, then lower to -10 C for another 10 min. Add 50% at -10C, then lower to -20C. All subsequent steps to 1:3 90% EtOH:LRW should be at -20C. We also leave the samples overnight in 100 % LRW at -20 C, then raise the temp to ambient for another hour or so prior to thermal polymerization. We might be to careful. You could work in the cold room (brrrrr!) to obtain -20C.
Feel free to phone if you would like to trade secrets. (503)- 221-3434.
} Thanking you in advance for your help... } Bonnie Compas
In regard to Lawrence Murphy's request, the utilization of a low energy plasma containing oxygen would work to selectively remove the carbonaceous material without effecting the silica. In this process, disassociated oxygen is created by the plasma. The oxygen radicals combine chemically with the carbon, converting it to CO and CO2. By utilizing a plasma type which possesses ion energies of less than 25 eV, the carbon is reduced without any alteration to the substrate.
Please feel free to contact me directly for more specific information relating to the process.
Kind regards,
Paul
Paul E. Fischione, President E.A. Fischione Instruments, Inc. 9003 Corporate Circle Export, PA 15632 Phone 412-325-5444 FAX 412-325-5443 e-mail paul.fischione-at-internetmci.com www.fischione.com ---------- } From: Lawrence_Murphy-at-cabot-corp.com } To: microscopy-at-Sparc5.Microscopy.Com } Subject: Re: Ashing Samples on TEM Substrates } Date: Wednesday, October 29, 1997 10:24 AM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } A number of people have asked for more details as to my previous query. } } } First some brief background. Cabot has developed a new reinforcing filler } for elastomers, Carbon-Silica Dual Phase Fillers. Naturally we are working } on characterizing several aspects of these new fillers. } } I would like to assess the morphology of the silica phase by removing the } carbon phase. Ashing the sample in bulk and viewing by TEM provides some } useful information. I would like a higher level of information regarding } the silica phase. } } So what I have tried to do is image the dual-phase aggregates on a TEM } substrate, then place the TEM substrate into a muffle furnace for 2 hours } at 550C in air. The intention is to then view the ashed aggregates (now } only silica). Unfortunately the substrates I have tried (germanium and } gold) have decomposed. By decompose I mean the substrate is no longer on } the grid or severely folded onto the grid bars. I am very surprised that } this also happens with a gold substrate on a gold 400 mesh grid!! } } The carbon in the dual phase filler begins to decompose about 500C. The } dimension of the aggregates are in the range of 100nm give or take. } } Regards, } } Larry Murphy } Group Leader, Analytical Section } Cabot Corporation } }
Is your web site the best kept secret on the Internet?
We'll promote it to 50 search engines and indexes for $85 and complete the job in 2 business days. Satisfaction is guaranteed!
If you have a great product, but are not getting many inquiries from your Web site, you may not be adequately listed on the Web's search engines and indexes.
Millions of viewers daily use these facilities to find the products and services they are looking for. But if your site is not listed, no one will see it.
Listings on most of these services are free. However, locating and filling out the forms required to get a listing can take several days, and most people just don't have the time to do it.
That is why we offer a web site promotion service.
WHAT'S THE DEAL?
We will submit your site to 50 indexes and search engines for $85. We will accept the return of this E-mail, with the form below filled out, as an order. We will bill you upon completion of the promotion. Our terms are net 15 days from date of invoice. Satisfaction guaranteed!
HOW LONG WILL IT TAKE?
Generally, we complete the submissions within 48 hours of receiving your order. It can take any individual search engine or index up to three weeks to process your submission, although most are much faster.
WHAT SEARCH ENGINES AND INDEXES ARE INCLUDED IN THE PROMOTION?
The list changes from time to time. This is our current list:
Alta Vista, BC Internet, BizCardz Business Directory, BizWeb, Excite, Galaxy, HotBot, Infoseek, InfoSpace, Jayde Online Directory, JumpCity JumpLink, Linkcentre Directory, LinkMonster, Lycos, Magellan, Manufacturers Information Network, Net Happenings, Net Mall, Net-Announce, New Page List, New Riders WWW Yellow Pages, Northern Light, One World Plaza, Open Text Web Index, PageHost A-Z, PeekABoo, Project Cool, Scrub The Web, Seven Wonders, Sserv, Starting Point, The Galactic Galaxy, The Weekly Bookmark, True North,TurnPike, Unlock:The Information Exchange, Web 100, Web Crawler, Web Walker, Web World Internet Directory, WebVenture Hotlist, What's New, WhatUSeek, Where2Go, World Wide Business Yellow Pages, Wow! Web Wonders!, WWW Worm, YelloWWWeb, Your WebScout
HOW WILL I KNOW THAT YOU HAVE PROMOTED MY SITE?
When we have completed the promotion, we will send you an HTML file as an attachment to your E-mail bill. Save this file to your disk, and view it through your Web browser. It provides links to the search engine we submitted your site to, plus any comments we received from them when we did it.
ARE THERE ANY GUARANTEES?
We do not require prepayment. Your satisfaction is guaranteed or you don't pay the bill.
WHO IS OWL'S EYE PRODUCTIONS?
We are a web site promotion company located at:
Owl's Eye Productions, Inc. 260 E. Main Street Brewster, NY 10509 Phone: (914) 278-4933 Fax: (914) 278-4507 Email: owl-at-secretwebsite.com
HOW DO I ORDER?
The easiest way to order is by e-mail. Just hit the REPLY button on your e-mail program and fill out the following information. (This information will be posted to the search engines/indexes):
Your name: Company Name: Address: City: State/Prov: Zip/Postal Code: Telephone: Fax: Email address: URL: http:// Site Title: Description (about 25 words): Key words (maximum of 25, in descending order of importance):
Proofs (Where shall we e-mail proofs):
If billing a different address, please complete the following:
Terms: By returning this document via Email, you agree as follows: You have the authority to purchase this service on behalf of your company. Terms are net 15 days. Accounts sent to collections will be liable for collection costs. You agree to protect and indemnify Owl's Eye Productions, Inc. in any claim for libel, copyright violations, plagiarism, or privacy and other suits or claims based on the content or subject matter of your site.
WHAT HAPPENS NEXT?
When we receive your order, we will input the information into our system, and send you a proof.
After we process any corrections, we will run your promotion, capturing any comments from search engines as we go. We will incorporate these into an HTML-formatted report to you, which we will attach to your bill. ===Web Promotions=====Press Releases=====Link Exchanges========= Owl's Eye Productions, Inc. 260 E. Main Street Brewster, NY 10509 Ph: 914-278-4933 Fx: 914-278-4507 E-mail: owlseye-at-secretwebsite.com
I have troubles with a box of 10 year old Philips W SEM filaments. It is very difficult or almost impossible to obtain a good saturation or worse, increasing the filament current the beam current decrease. With new ones no problem. These filaments were forgotten for some year but were always stored in a clean place. My question is: is this an aging effect or the filaments are defective? Is there any possibilty of recovery ? (for instance cleaning with an acid bath ?) Thanks for help
------------------------------------------- Giorgio Gasparotto Dipartimento di Scienze della Terra e Geo-Ambientali Piazza di Porta S. Donato 1 40126 Bologna Italy Tel. 51 243.556 FAX 51 243.336 WWW: http://geode.geomin.unibo.it Internet e-mail gaspar-at-geomin.unibo.it -------------------------------------------
Anand wrote: } } I get to much mail. WHY do people not reply directly to inquiry posters'! } Very simple. Other people might be following the thread and waiting for the replies. Anything that might be of general interest should be posted to the whole list. It saves us sending inquiries about possible replies to the inquiry.
That is my opinion, anyway.
-- Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 86 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www_scem.csb.ki.se/pages/philip.html
} We are using our new Ted Pella 3450 microwave for processing potato tissue } and have some questions: } 1) Do the fumes created by polymerization of LR White stay in the equipment } and so might hinder future use for immunolabeling?
This is a function of the instrument's ventilation system. If the vent fan is sufficiently powerful (the one in our microwaves is rated at 100 cubic feet per minute), there should be no problem with fumes.
{snip}
} 3) What is the best temp. limit for alcohol dehydrations?
In my experience, the alcohols should be kept at or below 40 degrees C.
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
Thanks for the rapid and, as always, very informative responses to my question about collapsing/exploding fish eggs as they are dried for SEM viewing. Among the suggestions were:
1. Use OTOTO methods or other non-coating techniques to 'strengthen' the sample and reduce the possibility of shrinkage.
2. Puncture or cut the eggs in half to allow passage of fluids (not really feasible in my case, but no doubt useful for others).
3. Try a cryo-stage examination.
4. Try processing/drying from HMDS.
5. Do not allow ANY exposure to air during the handling of the eggs.
6. Keep the sample from being directlly exposed to the incoming CO2, ie invert the coverships in the dryer.
OTOTO and HMDS seem the next routes for me to go. THanks again to those who responded.
Carolyn J. Emerson email: cemerson-at-plato.ucs.mun.ca
Biology Department Memorial University St. John's, NF A1B 3X9 Tel: (709) 737-7515 Fax: (709) 737-3018
Thanks to everyone who replied. I'm going to conduct a seance to see if I can get Benjamin Disraeli, Mark Twain, William Shakespeare, the authors of the Bible, Commander Holloway H. Frost, and Winston Churchill in the same venue to argue the TRUE source. If possible, the debate will be moderated by Peter J. Stratham of Oxford Instruments.
------------------------------------------------ Opinions or statements expressed herein, rational or otherwise, do not necessarily reflect those of my employer.
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 Phone: (508) 750-1717 E-mail: crossman-at-osi.sylvania.com
Our web sites: www.sylvania.com www.siemens.com --
Help! My nephew was given a E.Leitz-Wetzlar microscope and wants to trade it for a piece of art. He has no idea what it is worth and will probably come out on the short end of stick. Here is the info he has given me - model# 264460, dated July 1926. The microscope is brass and black ceramic, has 2 lens for top, testing oil emergence? and comes with black case that says, "The Max Worcher & Sons Co. Surgical Instruments, Cinn. Ohio. If anyone can help I would be grateful as I would hate to see him make a mistake. Thank You.
"Very simple. Other people might be following the thread and waiting for the replies. Anything that might be of general interest should be posted to the whole list. It saves us sending inquiries about possible replies to the inquiry."
I agree - I often read about problems or solutions I didn't know existed, but which are nonetheless relevant to my work. A listserver whose sole purpose is to initiate a series of private conversations seems like a waste of technology to me.
Marie
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-131 University of Connecticut Storrs, CT 06269 Ph: 860-486-3588 Fax: 860-486-1936
The 24V power supply on my AMRAY 1000B SEM has recently blown out. Are there any kind souls out there with a spare power supply or know of a source where this one can be rebuilt?
Many thanks,
Dennis Shubitowski University of Michigan School of Dentistry dshubito-at-umich.edu
Developmental Biologist Department of Zoology Miami University Oxford, Ohio
We are seeking applicants for an Assistant Professor tenure-track position to begin in August, 1998. Ph.D. in zoology or biology and postdoctoral experience required. Individuals with expertise in any area of Animal Developmental Biology are invited to apply, but preference will be given to those who use electron microscopy as a major research tool. We expect this person to develop an independent research program in developmental biology that will enhance the department's research capabilities.
Teaching responsibilities include: (1) a sophomore level course in developmental biology; (2) participation in a team-taught introductory biology course; and (3) and advanced course in a specialty area. The successful applicant will be expected to seek external funding to support his/her research and to supervise and advise graduate and undergraduate students. Advancement will be based on teaching, research, and professional service, with primary emphasis on teaching and research.
Miami University is a state-assisted institution in SW Ohio. The department has excellent research facilities; the EM facility is well-equipped for SEM, TEM, cryopreservation, and confocal microscopy (see http://www.muohio.edu/~zoocswis for more details about the department and our facilities). The department has strong Ph.D. and M.S. programs, 32 faculty members, several postdoctoral researchers, and 50 graduate students on the Oxford campus.
Interested persons should submit a curriculum vitae, a statement of teaching philosophy, a description of current research and long-term research interests, and should arrange for three letters of recommendation and transcripts of graduate and undergraduate academic work to be sent to:
Dr. Douglas H. Taylor, Chair of Zoology, Miami University, Oxford, OH 45056.
Review of applications will begin on 1 December, 1997, and continue until the position is filled. Miami University offers equal opportunity in employment and education.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"640K ought to be enough for anybody." -- Bill Gates, 1981
} Dear List, } } I am trying to do the same thing, but on yougert. I am using fast } green } for protein and nile blue for fat, but what fluorochrome can be used } in the 647nm wavelength of the Argon/Krypton laser to identify } starch? } } billEMac-at-cc.usu.edu
I'm not a confocalist (watch carefully as I insert my foot in my mouth) but starch will stain with PAS, and Schiff's is fluorescent... acriflavine-Schiff's (excitation 480, emission 550-600) will fluoresce yellowish orange, pararosaniline-Schiff's (excitation 560, emission 625) will glow red.
I realise that neither will work at 647nm, but do you have any other wavelengths available?
Regards
Stephen Edgar
Electron Microscope Unit, Pathology Department School of Medicine University of Auckland Private Bag 92019 Auckland New Zealand
Medical research laboratory has an opening for a full time position as Research Technician. Candidates should have a Bachelor's Degree and previous experience in many of the following techniques: tissue processing, microtomy, light and electron microscopy, photography and darkroom, immunohistochemistry, and in situ hybridization. Familiarity with computers is useful but not essential.
- Available Immediately -
Please send a resume and a list of 3 personal references (address & phone number) to: David H. Hall, Ph.D. Department of Neuroscience Albert Einstein College of Medicine 1410 Pelham Parkway Bronx, NY 10461
Send by mail [or by email to: hall-at-aecom.yu.edu]
AECOM is located in a residential section of the north Bronx, with good subway, bus and highway connections to Manhattan, Long Island, Westchester County, and New Jersey.
Anand wrote 10/30/97: } =20 } I get to much mail. WHY do people not reply directly to inquiry = posters'! }
Philip Koeck wrote 10/30/97: "Very simple. Other people might be following the thread and=20 waiting for the replies. Anything that might be of general interest should be posted to the whole list. It saves us sending inquiries about possible replies to the inquiry.
That is my opinion, anyway."
--=20 Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 86 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www_scem.csb.ki.se/pages/philip.html
The fact that a lot of postings/messages is sent via the MSA-Server is = obvious. Sometimes I also think about how to manage that mass of = informations. BUT: still I am reading them and if of interest I store = them according to a list of items created from Microsoft Exchange. The only problem I get with messages containing no "header" or = "concern", sometimes it is a RE, but it is sent like a "Q: question" so = I loose time in storing such informations for rewriting the = "concerns"-header. So my request to Users of the MSA-Server would be: Please adhere to the = regulations of the Server. Thank you very much.
Dr. Wolfgang MUSS Department of Pathology, LKA EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
Dear all, all silicone rubber mold "packages" as requested are mailed since = Tuesday, 28th of October, including product data sheets, instructions as = well as at least one self fabricated mold. 30 colleagues were interested in getting such information.
Within the last 3 weeks I fabricated approx. 45 molds which had to stand = 2 days for aeration on air (condensation process), then they were = tempered for at least 30 hours at 140 degr.C. Those embedding molds are tested and approved for use with epoxide-resin = (EPON 812 substitute Glycid-ether 100 from SERVA/BIOPRODUCTS) but not = with hydrophilic resins like LRWhite or Lowicryls.
I hope that you are satisfied with the informations you got.=20
For all those who were included in the first "mail" (~ 10/7-10/97): for U.S.A.residents: please call or contact the=20 WACKER SILICONE COMPANY at ADRIAN, Michigan (as indicated in my = text-info).
For residents in other countries than U.S.A.:
As I was informed by WACKER SILICONE GmbH Munich, all inquiries = concerning the ELASTOSIL R silicone masses, orders, info-requests etc. = should be addressed to:
DRAWIN-Vertriebs-GmbH c/o Mr. Helmut Klug or Mrs. Cornelia POHL=20 (they know about my information to you) Rudolf-Diesel-Strasse 15 D-85 521 OTTOBRUNN/RIEMERLING (which is situated near MUNICH) phone: ++49++ 89 - 6 08 69 - 0 FAX: ++49++ 89 - 6 08 69 - 250
It is a sub of WACKER SILICONES and unfortunately they (WACKER) didn=B4t = mention this address in their info-brochures.
If there is any questions more, please e-mail directly. If there is anybody of the colleagues who told me his interest but = didn=B4t get the package within the next week or so, please also e-mail = directly.=20
If there is anybody else who wishes to receive also that information = package, please request it also preferably by direct e-mail.=20
At the moment I=B4m out of glass-flasks/bottles for evacuating the = silicone rubber mass. Also I have to order new silicone mass, since all = of my own has gone. So please be patient.
I greatly should appreciate a short note when/that you got your = "package".
I thank you all for your interest and wish "good luck" for the first = self-fabricated mold meeting your special needs.
Best regards and have a nice day/night/weekend
Dr. Wolfgang MUSS Department of Pathology, LKA EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
} } } "Only the small minded will keep things in order the genius will overlook the chaos". { { { Today I am small minded.
Anand wrote 10/30/97: } =20 } I get to much mail. WHY do people not reply directly to inquiry = posters'! }
Philip Koeck wrote 10/30/97: "Very simple. Other people might be following the thread and=20 waiting for the replies. Anything that might be of general interest should be posted to the whole list. It saves us sending inquiries about possible replies to the inquiry.
That is my opinion, anyway."
--=20 Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 86 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www_scem.csb.ki.se/pages/philip.html
The fact that a lot of postings/messages is sent via the MSA-Server is = obvious. Sometimes I also think about how to manage that mass of = informations. BUT: still I am reading them and if of interest I store = them according to a list of items created from Microsoft Exchange. The only problem I get with messages containing no "header" or = "concern", sometimes it is a RE, but it is sent like a "Q: question" so = I loose time in storing such informations for rewriting the = "concerns"-header. So my request to Users of the MSA-Server would be: Please adhere to the = regulations of the Server. Thank you very much.
Dr. Wolfgang MUSS Department of Pathology, LKA EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
To AParent and microscopist: In the end that nephew will do has he pleases. There are some considerations which I have made: Art, presumably a painting, can be very appealing for some time, but after some years they seem dated or inappropriate. The truest general saying about art is "today's art is tomorrows trash" - and the exceptions (I venture less than 1%) prove that rule. I see art as decorative materials and not an investment.
That microscope, when properly displayed is an attractive statue. In twenty years time, chances are overwhelming that the microscope will have doubled in price and that the other piece of art will be very hard to sell at any price. Incidentally, in the history section of our links pages is an internal links to my two antique Leitz microscopes. I believe that either should be worth about US$2000 in the right market - sorry they are not for sale; they no longer make antique microscopes, just plenty of paintings. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au
---------- } From: AParent103-at-aol.com } } Help! My nephew was given a E.Leitz-Wetzlar microscope and wants to trade it } for a piece of art. He has no idea what it is worth and will probably come } out on the short end of stick. Here is the info he has given me - model# } 264460, dated July 1926. The microscope is brass and black ceramic, has 2 } lens for top, testing oil emergence? and comes with black case that says, } "The Max Worcher & Sons Co. Surgical Instruments, Cinn. Ohio. If anyone can } help I would be grateful as I would hate to see him make a mistake. Thank } You.
I spend most of each day preparing and analyzing samples using light microscopy and FT-IR microscopy. Fingerprints are more than a nuisance, so its frequent trips to the sink for a little soap and water. In the winter I find my fingertips become dry and cracked. Lotion is out during work hours. Cotton gloves shed linters. Finger cots provide some relief.
Anybody have other suggestions?
James "Fingers" Martin :) Williamstown Art Conservation Center
Stephen Edgar wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } } Dear List, } } } } I am trying to do the same thing, but on yougert. I am using fast } } green } } for protein and nile blue for fat, but what fluorochrome can be used } } in the 647nm wavelength of the Argon/Krypton laser to identify } } starch? } } } } billEMac-at-cc.usu.edu } } I'm not a confocalist (watch carefully as I insert my foot in my } mouth) but starch will stain with PAS, and Schiff's is fluorescent... } acriflavine-Schiff's (excitation 480, emission 550-600) will } fluoresce yellowish orange, pararosaniline-Schiff's (excitation 560, } emission 625) will glow red. } } I realise that neither will work at 647nm, but do you have any other } wavelengths available? } } Regards } } Stephen Edgar } } Electron Microscope Unit, Pathology Department } School of Medicine } University of Auckland } Private Bag 92019 } Auckland } New Zealand } } email address: s.edgar-at-auckland.ac.nz } Phone : +64-9-3737599 extn 6473 (GMT + 12h) } Fax : +64-9-3737459
Gentlemen,
An interesting alternative to trying to find a third fluorophore: Starch has a unique response to polarized light: it exhibits a maltese cross. Just as you might combine regular epi-fluorescence with, let's say, transmitted light phase contrast, why couldn't you combine fluorescence with polarized light? It would really only take adding one polarizer over the transmitted light source and a second, in crossed position, to the barrier filter ... or, if you have access internally to the confocal system, somewhere before the detector.
If you try this combination, please send me an image.
Best regards,
Barbara Foster President Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108 PH: (413)746-6931 FX: (413)746-9311 email:mme-at-map.com --------------------------------------------------------------------------------------------------------------------------------- ********** Microscopy/Microscopy Education ********** America’s First National Consortium of Microscopy Experts Specializing in Customized, On-site Training in all areas of Microscopy, Sample Prep, and Image Analysis
I've got a question about plasma "etching" or "ashing" procedure to remove outer organic surfaces of polymer samples.
Another lab who did this for us in the past had a procedure where they did the etching in two steps: 1. oxygen plasma 2. argon plasma. (The other lab won't talk to us anymore, but that's another story...)
I can't figure out what what the argon plasma is supposed to do. The function of the oxygen plasma is clear: to oxidize the carbon and hydrogen, producing volatile substances. A mixed oxygen/argon plasma might make sense too, to control the ablation rate for instance. But the reason for doing pure oxygen and then pure argon eludes me.
Am I being dense? Can anybody give me a tip on this? (I'm a chemist, for goodness sake, but I still can't figure it out!)
We have found that the `pseudo-leather' gloves available from Agar Scientific (and I guess most other em accessory suppliers) are good for long term wear. The surface eventually starts to break up with wear but until then they are fine. They are the only gloves we will wear when handling HT components as all others we have tried leave something behind on the surface. They retail here at about 3.2 pounds sterling a pair.
Ron
Usual disclaimer - I am purely a satisfied customer. =========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
} } after several unsuccessful attempts to stain PE ultrathin cuts with the } Kanig method I would like to ask if somebody has been successful and is } willing to share his protocol with me. } The specimen is already cut with an cryo-ultramicrotome and the cuts are } lying on copper grids. }
Here at Reading, many years ago, we used to do chlorosulphonation of PE by (more or less) the Kanig method. One always stained FIRST, then cut the section.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
I am looking for documentation useful to my generating an operating procedure in a Good Laboratory Practice (GLP) environment for documenting transmission, archival, and handling of telepathology case sessions. Could anyone direct me to an area where I might review other protocols currenlty acceptable and in use Greg Argentieri Novartis Pharmaceuticals Corp 59 Rt 10 Bldg 406 Rm 121 East Hanover, NJ 07936 973-503-8617 Fax 973-503-6339 E-mail Gregory.Argentieri-at-pharm.novartis.com
One can get fairly small refrigerators that can be dedicated to osmium and glut. We double containerize the os solutions. First in a Teflon screw top bottle and then in a jar with a screw top and seal in the lid (intended to food preservation) We hope this minimizes the os vapor leackage. We store cacodylate in the regular chemical refrigerator. We also have a refrigerator reserved for biologicals such as enzymes and antibodies, away from any fixative vapors that might denature them. Os waste is stored in large screw top bottles which originally contained solvents. It is in the fume hood at room temp until it is collected by our safety dept. } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } . At 11:03 AM 10/31/97 +1100, you wrote:
} Fellows Biological Users } } I am seeking information on the type of refrigerator and types of storage } currently used by biological departments using: } } osmium tetroxide + waste, and } } glutaradehyde } } sodium cacodylate } } } Thankyou in advance for your help } } } Barry } EM Unit } UNSW } } } ******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab PO Box 118525 Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
You need to say if you are handling any chemicals that require protection, in which case you might try nitrile gloves, which are thin, strong, chemically resistant, and do not cause latex allergies. If you don't need gloves, you could try something moisturizing after washing. I use a product called Eucerin, a water-in-oil emulsion that's expensive, but a jar will last you a long time.
Leonard Corwin, Principal Research Chemist Fort Dodge Animal Health, Animal Health Research Analytical Princeton, NJ 08543-0400 corwinl-at-pt.cyanamid.com
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Here's a new thread. Hand care for microscopists.
I spend most of each day preparing and analyzing samples using light microscopy and FT-IR microscopy. Fingerprints are more than a nuisance, so its frequent trips to the sink for a little soap and water. In the winter I find my fingertips become dry and cracked. Lotion is out during work hours. Cotton gloves shed linters. Finger cots provide some relief.
Anybody have other suggestions?
James "Fingers" Martin :) Williamstown Art Conservation Center
} Here's a new thread. Hand care for microscopists. } } I spend most of each day preparing and analyzing samples using light } microscopy and FT-IR microscopy. Fingerprints are more than a nuisance, } so its frequent trips to the sink for a little soap and water. In the } winter I find my fingertips become dry and cracked. Lotion is out during } work hours. Cotton gloves shed linters. Finger cots provide some relief. } } Anybody have other suggestions? } } James "Fingers" Martin :) } Williamstown Art Conservation Center
I use Vaseline Intensive Care lotion, or any similar hand lotion. Small amount 2-3 times a day work better than a larger amount once a day.
After polishing my sample I immersed it in acetone to dissolve the cement and after it had separated from the glass support I examined it by OM . At that point I noticed that a thin solid film of the colloidal, non-crystallizing silica, had formed on the surface of the sample. Is there a safe way of removing this film ?? I've tried soaking the sample in the colloidal suspension and in water but it did no seem to help.
I suspect the film formed because I failed to wash the sample properly before putting it in the acetone but I hate the idea of starting all over again.
I need some help on taking picture using a Zeiss STEMI SR. Some how the image in the camera is not focused uniformly. The image (and therefor the picture) is best focused at the central spot, and becomes blur at the edges, most noticeably on one side. When I raise or lower the scope (or stage), the image kind of move laterally, not up and down. I could not figure out why, except suspecting the alignment of the scope may be off. Any suggestions and comments are welcome.
I use a normal refrigerator for storage of all chemicals. We are required to label all refrigerators at the facility. Those refrigerators for food storage are labeled "For Food Only". Those refrigerators which contain chemicals (any type of chemical) are labeled: "No Food or Beverage / BioHazard" or something to that effect. The labels are on the outside of the refrigerators and are fairly large and usually brightly colored.
Glutaraldehyde is stored in regular screw-cap bottles.
Osmium crystals are stored in the original shipping vials and packaging in the refrigerator until they are diluted for use.
Osmium crystals are solubilized accordingly: The whole vial containing crystals is dropped into water or buffer to solubilize. The solution is made up in a 25-30 ml glass-stoppered reagent bottle, this bottle is placed inside a second slightly larger glass screw-cap bottle. The second bottle contains cotton in the bottom to cushion the first bottle. The second + first bottles are then placed inside a third still larger, about 550ml capacity plastic Bel-Art Products jar. The Bel-Art jar is brown with a white, opaque screw cap which eliminates light. We also put cotton or paper towelling in the bottom of the Bel-Art jar to cushion the glass bottles. The outside of both glass bottles are wrapped with parafilm before placing in the next bottle. It's not necessary to wrap the outside of the Bel-Art jar with parafilm. The whole lot is left in the hood overnight to solubilize the osmium.
For storage: The series of nested bottles/jars is placed into a normal refrigerator.
Osmium is extremely volatile and you will find that the parafilm becomes black upon storage indicating leakage.
Following the above storage procedure, we've never had a problem with a blackened refrigerator but I've heard of that happening in other places.
At 11:03 AM 10/31/1997 +1100, Barry Searle wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Delilah F. Wood United States Department of Agriculture Western Regional Research Center 800 Buchanan Street Albany, CA 94710
We have ethe same problem since we handle clinical samples and are constantly washing with antiseptic soap.
Sleep in gloves with lots of Vaseline on your hands. In the day time, chapstick applied just to the edges of the cracks (not all over your hands) can help if cracks are not on the very tips. At least you can apply it at lunch, etc. You can glue cracks together with superglue (a hint from my surgeon friend). Eucerine is a good hand cream recommended by my pharmacist; you might also try bag balm (for cows utters, available at farm supply stores) for times when you aren't at work.
Sara Miller
On Thu, 30 Oct 1997, James Martin wrote:
} Date: Thu, 30 Oct 1997 20:40:20 -0500 (EST) } From: James Martin {James.S.Martin-at-williams.edu} } To: microscopy-at-sparc5.microscopy.com } Subject: hand care for microscopists } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Here's a new thread. Hand care for microscopists. } } I spend most of each day preparing and analyzing samples using light } microscopy and FT-IR microscopy. Fingerprints are more than a nuisance, } so its frequent trips to the sink for a little soap and water. In the } winter I find my fingertips become dry and cracked. Lotion is out during } work hours. Cotton gloves shed linters. Finger cots provide some relief. } } Anybody have other suggestions? } } James "Fingers" Martin :) } Williamstown Art Conservation Center }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Our GW Champerscope works great. Since we installed it, no one has hit = the polepiece yet with the ancillary detectors. We have had it attached = to the JEOL 35C and the Hitachi 520 and plan to put it on our Electroscan = depending on the geometry AND once it gets installed.
The standard disclaimer applies of no financial interest in the above.
Judy Murphy Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207 209/954-5284 __________________________________________________________________________= _____
Hi:
I am thinking of getting an IR viewing system for our SEM chamber. Any advice or true to life adventures you can share?
Thanks.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
------------------ RFC822 Header Follows ------------------ Received: by sjdccd.cc.ca.us with ADMIN;31 Oct 1997 08:54:03 -0800 Received: from Sparc5.Microscopy.Com (206.69.208.10) by = ms.sjdccd.cc.ca.us with SMTP (Eudora Internet Mail Server 1.2); Thu, 30 Oct 1997 18:53:06 = -0800 Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com = (8.6.11/8.6.11) id SAA23594 for dist-Microscopy; Thu, 30 Oct 1997 = 18:24:23 -0600 Received: from cats.ucsc.edu (rumpleteazer.UCSC.EDU [128.114.129.45]) by = Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id SAA23590 for = {microscopy-at-sparc5.microscopy.com} ; Thu, 30 Oct 1997 18:24:22 -0600 Received: from cats-po-1 (root-at-cats-po-1.UCSC.EDU [128.114.129.22]) by cats.ucsc.edu (8.8.5/8.8.4.cats-athena) with SMTP id QAA16740 for {microscopy-at-sparc5.microscopy.com} ; Thu, 30 Oct 1997 = 16:35:41 -0800 (PST) Received: from [128.114.25.225] by cats-po-1 (8.6.13/4.8) id QAA13257; = Thu, 30 Oct 1997 16:35:27 -0800 Message-Id: {199710310035.QAA13257-at-cats-po-1} Mime-Version: 1.0 Content-Type: text/plain; charset=3D"us-ascii"
{fontfamily} {param} New_Century_Schlbk {/param} TWO RESEARCH ASSOCIATE POSITIONS AVAILABLE
(1) Research Associate position in a project to determine the structure of crossbridges in contracting insect flight muscle. Fast freezing followed by freeze substitution enables us to trap force bearing myosin crossbridges with millisecond time resolution. Electron tomography is then used to produce 3-D images that preserve the variable structure of the crossbridges for subsequent analysis. Mechanical records on stiffness and tension are recorded up to the moment of freezing.=20 Parallel time resolved X-ray diffraction data of the diffrent contractile states is also available with which to compare the EM data with native muscle structure. Methods have been developed over the past year to (1) classify and group the variable crossbridge forms for subsequent averaging that improves the signal to noise ratio in the reconstructions and (2) adjust and refine the atomic structure of the myosin head in order to fit it into the in situ envelope from the 3-D reconstruction. The research offers a unique opportunity to learn electron tomography, correspondence analysis of 3-D motifs and atomic modeling within the envelope of a low resolution envelop and at the same time provide unique information on the structural changes that occur in situ in contracting muscle. The project involves primarily computing to obtain the 3-D images and computer modeling to interpret the structure.
Recent Publications:
Holger Schmitz, Mary C. Reedy, Michael K. Reedy, Richard T. Tregear, Hanspeter Winkler, Kenneth A. Taylor. Electron tomography of Insect =46light Muscle in Rigor and AMPPNP at 23=B0C. {italic} J. Mol. Biol. {/italic} 264, 279-301 (1996)
Holger Schmitz, Mary C. Reedy, Michael K. Reedy, Richard T. Tregear, Hanspeter Winkler, Kenneth A. Taylor. Tomographic 3-D Reconstruction of Insect Flight Muscle Partially Relaxed by AMPPNP and Ethylene Glycol. {italic} J. Cell Biol. {/italic} (in press, November, 1997)
(2) Research Associate position in a project to determine the structure of alpha-actinin by electron crystallography. Project is funded through January 2001. We have obtained crystals of several different isoforms of alpha-actinin using lipid monolayers. The crystals are large in extent and to date yeild structural information to at least 10 Angstroms resolution. =20
Applicants for both positions must have a PhD degree. Salary is negotiable based on relevent experience. =20
The successful applicant will become a member of the Structural Biology program at Florida State University that includes 3 protein X-ray crystallography groups, three macromolecular NMR groups, 2 EPR groups and 2 electron microscopy groups. Facilities for electron protein crystallography include the above mentioned CM300-FEG, a Philips CM120, 3 Gatan cryotransfer systems, a Gatan wide angle TV camera, a Perkin-Elmer PDS1010M densitometer, 2 surveying optical diffractometers. Inquiries and applications should be made to Dr. Kenneth Taylor, Institute of Molecular Biophysics, Florida State University, Tallahassee, FL 32306-4380. Tel: (850) 644-3357. FAX (850) 561-1406. E-mail: taylor-at-bio.fsu.edu. Applicants should provide a CV and the names and addresses of 3 former mentors or knowledgeable individuals who can provided reference letters. =20
Has anyone figured out how to segregate from other mail the e-mail originating in this forum?
I use Internet Mail & News (v4.70) bundled with Microsoft IE 3.02. The "Inbox Assistant" in "Mail" menu is meant to assist in automatic segregation to sub-folders, but I have been unable to make it work with stuff from the MSA list server.
Any thoughts, suggestions, or recommendations for mail browsers better able to do the job?
More likely than not, the soap you use is the culprit. I find pure glycerine soap not drying nor chapping, even if used to excess. (In US, check your pharmacy for Nutrogena - UNSCENTED, or the same thing under the pharmacy name but at 1/2 price.)
Most "hand lotions" contain perfume in alcohol base and do more harm than good. At lunch, end of work, and night time, I work-in a good quantity of "A and D Ointment" - the variety WITHOUT ZnO. (Its customary use is to relieve diaper rash, and it can be found in baby-needs.)
By next morning, fingers are fine.
valdemar-at-fast.net No stock in either product.
---------- } From: James Martin {James.S.Martin-at-williams.edu} } To: microscopy-at-sparc5.microscopy.com } Subject: hand care for microscopists } Date: Thursday, October 30, 1997 8:40 PM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Here's a new thread. Hand care for microscopists. } } I spend most of each day preparing and analyzing samples using light } microscopy and FT-IR microscopy. Fingerprints are more than a nuisance, } so its frequent trips to the sink for a little soap and water. In the } winter I find my fingertips become dry and cracked. Lotion is out during } work hours. Cotton gloves shed linters. Finger cots provide some relief. } } Anybody have other suggestions? } } James "Fingers" Martin :) } Williamstown Art Conservation Center }
Cindy Bennett wrote: =================================================== I've got a question about plasma "etching" or "ashing" procedure to remove outer organic surfaces of polymer samples.
Another lab who did this for us in the past had a procedure where they did the etching in two steps: 1. oxygen plasma 2. argon plasma. (The other lab won't talk to us anymore, but that's another story...)
I can't figure out what what the argon plasma is supposed to do. The function of the oxygen plasma is clear: to oxidize the carbon and hydrogen, producing volatile substances. A mixed oxygen/argon plasma might make sense too, to control the ablation rate for instance. But the reason for doing pure oxygen and then pure argon eludes me.
Am I being dense? Can anybody give me a tip on this? (I'm a chemist, for goodness sake, but I still can't figure it out!) ================================================== Argon is used, in general, when one would want to remove a metal oxide and maybe some metals, cases where the aggressiveness of CF4 or other reactive fluorine based bases is not required. We are talking now about real "etching" and not just "plasma cleaning". I have heard of "formulas" where people have diluted oxygen with argon in order to moderate the etching of an organic surface. But I am unaware of any published literature that describes any benefit of first etching with pure oxygen and then etching with pure argon. Indeed, if the objective was to etch a polymer surface to reveal structure and morphology, to then expose it to argon would, at least to me, seem to be counter productive. The only exception I can recall was in the study of aluminum lithographic printing plates, the polymer emulsion layer was first etched off with oxygen and then the oxide layer was etched somewhat with argon to better "open" up the pore structure. The aluminum oxide was being "etched" slightly with the Ar.
I always hate to second guess another laboratory without hearing their rationale for doing something in some particular way. Here is the USA, an independent laboratory, especially one operating under ISO Guide 25 guidelines, and offering such a service would be expected to answer such a question in their report to the client. Since ISO Guide 25 had its origins in your corner of the world, I am surprised that laboratories there are not operating to the same standard?
Disclaimer: SPI manufactures a plasma etcher and would have an interest in seeing more people using this technique in microscopy. More information can be found on our website below.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
The stereo microscope is using only one of the optical paths to form the image on the film plane. Thus you will notice some side-to-side movement of the image as you focus the microscope. If your depth of field is limited and the subject is flat on the microscope stage (or base plate), only the center of the image will be in focus, the left and right edges being above or below the plane of focus. } -----------------------------------------------------------------------. } } Hi, there: } } I need some help on taking picture using a Zeiss STEMI SR. Some how the } image in the camera is not focused uniformly. The image (and therefor the } picture) is best focused at the central spot, and becomes blur at the } edges, most noticeably on one side. When I raise or lower the scope (or } stage), the image kind of move laterally, not up and down. I could not } figure out why, except suspecting the alignment of the scope may be off. } Any suggestions and comments are welcome. } } Jim
Russell E. Cook Scientific Associate Electron Microscopy Center for Materials Research Argonne National Laboratory Building 212 9700 South Cass Avenue Argonne, IL 60439 (630)252-7194 FAX: (630)252-4798
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
RE} EM: plasma etching procedure 10/31/97
I only reason I can think of for a final pure Ar plasma is to purge the chamber and pumping system of all pure oxygen prior to venting. I'm not aware of anyone else who does this, but it might be a safety concern for somebody.
********************************************************** Jake Schaper Product Analysis Lab Motorola, Inc. 1300 N. Alma School Rd. Chandler, Arizona 85224 Mail Drop CH240 Phone 602-814-4756 **********************************************************
--------------------------------------
Hello wise ones,
I've got a question about plasma "etching" or "ashing" procedure to remove outer organic surfaces of polymer samples.
Another lab who did this for us in the past had a procedure where they did the etching in two steps: 1. oxygen plasma 2. argon plasma. (The other lab won't talk to us anymore, but that's another story...)
I can't figure out what what the argon plasma is supposed to do. The function of the oxygen plasma is clear: to oxidize the carbon and hydrogen, producing volatile substances. A mixed oxygen/argon plasma might make sense too, to control the ablation rate for instance. But the reason for doing pure oxygen and then pure argon eludes me.
Am I being dense? Can anybody give me a tip on this? (I'm a chemist, for goodness sake, but I still can't figure it out!)
Message-Id: {3.0.2.16.19971031143030.425fbb24-at-inka.mssm.edu} X-Sender: massimo-at-inka.mssm.edu X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.2 (16)
At 01:20 AM 11/1/97 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I use Eudora as my email system, so I don't know if the following will work with Internet Mail & News.
In Eudora you can set filters which can look at any part of the message (header, sender, body, etc.) and take one of several actions (e.g. send to a specified mailbox). I have set up a mailbox "microscopy" and a filter which looks in the body of each incoming message for the prologue which is added by the Microscopy list server, in particular for the words "Microscopy ListServer". If these words are found, the message is automatically sent from the "In" mailbox to the "microscopy" mailbox. Your system should have an equivalent feature. If not, I suggest you take a look at Eudora (I have no financial interest in the company, I just like the product).
Sincerely,
Massimo Sassaroli _____________________________________________________ Massimo Sassaroli, D.Sc. Department of Physiology and Biophysics Box 1218 Mount Sinai School of Medicine One Gustave L. Levy Place New York, NY 10029-6574
A rapidly growin OEM of surface inspection equipment specializing in the development of state of the art micropscopy techniques is in need of a goal orientated leader to direct all production development and ensure that goals & objectives of plans are met within prescribed time frames and funding parameters. Requirements: BS & 10 years of related management experience. Diverse technical background, emphasis in optics and confocal microscopy. Experience: managing mutidiciplinary projects, intellectual property, team environment. Send resume to Kovex Corporation, 3711 Lexington Avenue N., Shoreview, MN. 55126 FAX: (612) 486-9785 or e-mail: kovex-at-spacesmar.net
} I am thinking of getting an IR viewing system for our SEM chamber. Any } advice or true to life adventures you can share?
We have a GW Chamberscope that uses infrared illumination attached to our Hitachi 2460N variable pressure SEM. Excellent system, extremely useful, no problems. I don't know how we got along without one. One suggestion: if you can, adjust the point of view to permit you to nearly look directly down on the sample (as is done with the Topcon) to better correlate to the SEM view.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
I have a question about the importance of cover slip thickness. Namely, how important is it to use a cover slip thickness for which the objective is designed? For example, if I'm using a 40X, NA 1.3 oil immersion objective which has the number 0.17 on it (corrected for a 170 micron cover slip thickness), what would be the effect on image quality if I used a cover slip with, say, a 300 micron thickness?
Also, what is the definition of "working distance?" I understand it to be the distance between the top of the cover slip and the lens of the objective, but I want confirmation of this definition.
Thank you very much,
Brian Haab U.C. Berkeley bhaab-at-zinc.cchem.berkeley.edu
Dan Kremser dkremser-at-levee.wustl.edu =============================================================
JOINT FALL MEETING
MISSOURI-ILLINOIS-KANSAS MICROBEAM ANALYSIS SOCIETY & CENTRAL STATES MICROSCOPY SOCIETY
Washington University Conference Center, St. Louis, Missouri
November 14, 1997
PROGRAM:
Chair: Lou Ross University of Missouri-Columbia
8:30-9:00 Registration, Vendor Display Setup, and Welcome Coffee.
9:00-9:10 Welcome
9:10-9:30 Rebecca Morlando 3M Company, Columbia, Missouri. "SEM to Confocal for Measuring Vias in Flexible Circuits"
9:30-10:00 Jingyue (Jimmy) Liu Analytical Sciences Center, Monsanto Company,St. Louis, MO "Resolution and Surface Sensitivity in Low Voltage SEM"
10:00-10:20 Dan Kremser Department of Earth and Planetary Sciences, Washington University, St. Louis, Missouri. "Coupling Laser Ablation with a Sector Field ICP-MS: Calibration Enhancement through the use of Smithsonian Microprobe Standards"
10:20-10:40 Morning Break---Coffee We thank Ed Immer/Electron Microscopy Sciences for support for our refreshments this morning.
Chair: Don Parker Briem Engineering, St. Louis, Missouri
10:40-11:40 Greg Meeker, US Geological Survey-Denver MAS tour speaker who will present the Chuck Fiori Memorial Technology Presentation.
"Standards for X-ray Microanalysis: How We Get Them, How We Use Them & Why We Will Always Need Them"
11:50-1:00 LUNCH See menu and details below on this page.
Chair: Mike Veith Department of Biology, Washington University
1:00-1:20 William Randle Missouri State Highway Patrol, Jefferson City,MO. "A Microchemical Test for Alkylamines of Water Gel Explosives"
1:20-1:40 L. Shannon Holliday Renal Division, Washington University Medical School, St.Louis, Missouri. "Use of Light, Electron and Confocal Microscopy in the Study of Osteoclastic Bone Resorption"
1:40-2:00 Gayleen Cochran Plant Biology Department, Southern IllinoisUniversity-Carbondale. "Spermatogenesis in Equisetum; a Correlated TEM and Fluorescence Microscope Study"
2:00-2:20 Angel R. Maden Plant Biology Department, Southern IllinoisUniversity-Carbondale. "Comparative Ultrastructure of Lycopidiaceae Spermatozoids"
2:20-2:40 Afternoon Break---Refreshments
2:40-3:00 Cheryl A. Nickerson Department of Biology, WashingtonUniversity, St. Louis, Missouri. "Histological Effect of Salmonella Infection on Murine Tissue"
3:00-3:20 Robert Mark Friedman Analytical Sciences Center, Monsanto Company, St. Louis, Missouri. "Imaging and Analysis with Time-of-Flight Mass Spectrometry and Related Techniques" *
3:30- Business Meetings
*
LUNCH DETAILS
Lunch will be a Deli Buffet consisting of Shaved Ham, Turkey, Roast Beef, American and Swiss Cheeses, various sandwich toppings, assorted Breads, Fruit Salad and Pasta Salad. Drinks and Desserts are included. The cost is $5.00 per person. Local eateries are located a short walking distance from the Conference Center for those who would rather dine offsite.
We need confirmation of those who plan to eat the Deli Buffet, By 10:00 AM, Tuesday November 11. Please contact Dan Kremser by phone (314-935-5605) or email (dkremser-at-levee.wustl.edu). Please confirm even if you have done so on the earlier mailing request. -- *************************** Lou Ann Miller Microscopic Imaging Lab College of Vet. Medicine University of Illinois 2001 S Lincoln Ave Urbana,Illinois 61801 217-244-1566 lamiller-at-ux1.cso.uiuc.edu
Microscopy Home Page: http://www.cvm.uiuc.edu/MicImagLab/MicImagLab.html
Central States Microscopy Society http://www.cvm.uiuc.edu/HomePages/LouAnnMiller/CSMS/csms.html
Personal Home Page: http://www.cvm.uiuc.edu/HomePages/LouAnnMiller/LAM.html
Another excellent product for hand repair is Extra Emollient Night Cream by Mary Kay. A jar costs $11 and lasts for a very long time. A physician friend of mine uses it and it has healed horrible skin cracking caused by latex allergy. Mary Kay consultants can be found either on the Web at www.marykay.com or in the white pages of the phone book.
} -----Original Message----- } From: Sara Miller [SMTP:saram-at-acpub.duke.edu] } Sent: Friday, October 31, 1997 11:22 AM } To: James Martin } Cc: microscopy-at-Sparc5.Microscopy.Com } Subject: Re: hand care for microscopists } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } ---------------------------------------------------------------------- } -. } } We have ethe same problem since we handle clinical samples and are } constantly washing with antiseptic soap. } } Sleep in gloves with lots of Vaseline on your hands. In the day time, } } chapstick applied just to the edges of the cracks (not all over your } hands) can help if cracks are not on the very tips. At least you can } apply it at lunch, etc. You can glue cracks together with superglue } (a } hint from my surgeon friend). Eucerine is a good hand cream } recommended } by my pharmacist; you might also try bag balm (for cows utters, } available } at farm supply stores) for times when you aren't at work. } } Sara Miller } } On Thu, 30 Oct 1997, James Martin wrote: } } } Date: Thu, 30 Oct 1997 20:40:20 -0500 (EST) } } From: James Martin {James.S.Martin-at-williams.edu} } } To: microscopy-at-sparc5.microscopy.com } } Subject: hand care for microscopists } } } } } ---------------------------------------------------------------------- } -- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } } ---------------------------------------------------------------------- } -. } } } } } } Here's a new thread. Hand care for microscopists. } } } } I spend most of each day preparing and analyzing samples using light } } microscopy and FT-IR microscopy. Fingerprints are more than a } nuisance, } } so its frequent trips to the sink for a little soap and water. In } the } } winter I find my fingertips become dry and cracked. Lotion is out } during } } work hours. Cotton gloves shed linters. Finger cots provide some } relief. } } } } Anybody have other suggestions? } } } } James "Fingers" Martin :) } } Williamstown Art Conservation Center } } } } Sara E. Miller, Ph. D. } P. O. Box 3020 } Duke University Medical Center } Durham, NC 27710 } Ph: 919 684-3452 } FAX: 919 684-8735
Vlademar et al: I have written two rules which cover 90% of all MSA mail. Rule 1) When Event is "new Item" type is "mail" contents are "TO: "microscopy" Move to Folder MSA. Rule 2) same as above except contents are "CC: "microscopy" " Move to Folder MSA. I hope this helps. bob m
On Fri, 31 Oct 1997, David S. Wexler, Ph.D. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hi, } } I have a question about the importance of cover slip thickness. Namely, } how important is it to use a cover slip thickness for which the } objective } is designed? For example, if I'm using a 40X, NA 1.3 oil immersion } objective which has the number 0.17 on it (corrected for a 170 micron } cover slip thickness), what would be the effect on image quality if I } used a cover slip with, say, a 300 micron thickness?
It is important. You are using a precision optical instrument and the cover slip is an indespensible part of the optical correction. Since you are using an oil immersion objective at 40 X (with oil I hope) it appears you want as much defined and clear detail as posible. Using any lense above about 40 X you can see the difference between #1 cover slips (0.13 to 0.17mm thick), #1 1/2 coverslips (0.16 to 0.19mm thick), and #2 cover slips (0.17 to 0.25mm thick). Now having said that, there is an assumption implicit in that. That is that the sample adhears directely to the underside of the coverslip. So don't "flood" with mounting medium.
As an asside, I know one microscopist who measures his coverslips with a micrometer and only uses the ones which are actually 0.17mm.
} Also, what is the definition of "working distance?" I understand it to } be the distance between the top of the cover slip and the lens of the } objective, but I want confirmation of this definition.
You are basically correct.... to be a "sticler" it is the nearest part of the lense, not any optical center of it.
} } Thank you very much, } } Brian Haab } U.C. Berkeley } bhaab-at-zinc.cchem.berkeley.edu }
Good luck,
Shalom from Jerusalem, Azriel
******************************************************** Azriel Gorski, Head azrielg-at-cc.huji.ac.il Optical Microscopy Laboratory Division of Identification and Forensic Science Israel National Police Jerusalem ISRAEL
Dear James, Most microscopists have white, nylon gloves used for handling high vacuum parts, which also cannot tolerate fingerprints. They are comfortable and breathe and do not shed much lint. Try an EM catalogue. You wrote: } } } Here's a new thread. Hand care for microscopists. } } I spend most of each day preparing and analyzing samples using light } microscopy and FT-IR microscopy. Fingerprints are more than a nuisance, } so its frequent trips to the sink for a little soap and water. In the } winter I find my fingertips become dry and cracked. Lotion is out during } work hours. Cotton gloves shed linters. Finger cots provide some relief. } } Anybody have other suggestions? } } James "Fingers" Martin :) } Williamstown Art Conservation Center } Regards, Mary
With all due respect to the members of this Listserver, I do feel compelled to provide additional insight into plasma processing and its related effects on TEM specimens. This response is largely due to Dr. Nestor Zaluzec's recent posting.
Both previous and recent experimentation clearly indicates effective cleaning times for TEM specimens of as little as 15 seconds. This factor is highly dependent on the type of plasma system used, since all plasma types are not equivalent. For those of you that are interested, I would suggest that you reference a book written by Michael A. Lieberman and Allan J. Lichtenberg entitled "Principles of Plasma Discharges and Materials Processing". It provides a great deal of insight into the various types of plasma and their applications.
Regarding the gas type and concentration, we have conducted quantitative measurements of contamination rates using Parallel Electron Energy Loss Spectrometry which indicates equivalent performance of a one minute processing time using a 25% oxygen 75% argon mixture, to a 5 minute cleaning with argon followed by a 5 minute cleaning with 100% oxygen.
When one considers input power into the plasma, whether it is DC or high frequency (HF), other factors such as chamber size and configuration, gas type, plasma processing chamber pressure, electrode style and placement, and Faraday shielding also provide significant contributions to the plasma physics (ion energy, plasma potential, plasma density, electron temperature, and floating potential).
In an non-equilibrium, inductively coupled plasma, increasing the high frequency input power actually decreases the ion energy, which is the most critical parameter for optimal plasma processing. When the proper execution of plasma formation occurs, ion energies of {20 eV are achieved which results in negligible specimen heating and no sputtering effects either from or onto the specimen. On the reverse side, both sputtering and oxygen ion implantation are very real possibilities when the ion energies become excessive.
When considering the safety issues associated with pure oxygen, let us not forget the utilization of pressure regulators certified for oxygen usage, and the avoidance of oils in the pressurized portion of the system.
To conclude, I would be happy to discuss both any data that we have obtained to date, and anyone's issues related to the plasma processing of EM specimens. At this juncture, I do believe that it would be most appropriate to conduct these activities off the Listserver by contacting me directly as follows.
Thank you for your consideration.
Best regards,
Paul
Paul E. Fischione, President E.A. Fischione Instruments, Inc. 9003 Corporate Circle Export, PA 15632 Phone 412-325-5444 FAX 412-325-5443 paul.fischione-at-internetmci.com www.fischione.com
We have GW Electronics chamber cameras on Hitachi and Jeol SEM's. They work great. Just remember to turn the LED illumination off for doing EDX work, as they affect the EDX detector.
We seek information on atomic force microscopes for a future purchase.
Specifically information from:
(a) vendors: models available, capabilities and specs, housing requirements, pricing, references of users, etc.
(b) individual users: models to avoid, positive experiences with a particular model, maintenance, software available, etc.
I shall compile this information for others who may have similar interests. Thank you.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing 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 ####################################################################
Does anyone have suggestions concerning how to prepare a sample of paper laminated with Latex and Saran for examination by SEM/EDX, optical microscopy and micro FTIR. Ideally I would like to observe the sample in cross section with a surface similar in quality to those we prepare by metallographic techniques. Simply cutting by razor smears the surface. The sample was still ductile while submerged in LN2 so we could not obtain a fracture surface. We have had some success cutting with razor while under LN2 but there are still signs of smearing. We could microtome under LN2. Does anyone have other suggestions concerning sample preparation? Are metallographic techniques (embed, grind and polish) feasible? The embedding material must not affect the sample.
Thanks in advance,
Ed Kurz Institute of Materials Science, U-136 University of Connecticut 97 North Eagleville Road Storrs, CT 06269-3136 ekurz-at-mail.ims.uconn.edu (860) 486-4186 phone (860) 486-4745 fax
I think the positioning of the camera depends on the reason you have for using the camera. I have mine looking parallel to the axis of stage tilt, just below the bottom of the pole piece. I can see the gap between my sample and the pole piece as I raise and tilt the sample. I can also see the proximity of the various detectors to the sample. This saves on destroyed samples and detectors!
A young student working in my lab want to study some liposomes using negativ staining. So far with no luck. So if someone out there could help, we would be most grateful. Up til now she has used 2 %PTA pH 7.6 on formvar film coated with carbon. A suspension of liposomes is added on top of the grid, excess liquid removed followed by PTA. We have so far seen only the biggest liposomes, but it seems to be difficult to 'catch' the solicited solution of small particles.
Tips n' tricks would be greatly appreciated.
Best wishes Randi Olsen
Department of Electron Microscopy Faculty of Medicine University of Tromsoe, Norway
Dear all. We are in the lucky situation in my department to have 'spare' money this year, and want to replace our 'old' LKB Tissue Processor who 'died' earlier this fall. It seems like the new RMC or the Lynx machine are the two most current ones. It would be of great help if anyone would share their experiments with us. I'm especially interested in the reliability of the plastic tubes for the processors. We've had lots of problems with the LKB because of lack of accuracy of the plastic ware.
Thanks in advance.
Randi Olsen Department of Electron Microscopy Faculty of Medicine University of Tromso Norway
Hi Randy...we have one of the RMC processors which is about one generation old. Ours allows one to pre-cool (or warm) the solution tube in which your sample will go into next, as well as the current tube. The new machine only controls the temperature of the current tube (which has never made sense to me).
We like our machine and have had few problems. We also had one of the previous machines with the LKB nameplate (they were both made by the same company), and the tubes, with very thin walls, would bend and cause the specimen containing stacking rings to get stuck and left behind. This would raise havoc in the machine, with samples often getting mixed up. The RMC tubes are thicker, and we have not experienced a return of the problem. We are careful to put holes in the caps when the solution tube contains a highly evaporative solvent such as propylene oxide. We found that the vapor pressure would dislodge the caps and cause them to go askew. They would then not be lifted off correctly and would topple into the machine, invariably causing a jam.
Ed Kurz asked: ================================================ Does anyone have suggestions concerning how to prepare a sample of paper laminated with Latex and Saran for examination by SEM/EDX, optical microscopy and micro FTIR. Ideally I would like to observe the sample in cross section with a surface similar in quality to those we prepare by metallographic techniques. Simply cutting by razor smears the surface. The sample was still ductile while submerged in LN2 so we could not obtain a fracture surface. We have had some success cutting with razor while under LN2 but there are still signs of smearing. We could microtome under LN2. Does anyone have other suggestions concerning sample preparation? Are metallographic techniques (embed, grind and polish) feasible? The embedding material must not affect the sample. =================================================== We have been cutting these kinds of samples for some years and have come to the conclusion that there is only one "right" way! I am assuming you have a "paper" substrate, one impregnated with latex and the whole layer is laminated to a layer of "Saran". If the goal is to obtain the "perfect cross-section" then this can be done only via the methods of ultramicrotomy, using cryo techniques, and a diamond knife.
Our step by step procedure would be the following: 1] Sputter coat gold (or preferably Pt but gold is OK) on both sides, the paper side to keep the embedding resin from interacting and possibly dissolving some component in the paper substrate, or even swelling it and the Saran (PVDF) side to act as a superb "decorator" to show the Saran/embedding media interface.
2] We would recommend our own SPI-Pon(TM) 812 Epoxy Embedding Resin but a good number of the other readily available Epon (R) 812 "substitute" materials should work just as well, ones offered by other EM supply firms. Epon Araldite(R) and perhaps other resin systems will "work" too, but the "812" system seems to result in being able to obtain what you want in the shortest possible period of time in terms of learning curve development.
3] Because paper has inorganics in it (e.g. clays, etc.), you surely don't want to use a brand new life science knife to do the cutting. We would recommend saving money and purchasing a materials science diamond knife for this purpose. However, the availability of a beat up life science knife, in need of resharpening, might be quite satisfactory (the quality of the knife edge need not be to the standard as if you were to be looking at the sections, but if the knife is too far gone, you will pick up an intolerable number of defects on the final block you are going to examine.
4] In your case the sections would be thrown out. This brings tears to our eyes since it has been our experience, for these kinds of laminated samples, that the TEM results often times contain much more useful information than the SEM results! But the "faced-off-block" is now the ideal sample for insertion into the SEM (after some metallization or carbon coating).
5] When you look by SEM, you will not have any distortion or interaction of the resin with the sample itself. You might be disappointed in the contrast and to improve it, you might give it a 30 second oxygen plasma etching exposure, so that the gold lines stand up in "relief" better highlighting the location of your sample. If the latex is a polyBD type, then you can expose the sample to osmium tetroxide which could provide additional contrast to the sample, at least in terms of the penetration of the latex.
6] If you have either a "stone" or "fisheye" in either of the layers, if you have successfully cut through the center of the defect, then it will be ideally exposed for EDS, LM, or micro FT/IR analysis. Unlike the case if you were to look at the lateral surface, in this instance, the defect is not being covered up with anything.
This really is a straight forward procedure and has literally "worked" on numerous samples of this and similar types.
Disclaimer: SPI Supplies offers embedding resins, materials science diamond knives, and plasma etchers for doing this kind of work. So we would have a vested interest in the promotion of this kind of approach.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
In a message dated 97-11-03 20:57:11 EST, you write:
{ { Subj: TEM Liposomes. Neg.staining. From: Randio-at-fagmed.uit.no (Randi Olsen)
Dear fellow microscopists.
A young student working in my lab want to study some liposomes using negativ staining. So far with no luck. So if someone out there could help, we would be most grateful. Up til now she has used 2 %PTA pH 7.6 on formvar film coated with carbon. A suspension of liposomes is added on top of the grid, excess liquid removed followed by PTA. We have so far seen only the biggest liposomes, but it seems to be difficult to 'catch' the solicited solution of small particles.
Tips n' tricks would be greatly appreciated.
Best wishes Randi Olsen
Department of Electron Microscopy Faculty of Medicine University of Tromsoe, Norway
} }
I have a couple of suggestions:
1] Apply your suspension to the Formvar surface of the support film rather than the carbon surface. You may 'catch' more of the particles that way.
2] Treat your support films with POLY-L-LYSINE before applying the liposome suspension. To do this use a .01% aqueous solution of POLY-L-LYSINE.
Apply a drop to the carbon surface of the support film and after 5 mins remove excess liquid.
Allow another five minutes for the surface to dry thoroughly (can be speeded up by placing in an oven at 40 degrees centigrade for one minute).
Now apply your suspension and carry out the negative staining as before.
I am finding that the most stubborn material can be examined using this process.
Some time ago I asked for advice on how to tackle our problem with soft resin (see the following e-mail). I received numerous helpful hints and advice which I am very thankful for. One particular advice I was given (by Dr Bronwen Cribb, CMM, UQ) was to replace the alcohol in dehydration series with acetone. Apparently, a small amount of alcohol gets trapped in the block (why??) which causes some blocks to remain soft at the end. The first series of specimens we did with acetone did not show any problem. I thought it was worth sharing this with everybody.
Majid
+++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Dear All
We are doing TEM on cultured Koala lymphocytes. We are having an on going problem with Spurr's resin (we use medium Spurr's). In some cases resin doesn't get polymerised and stays soft even after 3-4 days in sixty degree oven. The interesting point is that it doesn't happen with every sample. Even in a series of different samples which are processed at the same time and embeded in the same batch of resin, some remain soft while the others are quite Ok.
We heard that some components of culture media may interfere with resin polymerisation so we have been careful to wash the cultured cells properly but it did not eliminate the problem. Any advice on how to tackle the problem is highly appreciated. I would also be thankful if you suggest a way to revive those samples which are embeded in soft resin.
Regards
M. Ghoddusi
Majid Ghoddusi Division of Veterinary Pathobiology The University of Queensalnd QLD 4072 Australia
azriel gorski wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } On Fri, 31 Oct 1997, David S. Wexler, Ph.D. wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } -----------------------------------------------------------------------. } } } } Hi, } } } } I have a question about the importance of cover slip thickness. Namely, } } how important is it to use a cover slip thickness for which the } } objective } } is designed? For example, if I'm using a 40X, NA 1.3 oil immersion } } objective which has the number 0.17 on it (corrected for a 170 micron } } cover slip thickness), what would be the effect on image quality if I } } used a cover slip with, say, a 300 micron thickness? } } It is important. You are using a precision optical instrument and } the cover slip is an indespensible part of the optical correction. Since } you are using an oil immersion objective at 40 X (with oil I hope) it } appears you want as much defined and clear detail as posible. Using any } lense above about 40 X you can see the difference between #1 cover slips } (0.13 to 0.17mm thick), #1 1/2 coverslips (0.16 to 0.19mm thick), and #2 } cover slips (0.17 to 0.25mm thick). Now having said that, there is an } assumption implicit in that. That is that the sample adhears directely to } the underside of the coverslip. So don't "flood" with mounting medium. } } As an asside, I know one microscopist who measures his coverslips with a } micrometer and only uses the ones which are actually 0.17mm. } } } Also, what is the definition of "working distance?" I understand it to } } be the distance between the top of the cover slip and the lens of the } } objective, but I want confirmation of this definition. } } You are basically correct.... to be a "sticler" it is the nearest part of } the lense, not any optical center of it. } } } } } Thank you very much, } } } } Brian Haab } } U.C. Berkeley } } bhaab-at-zinc.cchem.berkeley.edu } } } } Good luck, } } Shalom from Jerusalem, } Azriel } } ******************************************************** } Azriel Gorski, Head azrielg-at-cc.huji.ac.il } Optical Microscopy Laboratory } Division of Identification and Forensic Science } Israel National Police } Jerusalem } ISRAEL
OK, but wait a minute. Brian also said he is using oil immersion, as well he should be with the lens he described. Oil immersion is sometimes known as "homogeneous immersion" because the optical medium from (and including) the objective front element all the way through to (and including) are of homogeneous optical character. In other words, they are of the same refractive index. The immersion oil is of the same RI as the objective front element, the same as the cover slip, the same as the slide, the same as the condenser front element and (usually) about the same as the mounting medium. Thus, in the special case of oil immersion, it is generally held that the cover slip thickness is irrelevant within reasonable bounds. This is why very often on oil immersion objectives there is no indication of best cover slip thickness. Where cover slip thickness is important is with high numerical aperture _dry_ lenses. The cover slip is always an integral part of the optical system and, as such, its characteristics (including thickness) are incorporated in the calculations of the entire objective lens system. But with high NA lenses there is less tolerance for the aberrations introduced by improper cover slip thickness. Here is an excerpt from the Particle Atlas Electronic Edition (You will note a slight difference in opinion from mine of c.s. thickness with oil immersion):
Chapter 1. Optics and the Microscope Section D. Compound Microscope Subsection 1. Objectives Subsection g. Cover slip thickness
"An important consideration in using different objectives is cover slip thickness. Just how important is the cover slip in the formation of sharp microscopical images? Spinell and Loveland answer this question very completely in a paper entitled "Optics of the Object Space in Microscopy." For the average skilled microscopist, cover slip thickness should be close to the recommended thickness of 0.17 mm (continental and oriental microscopes) or 0.18 mm (U.S. and British microscopes). The allowable variation before detectable image deterioration occurs depends on the numerical aperture of the objective: it is +/- 8 micrometers for a 0.95 NA dry objective; +/-15 micrometers for 0.85 NA; +/-45 micrometers for 0.65; for objectives of NA less than or equal to 0.25 it makes little difference whether a cover slip is used or not. It is also best to use a cover slip of correct thickness for immersion objectives.
"Cover slips vary greatly in thickness and some manufacturers' cover slips are better than those of others. In one test reported by Loveland, a group of Corning No. 1-1/2 cover slips averaged 184 micrometers in thickness and about half of the slips were suitable for use with the 0.95 NA dry objective; nearly 95% were suitable with a 0.85 NA objective (assuming that the objectives are corrected for use with 0.18 mm cover slips). It is best to buy No. 1-1/2 cover slips and to micrometer them when doing critical work."
(The cited Spinel and Loveland paper will be found at J. Roy. Micros. Soc., 79:59-80, 1960.)
By the way, IMHO, Brian's question well illustrates why it is deplorable that in modern universities there is usually no place for a student to learn the fundamentals of light microscopy. Maybe now that you can actually spend $100K on a light microscope, and now that confocal and other "modern" techniques are being used in genetic and other intensely popular research areas, science may once again begin to take the light microscope seriously. Perish the thought, but could we actually see some thought being given to serious instruction on how to use the instrument? There's a radical notion!
(Sorry, its late at night and I'm tired and grumpy. :-p No fault of yours Brian. How could you know better when you've nowhere to turn for education on the subject.)
--
Stephen A. Shaffer MicroDataware sshaffer-at-microdataware.com (business) http://www.microdataware.com (temporarily out of service) steve_shaffer-at-compuserve.com (personal) http://ourworld.compuserve.com/homepages/steve_shaffer/
1. Procedure: I float my grids on top of a drop of solution on Parafilm in a wet chamber for 10-15 min., then pick the grid and blot out the excess solution with Whatman 1 filter. I transfer on the stain solution for 30 sec., blot and dry. In my hands this works better than the other way (solution on grid) which seems to favor uneven spread of the material. 2. Dilution of the liposome solution: find the right dilution which will allow sufficient dispersion of the material on the grid. With a diluted solution you may have to search a little more but there is a far better chance to see the full range of sizes present in your sample. This should prevent the big guys to mask the little ones. 3. Stains: I have used both UAc 2% / trehalose 1% in ddH2O or PTA 2% / trehalose 1% in ddH2O (according to J. Robin Harris) with some success.
I hope this helps. Best regards, Michel
**************************************************** Michel Deschuyteneer, Ph.D. deschuyt-at-sbbio.be Scientist Electron Microscopy Laboratory
SmithKline Beecham Biologicals Rue de l'Institut, 89 B1330 Rixensart, BELGIUM Tel: +32-2-656 9290 Fax: +32-2-656 8164 **************************************************** Standard disclaimer: the opinions expressed in this communication are my own and do not necessarily reflect those of SmithKline Beecham. ****************************************************
Dear all Just my view. I have been using Pegasus mail. The latest version is very versatile and free. Have two different options in the filtering which will look at closed and open folders, with option for including and excluding certain mail if certain key words appear in either the text, heading or subject field. I am a mere happy user. } } } } Has anyone figured out how to segregate from other mail the e-mail } } originating in this forum? } } } } I use Internet Mail & News (v4.70) bundled with Microsoft IE 3.02. The } } "Inbox Assistant" in "Mail" menu is meant to assist in automatic } } segregation to sub-folders, but I have been unable to make it work with } } stuff from the MSA list server. } } } } Any thoughts, suggestions, or recommendations for mail browsers better able } } to do the job? } }
} } } } I use Eudora as my email system, so I don't know if the following will work } with Internet Mail & News. } } In Eudora you can set filters which can look at any part of the message } (header, sender, body, etc.) and take one of several actions (e.g. send to } a specified mailbox). } I have set up a mailbox "microscopy" and a filter which looks in the body } of each incoming message for the prologue which is added by the Microscopy } list server, in particular for the words "Microscopy ListServer". If these } words are found, the message is automatically sent from the "In" mailbox to } the "microscopy" mailbox. } Your system should have an equivalent feature. If not, I suggest you take a } look at Eudora (I have no financial interest in the company, I just like } the product). }
John, the majority of the makers of AFM systems can be found on the Web and they provid a good background to the instruments they sell. Namely ...
Digital Instruments - www.di.com Topometrix - www.topometrix.com Park Scientific (now part of thermospectra) - www.park.com
These are the three main companies, through Burleigh now make a research grade AFM as well as low priced personal AFM and STMs. DME a danish company make the Rasterscope which is a resonably priced machine with mid range capabilities. Recently on the market are Molecular devices and Tools which are a Russian company - i haven't seen their machine in action - but a contact email address for the states is howard-at-ktecintl.com.
As for which machine is the best - I would say that would depend to a degree on you application and the amount of money you wish to spend. Some of the companies' research instruments are for example better for working in liquid than others, others allow for larger samples. I would recomend you get a demonstatration by as many of the manufacturers as you can before making any decision.
Dr. Giles Sanders Laboratory of Biophysics and Surface Analysis School of Pharmacy Nottingham University England
On Mon, 3 Nov 1997 13:37:39 -0500 Ed Kurz {ekurz-at-mail.ims.uconn.edu} wrote:
} Does anyone have suggestions concerning how to prepare a sample of paper } laminated with Latex and Saran for examination by SEM/EDX, optical } microscopy and micro FTIR. Ideally I would like to observe the sample in } cross section with a surface similar in quality to those we prepare by } metallographic techniques. Simply cutting by razor smears the surface. } The sample was still ductile while submerged in LN2 so we could not obtain } a fracture surface. We have had some success cutting with razor while } under LN2 but there are still signs of smearing. We could microtome under } LN2. Does anyone have other suggestions concerning sample preparation? Are } metallographic techniques (embed, grind and polish) feasible? The embedding } material must not affect the sample. } } Thanks in advance, } I've had good results from embedding paper in epoxy resin and polishing, finishing with quarter micron alumina. I also tried LR White, which did not make such good blocks. It doesn't seem to make much difference whether or not you vacuum impregnate.
Regards, Eric ---------------------- Dr Eric E. Lachowski University of Aberdeen Department of Chemistry Meston Walk Old Aberdeen AB24 3UE +44 1224 272934 e.lachowski-at-abdn.ac.uk
We have a problem with our Zeiss DSM 962 which our engineers are, as yet, unable to solve. The turbo pump switches off and an error message indicates that there is "...no 220v supply or the supply line is interrupted". This supply has been checked and found to be normal.
The problem was initially intermittent, occurring immediately or several minutes after the instrument was switched on from cold. It also occurred occasionally after chamber evacuation. However, recently the problem has become so bad that the error message does not respond to the reset button and will only clear when the microscope is switched off at the rear.
The engineers at LEO suspect a fault on the circuit board relating to the vacuum system. I would be interested to hear directly from anyone who has had a similar experience or who might offer advice.
At 07:56 4/11/97 GMT0BST, Giles Sanders wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Also JEOL has a long experience with AFM and STM. Please contact your local sales office.
You can find us on our webside : http://www.jeol.co.jp or http://www.jeol.com
You might try floating the grid on a drop of the suspension on a piece of Parafilm or other hydrophobic surface. You can also try adding a wetting agent like Bacitracin. Or you could charge the surface of the grid with polylyine if the liposomes are negatively charged. I have always used 1-2% uranyl acetate for neg. staining of liposome. Maybe the suspension needs to be diluted as well.
It is possible that you only have large ones in your suspension despite assurances from whoever made them that they should be small. Have they been sized by light scattering or some other method?
Greg Erdos
At 11:57 PM 11/3/97 +0100, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America Scientific Director, ICBR Electron Microscopy Core Lab PO Box 118525 Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ Home of the #1 Gators ***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
For current users of automated tissue processors: I would be very interested to know processing protocols and any special resin formulation that is particularly suited for the processor. TIA
Walt Bobrowski Parke-Davis Research bobroww-at-aa.wl.com
} } Has anyone figured out how to segregate from other mail the e-mail } } originating in this forum? } } } } I use Internet Mail & News (v4.70) bundled with Microsoft IE 3.02. The } } "Inbox Assistant" in "Mail" menu is meant to assist in automatic } } segregation to sub-folders, but I have been unable to make it work with } } stuff from the MSA list server. } } } } Any thoughts, suggestions, or recommendations for mail browsers better able } } to do the job? } }
Sorry to post this to the list. But I did not read the original e-mail and the subsequent reply doesn't include the information to allow me to reply to the originator personally.
I use Microsoft IE 3.02 and the Internet Mail and News(v4.70) too. I have been able to segregate most of the MSA mailings using the Inbox assistant. You do need to use more than one instruction to take care of most of the possiblities. I use the following, which seems to filter over 95% of MSA messages into my "MICROSCOPY" box
Move to "MICROSCOPY" if "TO" contains Microscopy-at-Sparc5.microscopy.com Move to "MICROSCOPY" if CC contains Microscopy-at-Sparc5.microscopy.com Move to "MICROSCOPY" if "TO" contains Microscopy and Listserver
I get an occasional rogue MSA message that does not conform to the above but they are rare. It might be worth a try before going to the trouble of changing over your e-mail browser.
The Oklahoma Microscopy Society (OMS) will be holding its fall technical meeting in conjunction with the Oklahoma Academy of Science (OAS) at the University of Arts and Sciences of Oklahoma in Chickasaw, Oklahoma on Friday, November 7, 1997. Any and all are welcome to attend.
Key note speakers for this meeting include: (1) Dr. Biao Ding (Department of Botany, Oklahoma State University, Stillwater, OK) speaking on "Elucidating Intercellular Communications in Plants: Role of Microscopy" and (2) Dr. Robert Nordquist (Department of Ophthalmology, University of Oklahoma Health Sciences Center, Oklahoma City, OK) speaking on "The Ultrastructure of Laser/Tissue Interactions and Clinical Applications".
Other points of business:
-OMS will be celebrating its 20th Anniversary at the meeting.
-Students will be competing for the annual Timpano Award which is an all expense paid trip to the national MSA meeting the following year.
-Items regarding the upcoming joint spring meeting with the Texas Society will be discussed.
Any questions, please contact Ginger Baker (Secretary/Treasurer) at (918) 561-8232 or Phoebe Doss (President) at (405) 744-6765.
The Oklahoma Microscopy Society (OMS) will be holding its fall technical meeting in conjunction with the Oklahoma Academy of Science (OAS) at the University of Arts and Sciences of Oklahoma in Chickasaw, Oklahoma on Friday, November 7, 1997. Any and all are welcome to attend.
Key note speakers for this meeting include: (1) Dr. Biao Ding (Department of Botany, Oklahoma State University, Stillwater, OK) speaking on "Elucidating Intercellular Communications in Plants: Role of Microscopy" and (2) Dr. Robert Nordquist (Department of Ophthalmology, University of Oklahoma Health Sciences Center, Oklahoma City, OK) speaking on "The Ultrastructure of Laser/Tissue Interactions and Clinical Applications".
Other points of business:
-OMS will be celebrating its 20th Anniversary at the meeting.
-Students will be competing for the annual Timpano Award which is an all expense paid trip to the national MSA meeting the following year.
-Items regarding the upcoming joint spring meeting with the Texas Society will be discussed.
Any questions, please contact Ginger Baker (Secretary/Treasurer) at (918) 561-8232 or Phoebe Doss (President) at (405) 744-6765.
I need to prepare TEM specimens of Inconel 718 superalloy . I was going to start with perchloric and ethanol at about - 35 C. I would however appreciate getting "recipes" from people with more experience on this matter.
------------------------------------------------------------------------ The following material may be of interest to you. If it isn't, we apologize for any inconvenience. ------------------------------------------------------------------------
http://www.infowatch.net InfoWatch would like to introduce our complete Hosting and Promotional service.
FOR ONLY $29.95 PER MONTH !
* 30 MB (megabytes) of disk space * 1000 MB = 1 GB (gigabyte) of data transfer per month * 8 E-mail accounts * Access to your account anytime via the Web Control Panel * MS Front Page * Unlimited FTP updates * Personal CGI directory for your own scripts (or use ours) * Advanced web statistics analyzer program * E-mail forwarding, auto responders, and vacation reply * Domain name registration (www.yourname.com) * Register your domain with over 250 search engines - FREE! * Friendly customer support
* Inquire about our $24.95/month plan
If for any reason you are not satisfied with InfoWatch's service after 30 days, we will transfer you back to your original host and pay any transfer fees.
Marti, Jordi wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } After polishing my sample I immersed it in acetone to dissolve the cement } and after it had separated from the glass support I examined it by OM . } At that point I noticed that a thin solid film of the colloidal, } non-crystallizing silica, had formed on the surface of the sample. Is there } a safe way of removing this film ?? I've tried soaking the sample in the } colloidal suspension and in water but it did no seem to help. } } I suspect the film formed because I failed to wash the sample properly } before putting it in the acetone but I hate the idea of starting all over } again. } } I would appreciate any suggestions. } } Jordi Marti Dear Jordi,
Our Micro Organic Soap is used to remove our Non-Crystallizing Colloidal Silica Suspension in both 0.05 and 0.02 micron sizes. Either dilute it in water or use it full strength. The part number is 148-10000 and sells for 10 dollars a quart.
I have also heard Methanol works sometimes.
Good Luck,
Gary Liechty Product Application Specialist
Allied High Tech Products, Inc. 2376 E. Pacifica Pl. Rancho Dominguez, Ca. 90220 800-675-1118 310-635-2466 310-762-6808 Fax www.AlliedHighTech.com
Products for Materiallographic, SEM and TEM Sample Preparation
So how do you tell if you have spherical aberration due to cover glass thickness? You look at a tiny particle as you go in and out of focus. Spherical aberration will give you a small white spot in the center when you are out of focus one way, and not the other. Also the diffraction pattern around the particle will be different on one side of focus than the other--it is not symmetric in out-of-focus direction.
best regards mark
Mark W. Lund, PhD Director } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"The state is good at simple tasks, like killing people and seizing their wealth. It has far more trouble reaching inside individuals and making them good." Doug Bandow
A user in our lab wishes to image radiolarians (nearly pure SiO2) and separate them from the matrix (smectite to illite clay) in order to do an analysis of area/volume ratios.
In fractured samples we can see the radiolarians easily in the SEM, but he wants to be able to do image analysis on a picture and pick out the rads by doing some thresholding. In our pictures, the rads and the matrix are all similar in gray value.
Whole rads are 60 - 80 um in diameter, but they are also found as 'chunks' as small as 3 um. The clay particles of the matrix are from 1 - 5 um in size. He wants to separate even small particles of rads to get a measure of displacement and volume changes. I suggested that he may have to prepare a polished surface to examine, but he would like to avoid doing that since the samples are somewhat friable and he is not sure how to proceed.
We tried SEI and BSE imaging in the SEM, but the gray values are too close to do much thresholding. We tried doing some x-ray mapping, but Si is abundant in both the rads and the matrix so not much to go on there. Any elements unique to either the rads or the matrix seem to be present in concentrations too low to make a good separation using x-ray maps.
I have told him that I think this is a hard problem, but that I would send a message to the list to see if anyone had any ideas or leads on techniques. So, what do you think?
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
I would like to hear from anyone who has experience in the staining of poly(4-vinyl pyridine). The material in question is a copolymer diblock of poly(styrene-b-4-VP). If anyone out there can offer assistance I'd appreciate hearing from you. Thanks.
Paul Gerroir Xerox Research Centre of Canada Subscibe Microscopy Paul_Gerroir-at-xn.xerox,com
Good question: Not readily, I'm afraid. It is actually difficult to assess whether liposomes are uni- or multi-lamellar with neg staining. Actually, the best method to answer this is cryo-EM of a frozen hydrated suspension of the liposomes.
Regards, Michel
At 18:21 4/11/97 +0100, you wrote: } } Michel, } } I wonder how readily do you get to see if your liposomes are } multilamellar or not? } } Manoj } } Dr. Manoj MISRA, } Unilever Research } 45 River Road } Edgewater, NJ 07020 } (201)840-2702 (voice) } (201)840-8299 (fax) } Manoj.Misra-at-unilever.com } **************************************************** Michel Deschuyteneer, Ph.D. deschuyt-at-sbbio.be Scientist Electron Microscopy Laboratory
SmithKline Beecham Biologicals Rue de l'Institut, 89 B1330 Rixensart, BELGIUM Tel: +32-2-656 9290 Fax: +32-2-656 8164 **************************************************** Standard disclaimer: the opinions expressed in this communication are my own and do not necessarily reflect those of SmithKline Beecham. ****************************************************
I was asked recently if it is possible to get a final image on a Zeiss 902 that shows signs of astigmatism just in one half of the negative, the other half appears to be OK. Could this situation occur in a microscope that is OK due to misalignment of the energy filtering system by the user, or is this definitely a hardware problem?
Thank you for your help, Stefan
Dr. Stefan Hillmer Albrecht-von-Haller Institut fuer Pflanzenwissenschaften Universitaet Goettingen Untere Karspuele 2 37073 Goettingen Deutschland
Tel (+49) 551-392013 Fax (+49) 551-397833 e-mail shillme-at-gwdg.de
At 06:25 PM 11/4/97 -0600, Bob Wise wrote: } I'm considering buying some rebuilt SEM filaments (W). I am seeking input } from users as to how they compare to new (manufacturers or second source).
This issue comes up from time to time. Energy Beam Sciences has been manufacturing new and rebuilt tungsten filaments for EMs for more than 25 years, and I am regularly asked this question. I can't speak for other manufacturers, but I can state categorically that *our* rebuilt filaments are functionally identical to *our* new filaments. The bases are cleaned, identical filament loops are welded to the posts, and the finished filaments are aligned, vacuum annealed, then checked again for alignment. In blind tests, there was no variation at all between the performance of new and rebuilt filaments in the same microscope.
That being said, there may well be differences in *design* between filaments sold by the column manufacturer and rebuilt filaments manufactured by a third party. These differences can very well influence filament performance, both positively and negatively. If I were considering the use of rebuilt filaments, I would ask the manufacturer if the loop configuration differs from the filaments sold by the EM manufacturer and, if so, why.
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
Further to the "Dry hands" thread I've seen lately, I think there's a general humidity problem in a lot of labs, at least in colder climates where the central heating is on all winter. In my own lab, in a large building built in the late '70s, the humidity can get down to 15 - 20% by February or early March. This is probably a good environment for, say, preserving Lenin's corpse, but is not generally appropriate for living non-communist microscopists. I don't work with a lot of chemicals or irritants of any kind (except the Admin people), so when my hands get dry and chapped, I think it's just because of the dessicating environment. I'm thinking of installing a humidifier in the lab here, at least for winter use. Has anyone out there been able to successfully regulate a comfortable humidity level in a cool-climate lab? I'm going to have to be careful about approaching management to pay for a humidifier, though, because our building is just now going through some major renovation to take care of a minor outbreak of Stachybotras (a damp-loving mold that can be quite toxic to some people) that occurred in another, much moister part of the complex. So when they hear anything about me wanting to increase a humidity level somewhere, they may get a little squirrely on me.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Manoj,
It is well documented in literature that negative staining deteriorates the original structure of liposomes. The technique should not be used for their observation!!!!!!.
I always perform the LT observation in TEM of vitrified thin films in parallel with freeze fracture. Discrimination between uni- and multi-lamellar in thin films is straight forward. The freeze fracture in parallel is to have a confirmation of your findings in thin film. The thin film preparation can introduce various artefacts, e.g.: 1 selection of diameter size ( Determined by the film thickness the liposomes are ordered in their diameter. Diameters larger than the thickness of the film are excluded from the thin film and will not be visible in the TEM. 2.in case of viscous liquids, mechanical stresses are applied during thin film preparation, which can cause phase inversion. 3.Diameter sizes larger than the film thickness present in the film can be deformed and have lost their spherical shape (have become flattened between both air/water interfaces of the film. Tilting of the grid in the beam is needed to check on the shape of the vesicles. In the freeze fracture replica all sizes are present in their actual shape and distribution in the dispersion. Replication is limited in discrimination between uni- and multi-lamellar vesicles. Insight in this aspect is only obtained in case of cross-fractures through vesicles.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi Manoj.
Good question: Not readily, I'm afraid. It is actually difficult to assess whether liposomes are uni- or multi-lamellar with neg staining. Actually, the best method to answer this is cryo-EM of a frozen hydrated suspension of the liposomes.
Regards, Michel
At 18:21 4/11/97 +0100, you wrote: } } Michel, } } I wonder how readily do you get to see if your liposomes are } multilamellar or not? } } Manoj } } Dr. Manoj MISRA, } Unilever Research } 45 River Road } Edgewater, NJ 07020 } (201)840-2702 (voice) } (201)840-8299 (fax) } Manoj.Misra-at-unilever.com } **************************************************** Michel Deschuyteneer, Ph.D. deschuyt-at-sbbio.be Scientist Electron Microscopy Laboratory
SmithKline Beecham Biologicals Rue de l'Institut, 89 B1330 Rixensart, BELGIUM Tel: +32-2-656 9290 Fax: +32-2-656 8164 **************************************************** Standard disclaimer: the opinions expressed in this communication are my own and do not necessarily reflect those of SmithKline Beecham. ****************************************************
PHILADELPHIA SOCIETY FOR MICROSCOPY MEETING NOTICE: Wednesday Night, NOVEMBER 12, 1997
RESOLVING CHEMISTRY WITH INFRARED MICROSPECTROSCOPY
John A. Reffner, Ph.D., Spectra-Tech, Inc. 2 Research Drive, Shelton, CT 06484 --------------------- Members and Non-Members Welcome - Location and Reservation Information at end of this note. Reservation deadline Friday at 12:00.
Newsletter is also available at: http://www.msa.microscopy.com/~psmlas/newsltrs.html ------------------
Abstract
Infrared microspectrometry (IMS) is the measurement of infrared spectra through a microscope coupled to Fourier transform spectrometer. This FT-IR microscope is a tool used to visually detect microscopically small samples or domains and to record their infrared spectra. This instrumentation unites two sciences, microscopy and spectroscopy. IMS has been applied to analysis of adhesives, cosmetics, copy toners, drugs, explosives, fibers, inks, paints, plastics and soils on a truly ultra-microscopic scale. Samples weighing a few nanograms are routinely identified and quantified. When microscopy and spectroscopy are used together, scientists have a greater discrimination power. Infrared spectroscopy resolves chemistry, while microscopy resolves shape and form. IMS is used identify a single fiber or to differentiate a nylon-6 fiber from nylon-6,6 fiber. Today that is not too impressive but, using IMS to separate single acrylic fibers into 23 unique compositional classes has been a major advance for identification of trace evidence in forensic investigations.
IMS places new demands on the analyst --- you must be both microscopist and spectroscopist. Microscopy has a long and distinguished history of measuring, evaluating and comparing materials. Microscopes lets you see more detail, detect unseen structure and solve problems. Add the ability to get infrared spectra of any microscopic domain and your power expands to resolve molecular chemistry.
Microscopy can be defined as the art and science of creating, recording and interpreting magnified images. The combination of SEM with EDX gave us the ability to do elemental analysis, now the molecular chemistry can be probed using the combination of light microscopy with infrared spectroscopy.
------- Sponsored by: KEVEX manufactures X-ray Analyzers, capable of analyzing sample sizes ranging from microns to millimeters, thickness from angstroms to microns, and elemental concentrations from PPM to weight percents.
--------------------------------------------------- NOVEMBER MEETING DETAILS
Wednesday, November 12, 1997 Location: LRSM Building (Laboratory for Research on the Structure of Matter), UPENN, 33rd and Walnut Street (map enclosed). Parking is available behind the LRSM after 5:00 PM. (We have made arrangements with UPENN, they say they will not tow) Cost of Dinner: Members $12.00, Students $6, Non-Members $15. Schedule: 5:30 Social hour. Hosted by our meeting sponsor. 6:30 Dinner Menu: Steamed Dumplings with Plum Sauce, Fried Wontons Chicken Stir Fry, Vegetarian Stir Fry, Rice Spinach Salad with Fresh Mushrooms and Almonds Fruit, Fortune Cookies, Coffee, Decafe, Tea Reservations: By E-Mail (preferred): Send your name and affiliation to PSM-RESERVATIONS-at-INAME.COM By Phone: Call Ms. Pat Overend at (215) 898-8337. DEADLINE for RESERVATIONS is 5:00 PM Thursday November 6th. Reservations are required. We cannot guarantee you a meal if you do not make a reservation prior to the deadline.
About the Speaker
For the past ten years Dr. Reffner has been employed by Spectra-Tech, first as a Corporate Fellow and for the last three years as Research Director. In this role Dr. Reffner has led the technical development of infrared microspectroscopy. Prior to joining Spectra-Tech, he was a Principal Scientist with American Cyanamide (1977 - 87), Assistant Director of the Institute of Material Sciences at the University of Connecticut (1966 - 77) and Research Director at McCrone Associates, Chicago, Illinois (1958 - 66). His undergraduate education was at Akron University. He received a master's degree at Illinois Institute of Technology and a doctorate from the University of Connecticut. In addition to infrared microspectroscopy include polymer science, microscopy and forensic science. He is a Special Consultant to the Connecticut State Police and is on the editorial board of the Journal of Forensic Science.
-- "Opinions expressed are mine and not those of Rohm and Haas Company"
I need some advice. What would be a good journal to publish a technical advance type of paper which reports a methodological improvement for immunoEM studies of plant tissues ?
Thanks in advance,
Soumitra
Soumitra Ghoshroy Department of Biochemistry and Cell Biology State University of New York at Stony Brook Stony Brook, NY 11794-5215 Tel: 516-632-9536 Fax: 516-632-8575
Do you fix your membranes before negative staining?
We do negative stain our fixed samples, and we get beautiful results, and yes, it is easy for us to see the lamination.
Though we've seen nice results from unfixed samples as well.
We use Amonium Molybdate, at pH 6.3
Lou Ann
} [Marcel Paques:] } It is well documented in literature that negative staining deteriorates the } original structure of liposomes. The technique should not be used for their } observation!!!!!!.
Lou Ann Miller Center for Microscopy & Imaging College of Veterinary Medicine Dept. of Veterinary Biosciences University of Illinois Rm 1108 Basic Sciences Bld 2001 S Lincoln Ave. Urbana, Illinois 61802
For the purposes of equipment replacement, costing for microscopy, as well as other reasons our Admin people have asked me to tell them what the life expectancy of electron microscopes is. We need to know this value because it is an integral part of any calculations we have to do with respect to our microscopes.
I have worked on both TEMs and SEMs that have been close to 25 years old, and still running well enough to allow us to get the information we required. I have also talked with some colleagues who have given 10 years as the expectancy, when the equipment, although it still may be working well, is considered to be "outdated".
In this vein, I would be interested to hear from colleagues and commercial folks alike to see what the general consensus is, and what a reasonable value for life expectancy is. This is important because this is the value I will always use in all my calculations from now on.
Thank you.
Paula Allan-Wojtas Food Microstructure Specialist Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
How to tell if liposomes are uni or multi-lamellar ?
Freeze fracture.
Regards
John John Manston Electron Microscope Unit Faculty of Medical Sciences Queen Mary and Westfield College University of London Mile End Road London E1 4NS Tel +171 982 6961 Fax +181 983 0613
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Marcel
Yes I realize that. And we do do freeze fracture and cryo-TEM to see lamellar structures in liposomes. However, we negatively stain liposomal preps prior to freezing to perform preliminary assessment of their quality and density. Often in such collapsed vesicles one gets to see muti-lamellar striations.
It is well documented in literature that negative staining deteriorates the original structure of liposomes. The technique should not be used for their observation!!!!!!.
I always perform the LT observation in TEM of vitrified thin films in parallel with freeze fracture. Discrimination between uni- and multi-lamellar in thin films is straight forward. The freeze fracture in parallel is to have a confirmation of your findings in thin film. The thin film preparation can introduce various artefacts, e.g.: 1 selection of diameter size ( Determined by the film thickness the liposomes are ordered in their diameter. Diameters larger than the thickness of the film
are excluded from the thin film and will not be visible in the TEM. 2.in case of viscous liquids, mechanical stresses are applied during thin film preparation, which can cause phase inversion. 3.Diameter sizes larger than the film thickness present in the film can be deformed and have lost their spherical shape (have become flattened between both air/water interfaces of the film. Tilting of the grid in the beam is needed to check on the shape of the vesicles. In the freeze fracture replica all sizes are present in their actual shape and distribution in the dispersion. Replication is limited in discrimination between uni- and multi-lamellar vesicles. Insight in this aspect is only obtained in case of cross-fractures through vesicles.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi Manoj.
Good question: Not readily, I'm afraid. It is actually difficult to assess whether liposomes are uni- or multi-lamellar with neg staining. Actually, the best method to answer this is cryo-EM of a frozen hydrated suspension of the liposomes.
Regards, Michel
At 18:21 4/11/97 +0100, you wrote: } } Michel, } } I wonder how readily do you get to see if your liposomes are } multilamellar or not? } } Manoj } } Dr. Manoj MISRA, } Unilever Research } 45 River Road } Edgewater, NJ 07020 } (201)840-2702 (voice) } (201)840-8299 (fax) } Manoj.Misra-at-unilever.com } **************************************************** Michel Deschuyteneer, Ph.D. deschuyt-at-sbbio.be Scientist Electron Microscopy Laboratory
SmithKline Beecham Biologicals Rue de l'Institut, 89 B1330 Rixensart, BELGIUM Tel: +32-2-656 9290 Fax: +32-2-656 8164 **************************************************** Standard disclaimer: the opinions expressed in this communication are my own and do not necessarily reflect those of SmithKline Beecham. ****************************************************
Microscopy-at-sparc5.microscopy.com Original-Encoded-Information-Types: IA5-Text X400-Content-Type: P2-1984 Message-ID: {"ISOPRO::DH-EF::1BEA::3460A54A"*/G=Allen/S=White/O=txmta1/PRMD=AMD/ADMD=ATTMAIL/C=US-at-MHS} Importance: normal
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
It's been 25 years since I've been in a micropaleontology prep lab but I do remember that rads are quite durable. They would be soaked them for a couple of days in Quaternary-O, I think a petroleum based dispersant. The clay was dispersed in a blender and then the rads filtered out with a 300 or so mesh sieve. I have seen rads removed from well indurated rocks bordering on chert by an overnight soaking in dilute HF followed by siving.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi:
A user in our lab wishes to image radiolarians (nearly pure SiO2) and separate them from the matrix (smectite to illite clay) in order to do an analysis of area/volume ratios.
In fractured samples we can see the radiolarians easily in the SEM, but he wants to be able to do image analysis on a picture and pick out the rads by doing some thresholding. In our pictures, the rads and the matrix are all similar in gray value.
Whole rads are 60 - 80 um in diameter, but they are also found as 'chunks' as small as 3 um. The clay particles of the matrix are from 1 - 5 um in size. He wants to separate even small particles of rads to get a measure of displacement and volume changes. I suggested that he may have to prepare a polished surface to examine, but he would like to avoid doing that since the samples are somewhat friable and he is not sure how to proceed.
We tried SEI and BSE imaging in the SEM, but the gray values are too close to do much thresholding. We tried doing some x-ray mapping, but Si is abundant in both the rads and the matrix so not much to go on there. Any elements unique to either the rads or the matrix seem to be present in concentrations too low to make a good separation using x-ray maps.
I have told him that I think this is a hard problem, but that I would send a message to the list to see if anyone had any ideas or leads on techniques. So, what do you think?
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
Message-Id: {199711051510.JAA24950-at-mailhub.iastate.edu} X-Sender: wes-at-pop.ameslab.gov X-Mailer: Windows Eudora Pro Version 2.1.2 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
What about using the Al in the clay to note the difference? There should be enough.
Certainly doing x-ray maps is not as fast as SEI or BSE imaging, but it can work. We have done it routinely to pick out phases in cement paste for image analysis.
At 04:01 PM 11/4/97 -0800, you wrote: } Hi: } } A user in our lab wishes to image radiolarians (nearly pure SiO2) and } separate them from the matrix (smectite to illite clay) in order to do an } analysis of area/volume ratios. } } In fractured samples we can see the radiolarians easily in the SEM, but he } wants to be able to do image analysis on a picture and pick out the rads by } doing some thresholding. In our pictures, the rads and the matrix are all } similar in gray value. } } Whole rads are 60 - 80 um in diameter, but they are also found as 'chunks' } as small as 3 um. The clay particles of the matrix are from 1 - 5 um in } size. He wants to separate even small particles of rads to get a measure of } displacement and volume changes. I suggested that he may have to prepare a } polished surface to examine, but he would like to avoid doing that since } the samples are somewhat friable and he is not sure how to proceed. } } We tried SEI and BSE imaging in the SEM, but the gray values are too close } to do much thresholding. We tried doing some x-ray mapping, but Si is } abundant in both the rads and the matrix so not much to go on there. Any } elements unique to either the rads or the matrix seem to be present in } concentrations too low to make a good separation using x-ray maps. } } I have told him that I think this is a hard problem, but that I would send } a message to the list to see if anyone had any ideas or leads on } techniques. So, what do you think? } } Jonathan Krupp ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-8216
Here are my two cents, I would say that: as it's so easy to do cryo EM on liposomes you should never try Neg. Staining. You will just loose your time... And cryoEM give you much more results about the structure of your suspension.
A cryo EMist
------------------------------ SCHMUTZ Marc IGBMC 1 rue Laurent FRIES BP 163 F 67404 Illkirch Cedex FRANCE
Good start. As an iteration on your basic recipe, add more viscosity by using 4 vol% perchloric in equal parts ethanol/methanol/butanol. This can improve the polish, but not always....
David B. Snow Pratt & Whitney Materials & Mechanics Engineering 400 Main St. (MS 114-45) East Hartford, CT 06108 860 565 7823 snowdb-at-pweh.com
Consider the potential dust problem with any but an evaporative humidifer (or used distilled water). Misting units will cause dissolved minerals in the water to precipitate and mineral "snow" will eventually cover everything. Could be a problem in an EM lab.
Woody White, Electron Microscopist SEM/EDS/WDS
Work: Mcdermott Technology, Inc. woody.n.white-at-mcdermott.com http://www.mtiresearch.com/
I'm thinking of installing a humidifier in the lab here, at least for winter use. Has anyone out there been able to successfully regulate a comfortable humidity level in a cool-climate lab? I'm going to have to be careful about approaching management to pay for a humidifier, though, because our building is just now going through some major renovation to take care of a minor outbreak of Stachybotras (a damp-loving mold that can be quite toxic to some people) that occurred in another, much moister part of the complex. So when they hear anything about me wanting to increase a humidity level somewhere, they may get a little squirrely on me.
A fellow student working in our lab is looking for protocols for embedding liposomes in resin. Anyone familiar with protocols other than cryoEM that will work?
Lisa D. Brown University of Connecticut Physiology and Neurobiology Department Electron Microscopy Laboratory Box U-131, Rm 129 Beach Hall Storrs, Ct 06269-2131 Tel. (860)486-2914 Fax. (860)486-1936
We may need a microtome periodically (rm. temp. for now, cryo in the future) while we are performing experiments at Brookhaven National Lab, Upton, NY. We may be in need of the microtome as early as Monday, November 10th. We would have our own supplies, including knives, and our own operator with 15 years microtomy experience. It would need to be within approximately a one hour drive of Brookhaven, readily available and rentable by the hour. If anyone has suggestions please contact me.
Michael T. Dineen The Dow Chemical Company 1897 Building Midland MI 48667 (517/636-4008 4 517/638-6443 + mtdineen-at-dow.com
National Institutes of Health Bethesda, Maryland Laboratory of Structural Biology Research National Institute of Arthritis, Musculoskeletal & Skin Diseases
Technical specialist in support of high resolution macromolecular electron microscopy program (P.I. Dr. Alasdair C. Steven).
Responsibilities involve maintaining/operating/ testing/development of instrumentation (transmission electron microscopes, cryo-holders, freeze-etch machine and cryo-microtome) Experience in and aptitude for these activities is desirable, although some on-the-job training is possible. Also includes responsibilities for management of EM facility. Background in physical or life sciences or bioengineering at BS or MS level plus practical experience. Appointment at GS9 - GS11 level (approx. $31, 680 - $49,831), according to experience.
For further information and detailed instructions about application procedure, please contact :
Ms. Kathy Phelan Bldg. 45, Rm. 5AS53 National Institutes of Health Bethesda, MD 20892 Tel: (301) 435-5315 Fax : (301) 435-5319
wise-at-vaxa.cis.uwosh.edu wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } I'm considering buying some rebuilt SEM filaments (W). I am seeking input } from users as to how they compare to new (manufacturers or second source). } } TIA } } Bob } } Dr. Robert R. Wise } Department of Biology } University of Wisconsin-Oshkosh } Oshkosh, WI 54901 } } (920) 424-3404 tel } (920) 424-1101 fax } wise-at-uwosh.edu
Dr. Wise,
Since Ladd Research has been providing both new and rebuilt tungsten filaments for many years we feel we have no bias either way in this matter. For rebuilt filaments we clean the bases, afix the same configyration of tungsten loop as the EM manufacturer and anneal the filament. The life and preformance should be the same for new and rebuilt. We would suggest rebuilt filaments unless the base is contaminated and can not be cleaned. Some of the older scopes use porous ceramic bases which are differcult to clean and should be replaced.
John Arnott Chairman Ladd Research 13 Dorset Lane Williston, VT 05495 tel 1-800-451-3406 fax 1-802-878-8074
I Have sections of fluorescently labelled rat spinal cord and would like to counterstain the entire section to show a general morphological outline that accentuates white/grey matter, etc. The stain can be either fluorescent, or, more likely, visualized with a light microscope. I am having difficulty finding a genral histology stain which is not autofluorescent.
Any suggestions?
Judy Trogadis Eye Research Institute and University of Toronto Toronto Hospital, Western Div. 399 Bathurst St. Toronto, Canada M5T 2S8
Hi Soumitra, You might consider sending your Technical Advance paper to the Journal of Microscopy Research and Technique.
John E. Johnson is the editor and the editorial office address is: 165 Cervantes Road Redwood City, CA 94062 ph: 415-366-1644 FAX: 415-367-9630
cheers ed
Edward J. Basgall, PhD The Pennsylvania State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618 http://www.personal.psu.edu/ejb11/ Privilege does not absolve one of ecological responsibility.
I have a CL detector attached to my ISI SEM. The detector is not a mirror-type detector. It consists of an acrylic light pipe and a photomultiplier tube. Lately the CL image signals are getting weaker and noisy. I am thinking that the light pipe may get contaminated. How can I clean the detector ? Thanks.
In Posteket al., as well as other SEM texts, it is stated that characteristic x-rays are emitted from a deeper zone in the interaction volume than backscattered electrons. If the depth at which signals are emitted is a function of the energy of that signal, why are the lower energy X-rays (which have energies of ~1kV to ~20 kV at an accelerting voltage of 25 kV) coming from a deeper depth than BSE (which would have energies of ~25 kV)?
Bob
Dr. Robert R. Wise Department of Biology University of Wisconsin-Oshkosh Oshkosh, WI 54901
(920) 424-3404 tel (920) 424-1101 fax wise-at-uwosh.edu
If you are using FITC, I have had good success couterstaining with .01% Evans Blue. It is fluorescent with texas red filter set and works great as a total counterstain with tissue labelled with FITC.
Bob
On Wed, 5 Nov 1997, Judy Trogadis wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I Have sections of fluorescently labelled rat spinal cord and would like to } counterstain the entire section to show a general morphological outline } that accentuates white/grey matter, etc. The stain can be either fluorescent, } or, more likely, visualized with a light microscope. I am having difficulty } finding a genral histology stain which is not autofluorescent. } } Any suggestions? } } Judy Trogadis } Eye Research Institute and } University of Toronto } Toronto Hospital, Western Div. } 399 Bathurst St. } Toronto, Canada M5T 2S8 } } phone: 416-603-5088 } Fax: 416-603-5126 } email: judy-at-playfair.utoronto.ca } } }
Bob Wise wrote: } } I'm considering buying some rebuilt SEM filaments (W). I am seeking input } from users as to how they compare to new (manufacturers or second source). } ------------------------------------------------------------------------------- } -----------------------------
I've bought reconditioned filaments for several years and I have not noticed any differences compared to new filaments. Occasionally I have bought new filaments from a "second source".
Russell E. Cook Scientific Associate Electron Microscopy Center for Materials Research Argonne National Laboratory Building 212 9700 South Cass Avenue Argonne, IL 60439 (630)252-7194 FAX: (630)252-4798
Anybody out there know how to contact a Balzar's rep in the US. I am interested in getting info on their high pressure freezer.
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
Kindly instruct me on how to be removed from this list. I have tried sending unsubscribe messages 3 times to ListServer-at-MSA.Microscopy.Com but I am still receiving mail from this list. Thank you.
------------------ GINNY E. CRUZ Section of Immunopathogenesis, Institute of Immunological Science Hokkaido University : Kita Ku Kita-15 Nishi-7 Sapporo City 001 JAPAN Tel. No.: (011)716-2111 Ext. 5120 Fax No.: (011)736-9836 e-mail: gcruz-at-imm.hokudai.ac.jp ------------------
This is a good question, but you are not the first person asking it.
I think that the answer is simple. You have only considered the absorption of the sample to the signals rather than the primary electron beam.
We know that the BSE signal is produced by the interaction between the primary electrons and the atomic nuclei of the sample, while the X-ray signal is the side product of the interaction between the primary electron beam and the outer layer electrons of the atoms in the sample. As the depth increased, the primary electrons would loss some energy, they would loss the qualification to produce the high energy BSE at some stage, but they can still create the X-ray in a deeper or/and wider zone. Then, yes, you are right, we should also consider the absorption of the sample to the signals.
Hope this can help.
Regards,
Charlie Kong
wise-at-vaxa.cis.uwosh.edu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } In Posteket al., as well as other SEM texts, it is stated that } characteristic x-rays are emitted from a deeper zone in the interaction } volume than backscattered electrons. If the depth at which signals are } emitted is a function of the energy of that signal, why are the lower } energy X-rays (which have energies of ~1kV to ~20 kV at an accelerting } voltage of 25 kV) coming from a deeper depth than BSE (which would have } energies of ~25 kV)? } } Bob } } Dr. Robert R. Wise } Department of Biology } University of Wisconsin-Oshkosh } Oshkosh, WI 54901 } } (920) 424-3404 tel } (920) 424-1101 fax } wise-at-uwosh.edu
Dear Bob, That's easy, Bob. X-rays travel further through solid material than electrons, probably because they are heavier ;-) You wrote: } In Posteket al., as well as other SEM texts, it is stated that } characteristic x-rays are emitted from a deeper zone in the interaction } volume than backscattered electrons. If the depth at which signals are } emitted is a function of the energy of that signal, why are the lower } energy X-rays (which have energies of ~1kV to ~20 kV at an accelerting } voltage of 25 kV) coming from a deeper depth than BSE (which would have } energies of ~25 kV)? } } Bob } } Dr. Robert R. Wise } Department of Biology } University of Wisconsin-Oshkosh } Oshkosh, WI 54901 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 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
I wonder if anybody can help a colleague of mine? She is in need of= :
"a conical molybdenum insert cap for a Wehnelt asembly used in a Philips 500 series SEM, when fitted with a low kV anode"
I don't understand the question, so please can any replies be addressed directly to:
Ann.Hayes-at-bbsrc.ac.uk
Many thanks in advance!
Nigel Chaffey
----------------------------------------------------- Dr Nigel Chaffey, Dept Forest Genetics & Plant Physiology, Swedish University of Agricultural Sciences, S-901 83 Ume=E5, Sweden Phone: +46-90-786-6305 =46ax: +46-90-786-5901 eMail: nigel.chaffey-at-genfys.slu.se
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Paula,
The opinion of my company is a life expectancy of 10 years. The depreciation is calculated based on that period and the microscope will be replaced after 10 years.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello, everyone,
For the purposes of equipment replacement, costing for microscopy, as well as other reasons our Admin people have asked me to tell them what the life expectancy of electron microscopes is. We need to know this value because it is an integral part of any calculations we have to do with respect to our microscopes.
I have worked on both TEMs and SEMs that have been close to 25 years old, and still running well enough to allow us to get the information we required. I have also talked with some colleagues who have given 10 years as the expectancy, when the equipment, although it still may be working well, is considered to be "outdated".
In this vein, I would be interested to hear from colleagues and commercial folks alike to see what the general consensus is, and what a reasonable value for life expectancy is. This is important because this is the value I will always use in all my calculations from now on.
Thank you.
Paula Allan-Wojtas Food Microstructure Specialist Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
} } I need to prepare TEM specimens of Inconel 718 superalloy . I was going to start with perchloric and ethanol at about - 35 C. I would however appreciate getting "recipes" from people with more experience on this matter. { {
Hi Jordi,
we routinely prepare TEM specimen from superalloys, both single crystal and poly crystalline (e.g. SC16, IN738 etc.) for our study of deformation substructure. We use twin jet polishing technique under following condition and get good sucess:
10% perchloric acid + 90% ethanol at 263 K and 23 V
It should work equally good for IN718. Wish you sucess.
-- Dr. D. Mukherji Group NM, Hahn Meitner Institut Glienicker Strasse 100 D-14109 Berlin, Germany Tel. (030) 8062 3099 Fax. (030) 8062 3059
The best journal for your paper if it is good science is, of course the Journal of Microscopy. Contact Sue Betteridge at jmicrosc-at-rms.org.uk for details.
Patrick Echlin General Editor Journal of MicroscopyOn Wed, 5 Nov 1997, Soumitra Ghoshroy wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hi Fellow Microscopists, } } I need some advice. What would be a good journal to publish a technical } advance type of paper which reports a methodological improvement for } immunoEM studies of plant tissues ? } } Thanks in advance, } } Soumitra } } Soumitra Ghoshroy } Department of Biochemistry and Cell Biology } State University of New York at Stony Brook } Stony Brook, NY 11794-5215 } Tel: 516-632-9536 } Fax: 516-632-8575 } } }
I need some advice. I was asked to do some analysis of silica particles (size distribution) for chemist in our institute. Particle size should be in the range of 3 to 6 um. I do not have any experiences with such sample. Could someone give me a tip how to prepare sample for TEM (or SEM)?
Thanks in advance, O. Benada
+---------------------------------------------------------------+ Oldrich Benada Acad. Sci. CR Phone: +420-2-4752399 Institute of Microbiology Fax: +420-2-4715743 Electron Microscopy Group E-mail: benada-at-biomed.cas.cz Videnska 1083 CZ - 142 20 Prague 4 - Krc Czech Republic +---------------------------------------------------------------+
There are few ways that I have had sucess at preparing paper cross-sections. The most commonly use method involves embedding in a low viscosity resin (Spurr's) and facing the cross-section with an diamond ultramicrotomy knife. The alternative method is to prepare the sample similary to a metallurgical specimen. Again using a low viscosity resin rather than standard epoxies.
The problem with epoxies will arrise with the FT-IR analysis. I have nerver tried this method, but you might infiltate sample with water and try cryomicrotomy. This will work as long as you do not mind swelling of the paper.
If you need more information, contact me at john_catino-at-ucamp.com
3D reconstruction from TEM images requires high resolution digital images. With the use of scanners with spot sizes of down to 7 microns a single sheet of em film produces a huge image file. With the need to collect hundreds/thousands of such images to improve resoultion/noise in reconstructs what are peole doing to store all these files? The files need to be stored so that they can be retrieved sensibly so tape archiving is not suitable. Is the technology available and at what cost?
Looking forward to a stimulating discussion.
Chris
Chris Gilpin Biological Sciences Electron Microscope Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 161 275 5170 fax +44 161 275 5171 http://www.biomed.man.ac.uk/biology/emunit/emhome.html
From all sources which I have read indicate that all else being equal, x-rays are more penetrating than electrons. That is to say that for electrons and x-rays of equal energy electrons are more easily shielded. "Hair splitter": Emission of x-rays and generation of BSEs can occur anywhere in the volume, only the more penetrating x-rays make it to the surface from a larger volume.
Woody White, Electron Microscopist SEM/EDS/WDS
Work: Mcdermott Technology, Inc. woody.n.white-at-mcdermott.com http://www.mtiresearch.com/
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
In Posteket al., as well as other SEM texts, it is stated that characteristic x-rays are emitted from a deeper zone in the interaction volume than backscattered electrons. If the depth at which signals are emitted is a function of the energy of that signal, why are the lower energy X-rays (which have energies of ~1kV to ~20 kV at an accelerting voltage of 25 kV) coming from a deeper depth than BSE (which would have energies of ~25 kV)?
Bob
Dr. Robert R. Wise Department of Biology University of Wisconsin-Oshkosh Oshkosh, WI 54901
(920) 424-3404 tel (920) 424-1101 fax wise-at-uwosh.edu
Absorbance of X-rays is lower then absorbance of electrons.
wise-at-vaxa.cis.uwosh.edu wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } In Posteket al., as well as other SEM texts, it is stated that } characteristic x-rays are emitted from a deeper zone in the interaction } volume than backscattered electrons. If the depth at which signals are } emitted is a function of the energy of that signal, why are the lower } energy X-rays (which have energies of ~1kV to ~20 kV at an accelerting } voltage of 25 kV) coming from a deeper depth than BSE (which would have } energies of ~25 kV)? } } Bob } } Dr. Robert R. Wise } Department of Biology } University of Wisconsin-Oshkosh } Oshkosh, WI 54901 } } (920) 424-3404 tel } (920) 424-1101 fax } wise-at-uwosh.edu
We have an immediate opening for an electron microscopist to perform microstructure studies on magnetic materials, including small particles, thin films/multiilayers, and permanent magnets. The Electron Microscopy Lab (Physics and Astronomy Dept., University of Delaware) consists of a Jeol JEM 2000 FX, a Jeol JEM 100 CX, and an Amray 1200 SEM.
The position is for one year initially, and can be renewed for the next three years.
Interested parties please respond by sending a resume and names and contact information for three references:
(a) by FAX to George Hadjipanayis at 302-831-1637 or
(b) by e-mail to: pmusa-at-udel.edu (Only in ASCII text or as part of a regular e-mail message, please.)
The University of Delaware is an Equal Opportunity Employer.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Patricia Meadows e-mail: pmusa-at-udel.edu Physics & Astronomy Dept. Phone: 302-831-2662 University of Delaware FAX: 302-831-1637 Newark, DE 19716-2570, USA ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Another fine microscopy journal, if it is good science, is MICROSCOPY AND MICROANALYSIS, the official journal of the Microscopy Society of America, the Microbeam Analysis Society, the Canadian Microscopical Society, and the Mexican Microscopy Society. It is published by Springer-Verlag.
For information you can consult the journal's web site:
A. Kent Christensen Department of Anatomy and Cell Biology Medical Sciences II Building University of Michigan Medical School Ann Arbor, MI 48109-0616 akc-at-umich.edu Tel (313) 763-1287 http://www.umich.edu/~akc/
-----------------------------------------
} } } } Hi Fellow Microscopists, } } } } I need some advice. What would be a good journal to publish a technical } } advance type of paper which reports a methodological improvement for } } immunoEM studies of plant tissues ? } } } } Thanks in advance, } } } } Soumitra } } } } Soumitra Ghoshroy } } Department of Biochemistry and Cell Biology } } State University of New York at Stony Brook } } Stony Brook, NY 11794-5215 } } Tel: 516-632-9536 } } Fax: 516-632-8575 } } } } } } } }
Sorry for being a little slow, but I see no one else has responded to the list regarding your inquiry.
With regards to your question about "loosinging" the cells that's easy. I've worked with flask grown cultured cells a number of times, the easiest method for dealing with surface attached cells (generally in the retangular, lay-down 'plastic' culture flasks) is to:
(1) dump out the media
(2) add the fixative directly into the flask (make sure you use enough to cover the cell layer)
(3) Wait for the recommended primary fix length (hopefully someone else will provide information on appropriate fix and times) and dump out fix.
(4) rinse with appropriate rinse agent (i.e. buffer? unless ddH2O is preferred).
(5) Dump out first rinse, add second (to reduce residual fixative remaing, and make things safer) With a flask scraper (ask the dermatologist providing the sample for one - they look like tiny rubber window washer squeegies attached with a small hinge on the end of a plastic handle, they use these for transferring cells from one flask to another, and are designed to gentlely remove cells adherent to flasks) gentlely scrap the cells from the surface of the flask. Following fixation I have found that confluent cell layers adhear toe ach other very nicely and come off as strips and small sheets (i.e. 1-3mm x 1-3 mm x 1-3 cells deep). With extra buffer rinse these cell 'strips' into a test-tube or vial.
(6) Allow the cells chunks to settle, or breifly centrifuge LIGHTLY. And remove most of the excess rinse buffer. This should give you a concentrated suspension to work with.
(7) Mix the concentrated cell suspension with ~ equal amount of COOL (~ 45 -50 C) 2-4% agar/agarose in H2O. NOTE: Agar/agarose must be heated to ~100 C to get it into suspension but then doesn't gel until ~41C. I have found a 45C water bath ideal for keeping agar/agarose molten and at the right temperature. BUT it will set up very rapidly if you add cold cell suspension to 45 C agar/agarose. Since the cells are grown at 37C any way warming the fixed cells to 45C with the Agar/agarose should not cause problems.
(7) You can use epindorf tubes to mold the cell/agar mix or mix rapidly and poor out into a clean/sterile petri dish. Dice up either solidified mixture with a razor blade, and treat the agar/cell blocks just like any other tissue. If you osmicate, the agar will not turn black but the cells will (making them easier to find!). Remember that the 'agar' will be ~1-2% stuff and 98-99% empty space so infiltration into the agar is not much of a problem.
(8) The agar doesn't pick up much EM staining (little more LM staining) and exhibits only slightly greater electron density than empty resin.
Good luck.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"640K ought to be enough for anybody." -- Bill Gates, 1981
Would anyone know details of suppliers where Uranyl acetate can be obtained already in solution i.e. a saturated solution in 50% ethanol. We previously made this up ourselves from dry powder and ethanol, but we are trying to reduce the potential hazzard to health by purchasing ready made solution. Thanks on advance Orla O'Shea, Dept of Anatomy, QUB o.oshea-at-qub.ac.uk
I was just reading a section in Russ's book (Handbook of Image Processing, CRC Press) on using some gray-processing kernels that are sensitive to texture. I haven't tried this, and have no idea about software, but the examples in the book make it seem worth a try.
TEM - Input required for procedure to stain poly(4-vinyl pyridine).
I am interested in observing domain size in a block copolymer film of composition; poly(styrene-b-4-vinyl pyridine). So far I have made an attempt using OsO4 without any success. I could give RuO4 a try but I understand that it will likely stain the polystyrene as well. Any suggestions?
Thanks to all of you who responded to my inquiry about where to publish a technical advance paper. It was a great help.
Soumitra
Soumitra Ghoshroy Department of Biochemistry and Cell Biology State University of New York at Stony Brook Stony Brook, NY 11794-5215 Tel: 516-632-9536 Fax: 516-632-8575
In message {2.2.32.19971106062706.008f35bc-at-pop.unixg.ubc.ca} , Mary Mager {mager-at-unixg.ubc.ca} writes } That's easy, Bob. X-rays travel further through solid material than } electrons, probably because they are heavier ;-)
Photons are only heavier in reciprocal space of course. Outside of the electron microscope they will revert to their normal mass :-)
The mechanism of absorbtion is somewhat different. Electrons are simply scattered to lower and lower energy. You get a typical 1/E2 billiard ball kinematics curve for the absorbtion. But photons can be absorbed as quanta so the absorbtion curve for x_rays is a complex shape with distinct 'edges'.
The matter of incident beam path is an additional factor, significant but not primary (I think).
-- Usual disclaimers. Anthony James Bentley Surface Data Scientific Instrumentation and Software Web site http:\\www.surface.demon.co.uk
The generation of X-rays in terms of depth in the interaction volume is determined by several factors. X-rays will be produced throughout the volume until the energy of the x-ray is below the ionization energy of the core level responsible for that x-ray. Characteristic x-rays from different elements are generated from different volumes having different depths and the depths are strongly dependent on composition of the sample. In general, the depths of these volumes are deeper for lower edge energies and are deeper for lower average atomic number samples. The x-rays have a longer mean free path in a material than an electron; the measure of this is in the value of the cross section for scatterring. The absorption of an x-ray is an all or nothing proposition, once it is absorbed, it is gone. However, the electron can lose some of its energy during a scatterring event. The backscatterring process is occurring deeper in the sample, but the electron is loosing its energy on the way out of the sample. All of the electrons having energies from 50 eV (by definition) up to the primary are essentially backscattered electrons. There is a peak in the distribution near the primary energy. The electrons in this peak are coming from the near surface region because they haven't lost much energy. Incidently, this is why Auger electron spectroscopy is so suface dependent. The Auger electron generation process is also occurring with in the electron interaction volume below the surface, but they loose their characteristic energy on their trek to escape from the sample surface. They just get lost in the middle of the backscattered electron distribution and contribute to the background in the Auger spectrum.
In terms of imaging, a backscattered image will always have a poorer spatial resolution for a bulk sample than the secondary electron image. The lower the atomic number of the sample, the poorer the backscattered resolution will be for a given energy. If you lower the beam energy, you will improve the backscattered resolution, but you will be sacrificing detection efficiency of the backscattered elctrons. An x-ray map image will have poorer spatial resolution than a backscattered image.
Hope this helps.
-Scott
Scott D. Walck PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of S.D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
In Posteket al., as well as other SEM texts, it is stated that characteristic x-rays are emitted from a deeper zone in the interaction volume than backscattered electrons. If the depth at which signals are emitted is a function of the energy of that signal, why are the lower energy X-rays (which have energies of ~1kV to ~20 kV at an accelerting voltage of 25 kV) coming from a deeper depth than BSE (which would have energies of ~25 kV)?
Bob
Dr. Robert R. Wise Department of Biology University of Wisconsin-Oshkosh Oshkosh, WI 54901
(920) 424-3404 tel (920) 424-1101 fax wise-at-uwosh.edu
This is more visual optics than microscopy, but can anyone direct me to a good explanation of the visual phenomenon called Haydinger's Cross? This refers to the ability of the human eye to visualize a maltese cross pattern when looking at a white field, with a blue bar in one direction and a yellow bar at 90 degrees. It is apparently caused by some kind of dichroism in the eye.
Gary Radice, Associate Professor gradice-at-richmond.edu Department of Biology 804-289-8107 (voice) University of Richmond VA 23173 804-289-8233 (FAX)
You didn't mention the file size, but for large images files I have been using a writeable CD (CD-R). It is anyone's guess how long technology will be around to read the CDs, but seems the best bet at this time. DVDs are a coming possibility for even more storage. The last blank CDs I bought were {$1.50 each. For 650 megs of storage, "that ain't bad".
Woody White, Electron Microscopist SEM/EDS/WDS
Work: Mcdermott Technology, Inc. woody.n.white-at-mcdermott.com http://www.mtiresearch.com/
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear List
3D reconstruction from TEM images requires high resolution digital images. With the use of scanners with spot sizes of down to 7 microns a single sheet of em film produces a huge image file. With the need to collect hundreds/thousands of such images to improve resoultion/noise in reconstructs what are peole doing to store all these files? The files need to be stored so that they can be retrieved sensibly so tape archiving is not suitable. Is the technology available and at what cost?
Looking forward to a stimulating discussion.
Chris
Chris Gilpin Biological Sciences Electron Microscope Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 161 275 5170 fax +44 161 275 5171 http://www.biomed.man.ac.uk/biology/emunit/emhome.html
Message-Id: {199711061738.RAA06198-at-highgate.mluri.sari.ac.uk} Comments: Authenticated sender is {mi596-at-highgate}
Can anybody explain to me exactly how a film thickness monitor for Au and C coating works. I understand it consists of an oscillating crystal -what is it made of? I have heard accuracy is poor but reproducability is good using same conditions (time, current, working distance etc.). Thickness of C is important from the point of view of quantitative analysis and that unknowns must be coated with the same thickness as the reference standards. Is it sufficient to say to NAMAS or ISO accreditors that the same coating conditions were used for the standards as well as unknowns. Many thanks in advance.
Martin J. Roe MLURI Craigiebuckler Aberdeen AB158QH Scotland U.K.
There is a error in the listing of the program LENZPLUS.BAS on page 429 of the second edition of Electron Energy-Loss Spectroscopy in the Electron Microscope (Plenum, 1996): line 135 should contain ^3*, not ^*. However, this is a typographical error and the program listing at the ftp site (ftp.phys.ualberta.ca) has always been correct.
In addition, error-free execution of the Kramers-Kronig program KRAKRO.FOR, as listed on p.415, requires that the array D be dimensioned as D(8192) in the FFT subroutine, not D(4096) as required on p.413. I have recently made this change to the ftp-site listing.
I am not aware of any other significant errors in the Second Edition but would be grateful for any feedback from readers.
Ray Egerton, Physics Dept, University of Alberta, Edmonton, Canada T6G 2J1 Phone: 403-492-5095, FAX: 403-492-0714, e-mail: egerton-at-phys.ualberta.ca ------------------------------------------------------------------------
} I need some advice. I was asked to do some analysis of silica } particles (size distribution) for chemist in our institute. Particle } size should be in the range of 3 to 6 um. I do not have any } experiences with such sample. Could someone give me a tip how to } prepare sample for TEM (or SEM)? } } Thanks in advance,
} Probably the best way to prepare these samples is to dilute them in a solution of distilled water and to disperse them for several minutes in an ultrasonic bath. Pipette a small drop of the suspension on to a small glass slide (mounted on to a SEM stub by means of a carbon adhesive tab) and allow sample to dry. Coat the samples with a thin layer of Au or Au-Pd and examine in the SEM. You could also try dipping an SEM stub (with a carbon adhesive tab stuck on it) directly in to the sample and blowing off the excess with a Dust-off spray and then coat the sample. I suggest you try the latter of these two options first
Re: particle sizing. I don't do EM, but I do particle size measurements by light scattering and other techniques, using LM to confirm measurements qualitatively for micron sized particles. Microscopy looks at very few particles, so if you care about a measurement that will be valid for a much larger quantity, you really have to think hard about getting a representative sample. This is not easy.
A possibly useful reference is "Sampling for Chemical Analysis," B. Kratochvil, D. Wallace, & J. K. Taylor, Anal Chem. vol 56, 113R-129R (1986).
Leonard Corwin Fort Dodge Animal Health Princeton NJ 08543-0400
Does anyone know of a source of glutaraldehyde powder? We would like to try glut in acetone and other solvents for freeze-substitution, but glut comes only in ethanol as far as I can tell. I suppose we could dry it down from 70% solution, or does this alter it chemically??
TIA,
Rosemary White Department of Biological Sciences Monash University, Melbourne, Victoria 3168, Australia phone 61-3-9905 5670 fax 61-3-9905 5613 email r.g.white-at-sci.monash.edu.au
Hello, I just read a reply to your inquiry. I have prepared cultured cells, too. I suggest initially using a rubber policeman pushing with constant pressure..and not lifting anymore than necessary to get sheets of cells off the dish. Use the solution in dish to flush cells to one edge. I then use a large bore pipette to pick up cells and place into a microfuge (Eppendorf) tube, let the cellular material settle for a minute or two. Then take off the supernatant, leaving cells and just enough solution to keep them covered. Now add your fixatice, ten times the cellular volume, gently mixing. I usually used a modified Karnovsky in buffer for our cells, then centrifuge at a low speed, about 500 rpms, for 10 minutes. If you have enough cells to have a pellet then you don't have to add agar. Continue to fix for at least an hour. If your pellet is large, you will need to loosen it carefully so subsequent solutions can get in from top and bottom. If pellet size is small, all processing can be done in the tube, even the embedding. Just don't fill tube to top, for the polymerizing step, with more media than one would put into a BEEM capsule. After polymerizing, the tube can be cut off and the sides can be trimmed flat to fit into a microtome chuck.
I found that if I first fixed the cells, then tried to scrape them, that too many of them were ruptured. It may be that your cells are not so fragile. Anyway, I then would only recommend a very brief fix before scraping. Kaye
(Roberta) Kaye Brabec, Manager E-mail: brabec-at-umich.edu Morphology Core Facility, Box 0616 Phone: (313) 763-0150 4742 Med Sci II, U of MI Fax: (313) 763-1166 Ann Arbor, MI 48109-0616
I am looking to use a 3-CCD RGB camera that will integrate for at least 5 seconds.
I need to use it to image 3-color FISH samples.
I was wondering if anyone out there has any experience with 3-CCD RGB cameras and can give me some information on a camera that will perform well in terms of resolution, image quality, etc.
We use a twin-jet electropolishing unit to prepare TEM foils of superalloy. We tested several polishing solutions employing a variety of conditions. The best results for our alloys (SRR99 and Udimet 720) were obtained in the use of a solution and condition as following:
Solution: 25% Nitric acid in 75% Methanol Current 0.10 mA Voltage 34~36 V Temperature -35 ~ -45 0C
Good Luck!
Peiyi Wang
Research Fellow Dept. of Engineering Materials University of Southampton Southampton SO17 1BJ UK
} } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } I need to prepare TEM specimens of Inconel 718 superalloy . I was } going to start with perchloric and ethanol at about - 35 C. I would however } appreciate getting "recipes" from people with more experience on this } matter. } } Thanks } } Jordi Marti }
Our lab does a lot of work with semiconductors. My concern is with the preparation of InP, InGaAsP, GaAs, GaP, etc. What are the hazards with these materials? What sort of precautions are required when cutting, grinding and polishing the wafers? I'm finding it difficult finding any safety related information regarding the use of these materials in an EM lab. Thanks in advance.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have lots of experience preparing GaP and GaAs TEM samples. One precaution, GaP reacts with water to produce phosphine, a particularly nasty gas. Phosphine is pyrophoric, so don't be surprised if your samples explode on you if you core out 3 mm disks using an ultrasonic disk grinder. It's happened to me on several occasions. Also, do all your sample prep in a fume hood. While the levels of phosphine emitted by the sample are small (so I assume since I haven't killed myself, yet). You'll get a painful headache from small exposures to the gas. For GaAs, I'm unaware of any similar health issues and have had no difficulties working with this material on an open bench top.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Our lab does a lot of work with semiconductors. My concern is with the preparation of InP, InGaAsP, GaAs, GaP, etc. What are the hazards with these materials? What sort of precautions are required when cutting, grinding and polishing the wafers? I'm finding it difficult finding any safety related information regarding the use of these materials in an EM lab. Thanks in advance.
I 've been asked if it's possible to use microscopy to count ultrafine particles (smaller than 0.1 microns) and to measure their size distributions in an automated fashion. The primary goal is to characterize ultrafines in ambient air. Particles larger than 0.1 microns are typically collected on a screen membrane such as polycarbonate. Most of the ultrafines however would pass through the smallest pores in screen-type membranes. My question is really two-fold: 1) Is it even possible to collect ultrafines for microscopy, and 2) Which technique would be best suited to count and size the ultrafines -- SEM, FE-SEM, TEM, AFM? Or should we just forget microscopy and purchase some particle sizing/counting instrumentation? Thanks for your ideas.
South Bay Technology, Inc. is offering a series of TEM Sample Preparation=
Workshops which will be kicked off with a workshop on RF Plasma Cleaning for TEM and SEM applications. This workshop will be held on Monday December 1, 1997 in Boston, MA.
For a detailed workshop description and registration information on the Plasma Cleaning workshop, please contact me off line and I will forward t= he information to you.
Other upcoming workshops include:
Tripod Polishing for TEM March 13-14, 1998 San Clemente, CA Low Energy on Milling March 12, 1998 San Clemente, CA Plasma Cleaning for TEM March 11, 1998 San Clemente, CA MicroCleave Techniques =
for TEM Sample Preparation March 10, 1998 San Clemente, CA
For information on any of these workshops, please contact me.
South Bay Technology, Inc. is offering a series of TEM Sample Preparation=
Workshops which will be kicked off with a workshop on RF Plasma Cleaning for TEM and SEM applications. This workshop will be held on Monday December 1, 1997 in Boston, MA.
For a detailed workshop description and registration information on the Plasma Cleaning workshop, please contact me off line and I will forward t= he information to you.
Other upcoming workshops include:
Tripod Polishing for TEM March 13-14, 1998 San Clemente, CA Low Energy on Milling March 12, 1998 San Clemente, CA Plasma Cleaning for TEM March 11, 1998 San Clemente, CA MicroCleave Techniques =
for TEM Sample Preparation March 10, 1998 San Clemente, CA
For information on any of these workshops, please contact me.
Oldrich Benada wrote: ===================================================== I need some advice. I was asked to do some analysis of silica particles (size distribution) for chemist in our institute. Particle size should be in the range of 3 to 6 um. I do not have any experiences with such sample. Could someone give me a tip how to prepare sample for TEM (or SEM)? ====================================================== The problem is that those pesky silica particles don't know that they are supposed to separate and stay away from each other when dispersed in a liquid followed by a droplet of this liquid suspension being placed on a solid surface. They tend to agglomerate very quickly leading to a difficult- to-analyze situation, especially using automated means of analysis. You are correct in that the size range expected could be on the order of 3-6 nm.
This is the ideal application for the camphor/naphthalene method which I described several years ago. Credit for the technique, or at least the one who taught it to me was an innovative microscopist then working at the DuPont Experimental Station in Wilmington, DE by the name of Robert P. Schatz, in about 1968, now deceased. Take a 60% camphor/40% naphthalene mixture and heat it to twenty or so degrees above room temperature on a hot plate in a small beaker or flask, the two organics are miscible in each other and this is the eutectic composition.
Once a clear liquid, add a small amount of the silica (not more than 0.1%), which disperses quite readily. Then, using a pipette, take out some liquid and put a drop onto a carbon coated glass slide, at which time the drop is instantly frozen solid (it is at room temperature). Put the slide into your vacuum evaporator to pump out all night, and the "magic" is that the solid eutectic sublimes at room temperature at a rate that by morning, it is completely gone, leaving the silica particles uniformly dispersed on the carbon film!
The rest is obvious. You can pick this up on a grid, as is, or in order to bring out more contrast, Pt/C shadow, probably using an angle not more than 30 degrees. You can float the "replica" off of the slide directly onto a grid and viola! you have particles completely dispersed, virtually no doublets or triplets, and a field quite amenable for automated image analysis (as a bonus).
One important further suggestion: Some times these silica particles tend to fuse together as little "chains". If you suspect this is happening, be sure to take the micrographs as stereo pairs because you can in fact capture this three dimensional spatial information.
Disclaimer: We do not sell either the camphor or naphthalene so have no vested interest in whether people use this method or not. It is just a really neat method for the preparation of fine particle samples in this size range. We are obviously set up to use this method as a service for others, however.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Dear Martin, I believe the film thickness monitors are quartz crystal oscillators that have the characteristic of changing their frequency of oscillation as their mass changes. As you evaporate carbon onto the monitor its mass goes up and the frequency of oscillation goes down. I don't know about the requirements of NAMAS or ISO. You wrote: } } Can anybody explain to me exactly how a film thickness } monitor for Au and C coating works. I understand it consists of an } oscillating crystal -what is it made of? I have heard accuracy is } poor but reproducability is good using same conditions (time, } current, working distance etc.). } Thickness of C is important from the point of view of } quantitative analysis and that unknowns must be coated with the } same thickness as the reference standards. Is it sufficient to } say to NAMAS or ISO accreditors that the same coating conditions were } used for the standards as well as unknowns. } Many thanks in advance. } } Martin J. Roe } MLURI } Craigiebuckler } Aberdeen } AB158QH } Scotland 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 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
Dear Andy, When we worked with GaAs the only precautions were to gather all bits for proper disposal and avoid generating or inhaling any dust, ie. grind and polish wet. Also avoid excess heating because of the possibility of driving off As. You wrote:
} Our lab does a lot of work with semiconductors. My concern is with the } preparation of InP, InGaAsP, GaAs, GaP, etc. What are the hazards with } these materials? What sort of precautions are required when cutting, } grinding and polishing the wafers? I'm finding it difficult finding any } safety related information regarding the use of these materials in an EM } lab. } Thanks in advance. } } Andy
} Andy M. Duft } Brockhouse Institute for Materials Research } McMaster University } 1280 Main St. West } Hamilton Ontario } Canada L8S 4M1 } } 905-525-9140, X24609 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 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
Thanks to all who answered my question about interaction volumes, X-rays, and BSEs.
Stephanie Wind McCray (Moltech Corp) probably had the best answer (or at least the one I understood the best), "keep in mind that backscattered electrons are electrons, and x-rays are photons. There are somewhat different rules for their relative travels through matters of varying atomic numbers" There were a lot of other good answers too.
So (he says with a light bulb over his head), this is why hospitals take X-ray photographs of broken bones and not electron-graphs.
Thanks again
Bob
Dr. Robert R. Wise Department of Biology University of Wisconsin-Oshkosh Oshkosh, WI 54901
(920) 424-3404 tel (920) 424-1101 fax wise-at-uwosh.edu
Hello everybody, a question to the mineralogists on the list: as the German provider (Krantz) for storage boxes for mineralogical standard thin sections cannot provide them any longer I am looking for some other source, preferably in Europe. What I call "standard thin sections" are 28 x 48 mm sized. Thank you in advance Hiltrud
Dr. Hiltrud Mueller-Sigmund Institut fuer Mineralogie, Petrologie und Geochemie Albertstrasse 23b, 79104 Freiburg (Germany) Tel.: (+49)-203-6388/-6396 Fax: -6407
"Calibration of an Off-Axis Quartz Crystal Thickness Monitor for a Pulsed Laser Deposition System Using a High Resolution Scanning Electron Microscope", S. D. Walck, J. S. Zabinski, M. S. Donley, and J. E. Bultman, Thin Solid Films, 236, pp. 125-9, 1993.
If you look that paper up, I bleive that there are two excellent references in there on how a thickness monitor works. One of them is a journal article that apparently is a classic and the other is a reference manual for the XTC quartz crystal thickness monitors. These references talk about some of the limitiations on the technique and accuracies etc. I would give you the references myself, but my papers are still boxed up from my move to PPG -sorry. You could contact Jeff Zabinski at Wright Patterson AFB and get a reprint of that paper since they have it on file there. He can be reached vis Email at zabinsj-at-ml.wpafb.af.mil
-Scott Walck
Scott D. Walck PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of S.D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Good Afternoon, I've enjoyed reading the many postings on the Microscopy Listserver. Now I have a question for all of you...
Has anyone noticed a change in the quality of TEM Grids?
For years we've ordered our TEM grids from Biorad.... We've used the Thin bar Hexagonal 460 mesh copper and nickel grids for years. They were always very clean, very consistent in thin bar width, and of excellent quality.
Now that they are no longer selling them, we've been trying grids from several sources. I have done a comparison of grids from different companies. A number of them are close, but not quite as thin as the original Biorad grids. Others have too much variation in grid width (even within the same vial) and some are unacceptable, too wide, more like normal grid bars, and not thin bars....
Does anyone know the original source of the grids that Biorad used to sell? None of the companies we've tried have the quality we are looking for... some are close, but even within the same vial of grids, we've found too much variation in the grid bar width.
Thank you for your assistance,
Virginia Tanner Crocker
******************************************************************* Virginia Tanner Crocker Biologist NIH, NINDS EM Facility, Bldg 36, Room 3B24 Bethesda, MD 20892
There is a description of the phenomenon of "Haidinger's brush" (or cross) in "Condepts of Classical Optics" by John Strong, Freeman, San Francisco, 1958, P.111f. He quotes from a book by Minnaert, "The Nature of Light in the Open Air," Dover, New York. It demonstrates the ability of the naked eye to percieve polarization in light and the orientation of the plane of polarization. To quote from Minnaert: "If you have a Nicol (prism) at your disposal,then look through it at a white cloud - or at an evenly illuminated (white) surface and try to distinguish the figure by the fact that the it revolves when the Nicol is rotated......." "Haidenger's brush is caused by the dichroism of the yellow spot on our retina. That all observers do not, apparently, see this remarkable figure in the same way no doubt depends on the difference in shape and structure of this yellow spot...." Hope this helps. All the best, Andy Andy Buechele The Catholic University of America 409 Hannan Hall Washington, D.C. 20064 (202) 319-4995 FAX: (202) 319-4469
We have recently received samples of freeze-dried guar seeds for SEM processing. The researcher would like the seeds cut in half and evaluated with SEM. In their current state, the seeds crumble when cut. If anyone has any suggestions for processing, we could use the help!
Thanks in advance, Beckie Anderson Kansas State University College of Veterinary Medicine Department of Diagnostic Medicine/Pathobiology Electron Microscopy Laboratory
Thank you to Massimo Sassaroli, Rober Mixon, Richard Mount, Stephan Coetzee, Stephen Griffiths, and others who responded to the query on how to segregate the MSA e-mail from the rest.
With the help, I was able to set the IE e-mail browser ("Internet Mail and News") to recognize the "microscopy" keyword in either the "TO:" or the "CC" - fields which are invisible by default. It has worked without a glitch for a week now, so no need to try a different browser. It mystifies me why it has worked in few circumstances as I'm unable to manually find the keyword in either of the fields - maybe "it's a feature and not a bug", but that's beside the point.
Electron Microscopy Sciences has Anhydrous Glut. (10% in acetone) in 10ml ampules, cat# 16530.
Tom
Thomas Moninger moninger-at-emiris.iaf.uiowa.edu University of Iowa Central Microscopy Research Facility http://www.uiowa.edu/~cemrf Views expressed are mine. On Fri, 7 Nov 1997, Rosemary White wrote:
} } Dear all, } } Does anyone know of a source of glutaraldehyde powder? We would like to } try glut in acetone and other solvents for freeze-substitution, but glut } comes only in ethanol as far as I can tell. I suppose we could dry it down } from 70% solution, or does this alter it chemically?? } } TIA, } } } Rosemary White } Department of Biological Sciences } Monash University, Melbourne, Victoria 3168, Australia } phone 61-3-9905 5670 } fax 61-3-9905 5613 email r.g.white-at-sci.monash.edu.au } }
Please post responses to the list; I want to be safe too! :) Karen
o.oshea-at-queens-belfast.ac.uk wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Would anyone know details of suppliers where Uranyl acetate can be obtained } already in solution i.e. a saturated solution in 50% ethanol. We previously } made this up ourselves from dry powder and ethanol, but we are trying to } reduce the potential hazzard to health by purchasing ready made solution. } Thanks on advance } Orla O'Shea, Dept of Anatomy, QUB } o.oshea-at-qub.ac.uk
-- Karen Zaruba kszaruba-at-mmm.com 3M Company, St. Paul, MN 55144 "Opinions above are my own, not necessarily my employer's"
I have a question maybe someone could help me... I want to measure the size of my probe, but I collected the wrong image... i.e., I collected the image of the probe in the diffraction mode... Is it possible to find actual size of the probe? I think if I know the distance between the objective aperture and the selected area aperture and the point of convergence of the convergence angle, I should be able to calculate it from there... but where can I find all these information?... Can anyone help me?
I've done preps of individual cells on filters before with no particular problems. This one has me stumped.
I'm trying to get a prep of Scenedesmus cells onto a filter membrane for fixation, CPD, sputtering. Seems straightforward. These cells fly away from the Nucleopore (PC) or Millipore (cellulose ester) membranes when I transfer to a liquid after depositing the cells by gentle suction. One protocol I tried called for poly-L-lysine treated glass, but the cells failed to stick to this also. The microbiologist who is involved tells me that the cell surface is nearly pure cellulose and does not have the substances that give a negative charge and would make the polylysine work. SOME cells do stick to the filter surface but many do not and we don't know if this gives a selection for a subset (and we want to see them all).
Are algae this different? Are there tricks with the polylysine? Is there another method?
If anyone has any suggestions that could help me I would really appreciate hearing them.
Thanks in advance!
Dale Callaham (dac-at-bio.umass.edu) +++++++++++++++++++++++++++ Dale A. Callaham Central Microscopy Facility The University of Massachusetts Amherst, MA 01003 +++++++++++++++++++++++++++
Minnesota Microscopy Society Meeting November 20, 1997, Thursday
Atomic Force Microscopy & Related Techniques: Introduction, Instrumentation & Application to Polymeric Materials by Inga Holl Musselman, Assistant Professor of Chemistry, University of Texas, Dallas
University of Minnesota, St. Paul Campus Student Center, The Pendergast Room
5:30 - 6:00 Social with Appetizers Cheese - Crackers - Hot Apple Cider
Atomic force microscopy (AFM) was introduced by Binnig, Quate and Gerber in 1986. In this method, a sample is scanned beneath a small probe attached to the apex of a flexible cantilever. Cantilever deflection is measured to give height information corresponding to the sample topography. Since AFM relies on tip-sample force interaction, the technique can be applied to insulators as well as to conducting and semiconducting materials. AFM therefore extends local probe studies to an important class of materials which can be difficult to investigate by electron microscopy and spectroscopy techniques owing to problems with sample charging.
During the past decade, related force microscopy methods have been developed to facilitate the study of surfaces in a variety of environments using a number of contrast mechanisms. Among others, these methods include contact, non-contact and TappingMode atomic force microscopy, lateral force microscopy, force modulation, phase imaging, electrostatic force microscopy, magnetic force microscopy, scanning capacitance microscopy, scanning near-field optical microscopy, and scanning near-field thermal microscopy. This presentation will review the theory and instrumentation for some of these microscopy methods and will emphasize their application to polymer materials.
Biography: Inga Holl Musselman is an Assistant Professor of Chemistry at the University of Texas at Dallas. She received a Ph.D. in Analytical Chemistry in 1988 from the University of North Carolina at Chapel Hill. Her Ph.D. research project, concerning molecular and quantitative aspects of laser microprobe mass spectrometry (LAMMS), was conducted at the National Institute of Standards and Technology. During a postdoc in the Department of Materials Science and Engineering and Precision Engineering Center at North Carolina State University, her research efforts concerned the fabrication of controlled geometry tips for scanning tunneling microscopy (STM) (patent and license awarded). In collaboration with Hoechst Celanese, she also investigated the application of scanned probe techniques to the characterization of polymer surfaces. Currently, Dr. Musselman's research group is investigating the fundamentals of STM image contrast. In addition, they are using atomic force microscopy and other microscopy methods to characterize the microstructure of synthetic and biopolymers including gas separation membranes and paired helical filaments from Alzheimers diseased brains.
PLEASE MAKE RESERVATIONS by November 18th. Contact: Gib Ahlstrand at (612)625-8249, 625-9728FAX, or: giba-at-puccini.crl.umn.edu.
Dinner and Social Hour: $10.00 per person, payable at the door. Free for current student members or with new sudent membership,payable at the door. Presentation is free for those who come later for the talk only.
Social hour, dinner and the presentation will all be held in the Pendergast Room, second floor level of the St. Paul Campus Dining Center (across hall from Cherrywood Room) of the University of Minnesota. This location will be familiar to some who have attended our meetings there before. Parking is available in the lots indicated with cross-hatching below:
Directions: From I-94, take I-280 a few miles north and exit onto Larpenteur Ave., go eastbound 1 mile to Cleveland Ave. plus 1 block to Gortner Ave. From I-35W or Hiway 36 take Cleveland Ave. south to Larpenteur, turn left go 1 block to Gortner Ave. Turn right onto Gortner and go south to Buford Ave., turn right , go 1 block to Buford Circle, turn right and proceed 1 block to parking lots (see map). Enter Dining Center as indicated.
--
Gib Ahlstrand, MMS Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } There is a description of the phenomenon of "Haidinger's brush" (or } cross) in "Condepts of Classical Optics" by John Strong, Freeman, San } Francisco, 1958, P.111f. He quotes from a book by Minnaert, "The } Nature of Light in the Open Air," Dover, New York. It demonstrates } the ability of the naked eye to percieve polarization in light and } the orientation of the plane of polarization. To quote from Minnaert: } "If you have a Nicol (prism) at your disposal,then look through it at } a white cloud - or at an evenly illuminated (white) surface and try } to distinguish the figure by the fact that the it revolves when the } Nicol is rotated......." } "Haidenger's brush is caused by the dichroism of the yellow spot } on our retina. That all observers do not, apparently, see this } remarkable figure in the same way no doubt depends on the difference } in shape and structure of this yellow spot...." } Hope this helps. } All the best, } Andy } Andy Buechele } The Catholic University of America } 409 Hannan Hall } Washington, D.C. 20064 } (202) 319-4995 FAX: (202) 319-4469 } There is another description of what is called an "Optic Axis Interference Figure" in Nesse, William D, Introduction to Optical Mineralogy, Second Edition 1991, Oxford university press. While I realize the application may be different, the figure is the same (caused the same way) and the book gives a somewhat detailed account of how the figure (cross) is formed, and what the colors (blue and yellow) and the positions of the colors in the cross mean. Along with a description of how these crosses are used to help identify minerals in petrographic sections (though most of you could probably care less :)).
Reichert makes an automatic grid stainer that uses prepared stains (called the "Ultrostainer"). Maybe they would be a potential supplier for premixed uranium stains.
Bob Wise } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Rosemary White wrote: } } } Dear all, } } Does anyone know of a source of glutaraldehyde powder? We would like to } try glut in acetone and other solvents for freeze-substitution, but glut } comes only in ethanol as far as I can tell. I suppose we could dry it down } from 70% solution, or does this alter it chemically?? }
Dear Rosemary,
Glutaraldehyde monomer is notoriously unstable in aqueoussolutions above 70%. Glut polymerizes at high concentraions and is next to impossible to depolymerize. For these reasons I believe it would be inadvisable to dehydrate your 70% Glut. We at Ladd currently offer 20% Glut in anhydrous methanol (catalog # 20257 & 20258). We have produced a wide variety of glutaraldehyde combinations through the years and would be willing to prepare some glut in acetone and in other solvents. If you are interested please contact me direct and we can work out the particulars.
Dr. Charles Duvic Ladd Research 13 Dorset Lane Williston, VT 05495 USA tel 1-802-878-6711 fax 1-802-878-8074 e-mail ladres-at-worldnet.att.net
I've recently been handed a sample consisting of dust collected near a suspected pollution source. The sample was picked up on cotton swabs in an extremely non-scientific manner (off the hood of a car in the desert, and they want to know if silicon is present---go figure), and the client is looking for fiberglass particles, among other things. I have found fibers in the sample about 10-12 microns in diameter which resemble SEM photos I have seen of fiberglass.
My question is this: What elements would one expect to find in fiberglass, other than (obviously) silicon? I'm aware of calcium and aluminum being used in glass, but are there other commonly used elements? Specifically, barium? (5 distinct Ba peaks in this one!)
The sample is on a carbon stub using carbon adhesive tape. We're using a variable pressure SEM with 7 Pa of pressure. The EDS on the fibers is being done in spot analysis mode at a mag of 3000x. I'm reasonably confident that most of the signal is coming from the fiber itself, allowing of course for beam skirting, etc.
Thanks in advance for any advice.
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
Thank you to all the people who responded to my question about how film thickness monitors work. I am now a lot more knowlegeable about the subject. All your replies were greatly appreciated.
The New England Society for Microscopy (NESM) presents its 31st Annual Fa= ll Symposium and Business Meeting on December 5, 1997 at the Hyatt Regency Cambridge, 575 Memorial Drive, Cambridge, MA 02139, Tel. (617) 492-1234.
PROGRAM: Friday, December 5, 1997
Session I =
9:00 am Registration and Continental Breakfast Note: The Registration Desk will close at 10:45 am and reopen for =
Session II from 1:00 to 1:30 pm
9:55 am Welcome =
Ann Hein, NESM President
10:00am "Characterization of Graphitic Carbon Spheres and Tubes Synthesized =
by a Mixed-Valent Oxide Catalytic Carbonization Process" Dr. Z. L. Wang School of Material Science and Engineering Georgia Institute of Technology
11:00am "Pathology of Laser Tissue Interaction" Thomas J. Flotte, MD Associate Professor of Dermatology, =
Harvard Medical School, Boston, MA Director of Dermatopathology, Massachusetts General Hospital =
12:15pm Luncheon with wine: Poached Atlantic Salmon, Chicken Picatta, or =
Vegetarian selections =
Session II =
1:00 pm Registration =
1:30 pm "Characterization of Translucent Polycrystalline Alumina Using SEM, =
Electron Probe and TEM" =
Dr. George Wei, Osram Sylvania
2:15 pm "Confocal Microscopy of Live Subjects" Robert H. Webb Wellman Laboratories of Photomedicine, MGH Schepens Eye Research Institute, Boston, MA
3:00 pm Coffee Break
3:15 pm "What Project MICRO Can Do For You" Caroline Schooley, MSA Education Outreach Coordinator =
NESM Annual Business Meeting 4:15 pm Counting of Ballots =
Reading of Minutes of 1996 Annual Business Meeting =
Report of the President Report of the Treasurer Introduction of New Officers =
5:00 pm Adjournment Louis Kerr, Incoming NESM President
For more information or to register, contact L. Kirstein at NESM-at-compuserve.com =
or by phone at (508) 473-9673. (Please include your fax number if you ha= ve not =
} The sample was picked up on cotton swabs in an } extremely non-scientific manner (off the hood of a car in the desert, and } they want to know if silicon is present---go figure)
Carnack says, "Yes!"
} ... the client is looking for fiberglass particles, among other things.
Randy, how can I put this? Um, "you're driving a thumb tack in with a sledge hammer?" Or, perhaps.... Nevermind. An SEM with EDXA is overkill for finding fiberglass. It is a trivial task with a polarized light microscope as all glass is transparent and isotropic and various other fibrous materials are either opaque or birefringent. In addition, the morphology will tell you if it is mineral wool, a course fibrous glass used most often in insulation, or true fiberglass, a more carefully prepared fibrous glass product used in a wider range of applications. The size you mention (10-12 microns) is certainly within the range one can encounter with either of these products, or other fibrous materials.
If you still have a bit of the swab(s) left, I would suggest the following. Place a portion of the soiled swab in a small centrifuge tube, add a bit of water with dilute detergent, ultasonicate for a few minutes, remove the swab, centrifuge, wash a couple of times, re-suspend and pipet a bit of the particulate debris onto a microscope slide. Mount in virtually anything and examine under a PLM with slightly uncrossed polars. Look for isotropic, fibrous strands. It takes longer to describe than it does to do!
} My question is this: What elements would one expect to find in fiberglass, } other than (obviously) silicon? I'm aware of calcium and aluminum being } used in glass, but are there other commonly used elements? Specifically, } barium? (5 distinct Ba peaks in this one!) }
But on the other hand, if an SEM is all you have....
As I mentioned, mineral wool is a fairly crude product. Consequently, it can contain a lot of "junk" elements, among them Ca, K, Na, Al, Mg, Fe, S, Cl. I don't know about barium but nothing would surprise me. Because of the elemental "ambiguity" of fibrous glass, PLM is really the way to go here.
Reference (of course): The Particle Atlas Electronic Edition.
--
Stephen A. Shaffer MicroDataware sshaffer-at-microdataware.com (business) http://www.microdataware.com (temporarily out of service) steve_shaffer-at-compuserve.com (personal) http://ourworld.compuserve.com/homepages/steve_shaffer/
We are looking for sources for rebuilt Filaments for a JEOL 5800LV SEM. Can anyone give us info on this?? (Where to get them and how much they are if possible).
Has anyone had experience with eliminating excessive amounts of dust in their EM rooms?
I moved into a newly renovated suite of rooms over a year ago. Since that time we have been plagued by dust and dirt. It has become almost impossible to maintain uncontaminated grids or negatives and prints without dust or scratches. For example you cannot leave a negative on a light box overnight because it is covered by a thin film of dust by the next day.
The Engineering department had the ductwork cleaned and higher efficiency filters installed on the fan units supplying air to the suite during renovations. They claim the supply air is the same quality as that in my previous location in another building (where excessive dust was not a problem).
An outside consultant has monitored the rooms and found particle counts between 20,000 and 50,000 in the 0.5 micron size range, per cubic foot. They recommend installing HEPA filters on all supply air ducts to reach class 1,000 conditions.
The Engineering department is reluctant to install 10 or more HEPA filter units due to the cost and the necessary ductwork reworking. They claim the supply air is clean and the filters will not solve the problem. They would like to install electrostatic particle precipitators in each room to scrub the air clean.
Does anybody have any experience with electrostatic precipitators and which type is best?
Is there a standard for airborne particles within a microscopy facility?
Is the class 1,000 condition recommended by the consultant excessive?
We are about to begin renovations for our light microscope area which will have a number of digital imaging stations in addition to film and video. What recommendations can I make to engineering in the design of the HVAC system to address this issue during construction?
Sorry for the excessive bandwidth of this message but I wanted to be as explicit as possible.
Thanks in advance for your help, Frank Macaluso **************************************************************************** Frank Macaluso tel: 718-430-3547 Analytical Imaging Facility fax: 718-430-8996 Albert Einstein College of Medicine e-mail: macaluso-at-aecom.yu.edu 1300 Morris Park Avenue Bronx, NY 10461 ****************************************************************************
} Has anyone had experience with eliminating excessive amounts of dust in } their EM rooms? }
Frank, better responses may be forthcoming from others, but one quick thought occured to me. You might ask your Engineers if the return air passes through a "plenum" above the ceiling or below the floor. This is just a fancy word for saying that the open space (above or below) acts as the return duct. If this is the case, and it often is, that might be the source of a large part of your problem. You've probably seen, or can imagine, the cleanliness of these areas! If so, regardless of filtration systems, you should probably start with a dedicated HVAC system for the microscopy rooms and incorporating duct work for both supply and return sides. Then add filtration as necessary to achieve acceptable cleanliness. It may be that, in your circumstances, Class 1000 is more than really necessary but you're probably in for some expensive retrofitting in any event.
--
Stephen A. Shaffer MicroDataware sshaffer-at-microdataware.com (business) http://www.microdataware.com (temporarily out of service) steve_shaffer-at-compuserve.com (personal) http://ourworld.compuserve.com/homepages/steve_shaffer/
Dale Callaham wrote: =================================================== I'm trying to get a prep of Scenedesmus cells onto a filter membrane for fixation, CPD, sputtering. Seems straightforward. These cells fly away from the Nucleopore (PC) or Millipore (cellulose ester) membranes when I transfer to a liquid after depositing the cells by gentle suction =================================================== While there are no guarantees, you might want to consider the following:
a) Polycarbonate "track etch" (PCTE) membrane filters, irrespective of who has made them, including the SPI-Pore(tm) membrane filters are treated with PVP (polyvinylpyrrolidone) to serve as a wetting agent, making the surface more hydrophilic and enhancing the filtration rates. We have special requests for supplying the membrane filters without PVP (for several different reasons) which can be done and apparently, in some instances, affects the way things adhere. Not wanting to be too commercial about it, it is my understanding that at least one other supplier of PCTE membrane filters can supply their products without PVP as well. In order not to mislead, there are in fact different production processes in use and not all PCTE membrane filters are identical. This is an instance where if you have tried one, you have certainly not tried them all. I am not saying this is a matter of good vs. bad, this is just a matter of membrane filters from different sources can be expected to behave "different".
b) Silver membrane filters also seem, at least in some instances, to provide better attachment possiblities than the polymer membranes. I have been intrigued as to why that is the case, but the silver membranes can be handled in a surprisingly similar way as are the polymer membrane filters, anything you can do, including critical point drying, to a polymer membrane you can do to a silver membrane. A bonus with the use of silver membrane filters is that much less metallization or no metallization is required. I have also been told that there are indications that a plasma etching ("cleaning"), presumably because of a removal of adsorbed organics, also enhanced cell adhesion.
Disclaimer: SPI offers PC and silver membrane filters as well as the a plasma etcher, details about which can be found on our website below.
Chuck =================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Currently, I am searching for protocols that deal with collecting stool samples and preparing them for TEM. Does anyone have such an item? My purpose for using this protocol is to eventually isolate viral structure in patients with Acute Disseminated Encephalomyelitis.
Thank you, Debra Caires The National Encephalitis Foundation
San Jose State University Biological Sciences One Washington Square San Jose, CA 95192 =46AX (408) 298-3263
The proceedings of EUREM-11 (Dublin 1996) will shortly be published by CESM, with the full backing of IFSEM. The 3-volume set will be available for a maximum price of 400 French Francs (approx US$ 70); the exact price will depend on the number of orders received and since the books will be printed in France, the price in francs is definitive.
If you are interested, please request an order form from Peter Hawkes E-mail: hawkes -at-cict.fr or hawkes-at-cemes.fr Fax: (+33) 562 25 79 99
Be sure to give your fax number or full postal address. Please respond IMMEDIATELY as we hope to print these proceedings very soon indeed and only a limited number will be available once orders have been serviced.
Dr Peter W. Hawkes CEMES-LOE du CNRS B.P. 4347 31055 TOULOUSE cedex 4 (France)
Dear Randy, In my experience in looking at fiberglass, I would expect some Na, which is a common part of most glasses. The Ba may be from BaSO4, which is a common whitener or may also be in glass. The fiberglass fibers should be unnaturally smooth and uniform. Rock wool, which was also mentioned, is very coarse, more like 20 microns, and dark in color. You wrote: } Greetings to All, } } I've recently been handed a sample consisting of dust collected near a } suspected pollution source. The sample was picked up on cotton swabs in an } extremely non-scientific manner (off the hood of a car in the desert, and } they want to know if silicon is present---go figure), and the client is } looking for fiberglass particles, among other things. I have found fibers } in the sample about 10-12 microns in diameter which resemble SEM photos I } have seen of fiberglass. } } My question is this: What elements would one expect to find in fiberglass, } other than (obviously) silicon? I'm aware of calcium and aluminum being } used in glass, but are there other commonly used elements? Specifically, } barium? (5 distinct Ba peaks in this one!) } } The sample is on a carbon stub using carbon adhesive tape. We're using a } variable pressure SEM with 7 Pa of pressure. The EDS on the fibers is } being done in spot analysis mode at a mag of 3000x. I'm reasonably } confident that most of the signal is coming from the fiber itself, allowing } of course for beam skirting, etc. } } Thanks in advance for any advice. } } } Randy Tindall
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 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
Thankyou to those who replied to my query re pre-prepared Uranyl Acetate, I was asked to post replies to server and here they are. Charles Garber Presidentof SPI Supplies wrote that his company had considered pre-prepared uranyl ace- tate some years ago but found the cost to be extremely high, another suggestion was that as Reichert make an automatic grid stainer, they may be able to supply uranyl acetate in this form. Thanks again, Orla O'Shea.
I am interested in looking at tape heads used in digital data storage systems via a light interferometry system. We wish to measure parameters such as head profiles, gap widths and other issues to less than a micron resolution. To do this accurately we need to know the refractive index of the different materials in the heads. Some of them are exotic alloys which makes this difficult, and I am trying to think of alternative techniques to assist us. Coating the samples would be one idea since the light would be effectively incident upon the same material but they have a rather fiddly cross section and I am not convinced that we could coat the samples without any shadowing. Replication seemed like a good idea, apart from the resolution worries, or would we have the same shadowing problems? Does anyone have any knowledge about this?
It is not uncommon to findphosphates and borates in significantly high quantities in the fibreglass materials. As to, morphological identification anisotropy of fibres makes it easy to identify. However beware that fibres morphology may contain gas bubbles. So btoken fibres may look jagged with curved edges
At 01:44 PM 11/10/97 -0700, Christopher wrote: } We are looking for sources for rebuilt Filaments for a JEOL 5800LV SEM. } Can anyone give us info on this?? (Where to get them and how much they } are if possible).
Energy Beam Sciences has been manufacturing both new and rebuilt filaments for JEOL SEMs for more than 25 years. I will respond to the pricing question directly to Christopher and hope the other manufacturers will follow the same policy {grin}
Best regards, steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
I do it rountinely in my EM lab for animal clinics.
-use a disposal pipe to pick a tiny sample and dispense in a vial containing 0.5-1 ml of 0.1% glutaraldehyde in buffer, mix the sample well and let it sits for 15-30 min.
-do negative staining as usual from the supernatant in 1-2% PTA.
Good luck.
On Mon, 10 Nov 1997, Debra J. Caires wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear All, } } Currently, I am searching for protocols that deal with collecting } stool samples and preparing them for TEM. Does anyone have such an item? } My purpose for using this protocol is to eventually isolate viral structure } in patients with Acute Disseminated Encephalomyelitis. } } Thank you, } Debra Caires } The National Encephalitis Foundation } } San Jose State University } Biological Sciences } One Washington Square } San Jose, CA 95192 } FAX (408) 298-3263 } } } } }
*********************************************** * Ming H. Chen, PhD * * Medicine/Dentistry Electron Microscopy Unit * * University Of Alberta. * * Edmonton, Alberta, Canada * * * * Visit My Page At: * * http://www.ualberta.ca/~mingchen * ***********************************************
I am finding that lifetime for my FEI LaB6 cathode exceed 1000hrs, but after 100hr the gun becomes unstable (... constant tilt/shift alignments ...) to the point I have to get into the gun and clean the wehnelt, which remedies the problem. The deposits are difficult to remove and I have to resort to judicious use of diamond paste. I'd much rather find an agent which would remove these deposits over-night and not attack the SS metal. Has anyone a better idea??? ... TIA ...
cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
Dear Sirs I am an undergraduate Physicist at King's College London. As a final year student, we are expected to complete a major laboratory project.We have come up against a problem which we feel is beyond our capabilities. In order to measure accurately the thickness of some thick films we have made, we need to interface our TEM with the P.C, via a printer board electronic circiut. We as physics undergrads are unable to write the correct syntax for the programme for the interface and would be grateful if you could help us with it. If your archive has the bit we need in "C" please e-mail it to me. If you are unable to help, thankyou anyway. Regards, Jennifer Ayriss.
} -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Your are right, the background is not going to be smooth structureless and } featureless like on a TE membrane. However, I have seen fragile features on } certain life science samples that could not survive the level of } metallization that would other wise be required. I was addressing my } comments to those kinds of situatons. } } What do you think about the belief that there is better attachment to non- } PVP treated membranes, is that possible or just an old wive's tale? } } Chuck
Hi Chuck, I can't comment on the above belief. You are the first who has mentioned it to me. I'd have to conduct an empirical comparison between treated and non-treated, I'd also include poly-L-lysine and collagen treated membranes as long as I was going to the trouble. Has any of the collective ever looked at this?
I haven't put any metal on an SEM sample in about a year, I've been doing uncoated LVFESEM with pretty decent results. Of course I'm not going up to 50,000x either. I need to be able to shuttle back and forth between SEM and TOF-SIMS instruments.
I do pre-coat some PC membranes with AuPd for conductivity prior to applying samples. Again, I don't know what adsorbtion changes may be induced. Particulate samples (yeast) are still present after freeze drying.
cheers ed
Edward J. Basgall, PhD The Pennsylvania State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618 http://www.personal.psu.edu/ejb11/ Privilege does not absolve one of ecological responsibility.
} Dear All, } } Currently, I am searching for protocols that deal with collecting } stool samples and preparing them for TEM. Does anyone have such an item? } My purpose for using this protocol is to eventually isolate viral structure } in patients with Acute Disseminated Encephalomyelitis. } } Thank you, } Debra Caires } The National Encephalitis Foundation } } San Jose State University } Biological Sciences } One Washington Square } San Jose, CA 95192 } FAX (408) 298-3263 } Hi Deb, There are numerous references available on this. I did a poster on the subject at MSA in 1988, Basgall, E.J., Scherba,G., and Gelberg, H.B. "Diagnostic Virology in Veterinary Pathology: Techniques for Negative Staining." I will FAX you a copy of the abstract with references today.
good luck cheers ed
Edward J. Basgall, PhD The Pennsylvania State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618 http://www.personal.psu.edu/ejb11/ Privilege does not absolve one of ecological responsibility.
At 08:14 AM 11/11/97 -0800, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 352-392-1295 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
} } } Ed - } } Did you make the "bread" part of the sandwich yourself? A lab I } } worked in ages ago had special Millipore snap-together units to do this } } same thing; I've never been able to track them down from any vendors. i } } would like to see how you do this - the homemade sandwiches I've tried } } always loosen partway through fixation. } } } } Thanks! } } } } Tamara Howard } } CSHL } } Yes Tamara, } } I remember those snap together units. Thomas Scientific (800-345-2100) } still sells them, CAT # 4626-N20 13mm, p585 in 96-97 catalog. 10/pk list } price $30. Made from polycarbonate plastic. } } No affiliation or financial interest, yadda, yadda, yadda.... } } I will have the filter sandwich details posted to my website by Friday (11-14) } } cheers } ed } } Edward J. Basgall, PhD } The Pennsylvania State University } Surface Chemistry Group ejb11-at-psu.edu } Materials Research Institute Building Ph: 814-865-0493 } University Park, PA 16802-7003 FAX: 814-863-0618 } http://www.personal.psu.edu/ejb11/ }
} i have archived a recent discussion on this at the following URL (Tips & } Tricks site): } } http://www.biotech.ufl.edu/icbr/emcl/db/lab6crud.html } } ...
Thanx ... it appears the consensus is dilute HCl for short periods together with H2O rinses, and I can't imagine this being much of a problem for stainless steel ... but it *was* strange to see someone else suggest ammonia ... that is, base over an acid attack ... and can I assume the "soap-scum remover" suggestion is also basic. I can't imagine ammonia being a problem either, 'cept its suggested use is an over-nite bath ...
thanx again, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
} i have archived a recent discussion on this at the following URL (Tips & } Tricks site): } } http://www.biotech.ufl.edu/icbr/emcl/db/lab6crud.html } } ...
Thanx ... it appears the consensus is dilute HCl for short periods together with H2O rinses, and I can't imagine this being much of a problem for stainless steel ... but it *was* strange to see someone else suggest ammonia ... that is, base over an acid attack ... and can I assume the "soap-scum remover" suggestion is also basic. I can't imagine ammonia being a problem either, 'cept its suggested use is an over-nite bath ...
thanx again, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
The site listed below is an expert system for determining cleaning procedures. Great stuff. If dilute HCL doesn't work, then this web site might a good place to get direction.
http://clean.rti.org/
------------------------------------------------ Opinions or statements expressed herein, rational or otherwise, do not necessarily reflect those of my employer.
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 Phone: (508) 750-1717 E-mail: crossman-at-osi.sylvania.com
Our web sites: www.sylvania.com www.siemens.com --
Another alternative you may consider still used the polish paste. Our sevice eng. taught us this one. Take a dremel and put a Q-tip in it (drill may work too) put on some paste and let her rip. We don't run lab6 so I have no experience with that aspect, just the usual crud in any biologic EM.
At 10:05 AM 11/11/97 -0800, you wrote: } Scott Whittaker wrote: } } } i have archived a recent discussion on this at the following URL (Tips & } } Tricks site): } } } } http://www.biotech.ufl.edu/icbr/emcl/db/lab6crud.html } } } } ... } } Thanx ... it appears the consensus is dilute HCl for short periods together } with H2O rinses, and I can't imagine this being much of a problem for } stainless steel ... but it *was* strange to see someone else suggest ammonia } ... that is, base over an acid attack ... and can I assume the "soap-scum } remover" suggestion is also basic. I can't imagine ammonia being a problem } either, 'cept its suggested use is an over-nite bath ... } } thanx again, shAf } -- } {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} } Michael Shaffer, R.A. - University of Oregon Electron Probe Facility } mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu } http://darkwing.uoregon.edu/~mshaf/ } } } }
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall Research Assistant Gainesville, FL 32610 University Of Florida ph 352-392-1295 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
} Another alternative you may consider still used the polish paste. Our } service eng. taught us this one. } Take a dremel and put a Q-tip in it (drill may work too) put on some paste and } let her rip. ...
My service rep reccommended against this as it can be quite abrasive. However, the technique is okay if you buy the variable speed dremel and use its slow speed ... saves quite a bit of elbow grease ... cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
} Another alternative you may consider still used the polish paste. Our } sevice eng. taught us this one. } Take a dremel and put a Q-tip in it (drill may work too) put on some paste } and let her rip. We don't run lab6 so I have no experience with that } aspect, just the usual crud in any biologic EM.
Be careful if you use this technique because it can quickly enlarge and distort the aperture of the Wehnelt shield or cap. Once enlarged, the gun geometry is degraded until you replace the cap with a new one.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
We have been using the ultrasonic bath with "Micro" cleaning solution (Catalog #6731, International Products Corp, 609-386-8770) to clean the wehnelt, apertures, etc.
I used to use the metal polish and much scrubbing with q-tips, cotton, followed by ultrsonic rinsing with isopropanol. This cleaning solution does a great job, and removes tungsten deposits that I could not previously remove.
The solution works best when warm, and I must confess that I use a stronger mix than the instructions dictate (instructions say 2%, I use about 10% or so in distilled water). I put this brew into a big glass beaker, heat it up on the hot plate, put in the parts, and immerse in the ultrasonic bath. I follow that by sonication in distilled water with a finish using isopropanol. We just did a column routine on our Jeol 733 using the solution and it also removed deposits from little crevices that were not cleaned that well before.
I have not observed any detrimental effects on the pieces we have cleaned, and am especially interested in hearing about it if it does happen.
The solution smells of ammonia but the ingredients show more than that.
I think I paid about $10 for 1 quart. Highly recommended.
Paul Carpenter
+----------------------------------------------------+ | Paul K. Carpenter paulc-at-gps.caltech.edu | | Division Analytical Facility | | Geological and Planetary Sciences MC 100-23 | | California Institute of Technology | | 1200 East California Blvd | | Pasadena, CA 91125 | | 626-395-6126 (X-ray Lab) 626-568-0935 (Dept. FAX) | +----------------------------------------------------+
Ciara: If you can rotate the specimen during C evaporation rather complex specimens can be coated with no 'shadows' remaining. The replication technique too should work. Plastic replicas can cope with difficult shapes too and they are quite easy to produce. A basic outline of the process is given under 'replication' (see index or "V" page) in our online catalogue. Replicating materials and instructions are provided by us and I expect all other EM suppliers. You would probably later angle shadow the replica with metal to show depths. With a known angle you can estimate height and the incorporation of latex spheres would provide an internal scale. Plastic replicas are accurate to better than 0.1um. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au
---------- } Hello } } I am interested in looking at tape heads used in digital data storage systems } via a light interferometry system. We wish to measure parameters such as head } profiles, gap widths and other issues to less than a micron resolution. To do } this accurately we need to know the refractive index of the different materials } in the heads. Some of them are exotic alloys which makes this difficult, and I } am trying to think of alternative techniques to assist us. Coating the samples } would be one idea since the light would be effectively incident upon the same } material but they have a rather fiddly cross section and I am not convinced that } we could coat the samples without any shadowing. Replication seemed like a } good idea, apart from the resolution worries, or would we have the same } shadowing problems? Does anyone have any knowledge about this? } } } Thanks in advance } } Ciara } } } } *********** *********** Dr C A Mullan } ******** / ******** Computer Peripherals Bristol, } ****** /__ __ ****** Hewlett-Packard Ltd, } ***** / / / / ***** Filton Road, Stoke Gifford, } ***** / / /__/ ***** Bristol, England. BS12 6QZ } ****** / ****** } ******** / ******** } *********** *********** } Tel: +44 (0) 117 922 9908 } Fax: +44 (0) 117 923 6091 } } } }
Be careful of electrostatic precipitators. They may produce low concentrations of ozone and contribute to rapid deterioration of rubber parts of equipment. Best test is the lifetime of a stretched rubber band; compare to control prep'n perhaps at home.
Air-conditioner type filters treated with a polyethylene glycol spray may stop the dirt to a great extent without going to the expense of HEPA filters. Under the conditions you describe, the initial cost might not be as big a burden as the cost of frequent replacements!
San Francisco Microscopical Society November Meeting Announcement
Annual Swap Meet November 15, 1997 =
=
Place: Rockridge Branch Oakland Public Library 5366 College Avenue Oakland CA
Time: 10:00 A.M.
Everyone has old microscope parts, equipment, supplies, and associated items that they would like to get rid of =96 and all microscopists have room in the microscopical cabinets for that perfect gizmo that the next persons wants to be rid of. This is your opportunity to get rid of those things you don=92t want any more, and replace them with those invaluable items that other people will bring.
Bring all of you tradeables to the meeting on Saturday and see what you can get rid off, and see what you can take home with you.
For further information, you may contact Peter Barnett at 510-222-8883 or visit the SFMS Meeting Web Page at: http://ourworld.compuserve.com/homepages/steve_shaffer/announce.htm The web page contains further information, maps, directions via car and public transportation, etc.
-- =
Stephen A. Shaffer MicroDataware sshaffer-at-microdataware.com (business) http://www.microdataware.com (temporarily out of service) steve_shaffer-at-compuserve.com (personal) http://ourworld.compuserve.com/homepages/steve_shaffer/
For those interested, the authors and titles of presentations to be given at the Microscopy Society of Southern Africa's 37th Annual Conference is available at:
http://www.uct.ac.za/depts/emu/mssa/contents.htm
Abstracts of these presentations are published in the conference proceedings.
The conference will take place from December 2 to 5, 1997, at the University of the Western Cape in Bellville, near Cape Town.
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa eurc-at-giraffe.ru.ac.za - tel: +27 461 318168 - fax: +27 461 24377
I am looking for a human pan-keratin antibody to recognise both epidermal and hair keratinocites. Can somebody suggest me a good, possible, policlonal antibody?
Dr. Cristiano Rumio Istitute of Human Anatomy Via Mangiagalli 31 20133 Milan Italy Tel. -39-2-2663683 Fax. -39-2-2364082 E-mail: crylsm-at-imiucca.csi.unimi.it URL: http://imiucca.csi.unimi.it/~endomi/confocal.html __________________________________________________________________________
I am busy with a literature study on STM/AFM and epitaxial grown Au layers. I have a few questions on using gold substrates in STM/AFM.
1. If you use gold evaporated onto mica as a substrate, do you need to separate the gold and mica before you can use the atomically flat gold as a substrate? Or do you stick it ont some kind of stub for the STM? 2. Is there a minimum/maximum thickness of gold foils suitable for substrates? 3. If you want to use atomically flat gold as substrate, do you buy it or make it yourself (and to all the vendors, I'm not interested in buying, just asking...).
Disclamer: I am NOT a vendor, I sell nothing (except perhaps my soul), I have done my best to come up with reasonable current market pricing in all the following comparisons and this is a LONG email.
Yes, high resolution images (which maintain their resolution, i.e. not subjected to 'losey-compresion') do take up alot of storage space - as my students are constantly shocked at. Storage of these images is problematic, and presently there is no ideal soloution, but here are some for consideration:
Definitions: Mb= Megabit (Divide by 8 to get MB), MB = MegaByte, GB=Gigabyte, 1000MB = 1GB (Hey some of us are really just starting out) and $ = U.S.D.
In order to make some sense in comparison I have choosen the simplistic view in regards to data transfer rates (i.e. the speed for transfering data between the storage device and the computer system/software) and simply have relied on manufacturers reported transfer rates. Things to keep in mind with these numbers (1) you'll never see these speeds in reality - they are all determined in ideal situations not real world usage, but they are comparable with each other; (2) Actual transfer rates will vary depending on the interface used (i.e. the max. throughputs for the interfaces are EIDE (1-10MB/sec) vs SCSI (5-10MB/sec) vs SCSI-Wide (20MB/Sec) vs SCSI-UltraWide (40MB/Sec) vs parallel ports ( {1 vs Fiber Coupling (a.k.a. Fire Wire, 100+MB/Sec), (3) System configuration will also effect this; i.e. what CPU, what BUS speed, how much memory (RAM) what other components are in the system, what operating system, etc. , (4) the biggest factor in the data transfer numbers game is "Maximum Burst" transfer speed vs "Throughput speed". My only hope in presenting this information is that is gives you a starting point for comparisons.
1) Zip drives are cheap ($140 for drive) but cartridges are expensive ($13 / 100MB - I'll use 100MB as the standard unit for price comparison). 100MB is not very much storage capacity, and the transfer speeds are very slow -at- 1MB/Sec NOTE: unless you have min. 650MB free HD space you can NOT record an entire CD directly (On -The-Fly) from a a zip drive, you'll have to record in multisession mode (and lose 13MB in "overhead" for each session after the first one, i.e. a 640MB CD-R would require 6 Sessions thus loosing 65 MB).
2) Iomega JAZ drives: $300/400 (Int/Ext) with 6.73MB/sec transfer, 1GB cartridge-at-$124 [$12/100MB] (Expensive AND slow!)
3) Don't be so quick to reject tape storage. Tape storage can be extremely cost effective in larger formats, speed can be a problem but going back to older work-around solutions, users wishing to work on files on Tuesday could transfer the data from tape to HD Monday night, and then transfer back - alternately, utilizing a file server for this task (File Server: HD's, Tape drives, CD-drives for handleing just data storage: reading and writing only, no running of other software) would free up workstations for computational work. Yes, random access is limited but if you're looking at serial section reconstruction you'll be accessing sequentially anyway. CON: You have to a have a drive to read and write the data (unlike ubiquitous CD-ROM readers).
Drives go upto the following: 20-40GB, 32-64GB, 70-150GB, 100-200GB, 140-280GB -at- $2,000-9,000.
NEW Iomega Ditto Max drives: upto 7GB* drives -at- $200, upto 10GB* drives -at- $300, (*these are compressed data values, compression will slow down transfer speeds) with transfer speeds upto 36MB/sec and tapes running $20/3gb - $35/10GB [$0.20-$0.30/100MB].
4) CD-ROM storage: This makes alot of sense, since 99% of all computers come with CD-ROM readers. If you (or your users) have older CD-ROM readers, then they will need to cough up $80-150 to buy a newer one (10x IDE drive - 16x SCSI). For comparison sake a 1x refers to the standard speed of an Audio CD and allows for 150kilbytes of data transfered per second. Therefore a 10x CD-ROM would allow for upto 1.5MB/second transfer. Actual transfer rates will vary depending on the interface used (i.e. IDE vs SCSI, etc.).
CD-Recorders (CD-R's): these vary ALOT in recording speeds (1x-4x -6x?) and prices follow ($340 - $800), plus you'll need recording software ($50-100), and its recommended that you have a dedicated temporary storage drive for the higher recording speeds ($180-500). CD-R's are sensitive to recording errors, but they have become much more routinely reliable in the last 1-2years (particularly with a dedicated temp storage disk). CD-R's are WORM disks (Write Once Read Many), that means if you record practice images, you can't erase them and re-use the space. However, the recording media is reasonably cheap -at- 3.50-5.00/640MB disk [$0.54-0.78/100MB].
CD-ReWriteables (CD-RW): These are new to market as of this spring they do allow for re-writing to the media. There are still only a few manufactures and they record at 1-2x presently and cost ~$500. The media is also very expensive ($25/disk) but is expected to drop too reasonable prices by early next year. However! CD-RW's apparently can only be read in CD-RW drives and NOT normal CD-ROM drives. Ricoh does offer a drive which fuctions as CD-ROM, CD-R, and CD-CW (MediaMaster ~$550).
5) DVD anyone? (and NO "DVD" is not a an anacronym, it once was but had three different definitions so the powers that be decided to just leave it as DVD). This ones easy: NOT READY FOR PRIME TIME YET. Currently readers only are available, and recorders MIGHT be come available late 1998, however the major manufactiring groups have had a falling out again after coming up with a "standard" recording format, and there are two incompatible (?) formats heading our way so it might be until 1999-2000 before this becomes a reality and affordable. However, it may offer the best solution to todays image storage problems, in that the DVD storage capacity is 4.7GB for single sided and 7GB for double sided. But we shall see, eh?
6) Optical Drives: Currently available optical dirves are ReWritable (there are still some WORM drives though) fall into two major categories: Mageneto-Optical (MO) drives and PD Drives. MO drives come in various sizes from 128MB upto 4.6GB, I would suggest that only drive 640MB and larger be considered. PD drives come in 640-650MB sizes, these sizes are very nice because they match the CD-R size. Therefore you could use the MO or PD for temporary storage and then archive to CD-R. However, once again you need to have a drive to read these MO or PD's as well as write them. Most of the drives will handle the next 2 or 3 sizes smaller capacity disks, i.e. the 2.6GB MO's will read/write 1.3 & 650 disks. Prices are as follows (Internal/External):
4.6GB, $1,450/1,550, disks -at- $99 [$2.15/100MB] [MediaStore has the Mitsubishi 4200 4.6GB on sale for $999! 4.2MB/sec transfer]
640/650 PD drives: $350/440, disk -at- $34 [$5.20/100MB]
Toray has a 650 PD, 900kilobyte/sec which is also a CD-ROM reader for $315/405.
7) Removable hard disk frames: Another alternative, one which has been choosen by Hollywood (See Advanced Imaging October 1997, P.74) is the usage of removable harddrives. Frames for making HD's removable run: $20:IDE; $30:SCSI; $70:SCSI-Wide. However this does not include the cost of the drive (obviously). Secondly, HD's are delicate instruments and care must be taken when moving them about, but a rack close to a user accessible computer system (say one with a CD-Recorder?) would be a useful solution for the highest speed data transfer. 2-6GB IDE's -at- $7-5/100MB, 2-10GB SCSI -at- $7-10/100MB, 2-23GB SCSI Ultra-Wides -at- $7-20/100MB (On all HD's Bigger drives have lowest costs per MB).
There are a few other, less common solutions, and each faclity will need to develope their own prefered solution. However, I'm sure there are a number of vendors out there who'd love to sell you a turn-key system package for $20-40k.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"640K ought to be enough for anybody." -- Bill Gates, 1981
Daraporn, I think the answer is no. I am assuming that you took a diffraction pattern with the probe focused on the sample (i.e. you have a BF CBED disk). In this case, you have only measured the convergence angle of the probe. If you had an infinitely small source, knew you were in focus and fully stigmated, and knew the Cs of the probe forming lens, you could determine the theoretic probe size. But, for a non-FEG TEM, the probe size is mainly determined by the demagnification of the finite source (i.e. the spot size selected). You might be able to go back and image probe under similar conditions to get an approximate value for the probe.
Hope this helps, -- Ray D. Twesten Center For Microanalysis of Materials (217) 244-6177 University of Illinois (217) 244-2278 (fax) 104 S. Goodwin Ave. (217) 359-4035 (home) Urbana, IL 61801
Daraporn Arayasantiparb wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear All, } } I have a question maybe someone could help me... I want to measure } the size of my probe, but I collected the wrong image... i.e., I collected } the image of the probe in the diffraction mode... Is it possible to find } actual size of the probe? I think if I know the distance between the } objective aperture and the selected area aperture and the point of } convergence of the convergence angle, I should be able to calculate it } from there... but where can I find all these information?... Can anyone } help me? } } Your suggestions is greatly appreciated. } } Sincerely, } Daraporn Arayasantiparb
You say that an outside consultant reports that your EM room averages 20K - 50K particles in the 0.5 micron range per cubic foot. From experience, I would be surprised if you get a visible film of dust observed on a light background overnight, but perhaps a visible film of dust on a dark shiny negative as you describe sounds reasonable, especially when viewed by scattered light. You definitely shouldn't get anything you can feel with your fingers overnight.
I recommend the article "Particulate Fallout Predictions for Clean Rooms" by Otto Hamburg in The Jour. of Environmental Sciences, May/June 1982 pp.15-20.
You can set out "witness plates" for various lengths of time and observe the amount of fallout you get and compare these results with environments of different load suspensions. Your witness plates should be identical to the substrates of interest....TEM grids and negatives, and be in the same place in your room. This can make a big difference. Particulate characteristics, air flow, humidity, substrates, etc. will be different from one person's environment to another's, meaning you cannot readily expect one person's experiences to be applicable to yours.
We have a particle prep lab that ranges between 1K-50K particles } = 0.5 micron per cubic foot, depending where you are and what phase the moon is in. We then have { Class 10 areas embedded within that where samples may be exposed to air. (Assuming that my laser counter's calibration is still good). Within the 20-50K areas that would be somewhat similar to yours, I would not expect to find significant dust accumulation on negatives overnight. However, I would guess that if the air was very dry, and there was plenty of air flow across the negatives, perhaps they would electrostatically collect enough by the next day to be a problem. I would not leave TEM grids out in this air if I was concerned about contamination.
Typical unfiltered office and home air where there is no smoke has about a few billion particles } = 0.5 microns per cubic foot. So if you have a rough idea of how rapidly dust accumulates there, you can apply the rules of 10 to get a VERY ROUGH idea of what you would get in the same environment if it had a few orders of magnitude less particulate. Again, I repeat, very rough.
I also have a story. I once worked in a cleanroom that had about 10,000 particles } = 0.5 micron per cubic foot in most areas (Class 10,000), and some Class 10 areas. About once every several days, I would find an enormous amount of dust on everything in the Class 10,000 areas and the clean room would have to be thoroughly cleaned. It appeared to be caused by "burps" in the plenum structure, caused by momentary pressure differentials, that allowed dust that had accumulated there to burp through seams in the ceiling. Solution: seal the seams. So your average dust suspension measurements may not reflect momentary, but significant, surges, for whatever reason.
Other useful references are:
FED-STD-209E Federal Standard for Clean Room and Work Station Requirements for Controlled Environments
MIL-STD 1246 Military Standard for Product Cleanliness Levels
Hope this helps.
Cynthia J. Zeissler Physical Scientist National Institute of Standards and Technology cynthia.zeissler-at-nist.gov 301-975-3910
Dear Frank, } } Has anyone had experience with eliminating excessive amounts of dust in } their EM rooms? } } I moved into a newly renovated suite of rooms over a year ago. Since that } time we have been plagued by dust and dirt. } Since you are now experiencing a problem, the simplest thing would be to put something like cheesecloth (Optic-wipe cloth works for us) over the AC input & return ports. This will cut down on the air flow, but will remove a lot of particles. If this works, there may be no need for more expensive remedies, and the worst case will be insufficient improvement. } } Does anybody have any experience with electrostatic precipitators and which } type is best? } My father had a dust allergy, and we had a precipitator in his room. They are very efficient at removing particles, but the air has to flow through the plates, so in your case they would have to be installed in the AC inputs. (It's different conditions when the particles are continuously introduced than when they are stirred up off the floor.) Furthermore, the ozone production already mentioned is a problem; activated charcoal filters after the precipitators could solve it. If you go this route, there will have to be regular maintenance of both the precipitators & the filters. Good luck. Yours, Bill Tivol
Richard E. Edelmann, Ph.D. wrote: } } 4) CD-ROM storage: This makes alot of sense, since 99% of all } computers come with CD-ROM readers. If you (or your users) have } older CD-ROM readers, then they will need to cough up $80-150 to buy } a newer one (10x IDE drive - 16x SCSI). } I don't get why users would have to buy a 10x IDE or 16x SCSI drive. Can't the CD's be written in a standard format that can be read at any speed?? I am about to buy a CD writer and apparently have missed something here. TIA, Tom
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
You say that an outside consultant reports that your EM room averages 20K - 50K particles in the 0.5 micron range per cubic foot. From experience, I would be surprised if you get a visible film of dust observed on a light background overnight, but perhaps a visible film of dust on a dark shiny negative as you describe sounds reasonable, especially when viewed by scattered light. You definitely shouldn't get anything you can feel with your fingers overnight.
I recommend the article "Particulate Fallout Predictions for Clean Rooms" by Otto Hamburg in The Jour. of Environmental Sciences, May/June 1982 pp.15-20.
You can set out "witness plates" for various lengths of time and observe the amount of fallout you get and compare these results with environments of different load suspensions. Your witness plates should be identical to the substrates of interest....TEM grids and negatives, and be in the same place in your room. This can make a big difference. Particulate characteristics, air flow, humidity, substrates, etc. will be different from one person's environment to another's, meaning you cannot readily expect one person's experiences to be applicable to yours.
We have a particle prep lab that ranges between 1K-50K particles } = 0.5 micron per cubic foot, depending where you are and what phase the moon is in. We then have { Class 10 areas embedded within that where samples may be exposed to air. (Assuming that my laser counter's calibration is still good). Within the 20-50K areas that would be somewhat similar to yours, I would not expect to find significant dust accumulation on negatives overnight. However, I would guess that if the air was very dry, and there was plenty of air flow across the negatives, perhaps they would electrostatically collect enough by the next day to be a problem. I would not leave TEM grids out in this air if I was concerned about contamination.
Typical unfiltered office and home air where there is no smoke has about a few billion particles } = 0.5 microns per cubic foot. So if you have a rough idea of how rapidly dust accumulates there, you can apply the rules of 10 to get a VERY ROUGH idea of what you would get in the same environment if it had a few orders of magnitude less particulate. Again, I repeat, very rough.
I also have a story. I once worked in a cleanroom that had about 10,000 particles } = 0.5 micron per cubic foot in most areas (Class 10,000), and some Class 10 areas. About once every several days, I would find an enormous amount of dust on everything in the Class 10,000 areas and the clean room would have to be thoroughly cleaned. It appeared to be caused by "burps" in the plenum structure, caused by momentary pressure differentials, that allowed dust that had accumulated there to burp through seams in the ceiling. Solution: seal the seams. So your average dust suspension measurements may not reflect momentary, but significant, surges, for whatever reason.
Other useful references are:
FED-STD-209E Federal Standard for Clean Room and Work Station Requirements for Controlled Environments
MIL-STD 1246 Military Standard for Product Cleanliness Levels
Hope this helps.
Cynthia J. Zeissler Physical Scientist National Institute of Standards and Technology cynthia.zeissler-at-nist.gov 301-975-3910
A student will be assessing the penetration of various adhesive consolidants into egg tempera based paint layers. The consolidants would likely include animal glue, cellulosic ethers, and acrylics.
She has considered tagging the consolidant with one or more dyes to allow visual microscopic examination of depth penetration in samples cut in cross-section. Another means of assessing penetration would be micro-FTIR step-scan analysis of bulk cross-sections or thin-sections (1-5 microns).
Here's a question for the TEM folk on the list. Can anyone think of a feasible way to dope the consolidants with a metal(s) to allow mapping of depth penetration by SEM-EDS, TEM, or another technique?
I have only a limited knowledge of the use of metal-labelled antibodies to mark specific antigens using EM, and know this application I describe is quite different.
} Richard E. Edelmann, Ph.D. wrote: } } } } 4) CD-ROM storage: This makes alot of sense, since 99% of all } } computers come with CD-ROM readers. If you (or your users) have } } older CD-ROM readers, then they will need to cough up $80-150 to buy } } a newer one (10x IDE drive - 16x SCSI). } } } I don't get why users would have to buy a 10x IDE or 16x SCSI drive. Can't } the CD's be written in a standard format that can be read at any speed?? I } am about to buy a CD writer and apparently have missed something here. } TIA, Tom
I think Richard was saying that it may be worth the $100 or so necessary to upgrade the older 1X to 4X units to the faster speeds. I worked with a 1X for a few years. It was better than nothing, but it was slow!
Now a question for Richard. Do the tape drives really achieve 20 MB/sec or did you mean 20 MB/min? Our Ditto 2GB is on a flopy port and is doing well if it does 10 MB/min. ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-8216
Several of the members in the past have asked questions about Instrument rates and personal salaries. I recently saw a breakdown done on Mass Spectrometers. The breakdown was by three sectors - Industry, Government, and Universities. The data was presented for each instrument in each of the three sectors. The data was summarized including rates, the average age of the instruments and the staff to instrument ratio. Our lab was below the mean for most of the categories and now much effort is being place in trying to remedy the situation. With any luck we will acquire a new Mass Spec. and a staff member.
I was wondering if someone had a breakdown of this information handy on Microscopes and their support personal. I would like to see if we could do the same for the microscopy portion of the lab.
Thanks
Mike
=========================================================== Michael Dunlap lab (530) 752-0284 Facility For Advanced Instrumentation fax (530) 752-4412 University of California mrdunlap-at-ucdavis.edu Davis CA, 95616 http://carbon.ucdavis.edu ============================================================
We are in the process of preparing an article for the next "Focus on Microscopy" column for American Lab and would like your input. If you have 1. suggestions for image format or procedures for transporting images via the Internet or intranets, 2. tips and hints for using the inter/intranets for image libraries 3. tips and hints for communicating ideas on inter/intranets 4. microscopy or imaging Internet sites which you really enjoy please email me. If we can quote you, please add a line which gives us permission to use your name and your institution. If you have an extra minute or two, jot down a few comments about the type of work you do and what impact electronic media such as the Internet and/or intranets have had on your work.
Thanks! Barbara Foster Contributing editor American Laboratory
Ray Twesten says, in his response to Daraporn Arayasantiparb, that:
"...for a non-FEG TEM, the probe size is mainly determined by the demagnification of the finite source (i.e. the spot size selected). "
A few years ago I was involved in some work with a colleague where we made this assumption and generated some peculiar data. In the end we concluded that the probe size was, in fact, determined by the Cs of the objective lens and the convergence of the probe was too large. Depending upon where the collector aperture is placed, the convergence angle either increases or stays constant with increasing demagnification (i.e. smaller spot size settings). In our case, the probe size was in fact independant of the setting of the spot size control! (at least for the smaller sizes). It is essential to select an appropriate condenser aperture which matches the spot size setting, if that control is to have any meaning.
Tony Garratt-Reed
Anthony J. Garratt-Reed MIT Room 13-1027 77 Massachusetts Avenue Cambridge, MA 02139-4307 United States of America
Is there anyone out there that currently does particle size analysis by both Microscopy and Laser/Light Scattering techniques that can tell me if there is a listserver or newsgroup that deals with PSA specifically?
Dear all, Sorry if this has been discussed on the list before BUT.......
Does anyone know of a good method for Osmium Vapour staining/Osmium droplet staining for immuno-labelled grids? I am currently working on labelling some brain material from a honey bee, and have good signal, however I need to enhance the membranes a little more to accurately localise the labelling, than conventional staing with UA and Pb gives. Fixation is good and the tissue has been processed into Lowicryl K4M by PLT.
Like an inquisitive technician, I tried staining some Lowicryl sections of this tissue on some drops of OsO4 (with rinses afterwards) but came away with lots of tiny ppts all over the tissue! (non-specific ppts too! :-) )
So if someone could enlighten me with some trusty methods/references, that would be great!
Look forward to hearing from you,
Rich.
----------------------------------------------------------------------- Richard Lander Electron Microscope Technician South Campus Electron Microscope Unit Otago School of Medical Sciences P.O. Box 913 Dunedin New Zealand. Tel. National 03 479 7301 Fax. National 03 479 7254
"Southernmost EM Unit in the World!" ------------------------------------------------------------------------
Do any of you Reichert afficianados out there happen to know where= I can obtain a drive belt for an OmU2? I called Leica and they told= me parts are no longer available: This of course was after they= got over the shock of hearing one of these babies hasn't been turned= into a boat anchor yet. This little darling cuts like a hot knife= through butter, at least it did before the belt self-destructed= so I'd hate to see it go just because the belt is finished. If anyone= knows where I might be able to get a new belt, say for example when= you got that new Ultracut and left the OmU2 parts in that back drawer,= or make a replacement part of susbtitute goods (such as rubber bands,= duct tape, etc) I'd really like to hear from you.
Thanks a lot!
************************************************************ Everyone has a photographic memory. Some don't have film. ************************************************************ Laura Rhoads Electron Microscopy Facility Director Department of Biology Western Kentucky University 1 Big Red Way Bowling Green, KY 42101-3576
It is my experience that CDs written with our HP 4020 drive cannot be read on 1X CD readers. I have even encountered some 2X drives that don't like the written discs. (Discs are written at 1x speed.) We have 2 writers and the same holds true for CDs written by either drive. Never had a problem with a 4X or higher reader.
Bob Holthausen Pall Corporation Port Washington, NY
I think Richard was saying that it may be worth the $100 or so necessary to upgrade the older 1X to 4X units to the faster speeds. I worked with a 1X for a few years. It was better than nothing, but it was slow! Now a question for Richard. Do the tape drives really achieve 20 MB/sec or did you mean 20 MB/min? Our Ditto 2GB is on a flopy port and is doing well if it does 10 MB/min. ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-8216 E-Mail: wesaia-at-iastate.edu (or: wes-at-ameslab.gov) http://www.marl.iastate.edu/marl/ (re: SEM) http://www.public.iastate.edu/~iprt_info/cfce/ (re: coal) electron microscopy, x-ray analysis, image analysis, computer applications
wesaia-at-iastate.edu on 11/12/97 12:22:56 PM
To: Microscopy-at-Sparc5.Microscopy.Com cc: (bcc: Bob Holthausen/SLSNY/Pall/US)
At 08:56 AM 11/12/97 -0500, edelmare-at-casmail.muohio.edu wrote: } 3) Don't be so quick to reject tape storage. Tape storage can be } extremely cost effective in larger formats, speed can be a problem } but going back to older work-around solutions, users wishing to work } on files on Tuesday could transfer the data from tape to HD Monday } night, and then transfer back - alternately, utilizing a file server } for this task (File Server: HD's, Tape drives, CD-drives for } handleing just data storage: reading and writing only, no running of } other software) would free up workstations for computational work. } Yes, random access is limited but if you're looking at serial section } reconstruction you'll be accessing sequentially anyway. CON: You } have to a have a drive to read and write the data (unlike ubiquitous } CD-ROM readers). } } Some general tech specs: } } 2GB drives ($600), 11MB/sec, tapes -at- $10 [ $0.50/100MB] } } 2-4GB drives ($350-600), 29-42MB/sec, tapes -at- $31-10 } [$1.50-0.25/100MB] NOTE: Cheaper drives + More $ tapes } } 4-8GB drives ($750-1,200), 32-66MB/sec, Tapes -at- $16-28 [$0.40 } - 0.22/100 MB] } } 7-14GB drives ($700-1,600), 60-120MB/sec, tapes -at- $16 [$0.22 - } 0.11/100MB] } } 12-24GB drives ($1,000-1,300), 120-132MB/sec, tapes -at- $32-42 [$0.35 - } 0.13/100MB] } } Drives go upto the following: 20-40GB, 32-64GB, 70-150GB, 100-200GB, } 140-280GB -at- $2,000-9,000. } } NEW Iomega Ditto Max drives: upto 7GB* drives -at- $200, upto 10GB* } drives -at- $300, (*these are compressed data values, compression will } slow down transfer speeds) with transfer speeds upto 36MB/sec and } tapes running $20/3gb - $35/10GB [$0.20-$0.30/100MB].
I think you may have misquoted the above tape speeds. Based on my experience, it is more likely that the above speeds are in MB/minute instead of MB/second. That would place typical tape-drive speed more in line with that of the Zip-drive. Also, most tape-drive software accesses the tape in a sequential fashion, not randomly like the other media. This means when you wish to relocate a particular file, it will take a lot longer to get it from tape than from conventional media.
Jeff Ingeman Development Engineer Department of Anatomy & Neurobiology Department of Physiology & Biophysics University of California - Irvine (714) 824-7536 jingeman-at-uci.edu jingeman-at-aol.com jingeman-at-yahoo.com
Does anyone know if a TEM negative carrier is available for the Polaroid Sprintscan 45 Negative Scanner yet? Anyone having any experience with the carrier (if it exists) and this scanner, could you send me a little message on what you think of the system?
Thanks.
-Scott Walck
Walck-at-PPG.com
Scott D. Walck PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of S.D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
} Do any of you Reichert afficianados out there happen to know where I can } obtain a drive belt for an OmU2? I called Leica and they told me parts are } no longer available: This of course was after they got over the shock of } hearing one of these babies hasn't been turned into a boat anchor yet. } This little darling cuts like a hot knife through butter, at least it did } before the belt self-destructed so I'd hate to see it go just because the } belt is finished. If anyone knows where I might be able to get a new belt, } say for example when you got that new Ultracut and left the OmU2 parts in } that back drawer, or make a replacement part of susbtitute goods (such as } rubber bands, duct tape, etc) I'd really like to hear from you.
Hi Laura,
We also have a Reichert OmU2 that we loaned out to another department. It still is operating. We did replace the drive belt (and even the shock absorbing motor mounts) before we loaned the instrument out. (Yes, it worked fine after our repair.) If I recall properly, the drive belt from the motor to the microtome is really just a large O-ring and we found one at a truck bearings repair place. So, take the belt to your local hardware store or contact an O-ring supplier (I can give you the address if you need one). You should also be aware that there is an O-ring kit available to make any size O-ring by simply cutting the stock to desired length and gluding the ends together. They do work well.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
Hello Richard...we also had some problems with inadequate contrast of K4M embedded tissue which had been immunolabeled. Our solution was to stand the grids up on edge (in a slot cut into a wax sheet), to put the grids with the wax in a petri dish, then to add a few drops of OsO4 (2% aqueous) before covering the petri. We found that the osmium vapors imparted good contrast after about 30 minutes at room temperature.
Good luck!
Doug Keene Shriners Hospital for Children ---------------------- Doug Keene DRK-at-shcc.org
See Hayat & Miller, Negative Staining, McGraw-Hill, 1990.
Doane & Anderson, Electron Microscopy in Diagnostic Virology, Cambridge U Press, 1987.
Miller, J EM Technique (now Microsc Res & Tech) 4:265-301, 1986.
Tyrrell & Kapikian, Virus Infections of the Gastrointestinal Tract, Marcel Dekker, 1982.
I would NOT, I repeat ***NOT*** recommend fixing any virus sample before attaching it to the grid. If you are concerned with pathogenicity, you can fix it after allowing it to adhere to the grid, then wash with water and stain, or you can UV both sides of the grid after staining. For speed in reporting clinical results (we do almost 1000/year), we look at negative stains of potentially infectious material without fixing by keeping a separate specimen holder for "dirty' grids and another for nonpathogenic material such as sections. The grids are then UV'ed before storage.
Anything fixed by aldehydes, especially glutaraldehyde becomes less sticky after fixation. The most important reason that viruses in a clinical sample should not be fixed before gridding is that you decrease the numbers that stick to the support film. If you are close to the threshold of detection, you may decrease the numbers below that level.
Many of the described methods for fixing viruses before gridding include ultracentrifugation to concentrate them. This may be fine for samples such as stool from gastroenteritis patients where there are likely to be lots of virus, but for cerebrospinal fluid and other liquid samples, likely to have few viruses, you don't want to take the chance of losing any. If I were looking for an unknown virus in any sample, not knowing beforehand whether it were positive or negative, I would not want to lower my chances of finding something.
Further, adding fix to a dirty sample like stool can glue contaminants together. You can form large clumps that trap viruses and then are pelleted out in a low speed spin, effectively decreasing the number to see on your grid. Also, junk can be glued to viruses coating them so that they're unrecognizable.
It has been reported that some viruses have altered morphology after fixation. If you are looking for a novel virus, you should look at fixed and unfixed virus.
Misc. facts: We use uranyl acetate which fixes and preserves viral structure; it does NOT kill all viruses, but we use it because PTA is known to destroy some viruses, particularly reo- and rotaviruses. While you can observe them immediately after PTA staining, if you want to store them to show the prof or the medical student tomorrow or next week, you have to fix them and then wash with water before staining--adding more steps. Uranyl acetate is radioactive. PTA stains the spikes on viruses better (e.g., paramyxoviruses).
NOTE WELL: Finally, looking in stool for a virus that may be in brain may or may not yield positive results. If your question is "Does the patient who has viral encephalitis also shed virus in stool," then your search is valid. If your question is "Does the patient who has encephalitis have viral encephalitis," then you can't be sure a negative result from stool examination is valid. Not all viruses that cause viral encephalitis are shed in stool. Some never are; some are shed only for a short window of time. Some viruses that cause encephalitis could not be identified in stool, even if they were shed (e.g., alphaviruses, flaviviruses, bunyaviruses). Stool is just too dirty; there are too many membranes and vesicles that resemble viruses, and these viruses don't have a morphologically recognizable nucleocapsid. Finally some viruses cause a post infectious encephalomyelitis which appears to have an autoimmune component, and little or no virus is seen--even in brain. The only viruses you're really likely to see in feces of encephalitis patients are Picornaviruses (e.g., enteroviruses, Coxsackie viruses).
Feel free to call or email if you have questions.
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
} Date: Wed, 12 Nov 1997 15:46:14 -0600 } To: Microscopy-at-sparc5.microscopy.com } From: laura.rhoads-at-wku.edu (Laura Rhoads) } Subject: Drive Belt for OmU2
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hey there, } } Do any of you Reichert afficianados out there happen to know where I can } obtain a drive belt for an OmU2? I called Leica and they told me parts are no } longer available: This of course was after they got over the shock of hearing } one of these babies hasn't been turned into a boat anchor yet. This little } darling cuts like a hot knife through butter, at least it did before the belt } self-destructed so I'd hate to see it go just because the belt is finished. If } anyone knows where I might be able to get a new belt, say for example when you } got that new Ultracut and left the OmU2 parts in that back drawer, or make a } replacement part of susbtitute goods (such as rubber bands, duct tape, etc) } I'd really like to hear from you. } } Thanks a lot! } The material that is used for the drive on the Omu2 is quite different to the material that is used for O rings, and has quite different damping properties to the nitrile rubber oring cord. It can be successfully rejoined by melting the ends against a heated copper plate (or similar) and pressed together and then trimmed. We have also sucessfully used "superglue" after trimming the ends with a grease free razorblade. Very little length is lost. Similar material to the original is also supplied at places that specialize in Vbelt drives, and it too, is also normally joined by melting
Hello everyone ! We want to change our fixation and processing protocol for pulmonary tissues and cells to use HEPES buffer instead of sodium cacodylate buffer. Is anybody out there who has made her/his experience with HEPES buffer in conventional epoxy resin embedment. We are especially interested in experience of lipid retention/extraction.
Heinz Fehrenbach (Inst. of Pathology, TU Dresden, Germany)
1. Generally you use the gold on the mica and just stick the mica to the sample stub (in STM ensuring there is electrical contac between the gold and the stub). Then you generally image downwards finding large islands and flat areas of gold.
An alternative method, making larger areas of flat gold, is to remove the upper layers of gold from the mica - see " Uniformly flat gold surfaces: Imaging the domain structure of organic monolayers using scanning force microscopy" by Stamou_D, Gourdon_D, Liley_M, Burnham_NA, Kulik_A, Vogel_H, Duschl_C in LANGMUIR, 1997, Vol.13, No.9, pp.2425-2428 for a clearer description of this method.
2. I don't think there is.
3. Make it youself.
Giles Sanders Laboratory of Biophysics and Surface Analysis School of Pharmaceutical Sciences University of Nottingham
} Date: Thu, 13 Nov 1997 11:34:48 +0100 } To: ListServer-at-MSA.Microscopy.Com } From: Toufiq ELALLAM {elallam-at-lget.ups-tlse.fr} } Subject: financial support for postdoctoral } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We are using the BioRad confocal microscopes to collect stacks of 2-D pictures, transform these into 3-D using a Silicon Grapho=3DEDcs based syste= m with the VoxelWiew software. One of our interests is pathological conditions of the human brain, and we collect stacks os pictures from individual pyramidal or non-pyramidal neurons, visualized by intracellular injections of e.g. Lucifer Yellow, which spreads to every part of the neuron, including all spines. The neurons are collected with a presice location registered from an identified cortical (so far) region, dissected from either peroperative material or postmortem cases.
Characteristic morphological abberrations have been noted in cases of e.g. irretractible epilepsy, and Rett's syndrom. We are further looking to collect infantile autism cases and schizophrena cases.
We have in store many thousands of identified neurons with patient history and other data, which can be transferred to researchers in other parts of the world, for comparison with their own data, and for increasing case numbers to improve statistics. We have done succsssful transfers via the internet.
References:
Atypical pyramidal cells in epileptic human cortex: CFLS and 3-D reconstructions. P Belichenko, A Dahlstr=3DF6m, C von Essen, S Lindstr=3DF6m, C= Nordborg =3D and P Sourander NeuroReport, (1992) 3: 765-768
Application of confocal microscopy for the study of neuronal organization in human cortical areas after microinjection of lucifer yellow P. V. Belichenko, A. Dahlstr=3DF6m, and P. Sourander In: Biotechnology Applications of Microinjection, Microscopic Imagin= =3D g and Fluorescence. Ed. P.H. Bach et al. Plenum Press, New York,(1993) pp.29-35.
Dual channel confocal laser scanning microscopy of lucifer yellow- microinjected human brain cells combined with Texas red immunofluorescence P. Belichenko & A. Dahlstr=3DF6m. J. Neurosci. Meth.(1994) 52: 111-1= 18
Rett syndrome: 3-D confocal microscopy of cortical pyramidal dendrites and afferents Pavel V. Belichenko, Anders Oldfors, Bengt Hagberg, and Annica Dahlstr=3DF6m NeuroReport (1994) 5: 1509-1513
Dendritic morpholgy in epileptogenic cortex from TRPE patients, revealed by intracellular Lucifer Yellow microinjection and confocal laser scanning microscopy. Pavel V. Belichenko, Patrick Sourander, Kristina Malmgren, Claes Nordborg, Claes von Essen, Bertil Rydenhag, Sivert Lindstr=3DF6m, A= nd=3D ers Hedstr=3DF6m, Paul Uvebrant, Annica Dahlstr=3DF6m. Epilepsy Research (1994) 18: 233-247
Micromapping of the Human Brain: Three-Dimension Imaging of Immunofluorescence and Dendrictic Morphology Using Dual- Channel Confocal Laser Scanning Microscopy Pavel V. Belichenko and Annica Dahlstr=3DF6m Human Brain Mapping (1994) 1: 185-193
Morphological aberrations in therapy resistant partial epilepsy (TRPE); Confocal laser scanning and 3-D reconstructions of Lucifer yellow injected atypical pyramidal neurons in epileptic human cortex P Belichenko, P Sourander and A Dahlstr=3DF6m. Molec. Neurobiol.(1994) 9: 245-251
Studies on the 3-dimensional architecture of dendritic spines and varicosities in human cortex by confocal laser scanning microscopy = =3D and Lucifer Yellow microinjections. Pavel V. Belichenko , Annica Dahlstr=3DF6m J. Neurosci. Methods (1995) 57: 55-61
Contacts between serotoninergic fibres and dorsal horn spinocerebell= ar tract neurones in the cat and rat; a confocal microscopic study. Jankowska, DJ Maxwell, S Dolk, P Krutki, PV Belichenko and A Dahlstr= =3D =3DF6m. Neuroscience,(1995) 67: 477-487,
Mild cortical dysplasia: A three-dimensional study of dendritic morphology Pavel V. Belichenko and Annica Dahlstr=3DF6m Dysplasia of cerebral cortex and epilepsy , Eds. R. Guerrini et al Lippincott-Raven Philadelphia-New York (1995) pp.65-70,
Mapping of the human brain in normal and pathological situations: t= =3D he single cell and fiber level, employing lucifer yellow microinjection= =3D , carbocyanine dye tracing, immunoflourescence, and 3D confocal laser scanning microscopy reconstraction. Pavel V. Belichenko and Annica Dahlstr=3DF6m Neurosci. Protocols (1995), 95-050-03-01-30
Confocal laser scanning microscopy and 3-D reconstructions of neuronal structures in human brain cortex. Pavel B. Belichenko and Annica Dahlstr=3DF6m NeuroImage (1995) 2: 201-207
Calretinin-positive Cajal-Retzius cells persist in the adult human neocortex P.V.Belichenko, D.M. Vogt Weisenhorn, J. Myklossy and M.R. Celio Neuroreport (1995) 6, 1869-1874
Morphological study of neocortical areas in Rett syndrome. P V Belichenko, B Hagberg, A Dahlstr=3DF6m J Neuropathol (1997) 93: 50-61
Interested parties may contact us at this e-mail address.
Annica Dahlstr=3DF6m, MD, PhD and Pavel Belichenko, MD, PhD Prof., G=3DF6teborg University Prof. Brain Res. Institute, Moscow
Annica Dahlstr=3DF6m, MD, PhD Professor Inst. of Anatomy and Cell Biology Div of Neurobiology Medicinaregatan 3-5 S-413 90 G=3DF6teborg
I have been involved with the maintenance of electron microscopes for 33 years. Firstly as a service engineer and then through my own training organisation where we train both operators and service engineers. We run=
regular maintenance courses around the world when we get a good idea of which cleaning materials and solvents are available. =
I have been watching the discussions with interest to see if there were many holes in the cleaning explanations. That said may I toss in my few pennies (cents) worth?
TUNGSTEN Gun Systems
The cathode assembly should be cleaned every filament change, the anode every other change and the electron gun at least once a year.
Materials - Almost any metal polish may be used to clean electron gun components however it must not be LONG LIFE. Long life additives coat th= e cleaned item with a polymer that causes chaos in the electron gun. Look out for any indication on the bottle or tube that the manufacturer is claiming that you will not need to clean the metalwork so often after usi= ng their product!
Method - Almost more important than the cleaning efficiency is our abilit= y to completely remove the polishing media. So many service call outs are due to problems caused through inefficient removal of the media. For thi= s reason it makes sense to use a metal polish that is easily removed by a solvent for tungsten. In this way we not only remove the metal polish bu= t also clean the areas that are difficult to approach with the polish, nook= s and crannies! Also very important is the need to clean without damaging the component, scratching it or placing cotton hairs within the "traps" that the manufacturers seem to put in our way. The best cleaning techniq= ue is a wet clean, that is to use solutions and an ultrasonic cleaner. In this way the damage that mechanical forces apply to the components are minimised. Sure the cathode aperture may need a little more encouragemen= t to give up its deposit but only do this if the wet cleaning procedure fal= ls short. We like "Silvo" or "Bluebell" or "Brasso", liquid metal polishes that will mix with a dilute ammonia solution to form a cleaning media, b= ut a solution that may be removed with further washes in dilute ammonia. Th= e mix - 10% metal polish in 90% ammonia solution - where the solution is 1= 0% ammonium hydroxide in water. Place the components, one at a time, in the=
solution with their least important face down wards. Never put gun components together in the solution as they will damage each other. Do n= ot put an aluminium cathode in ammonia as it will go black, oxide! After 20=
minutes in an ultrasonic the component should be clean, wash off in runni= ng water and run for another 5 minutes in straight 10% ammonium hydroxide i= n water. Swill off with running water and then wash in alcohol and dry. =
NEVER throw away your solutions until you have reassembled the cathode as=
it is quite possible for the small screws to have fallen out and to resid= e in the debris at the base of one of the cleaning containers. If you do have a deposit remaining in the aperture area of the cathode a little mechanical effort with the cleaning media may be required,
The gun chamber IS important and this should be cleaned through disassemb= ly once a year, particularly with a TEM. Dirty guns hold gas and induce mic= ro discharge which spoils images. Clean the gun chamber with metal polish, remove the metal polish with dilute ammonia and buff up the walls with a clean chamois or dear skin leather. To retain the cleanlyness of the chamber, each time you change a filament buff up the walls with the leather. If the chamber smells, oily-ozone smell, but is not visibly stained, this is the result of discharge and all traces of the smell shou= ld be removed with dilute ammonia.
Look after your gun, it is probably the dirtiest area of the microscope, other than the specimen area in a SEM or the camera chamber in a TEM, its=
state will determine the ultimate performance of the instrument and your filament life.
LANTHANOM HEXABORIDE Technique developed by Biology E.M. Unit Canberra
Clean the cathode with 25% hydrochloric acid in water by immersing for 60=
seconds and then cleaning with a weak alkaline (ammonia or sodium hydroxide). Wash with water and then alcohol before drying.
LaB6 sources should last a long time (1000 hours plus) but they do need a= n intermediate cleaning session about every 250 to 350 hours. Some people amaze us by getting away with 1100 hours without cleaning but this is the=
exception not the rule.
Good luck!
*************************************************************************= ** ********************************************* Steve Chapman Senior Consultant
I received this inquiry today, and wondered if any of you could help. Please respond directly to Margaret, and not to the listserv.
Best regards, Steven Slap
} Return-Path: {nelsonm-at-Zeus.UCHSC.edu} } Delivered-To: ebsience-at-vgernet.net } From: "Margaret Nelson" {Margaret.Nelson-at-UCHSC.edu} } Organization: UCHSC Educational Support Services } To: ebs-at-ebsciences.com } Date: Tue, 11 Nov 1997 16:32:37 MST-0700 } Subject: Spurr Resin Toxicity } X-Confirm-Reading-To: "Margaret Nelson" {nelsonm-at-Zeus.UCHSC.edu} } X-pmrqc: 1 } Priority: normal } } At the suggestion of my doctor, I have been searching the internet } for information on the long-term effects of exposure to Spurr Resin. } From your web page, I was able to obtain the ingredients list, and } also a comment about a carcinogenic risk. This is the only } information I have found so far. } } Would you perhaps know of other sites I might try, or of physicians } or researchers who might know of others who have had an exposure to } this resin. I have not worked in the field of electron microscopy } for over 11 years, but the ramifications of the resin spill to my } legs (manifesting as spontaneous bruising) continues. I'm wondering } if others have experienced this, and what the future may hold. (What } sort of carcinogenic risk does this resin carry?) } } I'd be greatful if you could put me in touch with someone who might } have some useful information to share, and/or with a researcher } studying these questions who would be interested in my experiences. } Thanks for your time. } } Margaret G. Nelson } Univ. of Colo. Health Sciences Center } 4200 E. 9th Ave., Box A-066 } Denver, CO 80262 } (303) 315-6404, fax 315-6417 } } ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
I am an undergraduate and need help on staining blood cells which are fixed with 0.5% glutardialdehyde. My intention is to distinguish between monocytes and lymphocytes. Please do not tell me to use immunohistochemistry because it is not possible. The reason for this is that I also use very bright fluorescent latex beads ( FL-1 ) together with the cells and because of that I cannot take another fluorescent marker ( FL-2 ). I tried to stain my cells with Azur-B Eosin ( Romanowsky staining ) but it did not work. The cells just looked black or something like a very dark violet and I was unable to tell cytoplasm from nucleus. I think my problem is the glutardialdehyde but I have to use that!!! Does anyone have an idea about how to stain these fixed cells??????
Another solution could be that I separate lymphocytes and monocytes before I apply them for the microscope. Could anyone help me with that??? To isolate mnc from whole blood I use a Ficoll-Paque gradient centrifugation technique. I tied adhesion assays to separate monocytes from lymphocytes ( that means that the monocytes adhere to the petri dishes and the lymphocytes are mostly in the supernatant ), but the monocyte fraction is not very pure ( FACS analysis ) and I have a lot of debris because I have to use EDTA and a rubber policeman to get the monocytes off the plate.
Thank you very much!
Yours sincerely, Kathrin Augenstein
Institute of Immunobiology Stefan-Meier-Str. 8 79104 Freiburg Germany Fax. no.: 0041-761-2035446 e-mail: augenstk-at-ruf.uni-freiburg.de
10-Day Short Course on 3D Microscopy of Living Cells
June 17 - 28, 1998
and the Second, Post-course Workshop on
3D Image Processing June 30 - July 2
in association with the
UBC BioSciences Microscopy Facility
and the
Department of Computer Science
University of British Columbia Vancouver, BC, Canada
Organized by Prof. James Pawley University of Wisconsin-Madison
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 ten day residential course concentrating on all aspects of the 3D Microscopy of Living Cells will be again held at the University of British Columbia, in June of 1998. The course will cover every-thing from basic microscopy to the highest levels confocal microscopy.
The course will cover:
* Quantitative 2D light microscopy * 3D imaging in confocal and widefield * Fluorescent and backscattered light * Pixelation: The Nyquist Criterion * Lasers and laser tweezers * Objectives and aberrations * Scanning-systems: AODs and mirrors * Wide field/deconvolution techniques * Detectors: operation and performance * Optimal pinhole size/photon efficiency * Dye design, characteristics and use * How to keep your cells alive * Two-photon excitation * Video-rate confocal imaging * Measuring ion concentrations * Display and measurement of 3D data * Digital hard copy and storage
Morning lecture/demonstrations will lead to hands-on laboratory exercises each after-noon that will utilize most of the commercial instruments currently available for 3D micro-scopic imaging. Students will work in groups of 3 or 4 throughout the discussion and labo-ratory sessions, and will complete a live-cell 3D study on their own specimen.
Last year, 11 separate 3D microscopical workstations were available for student use under the supervision of an international faculty of 15. We expect to have even more workstations in 1998. Including manufacturers representatives, the teacher/ student ratio will be more than 1:1.
International Academic Faculty
* Jon Art University of Illinois * Pin Ching Cheng State U. of New York, Buffalo * Rachel Errington University of Nijmegen * Jim Pawley University of Wisconsin-Madison * Ernst Stelzer EMBL, Heidelberg * Michael Weis Agriculture Canada * Nick White Oxford University
International Commercial Faculty
* Dan Focht Bioptechs, PA * Ted Inou=E9 Universal Imaging, PA * Larry Keenan Cell Robotics, NM * Paul Millard Molecular Probes, OR * Sigrid Myrdal Multidimensional Imaging, WA * Paul Negulescu Aurora Biosciences, CA * Hans Van der Voort Scientific Volume Imaging, NL
TUITION
Course tuition is $1,950 US and includes lunches. On receipt of 50% deposit, all students will receive preliminary group assignments and a copy of the textbook, Handbook of Biological Confocal Microscopy, (Plenum, 1995). The tuition fee includes single tickets for the Opening Reception, the Manufacturer's Reception and the Beach Party, the textbook and all handouts. Accommodations and meals other than lunch are not included in the tuition fee. APPLICATIONS
Applicants will complete a questionnaire to assess knowledge level and field of interest. Enrollment will be 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. Application forms can be down-loaded from the WWW site at
Prof. James Pawley, Rm. 1235, 1500 Johnson Dr., Madison, WI, USA 53706. Phone: 1-608-263-3147, Fax 1-608-265-5315, Email: jbpawley-at-facstaff.wisc.edu
Application deadlines: Application forms must be received by March 1, 1998!
Successful applicants will be notified by April 1, and a deposit of 50% must be received by April 15, 1998 to reserve your position. In general, refunds of the deposit will only be possible if your position can be filled from the Waiting List. The remainder of the fees are due before registration.
DATES:
Applications must be received by Mar. 1/98 Deposit due Apr. 15/98 Registration 8:00 - 7:00 pm Wednesday, June 17/98 Last class will end with lunch Sun., June 28/98
WHO SHOULD ATTEND?
The course is designed for biological research scientists and advanced graduate students who use, or plan to apply 3D microscopy in studies involving living cells. No previous experience in advanced light microscopy is required but applicants will be asked how they plan to use 3D microscopy and to describe a short research project involving living cells that they plan to carry out during the course. Students with other interests in 3D light microscopy will be welcomed if space permits but usually the course is heavily oversubscribed.
Classes meet from 8:30 -12:00 and 1:00 - 6:00 with lecture-demonstrations in the morning and laboratory sessions in the afternoon. On average, only four topics will be covered in each morning session. There will be enough 3D microscopy setups to permit groups of 3-4 students to "learn-by-doing" during a carefully designed set of laboratory sessions. Lab handouts will include detailed questions to stimulate group discussions.
=46rom Sunday to Friday, facilities and supervision will be available until 11:00 pm, for those students who wish to work on their own projects.There
USE OF LEARNING GROUPS
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 in the evenings 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 LIVING SAMPLES
Students must contact the Course Organizer 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.
****************************************************************************= * 3D Image Processing Workshop June 30 - July 2
The course will cover 3D image processing for measurement and display. Enrollment is limited to those attending the 3D Micro-scopy course. Tuition : $700 US (lunch incl.) Live-cell Course
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 most 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 in a computer laboratory belonging to the Computer Sciences Department at the University of British Columbia which contains 27 SGI Indy workstaions and much of the other equipment needed for the measurement and display of 3D digital image data. Software from a variety of vendors serving the 3D microscopy market will be described, demonstrated and available for use.
Course Organizers * Nick White Oxford University * Hans Van der Voort Scientific Volume Imaging, NL
=46aculty * Pin Ching Cheng State U. of New York, Buffalo * Rachel Errington University of Nijmegen * Alain Fournier Computer Science, UBC * Sigrid Myrdal Seattle, WA
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 SGI work-stations. Students will "learn-by-doing" with two to a machine. Lab handouts will describe specific exercises to be performed on "canned" data sets.
Students also attending the 3D Microscopy Course, will be able to analyze, process and display the 3D data they have collected from their own specimens. Facilities and supervision will be available until 11:00 PM, for students to work on their own data.
Campus accommodations are student rooms or suites situated in the Walter Gage Residence located two blocks from the lecture-lab facilities and one block from the Student Union Building. The Union contains a large cafeteria, lounge, bank, etc. Many of the rooms in the Gage Residence have breath-taking views of the mountains of North and West Vancouver, and of the Pacific Ocean. A variety of accommodation types are available: $Cdn ($US)
- Single room w/shared washroom $30(23)
- Single room w/private bath $59(44)
- Double room (kitchenette, priv. bath, TV, phone, double bedroom plus separate sitting room) $85(63)
- Triple suite (twin bedroom and queen Murphy bed in sitting room, balcony,
kitchen, bath, TV, phone) $99(74)
(All fees are per night. Add 15% VAT. US rates are approximate and vary with exhange rate.)
Students are encouraged to bring friends or family members to enjoy the pristine beauty of the Vancouver area and the miles of lovely beaches that surround the campus. Arrangements can easily be made to extend your stay before or after the course.
MEALS
Lunches and morning and afternoon snacks are provided. Other meals can be purchased in the Union Cafeteria or any of a number of nearby restaurants. The Opening Reception and the Beach Party and the Manufacturer's Reception are included in the tuition fee. Additional tickets can be purchased for spouses and accompanying persons.
TRAVEL
Air:
Since the normalization of airfares between Canada and the US, fares to Vancouver from other parts of North America have been substantially reduced. The University campus is a 20-minute taxi ride from Vancouver International Airport. Students will receive a comprehensive selection of tourist information after their application has been accepted.
Bus:
=46or family members wishing to see the many sights, Vancouver has an excellent system of inexpensive and convenient public transportation.
Tours: Tours of Vancouver and environs can be arranged.
For more information & application forms, please contact the Course Organizer:
Prof. James Pawley, 1500 Johnson Dr., Madison, WI 53706. Phone: 1-608-263-3147/265-5315 fax. Email: jbpawley-at-facstaff.wisc.edu.
**************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 1235, Engineering Research Building, FAX 608-265-5315 1500 Engineering Dr., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU "A scientist is not one who can answer questions but one who can question answers." Theodore Schick Jr.,
} } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Is there anyone out there that currently does particle size analysis by } both Microscopy and Laser/Light Scattering techniques that can tell me if } there is a listserver or newsgroup that deals with PSA specifically? } } Thank you in advance! } } Hi, there,
Most recently I have done a lot of particle size analysis in the use of either TEM or OP. The particles actually are such sort of precipitate in metallic systems. Would you put more details in terms of what would you really want to know, especially for what kind of materials you are working for.
Peiyi Wang
Research Fellow Department of Engineering Materials University of Southampton Southampton SO17 1BJ UK
I switched to HEPES about 14 years ago for both my Paraformaldehyde/Glutaraldehyde buffer and for my osmium buffer. Safer, better for environment, and cheaper (I think). I noticed no difference compared to cacodylate (which I had previously used). A couple of years ago, I got into a discussion in which a colleague was touting PIPES as a better alternative. The logic is that HEPES has a pKa of 7.5. I use it at 7.4 for my fixes. Aldehyde fixes tend to acidify over time, therefore, since you start on the low side of the pKa and continue to fight a drop on that side, you have less buffering capacity. PIPES on the other hand, has a pKa of 6.8 so if you started at 7.4, you would have more buffering capacity (0.5 on both sides of the pKa). This is good logic but I didn't bother switching. Good luck.
-------------. } } Hello everyone ! } We want to change our fixation and processing protocol for pulmonary tissues } and cells to use HEPES buffer instead of sodium cacodylate buffer. Is anybody } out there who has made her/his experience with HEPES buffer in conventional } epoxy resin embedment. We are especially interested in experience of lipid } retention/extraction. } } Heinz Fehrenbach (Inst. of Pathology, TU Dresden, Germany)
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
You may be able to circumvent the effects of glutaraldehyde on the 'stainability' your cells with borohydride reduction:
"The pH dependence of borohydride as an aldehyde reductant" Bayliss, O.B. and C.W.M. Adams. Histochemical Journal 11:111-116, 1979.
Remember that borohydride solutions deteriorate VERY quickly, within minutes of preparation. Good luck!
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 e-mail: mcauliff-at-umdnj.edu ***************************************************************
I am not sure whether you have a flow cytometer or not, but a fluorescent microscope will work as well. You can stain your Monocytes with CD14 monoclonal antibody that is conjugated to either FITC FL-1) or PE (FL-2). Your lymphocytes can be stained with CD3 monoclonal antibody that is conjugated to either FITC or PE. You can use the two monoclonal antibodies together in which case the monoclonal antibodies will have to be conjugated to different fluorochromes, for example CD14-FITC combined with CD3-PE You can obtain the monoclonal antibody combination from Becton Dickinson Immunocytometry Systems, 2350 Qume Drive, San Jose, CA 95131-1807, Phone Number ( 800) 223-8226 .
In addition Becton Dickinson also sell the "TriTest" in which three different leukocyte specific monoclonal antibodies are combined together, CD45 is conjugated to a very bright red fluorochrome PerCP and monocytes and lymhocytes have different binding capacities for the CD45 monoclonal antibody. Note CD45 is Pan leukocyte monoclonal antibody that identifies all leukocytes.
FIXATIVE
Try using paraformaldehyde instead of glutaraldehyde.
I hope the above information is of help to you. Good Luck.
Bob Holthausen 11/13/97 11:52 AM
To: Sam Coker/SLSNY/Pall/US-at-Pall cc:
Yours sincerely, Kathrin Augenstein
Institute of Immunobiology Stefan-Meier-Str. 8 79104 Freiburg Germany Fax. no.: 0041-761-2035446 e-mail: augenstk-at-ruf.uni-freiburg.de
Yet another question from the unpredictable world of multi-user facilities. One of our users embeds samples in epoxy and polishes them to eliminate topographical variables. He requires both imaging and EDS, which precludes metal coating for conductivity.
He was wondering about the availability of epoxies containing conductive components, which might allow us to use high-vacuum SEM with secondary electron imaging. Does such a thing exist? Has anyone ever used a standard epoxy and added their own conductive "secret recipes" before polymerizing?
I'm aware that carbon coating is an option, as well as painting conductive stripes of colloidal carbon or silver to the edge of the embedded materials and down to the aluminum stub. The idea of a conductive epoxy is intriguing, though, for the time and mess-saving possibilities.
As usual, thanks in advance for the always helpful replies I get to these queries.
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
We have been experimenting with the use of HEPES as a non-toxic substitute for NaCacodylate for TEM during the past 6 months. So far the results have been good for mammalian and amphibian tissues (cardiac muscle, pancreas, heart, tongue). The membranes look good and we seem to have no trouble retaining lipid droplets in frog atrial muscle. I would be interested in anything anyone else has to offer. Also, I am assuming that HEPES is less toxic than Cacodylate. If anyone knows otherwise, I would like to hear about it.
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-131 University of Connecticut Storrs, CT 06269 Ph: 860-486-3588 Fax: 860-486-1936
Hi everybody Here is my (short)experience on CDR. I have a Sony CDR 928 on IDE Bus ( all my system is IDE not SCSI)and the first pb I encounter is that my old Windows95 version never never recognize this CDR. So I installed an OSR2 version on a new disk and all was OK. The software is easy CD pro from adaptec (upgraded for IDE). First writing (600Mo) no problem but impossible to read the datas on my 1 years old CD 8x. I check on many other even on a very old 2x Mitsumi and all was OK. So I change for a 24x Pionneer (the CDs were from Sony). All that takes at least one week late at night! My conclusions are the following: Writting speed doesn't affect reading, it's just a safety, the slower the safest. Never use Multisession, the price of CDs is now as low as 2$ and doesn't justify that. I use different manufacturer for CDs (SONY TRAXDATA...) without PB. You can never be sure that your work can be read on all CD for example I've made an Audio which is readable by all CD except on a JVC CD! I enclose a part of my Easy CD pro user's manual that prove that manufacturers know the problem. I have no interrest in any of the above company (just problem with some!) and this is my personnal opinion. If more information needed pls Email me.
FROM EASY CD PRO VERSION 2.0 ADAPTEC USER'S GUIDE
Problems Reading Recordable CDs If you have successfully written a CD but have problems reading it, there are a number of possible reasons : * If the CD can be read on the CD recorder but not on a standard CD-ROM drive, check in Disc Info and Tools to make sure that the session containing the data you just wrote is closed. CD-ROM drives cannot read data from a session which is not closed. =95 If your CD is ejected, or you receive an error message, or you have random problems accessing files from the CD, the problem may be that your CD-ROM drive is not well calibrated to read recordable CDs.
a If you recorded the CD using the DOS filenames option in the File Names tab, but there are nonetheless difficulties in reading back the CD on DOS or Windows OR 3.1 system, it may be that you have an older version of MSCDEX (before version 2.23) on your system.
Problems Reading Multisession CDs If you can see only data recorded in the first session on the CD but not in subsequent sessions, it may be that =95 You recorded the CD in CD-ROM (Mode 1) format, while your multisession CD-ROM drive only recognizes CD-ROM XA (Mode 2) multisession CDs.
or, =95 Your CD-ROM drive does not support muti session at all.
If you can see only data recorded in the last session, you may have forgotten to link your new data with data previously recorded on the CD. Make sure to select a track in the Load Contents tab before recording.
CD-ROM Drive Incompatibility with Recordable CDs Sometimes, it appears that you wrote a CD without trouble and can read it on your CD recorder ; however, when you put it in a standard CD-ROM drive, the CD is ejected, or you get error messages such as no CD-ROM or not ready reading, or you have random problems accessing some files or directories. You may find that the problems vanish completely when reading the CD on a different CD-ROM drive. This maybe due to compatibility problems with some CD-ROM drives, especially older ones, and recordable CDs. Some CD-ROM drives' lasers are not calibrated to read recordable CDs, whose surface is different from that of factory-pressed CDs. If your CD-ROM drive reads mass-produced (silver) CDs but not recordable CDs, check with the CD-ROM drive manufacture to determine whether this is the problem. In some cases , an upgrade is available which will solve the problem. The combination of CD brand and CD recorder can make a difference. Use CD media recommended by your CD recorder manufacturer.
I know this isn't strictly microscopy, but in light of the recent discussion about data storage, perhaps someome will be able to help.
We have an HP 1300T (1.3 GB, 650 MB on each side) magneto-optical drive that has failed us. It still reads and writes the disks okay, BUT ONLY when it accepts the disks. Normally (99+%), it tries three times to load the disk and then gives up.
We blew a lot of dust out of the unit and have opened it up and cleaned every place we could easily find (and a few more that we couldn't). Yet it still fails to accept cartridges.
HP has told us that the unit can only be replaced at a net cost of $650. I have also been told that encountering a failure after 2-1/2 years, with use a couple times a week over that period, is about par for the course. To say the least, that sounds quite short to me. I don't particularly wish to invest more money in such a short-lived option. (There was always the question of obsolescence but apparently there is also one of short working life.)
So the request- Does anyone out there have a drive that we could make some arrangements with to retrieve the data from our library of cartridges. We have about 10 GB of data to pull off. Then we would like to get away from the M-O drive and go to another media, probably CD-rewritable.
If perchance we could borrow someone's drive for a short period, I think we would be glad to leave you with our inventory of cartridges after we are done with the exercise. Any takers?
---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-8216
We used to use some copper-filled diallyl pthallate (sp?) to embed coal. It was a hot-preesed, thermosetting material from Beuhler or Leco. However, there was still a substantial fraction of the surface that was nonconductive. I don't recall if we could count on it providing a conducting path to ground. I think we C-coated most of our samples anyway.
At that time I was looking at S in coal and had little trouble from the C coating.
At 11:00 AM 11/13/97 -0700, you wrote: } Hi, } } Yet another question from the unpredictable world of multi-user facilities. } One of our users embeds samples in epoxy and polishes them to eliminate } topographical variables. He requires both imaging and EDS, which precludes } metal coating for conductivity. } } He was wondering about the availability of epoxies containing conductive } components, which might allow us to use high-vacuum SEM with secondary } electron imaging. Does such a thing exist? Has anyone ever used a } standard epoxy and added their own conductive "secret recipes" before } polymerizing? } } I'm aware that carbon coating is an option, as well as painting conductive } stripes of colloidal carbon or silver to the edge of the embedded materials } and down to the aluminum stub. The idea of a conductive epoxy is } intriguing, though, for the time and mess-saving possibilities. } } As usual, thanks in advance for the always helpful replies I get to these } queries. } } Randy Tindall ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-8216
I have a suggestion for the poor fellow who gets dry, cracked skin on his fingers in the winter: cut off your fingers. All of them. An ax should work finebut you could also use a power saw. Then you will never have to worry about dry cracked skin on your fingers. It will also be difficult for you to type, so I will probably get slightly fewer stupid e-mails about non-microscopy subjects.
Epoxy Technology, Inc. manufactures a complete spectrum of epoxies including at least four that are electrically conductive; all contain silver. I have not used any of them myself. BTW, their EPO-TEK 353ND is the same as Gatan's G-1, which I do use.
Call them at 800 227 2201 for a catalog.
Michael K. Cinibulk UES, Inc. Air Force Research Laboratory Materials and Manufacturing Directorate Wright-Patterson AFB, OH 45433-7817 937 255 9339 phone 937 656 4296 fax cinibumk-at-ml.wpafb.af.mil
There is a position open now for an electron microscopy specialist . Prepare ultra thin sections, and photogragh them using JEOL 1200EX. Darkroom, Adobe Illustrator/Photoshop.
Salary commensurate with experience.
Mail or fax resume and two letters of recommendation to:
Dr. Peter Sterling 123 Anatomy/Chemistry BLDG Department of Neuroscience University of Pennsylvania Philadelphia, PA 19104-6058
The matter of cleaning parts and components of vacuum systems is discussed in considerable detail on p. 69 - 74 of my book 'Vacuum Methods in Electron Microscopy' (Ashgate Pub. Co. Tel: 800-535-9544 ). Cleaning methods are extremely important in determining the pumpdown characteristics of vacuum systems, and the practical ultimate vacuum attainable in them, because of the overriding contribution of the outgassing phenomenon to these processes.
In particular, on p. 72 I cite a recommendation by Peter Sewell, of Lab-6 Inc. (a company that deals extensively with lanthanum hexaboride), that lanthanum hexaboride deposits can be effectively removed from Wehnelt cylinders by soaking them for about a minute in a solution consisting of one part by volumne of concentrated hydrochloric acid and four parts water, followed sequentially by thorough rinses with water, dilute ammonia, deionized water, and isopropyl alcohol, whereupon they are dried with a jet of clean hot air.
Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:313-763-4788; Ph:313-764-3321
} I received this inquiry today, and wondered if any of you could help. } Please respond directly to Margaret, and not to the listserv. } } } } At the suggestion of my doctor, I have been searching the internet } } for information on the long-term effects of exposure to Spurr Resin.
I'd like to see the responses to this query on the listserver, in addition to the direct response, please.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
Do you now if we have a newgroup on glass sciences? Currently I am editing some data on glass transition temperatures on some simple systems such as Al2O3-P2O5 and MgO-SiO2. If anyone have referrences on these glass transition temperatures or viscocity or know if we have a glass news group, please let me know.
Acme Conductive Adhesives (Division of Allied Products Corp. / New Haven, CT) used to sell a product called "E-Solder No. 3022," which is a conductive epoxy containing silver. It was intended as a substitute for solder in electronics repair when soldering isn't possible. I haven't used it for SEM. Hope this is helpful.
Scott Schwinge University of Washington Friday Harbor Labs 360-378-2165
} Yet another question from the unpredictable world of multi-user facilities. } One of our users embeds samples in epoxy and polishes them to eliminate } topographical variables. He requires both imaging and EDS, which precludes } metal coating for conductivity. } } He was wondering about the availability of epoxies containing conductive } components, which might allow us to use high-vacuum SEM with secondary } electron imaging. Does such a thing exist? Has anyone ever used a } standard epoxy and added their own conductive "secret recipes" before } polymerizing? } } I'm aware that carbon coating is an option, as well as painting conductive } stripes of colloidal carbon or silver to the edge of the embedded materials } and down to the aluminum stub. The idea of a conductive epoxy is } intriguing, though, for the time and mess-saving possibilities. } } As usual, thanks in advance for the always helpful replies I get to these } queries. } } } Randy Tindall } Electron Microscope Laboratory } Box 3EML } New Mexico State University } Las Cruces, NM 88003
Randy Tindall wrote: } He was wondering about the availability of epoxies containing conductiv= e } components, which might allow us to use high-vacuum SEM with secondary } electron imaging. Does such a thing exist? =
Allied High Tech Products has a Conductive Mounting Powder (order # 169-10005) with carbon particles. Allied High Tech (800) 950-9347 or (310) 635-2466
or
Electron Microscopy Sciences has a Conductive Cold Mount (# 50452-01) wit= h copper particles. EMS (800) 523-5874 or (215) 646-1566
"I have no commercial or financial interest in the companies stated above= , except within Mexico."
On Thu, 13 Nov 1997 JMARDINL-at-IMO.Intel.Com wrote:
} I have a suggestion for the poor fellow who gets dry, cracked skin on his } fingers in the winter: cut off your fingers. All of them. An ax should } work finebut you could also use a power saw. Then you will never have to } worry about dry cracked skin on your fingers. It will also be difficult } for you to type, so I will probably get slightly fewer stupid e-mails } about non-microscopy subjects.
Thank you for your kind suggestion, which I will add to the summary list of responses that I posted to the list. We can only hope the suggestion will be archived for future generations of microscopists.
I trust that _no one_ will feel the need to unnecessarily clutter the list with redundant thanks.
The suggestion was certainly novel, but I would be remiss in not pointing out (so to speak) the unexpected downside. When using my new voice recognition software, I find my lips chap more easily. I will, however, seek advice for this problem elsewhere.
*Received: from SpoolDir by IM-PW (Mercury 1.13); Thu, 13 Nov 97 22:19:47 -1800 *Return-path: {Microscopy-request-at-sparc5.microscopy.com} *Received: from Sparc5.Microscopy.Com by mailer.inmat.pw.edu.pl (Mercury 1.13); * Thu, 13 Nov 97 22:19:36 -1800 *Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id LAA05022 for dist-Microscopy; Thu, 13 Nov 1997 11:45:18 -0600 *Received: from bubba.NMSU.Edu (bubba.NMSU.Edu [128.123.3.39]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id LAA05019 for {microscopy-at-sparc5.microscopy.com} ; Thu, 13 Nov 1997 11:45:10 -0600 *Received: from NMSU.Edu by bubba.NMSU.Edu (8.8.7/NMSU) * id KAA14431; Thu, 13 Nov 1997 10:55:39 -0700 (MST) *Received: from nestor.NMSU.Edu by NMSU.Edu (8.8.4/NMSU-1.18) * id KAA17006; Thu, 13 Nov 1997 10:54:48 -0700 (MST) *Received: from Microspace.nmsu.edu (pc-hadams.NMSU.Edu [128.123.5.46]) by nestor.NMSU.Edu (8.8.6/8.7) with SMTP id KAA20404 for {microscopy-at-sparc5.microscopy.com} ; Thu, 13 Nov 1997 10:56:50 -0700 *Message-Id: {3.0.2.32.19971113110035.00692198-at-cnmailsvr.nmsu.edu} *X-Sender: rtindell-at-cnmailsvr.nmsu.edu *X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.2 (32) *Date: Thu, 13 Nov 1997 11:00:35 -0700 *To: microscopy-at-Sparc5.Microscopy.Com *From: Randy Tindall {rtindell-at-NMSU.Edu} *Subject: SEM/EDS/Conductive epoxies *Mime-Version: 1.0 *Content-Type: text/plain; charset="us-ascii"
Well, this is maybe not exactly an unswer which one may excpect, but it may give an idea how to deal with the problem. We are using old electric discharge machine and it requires from time to time the samples to be glued. So to make, the glue or epoxy conaductive some carbone powder is add.
Regards
Witold Zielinski Warsaw University of Technology Narbutta 85, O2-524 Warszawa Poland
May 21-23 and May 25-27, 1998 North Carolina State University Raleigh, North Carolina, USA
and
June 15-18, 1998 Danish Technological Institute Taastrup, Denmark
This highly regarded hands-on course taught by expert faculty has been presented annually for more than 15 years. It deals with all phases of quantitative and computer-assisted imaging from acquisition and processing through measurement and stereological interpretation. Attendees receive The Image Processing Handbook plus a CD-ROM containing images, algorithms (Photoshop-compatible for Mac and Windows) and an extensive on-line tutorial and course notes on stereology and statistical analysis. The course is appropriate for scientists, technicians and administrators using or intending to use these techniques. Attendees typically come from materials science, geology, biological and medical sciences, pharmaceuticals, food science, industrial quality control, remote sensing, and other disciplines. You are encouraged to bring your own images for the hands-on lab sessions.
For detailed information and registration contact Alice Warren, Dept. of Continuing and Professional Education, N. C. State University, Raleigh, NC 27695-7401, 919-515-4195, fax 919-515-7614, email: alice_warren-at-ncsu.edu
Information is available on-line at the following sites:
} I have a suggestion for the poor fellow who gets dry, cracked skin on his } fingers in the winter: cut off your fingers. All of them. An ax should work finebut you could also use a power saw. Then you will never have to worry about dry } cracked skin on your fingers. It will also be difficult for you to type, so I } will probably get slightly fewer stupid e-mails about non-microscopy subjects. ---------------------------------------------------------------------------- --------------------------------------
I assume the writer, who did not sign his name, is trying to be funny. Frankly I think these remarks are offensive and purile.
E-mails about safety and the side effects of chemicals used in microscopy are entirely "on-subject". There are many reagents used in microscopy which cause many workers a lot of problems and will continue to do so long after the exposure has ceased. Dry skin problems and dermatitis are often caused by exposure to microscopy laboratory reagents.
I share this problem of dry, cracking, bleeding and painful finger tips caused by exposure over many years to Lowicryl K4M, even though I used gloves. I no longer use the chemical but any adverse stimulus such as cold, drying or exposure dehydrating agents can cause a flare up of the condition.
Information on the best way to alleviate such problems from fellow sufferers and/or occupational health experts is useful and relevant. The list is not just for "techie-talk". It is about the subject as a whole, and that includes safety and health. If you don't want to read the e-mails use the delete key before opening, as I and many of us do, on many subjects on this list which don't interest or concern us.
Salzburg, 14th Nov. 1997 ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} I received this inquiry today, and wondered if any of you could help. } Please respond directly to Margaret, and not to the listserv. } } } } At the suggestion of my doctor, I have been searching the internet } } for information on the long-term effects of exposure to Spurr Resin.
Caroline Schooley WROTE: Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
"I'd like to see the responses to this query on the listserver, in addition to the direct response, please."
I would like to have this/these informations too (either via MSA-server or directly to my e-mail-box: W.Muss-at-lkasbg.gv.at) Thank you very much for your efforts!
Only an addition: the new (actual) EIDE Bus ("DMA33")has a throughput of 33MB/sec and is thus nearly as fast as UW-SCSI.
Richard E. Edelmann wrote: Things to keep in mind with these numbers (1) you'll never see these speeds in reality - they are all determined in ideal situations not real world usage, but they are comparable with each other; (2) Actual transfer rates will vary depending on the interface used (i.e. the max. throughputs for the interfaces are EIDE (1-10MB/sec) vs SCSI (5-10MB/sec) vs SCSI-Wide (20MB/Sec) vs SCSI-UltraWide (40MB/Sec) vs parallel ports ( {1 vs Fiber Coupling (a.k.a. Fire Wire, 100+MB/Sec), (3) System configuration will also effect this; i.e. what CPU, what BUS speed, how much memory (RAM) what other components are in the system, what operating system, etc. , (4)
At 08:56 12.11.97 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
In reply to a question on the staining of liposomes posted on Nov 03, I replied with the following on Nov 04:
} .......I have a couple of suggestions:
} 1] Apply your suspension to the Formvar surface of the support film rather } than the carbon surface. You may 'catch' more of the particles that way......etc. etc.
It only showed up in my mail box today - November 13 (I am assuming it appeared in everyone's box today also). Does anyone else experience delays like this in their postings?
Master Bond EP 75 is a graphite filled two part epoxy which is electrically conductive. It is rated at 50 ohm/cm, but I measure about 10k ohms/cm.
Metal particles cast and polished in this material show no charging when imaged at fast or slow rates in BSE or SE mode in my SEMQ. Nice to see the original colors of a material through the light optics. X-ray count rates seem OK.
It may be suitable for high vacuum since it contains no solvents. I can't melt a hole in the stuff with a 100 nA focused beam.
Adhesion is excellent. In fact, it sticks to silastic mold perimeters.
Mixing is a little fussy. 3 parts to 100. A perfect mix might get the 50 ohm/cm value. Shelf life is supposed to be less than a year and it's expensive.
The grain size of the carbon filler is relatively coarse. Colloidal graphite might be better.
This posting is definitely to help you people and not Master Bond, a company I did not find especially cooperative toward experimenters.
Master Bond is located in Hackensack, NJ. Phone 201 343 8983.
Knowing the specific resin would be a help. I have several methods for H&E staining and they are dependant on the type of plastic the tissue in embedded in.
-- Begin original message --
} Does anyone have a method for the H and & E staining of resin sections. I } would appreciate any methods or references } TIA Joan Clark } } }
-- End original message --
regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
If ever an intrinsically conductive polymer is developed which can be cast or hot pressed, I want to know. Conductive polymers are not uncommon these days, but not in a form useable for specimen mounting. Current conductive resins use tricks like adding iodine to the polymer chain to free-up some conductance electrons. ...A nice "tracer" too - unless you need to analyze for I. I have only found them in milled, "set" powder form.
I have used the silver epoxy and it does work. The Ag flakes are usually rather large, however, leaving lots of epoxy to charge at high mag. It can also be rather expensive to use for large "met" mounts. For economy, embed the specimen in a small amount of Ag/epoxy, mount in conventional mount, prepare, then carbon paint the remaining insulator.
The carbon loaded thermoset also works in a limited way. I have had problems with this material being difficult to clean. It is extremely difficult NOT to leave a nearly (optically) transparent film on a polished metal surface. The compounds I have tried *seem* to be slightly soluble in alcohols. If dried on the surface (often blowing w/N2 is not enough), it is back to the polishing wheel for removal.
Of the commercially available materials, I have had the best success with copper loaded hopt press resin. I used to get mine from Struers, but I understand they no longer make it. It does have the problem of charging resin islands as the structure is a network of Cu surrounding "islands" of resin. Still tough if you need to be near an edge. The bulk conductivith is excellent.
I understand there is a similar material using iron loading, but I have not tried that one....
I have also "brewed" my own conductive "cold-set" resin by adding a very rich loading of zinc DUST to acrilic/polyester type resins. The bulk conductivity is poor, but was sufficient to prevent charging in my application. BEWARE of zinc dust. It is reactive, like many metal powders/dusts. Be sure it will not cause problems (boom!, fire!, etc:) with a resin you may choose to combine with it!
Whenever possible, I just "paint" around the specimen using carbon paint. Using a 0000 sable brush, C paint thinned to an appropriate viscosity, and a 20x stereo optical scope, it is possible to "run" the paint right to the specimen edge. If you are lucky, surface tension and the mount/specimen discontinuity will cause the paint to stop at the interface. Don't have too many cups of coffee before attempting this!
Woody White, McDermott Technology, Inc. http://www.mtiresearch.com
Also woody.white-at-worldnet.att.net http://www.geocities.com/capecanaveral/3722
A good source of belts of different types (and lots of other useful items) is the company:
Small Parts, Inc. 6891 N.E. Third Ave PO Box 381966 Miami, FL 33238-1966 305-751-0856
You should be able to find their catalog in your local machine shop. They have round polycord belts that can be cut to length and melted together, timing belts, and viton and Buna-N o-rings.
Hope this helps, Louie
At 3:46 PM -0600 11/12/97, Laura Rhoads wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Louie Kerr Research and Education Support Coordinator Marine Biological Laboratory 7 MBL Street Woods Hole, MA 02543 508-289-7273 508-540-6902 (FAX)
Salzburg, 14th Nov.1997 Dear Joan, I saw your posting and from the "Conc:" I know you are dealing with = Epon-Araldite sections (hydrophobic resin) to be stained with H&E (which stains well = on=20 deparaffinized sections, but more/less no staining on hydrophobic resins = is achievable). I do not know for which purpose it should/must be H&E = (maybe I could=20 offer you alternative staining methods),=20 but eventually 2 methods maybe will be successful:
1st one I didn=B4t test as yet (but if o.k., it would be a short and = elegant way): (I=B4m not sure about the reactions on the araldite component of your = resin mixture)
prerequisite: resin sections should adhere very strongly to the slide: = be sure that=20 sections are "glued" properly by heat (approx. 100-140 degr. C) by means = of a hot plate without folds.
Try: - Oxidize the sections (e.g. 1 -2% KMnO4 in A.bidest) for 2-5 min - wash by jet-stream (A.bidest) thoroughly the sections now will exhibit brownish to brown appearance (KMnO) - Bleach by 0.5 to 1.0 % Oxalic acid (in a. bidest) for at least 3-5 = min, move/gentle agitate your slide at some time within this incub.time. The sections have to be totally decolorized, maybe bleach a minute=20 or so more to be sure, no KMnO is within your section. - Jet stream washing (or slide incubation in a coplin jar, change at = least 2-3 times) - you don=B4t have to let dry the sections - try to stain with H&E. Maybe there is a staining effect (hopefully; = maybe it depends on the polymerization quality of your sections, the = thickness, the concentration/time of oxidation (solution) etc.......). IF NOT: -you have to remove, at least in part, resin components of your = sections: - you have to use protocols like saturated Na-or K-MetOH (Na-or K-methylate) or=20 saturated Na-or K-EtOH (Na- or K-ethylate) (receipts of LANE and EUROPA, 1960ies or so.....100 years ago) that means: oversaturated solution of absolute methanol or ethanol by=20 adding NaOH- or KOH pastilles more than the solution product is. Let stand in a brown lab glass flask (plastic stopper!!) about 2-3 = days. The overlaying phase turns more and more to a brown color. Don=B4t = swurl up the solution when handling flask for removal of solute! BE CAREFUL in PIPETTING the SOLUTE because IT IS STRONGLY = CORROSIVE!! Place one to two drops of the "Etching solution" (Na-/K-methylate = or ethyl- ate) unto the sections and let "work" for 1-3 min (you have to = test; depends on the section thickness, polymerization quality, bla, bla....) After incubation (please be aware of the corrosive properties: = shield eyes, fingers, etc.) jet-wash with absolute MetOH or EtOH, respectively. Do this washing vigorously/thoroughly/extended (since highly = alkaline solu- tions stick a long time on a glass surface or elsewhere!) If your sections adhere until that step, you=B4ll be lucky! Best then would be jet-washing (or even incubation into a coplin = jar) by=20 say 70% or 50% MetOH or EtOH, then down into a. bidest. Let stand = a while and be sure about no alkaline etching solution has remained = on your=20 section or slide. Then try H&E. Maybe this will yield sufficient staining for the = purpose you intended. Would greatly appreciate receiving a short note on the background = of using H&E on your EPON/ARALDITE sections.
Hope this helps best regards
Dr. Wolfgang MUSS Department of Pathology, LKA EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
---------- Von: Joan Clark[SMTP:j.clark-at-zoology.unimelb.edu.au] Gesendet: Freitag, 14. November 1997 16:15 An: Microscopy-at-sparc5.microscopy.com Betreff: Q: H & E staining of epon araldite sections - help needed
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
Does anyone have a method for the H and & E staining of resin sections. = I would appreciate any methods or references TIA Joan Clark
Many thanks for the helpful replies after my recent inquiry about conductive epoxies for SEM specimen embedding. I am passing on the replies for other listmembers who may have an interest in this.
From Mark Darus:
} I use "PolyFast" available from Struers. The description on } the container states; Phenolic hot mounting resin with carbon filler } for edge retention and examination in SEM. } A code on the container is: FAPSA } 40100036 } 6062-5754 } Look in a Struers catalog for more information, or call them for one } at 1-888-Struers. }
From Matthew Libera:
} I had a similar problem when I was a grad student. I used conductive } epoxies from a company called Epotek in Billerica, Massachusetts. I } never had great success getting the conductive epoxy to cure properly, } however. Perhaps you will have more success. } } } From Phil Oshel:
} Various companies make silver-impregnated epoxy for making conductive } joins--such as gluing a new target onto an (old) Hummer sputter-coater } electrode. Don't know if this would work directly for embedding, but it } suggests that you could buy silver painting for mounting specimens that's } dissolved in acetone (EMS sells this, I think, others likely do also), and } use it in the embedding resin.
From Winton Cornell:
} what about simply mixing graphite into the epoxy as it's prepared? } } From Brian Demczyk:
} Yes, there most certainly are conductive epoxies (containing, for example, } silver). Check any of the EM supply houses. You might also want to check out a company called EPON-Tek, or something } of the like. } } From John Hunt:
} } Sure. Buehler and probably Struers, LECO etc. make conductive } material for mounts. The copper ones were removed from the market } some years ago but Al filled and Carbon filled ones are still } available, I believe. The powder is used in a hot hydraulic ram type } mold. The specimen is placed with the side of interest face down. The } mounts are usually inch or inch and a quarter diameter. The sample } is then ready for polishing. Coating is not necessary unless the } sampled is non-conductive in which case one might as well use epoxy. }
From Eunsung Park:
} I usually use a Ag-dispersed epoxy (from SPI) to embed small specimens } for both SEM and TEM work. Howver, it doesn't eliminate the necessity of } conductive coating since the epoxy contains non-conducting polymers. Another } problem is that the eopxy is not cheap (I fon't have the price list in handy). } It is surely worth to try, though. Good luck. }
From Warren Straszheim:
} We used to use some copper-filled diallyl pthallate (sp?) to embed coal. It } was a hot-preesed, thermosetting material from Beuhler or Leco. However, } there was still a substantial fraction of the surface that was } nonconductive. I don't recall if we could count on it providing a conducting } path to ground. I think we C-coated most of our samples anyway. } } At that time I was looking at S in coal and had little trouble from the C } coating. } } From Michael Cinibulk:
} } Epoxy Technology, Inc. manufactures a complete spectrum of epoxies } including at least four that are electrically conductive; all contain } silver. I have not used any of them myself. BTW, their EPO-TEK 353ND is } the same as Gatan's G-1, which I do use. } } Call them at 800 227 2201 for a catalog. } } From Scott Schwinge:
} Acme Conductive Adhesives (Division of Allied Products Corp. / New Haven, } CT) used to sell a product called "E-Solder No. 3022," which is a } conductive epoxy containing silver. It was intended as a substitute for } solder in electronics repair when soldering isn't possible. I haven't } used it for SEM. Hope this is helpful. }
From Hermann Reese:
} Allied High Tech Products has a Conductive Mounting Powder (order # } 169-10005) with carbon particles. } Allied High Tech (800) 950-9347 or (310) 635-2466 } } or } } Electron Microscopy Sciences has a Conductive Cold Mount (# 50452-01) with } copper particles. } EMS (800) 523-5874 or (215) 646-1566 } } } "I have no commercial or financial interest in the companies stated above, } except within Mexico." } } From Witold Zielinski:
} Well, this is maybe not exactly an unswer which one may excpect, } but it may give an idea how to deal with the problem. } We are using old electric discharge machine and it requires from } time to time the samples to be glued. So to make, the glue or } epoxy conaductive some carbone powder is add. } }
This is what I have received to date. Thanks to all and my apologies if I missed anybody.
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
Only one addition to complete: the actual EIDE Bus ("DMA33")has a throughput of 33MB/sec and is thus nearly as fast as UW-SCSI and also has many of the features before only given by SCSI (less processor-power usage etc.)
Richard E. Edelmann wrote: Things to keep in mind with these numbers (1) you'll never see these speeds in reality - they are all determined in ideal situations not real world usage, but they are comparable with each other; (2) Actual transfer rates will vary depending on the interface used (i.e. the max. throughputs for the interfaces are EIDE (1-10MB/sec) vs SCSI (5-10MB/sec) vs SCSI-Wide (20MB/Sec) vs SCSI-UltraWide (40MB/Sec) vs parallel ports ( {1 vs Fiber Coupling (a.k.a. Fire Wire, 100+MB/Sec), (3) System configuration will also effect this; i.e. what CPU, what BUS speed, how much memory (RAM) what other components are in the system, what operating system, etc. , (4)
Dr. Arthur Schuessler University of Heidelberg Zellenlehre D-69120 Heidelberg Germany
since we have a serious problem with immunofluorescence, I hope somebody can help us. We used 0.5 and 1 micron sections of Unicryl embedded plant tissues for immunofluorescence. Sections were cut with an diamond knife / ultracut on water. We did labelings with second Antibodies (anti-rabbit) conjugated to FITC or Cy3. Once we had nice pictures, but 10 times we had --} } } } horrible spots ("stars") as background all over the regions the incubation drop was located, and very weak labeling. Also there are fluorescing droplets when using Cy3, I wonder if it is not ok. However, the FITC conjugated also shows the "stars". It really looks very bad - all is full of "dirt".
What could be the reason? In controls without first antibody the sections are always clean. We use TBS with 0.1 or 1% Casein, polylysin coated or uncoated coverslips, heat dried or not heated etc. With no first antibody always no stars. Is the antigen floating around?
Thanks a lot if there are any ideas ... Arthur Dr. Arthur Schuessler University of Heidelberg Zellenlehre D-69120 Heidelberg Germany
We are trying to evaluate a number of dimpler grinders for both plan view and cross-section TEM specimens and would like the opinion of people who might have used the models put out by the following 2 companies: VCR Group (Model 500i) and EA Fischione (Model 2000). These can be compared with the Gatan model (656) if the person has experience with those as well.
--
---------------------------------------------------------------- Mukul KUMAR, PhD Dept. of Mechanical Engineering Phone: (410) 516-8284 The Johns Hopkins University Fax: (410) 516-4316 3400, N. Charles St. E-Mail: mukul-at-jhu.edu Baltimore, MD 21218-2686 ----------------------------------------------------------------
Dear Lewis, } } I need a positive charge on formvar and or carbon coated copper grids. Any } suggestions or references would be appreciated. } Poly-l-lysine should do the job (unless you are going to apply a *very* high pH specimen). Just put a few microliters of 0.1% aqueous solution on the grid and air-dry. Good luck. Yours, Bill Tivol
Greetings, FWIW, I use either isopropyl based colloidal graphite or silver paint mixed 1:1 as a slurry with Duco household cement to affix samples to SEM stubs. It's not epoxy but it dries quickly, provides good conductivity and is easily scraped off to re-use stubs.
cheers ed
Edward J. Basgall, PhD The Pennsylvania State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618 http://www.personal.psu.edu/ejb11/ Privilege does not absolve one of ecological responsibility.
I agree with Stephen Griffiths reply and would like to see this thread kept open a little longer. Most of the biological types have acquired some sort of allergy or dermatitis along the way if they have been in the field for any period of time, and those who haven't need to be reminded that the three most important assets a microscopist has is their brain, eyes, and hands. Without all of these, you can no longer ply your trade. The "chapped hand" syndrome is not trivial, the skin is a line of defense for chemical and for pathogens, once broken, both are free to penetrate. I would reinforce use of proper gloving as a way to avoid major problems in the future for those whose hands haven't gone by the way. In addition, I would like to remind that we are liable for what happens to our subordinates and severe dermatitis can, in fact, be a handicap if it limits range of motion. Like most, Lowicryl started my dermatitis, although I noticed the effects within months of first starting to use it. Little seems to help. Superglue and Eucerin are my mainstays. I would like to thank James Martin for asking the question as well as those who responded. I have been looking over the pharmacy shelves, picking up some of the suggested remedies to try.
Dear James, I have experience with penetration of glues, preservatives, etc. into wood, and the best label was bromide. This shows up well in EDX, is completely soluble and you can brominate most organic compounds quite well. We also used it the trace the movement of epoxy resin in prepreg layup parts (composite materials). Brominate the consolidant, then trace the presence of bromine with an EDX linescan or careful P/B analysis of bromine. It works equally well in TEM for individual wood cell wall layers (brominate the lignin) or SEM for overall penetration depths. You wrote: } A student will be assessing the penetration of various adhesive } consolidants into egg tempera based paint layers. The consolidants would } likely include animal glue, cellulosic ethers, and acrylics. } } She has considered tagging the consolidant with one or more dyes to allow } visual microscopic examination of depth penetration in samples cut in } cross-section. Another means of assessing penetration would be micro-FTIR } step-scan analysis of bulk cross-sections or thin-sections (1-5 microns). } } Here's a question for the TEM folk on the list. Can anyone think of a } feasible way to dope the consolidants with a metal(s) to allow mapping of } depth penetration by SEM-EDS, TEM, or another technique? } } I have only a limited knowledge of the use of metal-labelled antibodies to } mark specific antigens using EM, and know this application I describe is } quite different. } } Thanks for your assistance with this inquiry. } } James Martin Regards, 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 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
Dear Randy, I have used the conductive cold-curing resin: "Technovit 5000" for several years, but the last time I tried, my usual suppliers no longer carried it. Electron Beam Sciences Inc. informed me that they have it available, when I asked on the listserver. Several other suppliers of mounting media have hot-press, conductive mounting media (Leco, etc.). For my usual metallurgical mounts, I mount in normal epoxy, poloish, then paint all of the top epoxy surface with carbon paint, overlapping the metal slightly. Then run a stripe down the side to the stub to connect. I use the carbon evaporative coating if I want to look at the very edge of the sample or if the sample is not conductive, and only use the conductive resin if the sample cannot be coated and the edge is important. You wrote: } } Hi, } } Yet another question from the unpredictable world of multi-user facilities. } One of our users embeds samples in epoxy and polishes them to eliminate } topographical variables. He requires both imaging and EDS, which precludes } metal coating for conductivity. } } He was wondering about the availability of epoxies containing conductive } components, which might allow us to use high-vacuum SEM with secondary } electron imaging. Does such a thing exist? Has anyone ever used a } standard epoxy and added their own conductive "secret recipes" before } polymerizing? } } I'm aware that carbon coating is an option, as well as painting conductive } stripes of colloidal carbon or silver to the edge of the embedded materials } and down to the aluminum stub. The idea of a conductive epoxy is } intriguing, though, for the time and mess-saving possibilities. } } As usual, thanks in advance for the always helpful replies I get to these } queries. } } } Randy Tindall } Electron Microscope Laboratory } Box 3EML } New Mexico State University } Las Cruces, NM 88003 } Regards,
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 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
Dear Listserver Readers, I note with interest that a competitor (Evex Analytical) has registered the following URL: http://www.thomson-scientific.com/ for their own use.
As a director of Thomson Scientific Instruments Pty Ltd I wish to state quite categorically that this company is in no way associated with us. Our website is in fact located at: http://www.werple.net.au/~tsi/
I shall leave it up to readers to make their own judgment regarding the ethics of Evex Analyticals action.
Thank you,
Paul Thomson Technical Director Thomson Scientific Instruments http://werple.net.au/~tsi/
We have a Zeiss 902 filter scope for sale. 6 years old. In excellent condition. Asking 28K. Will also deliver to your location. Presently on West coast. Call 732- 370-8082 for info.
What is the actual formula Sorensen used to make Sorensen's phosphate buffer? Not what you think it is, but What Dr. Sorensen stated in his paper in 1909, and what many of us have or thought we have been referencing for the past 88 years.
Some references say (Hayat) to use sodium phosphate and Potassium phosphate
Most other references state Sorensens Phosphate buffer has being made with sodium monobasic and sodium dibasic salts.
If anyone has the original reference to sorensens (biochem, 22, 253, 1909) on hand, I would be most appreciative if they would be kind enough to email or fax it to me.
Thanks in advance Gregory Argentieri Novartis Pharmaceuticals Corp 59 Rt 10 East Hanover, NJ 07936
Master Bond EP 75 is a graphite filled room temp setting, two part epoxy which is electrically conductive. It is rated at 50 ohm/cm, but I measure about 10k ohms/cm.
Metal particles cast and polished in this material show no charging when imaged at fast or slow rates in BSE or SE mode in my SEMQ. Nice to see the original colors of a material through the light optics. X-ray count rates seem OK.
It may be suitable for high vacuum since it contains no solvents. I can't volatilize a hole in the stuff even with a 100 nA focused beam.
Adhesion is excellent. In fact, it sticks to silastic mold perimeters.
Mixing is a little fussy. 3 parts to 100. A perfect mix might get the 50 ohm/cm value. Shelf life is supposed to be less than a year and it's expensive.
The grain size of the carbon filler is relatively coarse. Colloidal graphite might work better.
Master Bond is located in Hackensack, NJ. Phone 201 343 8983.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Question/debate of the day:
What is the actual formula Sorensen used to make Sorensen's phosphate buffer? Not what you think it is, but What Dr. Sorensen stated in his paper in 1909, and what many of us have or thought we have been referencing for the past 88 years.
Some references say (Hayat) to use sodium phosphate and Potassium phosphate
Most other references state Sorensens Phosphate buffer has being made with sodium monobasic and sodium dibasic salts.
If anyone has the original reference to sorensens (biochem, 22, 253, 1909) on hand, I would be most appreciative if they would be kind enough to email or fax it to me.
Thanks in advance Gregory Argentieri Novartis Pharmaceuticals Corp 59 Rt 10 East Hanover, NJ 07936
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Master Bond EP 75 is a graphite filled room temp setting, two part epoxy which is electrically conductive. It is rated at 50 ohm/cm, but I measure about 10k ohms/cm.
Metal particles cast and polished in this material show no charging when imaged at fast or slow rates in BSE or SE mode in my SEMQ. Nice to see the original colors of a material through the light optics. X-ray count rates seem OK.
It may be suitable for high vacuum since it contains no solvents. I can't volatilize a hole in the stuff even with a 100 nA focused beam.
Adhesion is excellent. In fact, it sticks to silastic mold perimeters.
Mixing is a little fussy. 3 parts to 100. A perfect mix might get the 50 ohm/cm value. Shelf life is supposed to be less than a year and it's expensive.
The grain size of the carbon filler is relatively coarse. Colloidal graphite might work better.
Master Bond is located in Hackensack, NJ. Phone 201 343 8983.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Question/debate of the day:
What is the actual formula Sorensen used to make Sorensen's phosphate buffer? Not what you think it is, but What Dr. Sorensen stated in his paper in 1909, and what many of us have or thought we have been referencing for the past 88 years.
Some references say (Hayat) to use sodium phosphate and Potassium phosphate
Most other references state Sorensens Phosphate buffer has being made with sodium monobasic and sodium dibasic salts.
If anyone has the original reference to sorensens (biochem, 22, 253, 1909) on hand, I would be most appreciative if they would be kind enough to email or fax it to me.
Thanks in advance Gregory Argentieri Novartis Pharmaceuticals Corp 59 Rt 10 East Hanover, NJ 07936
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Master Bond EP 75 is a graphite filled room temp setting, two part epoxy which is electrically conductive. It is rated at 50 ohm/cm, but I measure about 10k ohms/cm.
Metal particles cast and polished in this material show no charging when imaged at fast or slow rates in BSE or SE mode in my SEMQ. Nice to see the original colors of a material through the light optics. X-ray count rates seem OK.
It may be suitable for high vacuum since it contains no solvents. I can't volatilize a hole in the stuff even with a 100 nA focused beam.
Adhesion is excellent. In fact, it sticks to silastic mold perimeters.
Mixing is a little fussy. 3 parts to 100. A perfect mix might get the 50 ohm/cm value. Shelf life is supposed to be less than a year and it's expensive.
The grain size of the carbon filler is relatively coarse. Colloidal graphite might work better.
Master Bond is located in Hackensack, NJ. Phone 201 343 8983.
Yes I see the bouncing message. It is all coming from one host and I am attempting to contact the system adminstrator there a bit hard on a Sunday Morning.
I have temporairly disabled all mail to subscribers at that site and I hope that that will mitigate the problem in the short term.
After reading Paul Thomson's posting we have discovered that Evex Analytical is also using domain name www.mektech.com. Needless to say that Mektech Inc. was NOT, is NOT, nor will it ever be associated in any way with Evex Analytical. Mektech's true website is located at http://www.visionol.net/~mektech.
It is very regrettable that deception has been chosen as a tactic by an unscrupulous company claiming to serve the scientific community.
Mektech Inc. http://www.visionol.net/~mektech
-------------------------------- Paul Thomson wrote:
} Dear Listserver Readers, } I note with interest that a competitor (Evex Analytical) has registered } the following URL: http://www.thomson-scientific.com/ for their own use.
} As a director of Thomson Scientific Instruments Pty Ltd I wish to state } quite categorically that this company is in no way associated with us. Our } website is in fact located at: http://www.werple.net.au/~tsi/
} I shall leave it up to readers to make their own judgment regarding the } ethics of Evex Analyticals action.
Thomson Scientific is a wholly owned Research & Development subsidiary of Evex Analytical Inc.,
----------
Dear Listserver Readers, I note with interest that a competitor (Evex Analytical) has registered the following URL: http://www.thomson-scientific.com/ for their own use.
As a director of Thomson Scientific Instruments Pty Ltd I wish to state quite categorically that this company is in no way associated with us. Our website is in fact located at: http://www.werple.net.au/~tsi/
I shall leave it up to readers to make their own judgment regarding the ethics of Evex Analyticals action.
Thank you,
Paul Thomson Technical Director Thomson Scientific Instruments http://werple.net.au/~tsi/
At 14:15 14/11/97 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
"The mounting of Macromolecules for Electron Microscopy with particular reference to surface phenomena and the treatment of support films by glow discharge"
Jacques DuBochet, Michael Groom, & Shirley Mueller-Neuteboom. 1982
in
Advances in Optical and Electron Microscopy Vol. 8
Barer & Cosslet eds. Academic Press
Mel Dickson President, Australian Society for Electron Microscopy Director, Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
You know, there *is* some sort of legal redress for this. I'm not sure what it is, since I really didn't care about my own name (Yes, believe it or not, some guy in British Columbia bought up all the names of people who posted to a newsgroup I frequent. The domain "oliver.net" is owned, along with about 500 others, by a firm in Vancouver which markets email forwarding, I think).
If you are interested in looking further to see if others have your name, look at the internic site:
http://rs.internic.net/rs-internic.html and use the WhoIs function.
Here are some good sites re legal issues:
http://www.pitt.edu/~lawrev/58-4/articles/domain.htm (from the Pittsburgh Law Review) http://www.mindspring.com/~iprop/webdoc1.htm (a review of the NSI conflict resolution policy) http://rs.internic.net/domain-info/internic-domain-6.txt (the NSI document)
The Law Review Article is great. Here's an excerpt for the section on "squatters."
1. Squatters - Dilution to the "Rescue"
All the written decisions involving squatters involve one individual, Dennis Toeppen, who registered numerous trademarks of others as domain names, including "americanstandard.com," "panavision.com," "aircanada.com," and "yankeestadium.com."(367) In Panavision, the defendant registered plaintiff's trademark "Panavision" as a domain name, and used his WWW site to display an "aerial view[] of Pana, Illinois."(368) Toeppen demanded $13,000 to discontinue use of the domain name.(369)
The court found that Toeppen had violated state and federal anti-dilution statutes and ordered a preliminary injunction.(370) The marks, having been used since 1954, were in the eyes of the general public and producers, directors and movie studios, "famous" marks within the meaning of the act.(371) Toeppen's use of the domain name lessened the capacity of the mark to " 'identify and distinguish goods or services' " under the Dilution Act, and further diluted the mark by preventing Panavision from using its trademark verbatim as a domain name.(372)
...
and "parasites"
2. Parasites
a. Use of the Same Name Between Competitors - Infringement
Parasites who obtain a domain name corresponding to the trademark of a competitor will rightly be found liable for infringement.(390) In Comp Examiner Agency, Inc. v. Juris, Inc.,(391) the Central District of California granted a preliminary injunction against a defendant who used plaintiff's federally registered trademark as a domain name to "sell, distribute, advertise, and/or market its goods and services to Juris' target market of lawyers and law firms."(392) Although the court provided no explanation, its finding of trademark infringement under sections 32(1) and 43(a) of the Lanham Act is reasonable: the use of A's registered trademark by B to market the same products as A, to A's target market, is the paradigm of infringement and "passing off." The court enjoined the defendant from using "Juris" or any confusing variant; nonetheless, defendant was granted three months to continue using the domain name and web site to post a text-only referral notice to defendant's new site.(393)
On Sun, 16 Nov 1997, Mektech Inc. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } Dear readers, } } After reading Paul Thomson's posting we have discovered that Evex Analytical } is also using domain name www.mektech.com. Needless to say that Mektech } Inc. was NOT, is NOT, nor will it ever be associated in } any way with Evex Analytical. Mektech's true website is located at } http://www.visionol.net/~mektech. } } It is very regrettable that deception has been chosen as a tactic by an } unscrupulous company claiming } to serve the scientific community. } } } Mektech Inc. } http://www.visionol.net/~mektech } } -------------------------------- } Paul Thomson wrote: } } } Dear Listserver Readers, } } I note with interest that a competitor (Evex Analytical) has registered } } the following URL: http://www.thomson-scientific.com/ for their own use. } } } As a director of Thomson Scientific Instruments Pty Ltd I wish to state } } quite categorically that this company is in no way associated with us. Our } } website is in fact located at: http://www.werple.net.au/~tsi/ } } } I shall leave it up to readers to make their own judgment regarding the } } ethics of Evex Analyticals action. } }
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Thomson Scientific is a wholly owned Research & Development subsidiary of Evex Analytical Inc., }
Interesting. No doubt Mektech is one also. How many of your "wholly-owned subsidiaries" "just happen" to be the names of competitors?
Just curious, of course. I'm sure your ethical stance is spotless.
Well, I really don't usually follow up on *myself*, but I went ahead and did a WhoIs on EVEX. It turns out that EVEX, happens to have registered the following names:
I would be interested in the going rates for typical TEM charges , from reception of the specimen to the generation of prints, irregardless of interpretation. Has anyone accurate costing information on this?
Best regards, Jerome Jasso Children's Hospital Medical Center of Akron (330) 379-8279
I wish to thank those three people who answered my request which I repeat below. All three pointed me to suppliers in the U.S. (which is a bit expensive due to postage) or to standard EM-suppliers but there I have only found boxes for biological standard size thin sections (the 76 mm long ones). I am really surprised that this German Company should have been the only one to provide boxes for 28x48 mm size. Or are there so few Europeans on the list??
} } Hello everybody, } a question to the mineralogists on the list: } as the German provider (Krantz) for storage boxes for mineralogical } standard thin sections cannot provide them any longer I am looking for some } other source, preferably in Europe. } What I call "standard thin sections" are 28 x 48 mm sized. } Thank you in advance } Hiltrud } } Dr. Hiltrud Mueller-Sigmund } Institut fuer Mineralogie, Petrologie und Geochemie } Albertstrasse 23b, 79104 Freiburg (Germany) } Tel.: (+49)-203-6388/-6396 Fax: -6407 } } } hiltrud-at-ruf.uni-freiburg.de (Hiltrud Mueller-Sigmund)
Dr. Hiltrud Mueller-Sigmund Institut fuer Mineralogie, Petrologie und Geochemie Albertstrasse 23b, 79104 Freiburg (Germany) Tel.: (+49)-203-6388/-6396 Fax: -6407
I wish to thank those three people who answered my request which I repeat below. All three pointed me to suppliers in the U.S. (which is a bit expensive due to postage) or to standard EM-suppliers but there I have only found boxes for biological standard size thin sections (the 76 mm long ones). I am really surprised that this German Company should have been the only one to provide boxes for 28x48 mm size. Or are there so few Europeans on the list??
} } Hello everybody, } a question to the mineralogists on the list: } as the German provider (Krantz) for storage boxes for mineralogical } standard thin sections cannot provide them any longer I am looking for some } other source, preferably in Europe. } What I call "standard thin sections" are 28 x 48 mm sized. } Thank you in advance } Hiltrud } } Dr. Hiltrud Mueller-Sigmund } Institut fuer Mineralogie, Petrologie und Geochemie } Albertstrasse 23b, 79104 Freiburg (Germany) } Tel.: (+49)-203-6388/-6396 Fax: -6407 } } } hiltrud-at-ruf.uni-freiburg.de (Hiltrud Mueller-Sigmund)
Dr. Hiltrud Mueller-Sigmund Institut fuer Mineralogie, Petrologie und Geochemie Albertstrasse 23b, 79104 Freiburg (Germany) Tel.: (+49)-203-6388/-6396 Fax: -6407
We are looking for antibodies against CD4 and CD8 that can be used with formalin fixed, paraffin embedded rat tissues - with or without application of antigen retrieval procedures. Has anybody some helpful experience ?
Thanks alot.
Heinz
**************************************************************************** Dr. Heinz Fehrenbach Institute of Pathology University Clinics "Carl Gustav Carus" Technical University of Dresden
What is the maximum tilt angle available onto the JEOL JEM 2010 FEG UHR (Ultra High Resolution)? The technical specs in the brochure give us +/- 25=B0? Is there any special TEM sample holder use to reach this values?
I am planning to do immunogold on brassica pollen grains. I am looking for an adhesive that remain tacky when dried and would hold pollen through out the process of washing, fixing, immuno-treatment, postfixing, dehydrating and critical point drying.
Any suggestions will be appreciated. Thanks.
Ann Fook Yang EM Unit, ECORC Agriculture and Agri-Food Canada, Central Experimental Farm, Ottawa, Ontario, Canada K1A 0C6
I'm trying to adapt a resin recipe for use in a processor. Right now, I'm adding 3%BDMA which gives a nice block but polymerizes too quickly. Will decreasing this to 2% slow this down a bit? Yes, I could spend the next 3 days experimenting but I'd like to get some material processed by the end of the week. Many thanks Grace
I would like to see some more conversation on this cost analysis thread.
I am in the middle of trying to justify hiring another tech to help me get out of this back log of work I'm into. The powers to be want me to determine how much time I spend working on each project and investigator. I am spending an exorberant amount of time doing this as I typically do multitasking of different steps on different samples for different investigators essentially at the same time. This they say will tell them what is taking up my time and what another tech would need to be hired for (I already know this info) . Even if I generate the numbers for them they have nothing to compare my workload with to say yes you have more samples coming in than you can take care of. As we know EM is very time consuming. I would like to see what volume of work other department run (but service for entire campus) facilities are producing vs the number of techs producing that work. And because some places may have advanced equipment, it should be limited to producing standard blocks and negative staining techniques. Would any facilities like to help me out on this?
My technique for the cost analysis was to break up the service costs into major steps: processing and embedding, survey sections, thin sections, and scope time. Each step was divided into cost of: materials, and labor, per sample or block where I timed my self performing each step on average. There was also a category for specialized techniques and training of students or others.
Maybe this type of material could be a tutorial or subject at a future MSA meeting ? I was unable to go to the last one that had a round table on facility management (Cincinnati ?). Was this information discussed ?
People looking for costs on equipment should look up the Technologist Forum's Facilities and Equipment list that is available. Thanks for any advice out there.
Rick Vaughn
Electron Microscopy Research Facility Dept Cell Biology & Anatomy Univ Neb Med Ctr RLVAUGHN-at-MAIL.UNMC.EDU
Does anyone still sell the Zero-Stat gun? If anyone has any information on where I can purchase one or if it's still on the market please let me know. Thank you for your help.
Laura L. Estok M.E. Taylor Engineering, Inc. 21604 Gentry Lane Brookeville, MD 20833 Phone: 301-774-6246 * FAX: 301-774-6711 * e-mail: Metengr-at-aol.com
That still doesn't answer the question as to why the other web sites take me directly to the EVEX page. It may not be an intentional deception since I am aware that you service other manufacturers' equipment. But it sure pushes the envelope of what is proper.
At 10:25 AM 11/17/97 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
This looks like a job for activity based costing (ABC) analysis. I suggest that you see if any of your school's accounting students (or professors) need a project. ABC accounting can be fairly complicated for a large organization, but your situation seems manageable enough for a small project team. A broad outline is as follows: list the activities each person does (including non-value-added time such as waiting for results!) with the percentage of time spent on each activity, then divide each person's fully loaded salary by the time spent on each activity (everything should total 100%), then add the costs for each activity. At this point you will have a pretty good estimate of what it costs for each activity. Then map (flowchart) the process including all decision points to show where the money goes and why. From this point, standard TQM tools (storyboarding, Pareto analysis, etc.) can be used to improve the process. It's a lot of work, but if you need to argue a case for more money, the powers that be need something they can see, with dollar signs attached. As far as backlog cost calculations, try to find out from the customer what is not being done (i.e. projects not completed, papers not completed, etc.) and their best estimate on how much their time is worth.
I think if you approach the powers that be on the premise that the objective is not to increase costs by hiring a tech, but rather to improve customer service by allowing the process improvements pay for the tech, then you will have a better chance of getting support. If somebody in a position of power can gather support for the cross-functional effort involved in such a project, then their reputation will be enhanced as well. You could conceivably get the tech you know you need, make your leaders look good, get free/low-cost accounting help (without you doing all the work), employ a deserving technician, help an accounting student get valuable process improvement experience, reduce your backlog, and improve your service to customers. Win/win all around.
my two cent.
------------------------------------------------ Opinions or statements expressed herein, rational or otherwise, do not necessarily reflect those of my employer.
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 Phone: (508) 750-1717 E-mail: crossman-at-osi.sylvania.com
Our web sites: www.sylvania.com www.siemens.com --
When I attempted to reach the IXRF website several months ago, I found myself staring in disbelief at the Evex web page. Thinking I must be totally stupid, I retyped the web address "www.ixrf.com" into Netscape, and there it was again, the Evex web page. At first it reflected badly on the IXRF name, to me.
This was confirmed by Kenny Witherspoon at IXRF, who lamented that they essentially lost their site name to Evex, because one can officially license a site name now, and IXRF had not done so. It is my understanding that Evex scarfed "www.ixrf.com" that was already in use. (To reach IXRF' web page you have to use www.ixrfsystems.com, by the way)
Does Evex claim to represent IXRF, sell IXRF products, etc. What is the story?
Just what does "Evex" mean anyway? As in (K)evex?
Anyways, customers find deceptive strategies similar to this to be a real turn-off, and when they are purchasing equipment, they wonder how this reflects on issues related to service, repair, etc. It is poor business practice and doesn't fool anybody in the end.
Dear Gregory, are you quite sure about your question which seems to be really a = centennial question ??? I made my lession now and the passed 2 hours in studying a lot of = ("old") literature (originals starting in the late 50ies) as well as textbooks of = histology, EM etc.,=20 etc....This problem amazed me since I had a personal question about this = when starting my "carrier" as Electron Microscopist in 1981: you are right in = that there are variable formulas in mixing up "Soerensen=B4s" Buffer solution: sodium = and potassium phosphate (basic, acid) as well as sodium (acid) and sodium (basic) on = the other hand (as well, might be, other substances).
Unfortunately I don=B4t have at hand Soerensen=B4s original paper. If = this is a *must* for you, I could try to get the original publications, since they are = written in "German" journals of the turn of the 19th to 20th century and therefore my be = available more likely in Austrian or German libraries (please let me know).
Follows now a Sherlock Holmes story (maybe only a story by his = assistent, whose name unfortunately I don=B4t remember now):
I didn=B4t read, but had a short "insight" to appr. 10 textbooks of = histology, histo- chemistry, etc., incl. PEARSE A.G.E.(Ed) Histochemistry Theoretical and = Applied, Vol. 1: Preparative and Optical Technology, CHURCHILL-LIVINGSTONE, 4th = Ed,=20 1980 (see p. 236: BUFFERS: pH 5.29 to 8.04: Soerensen (1909-12): Na2HPO4 =
0.06M and KH2PO4 0.06M-mixture) since other book and paper sources = quoted PEARSE (1953) as reference. Unfortunately PEARSE did NOT include a = bibliographic reference for that formula, but, in fact, he mentioned = "1909-12"; o.k. next step: Another paper cited: "Soerensen=B4s phosphate buffer, adapted from = LILLIE and=20 FULLMER ( Histopathologic Technic and Practical Histochemistry, 4th Ed., =
McGRAW-HILL Book Company, 1976)": there I found in Chapter 20: "Buffers and Buffer Tables", p. 878 the = following: } } Table 20-12: Soerensen=B4s phosphate"s" *(ref. for foot note): "the = mixtures on this page were made by P. Jones, and read electrometrically on a Beckman = pH-meter.=20 Readings are corrected to 25 degr. C and slightly smoothed. Phosphate = buffer*s*=20 above 8 and below 5.3 are considered unreliable for histologic use, and = readings are omitted. { { The given formulas in the table are:
Left side: "Dry salt mixtures for field use* (ref. to footnote:): = follows amounts in=20 milligrams of Na2HPO4 + *NaH2PO4.H2O* as well as Na2HPO4 + *KH2PO4*=20 (different milligrams for different pH-values ranging from 5.3 to 8.0). = The footnote pays attention to "The dry salt mixtures calculated from Soerensen=B4s 0.067 = M data. They=20 are to be dissolved in rain water at 1% concentration, ca. 0.070M; if = higher dilutions are used, pH values may be approximated by the following table":
follows grams and milligrams per liter for 0.1 M, 0.067 M and 5 mM, = respectively.
Right side of Table 20-12: } } *Phosphates* at 0.1M, 0.067 M and 5 mM { { The table consists of 5 rows of data: first row (1): second row (2): pH values at specified dilutions (row = 3-5):
In that Chapter 20 a lot of Buffer Formula=B4s one can find. Some = formulas are referenced solitary like: "Gomori(Goemori):personal communication, = ....unpublished" etc. As Main References there are given: W. M. CLARK (Ed): "The determination of Hydrogen Ions", 3rd Ed., = Williams=20 & Wilkins, Baltimore, 1928, and A.G.E.PEARSE (Ed):Histochemistry, 2nd = ed., Little Brown, Boston, 1960. Unfortunately: NO direct reference to an ORIGINAL PAPER of Soerensen.
Therefore: another book:
PERRIN D.D., DEMPSEY B. (Eds): Buffers for pH and Metal Ion Control: = SCIENCE=20 PAPERBACKS # 157 (Chapman & Hall, London, N.Y.), 1974 (1st ed., maybe = there=20 is a newer one now) approx. 180 pp, incl. subject index. Searching the subject index: for } } Soerensen { {: only "Soerensen p*s*H scale" (p. 26) is mentioned: mentioned there (after a statement what the usual standards for = pH-measurement are: (0.05M) K-hydrogen phalate- and (0.01M) Na-borate-buffers and what = "strictly=20 the standards of the US NBS are based on (molalities/moles per kg = solvent) and the=20 British standards are based on (molarity/moles per litre of solution)" = and some=20 information on the concentration vs. buffer capacity there follows a = next paragraph } } Buffer values on "the original Soerensen "p*s*H" scale" can be = converted=20 approximately to the agreed standard scale *by adding 0.04* { {
No word more about *Soerensen* or *Soerensen psH-scale": I checked the = text of the whole book on another quoting of a hint on Soerensen: = nothing.
Next: searching for } } phosphate buffers" { {:=20 references to p. 27: "The pH 7.4 phosphate buffer given in Table 3.3 = (p.41) is useful=20 as a reference in measuring the pH of blood..."...... nothing about=20
*Soerensen*....because Table 3.3. p 41 is entitled: NBS phosphate = buffers as pH=20 standards* (*footnote: BATES 1962): it consists of given mixtures "Soln. = I" and "Soln II", whereby "Soln I" is given as } } 0.025 M KH2PO4 (3.388g) and 0.025M = Na2HPO4 (3.533g) in 1 l of solution at 25 degr.C { { and "Soln II" is given as } } = 0.00869M=20 KH2PO4 (1.179g) and 0.0304 M Na2HPO4 (4.302 g), in 1 l of solution at 25 =
degr.C { {. These solutions at different degrees C make different ranging = pH-values =20 for 0-95 degr. C for Soln I, and 0- 50 degr. C for Soln II, resp.=20 Nothing more about *Soerensen*.
Without notice in the subject index: Table 3.15 (p.53, but indexed by = "phosphate=20 buffers"): } } pH values of isotonic Soerensen*(footnote) buffers+(footnote) { {: =20
The formula uses "Na2HPO4 and NaH2PO4.H2O or NaH2PO4.2H2O, respectively, the final mixture also containing NaCl (mg/100 ml) to adjust the = tonicity to 0.92 at 37 degr.C". pH values are given for 25 degr.C as = well as for 37 degr.C and differ at same=20 pH levels/mixtures in the range 0.01 - 0.02 (e.g. pH -at- 25 degr.C =3D = 5.76, -at- 37 degr. C =3D 5.74; -at- 80 degr.C. 7.33, -at- 37 degr. C 7.32) footnote *: refers to SOERENSEN (1909), footnote "+": mentions CUTIE &=20 SCIARRONE (1969): No hint on "KH2PO4 and/or Na2HPO4......."
Next step: searching the "references index" for "Soerensen": uncredible, = there it is: !!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!= !!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!= !!!!!!!!!!!
Soerensen (1909): no title, "Biochem. Zeits." (which means "Biochemische =
Zeitschrift"), *21*, p. 131 & !!!!!!!!!!!BUT!!!!!!!!!!!!
there is another reference too:
Soerensen (1912), no title, "Ergebn.Physiol." (which means "Ergebnisse = [in] der=20 Physiologie"),*12*, p. 393 &
Now I thought that there had to be another, second "Soerensen" reference = within the=20 text or tables. Subject index for "phosphate buffers" in addition said: = p. 138, 139,=20 151:
} } For disodium hydrogen citrate, NaOH, HCl buffers covering the pH = range 2.2 -=20 6.8, see *Soerensen (1909, 1912)* { {
Got it.
So at the end, despite not owning the original papers of Soerensen, I = think those=20 must be experimental papers dealing with concentrations, temperatures, = dilutions,=20 diverse buffer substances (according to PERRIN/DEMPSEY 1974, p. 1, the = term=20 "buffering" was created by FERNBACH and HUBERT 1900, cf. Compt. rend. = *131*,=20 p 293 &) and the so called "psH" -scale of Soerensen is most probably a = protocol of=20 his measurements, dealing also with a variety of phosphates (at least K- = and Na- PO4), which sometimes is referred to as "Soerensen=B4s buffer" for = sympathetic, "historical", "respectful" or "ancient" reasons, but still = is 0.1 M or 0.067 M or =20 0.005 M buffer (see above) exhibiting slight modifications in pH and = maybe in buffer=20 capacity by concentrations as well as temperature. Some authors=B4 tables or their infos I went through state there is no = or even little=20 difference in using Na-Phosphates instead of K-Phosphates, provided that = the molar ratios are kept.
So I wouldn=B4t worry about either *Na-Na-PO4* or *KPO4 -NaPO4* = ingredients. Most important conditions for me are buffer capacity at a given = concentration and +/- isotonicity (which sometimes has to be increased = by adding substances like glucose, sucrose or even NaCl). I am using = Millonig=B4s 0.13M and 0.10M for buffering my specimens, osmication = and/or washing solutions. Millonig=B4s buffer solution is either prepared by mixing Na-Na-PO4 or also formulas using only NaH2PO4 = (acidic) and NaOH (basic, alkaline component; Formula, if wanted, on = request).
Just for completion of the reference list I give the reference for = GOMORI 1955,=20 because he was obviousely a very famous scientist and worker in that = field=20 (histochemistry and histochem. staining reactions), maybe that book is = more readily accessable to you (library) and you can find there valuable = informations:
GOMORI (1955): Methods in Enzymology *1*, p. 141 &.
Hope this helps, if I should have a look for getting those (certainly) "German written" = original papers (which will be a little bit difficult, but.....and it = will take some time to get it),=20 please don=B4t hesitate to send me your e-mail request.
Best regards to you and yours,=20 have a nice day
Dr. Wolfgang MUSS Department of Pathology, LKA EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")=09
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
Question/debate of the day:
What is the actual formula Sorensen used to make Sorensen's = phosphate buffer? Not what you think it is, but What Dr. Sorensen stated in = his paper in 1909, and what many of us have or thought we have been referencing for the past 88 years.
Some references say (Hayat) to use sodium phosphate and Potassium phosphate
Most other references state Sorensens Phosphate buffer has being = made with sodium monobasic and sodium dibasic salts.
If anyone has the original reference to sorensens (biochem, 22, = 253, 1909) on hand, I would be most appreciative if they would be kind enough to email or fax it to me.
Thanks in advance Gregory Argentieri Novartis Pharmaceuticals Corp 59 Rt 10 East Hanover, NJ 07936
Dear Greg, unfortunately I found one data jam in my mail to you:
In the paragraph 8 lines above !!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!= !!!! (followed by the original Soerensen references) instead of:
The formula uses "Na2HPO4 and NaH2PO4.H2O or NaH2PO4.2H2O, respectively, the final mixture also containing NaCl (mg/100 ml) to adjust the = tonicity to 0.92 at 37 degr.C". pH values are given for 25 degr.C as = well as for 37 degr.C and differ at same=20 pH levels/mixtures in the range 0.01 - 0.02 (e.g. pH -at- 25 degr.C =3D = 5.76, -at- 37 degr. C =3D 5.74;=20
-at- 80 degr.C. 7.33, -at- 37 degr. C 7.32)
footnote *: refers to SOERENSEN (1909), footnote "+": mentions CUTIE &=20 SCIARRONE (1969): No hint on "KH2PO4 and/or Na2HPO4......."
Next step: searching the "references index" for "Soerensen": uncredible, = there it is: !!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!= !!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!= !!!!!!!!!!!
it should be read:
"-at- 25 degr. C. 7.33, -at- 37 degr. C. 7.32" Sorry! Bye W.
To all concerned: Here at the Bessey Microscopy Facility at Iowa State University, we have determined the costs for all of our operations and what to charge our users for the equipment, tech services, etc. If anyone wishes a copy of our charges and/or basis for the figures, please send a FAX # or address and I will send you the information. Cheers! Bruce Wagner
} November 17, 1997 } } It is not Evex Analytical's intention to deceive anyone. } } Promotion of Evex Analytical Inc., is promoted only at } www.evex.com } } Evex Analytical does own many other websites but promotes its products at } www.evex.com } } http://www.evex.com/971117.htm
So how come you're insufficiently upfront to personally sign your emails to the listserver?
While I'm on the subject, there have been a few postings recently signed only with a nom-de-plume. Is this ok? Doesn't seem quite reasonable to me.
cheers
Ritchie
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
Oops, Evex was just deleted from our site of microscopy related links. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au } } } Well, I really don't usually follow up on *myself*, but I went ahead and } did a WhoIs on EVEX. It turns out that EVEX, happens to have registered } the following names: } } EVEXTG.COM } TNSERVICE.COM } MEKTECH.COM } IXRF.COM } EDAX-EDS.COM } NORAN-EDS.COM } THOMSON-SCIENTIFIC.COM } EVEX.COM } } } billo
There is a product called Mikrostik sold by Ted Pella, Inc. (800-243-7765 is the toll-free number for Canada). Other vendors might sell this, also. This is designed for small particle adhesion on stubs and is not supposed to bury the things, like some tapes and tabs do. I have used it successfully a couple times for things like phytoliths. I have no idea if it would survive the washing, fixation, etc. processes, but it might be worth a try.
I have no financial interest, etc.
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
} -----Original Message----- } From: METENGR-at-aol.com [SMTP:METENGR-at-aol.com] } Sent: Monday, November 17, 1997 9:57 AM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: Zero-Stat } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } ---------------------------------------------------------------------- } -. } } Hello all--- } } Does anyone still sell the Zero-Stat gun? If anyone has any } information on } where I can purchase one or if it's still on the market please let me } know. } Thank you for your help. } } Laura L. Estok } M.E. Taylor Engineering, Inc. } 21604 Gentry Lane } Brookeville, MD 20833 } Phone: 301-774-6246 * FAX: 301-774-6711 * e-mail: Metengr-at-aol.com
I am looking for a supplier who carries Hessian Bordeaux. This stain is reportedly used to differentiate damaged starch. Any information regarding the staining protocol would also be appreciated.
Thank you,
Diana Kittleson Pillsbury Technology East dkittleson-at-pillsbury.com
} } It is not Evex Analytical's intention to deceive anyone. } } } } Promotion of Evex Analytical Inc., is promoted only at } } www.evex.com }
Well then why when these other "websites" are visited, which just happen to contain your competitor's brand names, do we end up AT YOUR HOME PAGE which is promoting "your" products? That's not deceptive I suppose.
} } Evex Analytical does own many other websites but promotes its products at } } www.evex.com }
Put "own" in inverted commas. The web page you get using your competitor's names in a URL is identical, at least to the limit of my ability to distinguish between two images on a PC screen, to the web page you get using www.evex.com. The links are the same too. So how could I be reasonably expected to know that your intention is only to promote your products through www.evex.com.
Your Press Release is an insult to our collective intelligence.
} November 17, 1997 } } It is not Evex Analytical's intention to deceive anyone. } } Promotion of Evex Analytical Inc., is promoted only at } www.evex.com } } Evex Analytical does own many other websites but promotes its products at } www.evex.com } } http://www.evex.com/971117.htm
There is more....
In the context of the preceeding messages in this thread, the curious might want to take a look at screenshots of the Evex "VIDX X-ray Microanalysis System" that can be viewed at either of the following identical websites, both "owned" by Evex:
http://www.evex.com/anal.htm
and
http://www.thomson-scientific.com/anal.htm
and the screenshot of the WinEDS Version 3.0 microanalysis system marketed by the real (and I imagine the original) Thomson Scientific at the following totally unrelated website:
http://werple.net.au/~tsi/brochure.htm
Not making any allegations here, but WHAT GIVES?? Spooky stuff.
Dear Microscopists: I have not noted any replies to the question posed and expect this is because Spurrs' has four components and two (ERL and DMAE) are regarded as fairly toxic. Information about these chemicals is given in our online MSDS. The bigger problem is that some people apparently become allergic and the symptoms of allergies can vary a great deal and the allergy could be in response to different components as well.
I have no idea how common allergies to Spurr's components are, but I have never personally met anybody who had such an allergy. Allergies can be very troublesome but they do not reflect on the toxicity of a product. An (off-topic) example: Our mangos are about to ripen which reminds me that among "mango eaters" about 1 in 50 people develop a progressively nasty allergy. Ripe mangos are excellent food, but to some people they are disastrous.
Toxicity (and carcinogenicity, ERL) of Spurr's is undisputed and basic lab technique should include use of a fumehood and wearing gloves. Clean techniques should avoid wetting the gloves with the liquid resin. Using good working practices should reduce any absorption to such a tiny dosage that non-allergic people should worry about diesel fumes, sunlight, excess iron, lack of certain vitamins and, if life-expectancy was the issue, a drastic reduction of food intake.
While other embedding resins may be a little less toxic, it should be appreciated that all epoxies (including household glues) are toxic. The point is that you as lab personnel have the facilities and the knowledge to dramatically reduce if not eliminate any dangers; spare a thought for the multitudes not in that position and weigh dangers in the lab against activities in your spare time.
I do not criticise microscopist for trying to learn about lab risks; quite the opposite. However, I urge to 1. take precautions and 2. see it all in perspective. Cheers? Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au
} } } I received this inquiry today, and wondered if any of you could help. } } Please respond directly to Margaret, and not to the listserv. } } } } } } At the suggestion of my doctor, I have been searching the internet } } } for information on the long-term effects of exposure to Spurr Resin. } } I'd like to see the responses to this query on the listserver, in addition } to the direct response, please. } } } Caroline Schooley } Educational Outreach Coordinator } Microscopy Society of America } Box 117, 45301 Caspar Point Road } Caspar, CA 95420 } Phone/FAX (707)964-9460 } Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html } Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
Crossman, Harold wrote (in part): } Rick, } } This looks like a job for activity based costing (ABC) analysis... but } your situation seems manageable enough for a small project team..... } From this point, standard TQM tools (storyboarding, Pareto analysis, etc.)....
Yeah, and Rick, don't forget to have your "small project team" of consultants factor ABC and TQM into the cost analysis. Then, PDQ, you'll be right up there with the "powers that be" in management intellegence. Maybe they'll even give you your own key to the Executive WC!
(I'm sorry. This was just too good for someone with a slightly twisted sense of humor to pass up! All the best, sincerely, to everyone. Even you, Harold. ;-) ) --
Stephen A. Shaffer MicroDataware sshaffer-at-microdataware.com (business) http://www.microdataware.com (temporarily out of service) steve_shaffer-at-compuserve.com (personal) http://ourworld.compuserve.com/homepages/steve_shaffer/
I agree with Jim Darley that toxicity and carcinogenic properties are well-specified for Spurr's and other chemicals used in TEM, but that allergies are less easy to define. In my own case, I have become allergic, or perhaps I should say, sensitive (over about 15 years) to aldehyde fixatives, most resins - including Spurr's - and their components and also to darkroom chemicals, the latter so much so that I can do only very limited darkroom processing at one stretch. The visible effects are like sunburn; red, tender, itchy skin on face and hands or any part not well-covered by clothes (or gloves). Takes a couple of days to fade. This is despite taking precautions - only work in fume hood, always wear gloves, etc. It's a good thing we do mostly freeze-sub work these days...
And some of us (e.g. yours truly) are indeed sensitive to a seemingly ridiculous range of things besides the TEM chemicals - paint, petrol and other oil-based organic solvents and creams, car fumes (yep), most perfumes, etc. etc.
I'd guess that allergic symptoms are quite variable, and that people have quite a range of susceptibilities. Haven't looked but also suspect there may not be much literature on this - other than anecdotes like mine. Vague memories of a similar discussion about maximum safe limits of exposure to various toxic stuff, and about people developing allergies, on this list not long ago. You can never take too many precautions....
cheers,
Rosemary White Department of Biological Sciences Monash University, Melbourne, Victoria 3168, Australia phone 61-3-9905 5670 fax 61-3-9905 5613 email r.g.white-at-sci.monash.edu.au
The maximum tilt angle available on the JEOL JEM-2010 F URP is + - 25=B0 = even on the edge of the grid. This holder is quite new, but it have the specifications for the = standard double tilt holder. The part number of this holder is EM-31031.
The system used to fix the sample on the holder is also different : it = is now easier .
R=E9gis RAVELLE-CHAPUIS Application Engineer
ravelle-at-jeol-sa.worldnet.fr
JEOL (Europe) S.A. All=E9e de Giverny Espace Claude Monet 78290 Croissy sur Seine FRANCE
The maximum tilt angle available on the JEOL JEM-2010 F URP is + - 25=B0 = even on the edge of the grid. This holder is quite new, but it have the specifications for the standard double tilt holder. The part number of this holder is EM-31031.
The system used to fix the sample on the holder is also different : it is now easier .
R=E9gis RAVELLE-CHAPUIS Application Engineer
ravelle-at-jeol-sa.worldnet.fr
JEOL (Europe) S.A. All=E9e de Giverny Espace Claude Monet 78290 Croissy sur Seine FRANCE
I have calculated the costs for our different equipments and they can be found on our web-page at: URL http://www.utu.fi/med/em/fees.html The currency is FIM and approximate exchange course is 1USD=3D5,30FIM There are two categories for prices: A subvented price for university research and another for "outsiders" (this includes also wages). The costs are calculated for 5 years lifetimes of the instruments. Service costs, energy, room rents, etc. are included.
I hope this will help
Regards, Jouko
Jouko K. M=E4ki, Ph.D., Laboratory Manager University of Turku, Laboratory of Electron Microscopy Kiinamyllynkatu 10 FIN-20520 TURKU FINLAND Tel: + 358 (0)2 333 7318 GSM: + 358 (0)40 505 2521 FAX: + 358 (0)2 333 7380
I have calculated the costs for our different equipments and they can be found on our web-page at: URL http://www.utu.fi/med/em/fees.html The currency is FIM and approximate exchange course is 1USD=3D5,30FIM There are two categories for prices: A subvented price for university research and another for "outsiders" (this includes also wages). The costs are calculated for 5 years lifetimes of the instruments. Service costs, energy, room rents, etc. are included.
I hope this will help
Regards, Jouko
Jouko K. M=E4ki, Ph.D., Laboratory Manager University of Turku, Laboratory of Electron Microscopy Kiinamyllynkatu 10 FIN-20520 TURKU FINLAND Tel: + 358 (0)2 333 7318 GSM: + 358 (0)40 505 2521 FAX: + 358 (0)2 333 7380
Electron Microscopy Technician - EM technician needed by state-of-the-art laboratory studying the organization of proteins and nucleic acids in cell nuclei. The individual should be competent in thin sectioning with a diamond knife, fixation and embedding methods, darkroom work and have some experience in imunofluorescence microscopy. Experience in cell culture is also desirable. Please send resume and two letters of reference to: Dr. David L. Spector, Cold Spring Harbor Laboratory, P.O. Box 100, Cold Spring Harbor, New York 11724.
***********Please do not respond to this e-mail address.******
Fellow Netters, Someone has "borrowed" my Frieda Carson book and Sheehan does not have the procedure on how to prepare Zinc Formalin. I know most of the labs that I have spoken to locally and have switched to it, purchase it commercially (Anatech). I am getting ready to give a Histotechnology Workshop/Course in Panama. And in previous Workshops in Central and South America one question has begun to be asked, "How does one prepare Zinc Formalin" I hope to have the answer incase I am asked again this time. Thank you for all your help, Teresa
I have been watching the thread on website parasites and website parking with great interest, as BUEHLER has also gone through a similar situation. Since this does relate to your ability to access our website and gather information on metallographic and microscopy products, I think it is appropriate to mention here.
Two years ago, when BUEHLER tried to register our trademark name, BUEHLER.COM, we discovered that the name had been purchased by a company called MOZES, CLEVELAND & CO. This is a website hosting firm. They claimed that they had purchased the name for a company called Buehler Books (which does not exist, and the domain name has never been actively in use). This seemed innocent enough, until we discovered that MOZES, CLEVELAND & CO. had registered the name LECO.COM; another metallographic supplier (LECO has subsequently purchased their name back from MOZES, CLEVELAND & CO.). This all seemed highly coincidental, given the fact that the supposed company, Buehler Books was not a metallography supplier.
We then discoved that our main competitor, STRUERS, was actively doing business with MOZES, CLEVELAND & CO., using their services to host the STRUERS website. More coincidence! Turns out that the name STUERS.COM is owned by STRUERS, but is administered, wholly, by, you guessed it, MOZES, CLEVELAND & CO.
I do not in any way suggest that STRUERS had something to do with MOZES, CLEVELAND & CO. purchasing the domain names (trademarks, I might add...) of their two chief competitors. This could have been a clever idea of an independent company who did their research on one of their client's competitors. However, this does not change the fact that our trademark is unavailable to us for use as a domain name.
We have addressed the legal situation with MOZES, CLEVELAND & CO., and have received a decidedly chilly response from the owner, Jeno E. Mozes (webmaster-at-clevelandoh.com). He has offered to sell our name back to us for an outrageous fee, which seems to suggest that his story about Buehler Books was a bit of un-truth. We have not ruled out further legal action in this matter.
If you have searched for BUEHLER by typing www.buehler.com, you have been looking in the wrong place. You can find us at: www.buehlerltd.com
Sincerely, Scott D. Holt BUEHLER, LTD. PO Box 1 41 Waukegan Rd. Lake Bluff, IL 60044 (847)295-6500 http://www.buehlerltd.com
If metal nanoparticles (~1 nm) would be appropriate for tagging the substrates, we can label some of the substrates with our 1.4 nm Nanogold cluster label, which we have cross-linked to a number of small molecules before (hope this doesn't sound too much like a commercial plug - we are interested in this type of experiment from a research perspective as well).
Rick Powell Nanoprobes, Incorporated
****************************************************************** * PLEASE NOTE MY NEW E-MAIL ADDRESS: rpowell-at-mail.lihti.org * * NANOPROBES GENERAL E-MAIL ADDRESS: nano-at-mail.lihti.org * * * * NANOPROBES, Incorporated | Tel: (516) 444-8815 * * 25 East Loop Road, Suite 124 | Fax: (516) 444-8816 * * Stony Brook, NY 11790-3350, USA | nano-at-mail.lihti.org * * * * NOW EASY TO FIND ON THE WEB: http://www.nanoprobes.com * ******************************************************************
Dear "freeze substitution microscopists", is ther any facility, where we could do some (not many) high pressure freezings of plant roots and/or a fungus? We could do the further preparation in our labs, but it makes not much sense to freeze by "conventional" methods, to much ice crystals. Anybody interested in such a kind of cooperation in Germany (or countries near to Germany)??? Arthur Dr. Arthur Schuessler University of Heidelberg Zellenlehre D-69120 Heidelberg Germany
HI everybody Althought this message has nothing to see with Microscopy It's very interresting for an European (we are a bit late on Internet) to see the fight for the names of Web sites. Shall we rename World War Web? I'm not surprise that some genius has licensed some name in order to hack or to sale in a next future to the involving company. Just have a look on what you think is the web site of KLM (Dutch airline company) http://www.klm.com and you will be surprise of the link. I guess this name was licensed and KLM buy the right to link to his official site was bought http://www.klm.nl Anyway that kind of practices are not very commercial /or for money maker but who can complain in that word of monkey businees we've created. Of course this is my opinion and not my company opinion....etc Regards ========================================================== Jacky Larnould mailto:larnould-at-worlnet.fr voice:33 (0)4 67 72 28 26 fax :33 (0)4 67 79 54 90
Friends, A peer is looking for a inert heat conducive material for layering onto silica wafers with sub-micron bumps to make this bumpy surface smooth. The reason is to accurately measure temp with a smooth surface probe (smooth to smooth surface for accurate measuring). The conditions are 300 celsius and under vaccuum. Here is the problem: once the measurement is done, the material must be removable leaving absolutely no trace, exposing once again the bumpy surface. Are we dreaming or is there such a material out there? All responses (fictional or non) will be appreciated.
I wanted to take the time to thank everyone who contacted me regarding= my Reichert OmU2 woes. At this time I have passed along the suggestions= to my crack repair team who are on it.
In the meantime, while we were rummaging through the lab in search= of the elusive yet non existent spare belt, my archaeology team= discovered the eighth wonder of the Chemistry World. This would= be in the form of a silvered glass vacuum column, about a meter= long, with an outside taper on the bottom, inside taper on the top,= and through intentionally missing patches of silver on the side= one can see some type of distilation array running down the length= of the middle. "Oldershaw column" is written on the box and the= only other markings are HGF from Stafford TX on the tube. I looked= in American Laboratory but HGF isn't there, so since it's probably= older than I am, can anyone tell me what this is? It looks new but= for all I know was used on the Manhattan Project, or at least during= that epoch.
Thanks again,
Laura
************************************************************ Everyone has a photographic memory. Some don't have film. ************************************************************ Laura Rhoads Electron Microscopy Facility Director Department of Biology Western Kentucky University 1 Big Red Way Bowling Green, KY 42101-3576
The following WEB sites all have the same IP address of 165.254.117.137 www.evex.com www.ixrf.com www.mektech.com www.noran-eds.com www.thomson-scientific.com
There are several poyimides and photoresits available that are used for "planarizing." They all can be easily removed, but the amount of planarization depends on the size of the bumps. 300 degrees C should not be a problem.
best regards mark
Mark W. Lund, PhD Director } } Soft X-ray Web page http://www.moxtek.com { { MOXTEK, Inc. 452 West 1260 North Orem UT 84057 801-225-0930 FAX 801-221-1121 lundm-at-xray.byu.edu
"The state is good at simple tasks, like killing people and seizing their wealth. It has far more trouble reaching inside individuals and making them good." Doug Bandow
In the early '90's Balzers had a subsidiary called Bal-Tec which sold EM accessories. In particular a flat (45 mm) round (190 mm diam) desiccator for storing SEM stubs cat No. BU 014 053-T. Now I want some more, I find Balzers no longer sell them.
Has the BAL-Tec agency passed into other hands? Does anyone else sell these? It will help our storage situation greatly.
TIA,
Mel Dickson Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
Approximately 2 years ago my institution received a Philips EM 300 as a donation. After 2 years of cutting through miles of red tape I am finally ready for installation. Upon uncrating and opening all of the boxes, we discovered that the film loading and receiver boxes are both missing along with the photographic plate carriers/holders (31/4 x 41/4"). In addition, a tool kit was missing - I was informed that the most important component is a long rod-like tool with a square endpiece for working on the vacuum system.
Does anyone know whether these items are still available (I intend to inquire at Philips) or would anyone be willing to donate or sell these items from an old Philips which has been put out of service. Am I limited to EM 300 boxes and plate carriers or are they interchangeable with other Philips TEM's.
Any assistance/suggestions would be greatly appreciated!
Stephen J. Beck Associate Professor Bio-Imaging Center/Electron Microscopy Department of Biology Nassau Community College Garden City, NY 11530 Voice Mail: (516) 572-7829 Email: {becks-at-sunynassau.edu} URL: {http://www.sunynassau.edu/webpages/biology/becks.htm}
Hello Mel, They do exist in Lichtenstein. Technotrade represents them in the USA. You may contact Mr. Auwaerter for the details by email: AAuwaerter-at-aol.com. Sincerely, Marek. PS Thanks for the reprint, just arrived.
representation} ---------------------------------------------------------------- -------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello Mel, They do exist in Lichtenstein. Technotrade represents them in the USA. You may contact Mr. Auwaerter for the details by email: AAuwaerter-at-aol.com. Sincerely, Marek. PS Thanks for the reprint, just arrived.
representation} ---------------------------------------------------------------- -------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Teresa, maybe meanwhile you got the formula for mixing up Zinc Formalin. If not: here you get one:
1% Zinc Sulfate in 10% unbuffered Formalin, pH 3.7
Ref.:=20 HERMAN, CHILIPALA, BOCHENSKI, SABIN, ELFONT: Zinc Formalin Fixative for = Automated Tissue Processing: J. Histotechnol. Vol. 2/No.2, June 1988, p. = 85-89
BANKS, CARON: The use of Metallic Salts as an Adjunct to Formalin For = Routine Tissue Fixation; Mayo Clinic, Mayo Foundation (unpubl. summary = of Results)
If you want to have a Protocol for a hand-out on: Restoration of Estrogen Receptor Sensitivity in Formalin-fixed, = Paraffin-Embedded Breast Carcinomas, Procedure for Restoring Antigenicity of Over-Processed Formalin-Fixed = Paraffin Blocked Tissues (Antigen Retrieval) then please send your Fax number via E-mail .
Hope this helps, best regards
Dr. Wolfgang MUSS Department of Pathology, LKA EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
---------- Von: Flores, Teresa[SMTP:tflore-at-lsumc.edu] Gesendet: Dienstag, 18. November 1997 16:19 An: histonet-at-pathology.swmed.edu Cc: microscopy-at-sparc5.microscopy.com Betreff: Q:Fix: Zinc Formalin, formula
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
Fellow Netters, Someone has "borrowed" my Frieda Carson book and Sheehan does not have the procedure on how to prepare Zinc Formalin. I know most of the labs that I have spoken to locally and have switched = to it, purchase it commercially (Anatech). I am getting ready to give a Histotechnology Workshop/Course in Panama. And in previous Workshops in Central and South America one question has begun to be asked, "How does = one prepare Zinc Formalin" I hope to have the answer incase I am asked again this time. Thank you for all your help, Teresa
Due to consolidation processes you maybe will be informed by the company = listed below the name of "Balzers" and "BAL-Tec" (for Vacuum sector) has = changed to:
PFEIFFER Vacuum Ltd. Their representatives in Austria/Europe sell, besides the "BALZERS = Instruments"-Components, in Austria "exclusively" also the "complete = program" components sold hitherto under " BALZERS OPHTHALMIK AG", = specimen preparation app. for EM of the "BAL-TEC AG" as well as the = "vacuum-metallurgic apps." of the "PFEIFFER VAKUUMANLAGENBAU GmbH".
I own a PFEIFFER "VACUUM TECHNOLOGY 1997"-Cat (German ed.) which shows = the USA-Representative
PFEIFFER Vacuum Technology Inc. 24 Trafalgar Square NASHUA NH 03051 USA
phone: +603+578-6500 fax: +603 578-6550 unfortunately, no e-mail is given
I think, they should sell the old programme of BALT-TEC, you need.
Good luck, best regards
Dr. Wolfgang MUSS Department of Pathology, LKA EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
Hello world,
In the early '90's Balzers had a subsidiary called Bal-Tec which sold EM accessories. In particular a flat (45 mm) round (190 mm diam) = desiccator for storing SEM stubs cat No. BU 014 053-T. Now I want some more, I = find Balzers no longer sell them.
Has the BAL-Tec agency passed into other hands? Does anyone else sell these? It will help our storage situation greatly.
TIA,
Mel Dickson Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
Does anybody know of any free or shareware software for processing microscopy images that allows, for instance, to distinguish between grains in metallic materials or determine the phase contents of the material, etc.?
The image source is the digital camera or scanned photography image.
I would very much appreciate having any information in this or related fields.
I am afraid that you are just partly right at the Bal-Tec story. Bal-Tec was bankrupt more than a year ago. Then a new owner has bought the properties of the former firm during the consolidation process. Then ha says, according to the law in Lichenstein he was able to launch a new firm under the same name as earlier: Bal-Tec. So, there is a Bal-Tec firm which produces EM accecories and units and this is different form Balzers-Pfeiffer which is based in Austria. Their addresses:=20 BAL-TEC AG F=F6hrenweg 16 Postfach 62 FL-9496 Balzers F=FCrstentum Lichenstein Tel: 41-75-388 12 12 Fax: 41-75-388 12 60
in US: TECHNOTRADE 7 Perimeter Road Manchester NH 03103-3343 Tel: 1-603-622 5011 Fax: 1-603-622 5211
Sincerely Yours, Bela Pecz 19th November 1997 ----------------------------------------- Dr. Bela Pecz Research Institute for Technical Physics H-1325 Budapest, POBox 76 Hungary phone: 36-1-2332 865 fax: 36-1-2332-794 E-Mail: pecz-at-mufi.hu -----------------------------------------
I've been asked to distinguish volcanic ash from Saharan desert dust in particles collected on air filters using SEM/EDX. I have requested samples from both sources to provide "source signatures", but don't know if these will be available. In the meantime, are there morphological or chemical differences which can distinguish these two types of particles? Thanks very much for your suggestions!
Thanks to those who expressed their concern about the website parking issue. I mentioned it because it seems to be a rather common practice, and it interferes with the free flow and sharing of information.
Although a few people have sent me suggestions as to how I might 'adjust' the situation, and I thank them, I think spamming the offenders email may not be anything more than sinking to their level. And, it might backfire! As Nestor pointed out, spamming from listserver users might cause a response which would overload the server.
I wanted to inform...not to incite. We will continue to persue legal means of regaining use of our trademark.
Thanks to all.
Sincerely,
Scott D. Holt BUEHLER, LTD. PO Box 1 41 Waukegan Rd. Lake Bluff, IL 60044 (847)295-6500 http://www.buehlerltd.com
We are doing whole-mount immunofluorescence of walled fungal cells, and have run into problems of permeability of a IgM antibody we wish to label with. We have no problems with our IgG antibodies, but this IgM simply does not get in (at least not consistently and evenly). I was told that it may be possible to digest the IgM with trypsin to generate Fab fragments. I was also told that it is difficult to control the activity of the protease to prevent unwanted cleaving. Does anyone have any experience with this approach? A reference with a protocol would be greatly appreciated!! Likewise, any additional advice/comments would be welcome. Thanks.
Tim Bourett DuPont Experimental Station Wilmington, DE USA
Hi, dear colleagues! For radial distribution function (RDF) method I need a receipt to take coherent part of electron scattering curve. Any suggestions or literature references (journals, conferences, etc.-it is better) would be appreciated. Thank you very much for attention to my problem.
} } } Although a few people have sent me suggestions as to } how I might 'adjust' the situation, and I thank them, I think } spamming the offenders email may not be anything more } than sinking to their level. And, it might backfire! As Nestor } pointed out, spamming from listserver users might cause a } response which would overload the server.... }
I would like to pipe up with some agreement here. Using bad net behavior in response to bad net behavior is a lose-lose proposition. In the non-commercial arena, it is pretty much useless and indefensible. In the commercial arena, it is useless and often actionable.
For the most part, the best response is simple openness. When people act like asses, the ethical status of the behavior is pretty obvious *and* they don't want it known. Since the actions speak for themselves, it is not necessary to run around and say "This is bad!!" It is only necessary to point out that "This is."
In the case of domain-name parasites, one really only needs to make sure that *everyone* knows what the parasite is doing. The very act of parasitism then becomes its own anti-advertisement -- or if not, it becomes its own defense. In the case at hand, for instance, it is not really necessary for a person who thinks that EVEX's use of www.mektech.com is real parasitism to say so, and EVEX's denial is equally unnecessary. The act speaks for itself. If one really cared about it, one need only bruit it about without comment, perhaps in a sig line.
If it is obviously parasitic, the result will be negative. People on the net react so strongly to parasitism that the downside will overcome any positive effects of misdirection. If it is not obviously parasitic, then there will be appropriately little reaction. For instance, let's say I posted with the sig:
"Beware of domain-name hoarders! The following registrations use my name:
oliver.com belongs to George Oliver billo.com belongs to Bill O'Brien oliver.net belongs to "Mailing Services, Inc."
Which one is hoarding names, do you think?"
I don't think that George Oliver or Bill O'Brian really need to run out to defend themselves here (the names above are approximations, by the way).
If, on the other hand, one starts playing nasty games, these things can escalate. For an example of how bad it can get, take a look at
} ... } I've been asked to distinguish volcanic ash from Saharan desert dust ...
The volcanic ash should primarily be amorphous glass, whereas the dust should be the result of physical weathering and abrasion and therefore be relatively resistant minerals. SEM/EDX should therefore show similar "glassy" composition for all particles whereas the dust would show a variety of (eg) qtz, feldspar and oxides. I would have to believe XRD would be the best analytical method and much quicker.
... hope this helps ...
cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
} ... } } Does anybody know of any free or shareware software for processing } microscopy images ...
You don't mention your computer preference but if you follow some of the links provided at: {http://www.ou.edu/research/electron/www-vl/} I think you'll find what you're looking for ...
cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
Thanks to all who responded to my recent question. The responses I received came from what appears to be a good cross section of the EM population, covering university, industry and government users as well as EM suppliers.
Some folks said that there were a number of 20-30 year old scopes which were still in active use, were in good shape and working well.
Most defined technical obsolescence (i.e., the newer equipment available had significantly improved capabilities) as 7-10 years, while operational obsolescence (frequently required, difficult to get parts and service) was defined as 12-15 years.
Most folks who were lucky enough to be able to replace their EM's regularly used a 10 year depreciation schedule for accounting purposes. One person suggested a 5 year schedule for SEMs and a 10 year schedule for TEMs to address the fact that there have been more advances in a shorter time frame in the development of SEMs than TEMs.
In an interesting twist, a couple of people talked about the psychological phenomenon where users or collaborators are more keen to do their work, or have their work done on "new" equipment rather than "old" equipment. I was particularly interested in this phenomenon, since we are being urged by our Administrators to attract Industry collaborators, and this may prove to be an important point in this regard.
I have saved the responses I have received, and will submit them along with my request for new EM equipment. Thanks for the ammunition and the informative discussion.
Paula.
Paula Allan-Wojtas Food Microstructure Specialist Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
There is enough literature out there on "dust" and "volcanic ash" to choke a horse, so to speak.....I guess for you it's a matter of where to start. Here are some tips:
1. the morphology of volcanic ash and "dust", the latter of which could comprise - in part - volcanic ash, are very different, for example:
a. volcanic ash largely derives from explosive fragmentation of magma (better known as lava when emitted to earth's surface as a lava flow) - accordingly textures reflect this process......seeing as the mechanical energy for this explosion derives from rapid decompression of magma and attendant gas exsolution, you should expect - almost always - to find remnant outlines of gas bubbles (vesicles).....also, because the magma is quenched by the process, the ash is glassy (+/- crystals)...so, basically you've got broken glass with bubbles and/or bubble outlines (also, glass often breaks with conchoidal fracture)
b. "dust" on the other hand, if it does not contain volcanic particles, should comprise mainly minerals and/or mineral fragments derived from the surface weathering of rocks.....the overwhelming remnant component of weathering is quartz (silica = SiO2), but what you see depends on both the weathering environment and available source rock for the "dust".....I suppose if one had to generalize, it could be said that "dust" should comprise mainly mineral fragments with surface mechanical defects derived from a combination of chemical etching (chemical weathering, as in dissolution) and mechanical collisions (as generated during movement by wind and water = often recognizable as mini-impact features)....sometimes chemical alteration products of primary minerals are retained on mineral surfaces (as, for example, Fe-oxide or Fe oxyhydroxide might be.....or manganese oxide = desert varnish), so things get tricky
2. as far as chemistry goes, generalizations can again be made:
a. volcanic ash, as indicated above, comprises glass and crystals......the latter could be very similar to minerals derived from (or liberated by) the weathering of rocks, whereas glass on the other hand is fairly distinct compositionally from minerals (save for a few "trash" minerals, like amphibole group minerals, that are "kitchen sinks" that contain up to 10 elements in fairly important abundances)....look for the following
1.magma that produces ash by explosion is somewhere from:
"andesitic" in composition, meaning that it probably has } 55% SiO2, with important levels of Al2O3 (approx. 15%, or more), FeO and MgO (10%-15%, between them), CaO (5%-10%), TiO2 (2%) and Na2O and K2O (4%-6%, between them)
to
"rhyolitic" in composition (sometimes a derivative product of andesite), meaning that it has } 67% SiO2, with important levels of Al2O3 (approx. 10%-15%), FeO and MgO (5%-10%, between them), CaO (2%), TiO2 ( {1%) and Na2O and K2O (5%-8%, between them).
These are sweeping generalizations, with data off the top of my head, but will serve you fairly well.
2. crystals, on the other hand, are usually not so robust in the sense that their compositions are more restricted, for example:
Na-K feldspars comprise Na, K, Al and Si, with minor Ca and very low Fe
Ca-Na feldspars comprise Ca, Na, Al and Si, with minor K......notably, Fe and Mg are not important for feldspar group minerals, whereas
Fe-Mg silicates of many silicate families comprise Fe, Mg, Ca, Al and Si, with the Al and Ca most variable, along with lesser Ti and Mn........notably, Na and K are not important for most common silicates (amphiboles are an exception)
clay minerals are mainly Al and Si and quartz, of course, is Si only (note: when geologists refer to any element in a mineral we invariably imply that it is associated with oxygen....thus Si in quartz is actually SiO2....we express all chemical analyses of rocks as oxides as oxygen is the most abundant element in Earth)
Whew! Geology 101 in an e-mail. Contact me if you have specific questions.
Winton
Dr. Winton Cornell Senior Research Associate & Supervisor, Microanalysis Laboratory Department of Geosciences The University of Tulsa 600 South College Tulsa, OK 74104-3189
Generally speaking high temp sources like volcanoes create spectacular spheres in a range of sizes but air sampling preferentially selects the midrange 0.5 to 10 micron with the larger setttling out and the smaller agglomerating. All this depends on atmospheric conditions. Wind blown dusts generally are all shapes and sizes, rarely spheres, with rounded edges from banging into each other. Composition in both cases is largely dependant on the source.
Greg Meeker (USGS) has a paper which discusses morphology and composition of volcanic plume particles - I highly recommend it:
Meeker, GP, Hinkley, TK "The Structure and Composition of Microspheres From the Kilauea Volcano, Hawaii" American Minerologist, Vol 78, Pg 873-876, 1993.
You may find the particle atlas usefull also.
Scott
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------------------------------ Scott Wight e-mail: SCOTT.WIGHT-at-NIST.GOV NIST - Microanalysis Group W voice: 301-975-3949 Bld 222, Rm A113 | fax:301-216-1134/301-417-1321 Gaithersburg, MD 20899 \|/ disclaimer: Any opinion expressed is my own and does not represent those of my employer.
My ancient paraffin embedding station recently died. I nearly suffered a heart attack when I saw the price of a replacement.
I am looking for a paraffin dispenser+hot plate+cold plate+forceps warmer. Although a work station is inviting because it has a trough system to collect all the drips, the $8,700 price that I was quoted is out of the question.
I have a separate embedding oven and vacuum embedding oven, so I do not need a cadillac model. I also am considering buying the units individually. Is there a hot plate or hot plate plus cold plate with a trough?
If you are a vendor (for either the work station or components), please contact me directly.
If you are a user/recent purchaser of any of the above, your comments would be appreciated.
Thank you.
Don
______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-tcnj.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 P.O. Box 7718 fax: (609) 637-5118 The College of New Jersey Ewing, NJ 08628-0718
We are looking for a tilt/rotate stage for our JEOL 2000FX. If you have one that you would like to dispose of, please contact me directly at: kkato-at-mail.arc.nasa.gov.
Katharine Kato
**************************************************************************** Katharine Kato SETI Institute 239-14 NASA Ames Research Center Moffett Field CA 94035-1000 ph# 650-604-5218 fax# 650-604-1088
In a message posted yesterday, the URL for a screenshot of the VIDX X-ray microanalysis system located on the Evex web site(s) was listed. Evex very quickly replaced the particular screenshot referred to in that posting with another one soon afterwards. The replacement screenshot has exactly the same URL as before. This coincidence in timing was extremely fortunate all around because the few curious enough to take a look at the original one might have been deceived into thinking that the Evex VIDX system looked very similar to their competitor's Win EDS system. Fortunately very few would have had the potential chance to be so deceived.
The screenshot in question has been replaced in a very timely fashion and the URLs mentioned in yesterday's message now draw point to a newer screenshot of the Evex "VIDX X-ray Microanalysis System" on the various web sites owned by Evex. These URLs are for example
http://www.thomson-scientific.com/anal.htm and http://www.evex.com/anal.htm etc etc
The screenshots of the competing Thomson Scientific WinEDS Microanalysis System on the site owned by the long-established Australian company, Thomson Scientific Instruments Pty Ltd, at
http://werple.net.au/~tsi/brochure.htm
remain unchanged. The recently updated image on the Evex sites makes it clearer that the VIDX system, in my opinion, no longer bears such a strong apparent similarity to the WinEDS system after all. The updated VIDX image suggests that there is at least one additional feature since yesterday, and since yesterday the system now apparently sports a significantly different user interface involving the transposition of two pairs of control buttons. The revised image seems to be a more accurate representation since some previously incorrect peak identification labels on the EDS spectrum have also been corrected. Any remaining apparent visual similarities to WinEDS, such as the graphics on the control buttons and elsewhere that are fairly distinctive and complex, but not necessarily intentionally identical since they could be derived independently, could be entirely coincidental. This is particularly the the case now, since the current VIDX screenshot image has an even lower displayed resolution than yesterday's version. There is now an even less likely chance that potential customers could be accidentally deceived into thinking that there was any visual similarity between the two competing X-ray analysis systems. We should all thank Evex for making this especially timely effort to improve the perceived credibility of the information on their website(s).
Apologies for cluttering the list further with this topic but in the name of ethical business practices and fair competition, there is an obligation to make sure that Evex's very timely and commendable efforts to reduce any possible misunderstandings should not go unnoticed.
I shall now immediately cease any further participation in discussions on this or other Evex-related topics. It is not my intention to deceive anyone...
} } November 17, 1997 } } } } It is not Evex Analytical's intention to deceive anyone. } } } } Promotion of Evex Analytical Inc., is promoted only at } } www.evex.com } } } } Evex Analytical does own many other websites but promotes its products at } } www.evex.com } } } } http://www.evex.com/971117.htm } } There is more.... } } In the context of the preceeding messages in this thread, the curious might } want to take a look at screenshots of the Evex "VIDX X-ray Microanalysis } System" that can be viewed at either of the following identical websites, } both "owned" by Evex: } } http://www.evex.com/anal.htm } } and } } http://www.thomson-scientific.com/anal.htm } } } and the screenshot of the WinEDS Version 3.0 microanalysis system marketed } by the real (and I imagine the original) Thomson Scientific at the } following totally unrelated website: } } http://werple.net.au/~tsi/brochure.htm } } Not making any allegations here, but WHAT GIVES?? } Spooky stuff.
Alexandr G. Domantovski wrote: Hi, dear colleagues!
} For radial distribution function (RDF) method I need a } receipt to take coherent part of electron scattering curve. } Any suggestions or literature references (journals, } conferences, etc.-it is better) would be appreciated. } Thank you very much for attention to my problem. } } Alexsandr Domantovski. } RRC "Kurchatov institute", Russia.
I think, the classical papers on RDF calculations are: 1. J.Konnert and J. Karle, Acta Crystalographica A29 (1973) 702 (and other papers by J. Karle) It contains a very detailed procedure for optimization of the extraction of coherent part of the scattering. This procedure can be used in full or partially.
2. R.Kaplow et al. Phys.Rev. 138 (1965) A1336. A very useful paper describing the sources of errors in RDF calculation.
3. D.J.H.Cockayne, D.R.McKenzie Acta Crystal. A44 (1988) 870; and Phil. Mag. B54 (1986) 113. More recent papers giving a complete procedure and some underlying problems.
I'll appreciate any information you'll get from the listserver.
If you can't do ultracryotomy, why don't you take off the cell wall with enzymes (check out refs by Laura Davis and others; you might also treat the cytoplamsic membrane with detergents like saponin. Of course chewing your antibody to smaller bits would help too--but that's not my area, sorry.
Sara
On Wed, 19 Nov 1997 TIMOTHY.M.BOURETT-at-usa.dupont.com wrote:
} Date: Wed, 19 Nov 1997 09:40:18 -0500 (EST) } From: TIMOTHY.M.BOURETT-at-usa.dupont.com } To: microscopy-at-sparc5.microscopy.com } Subject: Making Fab Fragments with IgMs } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Greetings, } } We are doing whole-mount immunofluorescence of walled fungal cells, and } have run into problems of permeability of a IgM antibody we wish to label with. } We have no problems with our IgG antibodies, but this IgM simply does not get } in (at least not consistently and evenly). I was told that it may be possible } to digest the IgM with trypsin to generate Fab fragments. I was also told that } it is difficult to control the activity of the protease to prevent unwanted } cleaving. Does anyone have any experience with this approach? A reference } with a protocol would be greatly appreciated!! Likewise, any additional } advice/comments would be welcome. Thanks. } } Tim Bourett } DuPont Experimental Station } Wilmington, DE USA }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
On Mon, 17 Nov 1997 hefeh-at-Rcs1.urz.tu-dresden.de wrote:
} Date: Mon, 17 Nov 1997 12:57:23 +0100 } From: hefeh-at-Rcs1.urz.tu-dresden.de } To: Microscopy-at-sparc5.microscopy.com } Subject: ImmunoLM - CD4/CD8-antibodies } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello everybody ! } } We are looking for antibodies against CD4 and CD8 that can be used with } formalin fixed, paraffin embedded rat tissues - with or without application } of antigen retrieval procedures. } Has anybody some helpful experience ? } } Thanks alot. } } Heinz } } **************************************************************************** } Dr. Heinz Fehrenbach } Institute of Pathology } University Clinics "Carl Gustav Carus" } Technical University of Dresden } } Fetscherstr. 74 Phone: ++49-351-458-5277 } D-01307 Dresden Fax: ++49-351-458-4328 } Germany e-mail: hefeh-at-rcs.urz.tu-dresden.de } **************************************************************************** } }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
I search standards for an electron-microprobe. We will analyse different ceramic samples.
My adress: Ewald Eipper Institute for Mineralogy, Petrology and Geochemistry Universty of Tuebingen Wilhelmstr. 56 72074 Tuebingen my e-mail: ewald.eipper-at-uni-tuebingen.de
Some recent work showing RDF's for amorphous Pd-Si using electron diffraction is in
M. Matsushita, Y. Hirotsu, K. Anazawa, T. Ohkubo and T. Oikawa, "Electron diffraction intensity analysis of amorphous Pd75Si25 alloy thin films with imaging-plate technique", Materials Transactions JIM vol 36 (1995) pp 822-827
The authors correct for inelastically scattered intensity based on EELS measurements. The figures in the article plot both total and elastic intensity as functions of Q, which may be of some help. I am not sure if there is more recent work along these lines, but a search of some of these authors might provide additional material.
Wharton Sinkler
On Wed, 19 Nov 1997, Serge Oktyabrsky wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Alexandr G. Domantovski wrote: } Hi, dear colleagues! } } } For radial distribution function (RDF) method I need a } } receipt to take coherent part of electron scattering curve. } } Any suggestions or literature references (journals, } } conferences, etc.-it is better) would be appreciated. } } Thank you very much for attention to my problem. } } } } Alexsandr Domantovski. } } RRC "Kurchatov institute", Russia. } } I think, the classical papers on RDF calculations are: } 1. J.Konnert and J. Karle, Acta Crystalographica A29 (1973) 702 (and other } papers by J. Karle) It contains a very detailed procedure for optimization } of the extraction of coherent part of the scattering. This procedure can be } used in full or partially. } } 2. R.Kaplow et al. Phys.Rev. 138 (1965) A1336. A very useful paper } describing the sources of errors in RDF calculation. } } 3. D.J.H.Cockayne, D.R.McKenzie Acta Crystal. A44 (1988) 870; and Phil. Mag. } B54 (1986) 113. More recent papers giving a complete procedure and some } underlying problems. } } I'll appreciate any information you'll get from the listserver. } } ***************************** } Dr. Serge Oktyabrsky } Visiting Scientist } phone 919-515-1104 } fax 919-515-7642 } e-mail: serge_oktyabrsky-at-ncsu.edu } ***************************** } } } } } }
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Wharton Sinkler PhD Department of Materials Science and Engineering Northwestern University 2225 North Campus Drive Evanston, IL 60208-3108 tel: (847) 491-7809 fax: (847) 491-7820 email: sinkler-at-apollo.numis.nwu.edu
We are examining doing some cryo-SEM on biohazardous material and have some concerns regarding the sterilization of equipment after use. Once the samples are in the cryo-prep unit and fractured for coating, the excess sample will remain in the unit and of course will eventually melt and be deposited on chamber components. There is also the potential for small fractions of the material to break off and enter into the vacuum system and be vented out.? Does anyone have experience in this area and can make some recommendations? Greatly appreciated.
Wayne England ORTECH CORP. wengland-at-ortech.on.ca
I would like to thank everyone for responding to my urgent need for parts for the ongoing installation of a Philips EM 300 at my institution. I have a number of leads on the required parts which I am currently following up with.
As you all know, this listserver is quite a valuable resource. Nestor is to be applauded for his efforts in keeping this 'online'. I sent off my original message at approximately 10:00pm (U.S. Eastern time) and couldn't begin to imagine that early the next morning I had five responses and later a phone call, with many offers to donate the needed parts.
Thanks again to all!
Steve
Stephen J. Beck Associate Professor Bio-Imaging Center/Electron Microscopy Department of Biology Nassau Community College Garden City, NY 11530 Voice Mail: (516) 572-7829 Email: {becks-at-sunynassau.edu} URL: {http://www.sunynassau.edu/webpages/biology/becks.htm}
I am looking for the controller for the Microspec WDX-2A Wavelength Dispersive Spectrometer. Any information on the subject will be greatly appreciated. Thank you very much in advance.
We are setting up a multi-user imaging facility at the=20 Cross Cancer Institute in Edmonton, and are looking for a=20 qualified person to manage it. Please pass this posting=20 along to anyone who might be interested. I would happy to=20 answer any questions. (Please direct them to=20 andrewsh-at-cancerboard.ab.ca. and not the list!!)
Thanks,
Andrew Shaw, Ph.D.
Advertisement for the POSITION OF MANAGER: Imaging Facility
Applications are invited for the position of Manager of a=20 new Imaging Facility located within the Division of =20 Experimental Oncology at the Cross Cancer Institute in=20 Edmonton. The Facility comprises a new Zeiss LSM 510 =20 CONFOCAL microscope, a Hitachi H7000 electron microscope,=20 and two Olympus fluorescence microscopes equipped with=20 EMPIX video image software for analysis of cell migration=20 and ion flux. The Cross Cancer Institute is the northern=20 Alberta comprehensive cancer treatment and research=20 facility of the Alberta Cancer Board and is situated=20 immediately adjacent to the University of Alberta campus.=20 Its academic staff are members of the=20 Department of Oncology of the Faculty of Medicine at the=20 University of Alberta. The Department of Oncology has more=20 than 80 members and six Divisions: Experimental Oncology,=20 Medical Oncology, Medical Physics, Palliative Medicine,=20 Radiation Oncology, and Surgical Oncology.=20
Research take place on four floors of new research space,=20 and includes DNA damage and repair, growth and spread of=20 malignancies, tissue injury, gene regulation and neoplasia,=20 tumor suppressor genes, and membrane transporters. Active=20 translational research programs are established in=20 neuro-oncology, pediatric oncology, breast cancer, prostate=20 cancer, hematologic malignancies, and gynecologic=20 malignancies. Further information on the research programs=20 of Experimental Oncology can be found at the Department of=20 Oncology=92s website (http://www.ualberta.ca/~oncology).
Applicants should have a Ph.D., or M.Sc. with 3-4 years=20 research experience in fluorescence microscopy, confocal=20 microscopy and preferably in the operation of an electron=20 microscope. Responsibilities will include the maintenance=20 of microscopes, computers and imaging software; the=20 instruction of users: and collaborative assistance with=20 research projects. This individual must be able to interact=20 and communicate effectively with scientists, trainees and=20 technical staff. As well, the individual will be expected=20 to provide advice to users on experimental design and to=20 assist users in preparing data for publication. The=20 manager will report to the academic supervisor of the=20 facility and will be a member of the Imaging Facility Users=20 Committee.
The University of Alberta was established in 1906, and is=20 today one of Canada=92s largest universities with a student=20 enrollment of over 28,000. More than 4,000 of these=20 students are graduate students registered in over 76=20 master=92s programs, and 64 doctoral programs. Edmonton, a=20 modern and progressive city of 750,000, is renowned for the=20 diversity and quality of its cultural and recreational=20 opportunities and is close to skiing and hiking in the=20 Canadian Rockies.
The Cross Cancer Institute is a smoke-free workplace. In=20 accordance with Canadian immigration requirements, priority=20 will be given to Canadian citizens and permanent residents=20 of Canada, but others are strongly encouraged to apply. =20 Salaries and benefits are competitive. Applicants should=20 submit their curriculum vitae and a list of three referees=20 by January 1, 1998 to Dr. Carol E. Cass, Chair of Oncology=20 and Associate Director (Research), Cross Cancer Institute,=20 University of Alberta, Edmonton, Alberta, T6G 1Z2.
Andrew R.E. Shaw, Ph.D. Associate Professor Oncology
---------------------- Andrew Shaw andrewsh-at-cancerboard.ab.ca
Dear microscopists: Is there anyone knowing how to keep aorta tissue stay on the glass microscopic slide? I have tried poly-L-lysion caoted and SectionLock (positive charged) slides for either immunohistochemistry and in situ hybridazation (sometimes both on the same slide). I don't know how to avoid tissue falling off. Thanks in advance.
Dorothy Zhang Harvard School of public Health Building 2, CVLAB 677 Huntington Ave, Boston, MA 02115 Phone# 617-432-2970 Fax#: 617-432-2980
I have a vial of copper TEM grids that are very hydrophobic. Does anyone know how I can make them wettable? Acid rinse? Alcohol rinse? Detergent rinse? Buy new grids?
TIA
Bob
Dr. Robert R. Wise Department of Biology and Microbiology University of Wisconsin-Oshkosh Oshkosh, WI 54901
(920) 424-3404 tel (920) 424-1101 fax wise-at-uwosh.edu www.uwosh.edu/departments/biology/wise/wise.html
FWIW... Once upon a time, Kevex also produced a control system for the '2A. Was called "Sesame". Horrible firmware bugs were never fixed, but it is usable once you become familiar with the special "features". Quant was handled, however, by the EDS processor. Woody
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am looking for the controller for the Microspec WDX-2A Wavelength Dispersive Spectrometer. Any information on the subject will be greatly appreciated. Thank you very much in advance.
On Thu, 20 Nov 1997 wise-at-vaxa.cis.uwosh.edu wrote:
} I have a vial of copper TEM grids that are very hydrophobic. Does anyone } know how I can make them wettable? Acid rinse? Alcohol rinse? Detergent } rinse? Buy new grids?
I barbecue my grids just before I pick up sections. Pass them several times quickly over the flame from an alcohol lamp or disposable lighter. With a little practice, you'll have clean, hydrophilic grids with no melted bars! Even with slim bar grids.
Aloha, Tina
http://www.pbrc.hawaii.edu/bemf/microangela/ **************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
} I have a vial of copper TEM grids that are very hydrophobic. Does anyone } know how I can make them wettable? Acid rinse? Alcohol rinse? Detergent } rinse? Buy new grids?
Several ways to skin this cat.
If using uncoated grids, when ready to pick up sections, simply pass the grid through an alcohol (not gas) flame to cherry color (almost instantaneously). This may slightly discolor the grid but it is now extremely hydrophilic. This takes some practice since most people tend to overheat and damage the grids but it is very convenient.
Individual grids may also be cleaned as needed by dipping (5-10x) into 4% nitric acid and then dipping several times in distilled water.
Groups of grids may be cleaned by swirling in 4% nitric acid for several minutes and rinsing in distilled water. Then rinse in ethanol and dry in an oven.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
We have a 230 MB magneto optical drive on a Noran V-III EDX system and need a second drive in order to use the disks on a Macintosh. Pinnacle Micro no longer makes the 230 MB Tahoe, so we are looking for another manufacturer. Anyone care to recommend a 230 MB model MO and vendor to purchase this device? Thanks.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
I have a vial of copper TEM grids that are very hydrophobic. Does anyone know how I can make them wettable? Acid rinse? Alcohol rinse? Detergent rinse? Buy new grids? TIA Bob
Holding each grid in tweezers, dip it into the mouth of a bottle containing concentrated nitric acid, for about 1 second. The fumes will render the grid hydrophilic without tarnishing the copper.
Ray Egerton, Physics Dept, University of Alberta, Edmonton, Canada T6G 2J1 Phone: 403-492-5095, FAX: 403-492-0714, e-mail: egerton-at-phys.ualberta.ca ------------------------------------------------------------------------
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } We have a 230 MB magneto optical drive on a Noran V-III EDX system and need } a second drive in order to use the disks on a Macintosh. Pinnacle Micro no } longer makes the 230 MB Tahoe, so we are looking for another manufacturer. } Anyone care to recommend a 230 MB model MO and vendor to purchase this } device? Thanks. } } #################################################################### } John J. Bozzola, Ph.D., Director } Center for Electron Microscopy } Neckers Building, Room 146 - B Wing } Southern Illinois University } Carbondale, IL 62901 } U.S.A. } Phone: 618-453-3730 } Fax: 618-453-2665 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/departments/shops/cem.html } #################################################################### } } }
At 13:01 20/11/97 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We project to heat metal alloys up to about 1200 C under mechanical constraints in order to observe phase transformation in situ in a MEB. In such experimental conditions (heat and light emitted from the sample), I guess there will be many problems with the detectors (Everhart-Thornley, BSE, EDX, EBSP). I ask me if, for this goal, there would be some advantages to work with an environnemental MEB (ESEM - gaseous secondary electron detector).
Any kind of experience in this field would of great interest for us. Many thanks for your answers.
Monique repoux-at-cemef.cma.fr
----------------------------------------------------------------------- Monique Repoux Tel: 33 (0)4 93 95 74 13 CEMEF 33 (0)4 93 95 75 91 Ecole des Mines de Paris B.P. 207 Fax: 33 (0)4 93 65 43 04 06904 SOPHIA ANTIPOLIS CEDEX e-mail: repoux-at-cemef.cma.fr FRANCE -----------------------------------------------------------------------
It has been a few years since I have been involved with temperature control stages on optical microscopes. Last I remember there were issues such as surface area temperature mapping(variations) functions, isolation, target range overshooting, steady state vs. drift functions, heating and cooling techniques, depth of focus and other optical issues, maximum and minimum feasible temperatures, heating and cooling techniques, etc. Can anyone refresh my memory , bring me up to date ,and refer me to journal and review articles on the subject. I would like references to current manufacturers. Also I would be interested in personal experiences with specific current equipment manufacturers of both microscopes and control stages for this use (preferably any potentially negative feedback opinions sent only to my personal email address below, so no manufacturer is embarrassed or damaged by personal opinions in this forum/possibly an obvious point of controversy). Finally does anyone have any personal experience with modifications to existing designs or references to in house construction of these devices. I also might be prospect for use equipment of this nature.
JD EMAIL1: wa5ekh-at-juno.com please cc to EMAIL2: wa5ekh-at-cyberramp.net ------=_NextPart_000_01BCF62F.139292C0 Content-Type: text/html; charset=ISO-8859-1 Content-Transfer-Encoding: base64
The experience I had while performing the in-situ experiments on the hot-filament CVD growth of diamond films in the ESEM showed, that in case of Danilatos's detector (ESD for ESEM) the major problem at high temperatures is the thermal electron emission. If your material would not have a high thermal electron emission at this temperature, you can succeed. But the major problems with the metals is their ability to emmit electrons at high temperature. The low energy of thermal electrons does not matter, thay will be accelerated by the EDS detector. If you will be able to supress somhow the thermal electron emission, ESEM (ESD detector) may be extremely helpfull. Possible solution is to apply some positive voltage to the sample (the voltage should be slighly higher than the exit work).
Best regards and a good luck. If you have any other questions about ESEM in-situ experiments you are welcome.
Nick Kinaev (The university of Queensland, Australia)
} We project to heat metal alloys up to about 1200 C under mechanical } constraints in order to observe phase transformation in situ in a MEB. } In such experimental conditions (heat and light emitted from the sample), I } guess there will be many problems with the detectors (Everhart-Thornley, } BSE, EDX, EBSP). } I ask me if, for this goal, there would be some advantages to work with an } environnemental MEB (ESEM - gaseous secondary electron detector). } } Any kind of experience in this field would of great interest for us. } Many thanks for your answers. } } Monique } repoux-at-cemef.cma.fr } } ----------------------------------------------------------------------- } Monique Repoux Tel: 33 (0)4 93 95 74 13 } CEMEF 33 (0)4 93 95 75 91 } Ecole des Mines de Paris } B.P. 207 Fax: 33 (0)4 93 65 43 04 } 06904 SOPHIA ANTIPOLIS CEDEX e-mail: repoux-at-cemef.cma.fr } FRANCE } ----------------------------------------------------------------------- } } } }
Nick Kinaev; Centre for Electron Microscopy(CMM) The University of Queensland E-Mail : nick-at-mama.minmet.uq.oz.au _ . ,~' (_|\ ,-' \ Ph. home : +61 7 3279 4771 ( * {----Brisbane Ph. Dept:+61 7 3365 3743 \ __ / Qld 4072 Fax: +61 7 365 3888 \,~' "\__ / Australia
Monique The ESEM secondary electron detector, ESD in the older instruments and GSED in the newer instruments, is not light sensitive as the other detectors (E-T, BSD, and light element EDS) are very light sensitive. The ESEM has a special high temp ESD for use with the hot stage and I believe the heat shield is recommened for that temp. I would also take care to protect your EDS window from not only the light but also heat (IR) and projectiles. Good luck, Scott Wight
} } We project to heat metal alloys up to about 1200 C under mechanical } constraints in order to observe phase transformation in situ in a MEB. } In such experimental conditions (heat and light emitted from the sample), I } guess there will be many problems with the detectors (Everhart-Thornley, } BSE, EDX, EBSP). } I ask me if, for this goal, there would be some advantages to work with an } environnemental MEB (ESEM - gaseous secondary electron detector). } } Any kind of experience in this field would of great interest for us. } Many thanks for your answers. } } Monique } repoux-at-cemef.cma.fr } } ----------------------------------------------------------------------- } Monique Repoux Tel: 33 (0)4 93 95 74 13 } CEMEF 33 (0)4 93 95 75 91 } Ecole des Mines de Paris } B.P. 207 Fax: 33 (0)4 93 65 43 04 } 06904 SOPHIA ANTIPOLIS CEDEX e-mail: repoux-at-cemef.cma.fr } FRANCE } -----------------------------------------------------------------------
------------------------------------------------------------------ Scott Wight e-mail: SCOTT.WIGHT-at-NIST.GOV NIST - Microanalysis Group W voice: 301-975-3949 Bld 222, Rm A113 | fax:301-216-1134/301-417-1321 Gaithersburg, MD 20899 \|/ disclaimer: Any opinion expressed is my own and does not represent those of my employer.
} I have a vial of copper TEM grids that are very hydrophobic. Does anyone } know how I can make them wettable? Acid rinse? Alcohol rinse? Detergent } rinse? Buy new grids? } Dear Bob, I put the grids in a plasma cleaner for ~1 min; this may be more difficult than the other methods suggested. Yours, Bill Tivol
} I have a vial of copper TEM grids that are very hydrophobic. Does anyone } know how I can make them wettable? Acid rinse? Alcohol rinse? Detergent } rinse? Buy new grids? } Dear Bob, I put the grids in a plasma cleaner for ~1 min; this may be more difficult than the other methods suggested. Yours, Bill Tivol
} I have a vial of copper TEM grids that are very hydrophobic. Does anyone } know how I can make them wettable? Acid rinse? Alcohol rinse? Detergent } rinse? Buy new grids? } Dear Bob, I put the grids in a plasma cleaner for ~1 min; this may be more difficult than the other methods suggested. Yours, Bill Tivol
Hello Friends, I would be interested in hearing opinions about the features that microscopists consider essential on a new purchase of a confocal microscope for biological work.
Which scopes do you consider worthy, user friendly, cost efficient and the best quality optically ? Are there any features or scopes that have proved, in use, not to perform up to expectations ? I would welcome input from users and vendors. Thanks,
Linda M. Fox lfox1-at-wpo.it.luc.edu Loyola University Medical Center Dept. of CBN and Anatomy 2160 S. First Ave. Maywood, Illinois 60153 1-708-216-3395 or Dr. John A. McNulty 1-708-216-5161
I have not used them so I don't know if I can recommend them but APS Technologies makes a 230 MB MO drive that says can be used on Mac, PC or NT systems. Costs are between $300 and $400 depending on the one you get. Their phone number is 800-766-8427 and is available 7 days/wk. 24 hrs./day. They also sell many other drives and accessories including hard drives CD ROM and writeable CD drives.
I have no interest in APS or their products.
Michael D. Standing BYU Microscopy Lab e-mail: Michael_Standing-at-byu.edu -----Original Message-----
Some years ago I did some observations of crystalization in waste glass at about 1000 C through the generosity of Electroscan Corporation which had developed the ESEM (It has now gone over to Philips). I also did some additional observations at 700-800 C of crystallization in other glasses. I has absolutely no trouble making the observations. I would presume the detector would work well in your application as well. I have no personal interest in Philips/Electroscan. This is just my personal experience. Andy Buechele The Catholic University of America 409 Hannan Hall Washington, D.C. 20064 (202) 319-4995 FAX: (202) 319-4469
Bob, A lot of good suggestions, but I keep a stock of 3% HCl in 95% Et-OH on hand for this purpose. In bulk, I swirl them in a small amount of the cleaning solution for ~30 sec., rinse several times with DH2O and dry them using a Buchner funnel with a piece of filter in the bottom. They remain hydrophylic for quite a while (days,weeks?) before they need to be treated again. Individually, they may be dipped 2-3 times in the cleaning solution, dipped several times in DH2O, dried on filter paper and used. Bill wise-at-vaxa.cis.uwosh.edu wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I have a vial of copper TEM grids that are very hydrophobic. Does anyone } know how I can make them wettable? Acid rinse? Alcohol rinse? Detergent } rinse? Buy new grids? } } TIA } } Bob } } Dr. Robert R. Wise } Department of Biology and Microbiology } University of Wisconsin-Oshkosh } Oshkosh, WI 54901 } } (920) 424-3404 tel } (920) 424-1101 fax } wise-at-uwosh.edu } www.uwosh.edu/departments/biology/wise/wise.html
-- ============================================================= Bill Chissoe III Electron Microscopist,University of Oklahoma E-mail: wchiss-at-ou.edu Ph. (405)325-4391 =============================================================
Our 230Meg Optical drive is a Fujitsu Dynamo, bought about two years ago from one of the large computer places (Insight, PC Warehouse, etc., etc - I don't remember which specific one now). I've no idea, though, if it is still available.
Tony Garratt-Reed
At 05:04 AM 11/21/97 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Anthony J. Garratt-Reed MIT Room 13-1027 77 Massachusetts Avenue Cambridge, MA 02139-4307 United States of America
} I was interested to see the various responses to this question, as my own solution has been to wash the grids in fairly concentrated NaOH for a few seconds, followed by several distilled water rinses. } } Tony Garratt-Reed } } At 01:01 PM 11/20/97 -0600, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Anthony J. Garratt-Reed MIT Room 13-1027 77 Massachusetts Avenue Cambridge, MA 02139-4307 United States of America
Responding to the message of {3.0.3.32.19971121203127.006b0328-at-mama.minmet.uq.oz.au} from Nikolai Kinaev {nick-at-mama.minmet.uq.oz.au} : } [...] } } The experience I had while performing the in-situ experiments on the } hot-filament CVD growth of diamond films in the ESEM showed, that in case } of Danilatos's detector (ESD for ESEM) the major problem at high } temperatures is the thermal electron emission. If your material would not } have a high thermal electron emission at this temperature, you can succeed. } But the major problems with the metals is their ability to emmit electrons } at high temperature. The low energy of thermal electrons does not matter, } thay will be accelerated by the EDS detector. If you will be able to } supress somhow the thermal electron emission, ESEM (ESD detector) may be } extremely helpfull. Possible solution is to apply some positive voltage to } the sample (the voltage should be slighly higher than the exit work). } } Best regards and a good luck. } If you have any other questions about ESEM in-situ experiments you are } welcome. }
I believe this is what has been done for the ESEM 1500 degree hot stage. We have used the regular hot stage (1100 degrees max), and a prototype of the high temperature one at the Electroscan factory to look at melting of films on ceramic substrates. As I remember, 1100 - 1200 is the regime where it became crucial to suppress the thermal electrons in order to get a useable image with the GSED.
__________________ Stuart McKernan stuartm-at-tc.umn.edu Microscopy Specialist CIE Characterization Facility, University of Minnesota Phone: (612) 626-7594 100 Union Street S. E., Minneapolis, MN 55455 Lab: (612) 624-6590
I need to find replacement bulbs for a 1977 Leitz HM-LUX light microscope. Where do I find bulbs to replace the one that burned out. Is there a one stop light bulb source? Thanks for any information you might send my way.
I'll see you later,
Paula = )
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu
At 09:43 AM 11/21/1997 -0600, you wrote: } I would be interested in hearing opinions about the features } that microscopists consider essential on a new purchase of a confocal } microscope for biological work.
Our lab is also thinking of purchasing a confocal. I'd love to be included in this discussion if it goes to private mail. -=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-= Paul Tiseo | "It takes a big man to cry... Mayo Clinic - Jacksonville | but it takes a bigger man Birdsall 3 | to laugh at that man." (904) 953-8254 (pager) | tiseo.paul-at-mayo.edu | http:// coming soon | - Jack Handey -=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=
I find the following company a great source of low cost bulbs. I pay less than 1/2 of what I had been paying at other sources. One of my microscope dealers told me the last bulb I got was for less than he gets them for! They give quotes over e-mail.
------------------------------------------- Janet Connolly Lamp Technology, Inc. lamptech-at-panix.com 1-800-KEEP-LIT "We a have bulb for every socket" http://www.webscope.com/lamptech/ -------------------------------------------
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (573)-882-4712 (voice) (573)-882-0123 (fax)
I have ~3 years of the journal Microscopy Research & Technique, It's current; altho I resigned as an editor a year ago, I've continued to receive them. I'll send them to someone who is willing to pay UPS shipping costs.
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
Boy, did this one fill up my in box. Thanks for all the replies. We're going to try the alcohol lamp first and see how that works for us.
Thanks again
Bob Wise
**************
I barbecue my grids just before I pick up sections. Pass them several times quickly over the flame from an alcohol lamp or disposable lighter. With a little practice, you'll have clean, hydrophilic grids with no melted bars! Even with slim bar grids.
Tina (Weatherby) Carvalho
****************
Wash in 100% EtOH, maybe followed by 100% Acetone. Or Ac 1st, then EtOH, doesn't matter. If really bad, do a wash with distilled water and dilute mild detergent first. To be honest, I forget if I sonicated or just swirled (been a while since I had to do this). Dry thoroughly.
When I've had this problem, it was because of an organic film that collected on the grids, usually air borne crud.
Phil Oshel
***************
I usually rinse them in acetone before I use them. This cleans the oils off of them.
Jane Glamp
***************
If using uncoated grids, when ready to pick up sections, simply pass the grid through an alcohol (not gas) flame to cherry color (almost instantaneously). This may slightly discolor the grid but it is now extremely hydrophilic. This takes some practice since most people tend to overheat and damage the grids but it is very convenient.
Individual grids may also be cleaned as needed by dipping (5-10x) into 4% nitric acid and then dipping several times in distilled water.
Groups of grids may be cleaned by swirling in 4% nitric acid for several minutes and rinsing in distilled water. Then rinse in ethanol and dry in an oven.
John J. Bozzola
*********************** "etch" them in 1n HCl for a few minutes (they will turn shiny!), rinse in dist. water and dry from ethanol. They will turn hydrophobic in a few days, a pain. My trick is to pull the grid through an alcohol flame (next to the microtome) just befor pick up. They will colorize (oxidise), but I never had a problem with contamination etc. It's a dream to pick up the sections!
Markus F. Meyenhofer
*************** Holding each grid in tweezers, dip it into the mouth of a bottle containing concentrated nitric acid, for about 1 second. The fumes will render the grid hydrophilic without tarnishing the copper.
Ray Egerton **************** A real easy way to make them hydrophilic is to use a small flame from e.g. an ethanol spirit lamp. Whisk each grid through the flame so it flashes red. Too long and its oxide! Makes them quite hydrophilic.
Melvyn Dickson ****************** what works best: try to 'glow-discharge' them. These units can be made in a workshop, but you also can buy them, are easy to use, and work reliably. (you can get the address of a supplier on request)
What may work (and can be tried instantly): rinse in ~ 10% sulfuric acid briefly, or for a 1 or 2 minutes, then in water, finally in acetone (pure) for a few min. (the latter treatment is used in our lab to clean gold carriers for freeze-etching; with copper grids, it might be a problem ...)
Reinhard Rachel *************** I would suspect that they have some type of an oil coating to (presumably) prevent oxidation, in which case a sonication in acetone should do the trick.
Brian G. Demczyk *************** I put the grids in a plasma cleaner for ~1 min; this may be more difficult than the other methods suggested.
Bill Tivol *************** My own solution has been to wash the grids in fairly concentrated NaOH for a few seconds, followed by several distilled water rinses.
Tony Garratt-Reed *************** I keep a stock of 3% HCl in 95% Et-OH on hand for this purpose. In bulk, I swirl them in a small amount of the cleaning solution for ~30 sec., rinse several times with DH2O and dry them using a Buchner funnel with a piece of filter in the bottom. They remain hydrophylic for quite a while (days,weeks?) before they need to be treated again. Individually, they may be dipped 2-3 times in the cleaning solution, dipped several times in DH2O, dried on filter paper and used.
Bill Chissoe *************** We clean our grids by dipping them into 0.1N HCL(or 1.0N HCL, if you want them really clean) for 10 times; followed by few dips in 95%ETOH; followed by few dips in acetone. You can use this technique with a few grids one at a time or you can clean an entire vial of 100. When cleaning grids in a vial, you can pipette the solutions into and out of the vial, cap the vial and shake between solutions..
Brendalyn Bradley-Kerr ************** I usually just dip my grids in a 0.1N solution of HCl just prior to picking up sections...that's it. I don't rinse them or anything!
Beth Fischer
Dr. Robert R. Wise Department of Biology and Microbiology University of Wisconsin-Oshkosh Oshkosh, WI 54901
(920) 424-3404 tel (920) 424-1101 fax wise-at-uwosh.edu www.uwosh.edu/departments/biology/wise/wise.html
1) Does anyone know where to get special order reticles and graticles? My microscope supplier does not have what I need.
2) I also need to know where to buy a counter (I have no idea what the correct name is!) that allows you to keep track of the number of intersections you count in a field placement. These are small mechanical devices with a button that increases the counter.
Thanks in advance,
Robin Griffin UAB Materials and Mechanical Engineering
At 09:16 21/11/97 +0001, you wrote: } I would advance that the reason that flaming tghe grid renders it } hydrophillic is that the surface is roughened (either small distortions } in the grid during heating or the presence of carbonaceous residue on } the surface), which may be undesirable in the long run as this surface } would be prone to pick up contaminants that may subsequently outgas in the } microscope. Perhaps treatment in a non-destructive (to the grid) solvent } would be preferable.
I always thought that flaming burned off a thin film of organic matter on the grids. They dont distort visibly on flaming. Neither do they turn black from carbonaceous reside, as the alcohol flame burns cleanly. But they change colour with formation of a copper oxide film.
As for the risk of picking up contaminants they are already contaminated with the material which makes them hydrophobic. Also the time elapsed between flaming and insertion into the microscope is only about an hour as staining and washing proceeds so there is small opportunity for contamination.
If you want guaranteed good adhesion to flamed grids, you should deliberately "contaminate" them with "scotch"tape adhesive dissolved on chloroform.
I suspect any organic solvent will leave a trace of organic matter so avoid them where I can.
Mel Dickson Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
I like to hear of your experience (if any) when placed into a similar situation.
For more than 40 years I have been polishing specimens for metallographic and geological examinations by optical and electron microscopical methods. Various standard techniques were used in the past and are still being applied; in addition, I teach my students these techniques. In may circumstances I also use diamond paste with ethanol as lubricant.
However, one of our safety officers suddenly 'discovered' that ethanol has a carcinogenic classification. As a result he wants all polishing with ethanol stopped unless we polish inside a fume cupboard and wear gloves.
He even suggested that we issue breathing masks to my students before they can polish samples using ethanol.
How would you react to a similar situation ?
Hans Brinkies Senior Lecturer SWINBURNE, University of Technology Industrial Microscopy HAWTHORN, Vic. 3122 - Australia hbrinkies-at-swin.edu.au
} ... } } However, one of our safety officers suddenly 'discovered' that } ethanol has a carcinogenic classification. As a result he wants all } polishing with ethanol stopped unless we polish inside a fume } cupboard and wear gloves. } } ...
Excuse me??? ... grain alcohol ... a carcinogen??? This may be the case for 200proof (100%) ETOH, because I think they may use drying agents ... but I can't imagine this being true for 190proof (95% - 5%H2O) ETOH!!!
Let's straighten this out and get the facts ...
cheerios, shAf -- ~~~~~~~~~~~~~~~~~~~~~~~ cogito, ergo zZOooOM ~~~~~~~~~~~~~~~~~~~~ Michael Shaffer - Geological Sciences - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
} I like to hear of your experience (if any) when placed into a similar } situation. } } For more than 40 years I have been polishing specimens for } metallographic and geological examinations by optical and electron } microscopical methods. Various standard techniques were used in } the past and are still being applied; in addition, I teach my } students these techniques. In may circumstances I also use diamond } paste with ethanol as lubricant. } } However, one of our safety officers suddenly 'discovered' that } ethanol has a carcinogenic classification. As a result he wants all } polishing with ethanol stopped unless we polish inside a fume } cupboard and wear gloves. } } He even suggested that we issue breathing masks to my students before } they can polish samples using ethanol. } } How would you react to a similar situation ? } } } Hans Brinkies } Senior Lecturer } SWINBURNE, University of Technology } Industrial Microscopy } HAWTHORN, Vic. 3122 - Australia } hbrinkies-at-swin.edu.au
Philip Oshel PO Box 5037 Station A Champaign, IL 61825-5037 (217) 355-1143 oshel-at-ux1.cso.uiuc.edu or poshel-at-hotmail.com ***** looking for a job *****
Monique Repoux wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } We project to heat metal alloys up to about 1200 C under mechanical } constraints in order to observe phase transformation in situ in a MEB. } In such experimental conditions (heat and light emitted from the sample), I } guess there will be many problems with the detectors (Everhart-Thornley, } BSE, EDX, EBSP). } I ask me if, for this goal, there would be some advantages to work with an } environnemental MEB (ESEM - gaseous secondary electron detector). } } Any kind of experience in this field would of great interest for us. } Many thanks for your answers. } } Monique } repoux-at-cemef.cma.fr } } ----------------------------------------------------------------------- } Monique Repoux Tel: 33 (0)4 93 95 74 13 } CEMEF 33 (0)4 93 95 75 91 } Ecole des Mines de Paris } B.P. 207 Fax: 33 (0)4 93 65 43 04 } 06904 SOPHIA ANTIPOLIS CEDEX e-mail: repoux-at-cemef.cma.fr } FRANCE } ----------------------------------------------------------------------- Monique,
Is it necessary that you use electron microscopy? Years ago, when I worked for Cambridge Instruments (now Leica), we had a vacuum furnace which fit on an inverted metallograph for just such a purpose. It could handle temperatures up to 1800 degrees C and had gas inlet/outlets so that you could purge the chamber with inert gases to minimize secondary chemical reactions. The chamber, itself, was made by the Reichert part of the Cambridge family (Austria).
I don't know if it is still produced, but if it would be of use, you might be able to find one in Europe by contacting the Reichert factory in Vienna. Norbert Wacht is probably a good technical contact there.
Best of luck!
Barbara Foster Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108 USA PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com
DISCLAIMER: We do not sell or have any commercial interest in this device.
} I like to hear of your experience (if any) when placed into a similar } situation. } } For more than 40 years I have been polishing specimens for snips ... } polishing with ethanol stopped unless we polish inside a fume } cupboard and wear gloves. } } He even suggested that we issue breathing masks to my students before } they can polish samples using ethanol. } } How would you react to a similar situation ? } } } Hans Brinkies
As an experiment, you could try polishing with high proof vodka. The safety officer could hardly put that out of bounds:)
Regards,
-- Larry Stoter 17, Rocks Park Road, Uckfield, E. Sussex, TN22 2AT, UK email: LPS-at-teknesis.demon.co.uk Phone/Fax: +44 (0)1825 767967
That could be a double-edged sword - it is true, I guess, that heavy exposure by drinking does induce cirrhosis (I can't spell that at this time of the morning!) of the liver. This could be a long-term strategy for getting rid of safety officers! Also a 'life-time' of heavy exposure to the vapour may have the same effect. But come on, where's reality? I have had this with my opposite number here, who wants to preserve us (admirably) from the local hospice, yet he drinks a lot!
It is a question of degree and exposure. In the UK, under the Control of Substances Hazardous to Health Regulations 1994, the Occupational Exposure Standard is 1000 ppm (1920 mg per cu. m.) averaged over an 8 hour reference period. There is no short-term limit (10 minutes), implying that it is not a serious hazard. It is possible to buy a Draeger hand pump and relevant glass tubes containing an indicator to monitor the air-borne concentration, but I would think that common sense can prevail! Providing there is some ventilation, exposure should be no more than that experienced when taking a strong drink!
I need to measure the height of {10um volcanic ash particles. Can anyone recommend software (preferably shareware) that will accurately make these measurements from a SEM stereo pair? Also, references for relevant research papers would be appreciated, too.
TIA from Michigan's snowy Upper Peninsula.
Owen
============================= Owen P. Mills Michigan Technological University Metallurgical & Materials Engineering Rm 512 MME Building Houghton, MI 49931 906-487-2002 906-487-2934 FAX opmills-at-mtu.edu
I checked the MSDS sheet that I have for ethanol and under Carcinogen Status it says NONE. They may have changed that ruling lately but you can surf the web for MSDS sites and find one that is more up to date. Your safety officer may be concerned because it is a solvent. Good luck trying to straighten this one out.
Roberto Garcia Electron Microscopy Facility Manager Wright State University
On the more serious side, is it possible that your safety officer has ethanol and IPA mixed up? According to Fisher's MSDS sheets on line, this is what they say about Carcinogenicity of various alcohols:
Carcinogenicity: Ethyl Alcohol - Not listed by ACGIH, IARC, NIOSH, NTP, or OSHA. Methyl alcohol - Not listed by ACGIH, IARC, NIOSH, NTP, or OSHA. Isopropyl alcohol - IARC: Group 3 carcinogen
This is taken from the Fisher Scientific website at
http://www.fisher1.com/
I have no connection to fisher.
David
Hans Brinkies wrote:
hbrinkies-at-lucy.cc.swin.edu.au on 11/24/97 04:19:31 AM
To: microscopy-at-Sparc5.Microscopy.Com cc: (bcc: David Bell)
I like to hear of your experience (if any) when placed into a similar situation.
For more than 40 years I have been polishing specimens for metallographic and geological examinations by optical and electron microscopical methods. Various standard techniques were used in the past and are still being applied; in addition, I teach my students these techniques. In may circumstances I also use diamond paste with ethanol as lubricant.
However, one of our safety officers suddenly 'discovered' that ethanol has a carcinogenic classification. As a result he wants all polishing with ethanol stopped unless we polish inside a fume cupboard and wear gloves.
He even suggested that we issue breathing masks to my students before they can polish samples using ethanol.
How would you react to a similar situation ?
Hans Brinkies Senior Lecturer SWINBURNE, University of Technology Industrial Microscopy HAWTHORN, Vic. 3122 - Australia hbrinkies-at-swin.edu.au
I got ethanol from AlfaAesar one month ago. The MSDS says 'Not listed as carcirogen by OSHA, IARC or NTP'. And it's for the usual mixture of 90% of ethanol, 5% of methyl alcohol and 5% isopropil alcohol, which is sold under name of ethanol (ethil alcohol) and wich most of us are using for metallography, microscope cleaning (but not drinking). By the way, I have another question conserning ethanol. In which cases it's preferable to use real 96% ethanol (prohibited for unauthoriside sale by goverment) and not denaturated alcohol described above (not in biology)?
Vladimir Dusevich Senior Analyst North Carolina State University
I got ethanol from AlfaAesar one month ago. The MSDS says 'Not listed as carcirogen by OSHA, IARC or NTP'. And it's for the usual mixture of 90% of ethanol, 5% of methyl alcohol and 5% isopropil alcohol, which is sold under name of ethanol (ethil alcohol) and wich most of us are using for metallography, microscope cleaning (but not drinking). By the way, I have another question conserning ethanol. In which cases it's preferable to use real 96% ethanol (prohibited for unauthoriside sale by goverment) and not denaturated alcohol described above (not in biology)?
Vladimir Dusevich Senior Analyst North Carolina State University
You should be able to get assistance from MicroBrightField in Colchester VT. Their phone # is (802) 655 9360 and their email address is info-at-microbrightfield.com. Their web address is www.microbrightfield.com/.
On Sun, 23 Nov 1997 rgriffin-at-eng.uab.edu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } 1) Does anyone know where to get special order reticles and graticles? } My microscope supplier does not have what I need. } } 2) I also need to know where to buy a counter (I have no idea what the } correct name is!) that allows you to keep track of the number of } intersections you count in a field placement. These are small } mechanical devices with a button that increases the counter. } } Thanks in advance, } } Robin Griffin } UAB Materials and Mechanical Engineering }
Edmund Glaser, D. Eng. Dept. Physiol. Univ. Md. School. Med. Baltimore, MD 21201 USA Ph: (410) 706-5041 Fax: (410) 706-8341
According to my references, effective 5-18-96, ethanol listed on the ACGIH Carcinogenicity list. However, it's classification code is A4, or "not classifiable as a human carcinogen." I have no record of it being listed by any other agency (IARC, NTP, OSHA, NIOSH, or MAK). I of course can't vouch for the reliability of the data in my third party database.
Perhaps you should ask your safety officer WHO classified ethanol as carcinogenic, and check into the accuracy yourself.
Jim Passmore
---------- } From: hbrinkies } To: microscopy } Subject: Ethanol } Date: Monday, November 24, 1997 1:19AM } } { {File Attachment: ETHANOL.TXT} } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
A similar situation prevailed here for a while: we had to treat ethanol as a carcinogen because of certain legal requirements related to its being on an official list of carcinogens or suspect ones (National Toxicology Program). However, a glimmer of intelligence appeared somewhere, and it was decided (I use the passive because I have no idea who) that the carcinogenic activity was related to excessive ingestion rather than inhalation, and that the inhalation hazard was similar to other solvents. So it disappeared from the list.
We still have to keep it locked up.
My opinions... Leonard Corwin Fort Dodge Animal Health (Analytical Research) Princeton, NJ 08543-0400 corwinl-at-pt.cyanamid.com
} I need to measure the height of {10um volcanic ash particles. Can anyone } recommend software (preferably shareware) that will accurately make these } measurements from a SEM stereo pair? Also, references for relevant } research papers would be appreciated, too. } We have a program called STERECON which does precisely what you want. It is not shareware (& our version may run only on a mainframe). There should be many papers with Mike Marko &/or Karolyn Buttle which use this program, & I can ask them for a list of references. I'm not sure of the availability of STERECON, but it is always possible to use it at our place (we're a NIH-funded biotechnological resource). Yours, Bill Tivol
If you are interested in the only SINGLE CD-ROM containing complete US demographic data from 67 Census Bureau CDs, for less than $200, then please read on.
CensusCD contains the entire US Census, over 1.3 billion demographics, with an easy to use yet powerful Windows interface, all on one(1) CD-ROM. It is the best reference for researching the people and housing of the United States.
CensusCD gives you access to demographics on population(2633 variables), housing(714), families(385), employment(301), occupation(16), education(155), college(19), income(862), White(250), African American(249), Asian(296), Hispanic(292), Native American(267), language spoken(26), english ability(45), poverty(400), renters(108), group quarters(38), commuting, marriage(128), marital status(23) children(329), male(460), female(460), ages(1300), military service(30), industry(20), ancestry(118), ethnicity(160), disability(48), type of housing, vehicles(48), phones(15), housing price, heating fuel(11), plumbing(63), housing costs(175), rent(108), source of water(8), source of income, head of household(900), and MUCH more, for any area in the US directly from your desktop. CensusCD can easily VIEW and PRINT this data, or EXPORT it to other packages such as: SAS, SPSS, Mapinfo, Arcview, Excel, Access, and Oracle.
CensusCD is the most comprehensive reference of the U.S. population and its housing available. CensusCD is the only product on the market with the complete 1990 US Census (3,500+ variables for 16 levels of US geography, from block group level up) on a single CD.
This data is originally sold by the US Census Bureau (data file titles STF3A, 3B, 3C & 3D) for about $3,000 on 67 CD-ROM's. Some data publishers have compressed the information down to 5 discs for about $500 with only basic tools. Others sell smaller subsets of the data with only limited geographic or demographic content. GeoLytics, the makers of CensusCD, have developed advanced compression technologies that make the convenience of accessing all of this data from one CD, possible. CensusCD finally makes access to the ENTIRE US Census PRACTICAL, AFFORDABLE, and FAST!
CensusCD's easy to use Windows interface lets users:
- Make CUSTOM DEMOGRAPHIC (from 3,500+ choices) selections
- Make CUSTOM GEOGRAPHIC (from 375,000+ areas) selections
- Select from 16 AREA TYPES: Block Group, Tract, County, State, Division, Region, Nation, ZIP-code, Place, MCD, MSA, 104th Congress, Urban, PMSA, ANRC, and Native American
- Create RADIUS area in miles around an address, zip code, place name, or latitude/longitude coordinates, for demographic reporting
- SEARCH for DEMOGRAPHIC variables by specific terms (e.g. "Education") with the counts search
- SEARCH for GEOGRAPHIC names in the entire US or a single state (e.g. all occurrences of Springfield)
- Input an address (ZIP & Street) to get neighborhood demographics (GEOCODING to the block group)
- Easily BREAK DOWN reporting areas into smaller units (e.g. all counties in California, all ZIPs in US)
- SORT (rank order) results in ascending or descending order
- Always know your next step through AUTOMATIC ADVICE which appear at the bottom of the program's window
- Access over 25,000 lines of clear & extensive HELP including the complete set of STF-3 documentation
- Produce reports in many formats, including ASCII and Dbase
- Include latitude/longitude CENTROIDS, for every geographic area (for use by mapping packages)
- VIEW, search, COPY into clipboard, or PRINT reports
- Run reports in MULTITASKING mode
CensusCD requires an IBM compatible PC with at least a 386 processor, Windows (3.1x, 95, NT), a CD-ROM drive, 8 Mb of memory, and at least 2 Mb of free hard disk space.
Visit our web site at http://www.censuscd.com for more information about CensusCD. Link to http://www.censuscd.com/features_f.htm for more features information, including screen shots and sample reports. At the same link you can also download a complete description of the 3500+ demographics available in CensusCD. Link to http://www.censuscd.com/tour_f_1.htm and take a GUIDED TOUR of CensusCD.
CensusCD provides a superior interface, unmatched convenience, extensive on-line help, geocoding, free technical support, and A 30 DAY MONEY BACK GUARANTEE, for only $199.95 (shipping and handling included).
For more information, or to order by credit card, call 1-800-577-6717, or 1-732-651-2000. Purchase orders can be faxed to 1-732-651-2721, or mailed to PO Box 10, East Brunswick, NJ 08816 USA. Questions can be sent to info-at-censuscd.com .
Craig Cornelius CEO, GeoLytics, Inc.
GeoLytics Main Office P.O Box 10 East Brunswick, NJ 08816 USA Phone : 800-577-6717 (for orders and information) 732-651-2000 (technical support) Fax : 732-651-2721 (for PO's) E-mail : info-at-censuscd.com (for information) Web : http://www.censuscd.com
PS. we spend thousands of hours surfing the web for e-mails of potential US Census data users, or data providers. we will not buy, rent, or sell e-mail addresses, or intentionally send to list serves or news groups. this message will only be sent once (by us) to an address. we apologize for any mistakes, or for sending to those against UCE
For reticles, try McCrone Accessories, 800-622-8122. For counters, any large scientific supplier, e.g., VWR.
No commercial interest... My opinions... Leonard Corwin Fort Dodge Animal Health (Analytical Research) Princeton, NJ 08543-0400 corwinl-at-pt.cyanamid.com
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
1) Does anyone know where to get special order reticles and graticles? My microscope supplier does not have what I need.
2) I also need to know where to buy a counter (I have no idea what the correct name is!) that allows you to keep track of the number of intersections you count in a field placement. These are small mechanical devices with a button that increases the counter.
Thanks in advance,
Robin Griffin UAB Materials and Mechanical Engineering
Message-Id: {199711242208.RAA15237-at-emlab.cb.uga.edu} Comments: Authenticated sender is {farmer-at-emlab.cb.uga.edu}
MICROSCOPIST. The University of Georgia invites applications for the position of Laboratory Coordinator of the Center for Ultrastructural Research. As this facility serves both the physical and biological sciences, familiarity with both disciplines is highly preferred. Primary responsibilities include assisting faculty and students in the use of the facility for teaching and research, and overseeing the upkeep and use of TEM, SEM, Confocal and specimen preparation facilities, including the analytical SEM (EDX and EBSP). Applicants should have demonstrated abilities in the field of microscopy with expertise in SEM, confocal, TEM, fluorescence microscopy and related techniques. This position is a full-time, 12-month appointment with full benefits. An M.S. in the biological or physical sciences and a minimum of three years experience is desired. Interested applicants should submit a letter of interest, a curriculum vitae, and names of three professional references to: Mark Farmer, Center for Ultrastructural Research, 151 Barrow Hall, University of Georgia, Athens, GA 30602. email: kundell-at-emlab.cb.uga.edu Applications received before December 31, 1997 will be given full consideration. The University of Georgia is an Equal Opportunity Employer.
Mix with 500 g sugar + water 800 mL + alcohol about 800 mL + 1 mL pepermint oil gives a nice drop of liquer after aging - but perhaps could be a health hazard.
We are examining inclusions in polished aluminium metal with SEM/EDX, and besides a small amount of naturally common oxides we are also seeing 5 to 15 uM diameter inclusions of SiC (... positively ID'd with u-probe ...). I am now re-polishing with 30uM diamond having skipped the grit stages and when I get to 6uM diamond I start seeing these inclusions. Can anyone imagine a possible source for these???
TIA and cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
I have a series of related questions that probably have no easy answers, but I'd like some input none-the-less.
Given that high pressure freezing (HPF) and freeze substitution (FS) have been shown to provide superior fixation of biological structures in a number of studies, of what value are other fixation techniques to modern scientific inquiry? Should studies using conventional chemical fixation (CCF), microwave fixation, propane jet freezing, plunge freezing, or any preparation technique other than HPF/FS be pursued? Is there any concensus in the scientific community on which systems, organelles, or structures can be adequately fixed with CCF and which require HPF/FS? If so, what criteria do we use in deciding whether or not a particular fixation protocol is "good enough"? I have read Crang and Klomparens (Artifacts in Biological EM) but have not been able to find answers to such questions.
This issue interests me from more than a mere philosophical perspective. I recently had a paper rejected solely because I used conventional chemical fixation and not HPF/FS (although both reviewers thought that the subject matter was well worthy of investigation, and one recommended acceptance). My access to a high pressure freezer is both limited and inconvenient so conducting HPF studies is no trivial matter (and I suspect this is true for most of the other biological microscopists on this list).
What's a scientist to do?
Bob
Dr. Robert R. Wise Department of Biology and Microbiology University of Wisconsin-Oshkosh Oshkosh, WI 54901
(920) 424-3404 tel (920) 424-1101 fax wise-at-uwosh.edu www.uwosh.edu/departments/biology/wise/wise.html
Please post the following positions on the MSA's Listserver. Thank you. _____________________________________________________________
TITLE: Scientist, Electron Optics, FEI Company, Hillsboro, Oregon DEPT: Components R&D REPORTS TO: Columns R&D Director
1) Job Summary:
Primarily works on projects requiring knowledge of beam technology and electron optics calculation capability. This is an intermediate level non-supervisory engineering/scientist position. It may involve some project leadership, but does not include management or supervision of a permanent or established organizational unit.
2) Position Objectives And Major Responsibilities:
a) Perform electron optical calculations to support design of electron and ion focusing columns, primarily using point sources (LMIS, Schottky emitters).
b) Perform electron optical calculations to support the design of high speed beam blanking, deflection and mass filters associated with such columns.
c) Perform and/or supervise experimental measurements of the performance of electron and ion columns. Determine if the measurements support the theoretical calculations.
d) Develop new experimental techniques to prove column performance.
3) SECONDARY RESPONSIBILITIES:
a) Maintain and upgrade electron optical calculation capability
b) Support marketing and sales with calculations for present products and future possible products.
c) Contribute to group understanding of HV, vacuum, mechanical, materials and particle beam technologies by interaction with group members and attendance at professional conferences and short courses. Distribute knowledge through in-house seminars.
d) Support customers with calculations for various methods of operating columns.
4) Minimum Education And Experience:
a) Advanced degree in physics or engineering (MS or PhD).
b) Familiarity with electron and/or ion focusing column design
c) Familiarity with experimental techniques associated with electron and/or ion beam characterization.
d) Experience with numerical methods for electronmagnetic field calculations.
e) Due to possibility of travel, must be able to obtain a passport.
5) Additional Desirable Qualifications:
a) Familiar with high brightness field emission electron and ion sources.
b) Familiar with UHV design.
c) Familiar with high voltage breakdown phenomena.
d) Familiar with surface physics and analysis techniques.
Interested candidates should either: Email: to Lisa Olivia at lolivia-at-feico.com FAX: to Lisa Olivia at 503-640-7509 Mail: to Lisa Olivia at FEI Company, 7451 N.W. Evergreen Parkway, Hillsboro, OR 97124 ______________________________________________________________
FEI Company, Hillsboro, Oregon
Position Title: Electrical Engineering Manager
Job Duties/Responsibilities: (Functional Manager) Establish written electrical engineering standards and procedures which are consistent with those used throughout FEI and which can meet ISO standards. Support New Product Development teams by providing leadership, timely planning, schedule updates, design, documentation, and support to the development process. Train EE personnel in these standards and procedures. Staff the EE group to meet our commitments, coach and support the EE group. Provide aftercare to existing products. Represent EE group in cross department meetings. (Senior Electrical Engineer) Interact with marketing and R&D personnel to define requirements for new electronics and write specifications. Plan, lead and track the development project. Design the new electronics, supervise subcontractors, oversee the prototyping and testing and creation of all necessary documentation for production and service. Train production and service personnel in the new electronics and provide aftercare for the electronics. Work on concurrent engineering teams as part of the development process.
Job Qualifications/Requirements (minimum): BSEE, 5 years experience in EE functional management, 7 years electrical design engineering experience specializing in low noise, high stability and high voltage analog electronics, verifiable record of work in a cross-functional, concurrent engineering environment, experience with manufacturing processes, proven track record for developing electronics on schedule and with quality, willing to travel 10% of the time.
Other Qualifications (desirable): Electrical CAD experience preferred
Interested candidates should either: Email: to Lisa Olivia at lolivia-at-feico.com FAX: to Lisa Olivia at 503-640-7509 Mail: to Lisa Olivia at FEI Company, 7451 N.W. Evergreen Parkway, Hillsboro, OR 97124
I just filled out an electronic subscription form for membership to listserver, but about 10 minutes ago sent an e-mail with two position openings my company has. I sent the email with the jobs before I subscribed. I want to make sure that since I did this backwards that my job postings are still going to go out on Listserver. They came from lolivia-at-feico.com. could you please confirm it is all set or let me know if I have to resend? Thank you.
Does anyone know of a good source of skin tissue (epitheal cell) samples that would be suitable for in situ molecular imaging of transdermic medication, application of creams and ointments, etc?
George
____________________________________________________________________ ____________________________________________________________________ George Sibbald, President Molecular Imaging Corporation, Biological AFM Contract Lab 9830A South 51st Street, Suite A124 Phoenix, AZ 85044, USA Phone(602)753-4311, Fax(602)753-4312 http://www.molec.com/
____________________________________________________________________ ____________________________________________________________________ George Sibbald, President Molecular Imaging Corporation; Technology leader "in situ" SPM 9830A South 51st Street, Suite A124 Phoenix, AZ 85044, USA Phone(602)753-4311, Fax(602)753-4312 http://www.molec.com/
} } I would be interested in hearing opinions about the features } } that microscopists consider essential on a new purchase of a confocal } } microscope for biological work. } } } } Which scopes do you consider worthy, user friendly, cost efficient } } and the best quality optically ? Are there any features or scopes } } that have proved, in use, not to perform up to expectations ? } } I would welcome input from users and vendors. } } Thanks, } } Linda M. Fox
} Our lab is also thinking of purchasing a confocal. I'd love to be } included in this discussion if it goes to private mail. } Paul Tiseo
This is a serious question. Like Bob, we had some difficulty getting a paper accepted recently because we had used conventional chemical fixation, even though in a previous paper on a very similar topic (immuno-EM of cytoskeleton) we showed that the density of antibody label (5 nm gold) was very similar in chem fixed and rapid (plunge) frozen/freeze sub (RF/FS) material. The only difference between chem fix and RF/FS was in the density of background label, which was lower in RF/FS. At the time we did the more recent work, we did not have access to RF/FS apparatus, and felt that the considerable extra time required to get the almost identical results on RF/FS material was not warranted.
I certainly do not deny that RF/FS gives superior preservation, but with some larger tissues when you are looking at structures/cells deep in the tissue and do not want to induce artefacts due to cutting the tissue, chemical and other non-freezing methods are the only way to go. We deal with plant tissues, which have severe freezing damage at the EM level below about 20 or so =B5m in plunge frozen material, and below maybe 40 or so =B5m= in high pressure frozen (HPF) material (can get much deeper preservation in certain tissues). THe quality of preservation is extremely variable at the EM level, and you have to section through lots of material, even after processing using automated apparatus, in order to get cells with minimal ice damage. I guess what we need to know is what are the consistent artefacts induced by chemical or other non-freezing fixation, and can we take these into account and still get useful information from this work.
Now for light microscope work, esp. immunofluorescence (at least of plant tissues), RF/FS is great, giving consistently superior preservation and antigenicity than after chemical fixation, even for large tissues.
Re. HPF - there are some consistent, worrying artefacts. In HPF/FS plant material, the ER always seems to look "blown up" like little sausages or balloons, and the ER membrane is often almost impossible to see. Is this effect seen in animal tissues processed in this way? Other work has shown that a tubular membrane network, seen after RF/FS (plunging or slamming), was broken into a series of vesicles by HPF/FS. Unsupported membranes seem to be a problem.
So I guess you have to argue your case each time. We are currently spending some time doing a useful if somewhat tedious study comparing various fixation methods with RF/FS and HPF/FS for just this reason. It is important because the fine ultrastructure of some cell structures is still contentious, and we need to know if differences reported by different groups are due to various fixation artefacts or to differences in species used, tissue type, etc.
I would also be interested to hear of others' experiences and opinions.
regards,
Rosemary
Rosemary White Department of Biological Sciences Monash University, Melbourne, Victoria 3168, Australia phone 61-3-9905 5670 fax 61-3-9905 5613 email r.g.white-at-sci.monash.edu.au
} } Dear biological ultrastructuralists, } } I have a series of related questions that probably have no easy answers, } but I'd like some input none-the-less. } } Given that high pressure freezing (HPF) and freeze substitution (FS) have } been shown to provide superior fixation of biological structures in a } number of studies, of what value are other fixation techniques to modern } scientific inquiry? Should studies using conventional chemical fixation } (CCF), microwave fixation, propane jet freezing, plunge freezing, or any } preparation technique other than HPF/FS be pursued? Is there any concensus } in the scientific community on which systems, organelles, or structures can } be adequately fixed with CCF and which require HPF/FS? If so, what } criteria do we use in deciding whether or not a particular fixation } protocol is "good enough"? I have read Crang and Klomparens (Artifacts in } Biological EM) but have not been able to find answers to such questions. } } This issue interests me from more than a mere philosophical perspective. I } recently had a paper rejected solely because I used conventional chemical } fixation and not HPF/FS (although both reviewers thought that the subject } matter was well worthy of investigation, and one recommended acceptance). } My access to a high pressure freezer is both limited and inconvenient so } conducting HPF studies is no trivial matter (and I suspect this is true for } most of the other biological microscopists on this list). } } What's a scientist to do? } } Bob } } Dr. Robert R. Wise } Department of Biology and Microbiology } University of Wisconsin-Oshkosh } Oshkosh, WI 54901 } } (920) 424-3404 tel } (920) 424-1101 fax } wise-at-uwosh.edu } www.uwosh.edu/departments/biology/wise/wise.html
Hello Robert,
I think - although I did not yet have the opportunity to experience it - high pressure freezing (HPF) followed by freeze substitution (SF) is a technique that provides most excellent results. However, it is also one of the most highly sophisticated techniques - not to mention the costs of high pressure freezer (in Germany about 250 000.- DM) and substitution unit (about 50 000.- DM), roughly estimated.
As with every technique, it has its limitations. And reminding what I have seen in the literature, the most severe restriction is that you need very small sample sizes (see e.g. Hohenberg et al. J. Microscopy Vol.175: 34-43, 1994). Largest samples sizes are 400 micron in thickness reported to be well preserved (e.g. Studer et al., Scanning Microsc. Vol.3: 253-69, 1989).
So what to doe if the specimen of interest is larger than 400 micron ?
If you cut it down to pieces of that small size, you have to face the problem that this induces alterations in ultrastructure that will increase severely with time until fixation. From a microanalytical point of view, Thomas von Zglinicki (J. Microsc. Vol.141:79-90, 1986) has shown that stopping blood flow for more than 10 s (!!) results in ion shifts between cells and extracellular space in rat heart and liver. And there is also a stereological paper that I cannot find at the moment which describes changes within a short period of time after death of the experimental animal.
If you use cryosnappers or other devices to cryofix small samples in situ with the blood still flowing, you have to face the problem that you may only reach the cortex of your object of interest - but what to do if your are interested in the ultrastructure of the medulla ? Further, you will be able to collect only a limited number of samples - is this collection representative of your object or is it a rather biased collection (see sterelogy literature - e.g. Exp. Physiol. Vol.76:639-65, 1991; J. Anatomy Vol.188,1-15, 1996) ? If you are looking for differences between groups of animals (developmental stages, treatments etc.) your comparison may rely on excellently preserved ultrastructure, but you will not be able to exclude that the differences seen between groups are due to the biasedness of your sample collection.
So I conclude that for most purposes conventional chemical fixation still has its merits, and is frequently the only way to obtain any ultrastructural data.
I will not accept that HPF/SF has to be performed in any study of biological material.
Heinz
**************************************************************************** Dr. Heinz Fehrenbach Institute of Pathology University Clinics "Carl Gustav Carus" Technical University of Dresden
Does anyone have a good reference about how to do a two-dimensional Penrose Tiling as in Decagonal Quasicrystals?
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University Evanston, IL 60208-3108 tel: (847) 491-3996 fax: (847) 491-7820 email: l-marks-at-nwu.edu http: //www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
I am buying a Carl Zeiss transmission trinocular microscope with a video camera. I asked the seller to include an adaptor ring, just in the case we can gather some more money to buy a photo camera in the future. The seller asks me to specify a trademark, because the ring is specific for each camera. I am completely ignorant about this facts, so I am asking for your advice.
In short: what do you consider the best camera relating cost/performance, so as to order the corresponding adaptor? We intend to photograph erythrocytes submitted to strain, and also crystalline samples.
Thank you very much in advance for any help you may give to me.
Ana _________________________________________________________________
Ana Maria Gennaro Grupo de Fisica- INTEC
Home address: Sarmiento 6602 Work address: INTEC - Guemes 3450 3000 - Santa Fe 3000 - Santa Fe Argentina Argentina Phone: 54-42-606764 Phone: 54-42-559175/6/7 Fax: 54-42-550944
Michael Shaffer asked about possible sources of SiC contamination in polished aluminum samples.
Michael, You mentioned that originally you found SiC after using SiC papers and you positively identified these particles using microprobe. Then you stated that you switched to diamond, eliminating the SiC papers from the procedure. Did you then do microprobe analysis again? The reason I ask is that embedded diamond particles and SiC particles do look very similar in bright field LOM. In addition, diamond does polish diamond, and you may be seeing a flat face on the particles, therefore assuming they are SiC.
If you have possitively identified this second batch of particles as SiC, and there is no possibility of these particles coming from the melt, then I would have to suggest that contamination from the polishing system may be the culprit. (Time to clean the lab).
Good luck and Best regards,
Scott D. Holt BUEHLER, LTD. PO Box 1 41 Waukegan Rd. Lake Bluff, IL 60044 (847)295-6500, Ext. 4546 http://www.buehlerltd.com
Dear People, since I originated the discussion I also want to add, what is our need for high pressure freezing. It is NOT the ultrastructure alone in our case, we want to do high resolution ( {= 1 micron) resolution element mappings AND quantification. The probelm is, that this is a resolution already disturbed by ice crystals. On the other hand, if HPF is limited to 40-400 microns (one probably has to try?), it is at the limit also for things like roots - if not Arabidopsis ;-). However there will probably be much less ice damage and thus more information - if it will be possible to cut the samples frozen or if freeze substitution itself causes not to many loss (or REDISTRIBUTION) of material. A lot of problems I think, but therefore one probably should use the best method. However, who is able to pay it... Arthur
Dr. Arthur Schuessler University of Heidelberg Zellenlehre Im Neuenheimer Feld 230 D-69120 Heidelberg Germany
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I am buying a Carl Zeiss transmission trinocular microscope } with a video camera. I asked the seller to include an adaptor } ring, just in the case we can gather some more money to } buy a photo camera in the future. } The seller asks me to specify a trademark, because the ring is } specific for each camera. I am completely ignorant about this } facts, so I am asking for your advice. } } In short: what do you consider the best camera relating } cost/performance, so as to order the corresponding adaptor? } We intend to photograph } erythrocytes submitted to strain, and also crystalline samples. } } Thank you very much in advance for any help you may give to me. } } Ana } _________________________________________________________________ }
Ana, in your present circumstances (unsure what to buy, not ready to buy now, etc.) the best course is to maximize your future compatibility with minimal current expenditure (that might be made useless by the emergence of new equipment or standards). Ask your Zeiss Rep to provide two couplings: For 35 mm film photography ask for a standard "T-Mount" adapter on the microscope. For video micrography or digital photomicrography, as for a standard "C-Mount" adapter. You can later get a T-Mount adapter for virtually any 35 mm camera you buy which will mate with the one on the microscope. Similarly, most digital and video cameras come with a C-Mount thread which should mate directly to the one on the microscope. When you shop for a video or digital camera, be sure that it has a removable lens and a standard C-Mount thread so that you can mount your own lenses or the adapter to the microscope easily.
Hope this helps.
-- ********************************************************** Stephen A. Shaffer sshaffer-at-microdataware.com MicroDataware http:www.microdataware.com (Under reconstruction and temporarily out of service) Personal stuff: steve_shaffer-at-compuserve.com http://ourworld.compuserve.com/homepages/steve_shaffer/ **********************************************************
My friend need a glue for sectional TEM specimen preparation. I could not find a easy way to contact the company listed following. If you happen to know the phone Num. and/or email address of this company, please send me an email.
if you are interested in this workshop please respond directly to Xavier Llovet {xavier-at-giga.sct.ub.es} , who is *not* subscribed to the microscopy list. 1 USD = about 125 pesetas.
regards, YM.
---------- Forwarded message ----------
shAf:
Please pardon the simplicity of these suggestions, as you may have thought of them already....
1. Could the SiC be intrinsic to your specimen? There is more DRA Al alloy in the world today than people realize. "DRA" is "Directionally Reinforced Aluminum;" the usual reinforcement is SiC particles. Have you thought of electropolishing your specimen surface deeply enough to access unaltered material, then checking for SiC particles on that surface with SEM+EDXS, or EMP?
2, Are SiC particles still being introduced during grinding/polishing as a contaminant within the nap, or on the surface, of one or more of your diamond wheels?
Dave Snow
--------------------------------------------------------------------- David B. Snow Materials Analysis Group Pratt & Whitney, Materials & Mechanics Engineering East Hartford, CT 06108 860 565 7823 snowdb-at-pweh.com
========================================
ORIGINAL MESSAGE:
----------------------------------------------------------------------- - The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} Given that high pressure freezing (HPF) and freeze substitution (FS) have } been shown to provide superior fixation of biological structures in a } number of studies, of what value are other fixation techniques to modern } scientific inquiry? Should studies using conventional chemical fixation } (CCF), microwave fixation, propane jet freezing, plunge freezing, or any } preparation technique other than HPF/FS be pursued? Is there any concensus } in the scientific community on which systems, organelles, or structures can } be adequately fixed with CCF and which require HPF/FS? If so, what } criteria do we use in deciding whether or not a particular fixation } protocol is "good enough"?
} What's a scientist to do? } } Bob } } Dr. Robert R. Wise } Department of Biology and Microbiology } University of Wisconsin-Oshkosh } Oshkosh, WI 54901
Hi Bob, I've had to address this question recently while assisting a colleague with a grant proposal. It is true that the HPF/FS is a superior combination technique regarding the preservation of antigenicity and ultrastructure for many tissues both plant and animal. If only every lab had a HPF device.... But......FWIW
HPF and FS are not without their own artifacts and pitfalls. HPF is most useful for freezing easily accessed, small samples no larger than ~1mm dia due to sample holder constraints. Not every sample can fit this requirement. Conventional chemical fixation still is a viable stabilization means, depending upon what you are ultimately investigating.
My position is to always, if possible, use multiple fixation techniques on duplicate samples. If there is a question regarding possible artifact or interpretation of a non-conventional sample then one has an alternate sample or samples to query. Besides, as you mention equipment accessibility is also an issue.
I'm not absolutely convinced that the "number of studies" indicating superior fixation with HPF/FS constitues an absolute criteria to abandon all other fixation methods. If it's available by all means use it but I would want to keep conventionally fixed backup samples in reserve, just in case. The most important step is the initial treatment of the sample. In any protocol something can always go awry.
In my opinion, there are still too few comparative studies to adequately come to a consensus. My bottom line is: It depends on what I am trying to demonstrate, what resolution level I need and what means are available to me.
cheers ed
Edward J. Basgall, PhD The Pennsylvania State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618 http://www.personal.psu.edu/ejb11/ Privilege does not absolve one of ecological responsibility.
Dear Folks, I am looking for an image (electronic, hard copy etc.) of a collage of scientific instruments i.e. microscopes, NMR, mass spec., etc. If you can help please respond to: Leo Porter at Lporter-at-goodyear.com. Thanks for your kind help.
Dear Folks, I am looking for an image (electronic, hard copy etc.) of a collage of scientific instruments i.e. microscopes, NMR, mass spec., etc. If you can help please respond to: Leo Porter at Lporter-at-goodyear.com. Thanks for your kind help.
This discussion raises the question of why freeze?
Is this the bastion of SEM?
Or are many molecular biologists not aware of in situ Molecular Imaging biological samples at 37 C?
George
At 09:42 AM 11/25/97 +0100, Heinz Fehrenbach wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
____________________________________________________________________ ____________________________________________________________________ George Sibbald, President Molecular Imaging Corporation, Biological AFM Contract Lab 9830A South 51st Street, Suite A124 Phoenix, AZ 85044, USA Phone(602)753-4311, Fax(602)753-4312 http://www.molec.com/
Praxair Surface Technologies, Inc, a Fortune 500 Company and world leader in surface enhancement, has a challenging opportunity available in our Indianapolis, IN offices.
As an Instrument Analyst, you will operate a Scanning Electron Microscope with associated instrumentation like EDS and WDS and X-Ray Diffraction equipment to provide surface analysis, chemical composition and phase analysis. This position supports the function of several engineers and research scientists in a market driven Research and Development environment.
The preferred candidate will have a BS in Chemistry, Physics or Materials with 2-3 years experience in a similar environment.
We offer a competitive salary, benefits and profit sharing. For consideration, send resume including salary history to: Praxair Surface Technologies, Inc. Attn: Linda Baber, 1500 Polco Street, Indianapolis, IN 46224. Fax 317-240-2260, E-mail: lbaber-at-geof.psti.praxair.com.
We are examining inclusions in polished aluminium metal with SEM/EDX, and besides a small amount of naturally common oxides we are also seeing 5 to 15 uM diameter inclusions of SiC (... positively ID'd with u-probe ...). I am now re-polishing with 30uM diamond having skipped the grit stages and when I get to 6uM diamond I start seeing these inclusions. Can anyone imagine a possible source for these???
TIA and cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
} I am buying a Carl Zeiss transmission trinocular microscope } with a video camera. I asked the seller to include an adaptor } ring, just in the case we can gather some more money to } buy a photo camera in the future. } The seller asks me to specify a trademark, because the ring is } specific for each camera. I am completely ignorant about this } facts, so I am asking for your advice. } } In short: what do you consider the best camera relating } cost/performance, so as to order the corresponding adaptor? } We intend to photograph } erythrocytes submitted to strain, and also crystalline samples.
The question also depends on what type of camera you are considering: a standard SLR body or a dedicated auto/manual microscope camera.
Regarding ultrastructure revealed by cryopreservation vs chemical fixation:
We have also struggled with this issue in our laboratory. We have been fortunate to prepare several types of connective tissue by HPF and indeed, the ultrastructure in well frozen areas is VERY different from that following chemical fixation and also microwave processing. For example, when viewing basement membranes, we and other laboratories have noticed an immense difference in ultrastructure. The region termed lamina densa is determined to be an artifact, probably caused by the precipitation of the many concentrated molecules present in this general region which are involved in the adhesion of epithelium to CT. The lamina lucida is also a fixation artifact, arising as the epithelial cell shrinks away from the underlying CT matrix. These are examples of serious artifacts directly attributable to chemical processing which one does not see in well frozen tissue. However, interestingly, there are several structures which may be seen in chemically processed tissue are generally not visible in cryoprocessed tissue. Ironically, if we had started out using cryoprocessing, we might not know of the existence of these structures. Two fiber systems in the CT matrix come to mind. Anchoring fibrils, which "staple" the epithelium onto the underlying CT; another are matrix microfibrils, which ensheathe elastin bundles. The concentration of proteoglycans in the matrix is so dense (and well retained by HPF) that these structures are virtually invisible (masked by such a high PG concentration). These proteoglycans are extracted in most chemical fixation protocols, allowing the visualization of anchoring fibrils and microfibrils. The lack of anchoring fibrils causes a severe blistering disease, and the defect of a microfibril component called fibrillin results in Marfans syndrome.
It is currently unrealistic for all Investigators to examine tissue by HPF. It is technically difficult, very expensive and quite time consuming to find well frozen areas. However, we should all be aware of the structure revealed by cryopreservation methodology so that we might most adequately be aware of the artifacts present in the tissues we examine. ---------------------- Doug Keene Director EM Portland Shriners Hospital DRK-at-shcc.org
shAf wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } We are examining inclusions in polished aluminium metal with SEM/EDX, } and besides a small amount of naturally common oxides we are also seeing } 5 to 15 uM diameter inclusions of SiC (... positively ID'd with u-probe } ...). } I am now re-polishing with 30uM diamond having skipped the grit stages } and when I get to 6uM diamond I start seeing these inclusions. Can } anyone imagine a possible source for these??? } } TIA and cheerios, shAf } -- } {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} } Michael Shaffer, R.A. - University of Oregon Electron Probe Facility } mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu } http://darkwing.uoregon.edu/~mshaf/
I would u probe my 6u "diamond" just to be sure, especially since it "shows up" after a 6u diamond polish.
Ken Converse owner Quality Images third party SEM maintenance
Lisa Olivia wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } hello: } } I just filled out an electronic subscription form for membership to } listserver, but about 10 minutes ago sent an e-mail with two position } openings my company has. I sent the email with the jobs before I } subscribed. I want to make sure that since I did this backwards that } my job postings are still going to go out on Listserver. They came from } lolivia-at-feico.com. could you please confirm it is all set or let me } know if I have to resend? Thank you. Yup, They got out.
Ken Converse owner Quality Images third party SEM sevice
Ana Maria Gennaro wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } I am buying a Carl Zeiss transmission trinocular microscope } with a video camera. I asked the seller to include an adaptor } ring, just in the case we can gather some more money to } buy a photo camera in the future. } The seller asks me to specify a trademark, because the ring is } specific for each camera. I am completely ignorant about this } facts, so I am asking for your advice. } } In short: what do you consider the best camera relating } cost/performance, so as to order the corresponding adaptor? } We intend to photograph } erythrocytes submitted to strain, and also crystalline samples. } } Thank you very much in advance for any help you may give to me. } } Ana } _________________________________________________________________ } } Ana Maria Gennaro Grupo de Fisica- INTEC } } Home address: Sarmiento 6602 Work address: INTEC - Guemes 3450 } 3000 - Santa Fe 3000 - Santa Fe } Argentina Argentina } Phone: 54-42-606764 Phone: 54-42-559175/6/7 } Fax: 54-42-550944 } } _________________________________________________________________ } } Dear Ana,
The type of camera you purchase depends on a number of parameters. First, you need to decide if you want to rely on regular photography or to investigate videomicroscopy. For
P.O. BOX 27777 Raleigh, North Carolina, 27611 USA.
International customer service specialist: Marsha GAYHARDT (09 95).
************************************************************************ } } My friend need a glue for sectional TEM specimen } preparation. I could not find a easy way to contact } the company listed following. If you happen to know } the phone Num. and/or email address of this company, } please send me an email. } } Thanks } } *************************************************** } M:M M-LINE accessories } measurements group, INC., RALEIGH, N.C. } M-BOND 610 ADHESIVE KIT CONTENTS:4 Unmixed, } Premeasured Units } ***************************************************
It has been a few years since I have been involved with temperature control stages on optical microscopes. Last I remember there were issues such as surface area temperature mapping(variations) functions, isolation, target range overshooting, steady state vs. drift functions, heating and cooling techniques, depth of focus and other optical issues, maximum and minimum feasible temperatures, heating and cooling techniques, etc. Can anyone refresh my memory , bring me up to date ,and refer me to journal and review articles on the subject. I would like references to current manufacturers. Also I would be interested in personal experiences with specific current equipment manufacturers of both microscopes and control stages for this use (preferably any potentially negative feedback opinions sent only to my personal email address below, so no manufacturer is embarrassed or damaged by personal opinions in this forum/possibly an obvious point of controversy). Finally does anyone have any personal experience with modifications to existing designs or references to in house construction of these devices. I also might be prospect for use equipment of this nature.
JD EMAIL1: wa5ekh-at-juno.com please cc to EMAIL2: wa5ekh-at-cyberramp.net
{html} {head} {/head} {BODY bgcolor=3D"#FFFFFF"} {p} {font size=3D2 = color=3D"#000000" face=3D"Arial"} It has been a few years since I have = been involved with temperature control stages on optical microscopes. = Last I remember there were issues such as surface area temperature = mapping(variations) functions, isolation, target range overshooting, = steady state vs. drift functions, heating and cooling techniques, depth = of focus and other optical issues, maximum and minimum feasible = temperatures, heating and cooling techniques, etc. Can anyone refresh my = memory , bring me up to date ,and refer me to journal and review = articles on the subject. {br} I would like references to current = manufacturers. Also I would be interested in personal experiences with = specific current equipment manufacturers of both microscopes and control = stages for this use (preferably any potentially negative feedback = opinions sent only to my personal email address below, so no = manufacturer is embarrassed or damaged by personal opinions in this = forum/possibly an obvious point of controversy). {br} Finally does = anyone have any personal experience with modifications to existing = designs or references to in house construction of these devices. = {br} I also might be prospect for use equipment of this = nature. {br} {br} JD {br} �= 9; EMAIL1: {font = color=3D"#0000FF"} {u} wa5ekh-at-juno.com {/u} {font color=3D"#000000"} = {br} please cc to = EMAIL2: {font = color=3D"#0000FF"} {u} wa5ekh-at-cyberramp.net {/u} {font color=3D"#000000"} = {br} {br} {font size=3D2 color=3D"#008080"} {br} {/p} {/font} {/font} {/font} {/font} {/font} {/font} {/body} {/html} ------=_NextPart_000_01BCFA1F.3793E220--
We would appreciate any info about possible source in Europe of buying regularly all ranges of sandpaper made by German company named HERMES. The best would be manufacturers direct contact. ...or other manufacturer of round shaped (230 mm, 300mm) selfadhesive, waterproof sandpaper.
kind regards
Krzysztof M. Herman LabSoft S.c. 05-500 Piaseczno, 21 Kosciuszki Str., Poland tel/fax: (48 22) 7502024, 7502028, 7570671, mobile: (48 90)213438 fax only: (48 22) 483787, Email: labsoft-at-labsoft.com.pl
I am intending to examine cells treated with an acridine-linked DNA oligonucleotide via UV microscopy. If possible I would like any information you may have regarding the filter sets I will require. Any help would be gratefully received. _______________________________________________ Andrew GRIGG Genito-Urinary Medicine Imperial College School of Medicine (St.Mary's) Norfolk Place, LONDON W2 1PG Email: a.grigg-at-ic.ac.uk _______________________________________________
My problem: a Li-Co-Oxide deposited on Al was submitted as an "unknown" to a local analytical lab. Outfitted with an SEM and an EDS capable of light element characterization, the lab reported back an analysis without any Li.
Examination of the printout of the spectrum shows a peak centered at very low keV (Li k-alpha is at 0.05391 keV). This low-energy peak apparently was not "characterized" during the peak analysis routine. As plotted (energy scale is from 0-10 keV), the peak is truncated on its low-energy side, thus only about 3/4 of the peak is displayed.
My question has two parts:
1. is Li amenable to analysis by EDS?, and, if Li is amenable to such,
2. is the lack of Li in the analysis due to the inability of the peaking routine to 'identify' (quantify) a peak which is not fully displayed? (to fully display the peak, the energy scale would have to begin at approx. -0.5 keV).
Winton Cornell
P.S. the analysis shows 'good' results for O, low Al (as hoped, because the substrate was nicely coated), significant carbon (because of the carbon coating), no Li, and 'low' Co........all in all, not a 'good', or representative analysis as far as we're concerned
Dr. Winton Cornell Senior Research Associate & Supervisor, Microanalysis Laboratory Department of Geosciences The University of Tulsa 600 South College Tulsa, OK 74104-3189
I am analyzing calcite shells in the TEM. I am having difficulty gluing the calcite samples onto the grid. I have been using colloidal silver liquid, but it either dries before I can get the calcite in place or I get too much liquid and the grid "drowns". Is this just a matter of practice, or is there another gluing material that I can try that is more forgiving ?
Thanks.
Nancy Buening
Nancy Buening E-mail: buening-at-geology.ucdavis.edu Department of Geology Phone: (530) 752-0350 University of California Fax: 530 752-0951 Davis, CA 95616
Hi there we are looking for a very small objective aperture for our JEOL 2000FX. Since these apertures come in strips there is a limited selection and the "standard" set comes with a 20micorn aperture as the smallest. We can get a 10micron from JEOL, does anyone know of a smaller one say 5microns? I have not called all of the Microscope Spares and Equipment suppliers yet, I thought I would solicit commnets for the community. Many thanks. Reply by email and I will summarize to the list if there is sufficient interest and response.
Cheers
Jfm.
John Mansfield North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (313) 936-3352 FAX (313) 936-3352 Cellular Phone: (313) 715-2510 (Leaving a phone message at 936-3352 is preferable to 715-2510) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html
It's not possible to analyze for Li in a normal bulk sample. To generate a K-alpha x-ray, you need an electron transition from the 2p to the 1s state. If you look at the periodic table, Li has only a single 2s electron. You can't have a 2s to 1s transition because the x-ray has 1 h/(2*pi) of angular momentum and this transition would violate conservation of angular momentum.
The peak that you see at that low energy is low energy noise.
-Scott
Scott D. Walck PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of S.D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
My problem: a Li-Co-Oxide deposited on Al was submitted as an "unknown" to a local analytical lab. Outfitted with an SEM and an EDS capable of light element characterization, the lab reported back an analysis without any Li.
Examination of the printout of the spectrum shows a peak centered at very low keV (Li k-alpha is at 0.05391 keV). This low-energy peak apparently was not "characterized" during the peak analysis routine. As plotted (energy scale is from 0-10 keV), the peak is truncated on its low-energy side, thus only about 3/4 of the peak is displayed.
My question has two parts:
1. is Li amenable to analysis by EDS?, and, if Li is amenable to such,
2. is the lack of Li in the analysis due to the inability of the peaking routine to 'identify' (quantify) a peak which is not fully displayed? (to fully display the peak, the energy scale would have to begin at approx. -0.5 keV).
Winton Cornell
P.S. the analysis shows 'good' results for O, low Al (as hoped, because the substrate was nicely coated), significant carbon (because of the carbon coating), no Li, and 'low' Co........all in all, not a 'good', or representative analysis as far as we're concerned
Dr. Winton Cornell Senior Research Associate & Supervisor, Microanalysis Laboratory Department of Geosciences The University of Tulsa 600 South College Tulsa, OK 74104-3189
Our Link Isis system has a "zero strobe" peak which occurs at 0.0 kV. Since it is not indicative of anything in the sample, we usually cut it off when we plot our spectra. Our spectra pretty well tail off at 0.1 kV and the zero strobe runs up to 0.1 kV under our conditions. So no, Li could not be handled with our detector, as good as it is. There seem to be some experimental detectors on their way that could do Li, but I doubt that your service lab was running one of them.
At 11:45 AM 11/26/97 -0600, you wrote: } Listers: } } My problem: a Li-Co-Oxide deposited on Al was submitted as an "unknown" to } a local analytical lab. Outfitted with an SEM and an EDS capable of light } element characterization, the lab reported back an analysis without any Li. } } } Examination of the printout of the spectrum shows a peak centered at very } low keV (Li k-alpha is at 0.05391 keV). This low-energy peak apparently was } not "characterized" during the peak analysis routine. As plotted (energy } scale is from 0-10 keV), the peak is truncated on its low-energy side, thus } only about 3/4 of the peak is displayed. } } My question has two parts: } } 1. is Li amenable to analysis by EDS?, and, if Li is amenable to such, } } 2. is the lack of Li in the analysis due to the inability of the peaking } routine to 'identify' (quantify) a peak which is not fully displayed? (to } fully display the peak, the energy scale would have to begin at approx. } -0.5 keV). } } Winton Cornell } } P.S. the analysis shows 'good' results for O, low Al (as hoped, because the } substrate was nicely coated), significant carbon (because of the carbon } coating), no Li, and 'low' Co........all in all, not a 'good', or } representative analysis as far as we're concerned } } } Dr. Winton Cornell } Senior Research Associate & Supervisor, Microanalysis Laboratory } Department of Geosciences } The University of Tulsa } 600 South College } Tulsa, OK 74104-3189 } } phone: 918-631-3248 } fax: 918-631-2091 } e-mail: wcornell-at-centum.utulsa.edu } } } ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-8216
} My problem: a Li-Co-Oxide deposited on Al was submitted as an "unknown" to } a local analytical lab. Outfitted with an SEM and an EDS capable of light } element characterization, the lab reported back an analysis without any Li. } } Examination of the printout of the spectrum shows a peak centered at very } low keV (Li k-alpha is at 0.05391 keV). This low-energy peak apparently was } not "characterized" during the peak analysis routine. As plotted (energy } scale is from 0-10 keV), the peak is truncated on its low-energy side, thus } only about 3/4 of the peak is displayed. } } My question has two parts: } } 1. is Li amenable to analysis by EDS?, and, if Li is amenable to such, } 54 eV photons may not be analysable due to the inability of a Si(Li) detector to produce enough electron-hole pairs (EHP) to produce a measurable pulse. At 3 eV energy loss per EHP there would be only 18+-4 EHP produced if all the Li k-alpha photon energy was lost in the active volume. Recombination would reduce this number somewhat, and the re- maining EHP must be significantly more than the dark current of the detector. This best-case (!) scenario assumes that there is no dead layer in the detector and that the photon is produced at the surface of the specimen (no absorption). EELS is a better way to analyse for Li, but you would need a thin specimen so that plasmon losses don't obli- terate the signal (Al has ~15 eV plasmons, so the 4-plasmon peak would overlap the signal, which is the k-ionization energy--slightly } 54 eV).
} 2. is the lack of Li in the analysis due to the inability of the peaking } routine to 'identify' (quantify) a peak which is not fully displayed? (to } fully display the peak, the energy scale would have to begin at approx. } -0.5 keV). } My guess is that at 54 eV there is too much noise in a Si(Li) detector, even at LN2 temperature, and in the electronics to get a real peak. The FWHM of the peak would be ~20 eV (I don't have the # of SD's in a FWHM memorized or nearby), so all the noise within this region must be compared to the expected signal. The experts in the EDX field would know for sure. Comments from WDS experts would be good too. Yours, Bill Tivol
} ... } At 11:45 AM 11/26/97 -0600, Dr. Winton Cornell wrote: } } Listers: } } } } My problem: a Li-Co-Oxide deposited on Al was submitted as an "unknown" to } } a local analytical lab. Outfitted with an SEM and an EDS capable of light } } element characterization, the lab reported back an analysis without any Li. } } } } } } Examination of the printout of the spectrum shows a peak centered at very } } low keV (Li k-alpha is at 0.05391 keV). This low-energy peak apparently was } } not "characterized" during the peak analysis routine. ... } } My question has two parts: } } } } ... }
Our Oxford EDX synthetically generates a "zero" peak for the purpose of properly offsetting the energy axis. The computer will*not* label this peak. So as not to cause confusion we generally label it "00" ...
cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
We have an LKB 2178 Knifemaker II that needs servicing - it takes a lot of pressure before it will score the glass and then chips the glass edge rather than a clean fracture. Does anyone know of someone who services these in the Montreal, Canada area?
Secondly, we need parts for a Reichert FC4 unit - specifically the upper part of the cryo knife holder and the specimen (not chuck) set screw and allen key. Again, does anyone know who sells these to the Montreal area?
Thanks in advance,
Pat Hales McGill University Dept. of Anatomy & Cell Biology hales-at-hippo.medcor.mcgill.ca
Winton Cornell wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Listers: } } My problem: a Li-Co-Oxide deposited on Al was submitted as an "unknown" to } a local analytical lab. Outfitted with an SEM and an EDS capable of light } element characterization, the lab reported back an analysis without any Li. } } Examination of the printout of the spectrum shows a peak centered at very } low keV (Li k-alpha is at 0.05391 keV). This low-energy peak apparently was } not "characterized" during the peak analysis routine. As plotted (energy } scale is from 0-10 keV), the peak is truncated on its low-energy side, thus } only about 3/4 of the peak is displayed. } } My question has two parts: } } 1. is Li amenable to analysis by EDS?, and, if Li is amenable to such, } } 2. is the lack of Li in the analysis due to the inability of the peaking } routine to 'identify' (quantify) a peak which is not fully displayed? (to } fully display the peak, the energy scale would have to begin at approx. } -0.5 keV). } } Winton Cornell } } P.S. the analysis shows 'good' results for O, low Al (as hoped, because the } substrate was nicely coated), significant carbon (because of the carbon } coating), no Li, and 'low' Co........all in all, not a 'good', or } representative analysis as far as we're concerned } } Dr. Winton Cornell } Senior Research Associate & Supervisor, Microanalysis Laboratory } Department of Geosciences } The University of Tulsa } 600 South College } Tulsa, OK 74104-3189 } } phone: 918-631-3248 } fax: 918-631-2091 } e-mail: wcornell-at-centum.utulsa.edu Li is not only below the detection level of the latest light element EDS detectors but is also below the detection limits for WDS detectors. With the right crystals they will go to Be, but no lower. The "peak" you were looking at was unsupressed amplifier noise. You will need to look at something besides x-rays if you wish to look for Li. Ken Converse owner Quality Images third party SEM service
John Mansfield wrote: ================================================= .......we are looking for a very small objective aperture for our JEOL 2000FX. Since these apertures come in strips there is a limited selection and the "standard" set comes with a 20micorn aperture as the smallest. We can get a 10micron from JEOL, does anyone know of a smaller one say 5microns ? ================================================= There are some production issues that make it difficult, if not impossible, to produce the smaller size holes on a strip. Is there any way to convert such strip aperture holders to "take" disc apertures? We have several persons wanting to use our 1 and 2 um hole (no, that is not a typo) gold foil thin film apertures but can't since their scopes take only the strips.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
} Hi there we are looking for a very small objective aperture for our JEOL } 2000FX. Since these apertures come in strips there is a limited selection } and the "standard" set comes with a 20micorn aperture as the smallest. We } can get a 10micron from JEOL, does anyone know of a smaller one say } 5microns? } I have not called all of the Microscope Spares and Equipment suppliers yet, } I thought I would solicit commnets for the community. Many thanks. } Reply by email and I will summarize to the list if there is sufficient } interest and response. } } Cheers } } Jfm. } } } John Mansfield
Hi John,
Agar certainly sell strips for the 2000FX down to 5 micron. I would expect SPI to sell them as well and the others.
The problem with these very small apertures is they get dirty very quickly, even on microscopes with 'clean' vacuum systems, since they sit very close to the specimen. For objective use, thin film gold apertures are better than solid apertures, as they 'self clean' and last a reasonable amount of time. I don't think these are available in strip form, only as singles for aperture rods like Philips. Of course, they are not much good in the condenser, as it is too easy to punch holes in them with the beam, although I can't think of a reason to go this small at the condenser.
Regards,
-- Larry Stoter 17, Rocks Park Road, Uckfield, E. Sussex, TN22 2AT, UK email: LPS-at-teknesis.demon.co.uk Phone/Fax: +44 (0)1825 767967
} We have an LKB 2178 Knifemaker II that needs servicing - it takes a lot of } pressure before it will score the glass and then chips the glass edge rather } than a clean fracture. Does anyone know of someone who services these in the } Montreal, Canada area? ----------cut---------- Dear Pat
You should not have to apply pressure at all, this is what's causing the edges to chip. A small adjustment of the scoring mechanism (check that the wheel is ok) should do the trick. Also make sure that the block does not slip during breaking e.g. don't put oil on the block holder!!! These knife makers outlive most microscopists.
Sincerely +----------------------------------------------------------------- |Dr Stephan Helfer, SSO |Senior Mycologist - MSc Course Director | |Royal Botanic Garden Edinburgh, Inverleith Row, EDINBURGH EH3 5LR, |Scotland UK | |http://www.rbge.org.uk | |phone: +44 (0)131 552 7171 ext 280 | or +44 (0)131 459 0446-280 (direct digital VoiceMail line) |fax: +44 (0)131 552 0382 +------------------------------------------------------------------
I am interested in your opinion about electrolyhtic thinning apparatues and their performance. I am planning to by a jet polishing machine to be used for making thin foils from 3mm disks. The material I am interested in is High strength Aluminium alloys, steels, stainless steels.
Best wishes=20 Sten Johansson
Link=F6ping University Department of Mechanical Engineering
Does anyone have a good basic reference for processing bone for immuno- cytochemistry? Either a paper, even an author perhaps a Web site or personal experience. It would be highly appreciated.
Thank you
Judy Trogadis Eye Research Institute and University of Toronto Toronto Hospital, Western Div. 399 Bathurst St. Toronto, Canada M5T 2S8
first thank to all the answers to my question how to record video sequences in PAL norm which are coming from a Philips ESEM FEG using the NTSC norm. We are looking to bye a video recorder from AIWA, which costs about 1800DM and which should do what we want. Until now we have no experiences with that system.
Kind Reagards
Rainer Ziel
(Rainer.Ziel-at-Akzo.NL)
Rainer Ziel, Akzo Nobel Central Research, 63784 Obernburg, Germany
some time ago you posted the message below on the microscopy list. I am in the process of buying a similar system, and ran into the same problem. Which solution have you chosen? And what are you experiences? Regards, Willem-Pier Vellinga
Materials Technology, Mechanical Engineering Eindhoven University of Technology
} Dear all, } we want record video sequences on a video tape. Unfortunably the video norm used at the Philips SEM XL30 ESEM FEG is the NTSC-norm, whereas the video norm in Germany is PAL. Are there any experiences using an NTSC to PAL converter? What is about the image quality? Are video recorder available, which can understand the NTSC-norm and writing the video signal in PAL-norm to the video tape and where to get them in Germany? Do you know a software running at Windows NT, which can record video sequences comming from the SEM and which can be stored to hard disc? I would appreciate any answers and comments, especially if you have other ideas to solve the problem. Kind Regard Rainer
I work with a TEM and I'm currently observing and performing EDS analysis on gold samples. The problem is that the samples don't stick to the grids. I need folding grids but the ones I've seen so far are Cu, Ni, Au or Ag.
I need carbon folding grids and would appreciate if anyone can tell me where I can get them.
Isabel Nogueira Dep. Materiais, Instituto Superior T=E9cnico Av. Rovisco Pais, 1 1096 Lisboa Codex PORTUGAL Tel: 351-1-8412120/4 =46ax: 351-1-8412120 e-mail: isabeln-at-alfa.ist.utl.pt
McCrone Research Institute has begun a certification program in Applied Chemical Microscopy. The program has been developed in response to growing student and employer interest in formal individual assessment by the Institute. Employers have been seeeking more specific educational goals, with documented student performance and skill assessment. Students have also been interested in more formal evaluation and documentation of their capabilities in Chemical Microscopy. Certification in Applied Chemical Microscopy is based on (1) successful completion of McCrone sResearch Institute courses satisying certain breadth requirements, (2) Passing of a comprehensive written examination, and (3) Passing of a series of practical proficiency trials. Upon succesful completion of the requirements, candidates will be Certified for their knowledge and ability in Applied Chemical Microscopy. Individuals interested in this certification program should contact: McCrone Research Institute, 2820 S. Michigan Ave, Chicago, IL 60616, Tel.: (312)842-7100, Fax: (312)842-1078, eMail: info-at-mcri.org, www: http://www.mcri.org
Don Grimes, Microscopy Today I have, unfortunately, no interest (financial or otherwise) in the McCrone Research Institute.
Most microscopes with incident fluorescence illumination have some kind of a self-contained filter set or filter cube arrangement. The filter sets have three components: (1) Excitation filter (for proper excitation wavelength); (2) Dichroic mirror (reflects UV into objective, transparent to longer wavelength emission; and (3) Suppression filter (absorbs short wavelength excitation light which is reflected back into the objective, but permits passage of the emission signal).
Try to get some information on the excitation and the emission spectra of your specific acridine dye, if possible. If not, try to match the following:
To excite in the blue range, use an excitation filter in the 450-490nm range. To get into the violet range, use an excitation filter in the neighborhood of 420-490nm. I believe most of the acridine dyes only need violet-blue light for excitation (acridine yellow, acridine orange). There is an acridine blue dye which excites in the UV range at 340-380nm.
The dichroic mirror for most of the acridines should have a cutoff around 510nm; if you use acridine blue (needs UV excitation), the cutoff for the mirror should be around 400nm.
The suppression filter for acridines should be a longpass filter around 515nm. Once again, the exception is acridine blue (UV excitation), which requires a longpass filter around 425nm.
I hope this is of some help. Good luck, and let us know how things work out!
Best regards,
Bob Chiovetti E. Licht Company Leica / Boeckler Instruments / Heidenhain / Narishigi / Colorado Video / Kinetic Systems / Pryor Scientific / Compumotor / Advanced Database Systems / Cohu / Javeline / Optronics / Diagnostic Instruments / Dage MTI / Hitachi / Panasonic / Polaroid / Kodak / Mitsubishi / Sony
Isabel Nogueira wrote: ================================================== I work with a TEM and I'm currently observing and performing EDS analysis on gold samples. The problem is that the samples don't stick to the grids. I need folding grids but the ones I've seen so far are Cu, Ni, Au or Ag.
I need carbon folding grids and would appreciate if anyone can tell me where I can get them. =================================================== I guess I could always be wrong about this, but the only folding grids I have ever seen or heard of are in the above mentioned elements referenced above. Carbon grids are extremely brittle and there would be no such thing as a ductile "hinge" for a carbon grid. That is one of several reasons why folding carbon grids have never been made.
If the goal would be to have a grid that would not contribute x-ray lines, while Be would in theory "work", no one (at least to my knowledge) has ever made a folding Be grid and although technically we could make one, the reality check here would be the extraordinarily high cost to make the first one. And we would have concern about the ultimate ductility of the "hinge" here too.
With regard to your problem, I am surprised that the gold particles, assuming they are less than 30-40 nm in size, are not adhering to the grid. When we have applied gold particles in this size size range to either carbon or Formvar(R) films, we have not had such difficulties. Or I guess I should more correctly say, we have not been aware of any such difficulties being present. Some many years ago we had some non-conductive colloid (MgO) that was not adhering and what solved the problem was to disperse on Formvar, after which we exposed the film to the vapors of some solvent, if I remember correctly, chloroform. This seemed to be just enough to cause the needed adhesion between particle and support film. Hence the final film could be supported with either Be or diamond grids (see our website below for details ), neither of which would contribute anything to the final EDS spectrum.
In any case, much as I would like the opportunity to produce an exotic custom made grid, there might be a more simple and less expensive approach to solving the problem.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
I assume that the x-ray lines from the available folding grids will affect your analysis (for example - you could prove there was no Cu using a gold grid then use Cu grids for your analysis).
I don't think carbon grids will fold. If you have to use C then how about making a sandwich of the specimen between two (aligned) carbon grids and (carefully) glueing the edges of the grids together.
Good Luck Ron
=========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
Dear Isabell - I don't think that oysters grids in carbon exist. Carbon grids are extremely brittle and could not work unless somebody finds a very differe= nt kind of carbon grid. Since you are not trying for truly quantitative work, I would check which metal grid would give least peak overlap. Nylon grids (not available as oysters either) contain a little Ti and they are not rigid enough for mos= t material specimens.=20 Specimens suitable to be held within an oyster type grid are likely to be too thick for TEM unless they are ion beam thinned. In which case, fixing the specimen to a large single hole grid with a bit of superglue is a better alternative.=20 The large single hole option could also avoid almost all grid related X-rays. TEM holder permitting, you could make a sandwich with a second single large hole grid glued on top. For gluing use superglue and a low power dissecting scope. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 500 Links, MSDS, User Notes =20 ************************ http://www.proscitech.com.au
-------------------------------- I work with a TEM and I'm currently observing and performing EDS analysis on gold samples. The problem is that the samples don't stick to the grids. I need folding grids but the ones I've seen so far are Cu, Ni, Au or Ag.
I need carbon folding grids and would appreciate if anyone can tell me where I can get them.
Isabel Nogueira Dep. Materiais, Instituto Superior T=E9cnico Av. Rovisco Pais, 1 1096 Lisboa Codex PORTUGAL Tel: 351-1-8412120/4 Fax: 351-1-8412120 e-mail: isabeln-at-alfa.ist.utl.pt
On 11/26/97 11:45:27 you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Dr. Cornell, We analyze Li-based oxides for stoichiometry by Auger (if thickness of oxide layer is less than 0.5 micron), XPS or SIMS. The later technique is used more for impurity level analysis rather than for stoichiometry. However, the best option is still a chemical analysis as soon as oxide of interest can be removed from substrate without the damage to it. If you have any question do not hesitate to call me at 510-567-0480. Yours, Igor Ivanov Analytical Services RJ Lee Group, Inc. 530 McCormick Str. San Leandro, CA 94577
} Date: Wed, 26 Nov 1997 15:55:56 -0800 } From: Pat Hales {hales-at-medcor.mcgill.ca} } To: microscopy-at-sparc5.microscopy.com } Subject: LKB/Reichert Parts and Service Wanted } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } We have an LKB 2178 Knifemaker II that needs servicing - it takes a lot of } pressure before it will score the glass and then chips the glass edge rather } than a clean fracture. Does anyone know of someone who services these in the } Montreal, Canada area?
SOUNDS LIKE YOU NEED A NEW SCORING WHEEL. WE CHANGE OURS OURSELVES. } } Secondly, we need parts for a Reichert FC4 unit - specifically the upper } part of the cryo knife holder and the specimen (not chuck) set screw and } allen key. Again, does anyone know who sells these to the Montreal area? } TEKNET SERVICES OUR CRYOTOMES IN NC. THEY LIVE IN NJ---PROBABLY ABOUT THE SAME DISTANCE FROM YOU AS US.
1 800 853 6386
} Thanks in advance, } } Pat Hales } McGill University } Dept. of Anatomy & Cell Biology } hales-at-hippo.medcor.mcgill.ca }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
Could you put a formvar film *over* the sample? Maybe by putting the sample onto the film while it is floating on the water, then putting the grid, already coated, onto the sample, then picking up the whole sandwich and cutting round the grid. Just a thought.
Lesley Weston.
On Thu, 27 Nov 1997, Isabel Nogueira wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } =20 } =20 } I work with a TEM and I'm currently observing and performing EDS analysis } on gold samples. } The problem is that the samples don't stick to the grids. } I need folding grids but the ones I've seen so far are Cu, Ni, Au or Ag. } =20 } I need carbon folding grids and would appreciate if anyone can tell me } where I can get them. } =20 } =20 } =20 } Isabel Nogueira } Dep. Materiais, Instituto Superior T=E9cnico } Av. Rovisco Pais, 1 } 1096 Lisboa Codex } PORTUGAL } Tel: 351-1-8412120/4 } Fax: 351-1-8412120 } e-mail: isabeln-at-alfa.ist.utl.pt } =20 } =20 } =20
The technique that I find works well almost always (except on Friday afternoons!) using the colloidal silver liquid adhesive, is to position your specimen first - a fraction off the final position. Then apply a small drop of adhesive using the point of a sewing needle and gently push the edge of the specimen into that drop. At this point you must wait until the glue dries, then apply another two drops of adhesive to form three roughly equally spaced drops.
Hello=20 I remember for sure that few weeks ago somebody asked for contact to = Gernot Winkler whose making refurbishing of=20 old microsonds and similar things in Germany. I was sure that I have somewhere his card - and this is it:
EMS -Mikrosonden Systeme Service GmbH Gernot Winkler Am Steinbruch 2 97859 Wiesthal tel: 06020/97130 fax: 06020/971330 mobile: 0171/5005759
I hope that this message will be recognised by interesant person.
regards=20
Krzysztof M.Herman LabSoft Sp.C. 21 Kosciuszki Str. 05-500 Piaseczno, Poland tel/fax: (48 22) 7502024, 7502028, 7570671 fax: (48 22) 483787, mobile (48 90) 213438 E-mail: labsoft-at-labsoft.com.pl http://www.labsoft.com.pl/
When you say gold samples, are they powders or foils. If you are talking about powders (or dispersions) you will need some sort of support such as a formvar support or a holey grid . You can make this in your lab or purchase them from suppliers. Otherwise, you might try applying an adhesive coating to the grid by dissolving scotch tape in acetone and immersing the C-grids in this solution so that it will coat the grid bars and make them sticky enough.
I work with a TEM and I'm currently observing and performing EDS analysis on gold samples. The problem is that the samples don't stick to the grids. I need folding grids but the ones I've seen so far are Cu, Ni, Au or Ag.
I need carbon folding grids and would appreciate if anyone can tell me where I can get them.
Isabel Nogueira Dep. Materiais, Instituto Superior Tecnico Av. Rovisco Pais, 1 1096 Lisboa Codex PORTUGAL Tel: 351-1-8412120/4 Fax: 351-1-8412120 e-mail: isabeln-at-alfa.ist.utl.pt
We have a Linkham heating/cooling stage that appears to perform according to its specifications. We have not done a rigorous evaluation of the stage temperartures/drift etc. but with the standards we have run it has provided results close to expected values. It has a stated range range of -190 to +600 degrees C. We have used it in the -50 to +200 range with no problems. Overall a good unit but a bit pricey.
Joe Neilly Abbott Labortories Dept. of Microscopy and Microanalysis 200 Abbott Park Rd. Abbott Park, IL 60064
We have attempted to addressed the issue of sample retention in a TEM in a short technical note in Journal of Microscopy, V 174, Pt. 1, April 1994, pp. 55-58, "Enhanced retention of magnetic particles (e.g. microtomed sections) in a TEM". ( E-mail valdemar-at-fast.net with a request for a reprint. )
The technique boils down to fabricating a bi-layer, electron transparent support film on a standard TEM grid of your choice. The materials in the two-layer support film are chosen for solvent incompatibility (e.g. collodion top film soluble in amyl acetate over the bottom layer of Formvar soluble in ethylene dichloride but not in amyl acetate); so that, after placing your sample on the top film, the top film is temporarily softened and transformed into electron transparent layer of adhesive with a judicious application ( a few drops to wet filter paper near your TEM grid ) of solvent specific to that film. The bi-layer support films are a lot easier to fabricate than the description makes it appear, and the technique is amenable to any combination of support films as long as the structural layer is not affected by the solvent for the adhesive layer.
Prior to developing this method, we had a severe problem with losses of microtomed sections of magnetic steel in the field of a high excitation objective lens. Now, we hardly ever loose one; and the bi-layer support films are sufficiently robust for sample deposition with a hand-held eye lash, and sufficiently stable and clean for multi-hour acquisition of quantitative composition profiles at high resolution by EDS under UHV in a dedicated STEM. Perhaps with an exception of some EELS work, I see no reason why the technique would not work under most circumstances for your persnickety sample with a judicious selection of the support films and their solvents.
We have tried the folding grid approach and, by comparison, found it a bother.
For a method of applying an adhesive to grid bars, you might look up E. Fritz (1991), "The use of adhesive-coated grids for the X-ray microanalysis of dry-cut sections in the TEM", J. Microsc. 161, 501-504. ( Sorry, but I'm out of copies of this one. )
Valdemar Furdanowicz valdemar-at-fast.net Homer Research Labs Bethlehem Steel Co. Bethlehem, PA 18016
******************************************************** Isabel Nogueira wrote: ----------------------------------------------------------------------- I work with a TEM and I'm currently observing and performing EDS analysis on gold samples. The problem is that the samples don't stick to the grids. I need folding grids but the ones I've seen so far are Cu, Ni, Au or Ag.
I need carbon folding grids and would appreciate if anyone can tell me where I can get them. -----------------------------------------------------------------------
Nancy Buening wrote: ----------------------------------------------------------------------- I am analyzing calcite shells in the TEM. I am having difficulty gluing the calcite samples onto the grid. I have been using colloidal silver liquid, but it either dries before I can get the calcite in place or I get too much liquid and the grid "drowns". Is this just a matter of practice, or is there another gluing material that I can try that is more forgiving ? -----------------------------------------------------------------------
Does anyone have any simple solutions to reduce the amount of endogenous fluorescence that seems to be both tissue type and fixation specific? We work with fish retinas and as a rule immersion fix in 4% paraf. for 1 hr, room temp. When glut is added the problem is worse. The fluorescence seems to be worse in the inner segments of the photoreceptors-all these years the results have been satisfactory, but any improvement would be great. Linda Barthel Research Associate II Department of Anatomy and Cell Biology University of Michigan lab (313) 764-7476 fax (313) 763-1166 barthel-at-umich.edu
I am trying to serial section some fungal spores for TEM, and I am running into problems with the sections wrinkling.
Details: I am using formvar and carbon coated slot grids. The specimen is embedded in epon. I am using a Reichert Ultracut E (mechanical advance)
ultramicrotome, and an Edgecraft diamond knife. I have been moving the sections onto the grids with the aid of a micromanipulator holding the grid into the water, touching the section ribbon to the formvar, and winding the grid up out of the water.
I have tried varying the section size - from about 1/2mm*1/2mm to less than 1/4mm each side, with little or no change in the degree of wrinkling. I tried varying the angle at which the grid is held into the water, from almost parallel, to perpinduclar, with no success. I tried varying the speed at whcih the grid is removed from the water, and hence the ribbon is sucked onto the formvar, also with no success. Finally, I thought it might be the formvar/carbon coat -- when held close to the boat, the humidity caused the film inside the slot to go slack. I thought maybe the slight waviness in the film might be introducing the wrinkles, so I changed my formvar recipe and procedure. I did manage to produce a couple grids with intact films which did not slacken near the boat, but the sections still wrinkled. The sections are uniform silver/gold interface, and are generally part of a very strong ribbon (can be hard to break it up into managable pieces using eyebrow brushes).
So, here I am: does anyone have any suggestions for avoiding the %&*%$$$# wrinkles, other than changing the embedding medium properties?
Also, has anyone else ever noticed the slackening of the formvar film near to water, what might cause it, and how best to avoid it? (my good grids were made with formvar that was thoroughly dessicated in dichloroethane - I was previously using chloroform without the variety of drying procedures).
Thanks in advance (from me and my supervisor whoe would appreciate it if I didn't throw the ultramicrotome through the window),
Russell
PS I have also tried picking up the sections with an empty slot grid and overlaying that grid over another, coated grid, and allowing the sections to dry. They still wrinkled. R.
I have a Durst Laborator S-45-EM and the varipoint unit that adjusts light intensity has broken. Does any one have any idea how to fix or replace this unit. I don't expect that a new replacement is available but maybe someone has a Laborator that they are no longer using can provide this part. Or maybe some one can tell me how to build a unit that will accomplish the same function so that I can continue to use the Laborator.
Also can anyone tell me how to find Steve Miller who used to be with Integrated Microsystems
Bob Schmitz Robert J. Schmitz Electron Microscope Lab Department of Biology, CNR Building University of Wisconsin Stevens Point Stevens Point, WI 54481 phone (715) 346-2420 FAX (715)346-3624
I know how you feel. Be patient. I have been doing serial sectioning for 6 years. We have an elaborate set up which allows us to section in pairs. One person sections and one person transfers the sections to the formvar coated slot grid. If you want to know about this transfer apparatus I can find the reference for you.
In the meantime you will have to work by yourself. Whenever I section by myself I get some wrinkles(nothing to upset research though). The wrinkles occur as I slide the formvar coated slot grid into the diamond knife water, and draw up the sections. The wrinkles are the result of the formvar stretching a little when it goes under water. Then, as the sections lie down on the stretched formvar, they wrinkle a bit. I never have wrinkles which are bad enough to stop science.
To minimize wrinkles I suggest: Make your block face as tiny as possible.
Immerse your grid at a 45 degree angle.
Slowly coax your sections onto the formvar, and slowly and gently pull the grid out of the water at a 45 degree angle. The key is to move slowly and deliberately.
Work on a day that you are happy, the block is happy, the sectioning room is happy. Do not attempt EM on a bad hair day.
Cleaning the ol' closets. Before I throw these in the dumpster, I thought I'd ask: JEOL 100B specimen carousel and specimen holders JEOL 100B specimen exchange mechanism (parts) 2-3 boxes of HU11E replacement parts and vacuum tubes EDAX detector and 183 preamp, from AMRAY SEM. Broken window, maybe other damage
You pay to ship it, it's yours. Contact me at the address below:
Julian P.S. Smith III Biology Winthrop University Rock Hill, SC 29733 803-323-2111 x227 (vox) 803-323-2246 (fax)
jd wrote: } } It has been a few years since I have been involved with temperature } control stages on optical microscopes. Last I remember there were } issues such as surface area temperature mapping(variations) functions, } isolation, target range overshooting, steady state vs. drift } functions, heating and cooling techniques, depth of focus and other } optical issues, maximum and minimum feasible temperatures, heating and } cooling techniques, etc. Can anyone refresh my memory , bring me up to } date ,and refer me to journal and review articles on the subject. } I would like references to current manufacturers. Also I would be } interested in personal experiences with specific current equipment } manufacturers of both microscopes and control stages for this use } (preferably any potentially negative feedback opinions sent only to my } personal email address below, so no manufacturer is embarrassed or } damaged by personal opinions in this forum/possibly an obvious point } of controversy). } Finally does anyone have any personal experience with modifications to } existing designs or references to in house construction of these } devices. } I also might be prospect for use equipment of this nature. } } JD } EMAIL1: wa5ekh-at-juno.com } please cc to EMAIL2: wa5ekh-at-cyberramp.net Dear JD,
There are several people who make very good heating stages. Yes, there are issues such as surface area, heating and cooling techniques, etc. Typically, however, the better stages have fairly enclosed heating chambers, so there is less concern about heat transfer. A few quick cautions: you probably will have to get long working distance objectives, especially if your application requires higher magnifications. Also,you have not mentioned much about the specifics of your work, so it is hard to answer in a more focused fashion, but you may not be able to look at a very extensive area at any given time.
I have had personal experience with two manufacturers: Mettler and the Koffler hot stage available through Arthur Little. Mettler's stages are well built and, depending on the options purchased, can provide extremely delicate temperature control and sophisticated heating and cooling cycles. They are also very expensive. The Koffler hot stages are much more practical and allow you to look at larger samples. As I remember it, they had an interesting large glass plate which covered the chamber and kept the temperature fairly constant. Both typically have a range from some sub-room temperature value to about 350 degrees C.
There are three other companies which I would suggest that you investigate as well: Bioptechs, PhysiTemp, and World Precision Instruments. All three are better suited to the 32-35 degree C, constant temperature conditions necessary for live cell work. I have only seen brochures for the PhysiTemp but I have had some lengthy conversations with the people at Bioptechs. If you are doing critical live cell work, they have a number of options whichhallow you to look at somewhat larger fields. Also, for things like Calcium flux test, where even the temperature of the objective lens might be critical, they have an objective heater.
Finally, on the do-it-yourself front: Walter McCrone runs courses on hot stage microscopy out of the McCrone Institute in Chicago. Part of the class revolves around using a rheostat, wires, and thermally conductive glass to build your own stage. The good news is that the stage has a very low profile so that you can look at larger areas and don't need extra long working distance objectives. Secondly, they are very inexpensive to build. However, unless you have experience with thermocouples, I don't know how you would accurately measure the tempature at the sample. Also, the sample is open to ambient conditions. I am not quite sure how Walter's design accounts for air currents, local cooling, etc. I have worked with a client who built one of these devices for pigment and polymer analysis and he seemed quite happy with it. (I will attach names and addresses for any contacts which I have in my files, below).
Microscopy/Microscopy Education tries to act as a clearinghouse for this type of information, so we would really appreciate a summary of your findings.
Hope this all helps.
Barbara Foster Consortium President MME 53 Eton Street Springfield, MA 01108 US PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com
America's first consortium of expert microscopists dedicated to helping you be more productive with your microscope.
DISCLAIMER: MME does not sell or have any financial interest in any of the above mentioned equipment.
List of contacts: Bioptechs - Dan Focht (412)282-7145 Web site: www.Bioptechs.com Mettler - Mark Kelsey (800)638-8537 ext 7041 McCrone Institute - ask for any of their fine technical staff - (312)842-7100
PhysiTemp advertises in most of the common trade journals. I will have to send you both their information and Arthur Little's under separate cover.
You could try waving a Q-tip soaked in chloroform over the sections before bringing the grid anywhere near them (this is not the same as waving a dead chicken - it works). Or you could buy a heat-pen from (I think) JB EM, and probably other suppliers. In this case, you wave a hot wire over the sections and they flatten like magic. Hope this helps.
Lesley Weston.
On Mon, 1 Dec 1997, Wyeth, Russell wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } TWIMC, } } I am trying to serial section some fungal spores for TEM, and I am } running into } problems with the sections wrinkling. } } Details: I am using formvar and carbon coated slot grids. The specimen } is } embedded in epon. I am using a Reichert Ultracut E (mechanical advance) } } ultramicrotome, and an Edgecraft diamond knife. I have been moving the } sections onto the grids with the aid of a micromanipulator holding the } grid } into the water, touching the section ribbon to the formvar, and winding } the } grid up out of the water. } } I have tried varying the section size - from about 1/2mm*1/2mm to less } than } 1/4mm each side, with little or no change in the degree of wrinkling. I } tried } varying the angle at which the grid is held into the water, from almost } parallel, to perpinduclar, with no success. I tried varying the speed } at whcih } the grid is removed from the water, and hence the ribbon is sucked onto } the } formvar, also with no success. Finally, I thought it might be the } formvar/carbon coat -- when held close to the boat, the humidity caused } the } film inside the slot to go slack. I thought maybe the slight waviness in } the } film might be introducing the wrinkles, so I changed my formvar recipe } and } procedure. I did manage to produce a couple grids with intact films } which did } not slacken near the boat, but the sections still wrinkled. The } sections are } uniform silver/gold interface, and are generally part of a very strong } ribbon } (can be hard to break it up into managable pieces using eyebrow } brushes). } } So, here I am: does anyone have any suggestions for avoiding the } %&*%$$$# } wrinkles, other than changing the embedding medium properties? } } Also, has anyone else ever noticed the slackening of the formvar film } near to } water, what might cause it, and how best to avoid it? (my good grids } were made } with formvar that was thoroughly dessicated in dichloroethane - I was } previously using chloroform without the variety of drying procedures). } } Thanks in advance (from me and my supervisor whoe would appreciate it if } I } didn't throw the ultramicrotome through the window), } } Russell } } PS I have also tried picking up the sections with an empty slot grid and } overlaying that grid over another, coated grid, and allowing the } sections to dry. They still wrinkled. R. } }
Bob - Find a place through your yellow pages which rewinds electric motors. Phone them and ask if they can rewind a variable transformer. Otherwise, buy a variable transformer with similar specifications and adapt it to the equipment. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au
---------- } } I have a Durst Laborator S-45-EM and the varipoint unit that adjusts } light intensity has broken. Does any one have any idea how to fix or } replace this unit. I don't expect that a new replacement is available } but maybe someone has a Laborator that they are no longer using can } provide this part. Or maybe some one can tell me how to build a unit } that will accomplish the same function so that I can continue to use the } Laborator. } } Also can anyone tell me how to find Steve Miller who used to be with } Integrated Microsystems } } Bob Schmitz } Robert J. Schmitz } Electron Microscope Lab } Department of Biology, CNR Building } University of Wisconsin Stevens Point } Stevens Point, WI 54481 } phone (715) 346-2420 } FAX (715)346-3624
} From: "jd" {wa5ekh-at-cyberramp.net} } To: {microscopy-at-sparc5.microscopy.com} } Subject: Temperature Control Stages for Optical Microscopy } Date sent: Fri, 21 Nov 1997 03:39:31 -0800
I sort of feel that people using this list should reveal at least their name, if not their affiliation. I don't want to start anything both big and trivial, but what do others think?
Ritchie
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
} I have a Durst Laborator S-45-EM and the varipoint unit that adjusts } light intensity has broken. Does any one have any idea how to fix or } replace this unit. I don't expect that a new replacement is available } but maybe someone has a Laborator that they are no longer using can } provide this part. Or maybe some one can tell me how to build a unit } that will accomplish the same function so that I can continue to use the } Laborator.
Bob,
I'm assuming that you're talking about the potentiometer that controls the point light source. I AM NOT an electronics whiz, so I may be completely off base, but I'm pretty sure that those units are nothing more than expensive, heavy-duty "dimmer switches", combined with a voltage transformer. Check with an electronics shop who might be able to fix it fairly easily---I can't imagine that they can be too complicated. If it can't be fixed, I'd bet that a substitute could be made and perhaps fitted with the connectors on the unit you have. It might even be that a hardware store with a good electrical section might have a transformer and dimmer switch with specifications compatible with your light source.
One caution: I would NOT---repeat, NOT---try to use just a dimmer switch hooked to a regular 110-120 outlet. I once had just finished changing from conventional tungsten light to the point light source, grabbed the wrong cord and plugged it directly into an outlet. When I turned on the switch, the bulb exploded like a pistol going off and sprayed half the darkroom with hot shards of glass (I hadn't closed the enlarger head access panel, yet). Get a good electrician's advice before rigging a substitute, but I bet it can be done.
Randy Tindall Electron Microscope Lab New Mexico State University Las Cruces, NM 88003 Randy Tindall 2017 Princess Jeanne Las Cruces, New Mexico 88001-4157
{bold} {fontfamily} {param} Times {/param} {bigger} POSTDOCTORAL POSITIONS IN MATERIALS SCIENCE
OAK RIDGE NATIONAL LABORATORY
METALS AND CERAMICS DIVISION
{/bigger} {/fontfamily} {/bold} {fontfamily} {param} Times {/param} {bigger} Postdoc= toral positions are available at Oak Ridge National Laboratory (ORNL) in the Metals and Ceramics Division. The positions are in association with the Microscopy and Microanalytical Sciences Group at ORNL The candidate(s) should have proven expertise in electron microscopy of materials, including CTEM and associated defect analysis techniques, as well as HREM. Demonstrated experience in analytical electron microscopy techniques such as high spatial resolution energy dispersive and electron energy loss spectroscopy and the associated techniques of energy-filtered imaging and spectrum imaging is desired. Two postdoctoral positions are available and are described below:
(1) This position is with the Structural Materials Group. The group is participating in national and international programs involved in the development of materials for structural applications in near-term and long-term fusion energy systems. The research is in three separate but related tasks: =20
(a) characterization of the precipitation behavior of oxides, carbides, nitrides, and silicides in suppport of the development of improved alloys in the V-Cr-Ti system.
(b) segregation. precipitation, and evolution of the damage structure in neutron irradiated V-Cr-Ti alloys, and
(c) phase stability, interfacial segregation, and the distribution of helium in neutron irradiated ferritic-martensitic steels.
Experience in the areas of radiation effects and microstructure-property correlation is preferred.=20
(2) This position is in the Microscopy and Microanalytical Sciences Group. The work involves examination of the microstructure and microchemistry of oxide scales which form on advanced materials including intermetallic alloys, superalloys and related alloys.=20 Included in this effort are general microstructural and microchemical characterization of the base material and the oxide scale morphology by SEM and TEM techniques, as well as high spatial resolution microanalysis of boundary segregation to the metal/oxide and oxide/oxide interfaces. This work will involve interactions with the Corrosion Science and Technology Group at ORNL. Experience in high-temperature corrosion is desirable but not required.
The Microscopy and Microanalytical Sciences Group has a complete suite of analytical electron microscopes including: Philips CM12T/STEM 120kV AEM with light-element EDS ; Philips CM30T/STEM 300 kV AEM with EDS and Gatan 678 Imaging Filter (GIF); and a Philips CM200FEG/STEM 200 kV with EDS detector, PEELS and spectrum imaging capability. Also available is Philips XL30 FEG-SEM with back-scattered detector, light-element EDS, Electron Backscattered Pattern (EBSP) camera for localized orientation mapping of materials and wavelength-dispersive spectrometer (WDS) for trace element detectabilty.
Please mail applications (electronic applications will not be accepted) to:
Kathleen B. Alexander
Group Leader, Microscopy and Microanalytical Sciences Group
I recently came across an article where someone was using an "Air Stream Incubator" from Nevtek as a temperature controler. It is apparently a blower that moves heated air across the stage area heating everything in its path. I have never heard of this before. Any comments on the effectiveness of this solution? Dave Knecht
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut U-125 Storrs, CT 06269 Knecht-at-uconnvm.uconn.edu
1. Any glut will cause autofluorescence, in fact we use it as counterstain if we want the tissue to be fluorescent.
2. I remember that someone was using .05% Pontamine Sky Blue in PBS with 1% DMSO after the immunohistochemistry was carried out, to reduce the autofluorescence in the FITC channel.
3. Another person was using .3g eriochrome black in 100ml PBS as a counterstain for the same purpose.
Hope this helps,
Bob
On Mon, 1 Dec 1997, Linda Barthel wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Does anyone have any simple solutions to reduce the amount of endogenous } fluorescence that seems to be both tissue type and fixation specific? We } work with fish retinas and as a rule immersion fix in 4% paraf. for 1 hr, } room temp. When glut is added the problem is worse. The fluorescence } seems to be worse in the inner segments of the photoreceptors-all these } years the results have been satisfactory, but any improvement would be } great. } Linda Barthel } Research Associate II } Department of Anatomy and Cell Biology } University of Michigan } lab (313) 764-7476 } fax (313) 763-1166 } barthel-at-umich.edu } } } }
Thanks for the overwhelming response! Could this possibly a reaction to a common, and all to evil foe?
Here's a quick (sort of) summary for everyone's benefit, and I'll try to answer all the further questions I received.
First, thank you everyone for the chloroform suggestions, but that's not the problem here. I've used that method many-a-time for wrinkled sections coming off the knife blade. These sections, though, are beautifully flat -- they shouldn't require any stretching. The wrinkles are _definitelly_ introduced when the sections attach to the formvar (I carefully checked the grids at all stages). There's just 3 or 4 per section, and they appear no matter whether they are left to dry down onto the formvar from another uncoated grid, or attached directly on to the formvar. (Someone pointed out this shouldn't get in the way of the observations, and in this case - with lots of spores in each sample - the wrinkles are not a critical problem. But I'm a perfectionist, and I can certainly envisage times when I wouldn't want any wrinkles, so I figured I'd try to learn with this otherwise very cooperative sample.)
A good suggestion was made regarding refinement of the micromanipulator technique I already use -- I'll give that one a shot first, but I have a feeling my perfectionist attitude towards the wrinkles may mean I'll need to try something else. The other suggestions centered around alternative means to get the sections on the grid. Basically, the sections are picked up on a formvar film attached to another tool with a hole big enough for the entire film to be passed over onto a clean slot grid. This means the sections can be positioned much more precisely over the slot. (I had heard of this method, but we are cash strapped and I was hoping to avoid a purchase of the loops or 'domino racks'. Perhaps it is unavoidable.) I was also given a couple references which I am about to go look up. I've attached the originals of various suggestions below.
Finally, everyone advocated patience. I was going to say I'd already tried that, but I guess I'll have to go out and buy some more.
Once again, thanks everyone, and I'll report back.
Russell
} snip I use domino racks that can be purchased from the EM companies like EMS or Ted Pella. Instead of working with the formvar on the grid you put the film over the domino rack which is a piece of sheet metal with slightly larger than grid size holes. You then pick up the sections from the water with a grid and don't have to worry about the film wrinkling when you get close to the water. The grids are standard cleaned slot grids. After touch down to the water the sections should be held in the center of the hole by surface tension. You then lay the grid on the hole of the domino rack and let it dry overnight. After the sections have dried, you carefully punch out the grid by using the tips of the forceps around the outside of the grid. I find that this approach works much better for me.
Patty Jansma
} snip I have been cutting ultrathin sections for a number of years now and the most recent advance in section collection to me was the invention of the 'Perfect Loop'. With this loop one can pick up sections from the water bath and place them on a grid sitting on filter paper by the side of the ultramicrotome. The excess water is then wicked away with a wedge shape of filter paper. To date, this I find is the best way to collect sections without wrinkles. This is sold in the USA by EMS. Happy sectioning. Ian Lamswood
} snip Whenever I section by myself I get some wrinkles(nothing to upset research though). The wrinkles occur as I slide the formvar coated slot grid into the diamond knife water, and draw up the sections. The wrinkles are the result of the formvar stretching a little when it goes under water. Then, as the sections lie down on the stretched formvar, they wrinkle a bit. I never have wrinkles which are bad enough to stop science.
To minimize wrinkles I suggest: Make your block face as tiny as possible. Immerse your grid at a 45 degree angle. Slowly coax your sections onto the formvar, and slowly and gently pull the grid out of the water at a 45 degree angle. The key is to move slowly and deliberately. Sally Shrom
} snip I did serial section reconstructions of flagellar apparatuses of two algae for my dissertation work using polystyrene films (Journal of Electron Microscopy Technique 13:268-269. The technique is a modification of the following: Rowley III, J.C. and Moran, D.T. 1975. A simple procedure for mounting wrinkle-free sections on formar-coated slot grids. Ultramicroscopy 1:151-155. I highly recommend this way. It's very easy - I taught it to several persons and they all became better at it than I am. Good luck, Heather Owen
} one final snip A suggestion: I was once told that formvar is less hydrophobic if you refrigerate the grids overnight before use. I never needed to, but it was suggested. Otherwise, good luck. Serial sectioning is no fun trick. Gregg Sobocinski
EXXON POSTDOCTORAL POSITION IN ELECTRON MICROSCOPY AND MATERIALS SCIENCE OF CATALYTIC MATERIALS
Corporate Research Laboratory Exxon Research and Engineering Company
Exxon Research and Engineering Company in Annandale, New Jersey has an immediate opening for a postdoctoral fellow in electron microscopy. This fundamental research position will explore the limits of field- emission TEM and STEM of ultrafine metal particles. Our work involves quantitative high resolution imaging, x-ray mapping, and electron holography of nanoscale metal particles on model supports or within commercial catalysts. Depending on the interests and expertise of the candidate, this work may involve zeolite structure determination and reaction cell treatment of supported metal catalysts. Our primary instrument is a Philips CM200FEG equipped with CCD camera, STEM, PGT EDS, rotatable biprism for electron holography, oil-free high vacuum system, and high resolution objective lens system.
Requirements for this position include expertise with high resolution electron microscopy, electron holography, field-emission analytical microscopy, and scattering physics. Experience with the materials science of catalysts is desirable. Research in this area involves a multidisciplinary team approach which provides an excellent learning environment. Strong communication skills are required for working in our team environment.
The term of the position is one year beginning early in 1998. Extension to two years is likely. Our research lab in rural New Jersey offers convenient access to Philadelphia, Princeton and New York City. Exxon offers an excellent working environment, salary commensurate with skills and experience, and excellent benefits.
Applicants for this position who meet the majority of the qualifications outlined above should forward a resume, publication list and two or three references to:
Dr. Mark M. Disko Corporate Research Laboratory Exxon Research and Engineering Company 1545 Route 22 East Annandale, New Jersey 08801
(908) 730-2503 FAX (908) 730-3314
Please do not respond directly to this list server. In the event you need further information rapidly, forward electronic mail to me at mmdisko-at-erenj.com .
In a message dated 97-12-02 10:14:08 EST, rwyeth-at-pfc.forestry.ca writes:
{ { I am trying to serial section some fungal spores for TEM, and I am running into problems with the sections wrinkling.
Details: I am using formvar and carbon coated slot grids... } }
Russell,
I have encountered similar problems in one of my past lives as an ultramicrotomist, and here's what we came up with:
We found the major contributor was the hydrophobic nature of the carbon film. Even storing the grids under UV light and glow-discharging in a partial vacuum with an air atmosphere really didn't help that much.
We finally collected the sections onto slot grids which had a *formvar only* film, wicked away the excess fluid from the side of the grid, let everything dry well, then did our staining. Immediately before going to the microscope, we placed the grids into a bell jar and gave them a light dusting of carbon. This helped immensely.
Good luck; let us know how things work out in the end!
} } Winter Workshop on In-Situ Electron Microscopy } } A workshop on In-Situ Electron Microscopy will be held in Scottsdale, } Arizona from Jan. 7-10, 1998. The goal of this workshop is to provide a } forum for presentations and discussion of recent developments in } instrumentation, current applications and future directions of in-situ TEM } and SEM. Some areas of particular interest are: } } . In-Situ Heating Experiments: Electron Diffraction and Imaging or } Temperature Controlled Experiments. } . Ion Implantation studies and ion irradiation effects } . Environmental Cells or Gaseous Environment Controlled TEM/SEM } . Effects of Stress/Strain including fracture studies and stress-induced } phase transformation } . Magnetic Materials Studies } } The list of invited speakers includes: } } Ernst Bauer, Arizona State University } Ed Boyes, Dupont Corp. } Kazuo Furuya, National Research Institute for Metals } James Howe, University of Virginia } Robert Hull, University of Virginia } T. Kamino, Hitachi Instruments Eng. Co. } John Mansfield, University of Michigan } Ulrich Messerschmidt, Max Planck Institute of Microstructure Physics } Amanda K. Petford-Long, University of Oxford } Francis M. Ross, IBM Thomas J. Watson Labs } Robert Sinclair, Stanford University } Nubuo Tanaka, Nagoya University } } Abstracts are still being accepted and should be sent as soon as possible. } For more information contact: } } Eloise Kadri } Center for Solid State Science } Arizona State University } P.O. Box 871704 } Tempe, AZ 85287-1704, USA } } Email: Eloise.Kadri-at-asu.edu } Tel: 602 965 9004 } } You can also register via our Web Site at: } http://www.asu.edu/clas/csss/workshop/ } } } } Peter A. Crozier } } Industrial Associates Program } Center for Solid State Science } Arizona State University } Tel: 602 965 2934 } Fax: 602 965 9004 } } Website: http://www.asu.edu/clas/csss/IAP/
John C. Wheatley Lab Manager Arizona State University Center for Solid State Science PSA-213 BOX 871704 Tempe, AZ 85287-1704
} } Winter Workshop on In-Situ Electron Microscopy } } A workshop on In-Situ Electron Microscopy will be held in Scottsdale, } Arizona from Jan. 7-10, 1998. The goal of this workshop is to provide a } forum for presentations and discussion of recent developments in } instrumentation, current applications and future directions of in-situ TEM } and SEM. Some areas of particular interest are: } } . In-Situ Heating Experiments: Electron Diffraction and Imaging or } Temperature Controlled Experiments. } . Ion Implantation studies and ion irradiation effects } . Environmental Cells or Gaseous Environment Controlled TEM/SEM } . Effects of Stress/Strain including fracture studies and stress-induced } phase transformation } . Magnetic Materials Studies } } The list of invited speakers includes: } } Ernst Bauer, Arizona State University } Ed Boyes, Dupont Corp. } Kazuo Furuya, National Research Institute for Metals } James Howe, University of Virginia } Robert Hull, University of Virginia } T. Kamino, Hitachi Instruments Eng. Co. } John Mansfield, University of Michigan } Ulrich Messerschmidt, Max Planck Institute of Microstructure Physics } Amanda K. Petford-Long, University of Oxford } Francis M. Ross, IBM Thomas J. Watson Labs } Robert Sinclair, Stanford University } Nubuo Tanaka, Nagoya University } } Abstracts are still being accepted and should be sent as soon as possible. } For more information contact: } } Eloise Kadri } Center for Solid State Science } Arizona State University } P.O. Box 871704 } Tempe, AZ 85287-1704, USA } } Email: Eloise.Kadri-at-asu.edu } Tel: 602 965 9004 } } You can also register via our Web Site at: } http://www.asu.edu/clas/csss/workshop/ } } } } Peter A. Crozier } } Industrial Associates Program } Center for Solid State Science } Arizona State University } Tel: 602 965 2934 } Fax: 602 965 9004 } } Website: http://www.asu.edu/clas/csss/IAP/
John C. Wheatley Lab Manager Arizona State University Center for Solid State Science PSA-213 BOX 871704 Tempe, AZ 85287-1704
Rick Felten 12/02/97 03:06 PM I am looking for some Pyrope Garnet, Mg3Al2Si3O12, to use as a X-Ray Microanalysis Standard. I have tried many of the EM suppliers (Energy Beam Sciences, Polysciences, Fullam, and Ted Pella) w/o any luck. I would be amenable to another standard that has major levels of Mg, Si and O and a minor level of Al and no other elements. Any suggestions?
} Does anyone have any simple solutions to reduce the amount of endogenous } fluorescence that seems to be both tissue type and fixation specific? We } work with fish retinas and as a rule immersion fix in 4% paraf. for 1 hr, } room temp. When glut is added the problem is worse. The fluorescence } seems to be worse in the inner segments of the photoreceptors-all these } years the results have been satisfactory, but any improvement would be } great.
This problem was recently discussed here. The fluorescence is due to the aldehydes and the fix [:-)] is to reduce them with NaBH4. I am definitely not competent in specimen preparation, but somewhere in our group the procedure has been worked out. If none of the experts in this field respond, I'll try to get the recipe for you. Yours, Bill Tivol
Dear Ritchie, } } I sort of feel that people using this list should reveal at least } their name, if not their affiliation. } I don't want to start anything both big and trivial, but what do } others think? } As one who has first met many people electronically through this list and then seen them at MSA meetings, I think it's useful to put names on postings. I don't think that "jd" had anything sinister in mind, and I certainly would not want Nestor to insist that all postings meet any requirements other than those of relevance, etc. already in place. Yours, Bill Tivol
A friend of mine wants to sell two microscopes. Please contact him directly.
Lietz Dialux Microscope, trinocular head with phototube (modified Orthomat System for SLR camera. Bright field objectives 4x, 10x, 25x (npl), 40x plan, 100x. Two .9 na condensers (brightfield and darkfield). Asking $2,000.
Leitz Diavert Microscope. Mint condition foruse in tissue culture. Phase contrast 10x, 20x, 4x low power objective, 12V 100w light source. Asking $1500 ($2,000 with transformer).
For more information call 614 688 4471 and ask for Steve.
Rick Felten wrote: ================================================ I am looking for some Pyrope Garnet, Mg3Al2Si3O12, to use as a X-Ray Microanalysis Standard. I have tried many of the EM suppliers (Energy Beam Sciences, Polysciences, Fullam, and Ted Pella) w/o any luck. I would be amenable to another standard that has major levels of Mg, Si and O and a minor level of Al and no other elements. Any suggestions? ================================================ Actually you don't have to settle for anything other than what you want! The pyrope garnet, often times requested, is mineral #37 on the SPI #02753 Mineral Mount. It has excellent homogeneity and has been offered by our firm since about 1980. More details about this mount, including 52 other minerals, are available on our website given below. Contact me off line if you have any questions.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
You may have tried borohydride. The following is a brief writeup of how we have used it in the past. A. Kent Christensen, University of Michigan.
BOROHYDRIDE
If glutaraldehyde was used in the fixative, then it may be advantageous to quench extraneous aldehyde groups with 1% sodium borohydride (NaBH4) (Eldred et al., 1983, J Histochem Cytochem 31:285), which is a particularly strong reducing agent (USE WITH CAUTION). Borohydride can eliminate most tissue autofluorescence, thus reducing background in LM ICC studies involving fluorescent markers. It has been suggested that borohydride may partially restore antigenicity after glutaraldehyde fixation by reducing Schiff bases (carbon-nitrogen double bonds) that can be formed when glutaraldehyde reacts with free amino groups on proteins; the reduced bonds are less rigid, and the increased mobility may restore some of the antigenicity.
Borohydride treatment can be carried out on pieces of tissue after fixation and the overnight buffer wash (Eldred et al, 1983). Use 1% sodium borohydride in PBS for 30 min at room temperature. The solution should be freshly prepared from sodium borohydride powder that has previously been stored in a manner that has protected it from moisture. The tissues will bubble vigorously as hydrogen gas leaves them, which will worry you but doesn't seem to damage the tissues. The treatment is followed by a wash of 2 x 30 min in PBS.
To use borohydride as a quenching agent during an LM immunocytochemical run, put a drop of 1% sodium borohydride on each tissue section and leave it for about 10 minutes. Then wash in a coplin jar of PBS or PBS-G.
For convenience, 10 mg aliquots of sodium borohydride powder can be stored in microtubes at -20=A1C in a plastic box containing silica gel desiccant. When you need 1% borohydride, add 1 ml of PBS to a tube and vortex briefly (CAUTION: open the microtube promptly after vortexing, or the hyrogen gas generated as the powder goes into solution will pop it open, possibly causing a spill).
Wrick: Shop around. No problem getting that standard; for one, it is in our catalogue. Note the A$ is 40% less than the US$. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au
---------- } From: Rick Feltenby way of Nestor J. Zaluzec {rfelten-at-Macdermid.com} } To: microscopy-at-sparc5.microscopy.com } Subject: Pyrope Garnet } Date: Wednesday, 3 December 1997 9:15 } Rick Felten } 12/02/97 03:06 PM } I am looking for some Pyrope Garnet, Mg3Al2Si3O12, to use as a X-Ray } Microanalysis Standard. I have tried many of the EM suppliers (Energy Beam } Sciences, Polysciences, Fullam, and Ted Pella) w/o any luck. I would be } amenable to another standard that has major levels of Mg, Si and O and a } minor level of Al and no other elements. Any suggestions? } }
Three announcements from the San Francisco Microscopical Society:
First, we have established an email list of correspondents who wish to receive periodic announcements of our activities, significant updates to our web pages, and other notes of the Society. To be placed on this list, respond to me personally (not the MSA List!) via email, or respond through the solicitation on the first of our web pages (see below).
Second, we would like to invite all friends of our Society to join us at our annual Christmas Social Hour on Thursday evening, December 11, at 7:00 PM at the Rockridge Branch of the Oakland Public Library. Further information can be obtained at our web pages (again, see below).
Third, we would like to announce the establishment of the San Francisco Microscopical Society Web Pages. While in development they will reside at:
After a shaking out period and filling in some gaps we'll probably move to a more permanent URL, but for now I've posted the pages adjacent to my personal pages.
With regard to the web pages, we are curious to know if commercial firms would be interested in a corporate membership which would support the society (we are a CA non-profit corporation) as well as providing an opportunity for exposure to our members through our regular newsletter and to the broader microscopy community through our web pages. If you're with such a firm, please contact me off list and let me know if such a membership would be of interest to your firm. I would anticipate a very modest fee for such a membership.
Fourth, and perhaps most importantly, may you all have a safe, happy, and full holiday season!
Steve Shaffer -- ********************************************************** Stephen A. Shaffer sshaffer-at-microdataware.com MicroDataware http:www.microdataware.com (Under reconstruction and temporarily out of service) Personal stuff: steve_shaffer-at-compuserve.com http://ourworld.compuserve.com/homepages/steve_shaffer/ **********************************************************
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com -----------------------------------------------------------------------.
Hi Rick.
Micro-Analysis Consultants Ltd has been in the business of supplying standards for micro analysis for 16 years now. We have over 300 different materials in stock with certificates of analysis. We are an ISO 9002 registered company for the supply of standards, advertise regularly in Microscopy and Analysis, and have a web site at:
http://www.macstandards.co.uk/town/street/yr49/
We specialise in the supply of full custom standards. In other words tell us what you want and we will quote you for it. You can have whatever block size you would like in either Stainless Steel, Brass, Aluminium or Carbon resin. The number of standards we can fit into a block is form 1 to what ever will fit in the block specified. As an example for you a single 3mm x 5mm Brass mount standard of Almandine Garnet will cost £65. Delivery will be 1 week from receipt of your order.
} Rick Felten } 12/02/97 03:06 PM } I am looking for some Pyrope Garnet, Mg3Al2Si3O12, to use as a X-Ray } Microanalysis Standard. I have tried many of the EM suppliers (Energy Beam } Sciences, Polysciences, Fullam, and Ted Pella) w/o any luck. I would be } amenable to another standard that has major levels of Mg, Si and O and a } minor level of Al and no other elements. Any suggestions?
Rick: If you just need a specimen of pyrope garnet and do not want to buy an expensive standards collection of several dozen standards, then I suggest you try one of the many Internet Rock Shops. A quick check at one: http://www.gemhut.com revealed that they have many gem quality specimens of pyrope garnet starting at $9.95. I've purchased several isolated mineral specimens like this for standards and they work very well. Stanley L. Flegler, Assistant Director Center for Electron Optics Michigan State University
At 10:10 AM 12/1/97 -0800, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I don't know if I can help but I haven't seen anyone suggest what we do. Rather than picking up the sections with the grids we traditionally pick them up with a loop before placing them on the grid. In short, we make a round loop by hand - about 3 mm in diameter although the size or shape can be dependent on your specimen - with wire intended for metal evaporation such that if lowered onto a water surface it picks up the water droplet inside the circumference. This loop is then attached to the end of an applicator stick. THe loop is lowered onto the water surface around the sections - the sections are picked up with the water droplet, and then the entire contents are lowered onto a grid which is sitting on filter paper. With a film coating on the grid you might want to have your loop slightly larger than the grid so that the water flows out around it into the fitler paper - or cut "V" shaped pieces of filter paper to hold around the loop as you lower it onto the grid. I don't know if this will solve your problem but it's cheap and might be worth a try. Good luck!
Pat Hales McGill University Dept. of Anatomy & Cell Biology hales-at-hippo.medcor.mcgill.ca
We are interested in getting a digital camera for light microscopy. The simpler, the better. Just want to capture a good quaility image into a PC or SGI O2 platform that could then be handled by Photoshop or the like. We are considering both color and B7W only cameras. Cost is a consideration, as usual.
I would appreciate opinions from users and information from dealers.
TIA Greg Erdos
******************************************************* G.W. Erdos, Ph.D. Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab PO Box 118525 Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/
***** "Many shall run to and fro, and knowledge shall be increased" Daniel 12:4
Dear All, Does anyone have a better idea to detect intracellular calcium than by using the potassium pyroantimonate method as suggested in Hayat? We've tried it but we're not sure if it's precipitating calcium or the cesium that we use to induce metamorphosis in our experimental animals. Our thesis depends on it. Help!
-------------------------------------------------------- Name: Winston W Wiggins, Supervisor Vox:704/355-1267 CRC-Electron Microscopy Lab Fax:704/355-7648 Carolinas Medical Center Lab:704/355-7220 P.O. Box 32861 Charlotte, NC 28232-2861 USA Date: 12/3/97 E-mail: wwiggins-at-carolinas.org Time: 11:59:18 AM --------------------------------------------------------
a researcher wants me to prepare some thin section micrographs of red and white blood cells. Simply adding 2X fixative to a few hundred microliters of blood resulted, as expected, in a large clot.
Could someone recommend a simple protocol for such a sample--my botanical biases are showing through
thanks is advance
steve
--------------------------------------------------------------------- Dr. Steven Barlow, Associate Director EM Facility/Biology Department 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/
We have been using the Pixera system for a little more than a year now. It does color or B/W up to 1200 pixels across. The original cost was around $1200 for camera and interface, I think.
We started with it on a 486-66 and it was a bit slow. We now have it running on a 200 MHz Pentium thru a PCI card and it works okay for a inexpensive solution. The preview is a little slow for focusing but workable. The camera is TWAIN-compliant, sort of. The preview images to other imaging apps have the colors goofed up as far as our version of the software (1.15, I think), but the stored images are ok.
We ordered a 0.5x relay lens from Edmund Scientific for about $230 to adapt the camera to our photo-tubes. We lose some of our field of view (compared to our Polaroid camera) since the Pixera uses a 1/3" CCD, but it is adequate.
At 11:41 AM 12/3/97 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We also needed a digital camera with good quality image and versatile. After testing many we bought the PHotometrics Sensys. It is monochrome,chilled (10 degrees) 1400 chip. We have been very pleased with the resolution and sensitivity.
Bob
On Wed, 3 Dec 1997, Greg Erdos wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } We are interested in getting a digital camera for light microscopy. The } simpler, the better. Just want to capture a good quaility image into a PC } or SGI O2 platform that could then be handled by Photoshop or the like. We } are considering both color and B7W only cameras. Cost is a consideration, } as usual. } } I would appreciate opinions from users and information from dealers. } } TIA Greg Erdos } } ******************************************************* } G.W. Erdos, Ph.D. Phone: 352-392-1295 } Scientific Director, } ICBR Electron Microscopy Core Lab } PO Box 118525 Fax: 352-846-0251 } University of Florida E-mail: gwe-at-biotech.ufl.edu } Gainesville, FL 32611 http://www.biotech.ufl.edu/~emcl/ } } ***** } "Many shall run to and fro, and knowledge shall be increased" } Daniel 12:4 } }
Any idea is better than potassium pyroantimonate. Find the nearest friendly electron probe analyst (Peter Ingram or Ann LaFurgey?) and see whether they can help you, but it depends on Ca concentration that you seek. Good luck!
On Wed, 3 Dec 1997 wwiggins-at-carolinas.org wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear All, } Does anyone have a better idea to detect intracellular calcium } than by using the potassium pyroantimonate method as suggested } in Hayat? We've tried it but we're not sure if it's precipitating } calcium or the cesium that we use to induce metamorphosis in our } experimental animals. Our thesis depends on it. Help! } } -------------------------------------------------------- } Name: Winston W Wiggins, Supervisor Vox:704/355-1267 } CRC-Electron Microscopy Lab Fax:704/355-7648 } Carolinas Medical Center Lab:704/355-7220 } P.O. Box 32861 } Charlotte, NC 28232-2861 USA Date: 12/3/97 } E-mail: wwiggins-at-carolinas.org Time: 11:59:18 AM } -------------------------------------------------------- } }
I, and others in my situation, would appreciate hearing of contract labs that perform thin sectioning and other TEM related services. It would be helpful to hear testimonials of satisfaction (or otherwise) as well as names, telephone numbers, turnaround times, pros and cons. TIA.
Steve
Steven Samuelsson, Ph.D. Procter & Gamble Pharmaceuticals, Inc. PO Box 8006 Mason, OH. 45040-8006 (513) 622-1753 office (513) 622-1752 lab (513) 622-1196 fax samuelsson.sj-at-pg.com
You may wish to use scanning probe microscopy as it allows some friction, elasticity and conductivity data as well as molecular imaging at 37 C in bio buffers.
Zhifeng Shao of Virginia is doing some work on muscle, but there is a big difference between muscles in different parts of the body.
George
} From: Katri Vuopala {katri.vuopala-at-juniper.pp.fi} } To: "'Microscopy-at-MSA.Microscopy.Com'" {Microscopy-at-sparc5.microscopy.com} } Subject: how much is too much lipid in muscle-em? } Date: Tue, 2 Dec 1997 21:26:17 +0200
You might try high resolution in situ SPM as it can give you friction, elasticity, and conductivity data while doing molecular imaging at 37 C in biological buffers. The ease of use (simplicity of sample prep) is often a time saver over SEM.
Zhifeng Shao of U Virginia works on muscle, but there is a big difference between muscles in different parts of the body.
George
} } From: Katri Vuopala {katri.vuopala-at-juniper.pp.fi} } } Subject: how much is too much lipid in muscle-em? } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
____________________________________________________________________ ____________________________________________________________________ George Sibbald, President Molecular Imaging Corporation; Technology leader "in situ" SPM 9830A South 51st Street, Suite A124 Phoenix, AZ 85044, USA Phone(602)753-4311, Fax(602)753-4312 http://www.molec.com/
} Dear All, } Does anyone have a better idea to detect intracellular calcium } than by using the potassium pyroantimonate method as suggested } in Hayat? We've tried it but we're not sure if it's precipitating } calcium or the cesium that we use to induce metamorphosis in our } experimental animals. Our thesis depends on it. Help! } } -------------------------------------------------------- } Name: Winston W Wiggins, Supervisor Vox:704/355-1267 } CRC-Electron Microscopy Lab Fax:704/355-7648 } Carolinas Medical Center Lab:704/355-7220 } P.O. Box 32861 } Charlotte, NC 28232-2861 USA Date: 12/3/97 } E-mail: wwiggins-at-carolinas.org Time: 11:59:18 AM } -------------------------------------------------------- } Dear Winston,
I recommend the following methods:
1. Bichromate - Probst W., Histochemistry 85, 231-239, 1986.
2. Fluoride I - Ponie and Epel, J Histochem and Cytochem, vol 35,no 9, 939-956, 1987.
3. Fluoride II - Vohringer P., Microscopy Res and Technique, vol 31, 317-325, 1995.
I have been using the bichromate method mostly and with great success. Antimonate is very close in x-ray energy to calcium so it is difficult to support the findings of precipitated calcium by using EDX. This is not a problem with bichromate.
Good luck!
=============================================================== Rune Sundset Dept. of Medical Physiology, Inst. of Medical Biology, University of Tromsoe, N-9037 Tromsoe Phone : +47 77 67 54 42 or +47 77 64 46 96 Fax : +47 77 64 54 40 http://www-users.fm.uit.no/~knutst/medfys/medfys.htm --------------------------------------------------------------- Private Tunveien 21, D-10, N-9018 Tromso,Norway Phone: +47 77 67 45 48 ===============================================================
Hello I am looking for the distributor of AUROBEADS, colloidal gold used for coupling to proteins. It is not longer available from Amersham, but they couldnt tell were to get it now. reinhard
I wanted to find out about the pros and cons of leasing an SEM. I would also like to hear from any vendors off line, who could provide me with some information as well.
Thanks,
Paula
Paula Allan-Wojtas Atlantic Food and Horticulture Research Station Kentville, Nova Scotia B4N 1J5 Canada
I would appreciate if anyone could let me know of companies that do phosphor screen restoration. Please e-mail me directly at my address. Thank you in advance. Cathy Kelloes
I would appreciate if anyone could let me know of companies that do phosphor screen restoration. Please e-mail me directly at my address. Thank you in advance. Cathy Kelloes
We have been using carbon replicas of proteins sprayed or adsorbed on mica. It used to work quite well, but recently we have had problems with floating replicas off the mica support. It is the same batch of mica as before and I do not think we changed anything. Any bright ideas?
Thanks,
Michal
Michal Jarnik Lab of Structural Biology Research, NIAMS, National Institutes of Health, Bethesda, Maryland 20892. Ph: (301) 435-2587
I posted a message yesterday about the ASU Workshop. Apparently some people had problems reading the web page. I apologize. If you want information about the ASU Workshop on In-Situ Electron Microscopy and the ASU Winter School in January 1998, please see the following website: http://www.asu.edu/clas/csss/
John C. Wheatley Lab Manager Arizona State University Center for Solid State Science PSA-213 BOX 871704 Tempe, AZ 85287-1704
Our Sorval MT 2B needs an overhaul. Are there any who you would suggest in the proximity of Boston who can service an MT 2B? Thank you. Thane Benson {thane-at-epl.meei.harvard.edu}
We are in the market for a digital image acquisition system to retrofit to our 100CX. Could you suggest vendors who supply such a system? Thank You Thane Benson {thane-at-epl.meei.harvard.edu}
I am looking for sources of LR White embedded human muscle tissue suitable for IEM. The ideal situation would be if someone has, or could prepare, unstained sections on grids. Alternately, if I could locate LR White embedded material I could section it and return the block. Any help would be greatly appreciated.
Brett M. Connolly, Ph.D. Merck Research Laboratories Human Genetics Dept. WP26A-3000 PO Box 4 West Point PA 19486
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I would be interested in your findings. Elaine
Dr. Elaine Humphrey Biosciences Electron Microscopy Facility University of British Columbia 6270 University Blvd Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-unixg.ubc.ca
I too have been looking at low cost options for video/photomicroscopy. = I was ready to go for the color ccd and a Snappy. Then my advisor said = he would consider purchasing a Digital camera of moderate cost. The main problem with the mid-range digitals, as I am sure many of you = are aware, is that they are all fixed lense cameras. I noticed the Kodak = DC120 ($800) because it has fairly high resolution, it has threading on = the front of the lens to accept filters and adaptor lenses, and it also = has a macro mode. When I spoke with their tech. service, they told me = that a photomicroscopy system utilizing the DC120 system should be = released in late December (approximate cost - $2100). The system will = include the camera to C-mount adaptor, cables, power supply and software = for onscreen preview of images. For my application, a nice thing about = this system is that it uses the stock camera. The camera can then be = used for other tasks around the lab.=20 I actually bought the camera on my own to give it a try before trying = to sell the idea to my advisor. Of course, I didn't have any of the = adaptors or the luxury of the onscreen image. The camera easily = balanced atop a widefield ocular lens in the phototube. I was able to = preview pictures in the DC120's lcd display. With a litte trial and = error adjustment of the cameras lens position in macro mode I was able = to achieve simultaneous focus through the binocular and the camera.=20 With this crude system, I was able to get great quality images of = Acanthamoeba under phase contrast. I was most interested and concerned = with the cameras ability to capture fluorescent samples. I did get good = images of AO stains in the range of 1 to 8 second exposure times.=20 With very litte fuss, I was also able to take good quality gel photos = with the camera balanced on top of the polaroid gel hood.=20 My impression was that if you machined your own adaptor, the system = purchase might not be necessary but it would certainly be more = convenient.=20 My advisor was sufficiently impressed, but the verdict is still out on = whether I'll get to order the system.
Kevin Brent Smith - Masters Student University of Louisville Biology Dept. Life Science Bldg. Rm.#12 Louisville, KY 40292 Phone: (502) 852-6773 Fax: (502) 852-0725
We are research microscopists at Polaroid who have been using the Polaroid DMC Digital Microscope Camera in our laboratory since before its introduction in July of this year. We are very pleased with the high resolution digital images acquired with this camera. The DMC is an affordable (just under $6K) high resolution color CCD camera for the light microscope. The million pixel 3/4" CCD camera has a built in C-mount interface, which takes standard 1" or most 2/3" C-mounts. No special adapters are required. It is a SCSI device (no frame grabber board required) and quickly transfers images to your computer without any compression involved. It comes with a TWAIN driver, Adobe Photoshop plug-in for the Mac, and also a stand alone program (PC and Mac) to acquire and perform some image processing on images. The camera captures 24 bit color images in two sizes: a 1600 X 1200 pixel 5.5 MB image, or an 800 X 600 pixel 1.4 MB image. There is also an 8 bit B/W capture mode. The camera driver has a B/W preview mode with up to 5 frames/second capture to aid in positioning and focusing. A focus meter mathematically monitors and helps you optimize focus. Please visit Polaroid's website for more information - www.Polaroid.com.
At 11:15 AM 12/4/97 -0500, Thane Benson wrote: } Our Sorval MT 2B needs an overhaul. Are there any who you would suggest } in the proximity of Boston who can service an MT 2B?
I would recommend Bill McGee. He worked for Dupont-Sorvall when they made this microtome. His company is Microtome Service Company of Liverpool, NY. He can be reached at 315-451-1404.
Best regards, Steven E. Slap ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
It really depends on what you want to do with the images i.e. print them = out to photo quality printer, etc. We have a 100CX with an image system on it, but could not afford the high = end system. It is likely that this will prevent us from truly using it = for what it was intended although the manufacturer indicated that it = should do the job i.e. print most of our images directly to a Codonics = NP1660 printer. What will you be using it for?
Judy Murphy Microscopy Technology Center San Joaquin Delta College 5151 Pacific Ave. Stockton, CA 95207 209/954-5284 __________________________________________________________________________= _____
We are in the market for a digital image acquisition system to retrofit to our 100CX. Could you suggest vendors who supply such a system? Thank You Thane Benson {thane-at-epl.meei.harvard.edu}
------------------ RFC822 Header Follows ------------------ Received: by sjdccd.cc.ca.us with ADMIN;4 Dec 1997 11:15:50 -0800 Received: from Sparc5.Microscopy.Com (206.69.208.10) by = ms.sjdccd.cc.ca.us with SMTP (Eudora Internet Mail Server 1.2); Thu, 4 Dec 1997 11:15:26 = -0800 Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com = (8.6.11/8.6.11) id KAA14682 for dist-Microscopy; Thu, 4 Dec 1997 10:05:09 = -0600 Received: from epl.meei.harvard.edu (epl.meei.harvard.edu = [204.166.79.51]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id = KAA14679 for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 4 Dec 1997 10:05:08 = -0600 Received: from 204.166.79.204 (amb-office.meei.harvard.edu) by = epl.meei.harvard.edu (M/EPL-97.11.01) id AA04939; Thu, 4 Dec 97 11:19:14 EST Message-Id: {3486D802.EB6-at-epl.meei.harvard.edu}
Hello All,
Does anyone know what has happened to Dr. Om Johari and the journal
{bold} Scanning Microscopy {/bold} . I have been unable to contact him via e-mail, phone,
or fax and have not been able to find reference to the journal in over two
A colleague has a dust problem. He has several mobile carts used to transport plastic components that he wishes to keep free of dust and very clean. The components are kept inside of a 36 cubic inch box on top of the cart. Anyone know of a way to keep the components from attracting dust when the box is opened to remove one of the plastic parts?
Since the carts are going to be moved a lot, the anti-static device should also be mobile. I suggested electrostatic precipitators, but he did not like the idea of high voltages? Likewise, he did not like my idea of using an alpha emitter (Americium, for example). Can't think of much else, however.
Thanks.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing 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 ####################################################################
Gaylord Garroway has rendered routine maintenance service for our Sorvall microtomes (we have 9) for over 20 years. He is very competent, and we have been pleased with his service. He may be reached at 1-508-473-9579 (Milford, MA).
Vachik Hacopian
} Our Sorval MT 2B needs an overhaul. Are there any who you would suggest } in the proximity of Boston who can service an MT 2B? } Thank you. } Thane Benson {thane-at-epl.meei.harvard.edu}
We're setting up a new lab and we will be doing a whole lot of microscopy: light, confocal, EM. We are looking for recommendations on a versatile printer with which we can print publication-quality (at least 600dpi but preferably better) prints. Anyone willing to tell all about their printer? -=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-= Paul Tiseo | "It's funny that pirates were always going Mayo Clinic - Jacksonville | around searching for treasure, when they Birdsall 3 | never realized that the real treasure (904) 953-8254 (pager) | was the fond memories they were creating." tiseo.paul-at-mayo.edu | http:// coming soon | - Jack Handey -=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=
No reason why you can't embed the clot to see RBCs, but you won't have many WBCs. To see lots of these, you need a buffy coat. You can either separate them with Lymphocyte Separation Media or Ficoll Hypaque, or you can just let the cells settle out (heparinized) on the benchtop or in a low speed centrifuge. Gently remove the serum and gently add glut without disturbing the pellet. Let fix for a couple of hours and take off the very top layer (either cut the plastic tube with a razor blade, or remove the cells with a Pasteur pipet. This layer will be enriched with WBCs, but will also have many reds. It will appear slightly pinker than the cells in the bottom (a creamy hue). Re-pellet and encase with agar to keep them together as a block while embedding.
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
It seems to me that you are forced to go ex situ with SEM?
It would seem that you will loose versatility rather than gain. The current SPM design can give you atomic resolution with STM and AFM, Plus allow control of the environments (PH, Temperature, Electro chem Potential) and work under liquids (wide range of viscosity).
For food studies you get the atomic resolution imaging while you simulate real conditions such as digestion, cooking, freezing, reactions with drugs, alcohol, etc.
This is also the big interest from Biological SEM users moving to SPM for in situ studies on live samples at 37 C.
To my knowledge what may be missing is spectroscopy.
But you gain dramatically by moving to high resolution in situ microscopy.
Let me know about your needs, SPM may be a good alternative.
George
____________________________________________________________________ ____________________________________________________________________ George Sibbald, President Molecular Imaging Corporation; Technology leader "in situ" SPM 9830A South 51st Street, Suite A124 Phoenix, AZ 85044, USA Phone(602)753-4311, Fax(602)753-4312 http://www.molec.com/
Dr. Elaine Humphrey wrote: =================================================== I wanted to find out about the pros and cons of leasing an SEM. I would also like to hear from any vendors off line, who could provide me with some information as well. =================================================== As some one who has purchased outright SEMs, bank financed SEMs and leased SEMs over the past almost thirty year period, think I can answer with some amount of experience. And the answer: It is "all in the eyes of the beholder", or putting it another way, it is all a matter of trade offs between interest rates, bank loan rates vs. lease rates. The decision that is the right one at one point in time, might be the wrong decision at some other time.
And this kind of decision might be one way in the USA and it might be an entirely different one in Canada where there are different tax laws and different rules relating to the tax deductibility of lease payments as a business expense.
In general, the "best deal" is to pay cash but you also have to take into consideration the return you would other wise be getting if you left that money invested where it was and you took out a bank loan to pay for the instrument. In other worlds, it all depends on interest rates at the time the decision has to be made.
Leasing, is in general, just a more expensive form of bank financing. It is more expensive because the leasing company has to worry about what they would do if they had to foreclose, say, on someone's TEM. This risk can be partially off-set by a buy-back guarantee from the manufacturer, but it has been our own experience that a manufacturer, since they now have to cover this added "cost" or "risk" of having to undo the sale some day, tends to be a bit less competitive on the final negotiated selling price.
Even the decision whether to use an "open" or "closed ended" lease depends on who is going to carry the burden of risk of predicting fair market value at the end of the lease term.
So the real answer is that there are a number of trade offs, the sum total of which are influenced by a) your country, b) your tax status (e.g. for profit vs. nonprofit vs. government lab), and c) relative lease vs. bank loan rates. This has, for us, always been an exercise for our outside accounting firm and not for us scientists or even our in-house accounting department people.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Does anyone have any experience with analysis of particulates on Nucleopore filters? We have an ongoing project involving particle counts and EDS analysis of air sample particulates collected on Nucleopore filters. For particle counts, I have been cutting out sections of the filters and attaching them with carbon tape to aluminum stubs, then gold coating them. For EDS, I have simply attached cut sections of the filters to clean carbon stubs with carbon tape and viewed/analyzed them using the variable-pressure mode of our SEM. Since we need to image particles in detail at mags up to 20,000x, it's not feasible to do both imaging and EDS on uncoated samples, due to resolution limitations of backscatter imaging and variable pressure conditions.
On occasion, it has seemed that fewer particles are seen on the sputter-coated samples than on the uncoated ones, although they are taken from the same filters from adjacent locations. Since the particles themselves are only attached loosely to the collecting filters (i.e., no special adhesive techniques are used), is it possible that the sputter-coating process can dislodge significant numbers of particles? Needless to say, this could have serious consequences for the data....
Thanks for any feedback on this.
Randy Tindall 2017 Princess Jeanne Las Cruces, New Mexico 88001-4157
How about slight positive pressure while the cabinet is opened? Pressure could be provided from a small compressed air or nitrogen cylinder. This should ensure that no particles from outside the cabinet would enter - if the opening is not too large. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au
} } A colleague has a dust problem. He has several mobile carts used to } transport plastic components that he wishes to keep free of dust and very } clean. The components are kept inside of a 36 cubic inch box on top of the } cart. Anyone know of a way to keep the components from attracting dust } when the box is opened to remove one of the plastic parts? } } Since the carts are going to be moved a lot, the anti-static device should } also be mobile. I suggested electrostatic precipitators, but he did not } like the idea of high voltages? Likewise, he did not like my idea of using } an alpha emitter (Americium, for example). Can't think of much else, } however. } } Thanks. } } } #################################################################### } John J. Bozzola, Ph.D., Director } Center for Electron Microscopy } Neckers Building, Room 146 - B Wing } Southern Illinois University } Carbondale, IL 62901 } U.S.A. } Phone: 618-453-3730 } Fax: 618-453-2665 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/departments/shops/cem.html } #################################################################### } }
This was my thought also, but keep in mind that the gas in cylinders is not always "particle free" either. I would suggest that you filter the gas just as it enters the cabinet. Rating of the filter should be determined by how small a particle you are concerned with. My bias is that I am a microscopist for a filter company, but I am continually suprised at how dirty many "new" materials are as delivered.
Bob Holthausen Pall Corporation Port Washington, NY
jim-at-proscitech.com.au on 12/05/97 01:11:40 AM
To: bozzola-at-siu.edu cc: microscopy-at-Sparc5.Microscopy.Com (bcc: Bob Holthausen/SLSNY/Pall/US)
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
i do similar analyses in some undergraduate courses at the UofR. the technique we use is to collect on 0.2um pore size 13mm filters. after the particles are collected i take a 1/2" pin mount and coat it with carbon paint. while the paint is still wet i put the whole filter in it...it sticks well and the carbon does not migrate up through the pores (let it air dry in a clean area though). i then coat with either Au or C depending on the intention of EDS work...
i suspect that you may be losing particles in handling; the technique above may avoid some of these concerns (although small particles are pretty intimately attached to the polycarbonate filters)
hope this helps
b-
**************************************************************** Brian McIntyre Electron Microscopy Lab Institute of Optics University of Rochester Rochester, NY 14627
} The main problem with the mid-range digitals, as I am sure many of you are aware, } is that they are all fixed lense cameras.
That is a problem but not the main one. The main one is that most "consumer level" cameras do compression onthe image - usually JPEG - to reduce file size. This is absolutely inimical to subsequently trying to do any serious analysis on the images later - details are altered, moved, etc., brightness and color altered differently in different regions, etc. The "serious" cameras like the Kodak DCS and Polaroid DMC (I use the latter) ship the image to the computer without compression. John Russ
I HAVE A COMMERCIAL STAKE IN WHAT I AM ABOUT TO TELL YOU!
Have you seen/tried the Polaroid DMC? It connects to the microscope via standard C-mount (no lens on the camera, just C-mount thread), creates a TIFF file into your computer at 1600x1200 or 800x600 pixel resolution, and converts quickly and easily for macro work on your copy stand by adding C-mount macro lens. List price under $6K. Details on the Polaroid website at http:\\www.polaroid.com
Hoping this isn't too commercial. The product is still rather new (introduced July '97) and seems directly applicable to your question.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I too have been looking at low cost options for video/photomicroscopy. I was ready to go for the color ccd and a Snappy. Then my advisor said he would consider purchasing a Digital camera of moderate cost. The main problem with the mid-range digitals, as I am sure many of you are aware, is that they are all fixed lense cameras. I noticed the Kodak DC120 ($800) because it has fairly high resolution, it has threading on the front of the lens to accept filters and adaptor lenses, and it also has a macro mode. When I spoke with their tech. service, they told me that a photomicroscopy system utilizing the DC120 system should be released in late December (approximate cost - $2100). The system will include the camera to C-mount adaptor, cables, power supply and software for onscreen preview of images. For my application, a nice thing about this system is that it uses the stock camera. The camera can then be used for other tasks around the lab. I actually bought the camera on my own to give it a try before trying to sell the idea to my advisor. Of course, I didn't have any of the adaptors or the luxury of the onscreen image. The camera easily balanced atop a widefield ocular lens in the phototube. I was able to preview pictures in the DC120's lcd display. With a litte trial and error adjustment of the cameras lens position in macro mode I was able to achieve simultaneous focus through the binocular and the camera. With this crude system, I was able to get great quality images of Acanthamoeba under phase contrast. I was most interested and concerned with the cameras ability to capture fluorescent samples. I did get good images of AO stains in the range of 1 to 8 second exposure times. With very litte fuss, I was also able to take good quality gel photos with the camera balanced on top of the polaroid gel hood. My impression was that if you machined your own adaptor, the system purchase might not be necessary but it would certainly be more convenient. My advisor was sufficiently impressed, but the verdict is still out on whether I'll get to order the system.
Kevin Brent Smith - Masters Student University of Louisville Biology Dept. Life Science Bldg. Rm.#12 Louisville, KY 40292 Phone: (502) 852-6773 Fax: (502) 852-0725
Paul, I would recommend looking into a Tektronix 450? color wax printer. We have an older 350 on our network that is great for color and greyscale. I believe ours is 300 dpi. 600 dpi files will take up an enourmous amount of memory. Journals will trash your 600 dpi images back to 300 dpi anyway when reducing to half-tone. Only drawback is you cannot write directly on the images Photoshop annotation as a seperate layer works well). Just my .02 worth.
ed
Edward J. Basgall, PhD The Pennsylvania State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618 http://www.personal.psu.edu/ejb11/ Privilege does not absolve one of ecological responsibility.
At 10:20 PM 12/4/97 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I do SEM/EDX particle analysis on a fairly regular basis. I had also found that samples which I had sputter coated had fewer particles. I now use just sticky carbon tabs for most of my particles, but the ones in liquids I first pick up on a grid, and then mount on the sticky carbon. These can be sputter coated without losing the particles. If you absolutely have to collect your particles on a Nucleopore, you might try collecting as usual, and then using a conductive tape/tab to pick up the particles from the Nucleopore, then mounting the conductive tape/tab.
Stephanie Wind McCray Process Chemist Moltech Corp. 9000 S Rita Rd, Bldg 61 Tucson, AZ 85747 520-799-7631 (office) or 520-799-7535 (lab) wind-at-moltech.com
} Hi, } } Does anyone have any experience with analysis of particulates on Nucleopore } filters? We have an ongoing project involving particle counts and EDS } analysis of air sample particulates collected on Nucleopore filters. For } particle counts, I have been cutting out sections of the filters and } attaching them with carbon tape to aluminum stubs, then gold coating them. } For EDS, I have simply attached cut sections of the filters to clean carbon } stubs with carbon tape and viewed/analyzed them using the variable-pressure } mode of our SEM. Since we need to image particles in detail at mags up to } 20,000x, it's not feasible to do both imaging and EDS on uncoated samples, } due to resolution limitations of backscatter imaging and variable pressure } conditions. } } On occasion, it has seemed that fewer particles are seen on the } sputter-coated samples than on the uncoated ones, although they are taken } from the same filters from adjacent locations. Since the particles } themselves are only attached loosely to the collecting filters (i.e., no } special adhesive techniques are used), is it possible that the } sputter-coating process can dislodge significant numbers of particles? } Needless to say, this could have serious consequences for the data.... } } Thanks for any feedback on this. } } Randy Tindall } 2017 Princess Jeanne } Las Cruces, New Mexico 88001-4157
Hi Randy, A trick I have had some success with is to pre-coat the filters with a conducting metal, Au or AuPd before collecting particulates. If there is peak overlap you might even try a Cr replicate. It does a great job of making polycarbonate filters conductive, I also attach them with Ag paint. Since we're using a mass spec technique (TOF-SIMS) the carbon tape adhesive shows up. Another advantage to the AuPd pre coating is that I can use the peaks as a SIMS calibration aid.
I have used this to investigate uncoated, unfixed, freeze dried yeast cells with both LVFESEM (2kV) and TOF-SIMS. I can't say that I've tried EDS on them. The SEMs come out great up to about 9000x. I've posted some images on my web-site, follow the links to "Research Projects" then to "Sample prep for LVSEM and TOF-SIMS".
good luck ed
Edward J. Basgall, PhD The Pennsylvania State University Surface Chemistry Group ejb11-at-psu.edu Materials Research Institute Building Ph: 814-865-0493 University Park, PA 16802-7003 FAX: 814-863-0618 http://www.personal.psu.edu/ejb11/ Privilege does not absolve one of ecological responsibility.
I' tryng to collect all info possible about "Image Plates" devices.
I know that FUJI had in the past a sytem called FDL 5000. (I have a 1 page leaflet). Fuji, in Italy does know nothing about this produc. Can anyone help me with more info, approx. price, where to find it, etc.
Regards to everybody - Have a nice Christmas and happy New Year !
PLEASE REMOVE "_NO_SPAM" and "NO_SPAM_" before and after -at- symbol to reply me.
We recently purchased the Epson Sylus Photo printer (only costs about $500) to use as an intermediate-level printer. However, we have been pleasantly surprized at the quality we get when using the Epson Photographic Quality Glossy *Film*. A colleague recently submitted such prints of histology samples for publication.
The printer resolution goes up to 720 dpi, and there are 6 color jets vs. the usual 4. It is a little slow (takes about 5-8 minutes to do high resolution full sized prints), but we don't have a large volume so that's OK. The only problem we've had is getting the color settings to produce consistent white backgrounds at the same time as producing reasonable representation of reality. This is an issue not only of printer settings but computer software, monitor, etc. However, even when the image on the monitor is reasonable, the printout can be quite different. Takes a bit of fiddling to get desired color representation.
The usual disclaimer: no connection to Epson other than customer.
Karen
tiseo.paul-at-mayo.edu wrote: } } Hi, } } We're setting up a new lab and we will be doing a whole lot of } microscopy: light, confocal, EM. We are looking for recommendations on a } versatile printer with which we can print publication-quality (at least } 600dpi but preferably better) prints. Anyone willing to tell all about their } printer? } -=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-= } Paul Tiseo | "It's funny that pirates were always going } Mayo Clinic - Jacksonville | around searching for treasure, when they } Birdsall 3 | never realized that the real treasure } (904) 953-8254 (pager) | was the fond memories they were creating." } tiseo.paul-at-mayo.edu | } http:// coming soon | - Jack Handey } -=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=
-- Karen Zaruba kszaruba-at-mmm.com 3M Company, St. Paul, MN 55144 "Opinions above are my own, not necessarily my employer's"
Microscopy members, I am finishing a paper on the "Evolution of microscopy in the XX century", (in hypertext) and have been unable to find pictures of Zworikin and Oatley in the locally available literature. If anyone can help me by attaching a .jpg, tif or similar picture to email, I would be very grateful.
please answer directly to
wamann2-at-metalmat.ufrj.br
Prof. Walter A. Mannheimer Dept. of Metallurgy and Materiais Eng. Federal University of Rio de Janeiro POBox 68505, 21945 Rio de Janeiro, Brazil Vox (55 21) 590-0579 Fax (55 21) 290-6626 wamann-at-metalmat.ufrj.br
} } Hi there we are looking for a very small objective aperture for our JEOL } 2000FX. Since these apertures come in strips there is a limited selection } and the "standard" set comes with a 20micorn aperture as the smallest. We } can get a 10micron from JEOL, does anyone know of a smaller one say } 5microns? } I have not called all of the Microscope Spares and Equipment suppliers yet, } I thought I would solicit commnets for the community. Many thanks. } Reply by email and I will summarize to the list if there is sufficient } interest and response. } } Cheers } } Jfm.
} Dear John,
Ladd has been producing Apertures for over forty years now. Since we produce them ourselves we can give you any hole size that you wish, within certain technical limitations. Please e-mail me with the size of all the holes you would like on the strip and the material, I suspect PT, and we will send you a quote.
Best Regards,
JD Arnott Ladd Research 13 Dorset Lane Williston, VT 05495
TEL 1-802-878-6711 worldwide 1-800-451-3406 US, Canada Fax 1-802-878-8074
Ann Fook Yang wrote: ================================================ I am planning to do immunogold on brassica pollen grains. I am looking for an adhesive that remain tacky when dried and would hold pollen through out the process of washing, fixing, immuno-treatment, postfixing, dehydrating and critical point drying.
Any suggestions will be appreciated. Thanks. ================================================ You might want to consider trying SPI's Tacky Dot(TM) Slides which can be found on our website (along with an example of their use) at URL
http://www.cccbi.chester.pa.us/spi/new/tacky.html
If the pollen is reasonably free flowing, one grain (or possibly several) will end up "sticking" per dot and since there is a build up of strength of the adhesive bond with time (as is the case for most adhesives), after about 48 hours it is at its maximum. For most particles, the bond is reasonably resistant to water, especially if not subjected to turbulent conditions. If you can do the preparation on the slides, the end result will be infinitely more easy to characterize with any kind of microscope than if the pollen grains are in some random distribution on a substrate.
The "adhesive" is a dry adhesive, there is no chance of any particle sinking into it, and there is nothing to off-gas to contaminate a vacuum system. One mounted, so long as the slides are stored long term under dry conditions as would any other SEM specimen, their life time seems to be indefinite.
Disclaimer: SPI Supplies manufactures Tacky Dot Slides under license from DuPont, the patent holder and we therefore would like to see more people using them! We are unaware of any other product of this type in the world.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
One of the cheapest and simplet "glues" for pollen grains is to dissolve the glue from about a 30cm length of Scotch tape in about 10ml of chloroform. Apply this to a clean abd shiny stub allow to d5ry and sprinkle on or place pollen grains on the surface. Dry6 overnight in a 35oC oven, loghtly coat 8-10nm Au/Pd and away you go.
Patrick Echlin Cambridge UK
On Sun, 30 Nov 1997, Garber, Charles A. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Ann Fook Yang wrote: } ================================================ } I am planning to do immunogold on brassica pollen grains. I am looking for } an adhesive that remain tacky when dried and would hold pollen through out } the process of washing, fixing, immuno-treatment, postfixing, dehydrating } and critical point drying. } } Any suggestions will be appreciated. Thanks. } ================================================ } You might want to consider trying SPI's Tacky Dot(TM) Slides which can be } found on our website (along with an example of their use) at URL } } http://www.cccbi.chester.pa.us/spi/new/tacky.html } } If the pollen is reasonably free flowing, one grain (or possibly several) } will end up "sticking" per dot and since there is a build up of strength of } the adhesive bond with time (as is the case for most adhesives), after about } 48 hours it is at its maximum. For most particles, the bond is reasonably } resistant to water, especially if not subjected to turbulent conditions. If } you can do the preparation on the slides, the end result will be infinitely } more easy to characterize with any kind of microscope than if the pollen } grains are in some random distribution on a substrate. } } The "adhesive" is a dry adhesive, there is no chance of any particle sinking } into it, and there is nothing to off-gas to contaminate a vacuum system. } One mounted, so long as the slides are stored long term under dry conditions } as would any other SEM specimen, their life time seems to be indefinite. } } Disclaimer: SPI Supplies manufactures Tacky Dot Slides under license from } DuPont, the patent holder and we therefore would like to see more people } using them! We are unaware of any other product of this type in the world. } } Chuck } } =================================================== } Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 } President 1-(800)-2424-SPI } SPI SUPPLIES FAX: 1-(610)-436-5755 } PO BOX 656 e-mail: cgarber-at-2spi.com } West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com } } } Look for us! } ############################ } WWW: http://www.2spi.com } ############################ } ================================================== }
I'm not usually one for "me too" posts, but all you people who are shelling out thousands of dollars for Textronics-level printers to make "publication quality" prints ought to heed Karen Zaruba's (and my) words. Today, the best bang-for-bucks is acheived with the Stylus Photo. If you find the Epson Glossy Film a bit pricey (at roughly $2.50/sheet, I do), you can go to Kodak Inkjet Photo Quality paper (it looks and feels like photo paper) for about one-third that cost and not lose much definition. The prints that come off the Epson device are publication quality - I defy anyone call them "unacceptable" especially given the previous post on what actually goes on in the transfer to print! I will submit some next week for publication and I plan not to say a word about how they were generated - let the printer complain (if they even notice). The only thing noticeably better than these prints is the digital image itself. Rob Palmer CEB/UT
Re the comment "Takes a bit of fiddling to get desired color representation." (Seems like I may have heard that once or twice before!)
One of the big benefits I hear for the Codonics printer is their "bracketing" feature, where it will print series of thumbnails of an image, varying several combinations of settings such as gamma, so that the best-appearing combination can be chosen easily without guessing. Does anyone know if such a program (maybe a RIP, Raster image processor program?) exists that can be used with other printers, e.g. Epson or HP? Any comments on price/benefit?
Thanks Richard Thrift DepoTech Corp. Richard_Thrift-at-DepoTech.com
} } } {kszaruba-at-MMM.COM} 12/05/97 09:40am } } } . . . We recently purchased the Epson Sylus Photo printer . . . . . . The only problem we've had is getting the color settings to produce consistent white backgrounds at the same time as producing reasonable representation of reality. This is an issue not only of printer settings but computer software, monitor, etc. However, even when the image on the monitor is reasonable, the printout can be quite different. Takes a bit of fiddling to get desired color representation.. . .
} Have you seen/tried the Polaroid DMC? It connects to the microscope } via standard C-mount (no lens on the camera, just C-mount thread), } creates a TIFF file into your computer at 1600x1200 or 800x600 pixel } resolution, and converts quickly and easily for macro work on your } copy stand by adding C-mount macro lens. List price under $6K. } Details on the Polaroid website at http:\\www.polaroid.com
How does this differ from the PDC-2000/T which, I believe, is much cheaper?
Article posted on Lindsay's Lab at ASU web shows force data of 7 liquid molecular layers at the solid liquid interface, molecular orientation, and molecular deformation based on position.
This combined with in situ control of environments should be the enabling technology for molecular tribology studies.
For corrosion and tribology studies you can get atomic imaging, as well as several types of force data LFM (lateral force) MAC Force (direct vertical force), MAC Phase data, and MAC Phase with fractionalized tips (chemical force)
George
____________________________________________________________________ ____________________________________________________________________ George Sibbald, President Molecular Imaging Corporation; Technology leader "in situ" SPM 9830A South 51st Street, Suite A124 Phoenix, AZ 85044, USA Phone(602)753-4311, Fax(602)753-4312 http://www.molec.com/
Will you be at Cell Biology show in Washington D.C., December 15 - 18?
If you are please come to talk to us about Scanning Probe Microscopy. We will be conducting training and "live" demonstrations of a technical breakthrough for in situ high resolution biological imaging.
George
At 11:02 AM 12/4/97 -0600, henk-at-vt8200.vetmed.lsu.edu wrote:
} } } }
{excerpt} ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello All,
Does anyone know what has happened to Dr. Om Johari and the journal
{bold} Scanning Microscopy {/bold} . I have been unable to contact him via e-mail, phone,
or fax and have not been able to find reference to the journal in over two
years. Have he and the journal "retired"?
Bill Henk
Dept. of Anatomy & Cell Biology
LSU School of Veterinary Medicine
Baton Rouge, LA 70803
phone -(504)346-3237
e-mail - henk-at-vt8200.vetmed.lsu.edu
{/excerpt} { { { { { { { {
F.Y.I. ASU/MI Winter Microscopy Workshop is hosting its annual AFM in Biology Hands-on Workshop. It will be held February 9th to 11th, 1998. This is a wonderful opportunity for someone to learn the fundamentals of AFM. On the second day of the workshop, participants will get a chance to image atoms and molecules. Participants are encouraged to bring their own samples for free testing.
I have used Me salicylate to clear whole (gutted, 100mm sculpins) fish. This was a pretty simple procedure, just dehydrate the fish in an ethanol series, then transfer to 100% Me salicylate through a 3:1, 1:1, 1:3 EtOH:Me sal. series. If parts stay cloudy, it is probably because they weren't completely dehydrated. I used 1 to 3 days in each step, tissue pieces would take less. When finished, the fish almost looked like they were made of glass.
Phil
} From: Ramin Rahbari 313 998-3383 {rahbarr-at-aa.wl.com} } } I am interested to hear from individuals that have used methyl salicylate (? } conc.) to clarify tissue in particular skin. }
Philip Oshel PO Box 5037 Station A Champaign, IL 61825-5037 (217) 355-1143 oshel-at-ux1.cso.uiuc.edu or poshel-at-hotmail.com ***** looking for a job *****
Disclaimer: I do represent a business that sells computers and their parts. We build and repair computers.
Where did you get the price figures for the hard drives? If you are talking about IDE, you are several years behind in prices. 6.0 Gig HDD's only retail for about $300.00 or so. Look around. The price of things may surprise you.
SCSI HDD's are sinking in price as of early summer. The price is very affordable.
The best way to back up is still via a second HDD. It is less expensive than most alturnatives and there is no media to concern yourself with. Take care of it and keep it clean and it should last for a long time.
Don't forget that we are on the horizon of 1.0 Gig floppy drives.
Things are getting better and better as well as less expensive.
____________________________________________________________________ ____________________________________________________________________ George Sibbald, President Molecular Imaging Corporation; Technology leader "in situ" SPM 9830A South 51st Street, Suite A124 Phoenix, AZ 85044, USA Phone(602)753-4311, Fax(602)753-4312 http://www.molec.com/
Hello All I am sorry - the message with above subject was dedicated to Microprobe list. regards Krzysztof M.Herman LabSoft Sp.C. 21 Kosciuszki Str. 05-500 Piaseczno, Poland tel/fax: (48 22) 7502024, 7502028, 7570671 fax: (48 22) 483787, mobile (48 90) 213438 E-mail: labsoft-at-labsoft.com.pl http://www.labsoft.com.pl/ ------=_NextPart_000_01BCFD25.4B37D9E0 Content-Type: text/html; charset=ISO-8859-2 Content-Transfer-Encoding: base64
If you be at Cell Biology show in Washington D.C., December 15 - 18, please come to talk to us (booth # 253) about Scanning Probe Microscopy.
We will be conducting training and "live" demonstrations of a technical breakthrough for in situ high resolution biological imaging.
George
F.Y.I. ASU/MI Winter Microscopy Workshop is hosting its annual AFM in Biology Hands-on Workshop. It will be held February 9th to 11th, 1998. This is a wonderful opportunity for someone to learn the fundamentals of AFM. On the second day of the workshop, participants will get a chance to image atoms and molecules. Participants are encouraged to bring their own samples for free testing.
____________________________________________________________________ ____________________________________________________________________ George Sibbald, President Molecular Imaging Corporation; Technology leader "in situ" SPM 9830A South 51st Street, Suite A124 Phoenix, AZ 85044, USA Phone(602)753-4311, Fax(602)753-4312 http://www.molec.com/
Paul, Don=92t like using their name so let=92s say Evex & similar=3D the gener= al term "Slime-x". I suspect that their product line consist of a broad, very broad but shallow array of airware, little else. No idea what their claims are. No reason to believe what I=92d see or hear. Don=92t care. I am glad to see this subject up again. During the intense frothing over the Slime-x ethics, it seemed the common denominator was frustration at the lack of punitive actions available. No cyber bullets....wrong!.... information is a wonderful thing. Well folks here is my shot. Our institution maintains a list of preferred suppliers as well as problem children. They make a reasonable effort to be fair in assessments. I supplied a sampling of the better researched messages, their unsigned responses & a memo suggesting that a company that behaved in this manor could not be trusted at any level. This is not an absolute fix but is progress. BANG I believe the we have an obligation to protect our counter parts in the field, most of which I dare to say do not subscribe to this list. For now, the low life ethical standards of Slime-x are an issue to us, probably to no one else. Many of our students are potential customers of high tech. hardware. Their decisions will reflect on & may impact us directly. Down the road, they/we may be oblivious to this companies ethical status. Any of you who have centralized purchasing or preferred vendor list can help many of our associates now & in the future by doing as I have done. Document the demon. Naturally there will be others who follow in the footsteps of Slim-x but I think a good public spanking is in order and a fairly effective deterrent to others.
from my extream side: in a loose paraphrase of George Patton in Hollywood... when you put your hand it the goo that used to be your funding's face you=92ll know how to kill the enemy, shoot them in the belly ....
Are there any Zeiss Wizards out there that can help me with a few questions concerning a Zeiss UNIVERSAL 'M' microscope ?
1-What are the electrical demands of a GLAREX viewing hood, and what does the electrical attachment do ?
2-What is the purpose of the light bulb on the 35mm camera attachment ?
PARTS WANTED:
1-Adaptor ring to attach light source for reflected light microscopy on back of microscope. ie: Adaptor ring 3.4 mm (internal thread) {-} 4.3 mm bayonet mount or Adaptor ring 3.4mm (internal thread) {-} 3.3 mm bayonet mount
2- Extension tubes (12 mm) for EPIPLAN lenses with M24mm thread
3- Low power (1 to 8x) EPIPLAN, EPIPLAN LD or EPIPLAN HD lenses
4- H-Pr-POL reflector for vertical Illuminator IIC and EPIPLAN POL lenses
ZEISS parts for sale/swap
1- Vertical Illuminator IIC (lens mount does not hold 'quick change ring') 2- Adaptor ring (brass, home made, 4.2mm thread (externior thread) {-} 4.3 mm bayonet mount.
OR, do you know the name, address, telephone number, e-mail of some one that does.
Hello!: I was wondering if one of your associates might help me. I have an older (20-30 years) microscope. It is a Spencer/American Optical, bifocal, oil immersion, serial number 155715, but with no other type number. The instruction manual was lost (prior owner) many years ago. Is there anyone out there who might have a copy of the manual that they might share with an ignorant biochemist? Please help. E Mail defilip1-at-flash.net. Thank you
I have found that floating carbon off mica, as with floating Formvar off glass slides, seems to be affected by a combination of the alignment of the planets, the phase of the moon and the day of the week :-)
Other factors appear to be: Humidity of the air in the room (if too dry, no good); Surface condition of the mica (I rinse in alcohol and acetone after I cleave the mica); Condition of the evaporation system (possible oil contamination etc.).
Mostly though I just wait a day or two and try again. Usually works.
JUST A QUICK LETTER TO SHOW YOU SOME LASERS- OPTICS AND OPTICAL TABLES SURPLUS THAT WE JUST RECEIVED.
ITEM TRIMMU12 14 WATT ARGON LASER MADE FOR HEART SURGERY, TRIMEDYNE MODEL 900 TEMOO, POLORIZED,220VAC INPUT , WATER COOLED , FIBER LAUNCH, ALL ON ROLLAROUND CART EXCELENT FOR LAB USE, THE POWER WAS MEASURED AT 13 TO 14 WATTS. PRICE $9500 12 MONTH WARRANTEE.
ITEM: COHERENT ARTICULATING ARM FROM A MODEL 451 CO2 MEDICAL LASER. ECCELLENT COND. $200
ITEM CO220A: CO2 LASER MADE BY PFIZER ,1990, FOR SURGERY, TATTOO REMOVAL ECT. 20 WATT OUTPUT , TESTED AND IN EXC. COND. 110 VAC INPUT, COST $40,000 NEW OUR PRICE 4,900. MODEL 20-C
ITEM:PDA-1U1 SPECTRA PHYSICS QUANTRA RAY PULSED DYE LASER , GOOD FOR SPARE PARTS MODEL PDA-1 $500
ITEM NEWU1 NEWPORT OPTICAL TABLE 16" BY 36" 4" THICK, 1 " HOLE SPACING, COMES WITH A RUBBER ISOLATED TABLE STAND, NOT AIR SUPPORTED, $750
ITEM: HEPSN1 HELIUM NEON POWER SUPPLY KIT OPERATES UP TO A 15 mW LASER, INCLUDES ALL COMPONENTS AND PRINTED CIRCUIT BOARD, ALL YOU HAVE TO DO IS STUFF AND SOLDER THE CIRCUIT BOARD . 4" BY 3" BY 3", PRICE $75
ITEM HENEU12 1 TO 1.5 MW HE-NE LASER 632.8 nM INCLUDES 12VDC INPUT POWER SUPPLY ALL IN A PLASTIC HOUSING 6.25 IN. BY 1.375IN BY 2.25 IN. TEMOO,RANDOM POL. ,1.7 MR DIVERGENCE. 12 MONTH WARRANTEE , PRICE $45
ITEM MELU12 1 TO 2 mW HE-NE LASER 632.8 NM , PULLS FROM MEDICAL EQUIPMENT .EACH UNIT INCLUDES HE-NE HEAD AND POWER SUPPLY[110VAC INPUT]. ALL YOU NEED TO PROVIDE IS A POWER CORD AND A FUSE TO MAKE THE UNIT OPERATIONAL. THE BEAM IS TEM00, POLORIZED WE WILL COVER EACH UNIT WITH A 12 MONTH UNLIMITED HOUR WARRANTEE, EXCELLENT FOR FOR LAB OR HOME USE. NEW THESE COST APPROX. $350 OUR PRICE $85. DIMENSIONS 9.75 BY 1.25 INCHES, P.S. 4.25 BY 3.25BY 1.25 INCHES.
ITEM RAMCNS1: RAMAN CELL OPTICS 308 nm AR/AR 4600 A 0=0 DEGREES 1000 MM FL. 2" DIA. NEW. ORIGINAL PRICE $520 OUR PRICE $175
ITEM TFPOLNS1: POLARIZERS , THIN FILM FOR 532 nm , NEW, ORIGINAL COST $590 EACH OUR PRICE $200 EACH 10 MM DIA.
ITEM CO2OCNS1: CO2 HIGH REFECTOR AND OUTPUT COUPLER 10.5 MM DIA, OC =79%R NEW. $200 A SET.
ITEM 25MNS1: DIELECTRIC BROADBAND MIRRORS 450 TO 700NM , NEW WITH PLASTIC PROTECTIVE COATINGS , 2 SIZES 25 MM SQ. AND 50 MM SQ. RECOMENDED FOR HIGHER POWER LASERS.
ITEM # BSDNS1: 50/50 DIELECTRIC COATED PLATE BEAM SPLITTER 630 TO 660 NM COMES IN A TRIANGLE SHAPE EACH SIDE APPROX. 1" PRICE $20
ITEM # 45NS1 45 DEGREE RED REFLECTOR , PASSES 488 TO 532NM , CAN BE USED TO COMBINE RED AND GREEN/BLUE LASERS TO CREATE A WHITE LIGHT LASER. 1" SQ. PRICE $15
ITEM# PCINS1 PLANO/CONVEX LENS COATED FOR YAG 1064NM , AR COATED, 10MM DIA. NEW, ORIG. COST $250 OUR PRICE $100
ITEM# INFILTER : INTERFERENCE FILTERS USED FOR PASSING A PARTICULAR SPECTRAL LINE , 11.8 MM DIA. CAREFULLY REMOVED FROM MEDICAL EQUIPMENT AND WRAPPED IN LENSE PAPER. THE FOLLOWING WAVE LENGTHS ARE AVAILABLE. 523.5, 547.4 , 572.1, 512.9, 550.6, 488, 505.7 nm price $20 each.
FOR A COMPLETE LINE OF NEW AND USED LASERS - OPTICS -ELECTRO OPTICS- LASER SHOWS ORDER A COMPLETE CATALOG AT MWKINDUSTRIES.COM
TO: ORDER GO TO OUR WEB SITE MWKINDUSTRIES.COM {SECURE ORDERING SITE}
QUESTIONS OR REMOVAL FROM MAILING LIST EMAIL: MWK-at-WORLDNET.ATT.NET
I am interested in visualising cellulose microfibrils and microtubules/microfilaments in wood-forming cells of trees in the confocal microscope. I use FITC-labelled antibodies for cytoskeleton and would like to stain the cellulose with a fluorescent dye. If I had access to a UV laser I would have no hesitation in using calcofluor/tinopal, but I don't. So, can anybody suggest a visible light-excited fluorochrome that will localise the cellulose and will permit imaging of both cellulose and cytoskeleton? (If it helps we have a Zeiss 510 with 488, 568 and 633 nm lines.)
I thank you in advance,
Yours sincerely,
Nigel Chaffey
----------------------------------------------------- Dr Nigel Chaffey, Dept Forest Genetics & Plant Physiology, Swedish University of Agricultural Sciences, S-901 83 Ume=E5, Sweden Phone: +46-90-786-6305 =46ax: +46-90-786-5901 eMail: nigel.chaffey-at-genfys.slu.se
I am interested in visualising cellulose microfibrils and microtubules/microfilaments in wood-forming cells of trees in the confocal microscope. I use FITC-labelled antibodies for cytoskeleton and would like to stain the cellulose with a fluorescent dye. If I had access to a UV laser I would have no hesitation in using calcofluor/tinopal, but I don't. So, can anybody suggest a visible light-excited fluorochrome that will localise the cellulose and will permit imaging of both cellulose and cytoskeleton? (If it helps we have a Zeiss 510 with 488, 568 and 633 nm lines.)
I thank you in advance,
Yours sincerely,
Nigel Chaffey
----------------------------------------------------- Dr Nigel Chaffey, Dept Forest Genetics & Plant Physiology, Swedish University of Agricultural Sciences, S-901 83 Ume=E5, Sweden Phone: +46-90-786-6305 =46ax: +46-90-786-5901 eMail: nigel.chaffey-at-genfys.slu.se
Below is the result of your feedback form. It was submitted by (Barbara308-at-aol.com) on Saturday, December 6, 1997 at 20:24:01 ---------------------------------------------------------------------------
Email: Barbara308-at-aol.com Name: Barbara Oakley
School: Oakland University
State: Michigan
Zip: 48317
Question: Dear Madame or Sir,
My name is Barbara Oakley--I'm a grad student at Oakland University in Rochester, Michigan. I've been given a budget of $25,000 to buy a new metallograph for our material science laboratory. Can you give me any advice? Right now I'm looking at the PME3 from Leco....
Below is the result of your feedback form. It was submitted by (bhaab-at-zinc.cchem.berkeley.edu) on Friday, October 31, 1997 at 14:12:11 ---------------------------------------------------------------------------
Email: bhaab-at-zinc.cchem.berkeley.edu Name: Brian B. Haab
School: U.C. Berkeley
State: CA
Zip: 94720
Question: Hi,
My question has to do with imaging and the thickness of the cover plate used. How important is it to use a cover plate thickness for which the objective was specifically designed? For example, if I'm using an objective which says 0.17 (presumably corrected for a 170 micron cover slip thickness), would image quality be greatly distorted when using something, say, twice as thick? Also, what is the definition of "working distance?"
Louis DeFilippi (by way of Nestor J. Zaluzec) wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello!: I was wondering if one of your associates might help me. I have an } older (20-30 years) microscope. It is a Spencer/American Optical, bifocal, } oil immersion, serial number 155715, but with no other type number. The } instruction manual was lost (prior owner) many years ago. Is there anyone } out there who might have a copy of the manual that they might share with an } ignorant biochemist? Please help. E Mail defilip1-at-flash.net. Thank you } } Louis Dear Louis,
I have some older information on AO microscopes. If you can send a more complete description (size, color, any other markings on objectives, stand, etc.), I will try to see if it matches any of our literature.
Even better, if you can send a Polaroid print to our offices.
Thanks, Barbara Foster Consortium President Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108-2838 USA PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com **************************************************** Microscopy/Microscopy Education America's first consortium of microscopy experts offering customized on-site training & applications solutions
I am trying to make a TEM specimen of Diamond, I have a Gatan PIPS at my disposal, but to use it my Diamond wafer has to be less than 100 microns thick. At the it is 1mm thick. Any suggestions on how to thin the Diamond wafer? The Diamond is actually poly-crystaline CVD
Thank you in advance Adam Dr Adam Papworth Dept Materials science & Engineering Ashton Building The University of Liverpool L69 3BX
Phone No 0151 794 5372 Fax No 0151 794 4675 E-Mail adamp-at-liv.ac.uk
Hello Michel...are you evaporating Carbon or a Pt-C mixture for your replicas? I can only comment on my experience using Pt-C. If your buffer has changed, this may affect the ability of the replica to release. High salt or EDTA causes difficulties. We routinely use 0.1 M ammonium bicarb or 1% acetic acid. You might try exposing the mica to the vapors of 1% acetic acid, in a closed container, following evaporation. This works wonders for releasing otherwise difficult replicas.
Good Luck,
Doug Keene Shriners Hospital Research ---------------------- Doug Keene DRK-at-shcc.org
} My question has to do with imaging and the thickness of the cover plate used. } How important is it to use a cover plate thickness for which the objective } was specifically designed? For example, if I'm using an objective which } says 0.17 (presumably corrected for a 170 micron cover slip thickness), } would image quality be greatly distorted when using something, say, twice } as thick?
The cover plate thickness is the distance between the top of the cover plate and the specimen so it includes the mounting medium AS WELL AS the thickness of the cover plate itself. The refractive index of the mounting medium can therefore be quite important. AND the depth of the mount.
If the mount/plate is twice as thick you will see severe distortion of the image with the periphery quite out of focus.
Cover plate thickness does not much affect oil immersion lenses as the oil optically bonds the front element of the lens to the plate. For this reason all 100x and some very good 63x and 40x objectives are used with oil immersion. But it is very important for "high dry" lenses where you are hoping for good images. The best high dry (63x, 40x) objectives have a correction collar which you can adjust to compensate for differing cover plate/mounting medium thickness. If you don't have one of these, you have to be careful to use a cover plate of the thickness the lens is corrected for
Also, what is the definition of "working distance?"
Its the distance between the front lens of the objective and the top of the cover plate.
} } } } } --------------------------------------------------------------------------- } } } } Mel Dickson Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
I received this directly from Dennis. It should interest anyone who is doing microscopy outreach, particularly if it's with SEM, so I'm taking the liberty of placing it on the listserver:
Dear Friends and Associates, Discovery Channel's documentary series, "Movie Magic", is airing a segment called "Far Out Creatures" that features special effects used in the making of science fiction films (Alien Resurrection and Starship Troopers). Part of the 1/2 hour segment will include a short interview with me and some of my "MicroAliens". I thought this might be of interest to you.
The segment will run: December 11, 1997 9:30 - 10:00 PM - West Coast Time December 12, 1997 1:30 - 2:00 AM - West Coast Time December 13, 1997 2:00 - 2:30 PM - West Coast Time
Please check your local listing for the time of Movie Magic on the Discovery Channel for these days.
Best Regards, Dennis Kunkel
*********************************************** * Dennis Kunkel Ph.D. * * Pacific Biomedical Research Center * * University of Hawaii * * * * email - kunkel-at-pbrc.hawaii.edu * * www - http://www.pbrc.hawaii.edu/~kunkel/ * ***********************************************
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
bhaab-at-zinc.cchem.berkeley.edu wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Below is the result of your feedback form. It was submitted by } (bhaab-at-zinc.cchem.berkeley.edu) on Friday, October 31, 1997 at 14:12:11 } --------------------------------------------------------------------------- } } Email: bhaab-at-zinc.cchem.berkeley.edu } Name: Brian B. Haab } } School: U.C. Berkeley } } State: CA } } Zip: 94720 } } Question: Hi, } } My question has to do with imaging and the thickness of the cover plate used. } How important is it to use a cover plate thickness for which the objective } was specifically designed? For example, if I'm using an objective which } says 0.17 (presumably corrected for a 170 micron cover slip thickness), } would image quality be greatly distorted when using something, say, twice } as thick? Also, what is the definition of "working distance?" } } Thank you very much, } } Brian Haab } } --------------------------------------------------------------------------- Dear Brian, Coverslips are considered part of the optical design of a microscope. If the objective carries the marking "0.17", it expects to see a coverslip of approximately that thickness on your sample. While it may tolerate a slight deviation (0.15-0.18), the image may be seriously degraded outside those limits. By the way, as you might have noticed, there is very little information in the catalogs relating this thicknessness to the ordring specifications. For 0.17mm, order a #1 1/2 coverslip.
Regarding working distance: it is open distance or space between the front lens of the objective and the top of the preparation. Condensers also have working distances": from the top element of the condenser to the bottom of the slide/preparation.
We cover all of this and much more in our book, "Optimizing Light Microscopy for Biological and Clinical Laboratories". If you would like to order one, email me and we will forward an electonic order form.
Hope this helps.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108-2838 USA PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com **************************************************** Microscopy/Microscopy Education America's first consortium of microscopy experts offering customized on-site training & applications solutions
Barbara308-at-aol.com wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Below is the result of your feedback form. It was submitted by } (Barbara308-at-aol.com) on Saturday, December 6, 1997 at 20:24:01 } --------------------------------------------------------------------------- } } Email: Barbara308-at-aol.com } Name: Barbara Oakley } } School: Oakland University } } State: Michigan } } Zip: 48317 } } Question: Dear Madame or Sir, } } My name is Barbara Oakley--I'm a grad student at Oakland University } in Rochester, Michigan. I've been given a budget of $25,000 to buy a new } metallograph for our material science laboratory. Can you give me any } advice? Right now I'm looking at the PME3 from Leco.... } } I'd appreciate any help you could provide. } } Barb Oakley } } --------------------------------------------------------------------------- Dear Barb,
Most of the microscope companies carry superb metallographs. What do you need in the way of specific functionality (i. e., camera ports, magnification, upright vs. inverted, ability to project reticles for visual measurement, etc.).
In your budget range, you probably would do well to consider the Olympus (sold through LECO) or a Unitron system. Another alternative is a good quality used Reichert MEF3 or MEF4. If you need contacts, phone numbers, etc., please email me.
Send me a copy of your specifications and applications and I will see who else might have what you are looking for.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108-2838 USA PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com **************************************************** Microscopy/Microscopy Education America's first consortium of microscopy experts offering customized on-site training & applications solutions
Valdre' Andrea wrote: } } } I' tryng to collect all info possible about "Image Plates" devices. } } I know that FUJI had in the past a sytem called FDL 5000. } (I have a 1 page leaflet). Fuji, in Italy does know nothing about this produc. } Can anyone help me with more info, approx. price, where to find it, etc.
Try http://home.fujifilm.com/info/products/science/ip/index.html I hope that helps.
Philip -- Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 86 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www_scem.csb.ki.se/pages/philip.html
Does anyone know of any commercially available ABs that react with cytoskeletal elements in cereal plants? We wish to do immunofluorescence but have failed using commercially available alpha- and beta-tubulin monoclonal ABs (N356 and N357 ABs from Amersham). We have done Western blots and these do not appear to recognize the respective tubulins in rice. Any advice? Thanks.
Tim Bourett DuPont Experimental Station Wilmington, DE USA
I do not have any experience with the Leco line of microscopes but I have used others. We currently have a Nikon Epiphot that I think performs better than most of the other metallographs that I have used. Make sure you get demos and have plenty of samples to try test on them before you buy. You should alos get a box of film and record an image from each one. Good luck.
I don't know about wood but in human tissue we use a .01% Evans Blue as a total protien counterstain in conjunction with FITC tagged primary. The evans blue excites and emmits like rodamine or texas red.
Bob
On Sun, 7 Dec 1997, Nigel Chaffey wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } =20 } Dear Fellow Microscopists, } =20 } I am interested in visualising cellulose microfibrils and } microtubules/microfilaments in wood-forming cells of trees in the confoca= l } microscope. I use FITC-labelled antibodies for cytoskeleton and would li= ke } to stain the cellulose with a fluorescent dye. If I had access to a UV } laser I would have no hesitation in using calcofluor/tinopal, but I don't= =2E } So, can anybody suggest a visible light-excited fluorochrome that will } localise the cellulose and will permit imaging of both cellulose and } cytoskeleton? (If it helps we have a Zeiss 510 with 488, 568 and 633 nm } lines.) } =20 } I thank you in advance, } =20 } Yours sincerely, } =20 } Nigel Chaffey } =20 } ----------------------------------------------------- } Dr Nigel Chaffey, } Dept Forest Genetics & Plant Physiology, } Swedish University of Agricultural Sciences, } S-901 83 Ume=E5, } Sweden } Phone: +46-90-786-6305 } Fax: +46-90-786-5901 } eMail: nigel.chaffey-at-genfys.slu.se } =20 } Looking for another job/position/post... } =20 } =20 } =20
} PDC-2000 is a fine hand-held digital camera but does not have a way to } adapt for the microscope. Amont other things there are internal } optics that don't cooperate well with the microscope optics. It also } could do some macro work with close-up lenses, but that is not } optically the best imaging system.
OK. So, Polaroid removes the optics.
} DMC, while using the same Polaroid designed sensor chip as PDC-2000, } otherwise is designed and built specifically for microscopy. Its } C-mount thread (no optics) allows easy mounting on almost any } microscope using standard C-mount adapters, and also can accept many } "C-mount" threaded macro lenses for use on the copystand.
Adding a c-mount and a tripod socket is worth about $50. I fail to see how the price of the DMC is justified. This seems to be another example of how specialised users pay higher prices for less product. I know that 'economies of scale' do not apply to the technical market.
Can anyone supply me with a source of human skeletal muscle suitably prepared for IEM? Or.. would anyone be able to loan blocks of LR White embedded human skeletal muscle that I could cut sections from and return?
Brett M. Connolly, PhD Merck Research Laboratoriess email: brett_connolly-at-merck.com
I have a customer who is looking for a technique to examine a few small ( {1 mm) bubbles in quartz via Raman or other IR technique. The objective is to thin the quartz to within 1 micrometer of the bubble. The sample size is a few centimeters but can be reduced if necessary.
Any help will be appreciated.
Thanks,
Harold J. Crossman OSRAM SYLVANIA INC. Lighting Research Center 71 Cherry Hill Dr. Beverly, MA 01915 Phone: (508) 750-1717 E-mail: crossman-at-osi.sylvania.com
Our web sites: www.sylvania.com www.siemens.com --
} Please could somebody help me. } } I am trying to make a TEM specimen of Diamond, } I have a Gatan PIPS at my disposal, but to use it my Diamond wafer } has to be less than 100 microns thick. At the it is 1mm thick. } Any suggestions on how to thin the Diamond wafer? } The Diamond is actually poly-crystaline CVD } } Thank you in advance } Adam
Hi Adam, Well, I have looked at some single crystal diamond, and the results were great, this is how the sample was prepared;
The sample was sliced with a diamond saw as thinly as we could get, and we tried to dimple it but that was really pointless, we only got 30um reduction in thickness in seven days!
So we PIP's it from quite a thick sample, total time taken was 3080 minutes! Beam at 5KV rot speed 3.5 rpm Ion currents 25uA Gun angle 5'.
Your sample may not be quite as hard, so you many have less time in the PIPS. Also, I found that trying to punch a 3mm disk with a ultrasonic disk cutter is also pointless, much better to mount on a disk then grind the edges round. Be prepared to change the mounting ring several times as the beam will destroy this quicker relative to the diamond.
I hope this helps, btw. Tony Garratt-Reed says hello and sends his regards!
Cheers
David
Dr. David C. Bell Room 13-1018 E-Mail: dcb-at-MIT.EDU Center for Mat. Sci. and Eng. PH: (617) 253-3317 Massachusetts Institute of Technology FAX: (617) 258-6478 77 Massachusetts Ave, Cambridge, MA 02139-4307
Have any of you placed 160 mm objectives to a Zeiss Axioplan or Axiophot with good results? Carl Zeiss does sell an adapter that threads into many 160 mm objectives, but I would perfer to put a negative lens on a slider below the tube lens. I estimate that a -360 mm lens placed about 40 mm below the tube lens would do the trick. I would also welcome comments on the Axioplan II and the merits of the new optics.
Rob ---------------------------------------------------------------------------- ---------------------- Robert W. Williams Center for Neuroscience, Department of Anatomy and Neurobiology 875 Monroe Avenue, Memphis TN 38163 USA Tel: 901/448-7018 or -7050 FAX: -7193 http://mickey.utmem.edu/neuron.html rwilliam-at-nb.utmem.edu
Can anyone supply me with a source of human skeletal muscle suitably prepared for IEM? Or.. would anyone be able to loan blocks of LR White embedded human skeletal muscle that I could cut sections from and return?
Brett M. Connolly, PhD Merck Research Laboratoriess email: brett_connolly-at-merck.com
You might try ultramicrotomy to prepare cross-sections of diamond for TEM analysis. I've had a great deal of experience and luck cross-sectioning hard materials using the technique. You can review an ultramicrotomy procedure for cross-sectioning diamond and cBN in "Ultramicrotomy of Diamond Films for TEM Cross-section Analysis," P. Swab, Microscopy Research and Technique, Wiley-Liss Inc., vol. 31, pp. 308-310 (1995) and other references found in the paper. The paper describes the formation of "micro chips" that are preferentially oriented, embedded in epoxy, and then cross-sectioned with a diamond knife. These cross-sections may show mechanical artifacts, but are uniformly thick and free of beam damage and chemical artifacts.
The concoidal micro chips generated in this procedure are very thin at the edges and may be sufficiently thin for direct TEM observation. If not, the microchips may be secured to a grid and thinned using conventional ion beam techniques. Ion-thinned cross-sections show a minimum of mechanical artifacts, but are not uniformly thick and may show beam damage and associated chemical artifacts.
For assistance with ultramicrotomy in the UK, contact John Forsdyke at Oxford Brookes University (jforsdyk-at-bms.brookes.ac.uk).
Regards,
Phil Swab Advanced Coatings Division Advanced Refractory Technologies Buffalo, NY, USA Phone: 716-875-4091 E-mail: pswab-at-art-inc.com
} ---------- } From: Adam Papworth[SMTP:A.J.Papworth-at-LIVERPOOL.AC.UK] } Sent: Sunday, December 07, 1997 3:37 PM } To: microscopy-at-Sparc5.Microscopy.Com } Subject: TEM of Diamond } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } ---------------------------------------------------------------------- } -. } } Please could somebody help me. } } I am trying to make a TEM specimen of Diamond, } I have a Gatan PIPS at my disposal, but to use it my Diamond wafer } has to be less than 100 microns thick. At the it is 1mm thick. } Any suggestions on how to thin the Diamond wafer? } The Diamond is actually polycrystalline CVD } } Thank you in advance } Adam } Dr Adam Papworth } Dept Materials science & Engineering } Ashton Building } The University of Liverpool } L69 3BX } } Phone No 0151 794 5372 } Fax No 0151 794 4675 } E-Mail adamp-at-liv.ac.uk } }
It is not appropriate to share human tissue. Each project should be justified according to the local and national regulations. Most importantly the people (or their survivors) who provided the tissue should be informed and allowed to decline participation in the project. I hope that this request does not represent the state of ethics at all commercial organizations! Larry D. Ackerman (415) 476-8751 Howard Hughes Medical Institute FAX (415) 476-5774 UCSF, Box 0724, Rm U426 533 Parnassus Ave. mishot-at-itsa.ucsf.edu San Francisco, CA 94143
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com
I am trying to make a TEM specimen of Diamond, I have a Gatan PIPS at my disposal, but to use it my Diamond wafer has to be less than 100 microns thick. At the it is 1mm thick. Any suggestions on how to thin the Diamond wafer? The Diamond is actually poly-crystaline CVD
Thank you in advance Adam Dr Adam Papworth Dept Materials science & Engineering Ashton Building The University of Liverpool L69 3BX
Phone No 0151 794 5372 Fax No 0151 794 4675 E-Mail adamp-at-liv.ac.uk
Adam:
You might try ultramicrotomy to prepare cross-sections of diamond for TEM analysis. I've had a great deal of experience and luck cross-sectioning hard materials using the technique. You can review an ultramicrotomy procedure for cross-sectioning diamond and cBN in "Ultramicrotomy of Diamond Films for TEM Cross-section Analysis," P. Swab, Microscopy Research and Technique, Wiley-Liss Inc., vol. 31, pp. 308-310 (1995) and other references found in the paper. The paper describes the formation of "micro chips" that are preferentially oriented, embedded in epoxy, and then cross-sectioned with a diamond knife. These cross-sections may show mechanical artifacts, but are uniformly thick and free of beam damage and chemical artifacts.
The concoidal micro chips generated in this procedure are very thin at the edges and may be sufficiently thin for direct TEM observation. If not, the microchips may be secured to a grid and thinned using conventional ion beam techniques. Ion-thinned cross-sections show a minimum of mechanical artifacts, but are not uniformly thick and may show beam damage and associated chemical artifacts.
For assistance with ultramicrotomy in the UK, contact John Forsdyke at Oxford Brookes University (jforsdyk-at-bms.brookes.ac.uk).
Regards,
Phil Swab Advanced Coatings Division Advanced Refractory Technologies Buffalo, NY, USA Phone: 716-875-4091 E-mail: pswab-at-art-inc.com
Hi everyone, Would anyone have the address / phone number for HITEK Cryogenic Engineering ?The last address I have is: 3190 Park Road Bay Vista Business Park Benicia, CA 94510
TIA Rosemary
#################################################### Rosemary Walsh Electron Microscope Facility for the Life Sciences The Biotechnology Institute for Research and Education 1 South Frear Lab University Park, PA 16802 814-865-0212 email:rw9-at-psu.edu ####################################################
I can only suggest that, if you are sincerely interested in a high quality digital camera for your microscope, you evaluate the DMC first-hand and compare it with similar products, most of which cost thousands of dollars more. Compare, also, some of the lower-cost solutions and adapters available and determine what image quality level you truly require. Then buy whatever meets your needs at the lowest cost to you.
There is significant investment involved when developing a product specifically for a specialized market, such as optical microscopy. Companies, Polaroid included, need to recover their investment costs so that they can continue to serve their customers.
If you'd like to discuss further, or if you'd like to arrange a product demonstration locally, please get back to me directly.
I hope this helps you in your concerns and that we are able to be of service to you. Thanks for your interest!
} PDC-2000 is a fine hand-held digital camera but does not have a way to } adapt for the microscope. Amont other things there are internal } optics that don't cooperate well with the microscope optics. It also } could do some macro work with close-up lenses, but that is not } optically the best imaging system.
OK. So, Polaroid removes the optics.
} DMC, while using the same Polaroid designed sensor chip as PDC-2000, } otherwise is designed and built specifically for microscopy. Its } C-mount thread (no optics) allows easy mounting on almost any } microscope using standard C-mount adapters, and also can accept many } "C-mount" threaded macro lenses for use on the copystand.
Adding a c-mount and a tripod socket is worth about $50. I fail to see how the price of the DMC is justified. This seems to be another example of how specialised users pay higher prices for less product. I know that 'economies of scale' do not apply to the technical market.
Can anyone tell me about how often a diamond knife needs to be sharpened? I realize it would depend on the amount of wear it gets. Our EM dept. is only open three days a week, with cutting being done about 2-4hrs a week. Our knife was sharpened six months ago and seems to be getting dull again. Is this to be expected?
I know that there are MAC and PC versions of NIH Image but has anyone used NIH Image on a Silicon graphics system? If so, would you please tell me what is needed to set it up.
Salzburg, 12/9/97 Dear Brian,=20 unfortunately I do not know wether you got my reply to your posting on = that=20 subject correctly by 11/02/97 as you can see as follows: =20
Salzburg, 11/02/97
---------- Von: azriel gorski[SMTP:azrielg-at-cc.huji.ac.il] Gesendet: Sonntag, 02. November 1997 09:24 An: David S. Wexler, Ph.D. Cc: Microscopy-at-sparc5.microscopy.com Betreff: Re: cover slip thickness
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
On Fri, 31 Oct 1997, David S. Wexler, Ph.D. wrote:
} = ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of = America=20 } To Subscribe/Unsubscribe -- Send Email to = ListServer-at-MSA.Microscopy.Com } = -----------------------------------------------------------------------. } =20 } Hi, } =20 } I have a question about the importance of cover slip thickness. = Namely, } how important is it to use a cover slip thickness for which the } objective } is designed? For example, if I'm using a 40X, NA 1.3 oil immersion } objective which has the number 0.17 on it (corrected for a 170 micron } cover slip thickness), what would be the effect on image quality if I } used a cover slip with, say, a 300 micron thickness?
} Brian Haab 11/02/97 wrote: } U.C. Berkeley } bhaab-at-zinc.cchem.berkeley.edu } =20
} Good luck,
} Shalom from Jerusalem, } Azriel=20
} ******************************************************** } Azriel Gorski, Head azrielg-at-cc.huji.ac.il } Optical Microscopy Laboratory } Division of Identification and Forensic Science } Israel National Police } Jerusalem } ISRAEL
} It is important. You are using a precision optical instrument and } the cover slip is an indespensible part of the optical correction. = Since=20 } you are using an oil immersion objective at 40 X (with oil I hope) it } appears you want as much defined and clear detail as posible. Using any } lense above about 40 X you can see the difference between #1 cover = slips } (0.13 to 0.17mm thick), #1 1/2 coverslips (0.16 to 0.19mm thick), and = #2=20 } cover slips (0.17 to 0.25mm thick). Now having said that, there is an } assumption implicit in that. That is that the sample adhears directely = to } the underside of the coverslip. So don't "flood" with mounting medium.
W. MUSS writes: The cover slip for "dry" objectives with a numeric aperture N.A. below /up to 0.3/0.4 is said to be no problem in terms of optic geometry as well as image quality. You can use "dry"objectives with an N.A. {=20 0.40 with or without a coverslip. For "dry" objectives with an N.A. of=20 =3D/} 0.4 the cover slip is part of the "optical correction" system of = the=20 optical lenses. If you don=B4t use such cover slips, due to spherical=20 aberration you will get diminished contrast quality. The higher the N.A of the objective will be, the smaller will be the tolerances: for example:
Numerical Aperture (NA): effective thickness of cover slip: 0.40 0.17 +/- 0.09 mm 0.60 0.17 +/- 0.013 mm 0.75 0.17 +/- 0.004 mm
These theoretical values not necessarily don=B4t apply for the reality: They are valid assuming tissue sections adhere closest to the lower=20 side of the coverslip. In practice between upper side of the tissue=20 section and the lower side of the cover slip there is a small layer of=20 mounting medium, the actual thickness you won=B4t know. So, in=20 practice you should/can use a minimum of mounting medium, and cover slips measuring 0.15-0.16 mm. If using N.A.=B4s } 0.75 you = won=B4t be able to reach the goal "0.17" precisely. Therefore you can choose dry objectives with a "correction collar" which allows you to adjust = the optical correction between 0.12 and 0.22 mm.
With "immersion objectives" it seems to be more critical: For obtaining optimal resolution and contrast =3D optimal image quality you should be aware of the quality of the immersion oil, of the=20 coverslip, of the mounting medium as well as of the temperature. Immersion oil, and the coverslip then will be a PART of THE OBJECTIVE itself. An immersion oil should have at 23 degr. C a=20 refractive index "n lower case e" of 1.518 (the dispersion, "Abbe=B4s numeral" not considered here). Coverslips for oilimmersion-objectives mostly are obligatory.=20 Deviation/variations of thickness of the coverslips here wouldn=B4t be that critical than with the "dry objectives" (see above): compensating for that will work the drop of immersion oil between the cover slip and the objective lens. An other point worth to be considered with respect to that would be variation(s) in the refractive power. So, as a "theoretical" consequence one should use cover slips with the same refractive index of the mounting medium/immersion medium. Normally, you=20 won=B4t get the information about the refractive index of cover slips, = or do you??
But anyway, you will get optimal results only (especially if using an objective with NA of 1.40) when using additionally an immersion-oil- condensor lens (because oil immersion with its high NA needs a=20 corresponding illumination aperture: if you want to benefit from the = full=20 performance of your optical system the illumination aperture =3D condensor lens has to be brought up to the objective aperture =3D objective lens. The immersion-condensor should have a similar big aperture than the objective.)
} As an asside, I know one microscopist who measures his coverslips with = a } micrometer and only uses the ones which are actually 0.17mm.
W.MUSS writes: You can buy also coverslips which are proved to be 0.17 +/- 0.01 mm as well as other qualities which are the more expensive the less they exhibit variation from 0.17 mm. But, as Brian proposes, I too know LM-freaks measuring a package of "normal" coverslips one by one with a micrometer. This may be the cheapest way in being sure about 0.17 mm=20 thickness.
------------------------------------------------------=20 } Also, what is the definition of "working distance?" I understand it = to } be the distance between the top of the cover slip and the lens of the } objective, but I want confirmation of this definition.
} You are basically correct.... to be a "sticler" it is the nearest part = of } the lense, not any optical center of it.
W.MUSS writes: The higher the magnification number of an objective, the smaller the=20 "working distance" will be. Besides problems in positioning a high power objective carefully over a section/cover slip and thereby without damaging it (which wouldn=B4t be possible with modern objectives with=20 a "sliding/retractive" mechanism of the objective itself) you certainly =
will have or can get problems with an increased thickness of your cover slip (see above): say, working distance of your } N.A. 1.40, 40x=20 Oil Immersion Objective { will be 0.2 mm (see info-sheet of your=20 microscope's or objective's instruction manual, you *can* use a=20 coverslip 0.3 mm thickness: but then you will not be able to "optically reach" the tissue section for correct imaging. At least you will decrease resolution and contrast capacity. =09 Another suggestion: why not try a 40x or 50x objective with an N.A. of 1.00? This would be a compromise with respect to working distance as well as not necessarily the "must" of using immersion illumination/ immersion of the condensor front lens to achieve resolution/contrast=20 conditions as provided and possible by the optical system you use.
Also: if you view semithin sections with your scope, you won=B4t use = cover=20 slips at all: try viewing directly with a drop of immersion oil on your =
sections and the oil immersion objective. You will get marvellous image quality (provided you have adjusted your optical system according to KOEHLER=B4s instructions). Removal of the immersion oil is just immersion of the object slide(s) into a coplin jar containing xylene. After soaking a while you can wipe off (or blow off by use of compressed air) very easily remnants of immersion oil.
Hope this explanation helps you with your work best regards "on a lazy sunday 2nd of november from SALZBURG- Mozart=B4s birthplace"
Wolfgang MUSS the same on Tue, 9th Dec. 1997 Department of Pathology, LKA EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
---------- Von: bhaab-at-zinc.cchem.berkeley.edu[SMTP:bhaab-at-zinc.cchem.berkeley.edu] Gesendet: Sonntag, 07. Dezember 1997 18:29 An: microscopy-at-sparc5.microscopy.com Betreff: LM: imaging and the thickness of the cover plate
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
Below is the result of your feedback form. It was submitted by (bhaab-at-zinc.cchem.berkeley.edu) on Friday, October 31, 1997 at 14:12:11 -------------------------------------------------------------------------= --
Email: bhaab-at-zinc.cchem.berkeley.edu Name: Brian B. Haab
School: U.C. Berkeley
State: CA
Zip: 94720
Question: Hi,
My question has to do with imaging and the thickness of the cover plate = used. How important is it to use a cover plate thickness for which the = objective was specifically designed? For example, if I'm using an objective which says 0.17 (presumably corrected for a 170 micron cover slip thickness), would image quality be greatly distorted when using something, say, = twice as thick? Also, what is the definition of "working distance?"
The DMC and any other product of its kind are worth what people will pay for it. It is the nature of the free market economy. You must consider=
the product development costs, which are substantial. Also, how many wil= l they sell? The market size is not huge.
The Polaroid DMC brought was introduced at about $6000. This is FAR less=
than any other quality digital microscope camera on the market. The only=
less expensive options come from something like the Pixera, which is not realistic for anyone who wants to get true high quality images without massive pixel enlargement. It seems odd to blast Polaroid for introducin= g a camera that was $5000 less than their nearest competitor when introduce= d. The good news is that prices always go down, and quality goes up.
} The DMC and any other product of its kind are worth what people will pay } for it.
Undeniable.
} The Polaroid DMC brought was introduced at about $6000. This is FAR less } than any other quality digital microscope camera on the market. The only } less expensive options come from something like the Pixera, which is not } realistic for anyone who wants to get true high quality images without } massive pixel enlargement. It seems odd to blast Polaroid for introducing } a camera that was $5000 less than their nearest competitor when introduced.
That's not the thrust of my argument. Why is a camera with fewer features/components so much more expensive than the more complex one? It seems less of a market-driven issue than a marketing one, similar to the observation that I can buy DuMont forceps from a jeweler's supplier for half the cost as from a medical supplier.
} The good news is that prices always go down, and quality goes up. } }
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
It also depends on what is being cut as well as who is doing the cutting. If it is plant material, 6 months is a LONG time. Animal tissue is = softer but again some animal tissues have hard components which are hard = on the knife edge. The experience of the cutter obviously also comes into play. The less = experience, usually the more resharpenings may be necessary. Are they = also taking thicks on the same knife? If so, you might want to get a = histo knife for thicks and perhaps your thin diamond would last longer. Good Luck Judy Murphy San Joaquin Delta College Microscopy Technology Center Stockton, CA __________________________________________________________________________= _____
Can anyone tell me about how often a diamond knife needs to be sharpened? = I realize it would depend on the amount of wear it gets. Our EM dept. is = only open three days a week, with cutting being done about 2-4hrs a week. Our knife was sharpened six months ago and seems to be getting dull again. Is this to be expected?
------------------ RFC822 Header Follows ------------------ Received: by sjdccd.cc.ca.us with ADMIN;9 Dec 1997 08:58:05 -0800 Received: from Sparc5.Microscopy.Com (206.69.208.10) by = ms.sjdccd.cc.ca.us with SMTP (Eudora Internet Mail Server 1.2); Tue, 9 Dec 1997 08:57:56 = -0800 Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com = (8.6.11/8.6.11) id HAA12949 for dist-Microscopy; Tue, 9 Dec 1997 07:56:36 = -0600 Received: from [206.69.208.21] (mac21.zaluzec.com [206.69.208.21]) by = Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id HAA12946 for = {microscopy-at-msa.microscopy.com} ; Tue, 9 Dec 1997 07:56:32 -0600 X-Sender: zaluzec-at-microscopy.com Message-Id: {v03007803b0b301d55bd4-at-[206.69.208.21]} Mime-Version: 1.0 Content-Type: text/plain; charset=3D"us-ascii"
} Does anyone know of a quick test that would confirm pen ink (Flair, Bic, } etc.) on paper? I have plenty of sample that consists of large, black, } blue-fringed blotches on a flooring felt backing. There are no visible } pigment particles up to about 400X, and the stain is moderately soluble in } an ethanol/methanol blend. } } Thanks in advance.
David, you might find it helpful to submit this question to the forensic science list. To submit, send the message to forens-l-at-ACC.FAU.EDU. To subscribe, send a message to MailServ-at-Acc.Fau.Edu with the following in the body of the message: Subscribe Forens-L Your Real Name.
This is just the kind of thing the forensic people do all the time.
-- ********************************************************** Stephen A. Shaffer sshaffer-at-microdataware.com MicroDataware http:www.microdataware.com (Under reconstruction and temporarily out of service) Personal stuff: steve_shaffer-at-compuserve.com http://ourworld.compuserve.com/homepages/steve_shaffer/ **********************************************************
does anyone have a good source for transfer roles and paper for a tektronix Phaser 440 dye sub printer?
thanks
steve
--------------------------------------------------------------------- Dr. Steven Barlow, Associate Director EM Facility/Biology Department 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/
Dear Diane, have found your posting on an issue which only can be answered very subjectively. For further elucidation one should know not only weekly/daily hours of usage but, more interestingly:
- what company product ( e.g. DIATOME, DEKKER, MICROSTAR.........) - which type of Diamond knife (e.g. HISTO-Type for semithin sections, ULTRA-Type for ultrathin sections.....) - length of cutting edge - proper working (cutting) as usually done (starting cutting from left to right edge) - max. width of specimens you are cutting - which kinds of tissues (with incrustations, artificial inclusions,...any to be expected?) - which type of, of which hardness resin(s) you use - which person will perform cutting (I understand that this will be an experienced person, familiar with cutting with diamond knife(-ives) - cutting angle, knive free angle, cutting speed: is this done correctly as implicated by the manufacturer's (guaranteed) data sheets on cutting properties - cleaning methods used (sometimes remnants of resin and other cutting debris imparts cutting quality: have a careful look on your cutting edge at least at mag. x 45, seen with back light)
If you can provide some more details according to this items, maybe there is a more objective answer. Best regards Wolfgang
Dr. Wolfgang MUSS Department of Pathology, LKA EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
DISCLAIMER: No commercial interest in products/product lines, company/-ies, if such names are mentioned or such are refered to.
---------- Von: Diane M. Smith[SMTP:smithde-at-valunet.com] Gesendet: Dienstag, 09. Dezember 1997 15:10 An: microscopy-at-sparc5.microscopy.com Betreff: Q: Diamond Knife-resharpening
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Can anyone tell me about how often a diamond knife needs to be sharpened? I realize it would depend on the amount of wear it gets. Our EM dept. is only open three days a week, with cutting being done about 2-4hrs a week. Our knife was sharpened six months ago and seems to be getting dull again. Is this to be expected?
In my initial response to this thread, I extolled the virtues of the = Kodak DC120 camera for photomicroscopy based on trials at our lab. = Maybe I should have given the standard disclaimer that I have no = affiliation with Kodak ... I definitely should have listed the other = cameras I considered.=20 Unless anyone would like to include another product, the three similar = choices mentioned in this thread are the Pixera, the Polaroid DMC, and = the soon to be released Kodak MDS120 system (based on the DC120). = Obviously, this list excludes more capable, more expensive cameras. The = question of interest is how capable are these cameras and are they = capable enough... beyond being just teaching and demonstration systems. Seth Grotelueschen makes a good point about not blasting Polaroid for = being the first to release a relatively low cost system that is = comparable to much more expensive systems. Hopefully, we are at a point = where prices for these systems continue to fall precipitously toward = affordability... and hopefully before Kalman Rubinson gets a chance to = retire : )
A quick comparison of the 3 systems follows with questions or comments = related to capability:
1.) PRICE Pixera $1200 Polaroid DMC $6000 Kodak MDS120 $2100 2.)CCD SIZE Pixera 1/3" Polaroid DMC 12.15mm(~1/2") Kodak MDS120 1/2" Question: Is the physical size of the CCD array important or is = resolution more important to image quality? 3.) CCD RESOLUTION Pixera - ~750k=09 Polaroid DMC ??? claimed to be "megapixel" Kodak MDS120 836K 4.) IMAGE RESOLUTION Pixera 1 meg =20 Polaroid DMC 1.9 meg (1600x1200) Kodak MDS120 1.2 meg (1280x960) All these systems use integration and averaging to increase final image = resolution. Question: Does this type of claim based on integrated = resolution overstate the capabilities of the camera? Are these claims = analogous to "false magnification"? Is resolution in the = photomicroscopy as important as other issues such as low light = sensitivity and data integrity? 5.) DATA INTEGRITY It is my understanding that all of these systems are capable of transferring images uncompressed to PC or Mac or loss-less compression = such as Tiff formats. As Dr. John Russ pointed out, many comsumer grade = digitals store images using lossy compression algorithms.=20 6.) IMAGE TRANSFER TO PC- all systems are TWAIN compliant Pixera - limited to speed of serial port =20 Polaroid DMC - faster SCSI transfer rate Kodak MDS120 - limited to speed of serial port=20 7.) FLEXIBILITY OF CAMERA Both the Polaroid and the Pixera are tethered to the PC and are = generally dedicated to the microscope. They can be used with a c-mount = video lens (sold separately) for other applications but must remain = tethered to a computer.=20 The Kodak camera can be used as a stand alone digital camera away from = the computer. No addition lenses are required. 8.) Strengths - all systems promise to be user friendly compared to a = video camera/ frame grabber solution=20 Pixera - Most affordable =20 Polaroid DMC - Highest resolution and speed of transfer Kodak MDS120 - Most Versatile, affordable, and good resolution
All the information above was taken from the respective spec. sheets for = each system.=20
For my application, I am trying to capture images allowing me to = optimize my probing protocols which will later be used on the confocal. = So rather than looking for "analyzable" images I am more concerned with = optimizing for analyzable images. That doesn't mean that these systems = are not capable of this. Hopefully, someone will take the time to = explain how/why these systems are/are not capable of generating = "analyzable" images
Kevin Brent Smith=20 Graduate Student University of Louisville Biology Dept. Life Science Bldg. Rm.#12 Louisville, KY 40292 Phone: (502) 852-6773 Fax: (502) 852-0725 =20
I remember there have been reports since many years ago regarding to the alternative chemical treatment in place of critical point drying using a critical point dryer. Can any of you tell the name(s) of the chemicals and the companies which sell them? Or your experience using such chemicals? Sorry for not being able to look up a SEM book for myself.
Thanks!
Yuhui Xu Core EM Facility Dana Farber Cancer Institute
We have just acquired an old Gatan Ion Mill and the Electronic Gas Flow Control (EGC) valve is not working. I am just wondering if anyone happen to have a spare EGC valve or a manual needle valve assembling to give away or for sale.
Thanks a lot.
Eric Yu Wang North Campus Electron Microbeam Analysis Laboratory 413 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (313) 936-3353 Fax: (313) 763-5567 eywang-at-engin.umich.edu http://emalwww.engin.umich.edu/people/eywang/eywang.html
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Subject: Time:3:40 PM OFFICE MEMO reply} test for ink Date:12/9/97
Reply to: RE} Test for Ink? ************************* You wrote, "Does anyone know of a quick test that would confirm pen ink (Flair, Bic, etc.) on paper? I have plenty of sample that consists of large, black, blue-fringed blotches on a flooring felt backing. There are no visible pigment particles up to about 400X, and the stain is moderately soluble in an ethanol/methanol blend. Thanks in advance. Dave Stadden" ************************* Dave, One technique might be to use X-ray fluorescence (XRF), exciting the specimens with secondary x rays. This could probably be done in a couple of hours, and doesn't require a lot of specimen prep work. XRF won't do the chemistry, but it could be very good for matching elemental content of inks (at least for those elements in the ink heavier than sodium) and their relative quantities, even in the presence of organic components. Using XRF, first look at a gross ink sample to determine elemental content and relative concentrations to optimize the type of x-ray excitation. Then collect an x-ray spectrum from the ink-on-paper specimen. Collect another spectrum (same geometry, same excitation, same acquire live time, etc) of paper only. Strip out (subtract) the paper spectrum from the ink-on-paper to get an ink signature from paper. Save. Repeat this process, subtracting the flooring felt backing "background") for the ink spectrum from the ink on felt . Compare the two spectra. If the Flair, Bic, ??, is the source of the stain, you should see a pretty good match. It won't be exact since the samples were taken from two quite different substrates. You might be able to minimize the mismatch if you prepared a "standard", using known inks on otherwise identical, unstained, felt backing. Is this an on-going analysis problem or a one-time test? Do you have access to an XRF system? Dennis DGCollins-at-lbl.gov
I'm David Langlais, Vice President of Marketing for Pixera Corporation. Since Pixera was introduced into this discussion and into discussions on some USENET news groups lately, I feel that a response to the Mr. Grotelueschen's remark is appropriate.
I'm posting from the email account of one of my employees. My email is ddl-at-pixera.com and I'll be subscribing to the listserver today so anyone can reach me with comments......
And so...
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
On Tuesday, 9 Dec 1997, Seth J. Grotelueschen wrote:
} The DMC and any other product of its kind are worth what people will pay } for it. It is the nature of the free market economy. You must consider } the product development costs, which are substantial. Also, how many will } they sell? The market size is not huge.
} The Polaroid DMC brought was introduced at about $6000. This is FAR less } than any other quality digital microscope camera on the market. The only } less expensive options come from something like the Pixera, which is not } realistic for anyone who wants to get true high quality images without } massive pixel enlargement.
Mr. Gruteleuschen and others are certainly entitled to their opinions regarding the quality of Pixera's cameras in microscopy. I just wish that he and others would take the time to be technically accurate. He and they are not. There is a common misconception and disbelief that Pixera can really achieve 1260x960 pixels in our catpured images without some nefarious trick or (gasp) 'massive pixel enlargement".
Mr. Gruteleuschen evaluated our Pixera Professional some months ago and decided not to purchase. If he chose not to purchase due to the technical reason he stated in this message, then he made his decision for the wrong re asons.... Because he either doesn't understand how our camera system works or he doesn't believe it. If the pictures he captured didn't satisfy his requirements, I would have no problem with him saying so. But his remark is technically erroneous and misleading.
Pixera's high resolution cameras - Professional and Personal, can capture 24-bit RGB color images in resolutions up to 1260x960 pixels using only a 250K CCD. Unless you understand our technology (some of which is proprietary and trade secret) it is easy to disbelieve that this can really be accomplished. After all, if it could be done, Kodak, Polaroid, Sony and others would have done so, right? Wrong.
I'll add here that additional details about Pixera and its technology are available in the January and September issues of Advanced Imaging Magazine in articles written by Mr. Charles Reis...
The Pixera Professional (the camera we sell in microscopy) achieves 4x the resolution of our CCD this way. The key to our high resolution technology is three related pieces. 1.) Our camera head contains an electro-mechanical light refractor (patented in the US and Japan) in front of the CCD. We use this light refractor to shift the incident light from the subject onto the pixel elements. This mechanism is extremely accurate down to the sub-micron level. 2.) We use a proprietary (and patented) color filter pattern for our CCD. It's custom made for Pixera to work with our light refractor. 3.) We have proprietary software image processing software that works in conjunction with the light refractor and CCD filter pattern to achieve the image quality and resolution of our images. Pixera uses 100% software image processing to generate its images, the only image processing done in the camera is AGC and a little Gamma.
When taking a high-res picture (800x600 or above), our camera takes 4 exposures of the subject. This takes a little under a second. For each exposure, our light refractor shifts the incident light in a particular direction for a particular distance in order to get four separate and distinct samples. We do not pixel double or enlarge. To understand this in more depth, look up the patent. Mr. Yuji Ide, founder of Pixera is the principal author. This process is analgous to shifting the CCD for each exposure like some other high priced, high-resolution cameras do.
After each exposure, the raw analog pixel data from the CCD is sent to our interface card where all we do is convert the analog raw pixel data into digital raw pixel data. The raw digital pixel data is then sent into the computer (Winows 95, NT, or Mac) for processing. We do not compress any of the information for transfer into the computer. Another benefit of this technique and technology is that we get two color samples for each pixel. So our camera has the color information and reproduction of a 2-CCD camera. So even though we do have to interpolate to get the 24-bits of color (as do all single CCD cameras) we at least start with twice the information. Our software image processing of course understands our color filter patter, image shifting, etc., and processes the raw pixel information into a image that is approx. 3.6 million pixels.
Our camera also functions as a "video" camera although no formatted video signal is generated. We simply stream the raw pixels (without using the light refractor) into the computer and build the motion frames on the fly. The faster the computer, the faster the video...Today we are hardware limited by our card architecture to 10 fps, next year we won't have this limit.
This is it in a nutshell. We don't pixel double, we don't interpolate other than for color and a little to adjust for aspect ratio, and we don't do "massive pixel enlargement". If anyone wants or need additional information I'd be glad to discuss it with you. If you still don't believe it, we have a couple of thousand customers that did, and we'd like the opportunity to turn you into a believer as well...
} It seems odd to blast Polaroid for introducing } a camera that was $5000 less than their nearest competitor when introduced. } The good news is that prices always go down, and quality goes up.
Gee, I guess then it's really odd to blast Pixera for introducing a camera that is several thousands less than anyone (including Polaroid) simply because you don't believe what the camera is doing and even worse because you don't take the time to understand......
One place where I do agree with Mr. Gruteleuschen is - We at Pixera believe that prices SHOULD always go down and quality go up. That's why we're working hard on new cameras of 2 and 4 million pixels based on our technology. And you can bet that they'll be priced and have quality that some people, at least, will find hard to believe....
I'm not trying to make this personal at all. I'm just tired of "people of science" on various news groups and listservers taking unwarranted and inaccurate potshots at Pixera's products without really knowing the facts.
I appolgize for my unwarranted swipe at commercial institutions. Dr. Connolly has provided an appropriate response--see below. However, his request did not state clearly the conditions and considerations for an exchange of human tissue and since there is considerable controversy regarding these issues amongst pathologists these days I believe this community (Microscopy) should be careful with language as well as any exchange of tissues.
I believe you're thinking of HMDS, hexamethyldisilizane. Most EM supply houses sell it, I think. Procedures for use were covered in a note in the May 97 issue of Microscopy Today, and Scott Whitaker (sp? correct me if I'm wrong) as information on this on the U Florida web pages at: http://www.biotech.ufl.edu/~emcl/tips.html
I've used it successfully. Email me with specifics about what you're up to, and maybe I can give you some ideas.
Phil
} Dear Colleagues: } } I remember there have been reports since many years ago regarding to the } alternative chemical treatment in place of critical point drying using a } critical point dryer. Can any of you tell the name(s) of the chemicals and } the companies which sell them? Or your experience using such chemicals? Sorry } for not being able to look up a SEM book for myself. } } Thanks! } } Yuhui Xu } Core EM Facility } Dana Farber Cancer Institute }
Philip Oshel PO Box 5037 Station A Champaign, IL 61825-5037 (217) 355-1143 oshel-at-ux1.cso.uiuc.edu or poshel-at-hotmail.com ***** looking for a job *****
If anyone is looking into this topic or knows of any relevant references or SEM methods, I would greatly appreciate the information. I have scoured our meager library and the current contents database with virtually no luck. My kingdom for access to the science citation index.
Thank you so much for any aid you can provide.
Jill Craig University of Northern British Columbia
I am still using an old ETEC Autoscan (SEM, vintage 1973). It is still working well after more than 12 000 hrs of usage and usually we get the results that we want.
However, a calcium containing deposit has been forming in the cooling water supply (in Cu tubes, in cooling coils around diff.pump, in heat sinks, ect). The microscope was donated to us several years ago but was not connected for the last 18 months to the recirculating water system in our laboratory ( we are using filtered tap water). The water flow has now been reduced drastically over the last few weeks and I fear that the 'pipes' may eventually totally block up.
What is the best (and safe) way to reduce or remove this deposit. Back-flashing was only partially successful.
Any suggestion ?
Thank You
Hans Brinkies SWINBURNE, University of Technology School of Engineering and Science Electron Microscopy Laboratory HAWTHORN, 3122, Australia
Sorry folks; the Discovery channel has CHANGED the air times of Dennis Kunkel's microscopy, described on the listserver yesterday. Here's the new message:
} Dear Friends and Associates, } Sorry for the incovenience but Discovery Channel has preempted } Movie Magic for this week. It will air the following week (see below). } Discovery Channel's documentary series, "Movie Magic", is airing a } segment called "Far Out Creatures" that features special effects used in } the making of science fiction films (Alien Resurrection and Starship } Troopers). Part of the 1/2 hour segment will include a short interview } with me and some of my "MicroAliens". I thought this might be of } interest to you. } } The segment will run: } December 18, 1997 9:30 - 10:00 PM - West Coast Time (PST) } December 19, 1997 1:30 - 2:00 AM - West Coast Time (PST) } December 20, 1997 2:00 - 2:30 PM - West Coast Time (PST) } } Please check your local listing for the time of Movie Magic on the } Discovery Channel (or check their website schedule at } www.discovery.com/diginets/discovery/discovery.html). } } Best Regards, Dennis Kunkel } } *********************************************** } * Dennis Kunkel Ph.D. * } * Pacific Biomedical Research Center * } * University of Hawaii * } * * } * email - kunkel-at-pbrc.hawaii.edu * } * www - http://www.pbrc.hawaii.edu/~kunkel/ * } **********************************************
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
I have just learned that there has been a delay in the distribution of the
Call for Papers and Advanced Registration for the Microscopy & Microanalysis '98 Meeting in Atlanta, Ga July 12-16
due to a delay at the printers.
It is expected that they will be mailed during the week of Dec. 15th.
In order to avoid having to reply to repeated questions which have started to flow in my direction I have taken the liberty of posting this Email message to both the MSA Membership as well as the Microscopy Listserver Databases.
As additional information becomes available I will make sure that the M&M'98 WWW pages are updated. These WWW pages are directly accessible from the URL
I would like to hear comments or suggestions about sample preparation of biofilms builded up by methanogenic and acidogenic facultative anaerobic bacteria deposited on sand grains for SEM. So far, we have done fixation using glut araldehide and further dehydration with alcohol.
Does anyone have any experience in sampling sand grains from a bioreactor in order to preserve the biofilm for SEM observations?
Thanks in advance,
Nora Pratta Centro Regional de Investigacion y Desarrollo Santa Fe - Argentina
Hi everyone, I would like to hear comments or suggestions about sample preparation of biofilms builded up by methanogenic and acidogenic facultative anaerobic bacteria deposited on sand grains for SEM. So far, we have done fixation using glutaraldehide and further dehydration with alcohol. Does anyone have any experience in sampling sand grains from a bioreactor in order to preserve the biofilm for SEM observations? Thanks in advance, Nora Pratta Centro Regional de Investigacion y Desarrollo Santa Fe - Argentina
You might want to repost this to the biofilms listserver: biofilms-at-net.bio.net. "Conventional" EM (TEM, SEM) has limited utility in study of most bacterial biofilms because they suffer much artifact introduction (spatial distortion) in the drying process. I would encourage you to supplement your SEM studies with ESEM (environmental scanning electron microscopy; what I call "hydrated" SEM), laser confocal microscopy, or cryosectioning light microscopy. I can help you with confocal and can refer you to people who can assist with the rest. Rob Palmer Director - Biofilm Imaging Facility CEB/UT
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
This is the text of Stephen Kohn's message. It was attached as a Word document which makes it a little harder to open.
I wonder if it would be worthwhile to provide some guidance or reminders to the list members as to what makes for good message practice. In addition to these attachments, I have been seeing a number of messages packed with HTML coming across. Most of those appear to come from the Microsoft Outlook mail program. I would rather we stick to plain text so that the most can read the message with the least difficulty.
Regarding the comments made during the past week about the use of inexpensive high-resolution digital cameras for microscopy purposes, please note the following update information about The Pixera "Professional" camera which clarifies any possible misunderstandings about the product that subscribers may have:
Pixera's most recent software upgrade - Version 1.2, contains a 10 frame per second "fast viewfinder" window. This is equivalent to the speed of what many digital camera video conferencing systems run at.
TWAIN compatibility has been improved - Pixera recently tested more than 10 programs from image analysis vendors/providers and found the Pixera Professional camera to be TWAIN compliant with all of them. Feel free to contact me If you'd like a copy of the list.
RGB Color mix in pre-image capture mode now allows for viewing a color balanced image which is equivalent to the final image captured.
With reference to other comments made, the variable lens Pixera Professional "can ship" an uncompressed 3.7 megabyte image to the computer for viewing and or image analysis purposes.
"While I have a vested interest in Pixera as an employee, the purpose of this reply as stated above is to clarify details regarding the system's capability and is presented as a non-commercial message for informational purposes only.
Mike Saulnier is no longer with IVS. Please remove this address from your mailing-list:
msaulnier-at-ivsinc.com
Thanks, Dave Shipman Network Administration
} ---------- } From: Dave Shipman } Sent: Wednesday, December 10, 1997 5:51 AM } To: Dave Shipman } Subject: Notification: Inbound Mail Failure } } The following recipients did not receive the attached mail. Reasons } are listed with each recipient: } } {msaulnier-at-ivsinc.com} msaulnier-at-ivsinc.com } MSEXCH:IMS:IVS, Inc.:SW_DOMAIN:WINNT_1 0 (000C05A6) Unknown } Recipient } } The message that caused this notification was: } } { {Message: Microscopy & Microanalysis '9...} } }
In short, I would like to inquire as to the methods and chemicals that people are using for cleaning their filament assemblys on their electron microscopes. Especially the final step.
When changing our filament for our TEMs, we have in the past cleaned any tarnish or deposits from the filament assembly pieces using a mild polish, then sonicated the parts in a detergergent solution, followed by water rinses, sonication in a few rinses of ETOH, then finally sonication in a freon-based ultra-precision cleaning solution. (The used freon solution is dumped into its own waste bottle.)
We are considering changing this process (especially in consideration of the availability and disposal of the freon solution) and are wondering what other labs use for cleaning their filament assemblys thoroughly. We are especially interested in the final cleaning step (removing any leftover residues).
Thank you for your input. Don't be afraid to be brief.
Gregg Sobocinski Parke-Davis Pharmaceutical Research Ann Arbor, Michigan USA Sobocig-at-aa.wl.com
A related question is: Does anyone know if the proceedings of the 15th Pfefferkorn conference held in May 1996 are ever going to published by Johari et al.
Matt Libera Stevens Institute of Technology
henk-at-vt8200.vetmed.lsu.edu wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello All, } Does anyone know what has happened to Dr. Om Johari and the journal } Scanning Microscopy. I have been unable to contact him via e-mail, } phone, } or fax and have not been able to find reference to the journal in over } two } years. Have he and the journal "retired"? } } Bill Henk } Dept. of Anatomy & Cell Biology } LSU School of Veterinary Medicine } Baton Rouge, LA 70803 } phone -(504)346-3237 } e-mail - henk-at-vt8200.vetmed.lsu.edu
Anyone using HF: Where do you buy your acid neutralizing chemicals? We've purchased the whole HF safety kit in the past, but somehow the chemicals never come out right - I currently have 2 bottles of the HF inactivator (#2) and part of a bottle of the acid neutralizer (#4). The neutralizer is just sodium carbonate (right?) - any reason why I shouldn't just use that from any source? I've spoken with chemistry types from several vendors and nobody is sure. And to buy just the Mallinckrodt stuff packaged for the safety kit is insane - they can't sell the regular jar; I'd have to buy a tub of it. So, I'm hoping someone out there has a wonderful answer - I'm afraid to assume anything with handling this particular chemical.
I'm sure that everyone will tell you that the lifetime of your diamond knife depends on what you cut, who uses it, and how you take care of the knife - it's all true. The lifetime of a knife can be measured in minutes or years - it's up to you. For best results, treat your diamond knife as you would your toothbrush, don't share it with anyone. In truth, a knife needs to be sharpened when the sections it cuts no longer meet your needs. I map the use of my knife edge and systematically work my way across the knife as it degrades with use. For particularly tough samples I may use a previously degraded region of the knife for the initial cuts. With regard to cleaning, minimize cleaning and never allow sections to dry on the knife.
My first diamond knife was a 4 mm knife that lasted 8 years, cutting 4 to 8 hours a week on glass, metals, semiconductors, and dielectrics. So much for predicting lifetimes. Typically there was only myself and a trained tech using the knife. My blocks are made with Spurrs, the blockfaces are prepared on the microtome using a glass knife, and are only 50 to 100 um across. I am meticulous about the preparation of the blockface, the orientation of the sample in the blockface, and strictly minimize the size of the embedded sample that the diamond knife cuts.
Phil Swab Advanced Coatings Division Advanced Refractory Technologies Inc. Buffalo, NY
} ---------- } From: smithde-at-valunet.com[SMTP:smithde-at-valunet.com] } Sent: Tuesday, December 09, 1997 9:10 AM } To: microscopy-at-Sparc5.Microscopy.Com } Subject: Diamond Knife } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } ---------------------------------------------------------------------- } -. } } Can anyone tell me about how often a diamond knife needs to be } sharpened? I } realize it would depend on the amount of wear it gets. Our EM dept. is } only } open three days a week, with cutting being done about 2-4hrs a week. } Our } knife was sharpened six months ago and seems to be getting dull again. } Is } this to be expected? } }
I am also beginning to evaluate hand-held digital cameras for an upcoming image analysis project. The recent discussion has been very helpful for me - thank you to everyone for posting to the general list.
One product I haven't seen mentioned is the Minolta RD-175. Does anyone have experience with this camera, or know how it fits into the spectrum? Does it use the JPEG compression that Dr. John Russ spoke about? Does it provide good color representation? I should mention that I want a camera that is not "tethered to the computer".
Thanks again for your help,
Karen
-- Karen Zaruba kszaruba-at-mmm.com 3M Company, St. Paul, MN 55144 "Opinions above are my own, not necessarily my employer's"
} ... Anyone using HF: } Where do you buy your acid neutralizing chemicals? We've purchased } the whole HF safety kit in the past, but somehow the chemicals never come } out right - I currently have 2 bottles of the HF inactivator (#2) and part } of a bottle of the acid neutralizer (#4). The neutralizer is just sodium } carbonate (right?) - ...
I might suggest only that the neutralizer be Ca carbonate which would allow production of the inert by-product CaF.
cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
ASTIMEX has a set of 53 minerals - standards for microanalysis, in a typical mount of 1 inch diameter. This includes two garnets - pyrope Mg3Al2Si3O12 and almandine Fe3Al2Si3O12.
The address:
ASTIMEX Scientific Ltd. 351 Wellesley Str. East Toronto Canada M4X 1H2 fax: (416) 961-2402 e-mail: jcr-at-quartz.geology.utoronto.ca (Prof. J.C. Rucklidge)
Best regards,
Wojtek xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx Dr Wojciech J. Przybylowicz 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 xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx
Please can you give me the necessary information/instructions how to realize publication of positions of Philips Electron Optics on the MSA's Listserver.
I am Jacques Weterings personnel manager of Philips Electron Optics in Eindhoven the Netherlands.
What are you cutting? Steel? My diamond knives are sharp after 10 years of daily use. We are cutting neural tissue embedded in epon(very soft) We have never resharpened them.
Perhaps your knife was not sharpened well. Or perhaps your knife was beyond repair in the first place.
Sally
On Tue, 9 Dec 1997, Diane M. Smith wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Can anyone tell me about how often a diamond knife needs to be sharpened? I } realize it would depend on the amount of wear it gets. Our EM dept. is only } open three days a week, with cutting being done about 2-4hrs a week. Our } knife was sharpened six months ago and seems to be getting dull again. Is } this to be expected? } } }
At 08:55 AM 12/10/97 +0100, Heinz Fehrenbach wrote: } can anyone of this newsgroup give me information about and subscription } procedure to any related newsgroup ? In particular, I would appreciate } to hear whether there is a special newsgroup discussing immuno-histology, } -fluorescence, -blotting, and pathology subjects.
There is an immunohistochemistry list, called ipox-l
To subscribe, send a message to majordomo-at-pathology.stanford.edu and write subscribe ipox-l in the body of the message. I believe that there is also a pathology list, and instructions for this are available at "The Histotech's Home Page" (http://www.histology.to)
Best regards, Steven E. Slap ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
Om Johari sent out an announcement either late las year, or earlier this year, that he was retiring. He also said that he hadn't found anyone who was willing to take over the job of editing the Scanning Microscopy journal or organizing the annual meeting. I don't have a current email or regular mail address to reach him. It's probably not likely that anything not currently in press will be published by him.
Call Prof. Wil Bigelow in Ann Arbor, 764-3321. I would be surprised if he did not recommend a final rinse/sonication in isopropanol, if freon is a no-no for you. He will certainly have some suggestions for the overall cleaning process also.
Larry
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dr. Lawrence F. Allard Senior Research Staff Member High Temperature Materials Laboratory Oak Ridge National Laboratory 1 Bethel Valley Road Bldg. 4515, MS 6064 PO Box 2008 Oak Ridge, TN 37831-6064
} I wonder if it would be worthwhile to provide some guidance or reminders to } the list members as to what makes for good message practice. In addition to } these attachments, I have been seeing a number of messages packed with HTML } coming across. Most of those appear to come from the Microsoft Outlook mail } program. I would rather we stick to plain text so that the most can read the } message with the least difficulty.
I agree. While I have the capability of downloading and viewing such messages, the bother is such that I will not do so. Let's keep to plain text.
Colleagues... we have old Hitachi S-650 SEM with KEVEX ANALYST 8000 microanalyser with old architecture ( on LST-11 minicomputer). Can anyone give a advice about cheap and easy ways to change this system on modern system on PC platform? What we need to this? Thanks, Yury Shipilov AME Department UofA
I'm in the market for a new inverted microscope that will form the basis of a Fura 2, Ca++ imaging/photometry system. I am considering either a Leica DMIRB or a Nikon TE200/300. Has anyone compared these microscopes and their long working distance Fluor objectives (20, 40 and 63 or 60X)? Any help would be appreciated.
--
William F. Jackson, Ph.D. Professor Department of Biological Sciences 5380 McCracken Hall Western Michigan University Kalamazoo, MI 49008
I have used a diamond knife without any need for resharpening for about 9 years, only cutting plant material. Although I did not cut with it every day, I was the full time technologist in this lab doing most of the ultrathin sections for a variety of researchers. I NEVER cut any thick sections with that knife, and I was careful to always have very small block faces and handle my knife very carefully. If I ever had any sections stick and dry on the knife edge, I would wipe them off with alcohol soaked balsa wood. (first I soaked that balsa in acetone for several weeks to remove all the resin in the wood) Although nobody really recommends wiping the knife edge with anything, I find that it is essential to really have a clean edge. I wipe it under the microscope, and keep wiping until I can see that it is truly clean, but of course I'm careful to avoid using much pressure, or any force perpendicular to the knife edge. BEING GENTLE is the key. If one isn't careful, a lot of garbage can dry on the back of the knife edge without you knowing that it is there. That is why I inspect it carefully under a dissecting microscope, looking especially at the back of the knife edge. This garbage can make you think that your knife needs resharpening, when in fact it is only dirty. In order to loosen up any dirt on the knife edge, I soak it overnight or over the weekend in alcohol. This will not harm the knife, in fact I know a researcher who STORES his diamond knife under alcohol. Of course acetone will ruin it almost instantly, so DON'T EVER MAKE THAT MISTAKE, because the acetone will quite readily dissolve the glue that holds the diamond in place. But the alcohol is perfectly fine.
In a human pathology lab now, I notice that our knives get sharpened more frequently, perhaps after 1 or 2 years, depending on the size of the knife. We cut sections absolutely every day, and very big sections, where a single section might cover a significant portion of the grid. That way the pathologists don't need to waste a lot of time scanning around looking for the section!!
Hope this helps, Garry
} ---------- } From: Murphy, Judy[SMTP:murphy-at-sjdccd.cc.ca.us] } Sent: 9 December, 1997 11:24 } To: Diane M. Smith; MSA } Subject: RE: Diamond Knife } } It also depends on what is being cut as well as who is doing the cutting. } If it is plant material, 6 months is a LONG time. Animal tissue is softer } but again some animal tissues have hard components which are hard on the } knife edge. } The experience of the cutter obviously also comes into play. The less } experience, usually the more resharpenings may be necessary. Are they also } taking thicks on the same knife?
It is also wise to have some Calcium Gluconate 2.5% gel handy to apply liberally to the affected area and cover loosely with gauze wrap. It is available by prescription only I believe.
Honey Nut Cheerios Harry ---------- ----------------------------------------------------------------------- .
Tamara Howard wrote:
} ... Anyone using HF: } Where do you buy your acid neutralizing chemicals? We've purchased } the whole HF safety kit in the past, but somehow the chemicals never come } out right - I currently have 2 bottles of the HF inactivator (#2) and part } of a bottle of the acid neutralizer (#4). The neutralizer is just sodium } carbonate (right?) - ...
I might suggest only that the neutralizer be Ca carbonate which would allow production of the inert by-product CaF.
cheerios, shAf -- {\/} /\ {\/} /\ {\/} /\ {\/} cogito, ergo zZOooOM {\/} /\ {\/} /\ {\/} /\ {\/} Michael Shaffer, R.A. - University of Oregon Electron Probe Facility mshaf-at-oregon.uoregon.edu -or- mshaf-at-darkwing.uoregon.edu http://darkwing.uoregon.edu/~mshaf/
I would like to sincerely apologize to The Pixera Corp. for the = misinformation in my post to the listserv. My intention was not to = disinform. I am not an expert ... just a curious student trying to = learn a little by generating discussion with the previously posted = comparison.
IMAGE RESOLUTION=20 Pixera Their web site states:=20 Resolution in still mode 1.2 meg (1260x960) Image size - 1.3 meg(1280x1024) =20 Polaroid DMC 1.9 meg (1600x1200) Kodak MDS120 1.2 meg (1280x960)
A number of private messages send to me expressed a concern with the = image resolution claims these systems make given the effective = resolutions of the CCDs used.=20
IMAGE TRANSFER I mistakenly stated that Pixera's transfer rate is limited to the = serial port rate. The truth is they have a PCI or PMCIA connection. In = general, is slower image capture is a limitation of these sytems? I am = currently trying to reach tech service for each company to learn more = about how the images are transfered and what the realistic transfer = rates are (basically how long does it take to capture 2 images = consecutively).
Kevin Brent Smith Graduate Student University of Louisville Biology Dept. Life Science Bldg. Rm.#12 Louisville, KY 40292 Phone: (502) 852-6773 Fax: (502) 852-0725
Looking to identify MELANIN in fungal walls. There are no antibodies for it (Its not a protein- I don't think), and I can't find any lectins for it. It is diagnostic in urine samples, and skin, hair etc.
o.k., surely someone somewhere - one of those great old Histologist / microscopists has some eye-of-newt & pinch of bat-wing technique for staining it - possitively would be even better - but we have non-melanized mutants whcih can be used as a control (putatively we can turn melanization On/Off and this is what we are trying to verify, eh?).
Any hints or techniques would be greatly appreciated. Applicable microscopies include SEM (SEI & BEI), TEM, LM, Confocal, and fluorescence. Rather NOT attempt any autoradiography techniques though.
Thank you in advanced for any help!
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"640K ought to be enough for anybody." -- Bill Gates, 1981
Harry - Thanks for pointing this out! Our safety department gets us a fresh tube every year; it is kept in a special pouch on the wall next to the hood where we use and store the HF. We also have an "Instructions for physicians" handout with the gluconate, it deals with treatment following HF exposure.
On Wed, 10 Dec 1997, Ekstrom, Harry wrote:
} It is also wise to have some Calcium Gluconate 2.5% gel handy to apply } liberally to the affected area and cover loosely with gauze wrap. It is } available by prescription only I believe. } } Honey Nut Cheerios } Harry } ----------
We are looking for some Teflon tweezers, to dip a TEM sample in HF. Anyone know of a source?
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University Evanston, IL 60208-3108 tel: (847) 491-3996 fax: (847) 491-7820 email: l-marks-at-nwu.edu http: //www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
I am seeking the advice of any member who wishes to make this a truly joyful season.
I am working with some skin samples, (human, mouse, rat) that are between 100=B5m to 150=B5m thick. They have been labeled with various 1=B0s and three 2=B0s. The labeling is specific and strong. The problem is the clarification of the tissue and the proper mounting in a suitable fashion for confocal. Most of the problems stem from the fact the tissue is so thick; and before you ask, yes it has to be that thick. I am looking for the relationship between matrix, nerve and blood structures. I have performed the 3D analysis successfully on smaller, 40=B5m sections, but clearing the tissue becomes important in the thicker sections for proper imaging.
I have tried dehydration w/ EtOH 70%-100% and Methyl salicylate, but the crystals left behind by the Me-S and washing them displaced the tissue.
I have tried 70-30 v/v glycerol but only moderately improved my image.
I dislike using anything which might leave an organic residue. That includes old favourite cleaners like "Brasso" and "Wenol". AND even the organic solvents which YOU HOPE remove that greasy residue can leave their own residues. Steve Chapman (whom God preserve) advises that "Wenol plus K" in particular leaves behind an organic film intended to retard oxidation.
So my preference is for an aqueous polish which rinses off with copious volumes of our glass distilled water.
Microgrit alumina (say grade 1600) on a damp cotton bud is fine. Or you can buy polishing alumina ready made up (you may have it already) from say Buehler.
But my preference is for diamond paste on a cotton bud. ProScitech sold me our existing tube which was 5g. Its 0.25 micron grit, quality of diamond 100k (whatever that matters). One uses so little it seems to last forever.
Mel Dickson President, Australian Society for Electron Microscopy Director, Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
I dislike using anything which might leave an organic residue. That includes old favourite cleaners like "Brasso" and "Wenol". AND even the organic solvents which YOU HOPE remove that greasy residue can leave their own residues. Steve Chapman (whom God preserve) advises that "Wenol plus K" in particular leaves behind an organic film intended to retard oxidation.
So my preference is for an aqueous polish which rinses off with copious volumes of our glass distilled water.
Microgrit alumina (say grade 1600) on a damp cotton bud is fine. Or you can buy polishing alumina ready made up (you may have it already) from say Buehler.
But my preference is for diamond paste on a cotton bud. ProScitech sold me our existing tube which was 5g. Its 0.25 micron grit, quality of diamond 100k (whatever that matters). One uses so little it seems to last forever.
Mel Dickson President, Australian Society for Electron Microscopy Director, Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
Hi Richard, Our lab studies frog melanophores which contain hundreds of melanosomes (melanin-filled pigment granules). I have had a great deal of success visualizing melanosome distribution on the confocal by imaging backscattered (reflected) light. Using this technique allows me to double stain cells with rhodamine & fluorescein and superimpose reflected light to create very nice 3 color images. On our old Zeiss LSM210 this is possible by scanning with the 543 HeNe laser line (usually used to excite rhodamine) and detecting for green fluorescence (usually used to detect emitted light from fluorescein). You can accomplish the same effect by removing the fluorescent filter block altogether, but images acquired this way are misaligned with respect to fluorescence collected thru the dichroic. The only difficulty I've had using this technique is that the samples must be sufficiently below the plane of the coverslip so that reflection from the glass doesn't contribute to the backscatter image.
Hope you find this input useful. Best,
Steve Rogers Dept. of Cell & Structural Biology & The Beckman Institute - Optical Visualization Facility University of Illinois -at- C/U srogers-at-delphi.beckman.uiuc.edu
} Looking to identify MELANIN in fungal walls. There are no } antibodies for it (Its not a protein- I don't think), and I can't } find any lectins for it. It is diagnostic in urine samples, and } skin, hair etc. } } o.k., surely someone somewhere - one of those great old Histologist } / microscopists has some eye-of-newt & pinch of bat-wing technique } for staining it - possitively would be even better - but we have } non-melanized mutants whcih can be used as a control (putatively we } can turn melanization On/Off and this is what we are trying to } verify, eh?). } } Any hints or techniques would be greatly appreciated. Applicable } microscopies include SEM (SEI & BEI), TEM, LM, Confocal, and } fluorescence. Rather NOT attempt any autoradiography techniques } though. } } Thank you in advanced for any help! } } } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 352 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } } } "640K ought to be enough for anybody." } -- Bill Gates, 1981
Its fair and reasonable for Steven Slap to provide this information, but why does he not indicate that he is the editor of the suggested histology site?
Sure his name appears on that site and his company is shown as a sponsor (a cheap form of advertising), but no connection is made and most visitors would not realise that Steven Slap is the manager of the sponsoring firm and thus has a commercial interest: In essence it is a commercial site. Predictable his company is the only consumable supplier company shown. Nothing wrong with his sponsoring or editing such a page but visitors to the page and readers of this listserver are entitled to know not just who is sponsoring what but what commercial interests the editor has. Its known as propriety! Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au
---------- } From: Energy Beam Sciences, Inc. {ebs-at-ebsciences.com} } Dear fellow microscopists, } } At 08:55 AM 12/10/97 +0100, Heinz Fehrenbach wrote: } } can anyone of this newsgroup give me information about and subscription } } procedure to any related newsgroup ? In particular, I would appreciate } } to hear whether there is a special newsgroup discussing immuno-histology, } } -fluorescence, -blotting, and pathology subjects. } } There is an immunohistochemistry list, called ipox-l } } To subscribe, send a message to majordomo-at-pathology.stanford.edu and write } subscribe ipox-l in the body of the message. I believe that there is also a } pathology list, and instructions for this are available at "The Histotech's } Home Page" (http://www.histology.to) } } Best regards, } Steven E. Slap } ******************************** } Energy Beam Sciences, Inc. } Adding Brilliance To Your Vision } ebs-at-ebsciences.com } http://www.ebsciences.com/ } ******************************** }
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Subject: Time:5:39 PM OFFICE MEMO RE} Cooling water problems Date:12/10/97
Hans Brinkies wrote, "I am still using an old ETEC Autoscan (SEM, vintage 1973). It is still working well after more than 12 000 hrs of usage and usually we get the results that we want.
"However, a calcium containing deposit has been forming in the cooling water supply (in Cu tubes, in cooling coils around diff.pump, in heat sinks, ect). The microscope was donated to us several years ago but was not connected for the last 18 months to the recirculating water system in our laboratory ( we are using filtered tap water). The water flow has now been reduced drastically over the last few weeks and I fear that the 'pipes' may eventually totally block up.
"What is the best (and safe) way to reduce or remove this deposit. Back-flashing was only partially successful.
"Any suggestion ?
"Thank You
"Hans Brinkies SWINBURNE, University of Technology School of Engineering and Science Electron Microscopy Laboratory HAWTHORN, 3122, Australia" **************************************** Hans, The same problem occurs in lots of water-cooled, high power equipment such as vacuum tube amplifiers, x-ray tubes, and particle accelerators. For components with copper cooling water passages we use only low conductivity water (min of 750K-ohm-cm: 1 Megohm-cm is better) in a closed-loop cooling system. Even then deposits will accumulate. We use water flow switches to ensure at least a minimum water flow in sensitive equipment, and try to keep an eye on the pressure drop across each water cooling circuit, back-flushing with a dilute solution of Sulfamic acid when we see a clear rise in the pressure drop at the required flow. Hope this helps keep your ETEC Autoscan running. Dennis Collins DGCollins-at-lbl.gov
Dear Yuri, I have been researching the same thing to replace my two old Kevex 8000s. So far I have found iXRF, (www.ixrfsystems.com) who specialize in putting new computers on old Kevex's, ANS, (www.qtmsys.com) who have a very inexpensive product, and, of course, Kevex has an upgrade that is a little more. All are worth talking to. You wrote:
} Colleagues... } we have old Hitachi S-650 SEM with KEVEX ANALYST 8000 microanalyser with } old architecture ( on LST-11 minicomputer). Can anyone give a advice } about cheap and easy ways to change this system on modern system on PC } platform? What we need to this? } Thanks, } Yury Shipilov } AME Department } UofA } 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 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
Dear Sara, We have been running the VCR Ion Mill for about eight years, now. We have found the instrument to be solid, reliable and easy to use and clean. The people at VCR are very helpful and easy to deal with. Ours is an older model, now they have one run totally by computer. You wrote: } Hi All } } We need to buy a new ion mill for our lab. Any suggestions, warnings, } tips would be appreciated. } } Thanx in advance } } Sara Prins
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 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
Dear Gregg, Everyone has their favorite technique and all will work. I use a good scrub with Wenol, a rinse in acetone, which removes any Wenol residue, a check under 20X dissceting scope to be sure I got it all clean, a rinse in denatured ehtanol, then a final rinse in pure ethanol and dry with hot air. The hot air dry is important to prevent drying residue. Each time I say "rinse", I mean immersing the assembly in the solvent and sonicating for at least a minute. Another technique that helps is to sonicate the piece first in a 10% oxalic acid solution, then rinse in several distilled water changes. Dry with clean alcohol as above. The oxalic acid loosens the residue and saves hard scrubbing. Good luck. You wrote: } } In short, I would like to inquire as to the methods and chemicals that people } are using for cleaning their filament assemblys on their electron microscopes. } Especially the final step. } } When changing our filament for our TEMs, we have in the past cleaned any } tarnish or deposits from the filament assembly pieces using a mild polish, then } sonicated the parts in a detergergent solution, followed by water rinses, } sonication in a few rinses of ETOH, then finally sonication in a freon-based } ultra-precision cleaning solution. (The used freon solution is dumped into its } own waste bottle.) } } We are considering changing this process (especially in consideration of the } availability and disposal of the freon solution) and are wondering what other } labs use for cleaning their filament assemblys thoroughly. We are especially } interested in the final cleaning step (removing any leftover residues). } } Thank you for your input. Don't be afraid to be brief. } } Gregg Sobocinski
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 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
Dear Laurence, A forceps supplier such as Fine Science Tools may have them, but a cheaper source is the Optician that handles soft contact lenses. That's where I got mine. Some drup stores have them with contact lens supplies. They may just be plastic, but should stand HF. You wrote: } } We are looking for some Teflon tweezers, to } dip a TEM sample in HF. Anyone know of a source? } } ++++++++++++++++++++++++++++++++++++++++++++++++ } Laurence Marks } Department of Materials Science and Engineering
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 fax: 604-822-3619 e-mail: mager-at-interchg.ubc.ca
At 03:44 PM 12/10/97 +0200, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
Dear Colleague,
I think that one should consider the following products:
1) VCR Group XLA 2000 Ion/Atom Mill (USA) Suite 31, 250 East Grand Avenue, South San Francisco, CA 84080
2) Gatan Model 691 Precision Ion Polishing System PIPS (USA) Gatan Inc. 6678 Owens Drive, Pleasanton, CA 94588-3334, USA
3) Bal-Tec RES 100 (Lichenstein) BAL-TEC AG, F=F6hrenweg 16, Postfach 62, FL-9496 Balzers, F=FCrstent= um Lichenstein
4) Technoorg-Linda IV3 or IV3/H/L (Hungary) H-1077 Budapest, R=F3zsa u. 24, Hungary, phone: 36-1-3428-713, fax:= 36-1-3224-089
5) Fishione Instruments Model 3000 Ion Mill (USA) E. A. Fishione Instruments Inc. 9003 Corporate Circle, Export, PA 15632= USA
I think that IonTec (UK) is not present on this market now, but I am not sure. I can give you further details separately if you want. Best wishes, Bela Pecz 11th Dec. 1997 ----------------------------------------- Dr. Bela Pecz Research Institute for Technical Physics H-1325 Budapest, POBox 76 Hungary phone: 36-1-2332 865 fax: 36-1-2332-794 E-Mail: pecz-at-mufi.hu -----------------------------------------
Laurence Marks wrote: ======================================================= We are looking for some Teflon tweezers, to dip a TEM sample in HF. Anyone know of a source? ======================================================= If what you want to dip is something more substantial in size than a TEM grid, then the molded Teflon (R) tweezers found on our website will hold up extremely well. They were originally developed for use in the electronics industry where HF dipping is a pretty frequent and routine operation. But don't try to pick up with them a TEM grid, it won't work.
If it is a TEM grid or grid sized sample you want to pick up, then you will need Teflon coated high precision tweezers which can also be found on our website. It is a constant battle of trade offs: You want the tips to retain their sharpness, therefore the coating must be extremely thin, but at that coating thickness, there is a good chance for some small population of pin-holes to form at the tips. So the protection is not perfect, but it is lightyears better than using unprotected tweezers. Also, Teflon is soft and lacks abrasion resistance and it will, therefore, wear off, some might say too quickly. But the good news is that the tweezers can still be used as ordinary tweezers for other purposes.
Now while the molded Teflon tweezers are a pretty commonly found item, high precision Teflon coated tweezers are not so easily found. My comments relative to pin-holes and the coating being worn off were directed specifically to the SPI brand of this kind of product since we have not made comparisons between our product and anyone else's.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ =================================================
Om's last Scanning Microscopy International meeting was billed as just that last May in Chicago, I was tthere. He has oganised them for 30 years and I guess has done it enough. Some of the groups were talking of continuing their own little specialist gatherings elsewhere in future, who knows?
There is a Pfeffercorn meeting being organised currently:
_______________________________________________- Scanning Microscopy International Sixteenth Pfefferkorn Conference Optimising Scanning Electron Microscopy April 5 - 8 1999 Aberystwyth, Wales, UK
Full details of the conference, updated regularly, can be viewed on the World Wide Web page: http://www.aber.ac.uk/~dbswww/pfeffer16/home.html
Dr Iolo ap Gwynn, (iag-at-aber.ac.uk) Institute of Biological Sciences, The University of Wales, Aberystwyth, Wales, SY23 3DA, UK Fax: +44 1970 62 23 50 ________________________________________________
I am trusting that the journal is continuing because I received a manuscript back this week, along with the reviewers' comments. More work!
Just to join in on a discussion that involves a large part of our daily = tasks, I would like to point out the dangers in cleaning in solvents. If you clean your own E.M., then usually you are also aware of the = "freshness" and source of the chemicals/solvents in your lab. In our = travels we often find that we cannot get the solvents we would like or = need and have to make do with a lot of strange variations. We have also = found contaminated solvents which can cause all sorts of problems. = Usually you can visibly see the residue. It appears as white stains. = It's the invisible residues that catch us some times. The basic rules we follow is as follows: Clean only if necessary. If you find that the test specimen is proving = that the E.M. is not up to spec, only then clean the column bits.=20 If the gun assembly is still fairly clean, do not polish. You will only = add dirt. Use a polishing paste that you know works and what it takes to remove = it. This can only be tested by experience unfortunatly. Pastes we use = vary from Wenol, Pol metal polish, pikal metal polish( supplied with = most Japanese E.M.'s) and Hyprez diamond pastes. All of which work in = different circumstances. As we are normally charging per hour, the = customers don't appreciate us trying to polish away for hours on end. So = we find the diamod pastes work better on the gun parts as they remove = the tungsten very quickly. We then commonly use Ethanol as a solvent for these pastes, as acetone = is not always available. One of the better solvents we do use is = Arklone. But it is also not always available and is very expensive.
The secrets we have learnt are simple and work well. Polish the parts well with cotton buds and the paste. Once it is clean, = immerse them immediately in the ethanol/solvent. This ensures that the = paste does not harden on the parts. Ultrasonically clean the parts for at least 10 minutes a time. Remove = the solvent and fill the beaker again with new solvent and ultrasonic = again. If Arklone is available, then use this as the final solvent. When taking the parts out of the solvent, "polish" again with clean = "solvent dunked" cotton buds. Normally you find that there is still = quite a lot of "dirt" that comes off. Use clean solvent to dunk the buds = into. Beware of fibres that can be left after polishing. Check the parts under a light microscope for any particles, residues or = fibres. Once the E.M. is re-assembled allow it to pump down over night, at = least, before checking for a beam.
With the gun assembly, we would again suggest you do not clean if it is = not necessary, as a light coating of tungsten does not affect = performance that badly. For the higher KV. TEMs obviously this would = have to be monitored very closely.=20 We have had good success in simply ultrasonically cleaning with the = Protrain formula of 10% Silvo, 10% ammonia and water. Rinsing thoroughly = afterwards in water then some alcohol. Again I will point out that we donot always have all the fancy chemicals = most labs have available to them, but if this method works for us, I am = sure the added steps and specialised chemicals listed can only improve = cleaning by a smaller margin.
Good luck and happy polishing.
Luc Harmsen Anaspec, South Africa Technical support for E.M. clients, world wide. Anaspec-at-icon.co.za Tel:+27 (0) 11 476 3455 Fax:+27 (0) 11 476 7290
Can somebody please send a copy of a method how to make holey carbon to me? I know it was on the listserver a while ago but somehow I can't find the e-mail or my printout. Thanx
Are you thinking about having a garage, tag, or yard sale within the next 12 months? Are you planning on going to a flea market?
Will it be fun or a big pain...organized or chaotic? Will you be encouraged or disappointed? And most of all... Will you make enough MONEY to make it worth your time? Start planning now!
Making Money At GARAGE SALES SWAP MEETS FLEA MARKETS (An Insider's Guide) Is your resource to make it happen BIG!
On almost any given weekend across the country, someone is holding a garage sale, attending a swap meet or setting up a booth at a flea market. The quote "someone’s junk is someone else’s treasure" makes these events both plausible and plentiful.
GARAGE SALE KNOW-HOW Turn Your Trash Into Easy Cash! Make hundreds more by Garage Selling smarter, not harder.
Learn insider secrets in our new guide.
This information was gathered from the experiences and expertise of many Garage Sale professionals. Definitely a best of the best. No slick salesman's talk. Just an easy-to-read information guide, packed with the latest techniques to help you put cash i n your pockets.
Here are just a few of the techniques covered in-depth in the guide:
Top selling Garage Sale items. Pricing (Or... "What do I charge for this stuff?!") Advertising- the most effective ways to get customers to come. Set Up & Display- the best methods that will attract buyers and get them to purchase. Pitfalls to Avoid and simple mistakes that will lose you money and ruin your sale. How to create Flyers that get customers to come. Security and other safeguards you should follow. How to cash in on your Antiques and Collectibles. How to create winning signs by using psychological principles. How to determine the length of your sale and which days are best. What garage layout will improve sales. Understanding how to transform your trash into treasure. Plus techniques to boost your sales.
...And Much, Much More!! (OVER 25 PAGES)
Consider the price of this guide to be an investment in your future. It will easily pay for itself after you read it.
These proven methods really can TRIPLE your garage sale income!!!
Here is what a few readers had to say:
"I ordered your guide several months ago. It was extremely helpful. I had a two day sale and made over $500 on stuff from around my home! Your guide was a big help in organizing my sale. Thanks!" (Janice J., Clearwater, Florida)
"I did it! Your guide helped me do what I wasn't really comfortable doing, running my sale. I had the best sale possible. I did so well I'm planning another in a few weeks! It was so busy I could hardly take the money fast enough. Thanks for your help. BTW- I made over one thousand dollars!" (Bill S., Long Beach, California)
"I have your guide, and it is well written and very informative. I appreciate your promptness in delivery. I will surely implement your techniques in my next sale. Thanks again!" (Gregg G., Oak Hill, VA)
Let the guide show you step-by-step exactly what you need to do to have a wildly successful garage sale or yard sale. It will lead you from the very beginning of your preparations... to the grand finale of taking your money to the bank!
This guide is not selling for $20, $30 or $50 like some. I'm willing to give you valuable information at an incredibly low price! In fact, for approximately the cost of feeding your family a fast-food meal, you can discover the "secrets" of making your ne xt garage sale or yard sale hugely profitable.
To order your informative guide, send a check or money order for
only $5.00 if delivered by E-mail (Sent in an easy to use format (RTF) useable by all word processors, even Notepad)
only $10.00 if delivered by US mail (shipping included)
Or, you can order with your VISA or MasterCard!
(Sorry...No checks accepted from outside of the U.S. Only VISA or MasterCard!) (If you live in Canada. Please add $1.00 extra for shipping is non E-mail)
You can order by VISA/MasterCard by calling:
1-703-912-1948 (24 hours a day) (Credit Card Orders Only!)
%%%%% BONUS %%%%% IF YOU ORDER BEFORE DECEMBER 20th I WILL ALSO SEND YOU A COMPILATION OF GARAGE TIPS, TRICKS AND IDEAS (IT TOOK ME OVER 15 HOURS OF ONLINE RESEARCH AND YEARS OF ATTENDING FLEA MARKETS AND GOING TO AND HOSTING GARAGE SALES TO PULL THIS TOGETHER)
***If you would like- print this out on your printer and use it as an order form!*** OR E-mail it to me with your information (addresses below)
Zip Code_________________Country______________________
Internet E-mail Address________________________________ (Your E-mail Address will not be used for any other purpose other than to contact you about your order or send your guide.)
2.I am enclosing:
___ a Check Delivery: E-mail: ___ $5.00 US Mail:___ $10.00
___ a Money Order Delivery: E-mail: ___ $5.00 US Mail:___ $10.00
Please make payments payable to: Summit
3.I want to order by credit card:
Card Type: VISA___ MasterCard___
Card Number:___________________________
Card Expiration Date:__________________
Cardholders Name:______________________
Cardholders Address (if different from shipping address above):
Summit Attn: Guide 2579 John Milton Drive Ste. 105-102 Herndon, VA 20171
OR Send E-mail to: (for Credit Card Orders) garage2-at-hotmail.com or garage2-at-mailexcite.com
**if the E-mail addresses are returned as undeliverable please call us and place a Credit Card order or send the order form in the mail**
Thank You for your order! We'll contact you by E-mail as soon as we have received and shipped your Information Package.
-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-=-= If you wish to be removed from this list place REMOVE in the subject heading and reply to the reply address.
Dehydrate with graded ethanol 50 through 100 % then clear in 2 changes of Xylene 5-10 minutes each, that shoul render the tissues translucent.
-- Begin original message --
} Hello: } } I am seeking the advice of any member who wishes to make this a truly } joyful season. } } I am working with some skin samples, (human, mouse, rat) that are } between 100=B5m to 150=B5m thick. They have been labeled with various 1=B0s } and three 2=B0s. The labeling is specific and strong. The problem is the } clarification of the tissue and the proper mounting in a suitable } fashion for confocal. Most of the problems stem from the fact the } tissue is so thick; and before you ask, yes it has to be that thick. I } am looking for the relationship between matrix, nerve and blood } structures. I have performed the 3D analysis successfully on smaller, } 40=B5m sections, but clearing the tissue becomes important in the thicker } sections for proper imaging. } } I have tried dehydration w/ EtOH 70%-100% and Methyl salicylate, but the } crystals left behind by the Me-S and washing them displaced the tissue. } } I have tried 70-30 v/v glycerol but only moderately improved my image. } } Please help } } Ramin Rahbari } 313-998-3383 -- End original message --
regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
**I'm willing to make the mistakes if someone else is willing to learn from them**
I have not seen any cheap conversions, unless perhaps you do the whole thing yourself (and if your "rates" are very, very cheap). There are a number of
vendors who offer pc conversion packages, but the cost is from about $8000 to $20,000 (US).
Woody White Mcdermott Technology, Inc.
Work http://www.mtiresearch.com
Me http://www.geocities.com/capecanaveral/3722 ______________________________ Reply Separator _________________________________
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Colleagues... we have old Hitachi S-650 SEM with KEVEX ANALYST 8000 microanalyser with old architecture ( on LST-11 minicomputer). Can anyone give a advice about cheap and easy ways to change this system on modern system on PC platform? What we need to this? Thanks, Yury Shipilov AME Department UofA
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We are looking for some Teflon tweezers, to dip a TEM sample in HF. Anyone know of a source?
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University Evanston, IL 60208-3108 tel: (847) 491-3996 fax: (847) 491-7820 email: l-marks-at-nwu.edu http: //www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
Jim Darley wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Its fair and reasonable for Steven Slap to provide this information, but } why } does he not indicate that he is the editor of the suggested histology site? } } Sure his name appears on that site and his company is shown as a sponsor (a } cheap form of advertising), but no connection is made and most visitors } would not realise that Steven Slap is the manager of the sponsoring firm } and } thus has a commercial interest: In essence it is a commercial site. } Predictable his company is the only consumable supplier company shown. } Nothing wrong with his sponsoring or editing such a page but visitors to } the page and readers of this listserver are entitled to know not just who } is sponsoring what but what commercial interests the editor has. Its known } as propriety! } Jim Darley } } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Phone +61 77 740 370 Fax: +61 77 892 313 } Great microscopy catalogue, 500 Links, MSDS, User Notes } ************************ http://www.proscitech.com.au } } ---------- } } From: Energy Beam Sciences, Inc. {ebs-at-ebsciences.com} } } Dear fellow microscopists, } } } } At 08:55 AM 12/10/97 +0100, Heinz Fehrenbach wrote: } } } can anyone of this newsgroup give me information about and subscription } } } procedure to any related newsgroup ? In particular, I would appreciate } } } to hear whether there is a special newsgroup discussing } immuno-histology, } } } -fluorescence, -blotting, and pathology subjects. } } } } There is an immunohistochemistry list, called ipox-l } } } } To subscribe, send a message to majordomo-at-pathology.stanford.edu and } write } } subscribe ipox-l in the body of the message. I believe that there is } also a } } pathology list, and instructions for this are available at "The } Histotech's } } Home Page" (http://www.histology.to) } } } } Best regards, } } Steven E. Slap } } ******************************** } } Energy Beam Sciences, Inc. } } Adding Brilliance To Your Vision } } ebs-at-ebsciences.com } } http://www.ebsciences.com/ } } ******************************** } }
Jim, Don't be so childish. If your company had the gumption and interest to support the profession you serve, perhaps you would have had the insight to establish such a site as Steven has. Ronnie Houston Dallas, TX
} } Does anybody out there } } Looking to identify MELANIN in fungal walls. There are no } antibodies for it (Its not a protein- I don't think), and I can't } find any lectins for it. It is diagnostic in urine samples, and } skin, hair etc. } } o.k., surely someone somewhere - one of those great old Histologist } / microscopists has some eye-of-newt & pinch of bat-wing technique } for staining it - possitively would be even better - but we have } non-melanized mutants whcih can be used as a control (putatively we } can turn melanization On/Off and this is what we are trying to } verify, eh?). } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 352 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu -- End original message -- Richard,
While not quite eye of newt and bat wing, though it doesn't miss that era by much, try the following procedure by Masson (1928) which is still in use today.
Masson-Fontana method for melanin:
Fontana Silver Solution:
10 percent silver nitrate 20 ml Ammonia Distilled water 20 ml
To the 10% silver nitrate add the ammonia a drop at a time untill only a faint opalescence remains. Add the distilled water and filter, allow to stand overnight prior to use. Store in the dark and the solution will be good for about 30 days. Do not reuse.
Method:
1-hydrate the tissue sections 2-wash well in distilled water 3-transfer to Fontana silver solution for 12-16 hours in the dark in a covered container 4-Wash well in 2-3 changes of distilled water 5- (optional step)tone in 1% gold chloride 6-stabelize in 5% sodium thiosulfate for 5 minutes 7-wash in tap water for 5 minutes 8-counterstain wit 1% neutral red for 2 minutes 9-dehydrate, clear and coverslip
NOTE!!!!!Ammoniacal silver decomposes into an explosive compound after time (months)
From Histopathological Stains and their Diagnostic Uses, J.D. Bancroft & A. Stevens
If I can be of further assistance please feel free to call me.
regards, Bob Robert Schoonhoven Laboratory of Molecular Carcinogenesis and Mutagenesis Dept. of Environmental Sciences and Engineering University of North Carolina CB#7400 Chapel Hill, NC 27599 Phone office 919-966-6343 Lab 919-966-6140 Fax 919-966-6123
**I'm willing to make the mistakes if someone else is willing to learn from them**
A coworker needs to prepare plan view and cross sectional TEM samples of metal thin films (30-500 angstroms) on GaAs. Any suggestions on the best sample prep for this type of material? TIA
Dick Fonda
_____________________________________________________________ Richard W. Fonda Naval Research Laboratory (202) 767-2622 Code 6324 (202) 767-2623 fax Washington DC 20375 _____________________________________________________________
The journal Pigment Cell Research has much to say about melanin in it's different forms. Also, work done in our lab some time ago used a whole list of nonradioactive methods for detecting melanin. Check out "Pigmented cells of the stria vascualris and spiral ligament of the chinchilla" by C.G. Wright and D.H. Lee, Acta Otolaryngologica (Stockholm) 108:190-200, 1989 or "Pigmented epithelial cells of the mebranous cassular wall of the chinchilla" by C.G. Wright and D.H. Lee, Acta Otolaryngologica (Stockholm) 102:438-449. There is even a way to see if your nonpigmented cells might actually be producing the precursors of melanin without making the final product. I didn't do any of the work myself- so I can't give you much detail. Hope this info is helpfull.
Karen Pawlowski Sr. Res. Assoc. Dept. of Otolaryngology UT Southwestern Med. Ctr. Dallas, TX
On Wed, 10 Dec 1997 edelmare-at-casmail.muohio.edu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Does anybody out there } } Looking to identify MELANIN in fungal walls. There are no } antibodies for it (Its not a protein- I don't think), and I can't } find any lectins for it. It is diagnostic in urine samples, and } skin, hair etc. } } o.k., surely someone somewhere - one of those great old Histologist } / microscopists has some eye-of-newt & pinch of bat-wing technique } for staining it - possitively would be even better - but we have } non-melanized mutants whcih can be used as a control (putatively we } can turn melanization On/Off and this is what we are trying to } verify, eh?). } } Any hints or techniques would be greatly appreciated. Applicable } microscopies include SEM (SEI & BEI), TEM, LM, Confocal, and } fluorescence. Rather NOT attempt any autoradiography techniques } though. } } Thank you in advanced for any help! } } } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 352 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } } } "640K ought to be enough for anybody." } -- Bill Gates, 1981 }
For the record, the Polaroid PDC-2000 is available for use either "tethered" or "untethered", the latter with onboard storage for 40 or 60 images depending on the model. You will then need to connect to the computer to download your images, though. Price of the PDC-2000 is now under $2K for all models.
Hope this helps your information gathering process. For more, get back to me or check www.polaroid.com
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am also beginning to evaluate hand-held digital cameras for an upcoming image analysis project. The recent discussion has been very helpful for me - thank you to everyone for posting to the general list.
One product I haven't seen mentioned is the Minolta RD-175. Does anyone have experience with this camera, or know how it fits into the spectrum? Does it use the JPEG compression that Dr. John Russ spoke about? Does it provide good color representation? I should mention that I want a camera that is not "tethered to the computer".
Thanks again for your help,
Karen
-- Karen Zaruba kszaruba-at-mmm.com 3M Company, St. Paul, MN 55144 "Opinions above are my own, not necessarily my employer's"
For the record, the Polaroid PDC-2000 is available for use either "tethered" or "untethered", the latter with onboard storage for 40 or 60 images depending on the model. You will then need to connect to the computer to download your images, though. Price of the PDC-2000 is now under $2K for all models.
Hope this helps your information gathering process. For more, get back to me or check www.polaroid.com
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am also beginning to evaluate hand-held digital cameras for an upcoming image analysis project. The recent discussion has been very helpful for me - thank you to everyone for posting to the general list.
One product I haven't seen mentioned is the Minolta RD-175. Does anyone have experience with this camera, or know how it fits into the spectrum? Does it use the JPEG compression that Dr. John Russ spoke about? Does it provide good color representation? I should mention that I want a camera that is not "tethered to the computer".
Thanks again for your help,
Karen
-- Karen Zaruba kszaruba-at-mmm.com 3M Company, St. Paul, MN 55144 "Opinions above are my own, not necessarily my employer's"
The absolute best, fastest, and cheapest way to prepare non-site specific cross section samples with this type and thickness of metallization on GaAs is with the small angle cleavage technique. There is a detailed pictorial outline of the technique in the MRS Specimen Prep for TEM of Materials IV, Vol 480 that just came out. The authors are myself and John McCaffrey who developed the technique and has several pubs out on it. The technique requires very little in terms of equipment that is not usually found in a lab. Southbay Technology is selling a starter kit that has all the required supplies. I can typically prepare about 9 samples in about an hour and exchange and examine a sample within about 5 minutes to see if it is good. A big advantage to this method is that there is no need to worry about contamination of the sample. I've used this on GaAs and other materials and examined the samples in field emission microscopes with no problems. I do use electronic grade acetone and double rinse them.
For the plan view samples, you will have to dimple and ion mill because of the metallization. Don't forget to protect the good side from ion sputtered material.
If you didn't have the coating and just wanted to make plan view samples from the GaAs, I'd recommend chemically polishing the GaAs from the backside after mechanically thinning to about 100 um to make the plan view samples. Peter Goodhew showed me an inexpensive way to do this. I think that the solution used was 5% bromine in ethyl alcohol (it might have been methanol) which was dripped onto the sample from a burette. The sample was low temperature waxed to a coverslip slide that was put onto a Teflon pedestal with a small amount of vacuum grease mounted in a plastic cup. The grease was not exposed to the solution because it was in the middle of the coverslip and far from the edges. The cup was tilted at an angle of about 30 degrees and was rotating with the use of a small DC motor. A stereomicroscope was used to help determine when the sample was perforated.
You could use this method instead of dimpling if you stopped the process before perforation and then continued the thinning process with ion milling. The disadvantage is that you will not know the thickness left to ion mill.
I hope this helps.
-Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of S.D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
A coworker needs to prepare plan view and cross sectional TEM samples of metal thin films (30-500 angstroms) on GaAs. Any suggestions on the best sample prep for this type of material? TIA
Dick Fonda
_____________________________________________________________ Richard W. Fonda Naval Research Laboratory (202) 767-2622 Code 6324 (202) 767-2623 fax Washington DC 20375 _____________________________________________________________
Sorry, folks; the schedule that I posted last week has been changed. CS } ************************************************************************ } Dear Friends and Associates, } Sorry for the incovenience but Discovery Channel has preempted } Movie Magic for this week. It will air the following week (see below). } Discovery Channel's documentary series, "Movie Magic", is airing a } segment called "Far Out Creatures" that features special effects used in } the making of science fiction films (Alien Resurrection and Starship } Troopers). Part of the 1/2 hour segment will include a short interview } with me and some of my "MicroAliens". I thought this might be of } interest to you. } } The segment will run: } December 18, 1997 9:30 - 10:00 PM - West Coast Time (PST) } December 19, 1997 1:30 - 2:00 AM - West Coast Time (PST) } December 20, 1997 2:00 - 2:30 PM - West Coast Time (PST) } } Please check your local listing for the time of Movie Magic on the } Discovery Channel (or check their website schedule at } www.discovery.com/diginets/discovery/discovery.html). } } Best Regards, Dennis Kunkel } } *********************************************** } * Dennis Kunkel Ph.D. * } * Pacific Biomedical Research Center * } * University of Hawaii * } * * } * email - kunkel-at-pbrc.hawaii.edu * } * www - http://www.pbrc.hawaii.edu/~kunkel/ * } ***********************************************
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.MSA.microscopy.com/ProjectMICRO/Books.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/
} Dear all, } } A coworker needs to prepare plan view and cross sectional TEM samples of } metal thin films (30-500 angstroms) on GaAs. Any suggestions on the best } sample prep for this type of material? TIA } } Dick Fonda
Ion milling of course. Bela Pecz 11th Dec. 1997 ----------------------------------------- Dr. Bela Pecz Research Institute for Technical Physics H-1325 Budapest, POBox 76 Hungary phone: 36-1-2332 865 fax: 36-1-2332-794 E-Mail: pecz-at-mufi.hu -----------------------------------------
Right, prices seem to be dropping. I recently visited one web site = advertising the PDC-2000 for ~$2500 only to visit again a few weeks = later to see it reduced to ~$1600. I haven't seen a comensurate price = drop in the DMC 2000 but that is probably because I haven't recieved a = recent quote. =20 Kevin Brent Smith University of Louisville Biology Dept.
-----Original Message-----
The PDC 2000/40 has a new(45 days old) list price of $1699 and the PDC/2000/60 has a list price of $1999. There has not been a price reduction on the DMC as of yet.
John D. Warren Area Sales Manager Digital Products Polaroid Corporation "See What Develops" 4525 Leonard Parkway Richmond, Virginia 23221-1809 804 254 1011 804 254 1013 Fax warrenj1-at-polaroid.com
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Right, prices seem to be dropping. I recently visited one web site advertising the PDC-2000 for ~$2500 only to visit again a few weeks later to see it reduced to ~$1600. I haven't seen a comensurate price drop in the DMC 2000 but that is probably because I haven't recieved a recent quote. Kevin Brent Smith University of Louisville Biology Dept.
-----Original Message-----
For the record, the Polaroid PDC-2000 is available for use either "tethered" or "untethered", the latter with onboard storage for 40 or 60 images depending on the model. You will then need to connect to the computer to download your images, though. Price of the PDC-2000 is now under $2K for all models.
Hope this helps your information gathering process. For more, get back to me or check www.polaroid.com
} Jim, } Don't be so childish. If your company had the gumption and interest to } support the profession you serve, perhaps you would have had the insight } to establish such a site as Steven has. } Ronnie Houston } Dallas, TX
Fair comment, but as one who visits it occasionally, I'd like to point out that Jim does host an interesting and useful website.
Ritchie
Ritchie Sims phone: 64 9 3737599 ext 7713 Department of Geology fax: 64 9 3737435 University of Auckland Private Bag 92019 Auckland New Zealand
Tom Phillips wrote, I am having a problem with my Bio-Rad MRC-600 and I was wondering if any other users have had a similar one. On the Normal scan speed, I have a very consistent "vibration" in the image in which each horizontal line on the monitor are slightly zig-zagged.
This problem can be caused by 2 problems, and I have experienced both. One is laser instability, and the second is dirty contacts between the detector and the digitizer. I suspect that your problem is the later from what you describe. To fix it remove the front panel of the scan head and disconnect the 9 pin connect coming from the detectors. They are located on the left side of the box, if the scan head is on an upright scope. They are the same as the cable running from the scan head to the computer scan card. Remove them and clean the pins with an eraser to remove the carbon build up that occurs, and then blow out the dust. Electrical contact cleaner or small amount of methanol or acetone should work as well. I do this on a semi-annual basis. If this appears at the 1 second scan speed it will eventually show up in the slow, 4 second scan speed.
Joe Goodhouse Confocal Core Facility Molecular Biology Princeton University
We have a Gatan wide-angle CCD camera with digital processor for EM201 and have been asked to set up an image capture, processing and archiving computer system. I know there are several expensive packages out there but feel that since this is a standard TV signal, 640x480, there should be some effective products,(i.e. All-In-Wonder from ATI) that work together and are available from computer shops.
Does anyone have a similar system set up already (not necessarily for TEM) that uses universal file formats and hardware connections?
I would appreciate any feedback - Thanks!
Karen Rethoret York University, Toronto 416-736-2100 x33289
I have access to a LARGE number (ie several cases) of microscope cover glasses that I would send to anyone willing to pay the cost of shipping.
These cover glasses are #1 and are 24 mm X 60 mm (not what I call a standard size). They are "Bioloid" Brand from Will Corp. Rochester, N. Y. The packaging says "uniform quality non-corrosive".
Return-Path: {Microscopy-request-at-sparc5.microscopy.com} Received: from relay08.mail.aol.com (relay08.mail.aol.com [172.31.109.8]) by air13.mail.aol.com (v36.0) with SMTP; Thu, 11 Dec 1997 00:34:53 -0500 Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by relay08.mail.aol.com (8.8.5/8.8.5/AOL-4.0.0) with SMTP id QAA17813; Tue, 9 Dec 1997 16:47:56 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id LAA13871 for dist-Microscopy; Tue, 9 Dec 1997 11:26:35 -0600 Received: from mail-oak-2.pilot.net (mail-oak-2.pilot.net [198.232.147.17]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id LAA13868 for {microscopy-at-Sparc5.Microscopy.Com} ; Tue, 9 Dec 1997 11:26:32 -0600 Received: from ns.nissei.com (mail.nissei.com [198.70.112.150]) by mail-oak-2.pilot.net with ESMTP id JAA19369 for {microscopy-at-Sparc5.Microscopy.Com} ; Tue, 9 Dec 1997 09:39:37 -0800 (PST) Received: from ccmailx.nissei.com ([137.168.173.249]) by ns.nissei.com (Netscape Mail Server v1.1) with SMTP id AAB11689 for {microscopy-at-Sparc5.Microscopy.Com} ; Tue, 9 Dec 1997 09:24:03 -0700 Received: from ccMail by ccmailx.nissei.com (IMA Internet Exchange 2.02 Enterprise) id 48D82E50; Tue, 9 Dec 97 11:41:57 -0600
Return-Path: {Microscopy-request-at-sparc5.microscopy.com} Received: from relay08.mail.aol.com (relay08.mail.aol.com [172.31.109.8]) by air13.mail.aol.com (v36.0) with SMTP; Thu, 11 Dec 1997 00:34:53 -0500 Received: from Sparc5.Microscopy.Com (sparc5.microscopy.com [206.69.208.10]) by relay08.mail.aol.com (8.8.5/8.8.5/AOL-4.0.0) with SMTP id QAA17813; Tue, 9 Dec 1997 16:47:56 -0500 (EST) Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id LAA13871 for dist-Microscopy; Tue, 9 Dec 1997 11:26:35 -0600 Received: from mail-oak-2.pilot.net (mail-oak-2.pilot.net [198.232.147.17]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id LAA13868 for {microscopy-at-Sparc5.Microscopy.Com} ; Tue, 9 Dec 1997 11:26:32 -0600 Received: from ns.nissei.com (mail.nissei.com [198.70.112.150]) by mail-oak-2.pilot.net with ESMTP id JAA19369 for {microscopy-at-Sparc5.Microscopy.Com} ; Tue, 9 Dec 1997 09:39:37 -0800 (PST) Received: from ccmailx.nissei.com ([137.168.173.249]) by ns.nissei.com (Netscape Mail Server v1.1) with SMTP id AAB11689 for {microscopy-at-Sparc5.Microscopy.Com} ; Tue, 9 Dec 1997 09:24:03 -0700 Received: from ccMail by ccmailx.nissei.com (IMA Internet Exchange 2.02 Enterprise) id 48D82E50; Tue, 9 Dec 97 11:41:57 -0600
There is an excellent paper that I would be pleased to send you a copy of=
which deals with ion milling of hard coatings. The paper is:
"Transmission Electron Microscopy Characterization of Hard Coatings and Films; Sample Preparation Aspects and Results" G. Radnoczi, Arpad Barna Research Institute of Technical Physics, Hungarian Academy of Sciences. =
Surface and Coatings Technology, vol 80 (1996) pp 89-95.
This paper deals with several materials such as: =
10u diamond film on silicon mechanically alloyed Al-Cu Powder SiC fibers in plastic SiC/Si TiN/Si
All of the work was done with an IV3 Research Grade Ion Milling System which is offered by South Bay Technology. Therefore, I do have a financi= al interest in this posting. Nonetheless, it is a very good paper. I also have several other papers by Dr. Barna et al which deal specifically with=
ion milling difficult materials. I think they would be valuable refernce=
} } } } } 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 Adam Papworth } ------------------------------------------------------------------------=
The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
Please could somebody help me.
I am trying to make a TEM specimen of Diamond, I have a Gatan PIPS at my disposal, but to use it my Diamond wafer has to be less than 100 microns thick. At the it is 1mm thick. Any suggestions on how to thin the Diamond wafer? The Diamond is actually poly-crystaline CVD
Thank you in advance Adam Dr Adam Papworth Dept Materials science & Engineering Ashton Building The University of Liverpool L69 3BX
Phone No 0151 794 5372 Fax No 0151 794 4675 E-Mail adamp-at-liv.ac.uk {
SOBOCIG wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } In short, I would like to inquire as to the methods and chemicals that people } are using for cleaning their filament assemblys on their electron microscopes. } Especially the final step. } } When changing our filament for our TEMs, we have in the past cleaned any } tarnish or deposits from the filament assembly pieces using a mild polish, then } sonicated the parts in a detergergent solution, followed by water rinses, } sonication in a few rinses of ETOH, then finally sonication in a freon-based } ultra-precision cleaning solution. (The used freon solution is dumped into its } own waste bottle.) } } We are considering changing this process (especially in consideration of the } availability and disposal of the freon solution) and are wondering what other } labs use for cleaning their filament assemblys thoroughly. We are especially } interested in the final cleaning step (removing any leftover residues). } } Thank you for your input. Don't be afraid to be brief. } } Gregg Sobocinski } Parke-Davis Pharmaceutical Research } Ann Arbor, Michigan } USA } Sobocig-at-aa.wl.com
Gregg I use this procedure on SEM gun parts, apertures, aperture strips and any other critical parts: 1.) Clean with 1u diamond paste. Chuck Garber tells me that his has no silicones, unlike the metalographic pastes. I don't know for certain, but it works fine and he has the best price. I do third party service and an 18 gram syringe lasts me for years.
2.) Clean (ultrasonic) with Joy dishwashiing liquid and hot water. I understand that the Proctor & Gamble labs clean their critical AAU parts in this and find no detectable residue (-at- ppm or better levels).
3.) Rinse under hot tap water to remove all detergent residue (cold tap water has also always worked for me when that is all that is available).)
4.) If available, rinse with distilled water or DI water. If not avaliable, don't worry because I"ve never had any trouble finishing with tap water.
5.) Blow dry to avoid drying residue, especially in critical areas (wehnelt opening, anode opening, etc.). This IS important!
I'm going to get spammed on the use of water, but the reasoning is as follows: All organic solvents come in contact with plastics of various types, most of which contain plasticizers. Plasticizers will and definitely do contaminate e-beam columns. They're a lot like Apiezon grease. Keep them out of your vacuum system. Water will not polymerize, will not contaminate and will not stay in your vacuum system. It may take some time to pump it out, but when it's gone, it's gone and there is no trace of it left on your apertures, baffles and other critical parts. It is non-toxic (at least so far), it is non-flammable, it is readily available and it is cheap. The only negative effect that I've seen is the temporary increase in total pressure in the vacuum system. Before you anti-water people get wound up, have you ever put a resudual gas analyzer on your microscope? Try it and I guarantee, even with a turbo pump, you'll have at least 90% water. If you're using a diffusion pump you will have 97% or better water in your vacuum system (assuming that it's not leaking). That's why I don't worry about water. For those who are using cryopumps, I'll cry "uncle" (but how do you put up with the vibrations?).
Ken Converse owner Quality Images 3rd party SEM service
Dennis Collins wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Subject: Time:5:39 PM } OFFICE MEMO RE} Cooling water problems Date:12/10/97 } } Hans Brinkies wrote, } "I am still using an old ETEC Autoscan (SEM, vintage 1973). It is } still working well after more than 12 000 hrs of usage and usually we } get the results that we want. } } "However, a calcium containing deposit has been forming } in the cooling water supply (in Cu tubes, in cooling coils around } diff.pump, in heat sinks, ect). The microscope was donated to us } several years ago but was not connected for the last 18 months } to the recirculating water system in our laboratory ( we are using } filtered tap water). The water flow has now been reduced drastically } over the last few weeks and I fear that the 'pipes' may eventually } totally block up. } } "What is the best (and safe) way to reduce or remove this deposit. } Back-flashing was only partially successful. } } "Any suggestion ? } } "Thank You } } "Hans Brinkies } SWINBURNE, University of Technology } School of Engineering and Science } Electron Microscopy Laboratory } HAWTHORN, 3122, Australia" } **************************************** } Hans, } The same problem occurs in lots of water-cooled, high power equipment such } as vacuum tube amplifiers, x-ray tubes, and particle accelerators. } For components with copper cooling water passages we use only low } conductivity water (min of 750K-ohm-cm: 1 Megohm-cm is better) in a } closed-loop cooling system. Even then deposits will accumulate. } We use water flow switches to ensure at least a minimum water flow in } sensitive equipment, and try to keep an eye on the pressure drop across each } water cooling circuit, back-flushing with a dilute solution of Sulfamic acid } when we see a clear rise in the pressure drop at the required flow. } Hope this helps keep your ETEC Autoscan running. } Dennis Collins } DGCollins-at-lbl.gov
Hans
Some how I didn't get the original message and don't have your e-mail address and so can't reply directly, but here goes:
How much flow can you get? The system only needs 4 to 6 gallons/hour but you should be able to get more than 10 gph at 20 psi and a good deal more at higher pressures. You may be able to do an acid flush on the whole thing but I would only flush the whole thing if the problem isn't localized. The ETEC is fairly easy to trouble-shoot in the plumbing area and you should find where your blockages are, first. They can be most anywhere, but start with the DP, the nylon "J" tube following and the 1/4" copper tubing from there to "water out". Also, sometimes it is only the fittings, not the tubing, so just replace them. Most likely the DP is plugging up. Remove the DP from the system. Remove the 1/4" PolyFlo x 1/8 mpt elbow from the outlet end and scoop out as much muck as you can. Try water or air (at 60 - 100 psi) from the top. If the flow remains restricted, pass some HCl (10%-30%) through under low pressure, about a 1 to 2 foot head, and be careful of the splatters. Follow with water and repeat if needed. If the nylon "J" tube is the problem, you can try and clean it or just replace it. Polyethylene tubing is not recommended here due to the high temperatures, but can be used in a pinch. Either type of tubing can be bent in boiling water, then cooled to form a permanent "J". If the copper tubing between the nylon tubing and the outlet is plugged, replace it with 1/4" refrigeration tubing. Once you get your system cleaned out, periodically flush it. The easiest way is to turn off the water, disconnect the "water in", and attach the "air" to the "water in" to blow out the lines. Reconnect the lines correctly and run the water at as high a pressure as you can. Repeat this until the lines run clear. AFter you get familiar with this, you won't even bother to turn the DP off because it can be done so quickly.
Ken Converse owner Quality Images 3rd party SEM service (ETECs are our specialty)
Just an update from an electron microscopist who as a service engineer has probably cleaned more electron guns round the world than very many others.
As many will know I have been involved with the maintenance of electron microscopes for 33 years. Firstly as a service engineer and then through my own training organisation where we train both operators and service engineers. We run regular maintenance courses around the world when we get a good idea of which cleaning materials and solvents are available.
Once again I have been watching the discussions with interest to see if there were many holes in the cleaning explanations. That said may I toss in my few pennies (cents) worth?
TUNGSTEN Gun Systems
The cathode assembly should be cleaned every filament change, the anode every other change and the electron gun at least once a year.
Materials - Almost any metal polish of less than 1 micron may be used to clean electron gun components however it must not be LONG LIFE. Long life additives coat the cleaned item with a polymer that causes chaos in the electron gun. Look out for any indication on the bottle or tube that the manufacturer is claiming that you will not need to clean the metalwork so often after using their product!
Method - Almost more important than the cleaning efficiency is our ability to completely remove the polishing media. So many service call outs are due to problems caused through inefficient removal of the media. For this reason it makes sense to use a metal polish that is easily removed by a solvent for tungsten. In this way we not only remove the metal polish but also clean the areas that are difficult to approach with the polish, nooks and crannies! Also very important is the need to clean without damaging the component, scratching it or placing cotton hairs within the "traps" that the manufacturers seem to put in our way. The best cleaning technique is a wet clean, that is to use solutions and an ultrasonic cleaner. In this way the damage that mechanical forces apply to the components are minimised. Sure the cathode aperture may need a little more encouragement to give up its deposit but only do this if the wet cleaning procedure falls short. We like "Silvo" or "Bluebell" or "Brasso", liquid metal polishes that will mix with a dilute ammonia solution to form a cleaning media, but a solution that may be removed with further washes in dilute ammonia. The mix - 10% metal polish in 90% ammonia solution - where the solution is 10% ammonium hydroxide in water. Place the components, one at a time, in the solution with their least important face down wards. Never put gun components together in the solution as they will damage each other. Do not put an aluminium cathode in ammonia as it will go black, oxide! After 20 minutes in an ultrasonic the component should be clean, wash off in running water and run for another 5 minutes in straight 10% ammonium hydroxide in water. Swill off with running water and then wash in alcohol and dry. NEVER throw away your solutions until you have reassembled the cathode as it is quite possible for the small screws to have fallen out and to reside in the debris at the base of one of the cleaning containers. If you do have a deposit remaining in the aperture area of the cathode a little mechanical effort with the cleaning media may be required,
The gun chamber IS important and this should be cleaned through disassembly once a year, particularly with a TEM. Dirty guns hold gas and induce micro discharge which spoils images. Clean the gun chamber with metal polish, remove the metal polish with dilute ammonia and buff up the walls with a clean chamois or dear skin leather. To retain the cleanlyness of the chamber, each time you change a filament buff up the walls with the leather. If the chamber smells, oily-ozone smell, but is not visibly stained, this is the result of discharge and all traces of the smell should be removed with dilute ammonia.
Look after your gun, it is probably the dirtiest area of the microscope, other than the specimen area in a SEM or the camera chamber in a TEM, its state will determine the ultimate performance of the instrument (high voltage stability) and your filament life.
LANTHANOM HEXABORIDE
Technique developed by ANU Electron Microscopy Unit Canberra
Clean the cathode with 25% hydrochloric acid in water by immersing for 60 seconds and then cleaning with a weak alkaline (ammonia or sodium hydroxide). Wash with water and then alcohol before drying.
LaB6 sources should last a long time (1000 hours plus) but they do need an intermediate cleaning session about every 250 to 350 hours. Some people amaze us by getting away with 1100 hours without cleaning but this is the exception not the rule.
Good luck!
Steve Chapman, Senior Consultant, Protrain, Oxford ,UK Tel & Fax +44 (0) 1844 353 161 web page - http://ourworld.compuserve.com/homepages/protrain
I about to purchase a double tilt heating stage (~1000=83C) and would like any suggestions or critics of Oxford vs Gatan (or any others). This would be for use in a JEOL 2010 with +_ 30=83 of tilt in both the x and y. In particular I would like info regarding temperature stability and accuracy, TEM sample stability, extra EDS signals, "smoothness" of tilting, durability of the holder, sample thickness limitations of the holder, etc. I know I could get info from the vendors but they sem to think they are perfect and the others are substandard. Thanks in advance
Brad Storey Materials Scientist Argonne National Lab - West P.O. Box 2528 Idaho Falls, ID 83403 Ph. 208-533-7685 (office) Ph. 208-533-7439 (lab) =46ax 208-533-7683 =05brad.storey-at-anl.gov
We're looking for a "highest-resolution at low-cost" laser printer for SEM micrographs. LaserMaster has an 1800 dpi printer at about 5-10 cents per page. Experience/suggestions appreciated.
JB
Jacob Bastacky, MD Room 116 Donner Lawrence Berkeley Laboratory EMail: sjbastacky-at-lbl.gov University of California Telephone: 510.486.4606 Berkeley, California 94720 FAX: 510.486.4750
Dear sir, I'm read your home page and known your address on it.My name is Ms.Vimonwan from Faculty of Public Health,Burapha University.I'm Anatomist and lecturer.I'm beginning study about skin Thai frog and I hope to know about processed some skin frog for TEM,SEM? Because skin frog have thinkness of mucous layer.I don't know" How to wash -out ?" What is the best fixative to keep it? How long! If you don't mine,please introduce me about that and name of text book to improove my work.Area of skin on thumb in male frog and behind axilla in female frog. I will to comparative in this area in during breeding season and non-breeding. Thank you very much for your kindness. Finally,I'm looking forward to hearing from your soon. Yours sincerely, Ms.Vimonwan
My address: Ms.Vimonwan Nakiem Faculty of Public Health, Burapha University, Chonburi 20131 THAILAND
Hello All I would appreciate suggestions to following matter: in } 5 years old TEM microscope (particularly CM-20 Philips, with Riber IGP) the ion getter pump is getting old and the pumping times increase(specially after admitting air) - for many reasons I am aware of, but one of them is erosion of Ti electrodes inside IGP. 1) When would You decide to exchange the pump body - what level of decrease in performance ? 2) Did You exchanged allready by Yr machine the IGP pump ? - if yes - after what period of operation, and what were the symptoms ? Thanks in advance for any info from experience and practice.
regards Krzysztof M. Herman LabSoft S.c. 05-500 Piaseczno, ul. Kosciuszki 21, Polska tel/fx: (48 22) 7502024, 7502028, 757067, mobile: (48 90) 213438, (48 90)299748 fax: (48 22) 483787, Email: labsoft-at-labsoft.com.pl zapraszamy do http://www.labsoft.com.pl/ ------=_NextPart_000_01BD06D9.1534E5A0 Content-Type: text/html; charset=ISO-8859-2 Content-Transfer-Encoding: base64
Ronnie: Are you indifferent to know who wrote and sponsored a research project? Do you thing that in our society it is unimportant to know who recommends a product? Do you think its o.k. for a person with obvious commercial interests to edit a "Trade journal" without at least advising readers of those commercial interests?
These things matter a good deal, otherwise the big buck will rule all and not just most things. "Childish"? I have not been called that for some decades! I would, however, advise that ad hominem (against person rather then the argument) should not be used in a professional forum. "Had gumption"? Well, our online amounts to 11 megabytes. Over half of the "pages" are services like: MSDS, User Notes and Links. Tell me when you find a better catalogue, I love to learn.
Our catalogue is obviously a commercial site and its not masquerading as a society's site. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au ---------- } Jim, } Don't be so childish. If your company had the gumption and interest to } support the profession you serve, perhaps you would have had the insight } to establish such a site as Steven has. } Ronnie Houston } Dallas, TX
} Date: Friday, 12 December 1997 3:26 } Jim Darley wrote: } } } } Its fair and reasonable for Steven Slap to provide this information, but } } why does he not indicate that he is the editor of the suggested histology site?
} } Sure his name appears on that site and his company is shown as a sponsor (a } } cheap form of advertising), but no connection is made and most visitors } } would not realise that Steven Slap is the manager of the sponsoring firm and } } thus has a commercial interest: In essence it is a commercial site. } } Predictable his company is the only consumable supplier company shown. } } Nothing wrong with his sponsoring or editing such a page but visitors to } } the page and readers of this listserver are entitled to know not just who } } is sponsoring what but what commercial interests the editor has. Its known } } as propriety! } } Jim Darley } } } } ProSciTech Microscopy PLUS } } PO Box 111, Thuringowa QLD 4817 Australia } } Phone +61 77 740 370 Fax: +61 77 892 313 } } Great microscopy catalogue, 500 Links, MSDS, User Notes } } ************************ http://www.proscitech.com.au } } } } ---------- } } } From: Energy Beam Sciences, Inc. {ebs-at-ebsciences.com} } } } Dear fellow microscopists, } } } } } } At 08:55 AM 12/10/97 +0100, Heinz Fehrenbach wrote: } } } } can anyone of this newsgroup give me information about and subscription } } } } procedure to any related newsgroup ? In particular, I would appreciate } } } } to hear whether there is a special newsgroup discussing } } immuno-histology, } } } } -fluorescence, -blotting, and pathology subjects. } } } } } } There is an immunohistochemistry list, called ipox-l } } } } } } To subscribe, send a message to majordomo-at-pathology.stanford.edu and } } write } } } subscribe ipox-l in the body of the message. I believe that there is } } also a } } } pathology list, and instructions for this are available at "The } } Histotech's } } } Home Page" (http://www.histology.to) } } } } } } Best regards, } } } Steven E. Slap } } } ******************************** } } } Energy Beam Sciences, Inc. } } } Adding Brilliance To Your Vision } } } ebs-at-ebsciences.com } } } http://www.ebsciences.com/ } } } ********************************
Krzysztof M. Herman =20 LabSoft S.c. 05-500 Piaseczno, ul. Kosciuszki 21, Polska tel/fx: (48 22) 7502024, 7502028, 757067,=20 mobile: (48 90) 213438, (48 90)299748 fax: (48 22) 483787, Email: labsoft-at-labsoft.com.pl zapraszamy do http://www.labsoft.com.pl/
Dear Krzysztof, I don't know about your special type of Riber IPG; I have in my lab a = LEYBOLD-HERAEUS IPG, which has been changed as a whole (Ti-netting) 2 = times now (within 17 years of use). In addition, 2 times we could turn = the Ti-nettings 180 degrees round (after cleaning). In the manuals = provided by LEYBOLD for the IG-pump there was indicated a life-span of = about 47.000 working hours, provided vacuum better than 1.0x 10 to the = minus 6 mbar, and it was suggested that it was more economic to change = the whole pump after those 47.000 hours of pumping. So I am convinced, = you should have an instruction manual on the RIBER IG-pump too, provided = by the TEM-dealer (Philips, or any else). If you don#t get to a solution, try the following Compnay/+address for = more information on re-building your particular IG-Pump:
DUNIWAY STOCKROOM CORP. 13?5 Space Park Way MOUNTAIN VIEW, CA. 94043 USA Phone: USA/650/969-8811 Fax: USA/650/965-0764 or visit their www-Site:
No commercial interest in products/product lines, company/-ies, if such = names are mentioned or such are refered to. In this case I am only a = catalogue holder.
Best regards and Season's greetings, Merry Christmas and calm holidays Best wishes for a HAPPY, HEALTHY, SUCCESSFUL and PROSPEROUS NEW YEAR TO = ALL of YOU
Wolfgang MUSS Department of Pathology, LKA EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
---------- Von: labsoft[SMTP:labsoft-at-labsoft.com.pl] Gesendet: Freitag, 12. Dezember 1997 08:36 An: MSA Microscopy Betreff: Old Ion Getter Pumps...
At 07:57 AM 12/12/97 +0200, you wrote: } yes. i meant carbon coated holey films. } Sara }
Sara,
Here is a method I have used in the past. It's a hassle, believe me, but it usually works. It also makes you realize why purchasing holey films commercially costs so much---they're not easy to make.
1) If you can find it, purchase a product called "Victawet" (I believe Electron Microscopy Sciences carries it). This comes in very small vials and it lasts forever. It serves as a lubricant to release plastic films from glass slides in the following steps.
2) Get some high quality glass microscope slides. We have found that Esco brand slides work more reliably than any other kind. Don't know why. The ones with one frosted end are useful for keeping orientation.
3)Polish these slides until they gleam and show NO contamination upon close examination. Do this even if they are "precleaned".
4)Take a piece of Victawet about the size of a cooked grain of rice and put it in a tungsten basket in a vacuum evaporator. Arrange as many CLEAN glass slides around it facing the basket. A distance of several to many centimeters is fine----i.e., distance is not too critical. It may be possible to make a custom rack for this purpose out of plastic or metal---the material used should not outgas too much.
5) Pump down the evaporator and when high vacuum has been reached, gently increase current to the basket. At a certain point the victawet will start to melt and you will see a fog developing on the slides. (If you use too much current, the piece of victawet may jump out of the basket and you have to start over, so be patient.) This "fog" is what you want. No need to overdo it, a little bit will work fine.
6) Repeat Step 5 until you have as many slides as you need. Use a new piece of victawet for each batch. Store these slides in a container for later use, since they keep for a few weeks.
7) Get some formvar (some people use butvar) in 1,2 dichloroethane (ethylene dichloride). 0.5% or 0.25% usually works. You can order 1% and dilute it, too. Keep this solution in a dessicator. Water in the solution causes problems, so only open it when necessary and for as brief a time as possible. Pour this in a tube wide enough to hold a slide, to a depth just short of the frosted end of the slide. (You can conserve formvar by using a special tube with a constricted lower end. One product is called "Dip Miser" and I think it's sold by Ted Pella. We had a special dipping tube made by a glassblower from a regular glass cylinder fused with a very wide constricted bottom.)
8) Cover the top of the tube with the formvar with something so that the atmosphere inside becomes saturated with the evaporating dichloroethane.
9) Get a container big enough to hold water to a depth of several inches. It should be 8-10 inches in diameter for comfortable working. Fill it to the top with, preferably, double-distilled water.
10) Now you're ready. Take the victawet coated slides and polish them again. They should feel slippery. Clean them until they gleam. Clip the frosted end of the slide with something to hold it, like a paper clamp attached to a piece of wire, and put it in the formvar solution in the tube. Keep it there for about 5-10 seconds, then pull it up and let it drain INSIDE THE TUBE. Only leave the top of the tube uncovered while putting the slide in and taking it out, in order to keep the atmosphere saturated. While the slide is draining, keep something over the tube opening, allowing only enough opening for the wire. The length of time you let the slide drain determines the final thickness of the formvar film. Start at about 10 seconds. If you desire a thinner film, increase the draining time up to about 15-20 seconds. (This is why the atmosphere must remain saturated with dichloroethane inside the tube. If it is not, the formvar just dries with draining properly.)
11) To make the holes, expose the slide to moisture IMMEDIATELY upon removing the wet slide from the dipping tube. We would breathe on it, but passing it quickly over a bath of steaming water might do the same thing. The microdroplets of water in the steam or in your breath make the holes. To repeat, this must be done IMMEDIATELY before the slide has time to begin drying. This is also a good time for prayer, invocations to the ancestors, luck rabbit's feet, and anything else you might find effective in appeasing the gods of holey films.
12) Lean the coated slide up against something in a dust-free area to dry for at least two minutes.
13) Back to the basin of water---take a clean laboratory tissue paper and drag the surface of the water to rid it of oil and dust. You can also put a drop of collodion in amyl acetate on the water, let it form a film, then pick this up and discard it to clean the surface.
14) Take the slide and cut around the outside perimeter of the film with a clean razor blade or scalpel. Then take the slide and, holding the frosted end, SLOWLY dip it into the water. If all goes well, the victawet was good, and you have been virtuous recently, the film will separate from the slide and float on the surface of the water. (HINT: a friend of mine---thanks, Steve---discovered that heating the water really helps in releasing the film. Don't boil it, just warm it up.)
15) You can now take your clean TEM grids and place them carefully on the floating film. When you have enough, take another CLEAN glass slide (no need for victawet this time) and scoop the film up. This is tricky and hard to describe: the idea is to catch the end of the slide on the end of the floating film so that when the slide is pushed down into the water the film will be pulled down with it and against it. When you finish, the grids should be between the formvar film and the glass slide. Put it somewhere dust free to dry. When dry, you place the slides in a vacuum evaporator and coat them with 100-300 angstroms of carbon. Then, you can GENTLY AND CAREFULLY push on the edge of the grid to pop it loose from the film. At this point, you will hopefully have a carbon-coated holey film on a grid, ready for use.
Alternatively, you can use a "domino rack", a piece of metal with small holes in it and two bent ends that serve as legs (kind of like a little bench with holes a few millimeters larger than TEM grids). These can be purchased commercially, or made yourself if you can find the right kind of metal with holes. When the film is floating on the water, before putting the grids on it, slowly lower the CLEAN domino rack onto the film, which will adhere to it by surface tension. Lift it out and put it in a dust free place to dry. The result is formvar film covering a bunch of little holes. The grids are later set upon these films by placing a drop of double-distilled water on the film, placing the grid on the drop, and letting it dry down. The film with grids can then be carbon-coated, or you can coat the individual grids later.
I warned you. This is a good starting procedure and variations can be done at several points, depending upon your circumstances. Things can go wrong at any stage, affected by humidity especially. It's a frustrating procedure and I would welcome any suggestions for making it easier, but it does work if you're patient and willing to experiment a little. If you do get a batch to work, I suggest making a whole bunch at once while luck is with you. You may not be so fortunate next week.
Hope this helps. Let me know if you have any questions, and I'll try to help more.
Randy
Randy Tindall Electron Microscope Laboratory Box 3EML New Mexico State University Las Cruces, NM 88003
I've had a request from someone for information on the wee mites - I think they're mites! - that lurk around our eyelashes. They especially wanted to see a picture. Someone suggested that Discover Magazine had published such a micrograph. I went through a stack of back issues but missed any such article. Does anyone know of a handy source of this information/photo? Thanks.
Carolyn J. Emerson email: cemerson-at-plato.ucs.mun.ca
Biology Department Memorial University St. John's, NF A1B 3X9 Tel: (709) 737-7515 Fax: (709) 737-3018
Fellow Histoneters, I know that this has been discussed but obviously I need more imput. What is everyone doing about using controls for immunofluorescent studies. My understanding is that CAP dictates that a control must be run on each antigen being used. Has CAP provided the source where we can "purchase" these controls? If a Positive Patient is used as a control, are we legally allowed to do this without informing the Positive patient? Please help! Teresa
Brad- both holders are probably very well made, I have experience with the=20 Gatan double tilt, temp. controlled specimen holder, the temp control unit= =20 was very accurate, the stability is somewhat compromised (compared to=20 standard double tilt holders), using the Be locknut and Hex ring reduces th= e=20 chances for any unwanted signal, the vacuum on the liquid nitrogen vessel= =20 needs "recharged" every year or so, the only problem I found (which has=20 probably been corrected by now) is with the precision of the tilt axis,=20 there was a very coarse tilt mechanism for the secondary tilt axis. Ask yourself what you need from your tools, then look at each company's=20 spec.s, and haggle price! -M. Rock
On Thu, 11 Dec 1997, Brad Storey wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } =20 } I about to purchase a double tilt heating stage (~1000=83C) and would lik= e } any suggestions or critics of Oxford vs Gatan (or any others). This } would be for use in a JEOL 2010 with +_ 30=83 of tilt in both the x and y= . } In particular I would like info regarding temperature stability and } accuracy, TEM sample stability, extra EDS signals, "smoothness" of } tilting, durability of the holder, sample thickness limitations of the } holder, etc. I know I could get info from the vendors but they sem to } think they are perfect and the others are substandard. } Thanks in advance } =20 } Brad Storey } Materials Scientist } Argonne National Lab - West } P.O. Box 2528 } Idaho Falls, ID 83403 } Ph. 208-533-7685 (office) } Ph. 208-533-7439 (lab) } Fax 208-533-7683 } =05brad.storey-at-anl.gov } =20 } =20 } =20
I'm trying to find out if there are going to be any good short courses or workshops on molecular biology techniques such as in situ hybridization, apoptosis detection and so forth in the next few months. I know that some companies run these courses as do societies and educational institutions. Any information would be greatly appreciated!
To all those who wrote to me before with suggestions for curing my wrinkle problems with semi-thin sections, I thank you, but the problem still persists, after trying those suggestions.
But I only get the problem really with nerve longitudinal sections. Could any technologists or scientists, or techno-scientists suggest any other plastic other than Epon/Araldite or Spurr that might yield better results, such as Epon or Araldite itself???
Ray, We recently retired our Delta system. Over the years we had troubles with the monitor and tried finding anything else that would work with their video signals. The scan rates are just unusual enough that other monitors can't quite synchronize on the signal, and we tried a few.
I would suggest taking the image from the Kevex and transporting it to another computer of your choice. Kevex had a package for converting images. I wrote similar programs on my own which I would be happy to share.
One program converts IMS and FXM files already collected. A second program grabs the graphic screen image and saves it to a file. It works best if you have TSX+ available for running multiple programs at once.
Then there is the matter of shipping the files to a PC. The Kermit program does okay and the Kevex can be attached to the ethernet, but that is not so cheap a solution. Feel free to ask for more details.
At 08:43 AM 12/11/97 -0500, you wrote:
} Has anybody successfully hooked up the color display output from a Kevex } } } Delta EDX to a personal computer running Windows; if so how did you do } it? Our system uses an old DEC computer and an Electrohome monitor with } } } individual RGB and separate horizontal and vertical sync connections. The } } } only printer output is to a serial connected to an old OKI dot matrix } printer. } } Thanks in advance. } } Ray Haythornthwaite ---------------------------------------------------- Warren E. Straszheim 23 Town Engineering Iowa State University Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-8216
Both web pages are very nice, I just subscribed to the IHC newsgroup, and am looking forward to improving my IHC techniques. Everybody has some kind of interest, don't get so paranoid, thanks for the info., drop the argument.
-Mike Rock no disclaimer, no affiliation, no stress
On Fri, 12 Dec 1997, Jim Darley wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Ronnie: } Are you indifferent to know who wrote and sponsored a research project? } Do you thing that in our society it is unimportant to know who recommends a } product? } Do you think its o.k. for a person with obvious commercial interests to } edit a } "Trade journal" without at least advising readers of those commercial } interests? } } These things matter a good deal, otherwise the big buck will rule all and } not just most things. } "Childish"? I have not been called that for some decades! I would, however, } advise that } ad hominem (against person rather then the argument) should not be used in } a professional forum. } "Had gumption"? Well, our online amounts to 11 megabytes. Over half of the } "pages" } are services like: MSDS, User Notes and Links. Tell me when you find a } better } catalogue, I love to learn. } } Our catalogue is obviously a commercial site and its not masquerading as a } society's site. } Jim Darley } } ProSciTech Microscopy PLUS } PO Box 111, Thuringowa QLD 4817 Australia } Phone +61 77 740 370 Fax: +61 77 892 313 } Great microscopy catalogue, 500 Links, MSDS, User Notes } ************************ http://www.proscitech.com.au } ---------- } } Jim, } } Don't be so childish. If your company had the gumption and interest to } } support the profession you serve, perhaps you would have had the insight } } to establish such a site as Steven has. } } Ronnie Houston } } Dallas, TX } } } Date: Friday, 12 December 1997 3:26 } } Jim Darley wrote: } } } } } } Its fair and reasonable for Steven Slap to provide this information, } but } } } why does he not indicate that he is the editor of the suggested } histology site? } } } } Sure his name appears on that site and his company is shown as a } sponsor (a } } } cheap form of advertising), but no connection is made and most visitors } } } would not realise that Steven Slap is the manager of the sponsoring } firm and } } } thus has a commercial interest: In essence it is a commercial site. } } } Predictable his company is the only consumable supplier company shown. } } } Nothing wrong with his sponsoring or editing such a page but visitors } to } } } the page and readers of this listserver are entitled to know not just } who } } } is sponsoring what but what commercial interests the editor has. Its } known } } } as propriety! } } } Jim Darley } } } } } } ProSciTech Microscopy PLUS } } } PO Box 111, Thuringowa QLD 4817 Australia } } } Phone +61 77 740 370 Fax: +61 77 892 313 } } } Great microscopy catalogue, 500 Links, MSDS, User Notes } } } ************************ http://www.proscitech.com.au } } } } } } ---------- } } } } From: Energy Beam Sciences, Inc. {ebs-at-ebsciences.com} } } } } Dear fellow microscopists, } } } } } } } } At 08:55 AM 12/10/97 +0100, Heinz Fehrenbach wrote: } } } } } can anyone of this newsgroup give me information about and } subscription } } } } } procedure to any related newsgroup ? In particular, I would } appreciate } } } } } to hear whether there is a special newsgroup discussing } } } immuno-histology, } } } } } -fluorescence, -blotting, and pathology subjects. } } } } } } } } There is an immunohistochemistry list, called ipox-l } } } } } } } } To subscribe, send a message to majordomo-at-pathology.stanford.edu and } } } write } } } } subscribe ipox-l in the body of the message. I believe that there is } } } also a } } } } pathology list, and instructions for this are available at "The } } } Histotech's } } } } Home Page" (http://www.histology.to) } } } } } } } } Best regards, } } } } Steven E. Slap } } } } ******************************** } } } } Energy Beam Sciences, Inc. } } } } Adding Brilliance To Your Vision } } } } ebs-at-ebsciences.com } } } } http://www.ebsciences.com/ } } } } ******************************** } } }
This might be off-topic, but the news of a cure for my particularly intense and troublesome headaches was so exciting for me, that I thought I might as well share this with you folks, just in case there are some of you in the lab there suffering needlessly.
Basically I just take 2 Aspirin, and 2 Extra strength Tylenol AT THE SAME TIME. A doctor advised me that there was no problem in doing this, (ie: no adverse effects) because the 2 drugs work differently. So, whereas taking 4 aspirin would not be advised, you can fill in the extra cracks of pain relief with the tylenol. I double checked this with my pharmacist and she says that this is perfectly acceptable, and I'm not doing anything dangerous here.
Anyway, this combination of pain kill completely - blowing your headache into hyperspace, leaving you feel very comfortable.
I need to calibrate the magnification in of a TEM for magnifications above x100k, a range where the usual gratings standards are not so good. The resolution of the TEM is not great, but sufficient to use lattice images of asbestos fibers as a standard. However, I would appreciate to hear suggestions on other suitable magnification standards that can be used under diffraction contrast above x100k. Thank you for your help
Augusto _________________________________________________________________________ Augusto A. Morrone 107D-MEL, P.O. Box 116400 MAIC Materials Science and Engineering University of Florida Gainesville, FL 32611 (352) 392-1497 or 6985 Fax: (352) 392-0390 amorr-at-mse.ufl.edu
Augusto, You need to get a hold of a MaG-I-Cal sample. This was recently written up in the latest Microscopy Today by John McCaffrey. You can find them at SouthBay Technology. It not only allows you to do a mag calibration, but a rotation calibration and camera constant as well. It provides a mag calibration from the lowest to the highest mag on the TEM. I've just recently calibrated my TEM from 10kX to 500kX. -Scott Walck
Scott D. Walck PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of S.D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
I need to calibrate the magnification in of a TEM for magnifications above x100k, a range where the usual gratings standards are not so good. The resolution of the TEM is not great, but sufficient to use lattice images of asbestos fibers as a standard. However, I would appreciate to hear suggestions on other suitable magnification standards that can be used under diffraction contrast above x100k. Thank you for your help
Augusto _________________________________________________________________________ Augusto A. Morrone 107D-MEL, P.O. Box 116400 MAIC Materials Science and Engineering University of Florida Gainesville, FL 32611 (352) 392-1497 or 6985 Fax: (352) 392-0390 amorr-at-mse.ufl.edu
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Question I need to calibrate the magnification in of a TEM for magnifications = above x100k, a range where the usual gratings standards are not so good. = The resolution of the TEM is not great, but sufficient to use lattice images = of asbestos fibers as a standard. However, I would appreciate to hear suggestions on other suitable magnification standards that can be used = under diffraction contrast above x100k. Augusto A. Morrone 107D-MEL, P.O. Box 116400 MAIC Materials Science and Engineering University of Florida Gainesville, FL 32611 (352) 392-1497 or 6985 Fax: (352) 392-0390 amorr-at-mse.ufl.edu
Response There is now a TEM calibration standard that supposedly is good the = entire range of mags on a TEM. I just got one but haven't had a chance = to try it. We got ours from Ted Pella but other supply houses may have = it
Judy Murphy San Joaquin Delta College Microscopy Technology Center Stockton, CA
some of us in the lab here are thanking you endlessly for this combination of pain kill completely - blowing our headache into hyperspace, leaving us feel very comfortable, but what do you thinking to mixingly paracetamol with acetyl salicylic acid ? Or evenly taking more paracetamol/tylenol: does your pharmacist thinking this might hit you liverishly, or grammatically?
Ray
ps for research purposes only, at which pint (sic) in your letter did you take the combination?
At 14:58 -0600 12/12/97, Garry Burgess wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Ray Hicks ________________________________________________________________________ |University of Cambridge |Tel 01223 330149 | |Department of Medicine |Fax 01223 336846 | |Level 5, Addenbrookes Hospital |e-mail {rh208-at-cus.cam.ac.uk} | |Hills Road Cambridge |Web http://facsmac.med.cam.ac.uk | |CB2 |ftp server ftp://131.111.80.78 | |UK | | |_________________________________|_____________________________________|
Just a note to clarify a recent posting: I learned the technique I posted for making holey films from Steve Schmitt at the Center for Electron Microscopy at Southern Illinois University at Carbondale, which is directed by Dr. John Bozzola. I don't know where it came from originally, but I wasn't the one who developed it. Don't want to create the wrong impression.
Randy Tindall 2017 Princess Jeanne Las Cruces, New Mexico 88001-4157
The standard you need is the MAG*I*CAL TEM Calibration Standard.
The MAG*I*CAL is a TEM calibration standard that performs all of the thr= ee major instrument calibrations for a TEM: image magnification; camera constant for indexing diffraction patterns; and image/diffraction patter= n rotation for relating crystal directions to features in the image. =
MAG*I*CAL consists of an electron transparent cross-sectional TEM sample made from a MBE grown, single-crystal semiconductor wafer. When the calibration structure is viewed in a TEM, it appears as a series of light=
and dark layers where the layer thicknesses are accurately known. The calibrated thickness measurements of these light (silicon) and dark (SiGe=
alloy) layers are based on careful TEM measurements of the {111} lattice=
spacing of silicon which is visible on the calibration sample itself, and=
are supported by x-ray diffraction measurements. The layer spacings are designed so that the sample can be used to calibrate the entire magnification range in a TEM - from 1,000X to 1,000,000X. As the sample = is also a single crystal of silicon, the calibrations requiring electron diffraction information such as the camera constant and image/diffraction=
pattern rotation can also be performed easily and unambiguously. One single calibration sample can therefore be used to provide all three of t= he major TEM instrument calibrations at all magnifications and all camera lengths.
South Bay Technology, Inc. supplies the MAG*I*CAL and so I have a defini= te financial interest in promoting its use. I also have copies of other research papers that have been written by the developer, John Mccaffrey, which will provide you with much greater detail. If you have an interest= , please let me know and I'll forward the information to you.
As a matter of additional interest, we can provide batches of the MAG*I*C= AL which are all made from the same wafer which provides the ultimate in calibration uniformity. This has proven to be an ideal solution to large=
organizations who are looking for a "company standard" calibration technique. Please inquire for more information on this service.
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
Dear Netters: =
I need to calibrate the magnification in of a TEM for magnifications abov= e x100k, a range where the usual gratings standards are not so good. The resolution of the TEM is not great, but sufficient to use lattice images = of asbestos fibers as a standard. However, I would appreciate to hear suggestions on other suitable magnification standards that can be used under diffraction contrast above x100k. Thank you for your help
Augusto _________________________________________________________________________=
Augusto A. Morrone 107D-MEL, P.O. Box 116400 MAIC Materials Science and Engineering=
University of Florida Gainesville, FL 32611 (352) 392-1497 or 6985 Fax: (352) 392-0390 amorr-at-mse.ufl.edu
Jill Craig wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hi all, } } If anyone is looking into this topic or knows of any relevant } references or SEM methods, I would greatly appreciate the } information. I have scoured our meager library and the current } contents database with virtually no luck. My kingdom for } access to the science citation index. } } Thank you so much for any aid you can provide. } } Jill Craig } University of Northern British Columbia Jill,
I would suggest you contact John Delly through the McCrone Institute in Chicago. John has a wealth of info on wood.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108-2838 USA PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com **************************************************** Microscopy/Microscopy Education America's first consortium of microscopy experts offering customized on-site training & applications solutions
} Jill, } } I would suggest you contact John Delly through the McCrone Institute in } Chicago. John has a wealth of info on wood. }
The full company name (for directory assistance, etc.) is McCrone Research Institute. They are at 2820 S. Michigan Avenue, Chicago, IL 60616. 1-312-842-7100 (voice) 1-312-842-1078 (fax). http://www.mcri.org
-- ********************************************************** Stephen A. Shaffer sshaffer-at-microdataware.com MicroDataware http:www.microdataware.com (Under reconstruction and temporarily out of service) Personal stuff: steve_shaffer-at-compuserve.com http://ourworld.compuserve.com/homepages/steve_shaffer/ **********************************************************
Ray, lost your address but my client provides a software conversion package, KV2WIN whick will handle all your conversion problems for both images and spectra. For more info;
Bob Hessler Hessler Technical Services PHONE/FAX: 203/358-0266
Would it be possible to give me the address/phone/email of Lipshaw (Detroit) - I am interested in using a permenant mount for confocal specimens and I can't seem to find a European source for Permafluor.
thanks
Mark Auty
Dairy Products Centre Fermoy Co. Cork Ireland mauty-at-dpc.teagasc.ie
Salzburg, 14th of December 1997, 11.00 p.m. local time
CONCERN: CHEMICAL ELOXATION OF CORRODED ALUMINIUM-SURFACES
Dear all out there, I greatly would appreciate any suggestion or idea for solving the = following problem: in our Histology-Lab we use a "fixator" apparatus (for rapid fixation of = small tissue specimens contained in baskets in a cylindrical container, = made from chrome-nickel-steel, by means of alternating application of = pressure and vacuum in formaldehyde solution). The chrome-nickel-steel cylinder is durably fixed to a "collar" made of = eloxated aluminium (they cannot be separated for what reason ever), = which on its top serves as base for a sealing lid (made from plastic) = with an O-ring. Due to working with the specimen baskets (which turn out to be almost as = wide in diameter as the narrow cylinder's diameter) the eloxation of the = Aluminium-collar has been distorted and corrosion has been initiated.=20
The manufacturing company told us that there is no chance to overcome = that corrosion, the only way for stopping it could/would be to overlay = the corroded areas (after thorough cleaning: with what, if not alcohols = or acetone??) with silicone paste. Corrosion should be stopped then due = to exclusion of air/O2. The container-combination (chrome-nickel-steel - Aluminium) described = and used in the fixator now has been replaced by an other construct = which doesn't fit into the old apparatus. Also, one cannot buy the old = container as a sparepart, because it is not available any more. Since corrosion maybe goes on despite pasting silicone (as a byproduct = of condensation acetic or another acid will be formed) over the corroded = areas I am not sure about risks of a breaking of the Al-collar due to = pressure later on. Last chance would be to buy a new "fixator" which amounts appr. US$ = 3000.-
So my question:=20 is there ANYBODY WHO KNOWS OR HAS EXPERIENCE how to SEAL corroded = ALUMINIUM SURFACES, most elegantly by "artificial, CHEMICALLY INDUCED = ALUMINUM-ELOXATION" (procedure??)
Thanking you very much for your considerations best regards with my and our best wishes for a MERRY CHRISTMAS and A HAPPY, HEALTHY, PROSPEROUS and SUCCESSFUL NEW YEAR To you all and especially to you, and you
Dr. Wolfgang MUSS Department of Pathology, LKA EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
Dear colleagues…. Let me present you with a challenging scenario…. I am trying to Golgi stain some fixed mouse brains that have been sent to my lab…thus far I have tried using a Rapid Golgi and a Golgi Kopsch variant with no success….it turns out that although the brains were supposed to have been fixed in 10% neutral buffered formalin, instead they accidently used 10% formaldehdye (a 10% formalin solution contains only 4% formaldehyde). I am convinced that the excessive concentration of formaldehyde has contributed to the difficulty in obtaining successful Golgi staining. So, my question is --- Is there any way that the brains can still be saved for Golgi impregnation?? Might it help if the brains were rinsed in running water for a lengthy period and then put into the correct formalin solution?? Any thoughts or suggestions would be welcomed…. Thanks in advance… Ron Mervis ~~~~~~~~~~~~~~~~~~ Ronald F. Mervis, Ph.D. Neuro-Cognitive Research Labs RonMervis-at-aol.com
Have you thought of finding out the reason of your headache and fix that rather than stuffing yourself with drugs? And yes, it is really off- topic. ----------
This might be off-topic, but the news of a cure for my particularly intense and troublesome headaches was so exciting for me, that I thought I might as well share this with you folks, just in case there are some of you in the lab there suffering needlessly.
Basically I just take 2 Aspirin, and 2 Extra strength Tylenol AT THE SAME TIME. A doctor advised me that there was no problem in doing this, (ie: no adverse effects) because the 2 drugs work differently. So, whereas taking 4 aspirin would not be advised, you can fill in the extra cracks of pain relief with the tylenol. I double checked this with my pharmacist and she says that this is perfectly acceptable, and I'm not doing anything dangerous here.
Anyway, this combination of pain kill completely - blowing your headache into hyperspace, leaving you feel very comfortable.
Thanks for all who responded to my query. Especially those who were brief, but I learned a little bit from all.
For those of you interested in results of the survey, here's my abbreviated, paraphrased summary. I have focussed on the solvents involved, since that was the focus of my question. (My apologies if I misrepresent any procedures due to my brevity.) --- --- --- --- --- --- I received lots of information on polishes, and it seems that matching the solvents to the type of polish that needs to be removed is a good approach. (Some polishes clean up very easily with 10% ammonia solutions.)
It was suggested to use a 10% polish (Silvo, Bluebell, or Brasso) with 90% ammonia solution. This will turn aluminum pieces black, though.
Potassium Hydroxide (KOH), followed by dH2O and ethanol is a popular way to remove oxidation residues without the need for polish. (Cautions with this technique are that brass may discolor a bit, but it doesn't seem to cause any ill effects to the operation of the gun.)
It seems that drying the assemblys with heat lamps or hair dryers after rising in acetone, isopropanol or ethanol prevents the residue that sometimes forms. It appears that this residue may be from water condensing out of the air onto the assembly, which is cooled from the solvent evaporating.
In at least one case, someone uses dH2O as the final 'solvent' and dries with a hair dryer. The reasoning is that water vapor is the kindest of impurities to have in a column, compared to plastics and other chemicals that may dissolve in other solvents and may become deposited on the assembly during drying.
10% oxalic acid was also recommended as an added cleaning step.
--- --- --- Any more details about suggested procedures can be forwarded to you if you contact me before the end of January. After that, the information will be deleted.
Gregg Sobocinski Parke-Davis Pharmaceutical Research Ann Arbor, Michigan USA Sobocig-at-aa.wl.com
IN SITU HYBRIDIZATION RT-PCR WORKSHOP INTERDISCIPLINARY CENTER FOR BIOTECHNOLOGY RESEARCH UNIVERSITY OF FLORIDA FEBRUARY 5 & 6, 1998
THE CONTENT: In situ Hybridization (ISH) is a powerful method for the localization and quantitation of specific messenger RNA in single cells and within the natural tissue geometry. It has become the method of choice in the histopathological study of gene expression. When the technique is combined with PCR, mRNA can be detected at very low levels. ISH is also used for cellular detection of viral DNA and as a tool to obtain cytological information on the location and alteration of genomic sequences in chromosomes (FISH, PRINS).
This two-day workshop provides participants with hands on experience on the methodology and principles for the in situ RT-PCR detection of low abundance target sequences. The potential applications of the technique will be illustrated by the detection of adrenomedullin mRNA from rat brain tissue sections. In an additional experiment, participants will utilize ISH hybridization to detect viral DNA in infected cells.
Workshop includes lectures on a variety of ancillary techniques. Some of the topics include: Nucleic acid probe technology (antisense RNA probes), non-isotopic labeling and detection, tissue preparation and fixation for in situ hybridization, optimization of in situ methods (silane-coated slides, formalin fixation, protease digestion, DNase digestion and 4.5 mM MgCI2), fluorescent in situ hybridization (FISH, PRINS) mapping, cellular detection of viral DNA, etc.
THE LOCATION: Communicore Health Science Center, University of Florida, Gainesville, FL
FOR INFORMATION & REGISTATION: education-at-biotech.ufl.edu phone: (352) 392-8408 fax: (352) 392-8598
FEE: non-student: $200.00 FL Graduate students: $100.00 Registration Deadline: January 15, 1998 $25 Late Registration fee applies after deadline.
I hate to burst your bubble, but you have just re-discovered Exedrin! :-)
At 02:58 PM 12/12/97 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Does anyone have experience with TEM JEOL 1010 in dark field mode with SHP 10 OL pole piece. Im not satisfied by the results obtained with a SAP 10B OL pole piece of the 1010 JEOL. What is your point of view ? Does SHP pole piece could change something in quality of these images in dark field mode ?
Thanks for help.
P-Yves
E-Mail:sizaret-at-med.univ-tours.fr
Pierre-Yves Sizaret laboratoire de Microscopie Electronique UFR Medecine 2 bis Bd Tonnelle 37032 Tours cedex FRANCE
Dear Scott, With reference to the enquiry from Richard Fonda on the thinning of GaAs, here are some points from our experience. Br in methanol is good for the preparation of plan view specimens of GaAs, but I consider 5% is too strong a solution and likely to cause roughening of the surface. 2% is adequate and slow thinning can be achieved at a concentration of 0.5%. Be careful when adding Br to methanol as concentrated solution can explode.It is also best to use a fresh bottle of methanol as it should be dry. An alternative, which we actually use, is Cl in methanol. This can be used by bubbling the Cl into the methanol. Great care must be taken as at too strong a concentration it will set itself alight. We judge the concentration by the colour which goes from pale yellow to a stronger yellow/green as it increases. Both Br and Cl are hazardous materials and should be used in an adequate fume cupboard.
Pete Augustus Tel +1327 356362 Caswell Analytical Services Fax +1327 356775 GMMT Caswell Towcester Northants NN12 8EQ UK
} The absolute best, fastest, and cheapest way to prepare non-site specific } cross section samples with this type and thickness of metallization on GaAs } is with the small angle cleavage technique. There is a detailed pictorial } outline of the technique in the MRS Specimen Prep for TEM of Materials IV, } Vol 480 that just came out. The authors are myself and John McCaffrey who } developed the technique and has several pubs out on it. The technique } requires very little in terms of equipment that is not usually found in a } lab. Southbay Technology is selling a starter kit that has all the required } supplies. I can typically prepare about 9 samples in about an hour and } exchange and examine a sample within about 5 minutes to see if it is good. } A big advantage to this method is that there is no need to worry about } contamination of the sample. I've used this on GaAs and other materials and } examined the samples in field emission microscopes with no problems. I do } use electronic grade acetone and double rinse them. } } For the plan view samples, you will have to dimple and ion mill because of } the metallization. Don't forget to protect the good side from ion sputtered } material. } } } If you didn't have the coating and just wanted to make plan view samples } from the GaAs, I'd recommend chemically polishing the GaAs from the backside } after mechanically thinning to about 100 um to make the plan view samples. } Peter Goodhew showed me an inexpensive way to do this. I think that the } solution used was 5% bromine in ethyl alcohol (it might have been methanol) } which was dripped onto the sample from a burette. The sample was low } temperature waxed to a coverslip slide that was put onto a Teflon pedestal } with a small amount of vacuum grease mounted in a plastic cup. The grease } was not exposed to the solution because it was in the middle of the } coverslip and far from the edges. The cup was tilted at an angle of about } 30 degrees and was rotating with the use of a small DC motor. A } stereomicroscope was used to help determine when the sample was perforated. } } } You could use this method instead of dimpling if you stopped the process } before perforation and then continued the thinning process with ion milling. } The disadvantage is that you will not know the thickness left to ion mill. } } I hope this helps. } } -Scott Walck } } Scott D. Walck, Ph.D. } PPG Industries, Inc. } Guys Run Rd. (packages) } P.O. Box 11472 (letters) } Pittsburgh, PA 15238-0472 } } (412) 820-8651 (office) } (412) 820-8161 (fax) } } } "The opinions expressed are those of S.D. Walck and not of PPG Industries, } Inc. nor of any PPG-associated companies." } } } ---------- } From: Richard Fonda } To: Microscopy } Subject: TEM GaAs prep } Date: Thursday, December 11, 1997 9:46AM } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
INTERNATIONAL COURSES OF LIGHT MICROSCOPY, PHOTOMICROGRAPHY AND LASER SCANNING CONFOCAL MICROSCOPY GARGNANO (Lake of Garda) October 1998
The Course is a post-graduated theoretical/practical course, with propedeutical lectures and practical stages on microscopy, photomicrography and confocal microscopy. The course will take place in Gargnano (Lake of Garda) in October 1998.
Further information and registration details will be found at the following Web address.
http://imiucca.csi.unimi.it/endomi/micro.html
Thank you Paolo Castano
_____________________________________________________ Prof. Paolo Castano UNIVERSITY OF MILAN INSTITUTE OF HUMAN ANATOMY - CHAIR OF HUMAN ANATOMY FOR PHARMACY Via Mangiagalli, 31 - 20133 Milan (Italy)
Anybody has an idea how to stain starch in Chlamydomonas?
I tried (suggested in the list recently) without succes: Nile Blue, Safranin, ConA-FITC all for fluorescence. (Nile Blue was in water, sometimes after bleachingwith Na-hypochloride. Safranin coming from EtOH or also in water. All after fixation with glutaraldehyde, sometimes also after Triton. ConA stained many things but not starch).
Something else to try? Polarisation Microscopy doesn't work, J2KJ also not (I don't know really why ...)
Arthur Dr. Arthur Schuessler University of Heidelberg Zellenlehre Im Neuenheimer Feld 230 D-69120 Heidelberg Germany
The starch staining for LM on algae samples is generally done by KI+I2. Under LM the starch particals appear as dark-blue color and nothing alse can appear as this color. Acturally the usefull reagent is I, KI is used to increase the soluability of I2.
****************************************** Zhiyu Wang Electron Microscope Lab and Imaging Center Western Kentucky University(WKU) Bowling Green KY 42101
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Anybody has an idea how to stain starch in Chlamydomonas? } } I tried (suggested in the list recently) without succes: Nile Blue, } Safranin, ConA-FITC all for fluorescence. } (Nile Blue was in water, sometimes after bleachingwith Na-hypochloride. } Safranin coming from EtOH or also in water. All after fixation with } glutaraldehyde, sometimes also after Triton. ConA stained many things but } not starch). } } Something else to try? Polarisation Microscopy doesn't work, J2KJ also not } (I don't know really why ...) } } Arthur } Dr. Arthur Schuessler } University of Heidelberg } Zellenlehre } Im Neuenheimer Feld 230 } D-69120 Heidelberg } Germany }
Anytime I even got a whiff of Amyl Acetate (even when working in a fume hood) I would instantly get a very blinding headache. I used to use it as a solvent for Colloidon, a kind of support film plastic. But fortunately I have a better fume hood now, and I also rarely use this now. But I can also get headaches from fumes of epon/araldite and xylol. Even though we keep our polymerization oven in a fumehood, when we open the door of the oven to remove the plastic, some of the fumes escape. Of course safety conditions have improved a lot since days gone by, but sometimes I wonder what residual effects exposure to these chemicals might have on a person.
Garry } ---------- } From: Peggy Brannigan[SMTP:brannign-at-asrr.arsusda.gov] } Sent: 15 December, 1997 12:12 } To: Garry Burgess } Subject: Re: Blinding Headache } } Gerry, } } The subject of headaches may not necessarily be as off topic as one would } think. I had headaches almost daily for years until two things happened: } 1. I stopped using acrolein in my fixative and 2. I moved to a new } building where a.) my office was no longer in the lab itself and b). the } lab met OSHA requirements for # of air exchanges per hour. } } The headaches did not go away entirely but there was a dramatic change-I've } always wondered how many other microscopists suffer headaches. Since we } use our eyes so intensely staring at flickering screens and monitors and } expose ourselves to nasty chemicals it wouldn't surprise me if headaches } are, to some extent, an occupational concern. } } With respect to your tylenol-aspirin discovery , your're right on target } there. I go to a neurologist who basically prescribes the same thing, as } well as a few other medications. Headaches are complex, involving several } different pathways and success often requires taking several different } medications to target different components of the headache. } } Obviously it is best if you can find an environmental or dietary source for } the problem but that's not as easy as it sounds, simply because headaches } are so complex. There are numerous medications available that will prevent } headaches but you have to take them on a daily basis and should be under a } neurologist's care. } } One word of warning...if you take tylenol every day you may experience a } rebound effect where you get a headache when the tylenol leaves your } system. I don't think aspirin does this, but I''m not sure. } } Other things that have helped me: acupuncture, aerobic exercise, muscle } relaxation through biofeedback, and the herb feverfew. } } Anyway, good luck. } } Peggy } } } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have made a list of what I consider to be the more common problems involved in Diagnostic Electron Microscopy as relates to specimen collection, preparation of plastic, sectioning, staining, and photography, for the benefit of outsiders to our lab, so that they might avoid some of these pitfalls. I would be interested in getting feedback from other people who might have other problems in mind that I have not considered.
My guess is you can indeed salvage the brains and still use them. Have they already been processed and embedded in paraffin? If so, you'll have to deparaffinize and rehydrate them, then rinse in running water or soak overnight in a large volume of (preferably) buffer.
Hint: Depends on what kind of tissue processor you have, but it is sometimes possible to run the processor in reverse order to deparaffinize and rehydrate...or, it is also possible to run the specimens through the "purge" or "retort clean" cycle, as well. This does a great job of deparaffinizing the specimens and leaving them in 100% alcohol; from there you can step down to water or buffer.
If you are convinced the extra formalin is the problem, this should do the trick. Unlike glutaraldehyde fixation (which is largely non-reversible), formalin fixation is to a large extent "reversible," in that large amounts can be coaxed out of the tissue by prolonged washing.
Good luck! Let us know how things work out.
Best regards,
Bob ********************************* Robert (Bob) Chiovetti E. Licht Company / 1-800-865-4248 rchiovetti-at-aol.com
********************************* Cryostats / Microtomes / Tissue Processors / Embedding Centers / Slide Stainers / Glass Coverslippers / Microscopes (Representing Leica since 1967) / Fiber Optic Systems / Linear Measuring / Micromanipulation (Linear Encoded, Video) / Image Analysis, Archiving, Capture / Video / Video Printers (Cooled CCD, Digital, RGB, Super VHS, 3-chip) / Vibration Isolation Systems / Programmable Stages / Heating & Cooling Stages
Rick Felten 12/15/97 12:52 PM Does anyone know a way to print several image files in an automatic fashion that would include the file name on the image. I am using windows 95.
Bob Schoonhoven recommended the Fontana-Masson stain for melanin pigment. We have a very fast microwave version of the Fontana-Masson stain. I would be happy to fax it to any interested parties.
Best regards, Steven E. Slap, Vice-President ******************************** Energy Beam Sciences, Inc. Adding Brilliance To Your Vision ebs-at-ebsciences.com http://www.ebsciences.com/ ********************************
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
RE} Safety using microscopes 12/16/97 11:48 AM Dear Keith,
While at ARCO Alaska we too were tasked to find a more ergonomic workstat= ion for palynologists and foram workers alike. We basically looked at the= level of their microscopes, computer screens, keyboards, chairs and heig= ht of desk(s).
Only a couple of people were experiencing problems at the time and the re= st did not care about the workstations because they had no problems. This= means that some people tolerate less than optimum ergonomics than others= .
SOLUTION: We changed the chairs to multifunctional settings type and sugg= ested differents heights of keyboards and monitors, microscopes. Most of = the problems went away except for the one person who had an existing aggr= avated lower back problem. His problem was not fixed by any of these solu= tions, nor could it be.
I personally did not have any problems, but when I changed my chair I not= iced a huge difference in comfort.
Finally, most light microscope users are "slumpers". Notice that they slu= mp down with poor posture even when not at the microscope. This presents = a problem when trying to fix their posture at the microscope and while th= ey are just sitting at their desks.
My personal opinion is that the chair is the single most important adjust= ment to make for workers at the desk and/or microscope.
Best of Luck
John Shane McCrone Research Institute
--------------------------------------
Dear Microscopy Listers
I have been tasked with coming up with something about using light micros= copes without harming yourself! Not so funny, according to two of our now-retired staff who counted plankton etc. for 20+ years. They both suff= ered neck/shoulder/back pain which eased after retirement.
In the UK we have a specific, detailed set of Regulations concerning comp= uter use, another unspecific set covers all workstations e.g. supermarket=
checkouts. A lot of the computer Regs. might be transferred to microscope= set-ups.
I'd appreciate any info, anything to avoid re-inventing the wheel!
} I have been tasked with coming up with something about using light micr= oscopes without harming yourself! Not so funny, according to two of our } now-retired staff who counted plankton etc. for 20+ years. They both su= ffered neck/shoulder/back pain which eased after retirement.
Keith, I have a handout that I use in my teaching to guide students in th= e proper positioning of the microscope and their bodies as they use the instrument. This is particularly important for those who may use the ins= trument for several hours at a time. Perhaps you will find the text of t= he hand-out helpful, though it may not reproduce particulary well in an emai= l message. Here goes:
Good Posture is Essential to Good Microscopy
Let=92s face it. Looking through a microscope is not what our bodies wer= e built for. And, looking through a microscope for an extended period of= time requires an unnatural rigidity of the body. You=92re not moving around, = even a bit, as you normally would. The result can be cramped muscles, especially in the neck and shoulders, and a whole body fatigue that can m= ake 10:00 am feel like 5:00 pm. The solution is correct posture and a pr= oper arrangement of microscope, chair, and body.
=B7 If you=92re going to be doing microscopy for any length of time, insi= st of a good, adjustable, ergonomically-designed chair!
=B7 Adjust the height of the chair so that your feet can rest comfortably= on the floor with an even pressure along the back of the thighs. It may= be advantageous to tilt the seat of the chair slightly down in front to elim= inate any pinch at the back of the knees.
=B7 Note: Adjust the chair height without regard to the microscope height= !
=B7 Adjust the position of the microscope so that you can comfortably gaz= e into the eyepieces without leaning significantly forward. This require= s two adjustments:
=A8 Set the lateral position of the microscope so that it is clos= e to the front edge of the bench with the eyepieces no further away from = you than the front edge of the bench.
=A8 Set the vertical position of the microscope so that it is a b= it high for comfortable viewing. This will normally require elevating th= e microscope on some type of stand on top of the bench. The idea is to for= ce yourself to straighten your back as you draw up to the microscope so t= hat with extended viewing your back is straight and your head in an upright p= osition. Look down into the eyepieces by letting your eyes view at a downward angle. Do not bend the neck to look =93straight ahead=94 into t= he microscope.
=B7 Obtain some type of arm rest for the arm used to focus the microscope= so that you don=92t need to be constantly raising the arm off the bench = surface and so that you are not tempted to say, =93Oh, the focus is good enough,=94= to yourself while you work. Constant, continuous focusing of the micros= cope is essential to minimize eye strain. (As an aside, proper setting of int= er-pupilary distance and eyepiece parfocalization are also essential to minimizing eye strain.)
=B7 After you=92ve established the optimal height for your microscope by = propping it up on phone books, this mornings paper, or whatever, you may = wish to have a permanent stand fabricated to the right height. Consider incorpor= ating one or two arm rests into the design. Go on, it won=92t cost you m= uch and it will make a world of difference in your comfort as you work. No p= oint growing old and crooked before your time!
=B7 Take a break from time to time. Get up, walk around, stretch arms, b= ack, neck, and legs to remove any =93kinks=94 that may be forming before = they become a bother.
-- ********************************************************** Stephen A. Shaffer sshaffer-at-microdataware.com MicroDataware http:www.microdataware.com (Under reconstruction and temporarily out of service) Personal stuff: steve_shaffer-at-compuserve.com http://ourworld.compuserve.com/homepages/steve_shaffer/ **********************************************************
Keith Ryan wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear Microscopy Listers } } I have been tasked with coming up with something about using light microscopes without harming yourself! Not so funny, according to two of our } now-retired staff who counted plankton etc. for 20+ years. They both suffered neck/shoulder/back pain which eased after retirement. } } In the UK we have a specific, detailed set of Regulations concerning computer use, another unspecific set covers all workstations e.g. supermarket } checkouts. A lot of the computer Regs. might be transferred to microscope set-ups. } } I'd appreciate any info, anything to avoid re-inventing the wheel! } } Season's Greetings } } Keith Ryan } Plymouth Marine Lab., UK Dear Keith,
Part of the problem is that few microscopists "align themselves", when they align the microscope. An adjustable chair, with secondary adjustments for the backrest, plus "elbow pads" or armrests on the microscope will solve most of these problems. The microscope should be addressed at eye level to avoid hunching over or stretching to reach the eyepieces.
If you decide to draft formal regs, send me a note.
Best regards, Barbara Foster (longtime FRMS) Consortium President Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108-2838 USA PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com **************************************************** Microscopy/Microscopy Education America's first consortium of microscopy experts offering customized on-site training & applications solutions
Adriana P. M Rodriguez wrote: } } Dear Dr. Foster: } I just saw a message you posted on the microscopy list, and the phrase } below your address caught my attention. } I just wondered if our institution offers short courses in different } microcopy techniques, or only on-site training. } Thanks in advance for your time. } } Adriana Rodriguez } } } Best regards, } } Barbara Foster } } Consortium President } } Microscopy/Microscopy Education } } 53 Eton Street } } Springfield, MA 01108-2838 USA } } PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com } } **************************************************** } } Microscopy/Microscopy Education } } America's first consortium of microscopy experts offering } } customized on-site training & applications solutions } } } } } Adriana P. M. Rodriguez } Biotecnologia Vegetal, CENA/USP } Av. Centenario 303, Cx. Postal 96 } 13400-970, Piracicaba, SP, Brasil } phone: +55-19- 429-4694 } fax: +55-19- 429-4610 Dear Adriana,
We will be offering several courses during first quarter 1998: a. SPIE/Photonics West - "Modern Microscopy & Applications" - lecture deomstrationwhich reviews fundamentals of microscopy, including a range of contrast enhancement techniques, plus a discussion of advanced methods including integration of image processing technis, automated measurement, and feed-back systems. (Thursday, 1/29/98, San Jose, CA)
b. American Chemical Society - "Applied Optical Microscopy for Chemists" - a three day, hands-on, total immersion course in light microscopy which covers imaging theory, contrast enhancement, and basic measurement techniques. Emphasizes Polatized Light Microscopy. (Friday, Saturday, Sunday, 2/27, 2/28, 3/1, New Orleans, LA)
c. The MME Traveling Road Show - "Optimizing Light Microscopy" - a lively, one-day lecture demonstration on alignment, operation, optics, and contrast enhancement. (Monday, 3/16, New York City; Wednesday, 3/18, Springfield, MA/Hartford CT; Friday, 3/18, Boston, MA).
For further information, please email me directly.
Thanks for your interest. Barbara Foster Consortium President Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108-2838 USA PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com **************************************************** Microscopy/Microscopy Education America's first consortium of microscopy experts offering customized on-site training & applications solutions
I just tried printing from ThumbsPlus 3.0. You can print in batch mode, you are not restricted to just a catalog and you can print a bunch of other information including:
name (with or without directory path) file size date dimensions pixels and physical size resolution keywords (that you added to it in the database) annotation (that you added)
All of these are optional in the output.
Agreeing with others, it is a great program. You can also scan images into it and the thumbnails are automatically made. We use the network version and have a LAN database that we can all select and we can have local databases for our hard drives. We open the images by double clicking on the image, copy it, and paste it into a window application such as a wordprocessor or PowerPoint. It can also be used to open a variety of formats including Photoshop and convert them.
-Scott
Scott D. Walck PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of S.D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
Rick Felten 12/15/97 12:52 PM Does anyone know a way to print several image files in an automatic fashion that would include the file name on the image. I am using windows 95.
Please unsubscribe nsmith-at-csuhayward.edu Nancy R. Smith Director of Operations Microscope And Graphic Imaging Center California State University, Hayward Hayward, CA 94542 http://www.csuhayward.edu/SCI/sem
Has anyone use Softwindows 95 for a Silicon Graphics system with the Scion Image PC (PC version of NIH Image)? Also what has been your experiences with Softwindows 95?
Six months ago I said goodbye to the List and my job managing the Australian Museum's SEM lab. I'm now in Brussels starting a Ph.D. in Archaeology, but it's funny how things move in circles. I'm looking at 3D modelling of excavations, including Image Analysis in 3D, which is why I'm starting by asking those of you in confocal microscopy the following;
1) Can anyone help with an email address for H.J.G. Gundersen at the Stereological Research Laboratory in Denmark? I've tried Stereo-at-svcd.aau.dk but the message bounces.
2) Does anyone know of commercially available 3D IA programs? (I want to measure object sizes and object orientations)
3) Does anyone know of any Lists dedicated to 3D modelling?
Many thanks, Merry Christmas and may your New Year's "resolution" be small and stable.
Is anyone out there doing any EDX work with marine sediments - especially those in the vicinity of pipeline outfalls. if so, I'd like to communicate with you.
Please reply to my e-mail address rather than the listserver. Thanks
Mike Gregory
EM Unit University of Durban-Westville South Africa
} Does anyone have experience using the Polaroid Sprint Scan 45 for TEM } negatives? I would appreciate any opinions on this scanner.
My response:
We have been using the SprintScan 45 for 6 months. We are very pleased with the quality of images produced by this scanner. Initially, an inconvenience of the scanner was the lack of a holder for TEM negatives. Polaroid has subsequently developed one that uses anti-Newton glass that works fine. I was very skeptical about the use of glass but it has proven to work nicely. The design of the holder is such that it can be easily modified with frames to make a glassless holder if you must have it. I suspect that these frames are in the works.
I have just completed (last night, in fact) a side by side comparison of images generated conventionally by darkroom endeavors versus scanning of the same negative and printing on a Codonics 1660 dye sub printer. Even though I had to use a color paper on the Codonics (their thermal line paper is not yet ready for prime time), the results were amazingly good. I compared enlargements of 1x, 4x, 8x and 16x using a 10MB image scanned at 2000 ppi. (I am now looking at smaller sized files as well.) I must say that as a "classically trained microscopist" with considerable dark room experience, I was finally convinced that a reasonable, high quality image can be generated in this way. I spent a couple hours in the darkroom using various focal length lenses, condensor lenses and adjustments on the enlarger to achieve the results that could have been obtained in 20 minutes on the scanner and dye sub printer. Although I think that a darkroom may be needed in some cases (large central facilities, for example), digital darkrooms will work for the majority of users. So fast, so convenient, so much less environmentally damaging.....
Disclaimer: I have no commercial interests in either Polaroid or Codonics. I can be just as critical of these companies when they show shortcomings.
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy Neckers Building, Room 146 - B Wing 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 ####################################################################
Hoping someone can help me. We used to buy PBS tablets from a company called OXOID. Need to order more but can't find address. Are they still around, or has someone bought them out. Any help greatly appreciated.
Thanks Mark Elliott UBC-Pulmonary Research Lab St. Paul's Hospital Vancouver, BC Canada V6Z 1Y6 604-631-5351 (fAX)
This is that magical time of the year when the thoughts of those of us in the tropics turn to... cryofixation!
We have a "cryogen freezing bath" device from Balzers, which consists of a small container on a tube which fits into a Dewar of LN2. The small container is filled with e.g., a freezing slush of Freon or propane, into which (cryoprotected) samples are plunged. Someone else on campus would like to have one of these devices, and the machine shops are really backed up. Do any of you have an idea where this kind of thing can be purchased? If you can't picture this, a schematic can be found in Bozzola and Russell's Electron Microscopy on p. 312.
Actually, while I'm at it, the intended purpose is to freeze mouse heart/aorta and liver for cryosections, and I'd be glad to pass on any tips any of you have. This group was just plunging the tissues into straight LN2 and wondering why they had big holes...! Pre-fixation is not a problem; in fact it is desirable. I think it's intended for in situ hybridization/autoradiography at the light level. It's those pesky photons, again. I just don't understand how they work, and so I'm not much help when it comes to LM!
It's about 76 degrees F and sunny in Honolulu today. Surf is coming up on the North Shore. Hope you all are having a fine holiday season!
Mele Kalkimaka, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
I am also very interesting in 3D image analysis and reconstruction, especially to the biological subjects. I send you an address in web site which contains a great number of software. Hopefully you could find some of them are suitable to your current project.
http://biocomp.arc.nasa.gov/3dreconstruction.
Good luck!
Peiyi Wang Research Fellow Department of Engineering Materials University of Southampton Southampton SO17 1BJ UK Tel: 01703 595101 Fax: 01703 593016 E-mail: pw2-at-soton.ac.uk
There is no real need to purchase the quench cooling device you describe. All you need is a copper cylinder which will sit in a Dewer of LN2. Have a little plastic lid on the cyliner, When the cyliner is cool, slowly condense some propane into the cold cylinder (do this in a spark proof hood). Once the propane is cooled put the lid on. The purpose of this little lid is to stop LN2 bubbling into the cylinder with the propane. When you are ready to quench your sample, get a warm rid and make a little pool in the solid propane, watch the propane pool and as the surface just begins to frreeze over, plunge in little bits of your sample. Let then stay for 30 secs then quickly bring them out and put them into a dewer of LN2. Variations of this are endlezs. You can use ethane instead of propane (bit dangerous) and you can have the whole dewer-cylinder sitting on a magnetic stirrer with a magnetic flea in the secondary cryogen. If you are really serios, read my book "Low temperature Microscopy and Analysis"
Your weather sounds great. Here in Cambridge we are just recovering from the Siberian Express which gave a wind chill factor of -10C. Now it hovers around freezing with rain. But we English argue that you have to suffer cold and wet to appreciAte warm
Best wishes for the Christmas Season
Patrick Echlin Multi-Imaging Centre School of Biological Science University of Cambridge United Kingom.
On Wed, 17 Dec 1997, Tina Carvalho wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } This is that magical time of the year when the thoughts of those of us in } the tropics turn to... cryofixation! } } We have a "cryogen freezing bath" device from Balzers, which consists of a } small container on a tube which fits into a Dewar of LN2. The small } container is filled with e.g., a freezing slush of Freon or propane, into } which (cryoprotected) samples are plunged. Someone else on campus would } like to have one of these devices, and the machine shops are really backed } up. Do any of you have an idea where this kind of thing can be purchased? } If you can't picture this, a schematic can be found in Bozzola and } Russell's Electron Microscopy on p. 312. } } Actually, while I'm at it, the intended purpose is to freeze mouse } heart/aorta and liver for cryosections, and I'd be glad to pass on any } tips any of you have. This group was just plunging the tissues into } straight LN2 and wondering why they had big holes...! Pre-fixation is not } a problem; in fact it is desirable. I think it's intended for in situ } hybridization/autoradiography at the light level. It's those pesky } photons, again. I just don't understand how they work, and so I'm not } much help when it comes to LM! } } It's about 76 degrees F and sunny in Honolulu today. Surf is coming up on } the North Shore. Hope you all are having a fine holiday season! } } Mele Kalkimaka, } Tina } } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } **************************************************************************** } }
Keith I assume that COSHH assessments will have taken care of fixatives, stains and immersion oil and that you aren't talking the risks of microtomes, knives and cryo work.
My feeling is that any assessment must look at levels of use:
If microscopes are used once a week for half an hour, keep them covered or in a cupboard and use them however you wish.
If microscopes are used regularly (like designated DSE user work) then a clear bench area in an a quiet area will be less stressful and ideally the user should be able to adjust the ambient light without disturbing others. Good seating should be provided which is adjustable or at least a variety of stools and chairs to suit all types of users also necessary basics like a stable bench surface, appropriate and well-placed power outlets (ie no trailing wires), lens tissues (keep those lenses clean), and a manual if necessary. Binocular eyepieces are fairly universal now and most users find these less stressful than monocular for prolonged use but they must know how to adjust them properly and should know how to set up the rest of the microscope for optimum use. Some spectacle users prefer to have high-viewpoint eyepieces so they can wear their glasses and some people prefer eyepiece eye cups (removable types are generally available). Adjustments such as X-Y stage controls, focus and lights should work properly (ie regularly maintained to avoid long term stress to the user). It may well be sensible to encourage people to take activity breaks as for DSE users. I would assume that if monitors are used to view the image they should be as suitable as modern computer screens ie sharp, no flicker and so on (if they are connected to computers then the DSE regulations should apply anyway).
Malcolm Haswell e.m. unit University of Sunderland UK
PS DSE is Display Screen Equipment, VDUs, or computer screens.
DISCLAIMER - these are my own personal opinions based on practical experience, even if I can't always achieve them.
----------
Dear Microscopy Listers
I have been tasked with coming up with something about using light microscopes without harming yourself! Not so funny, according to two of our now-retired staff who counted plankton etc. for 20+ years. They both suffered neck/shoulder/back pain which eased after retirement.
In the UK we have a specific, detailed set of Regulations concerning computer use, another unspecific set covers all workstations e.g. supermarket checkouts. A lot of the computer Regs. might be transferred to microscope set-ups.
I'd appreciate any info, anything to avoid re-inventing the wheel!
I am certain that this has been discussed before but can anyone give me guidance on the availability of large format black and white negative scanners for e.m. cut film (up to about 8.3 cm x 10.2 cm)?
I know that some flat-bed print scanners can take adaptors for film but it is not always obvious what format they can accommodate. At present the resolution of print scanners appears to be improving (600 dpi true resolution is not uncommon) and I suspect that for normal enlargement of 2 to 3x these would be just about adequate.
Any comments?
thanks Malcolm Haswell e.m. unit University of Sunderland UK
Oh and Merry Christmas to all my readers. ----------
Text item:
Does anyone have experience using the Polaroid Sprint Scan 45 for TEM negatives? I would appreciate any opinions on this scanner.
Disclaimer: The CSIR exercises no editorial control over E-mail messages originating in the organisation and the views in this message are therefore not necessarily those of the CSIR and/or its employees. Message-Id: {s4993452.076-at-csir.co.za} X-Mailer: Novell GroupWise 4.1
Hi We have a Jeol JSEM 200 STEM (200 kV) available at a nominal cost for anybody interested. If you are interested in the machine, or can want a part (high voltage transformer, lenses, scanning system, specimen holders, gun, etc), please let me know as soon as possible as the machine is heading for the scrapyard... All shipping must be paid by yourself.
Sara Prins
Surface and Structure Analytical Services Division for Materials Science and Technology CSIR PO Box 395 Pretoria South Africa
} There is no real need to purchase the quench cooling device you } describe.... a little pool in the solid propane..... Variations of this } are endless.
ISO-pentane freezes at about -160^C and comes in a bottle.
+------------------------------------------------------------------------+ | 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 | +------------------------------------------------------------------------+
We are interested in imaging, (non-destructively) the internal morphology of 6 - 8 micron polymeric particles. The SEM and TEM have already been used for this study and we have determined the size of internal voids to be between 0.1 and 0.4 microns. Does anyone know the resolution limits of acoustic microscopy and the location of a facility where I might get such work done?
Gerroir,Paul wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } We are interested in imaging, (non-destructively) the internal } morphology of 6 - 8 micron polymeric particles. The SEM and TEM have } already been used for this study and we have determined the size of } internal voids to be between 0.1 and 0.4 microns. Does anyone know the } resolution limits of acoustic microscopy and the location of a } facility where I might get such work done? } } Paul Gerroir Dear Paul,
I'd suggest to talk to the people at SONOSCAN and Leica. Both have acoustical microscopes in their product lines and may be able to suggest a facility close to yours.
My contact at Sonoscan is Don Commare (708/766-7088). I'm not sure who the Leica product manager is but you can reach Leica at (847)405-0123; they can put you in touch with the correct person.
Best of luck.
Barbara Foster Consortium President Microscopy/Microscopy Education 53 Eton Street Springfield, MA 01108-2838 USA PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com **************************************************** Microscopy/Microscopy Education America's first consortium of microscopy experts offering customized on-site training & applications solutions
Am I missing something ? I never mentioned Isopentane.Of the two I mentioned ETHANE melting point 90K gives a mean cooling rate of 13-15 kKsec-1, PROPANE melting point 84K gives a mean cooling rate of 10-12 kKsec-1. Your stuff, ISOPENTANE melting point 113K does not cool as fast as the other two although I do not have the figures to hand. If you want to read more see Chapter 3 in "Low Temperature Microscopy and Analysis" by Patrick Echlin, Plenum Press, New York 1992. When choosing a cryogen there are many factors to consider including specific heat, thermal conductivity, relative cooling efficiency, viscosity at the crogen melting point andthermal inertia.
To all cryomicroscopists the Seasons Greetings.
Patrick Echlin Cambridge
Thu, 18 Dec 1997, Robert H. Olley wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } } On Thu, 18 Dec 1997, Dr P. Echlin wrote: } } } There is no real need to purchase the quench cooling device you } } describe.... a little pool in the solid propane..... Variations of this } } are endless. } } ISO-pentane freezes at about -160^C and comes in a bottle. } } +------------------------------------------------------------------------+ } | 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 | } +------------------------------------------------------------------------+ } }
We have small and portable "Hanna HI8424 microcomputer pH meter" with an electrode that now reads about ph=8.1 instead of pH=7.0 the true value. When I push the calibrate button though, it just reads error (E4), and there is no way that I can calibrate this meter.
The electrode only says HI 1332 on it, and it appears to be a Ag/AgCl electrode. My problem is that I don't see "Hanna" listed in any catalogs, so I'm not sure what electrode I should buy to replace this electrode. It connects to the main control unit with a BNC connector. I have already tried rejuvinating this electrode by soaking it in various solutions, but so far nothing has changed in the way it reads.
Are there any Hanna pH meter types out there that might be able to advise me on what electrode I should get, and where I can get this electrode?
Paul Gerroir wrote: ========================================= We are interested in imaging, (non-destructively) the internal morphology of 6 - 8 micron polymeric particles. The SEM and TEM have already been used for this study and we have determined the size of internal voids to be between 0.1 and 0.4 microns. Does anyone know the resolution limits of acoustic microscopy and the location of a facility where I might get such work done? ============================================ You might want to contact a firm in suburban Chicago called Sonoscan, Inc. as follows:
Sonoscan, Inc. 530 E. Green St. Bensenville, IL 60106 USA
Tel: 630-766-7088 Fax: 630-766-4603
You would want to talk to Dr. Larry Kessler who is the founder and, so far as I know, still the President. He has helped me out of any number of tricky problems that were solvable with or only with accoustic microscopy. He is very knowledge about material science applications of accoustic microscopy. But to my understand, which might not be state-of-the-art, the resolution you are seeking I don't think is possible even with accoustic microscopy.
Chuck
Disclaimer: I have no connection with Sonoscan, financial or other wise.
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Dear Judy, Scott, and David: Thank you for your response to my question on TEM calibration. I am following up on your suggestions.
Augusto _________________________________________________________________________ Augusto A. Morrone 107D-MEL, P.O. Box 116400 MAIC Materials Science and Engineering University of Florida Gainesville, FL 32611 (352) 392-1497 or 6985 Fax: (352) 392-0390 amorr-at-mse.ufl.edu
See our Web Page for detailed information about our current opening for an engineering manager. web site:kovexcorp.com or e-mail me your resume : kovex-at-spacestar.net, or telephone 612-486-9830 ext 113.
I have some investigators that are interested in doing shadow casting of DNA and large protien structures, but we do not have a carbon evaporator. They would like me to get some sources and prices for a equipment grant. Other than Electron Microscopy Sciences who else sells them? Are there any opinions on this matter? Thanks.
Rick Vaughn
RLVAUGHN-at-MAIL.UNMC.EDU EM Research Facility Dept. Cell Biology & Anatomy UNMC Omaha, NE
I have been asked to find a source for Fast Blue B.
According to the person making the request, Sigma, Fisher, and Acros have all discontinued all Fast Blue B Salts and precursors.
Does anyone know of a source or have a stockpile on hand?
According to the requester, Fast Blue B and its salts are carcinogenic and this is the reason they are no longer available. Don't ask me why he wants to fool around with something like this, I am just the messenger.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
Dear Friends, I had the opportunity to share someone's: YOUR collegial friendship in = communicating more or less frequently via the MSA-Listserver or on else = opportunity. I received most valuable informations and help regarding my = requests (some were urgent) and will forward as intensely my knowledge, = if appropriate. For all those I promised informations, reprints, methods, silicone = rubber molds and instructions and else material: please apologize at = that moment for my being late with a lot of things. The month before = Christmas time every year is a strong one regarding daily stress of work = (all specimen preparations in our Lab. on my own, lectures, official and = administrative work, etc....and singing in a choir, the SALZBURG = MOZART-CHOIR with 4 different concert performances this month). I didn't forget (hope so) any of you and of my promises to you. Please allow for another some weeks, hopefully after holidays things = will go on faster than now. If one feels that something was not received = or mailed which should have been in the mail for a long time, please = feel free to urge or claim.
To all of you and yours greetings and my best wishes=20 for A Merry Christmas, at least some calm, familial and quiet days of = relaxation, as well as all the best for a HEALTHY, HAPPY, PROSPEROUS and = SUCCESSFUL NEW YEAR.
yours truly, *Wolfgang*
(Out of the Lab from 23rd Dec. to 7th of Jan, '98; but as I know me, I = will be in at some days in between for "work-up" of unsettled items)
Dr. Wolfgang MUSS Department of Pathology, LKA EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
SALZBURG, 19th of Dec. 1997, 01.35 a.m. local time
Dear Garry, dear all, surprising doublette:=20 the same happens/-ed with my pH-meter (BECKMAN ZEROMATIC IV) or better = the connected combination electrode (Ag/AgCl). I confess, the meter/electrode was not in use for three or four months = now, but: electrode was soaked all the time in a so called "storing = solution" originally supplied from BECKMAN (solution adjusted to pH = 4.0). I am watching now about 4 weeks what happens: despite "stand = by-function", the analog meter showed initially slowly increasing values = starting pH 7.0 to 8.5, now, after 4 weeks, it "displays" (without being = "in action") pH 11.0. If changing function from "Stand By"-mode to = "measuring" it displays pH 8.0. Now idea what goes on. Maybe a malfunction of an electrode too old = aged?? (about 5 years of age). I have not tried to compensate or to = refresh the electrode anyway, but there is white crystal growth on the = upper outside of the electrode tube, far away from the solutions in the = storage baker. Somebody out there with an idea? (BECKMAN as "last chance", I know)
Thanks in advance, best regards,
Dr. Wolfgang MUSS Department of Pathology, LKA EM-Laboratory Muellner Hauptstrasse 48 A-5020 SALZBURG AUSTRIA/Europe
phone: ++43++ 662 + 4482 + 4720 Ext fax: ++43++ 662 + 4482 + 882 Ext. e-mail: W.Muss-at-lkasbg.gv.at (note: "l" right to "-at-" is a small "L")
=20 GARRY BURGESS WROTE: ---------- Von: Garry Burgess[SMTP:GBurgess-at-exchange.hsc.mb.ca] Gesendet: Donnerstag, 18. Dezember 1997 20:49 An: 'Microscopy Society of America - Mailing List' Betreff: Hanna HI8424 pH Meter
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20
We have small and portable "Hanna HI8424 microcomputer pH meter" with an electrode that now reads about ph=3D8.1 instead of pH=3D7.0 the true = value. When I push the calibrate button though, it just reads error (E4), and there is no way that I can calibrate this meter.
The electrode only says HI 1332 on it, and it appears to be a Ag/AgCl electrode. My problem is that I don't see "Hanna" listed in any catalogs, so I'm not sure what electrode I should buy to replace this electrode. It connects to the main control unit with a BNC connector. I have already tried rejuvinating this electrode by soaking it in various solutions, but so far nothing has changed in the way it reads.
Are there any Hanna pH meter types out there that might be able to advise me on what electrode I should get, and where I can get this electrode?
Dr Echlin is quite right. Isopentane had been used as a freezing compound a long time ago, but I sure was glad when propane was advocated as a better alternative. I started using propane about 15 years ago. Isopentane behaves like a "gone off" rubber solution near its freezing point and freezes well before propane. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au
} On Thu, 18 Dec 1997, Dr P. Echlin wrote: } } } There is no real need to purchase the quench cooling device you } } describe.... a little pool in the solid propane..... Variations of this } } are endless. } } ISO-pentane freezes at about -160^C and comes in a bottle. } } +------------------------------------------------------------------------+ } | Robert H.Olley Phone: |
Malcolm, our Links page has two sites that deal with "scanners". Use control F to find these sites. The site at Basel Uni lists numerous scanners and gives a good deal of technical details. Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au
} } I am certain that this has been discussed before but can anyone give me } guidance on the availability of large format black and white negative } scanners for e.m. cut film (up to about 8.3 cm x 10.2 cm)? } } I know that some flat-bed print scanners can take adaptors for film but it } is not always obvious what format they can accommodate. At present the } resolution of print scanners appears to be improving (600 dpi true } resolution is not uncommon) and I suspect that for normal enlargement of 2 } to 3x these would be just about adequate. } } Any comments? } } thanks } Malcolm Haswell } e.m. unit } University of Sunderland } UK } } Oh and Merry Christmas to all my readers. } ---------- } From: Leah L Dobbs } To: Microscopy-request; microscopy } Subject: Negative Scanners } Date: 17 December 1997 17:51 } } Text item: } } Does anyone have experience using the Polaroid Sprint Scan 45 for TEM } negatives? I would appreciate any opinions on this scanner. } } Thanks, } } Leah L Dobbs } TEM Analyst } Intel Corporation }
} Dr Echlin is quite right. Isopentane had been used as a freezing compound a } long time ago, but I sure was glad when propane was advocated as a better } alternative. I started using propane about 15 years ago. Isopentane behaves } like a "gone off" rubber solution near its freezing point and freezes well } before propane.
Jim Darley is also right. I had overestimated iso-pentane, partly as a result of getting my Celsius and Kelvin mixed up! But for less critical work, like rapid quenching of polymer melts, it is sometimes helpful where working with a bottle of gas might not be allowed.
+------------------------------------------------------------------------+ | 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've got a Hanna HI 8520 with autocalibrate at 4.01, 7.0 and 10.0. It is new and works fine at the moment with it's supplied electrode but will not autocalibrate with an TRIS resistant electrode that we bought at the same time. I still need to contact my suppliers. I will let you know if I get anywhere.
Malcolm Haswell University of Sunderland UK ----------
Ricky L Vaughn wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } I have some investigators that are interested in doing shadow casting of } DNA and large protien structures, but we do not have a carbon } evaporator. They would like me to get some sources and prices for a } equipment grant. Other than Electron Microscopy Sciences who else } sells them? Are there any opinions on this matter? Thanks. } } Rick Vaughn } } RLVAUGHN-at-MAIL.UNMC.EDU } EM Research Facility } Dept. Cell Biology & Anatomy } UNMC } Omaha, NE
Rick,
We at Ladd Research are among those who manufracture and sell carbon evaporation systems. Please e-mail or call 1-800-451-3406 and we can talk options and pricing.
John Arnott Ladd Research 13 Dorset Lane Williston, VT 05495
TEL (US) 1-800-451-6406 1-802-878-6711 FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net
Hi Fellow Microscopists, We have just installed a "windowless" Si(Li) EDS detector for our ESEM. We would like to establish the spatial resolution of the detector in the system as a function of our typical ESEM conditions: 15KV, short working distance using the extended bullet, about 1 torr pressure, gas: air, at room and cryogenic temperatures. We are curious if fellow ESEMer's have undertaken a similar study and have some numbers to share. Are there better conditions to optimize performance? Any suggestions for good standards to test the spatial resolution?
HOPE YOUR HOLIDAYS ARE MAGNIFI-CENT!!
Lynne Garone Robert Baron Polaroid Corp. GaroneL-at-Polaroid.com
Can anyone please tell me the "a" spacing of 302 St. Steel, using the JEOL 100CX TEM/SCAN viewed at 100kv.....or how i could find that out?........Please email me ASAP Thank You.....and have a Merry Christmas Brandon Hernandez (EM student)
O.k., here's a list to get you started. I know I've left some off - no offense was indended, and hopefully the vendors themselves contact you.
NOTE: One strong recommendation. Apparently most vacuum evaporator companies supply for the optical and elctronics industries, NOT MICROSCOPY, make sure when you order that if the system has an Oil diffusion pump you specify that it NOT be filled with Silicon oil (Silicon + EM = VERY BAD), but a non-Si based oil (i.e. Santovac, or alternative).
Carbon Evaporator Vendors: (Alphbetically)
Balzers
[North American Rep.] Technotrade International 7 Perimeter Road Manchester, NH 03103-3343 Tel (603)622-5011 Fax (603)622-5211
=} Balzers is primarily involved with vacuum coating systems for the optical industry, but decades ago (1960's?) expanded their systems into the microscopy world as well. Tend to be (1) expensive, (2) finicky, (3) Techno Trade is very helpful to deal with though.
Denton Vacuum Inc. 1259 North Church Street Moorestown, NJ 08057
PH: 609.439.9100 FAX: 609.439.9111
=} I presently have two different models and they work quite nicely. The design / layout of electrical connectors, mechanical connetor (rotator) could be better for ease (Need two wrenches) and keeping clean. but not enough to swear off of them.
Edwards High Vacuum International 301 Ballardvale Street Wilmington, MA 01887
Ladd Research Industries, Inc. P.O. Box 1005 Burlington, VT 05402 PH: 802.878.6711 FAX: 802.878.8074
=} Bill Ladd designed one of the best, easiest to configure, easiest to clean, most flexible stage areas I have ever worked with (My Opinion). But Bill left Ladd a few years ago, and things were ... fuzy (?) for a while. Haven't dealt with them in three years.
Ted Pella, Inc. 4595 Mountain Lakes Blvd. Redding, CA 96003
Albert Prebus, one of the pioneers of transmission electron microscopy, died at his home in Columbus (Ohio) last Tuesday. The funeral will take place today (Friday).
After receiving B.Sc and MSc. degrees from the University of Alberta, he constructed the first North-American electron microscope at the University of Toronto in 1938, for which he earned the first Ph.D granted in electron microscopy. His achievments were honored in 1978 with an honorary Doctor of Science degree from the University of Toronto, presented at the 9th International Congress on Electron Microscopy.
In 1940, Dr. Prebus joined the Department of Physics at Ohio State University and retired as Professor Emeritus in 1978.
Ray Egerton, Physics Dept, University of Alberta, Edmonton, Canada T6G 2J1 Phone: 403-492-5095, FAX: 403-492-0714, e-mail: egerton-at-phys.ualberta.ca ------------------------------------------------------------------------
Dear Rick Vaughn: Two worthy carbon evaporator manufacturers come to mind: Edwards High Vacuum at 1-800-848-9800 and Denton Vacuum at 609-439-9100. Our Denton 502A evaporator is in its 13th year and has provided very reliable service and when technical information was needed they are very helpful people. I suppose this is a shameless plug for a company with which I have no financial interest. Good luck in your search. Don Gantz Biophysics Department Boston Univ Medical School
O.k., all you digital imaging folks: Looking for comments of any kind comparing the Condonics NP-1600 and the Tektronix Phaser 450. We're presently in the market to purchase a full page "publication quality" dye-sublimation printer, and based on vendor lit., listserver comments, and industry reviews. (Where as the Fuji Pictography 3000 looks really nice its just a little out of our price range, scarily slow).
Image Sources:
- Need to print via a networked solution, perferably a directly networked printer.
- Printing variety of images, but primarily Greyscale EM, color Confocal, and full color LM images. Ranging from 256 x 256, on up to over 5,000 x 5,000. (Fully realizing that at 300dpi 1:1 isn't going to happen)
- Printing from Intel PC's (Primarily Win NT) and Some Mac's, and a variety of software.
Codonics:
Whereas this printer comes highly regommended by the Microscopy community (Even recommened by a vendor who sold the Tektronix and NOT the Condonics) I have some grave concerns about it.
(1) Considering the highly computer oriented nature of digital imaging, the Condonics web site(http:\\www.codonics.com\) is pathetic, and last updated Aug, 6, 1997 with the NP-1600 page updated Dec, 5, 1995!). No online tech support, no on-line drivers, no FAQ's, no even a downloadable PDF manual. Whereas the "built in floppy drive for easy upgradability" maybe great but where do the "upgrades" come from? Snail mail communications? Any feeling for longer term support? (Last Codonics I worked with, '90-'94, worked great, but 1.5 years after purchase Tech support didn't really want to help at all with drivers for Windows 3.1 world, they had moved on to better things I guess)
(2) What, if any, options are there?
(3) Web search for "codonics" only results in 50-60 different sites for "Instructions for printing to the Codonics printer".
(4) No OS specific drivers, simply allows straight transfer of most image formats to the printer (this is nice) or relies on Post-Script printing (which is an option? How is it implemented?), but does require "loging on" and some rather criptic numerical "print-like-this" mode commands. I take it you are stuck with 1:1 printing and thus have to scale your image sizes prior to printing.
(5) From the user end (since most users aren't computer techno geeks like some of the rest of us) how user friendly is it really?
(6) Very fast, to Fastest on market - great. Any comments, particularly compared to Tektronix?
(7) Handles multiple jobs simultaneously - again great! Any comments?
TEKTRONIX
(1) Solid support for a variety of printing environs but primarily graphics industry, not sci. imaging.
(2) Solid on-line tech support.
(3) Requires OS specific drivers - how easily installed?
(4) Requires memory upgrade for larger image printing.
Any requested confidentiality for candid comments will be strongly protected. Vendors should feel free to reply as well.
Thank you.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"640K ought to be enough for anybody." -- Bill Gates, 1981
} We would like to establish the spatial resolution of the } detector in the system as a function of our typical ESEM conditions: } 15KV, short working distance using the extended bullet, about 1 torr } pressure, gas: air, at room and cryogenic temperatures.
David Joy has a Monte Carlo program to calculate the emission volume for x-rays. I don't know whether he has anything which will account for the E in the ESEM, but I'd ask him (the beam spread and x-ray absorption might be negligable anyway).
} Any suggestions for good standards to test the spatial resolution? } That depends on what you are going to examine. The SR depends on the average Z of the matrix among other things, and it can also depend on the element of interest. If you will be looking at biological specimens, perhaps something like magnetotactic bacteria would be useful--they have ~1 mu-m particles of magnetite. If you have some small particles of the element of interest, you could disperse them in a sucrose solution. (I calculated the % sucrose which gives about the right density and compo- sition for cells, but I don't have it with me.) I wonder if MAG*I*CAL can be useful for mineral specimens. You might be able to detect the Ge in the Si matrix, but I don't remember whether the Ge-containing bits are separated by enough distance. I have measured SR (crudely) by taking spectra on particles and in the matrix near them. This could be done correctly to get some function similar to a point spread function (of course, there is not enough signal from a point, so you'll have to examine an area { { your probe). Good luck. Yours, Bill Tivol
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
O.k., all you digital imaging folks: Looking for comments of any kind comparing the Condonics NP-1600 and the Tektronix Phaser 450. We're presently in the market to purchase a full page "publication quality" dye-sublimation printer, and based on vendor lit., listserver comments, and industry reviews. (Where as the Fuji Pictography 3000 looks really nice its just a little out of our price range, scarily slow).
Image Sources:
- Need to print via a networked solution, perferably a directly networked printer.
- Printing variety of images, but primarily Greyscale EM, color Confocal, and full color LM images. Ranging from 256 x 256, on up to over 5,000 x 5,000. (Fully realizing that at 300dpi 1:1 isn't going to happen)
- Printing from Intel PC's (Primarily Win NT) and Some Mac's, and a variety of software.
Codonics:
Whereas this printer comes highly regommended by the Microscopy community (Even recommened by a vendor who sold the Tektronix and NOT the Condonics) I have some grave concerns about it.
(1) Considering the highly computer oriented nature of digital imaging, the Condonics web site(http:\\www.codonics.com\) is pathetic, and last updated Aug, 6, 1997 with the NP-1600 page updated Dec, 5, 1995!). No online tech support, no on-line drivers, no FAQ's, no even a downloadable PDF manual. Whereas the "built in floppy drive for easy upgradability" maybe great but where do the "upgrades" come from? Snail mail communications? Any feeling for longer term support? (Last Codonics I worked with, '90-'94, worked great, but 1.5 years after purchase Tech support didn't really want to help at all with drivers for Windows 3.1 world, they had moved on to better things I guess)
(2) What, if any, options are there?
(3) Web search for "codonics" only results in 50-60 different sites for "Instructions for printing to the Codonics printer".
(4) No OS specific drivers, simply allows straight transfer of most image formats to the printer (this is nice) or relies on Post-Script printing (which is an option? How is it implemented?), but does require "loging on" and some rather criptic numerical "print-like-this" mode commands. I take it you are stuck with 1:1 printing and thus have to scale your image sizes prior to printing.
(5) From the user end (since most users aren't computer techno geeks like some of the rest of us) how user friendly is it really?
(6) Very fast, to Fastest on market - great. Any comments, particularly compared to Tektronix?
(7) Handles multiple jobs simultaneously - again great! Any comments?
TEKTRONIX
(1) Solid support for a variety of printing environs but primarily graphics industry, not sci. imaging.
(2) Solid on-line tech support.
(3) Requires OS specific drivers - how easily installed?
(4) Requires memory upgrade for larger image printing.
Any requested confidentiality for candid comments will be strongly protected. Vendors should feel free to reply as well.
Thank you.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
"640K ought to be enough for anybody." -- Bill Gates, 1981
FEI Company in Hillsboro, OR has the following position open:
JOB TITLE: Particle Optics Scientist DEPARTMENT: Systems R&D REPORTS TO: R&D Manager
1) GENERAL PURPOSE:
Perform particle optics experiments and provide assistance for development of new particle optics devices.
2) ESSENTIAL RESPONSIBILITIES:
A) Perform particle optics calculations on electron / ion columns or analyzers using Munro and / or similar programs. These can include magnetic and electrostatic fields, deflection systems, as well as paraxial, 2D or full 3D modeling. Beam interactions can also be involved. B) Perform experiments on new particle optical devices. C) Keep up with the particle optics community through the literature and conferences. D) Write internal specifications, proposals and reports. E) Interact with customers regarding requirements for new particle optics. F) Serve on new product development teams as scientific advisor and for testing.
3) EDUCATION AND/OR EXPERIENCE:
A) Ph.D. is science or engineering. B) 4 years experience using Munro or similar particle optics programs. C) Particle optics experimental experience. D) Experience installing and maintaining particle optic programs on a PC. E) Demonstrated creativity. F) Ability to modify existing or write special particle optics programs.
Interested applicants should submit their resume to Lisa Olivia either by FAX at 503-640-7509 or email at lolivia-at-feico.com FEI Company, 7451 N.W. Evergreen Parkway, Hillsboro, OR 97124 ****
FEI Company in Hillsboro, OR has the following position available:
JOB TITLE: DualBeam Applications Development Engineer DEPARTMENT: Applications Development REPORTS TO: Applications Development Manager
SUPERVISORY RESPONSIBILITY:
Generally none, but may give general direction to various laboratory assistants and technicians.
JOB SUMMARY:
Develop applications for the DualBeam market, including demonstration and sales support as necessary.
ESSENTIAL RESPONSIBILITIES:
1. Conceive, plan and conduct experimental research to advance applications knowledge as applied to use of DualBeam in-line and laboratory FIB/SEM tools. 2. Acquire information about customer requirements through observation of FEI customer FIB demos, direct contact with customers and from other available marketing input. 3. Aid specification of software and hardware development or modification as necessary to implement applications consistent with safety considerations. 4. Recommend and advise on direction for potential DualBeam applications consistent with customer requirements and technical feasibility. 5. As necessary, demonstrate system operation and applications in support of Technical Marketing efforts. 6. Develop and conduct customer training programs in DualBeam applications either on site at FEI or at customer facility. 7. Develop and conduct technology transfer documents and training programs in DualBeam applications for FEI technical staff. 8. Develop scientific and technical information at conferences and meetings in support of sales and marketing efforts. 9. Disseminate general information regarding technologies relevant to advancement of DualBeam applications.
OTHER DUTIES:
1. Provide scientific and technical support to R & D, Manufacturing, Customer Service and other departments as required. 2. Assist with writing manuals and reviews. 3. Other duties as temporarily assigned by supervisor.
MINIMUM EDUCATION, EXPERIENCE AND QUALIFICATIONS:
1. MS in Physics, Chemistry or related discipline. 2. 2+ years experience in operating FIB, DualBeam=99, SEM or similar analytical systems in a lab or FAB environment. 3. Experience in defect review, inspection and other semiconductor process applications is highly desired. 4. Able to interact effectively with various employees, customers and groups at FEI and customer sites. 5. Familiar with UHV 6. Willing and able to travel up to 20% of time. 7.Eligible for passport.
Interested candidates should submit their resume to Lisa Olivia, preferably by email: lolivia-at-feico.com or FAX: 503-640-7509 FEI Company, 7451 N.W. Evergreen Parkway, Hillsboro, OR 97124
Atomic Structure / Property Relations of Interfaces in TiAl
An opening exists for a postdoctoral fellow in my research group at the University of Pennsylvania. This position is funded by a grant from the National Science Foundation on the study of interface atomic structure and diffusion in a model material system. The work will be primarily experimental with heavy emphasis on the use of state-of-the-art electron microscopy facilities at Penn. Characterization of the atomic structure and composition profile of interfaces will be correlated with measurements of interface diffusion by in-situ experiments. The in-situ experiments will be conducted at Penn and at Oak Ridge National Laboratory through an ongoing formalized collaboration. The post-doctoral fellow will likely make multiple trips to ORNL in connection with this work. The project will also include continuous collaboration with another Penn group conducting an atomistic modeling study of interface diffusion. Experimental facilities at Penn critical to this effort are a new JEOL 2010F field emission gun transmission electron microscope with several unique capabilities, a JEOL 4000 high-resolution transmission electron microscope, a Phi scanning Auger microprobe, and an optical float zone furnace. Materials Science at Penn has a wide range of other experimental facilities, excellent on-site computational hardware, an active and highly regarded faculty and a vibrant research atmosphere. If you are interested in learning more about this challenging opportunity, contact me at luzzi-at-lrsm.upenn.edu.
David E. Luzzi Professor Department of Materials Science and Engineering University of Pennsylvania Philadelphia, PA 19104-6272
The former Botany Department at the University of California at Davis has two instruments available to interested folks.
The first instrument is a JEOL 100S TEM, vintage about 1981. If you are interested, contact Gary Zamzow (530-752-8865;gwzamzow-at-ucdavis.edu).
The second instrument is an Hitachi S-800 SEM, vintage about 1981. This is one of Hitachi's early FEG instruments and has had some modifications, it may also have a Kevex x-ray system. If you are interested, contact Rick Falk (rhfalk-at-ucdavis.edu).
I am a former graduate student in the former department and would be happy to tell you what I know about the instruments, but you must contact those listed above for information regarding price or other terms.
Jonathan Krupp Microscopy and Imaging Lab University of California Santa Cruz, CA 95064 (408) 459-2477 FAX (408) 429-0146 jmkrupp-at-cats.ucsc.edu
Thanks for all of the suggestions, I've passed on the information to Matt, and it looks like the Eppendorf finder slip is the best for his application. -- Randy Nessler rnessler-at-emiris.iaf.uiowa.edu Views expressed are my own.
Merry Christmas as well as all the best for Healthy, Happy, Prosperous and Successful New Year yours truly J o z e f Stankovic p.s. tento subor je nutne odkodovat programom MIME64 resp. BASE64 a az potom spustit, lebo inac nefunguje.
This message is in MIME format. Since your mail reader does not understand this format, some or all of this message may not be legible.
I remember seeing a reference to an Immunocytochemistry Network but did not save the address. Could someone please make that available. Thankyou. Don Gantz Biophysics Dept. Boston Univ Medical School gantz-at-med-biophd.bu.edu
Lynne You may want to reconsider your windowless detector for the ESEM. Windowless detectors on a high vacuum system are difficult to maintain because all the water, hydrocarbons, and junk in the vacuum ends up on the crystal because it is at liquid nitrogen temperature. The only way that I could see this working was if you only went windowless when in an inert atmosphere such as dry nitrogen, argon, or the like. You mention air at 1 torr at room temperature which seems incompatable with windowless EDS operation. I do have some data on operating conditions for analysis and how to test your xray resolution - please contact me off the list. Happy holidays, Scott Wight
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------------------------------ Scott Wight e-mail: SCOTT.WIGHT-at-NIST.GOV NIST - Microanalysis Group W voice: 301-975-3949 Bld 222, Rm A113 | fax:301-216-1134/301-417-1321 Gaithersburg, MD 20899 \|/ disclaimer: Any opinion expressed is my own and does not represent those of my employer.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
FYI, Small World has a ESEM version of the electron flight simulator which is based on David Joys code. I have no financial interest in Small World, I am just a beta tester of the ESEM version. Scott Wight
------------------------------------------------------------------ Scott Wight e-mail: SCOTT.WIGHT-at-NIST.GOV NIST - Microanalysis Group W voice: 301-975-3949 Bld 222, Rm A113 | fax:301-216-1134/301-417-1321 Gaithersburg, MD 20899 \|/ disclaimer: Any opinion expressed is my own and does not represent those of my employer.
edelmare-at-casmail.muohio.edu wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------.} } } } } Ladd Research Industries, Inc. } P.O. Box 1005 } Burlington, VT 05402 } PH: 802.878.6711 } FAX: 802.878.8074 } } =} Bill Ladd designed one of the best, easiest to configure, easiest } to clean, most flexible stage areas I have ever worked with (My } Opinion).
To: Richard Edelman, Ph.D. Miami University
Thank you for your kind words concerning the Ladd evaporator. There are many users in the microscope world that would agree with you that Bill Ladd's evaporator was "one of the best, easiest to configure, easiest to clean, most flexible stage areas" etc. Ladd continues not only to produce that evaporator but we have even improved it.
Both Margaret and Bill passed on a few years ago but their long-time employees continue their tradition of quallity today after more than 40 years. It is always difficult to follow pioneers such as Margaret and Bill, but we learned from them and know they'd be proud that we continue to supply the microscopy world with the same quality of products that they did for oh so many years.
Thank you for your kind words concerning the Ladd Evaporator. There are many users in the microscope world that would agree with you that Bill Ladd's evaporator was "one of the best, easiest to configure, easiest to clean, most flexible stage areas" etc. Ladd continues not only to produce that evaporator but we have even improved it.
Both Margaret and Bill passed on a few years ago but their long-time employees continue their tradition of quality today after more than 40 years. It is always difficult to follow pioneers such as Margaret and Bill, but we learned from them and know they'd be proud that we continue to supply the microscopy world with the same quality products they did for oh so many years.
It looks as if we are about to get a bunch of 'unsubscribe' for the holiday messages (sent to the list rather than the listserver for some reason). I'm sure there is a 'postpone' command that has a complementary 'unpostpone' command. These can be sent to the listserver any time you want cessation of mail delivery in the short term.
i) Am I correct? and ii) If so, can someone in the know tell us all the proper command phrase and the address to send it to (listserv-at-msa.microscopy.com?)
Ho-Ho-Ho, John
================= C. John Runions, Ph.D Section of Ecology and Systematics Corson Hall Cornell University Ithaca, New York USA 14853
I work for VayTek. We sell a volume rendering product called VoxBlast.
I'm responding to Geoff Avern's question on December 17th concerning 3D. } 2) Does anyone know of commercially available 3D IA programs? (I want to } measure object sizes and object orientations) } VoxBlast is a 3D volume visualisation and measurement software that runs on UNIX, Windows 95/NT, and Mac. VoxBlast creates a volume from a stack of 2D images. The 2D images can be from many sources such as bright field, fluorescence, confocal, MRI, PET, CT, geological, etc. VoxBlast also has measuremement tools to give you object size and coordinates for selected points in 3D space. Our Windows version currently has a 3D Object Counting function that gives you the XYZ coordinates, volume, average density of each object found fitting the specified cluster size limits, and gray level limits.
If you have any interest in VoxBlast, please go to our web site at WWW.VAYTEK.COM . You can take a look at comparative rendering times for many machines on all three platforms. You can also download demo versions for each of the platforms.
Best regards
Patrick Guerin Customer Technical Support Engineer VayTek, Inc. 305 West Lowe Avenue, Suite 109 PO Box 732 Fairfield Iowa 52556-0732 Tel : 1-515-472-2227 Fax : 1-515-472-8131 E-mail : pguerin-at-vaytek.com _______________________________________
I would need information on solutions used in order to dilute antibodies in immunohistochemistry
THANK YOU
---------------------------------------------------------- Dr. Hugo H. Ortega Laboratorio de Investigaciones Histologicas Aplicadas Catedra de Histologia y Embriologia Facultad de Agronomia y Veterinaria Universidad Nacional del Litoral
There is no postpone/unpostpone command, please review your instructions which you all received when you first subscribed. Or review them on the MSA WWW Site.
------------------------------------------------------ To remove yourself for the holidays
Simple send an UNSUBSCRIBE Email message to
Listserver-at-MSA.Microscopy.Com
then SUBSCRIBE once again when you return to
Listserver-at-MSA.Microscopy.Com
------------------------------------------------------ P.S. Please remember you include your subscription address for many people this may not be identical to your current email address if your mail is forwarded or you use an alias! ------------------------------------------------------
To avoid the mass of annual seansonal messages allow me to post a generic message:
*********************************************** Everyone on the Listserver wishes Everyone else a healthy and happy holiday season! ***********************************************
Cheers.... Nestor Your Friendly Neighborhood SysOp.
Our department is trying to find a way of making 3D reconstruction of plants under development. I am aware of something called "3D digitizer", but I am not sure how it works.
For the moment we are just making movies of plants and taking 2D images around the plant (36 images) such that we can make a QTVR film.
Maybe this is a subject for a 3D reconstruction mailing list, does somebody know of such a list?.
can anybody tell me where in Germany or Europe I can buy collodion (celloidin, parlodion), as a powder or as a solution in amylacetate. I found it in an old catalog from Electron Microscopy Sciences but do not have their e-mail or European adress. The 25% solution from Polysciences seems to me to be to concentrated for coating grids. Thanks in advance
Birgit
Dr. Birgit Neubohn Institut fuer Pflanzengenetik und Kulturpflanzenforschung (IPK) Corrensstr. 3 D-06466 Gatersleben-Deutschland
Does anyone have any experience with the fluorescently-tagged nanogolds? I'm starting to use one and have hit a snag; I'd like some practical input from anyone using these. The people at Nanoprobes are being very helpful, but I would like to talk to other "real-life" users, offline.
I am looking for the address of the company Technics, which produces the Hummer sputters. I know, its in Virginia. Could you mail me the whole adress (city...).
Thanks a lot!
CP Luftensteiner
I'm afraid the address I have is too old to be of any use ... if anyone on the Microscopy list can help then please reply directly to:
Vickie Allison, Group Leader, MOS 6 SEM Lab, Motorola, Inc.
--------------------------------------
Hello!
I am looking for the address of the company Technics, which produces the Hummer sputters. I know, its in Virginia. Could you mail me the whole adress (city...).
Thanks a lot!
CP Luftensteiner
I'm afraid the address I have is too old to be of any use ... if anyone on the Microscopy list can help then please reply directly to:
For those of us left working this close to Christmas, we are looking at a contaminant that appears to be mercury but are concerned about the potential hazard to both man (and woman) and machine. Are there any brave souls that have looked at mercury using a cryo system and is it relatively safe and non-destructive?? We would also like to get sample particle sizes and other potential elements present.
} Birgit Neubohn wrote: } } } } } Dear microscopists, } } } } can anybody tell me where in Germany or Europe I can buy collodion } } (celloidin, parlodion), as a powder or as a solution in amylacetate. } } I found it in an old catalog from Electron Microscopy Sciences but do not } } have their e-mail or European adress. } } The 25% solution from Polysciences seems to me to be to concentrated for } } coating grids. } } Thanks in advance } } } } Birgit } } } } Dr. Birgit Neubohn } } Institut fuer Pflanzengenetik } } und Kulturpflanzenforschung (IPK) } } Corrensstr. 3 } } D-06466 Gatersleben-Deutschland } } } } Tel.: (+49) 039482 5447 } } Fax: (+49) 039482 5139 } } e-mail: neubohn-at-ipk-gatersleben.de } } Dr. Neubohn, } } Ladd Research can supply your needs, as can many of the other supply } houses. Please contact me directly with your exact needs (i.e. } concentration). We can sell direct to you or through one of our agents } in Europe. } } Rita Arnott } International Sales } Ladd Research } 13 Dorset Lane } Williston, VT 05495 USA } tel 1-802-878-6711 } fax 1-802-878-8074 } e-mail ladres-at-worldnet.att.net
} Birgit Neubohn wrote: } } } } } Dear microscopists, } } } } can anybody tell me where in Germany or Europe I can buy collodion } } (celloidin, parlodion), as a powder or as a solution in amylacetate. } } I found it in an old catalog from Electron Microscopy Sciences but do not } } have their e-mail or European adress. } } The 25% solution from Polysciences seems to me to be to concentrated for } } coating grids. } } Thanks in advance } } } } Birgit } } } } Dr. Birgit Neubohn } } Institut fuer Pflanzengenetik } } und Kulturpflanzenforschung (IPK) } } Corrensstr. 3 } } D-06466 Gatersleben-Deutschland } } } } Tel.: (+49) 039482 5447 } } Fax: (+49) 039482 5139 } } e-mail: neubohn-at-ipk-gatersleben.de } } Dr. Neubohn, } } Ladd Research can supply your needs, as can many of the other supply } houses. Please contact me directly with your exact needs (i.e. } concentration). We can sell direct to you or through one of our agents } in Europe. } } Rita Arnott } International Sales } Ladd Research } 13 Dorset Lane } Williston, VT 05495 USA } tel 1-802-878-6711 } fax 1-802-878-8074 } e-mail ladres-at-worldnet.att.net
`Only a customer.' Ron =========================================================================== Mr. Ron Doole e-mail ron.doole-at-materials.ox.ac.uk Department of Materials, phone +44 (0) 1865 273701 University of Oxford, fax +44 (0) 1865 283333 Parks Road. Oxford. OX1 3PH. UK. ============================================================================
Lynne Garone wrote: } Hi Fellow Microscopists, } We have just installed a "windowless" Si(Li) EDS detector for our } ESEM. We would like to establish the spatial resolution of the } detector in the system as a function of our typical ESEM conditions: } 15KV, short working distance using the extended bullet, about 1 torr } pressure, gas: air, at room and cryogenic temperatures. We are curious } if fellow ESEMer's have undertaken a similar study and have some } numbers to share. Are there better conditions to optimize performance? } Any suggestions for good standards to test the spatial resolution? }
Hi Lynne,
It is possible to restore spatial resolution for EDS in the ESEM by correcting for the beam skirt effects. The methods are described in a paper which can be found at the web-address:
http://www.risoe.dk/afm/news1new.htm
best regards, Joergen.
J. B. Bilde-Soerensen Senior Research Scientist, Ph. D. Materials Research Department Risoe National Laboratory DK-4000 Roskilde Denmark
I am continuing on the subject below in seeking someones opinion on the difference betweeen the ordinary Struers Tenupol3 and a less known equipment called 550D from South Bay Technology. Are those two comparable and can 550D be used for routine work.....
Old message I am interested in your opinion about electrolyhtic thinning apparatues and their performance. I am planning to buy a jet polishing machine to be used for making thin foils from 3mm disks. The material I am interested in is High strength Aluminium alloys, steels, stainless steels.
Best wishes=20 Sten Johansson
Link=F6ping University Department of Mechanical Engineering
I just wanted to put the word out that Integrated Device Technology (IDT) has an immediate opening for a Sem Technician in our 8" Fab in Portland Oregon. The actual position is listed below along with my contact information. This is a well timed opportunity to get into a position and facility with very good growth potential. Thank you all for your time and have a great holiday season.
Dave Saiki IDT Staffing
SEM Technician
Description: Analytical SEM support for manufacturing, technology transfer, and yield enhancement. Responsible for surface and cross section sample preparation, sustaining, and imaging - Dimensional measurements (CD, film thickness) - EDX analysis. Will be working in a fully automated 8" wafer fab. New facility with a lot of growth coming.
A 2 year degree with 3+ years of experience doing Integrated Circuit construction / Failure Analysis are required for this position.
Please Respond: IDT Oregon 3131 NE Brookwood Pkwy. Hillsboro, OR 97124 503-681-6376 fax e-mail {a href="mailto:dsaiki-at-oregon.idt.com"} dsaiki-at-oregon.idt.com {/a}
We are currently looking for other options in Service Contracts for our JEOL JSM T330-A SEM. Our instrument is about 8 years old, has performed very well over this time and has been under manufacturer's Service Contract.
Because our usage has been reduced over the last couple of years, we would like to know if there are other options for maintaining and/or repairing this scope than the very expensive manufacturer's Service Contract. We are located in Philadelphia, Pennsylvania and would be interested in considering any provider who works on SEMs of this type in this region.
Any help, advice or provider recommendations would be greatly appreciated.
I beg your pardon but I failed to include my email and fax# on the preceding message.
Dave Saiki IDT Staffing
SEM Technician
Description: Analytical SEM support for manufacturing, technology transfer, and yield enhancement. Responsible for surface and cross section sample preparation, sustaining, and imaging - Dimensional measurements (CD, film thickness) - EDX analysis. Will be working in a fully automated 8" wafer fab. New facility with a lot of growth coming.
A 2 year degree with 3+ years of experience doing Integrated Circuit construction / Failure Analysis are required for this position.
Please Respond: IDT Oregon 3131 NE Brookwood Pkwy. Hillsboro, OR 97124 fax: 503-681-6376 e-mail dsaiki-at-oregon.idt.com IDT's Web Page: www.idt.com
I have owned and used both instruments as well as another instrument made by EAF Fischione that you should also consider before making you mind up. The tenepol and the Fischione model have dual polishing capability whereas the SouthBay unit only has one sided. However, the SouthBay unit can be easily converted to a chemical polishing unit for other types of materials, such as semiconductors. It can handle some pretty nasty chemicals. Another advantage of the SouthBay unit is that Bernie Kestel from Argonne National Lab has published a lot of articles on how to use this instrument for some innovative sample preparation. You can get these publications from SouthBay Technology. You should probably get a hold of them anyway since they really are definitive works on how to electropolish. The Fischione unit has a well designed sample holder that makes terminating and rinsing the sample very nice and easy. The Fischione and the SouthBay units have a reasonably sized power supply for TEM electropolishing. The Tenepol unit is huge. I modified the Tenepol unit to take the Fischione sample holder and it worked very nicely. I believe that they have modified their sample holder since I used it. If you plan to do any other type of electropolishing of large sample, the large power supply of the Tenepol unit is a plus. I used it to electroplate and electropolish parts that went into a UHV system.
Contact Dave Henriks or Shane Roberts at SouthBay Technology at "sbt-at-southbaytech.com" and Paul Fischione at EAF Fischione Instruments at "Paul.Fischione-at-internetMCI.COM"
I hope that these ramblings have helped more than they have confused you.
-Scott Walck
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
(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 am continuing on the subject below in seeking someones opinion on the difference betweeen the ordinary Struers Tenupol3 and a less known equipment called 550D from South Bay Technology. Are those two comparable and can 550D be used for routine work.....
Old message I am interested in your opinion about electrolyhtic thinning apparatues and their performance. I am planning to buy a jet polishing machine to be used for making thin foils from 3mm disks. The material I am interested in is High strength Aluminium alloys, steels, stainless steels.
Best wishes Sten Johansson
Linkvping University Department of Mechanical Engineering
Rick Felten 12/23/97 03:13 PM We need to have the stage motorized on our Hitachi S2400 SEM. The quote from Hitachi is $16,000 for X and Y w/ installation. I am sure that they have chosen a reputable sub-contractor for the job. Before we commit does anyone know of a significantly cheaper option that would still give us high quality performance?
Gerald Harrison wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello fellow microscopists, } } We are currently looking for other options in Service Contracts for } our JEOL JSM T330-A SEM. Our instrument is about 8 years old, has performed } very well over this time and has been under manufacturer's Service Contract. } } Because our usage has been reduced over the last couple of years, we } would like to know if there are other options for maintaining and/or } repairing this scope than the very expensive manufacturer's Service } Contract. We are located in Philadelphia, Pennsylvania and would be } interested in considering any provider who works on SEMs of this type in } this region. } } Any help, advice or provider recommendations would be greatly } appreciated. } } Happy year-end Holidays to all -- Gerald Harrison
Dear Gerald: Most independent service organizations are to some degree local. So it would be very helpful to know where you are located. Merry Christmas and a Happy New Year. Peter Jordan, EMSI, an independend TEM service company servicing Southern California
You could just add some solvent (probably iso amyl acetate) to your present supply. You'll have to guess the amount by adding some and watching the color of your resulting film. Start with about 10% (e.g., add 0.1 ml solvent into 1 ml commercial solution. Your film should be a light silver grey when viewed by reflected fluorescent light (like very thin silver sections). Concentration occurs through evaporation, and you can always dilute it to your specifications by experimentation. Sara
On Mon, 22 Dec 1997, Birgit Neubohn wrote:
} Date: Mon, 22 Dec 1997 14:02:02 +0100 } From: Birgit Neubohn {neubohn-at-IPK-Gatersleben.de} } To: microscopy-at-sparc5.microscopy.com } Subject: Supplier for collodion } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Dear microscopists, } } can anybody tell me where in Germany or Europe I can buy collodion } (celloidin, parlodion), as a powder or as a solution in amylacetate. } I found it in an old catalog from Electron Microscopy Sciences but do not } have their e-mail or European adress. } The 25% solution from Polysciences seems to me to be to concentrated for } coating grids. } Thanks in advance } } Birgit } } } Dr. Birgit Neubohn } Institut fuer Pflanzengenetik } und Kulturpflanzenforschung (IPK) } Corrensstr. 3 } D-06466 Gatersleben-Deutschland } } Tel.: (+49) 039482 5447 } Fax: (+49) 039482 5139 } e-mail: neubohn-at-ipk-gatersleben.de } } }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
They have a web site, www.cerious.com. Email is pcrews-at-cerious.com, phone is (704) 529-0200, fax is (704) 529-0497. You can download a shareware version from their web site to try it. The licensed version has a few more features. If you use a network, I recommend the network version. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Guys Run Rd. (packages) P.O. Box 11472 (letters) Pittsburgh, PA 15238-0472
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of S.D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
----------
Text item:
Does anyone have experience using the Polaroid Sprint Scan 45 for TEM negatives? I would appreciate any opinions on this scanner.
Thanks,
Leah L Dobbs TEM Analyst Intel Corporation
Text item: External Message Header
The following mail header is for administrative use and may be ignored unless there are problems.
There was a listing regarding an In Situ Workshop to be held in Florida. Somehow I misplaced the annoucement. If someone has the annoucement or even the phone number of the contact person, I would appreciate receiving this information. My email is 00lganion-at-BSU.edu. Thanks.
} Rick Felten } 12/15/97 12:52 PM } Does anyone know a way to print several image files in an automatic fashion } that would include the file name on the image. I am using windows 95. } } Thanks in advance.
We also use Thumbs plus. networked, multilple printers over win95/NT4 Great software!
Chris Chris Gilpin Biological Sciences Electron Microscope Unit G452 Stopford Building Oxford Road Manchester M13 9PT phone +44 161 275 5170 fax +44 161 275 5171 http://www.biomed.man.ac.uk/biology/emunit/emhome.html
Opps, I forgot to enclose my list yesterday of common problems associated with specimen collection, and tissue processing. I was hoping to get a complete list, and was hoping that other people could mention problems that they might have encountered in these areas.
Difficulties faced in the Collection of Tissue
1. Lack of the proper information on the requisitions. eg: time of fixation. Since the tissue must sit in fixative for minimum of 1 hour, if we don't know this time, then we must wait at least an hour before processing the tissue further. -name or pathology number missing, or no number on the vial, or no requisition, or a requisition with no sample. -if the requistion is not marked as a "priority", then we don't proceed to cut blue sections on the tissue, and this might result in needless delays. -illegible handwriting.
2. Insufficient sample -we often get tubes of "sample" that are apparently empty. Unless we can physically see some sample, we have trouble processing the sample because of the solution changes that we must put the sample through. If we can't see it, how do we know that we aren't "throwing the baby out with the bath water".
3. With kidney biopsy samples, the biopsy misses the glomeruli.
4. Too large of a specimen. Not only can the fixative not penetrate such a large specimen larger than a millimeter cubed, but it's extremely difficult to dehydate such tissue in the time frame that we are allowed. With incomplete dehydration, it's not possible to get the plastic to fully infiltrate the specimen, making it almost impossible to cut ultrathin, or even semithin for that matter.
Another reason why it is difficult to cut an incompletely dehyrdrated specimen block is because we cannot get complete polymerization of the plastic, so it is too soft, and does not give us enough support to cut to the very thin thicknesses required.
5. Vials with lids not tight enough.
6. Wrong specimen for the patient, or 2 specimens in the same vial, from different patients, or multiple specimens from the same patient with different tissue types in the same vial.
7. Specimen stuck to the side or lid of vial, instead of being immersed in the fixative. This might result in the specimen drying out, which of course totally destroys the ultrastructure. During the cutting of the block, it's possible to tell if the specimen has suffered this fate, because of "smoothing" problems when trying to smooth out the block surface.
8. Fresh blood or tissue sent to us directly, instead of being sent in the fixative, or sent in fixative that was poorly prepared from the referral lab. (or method of preparation was questionable.)
9. Pathologist might refer to a case by the diagnosis or tissue type, rather than the patient name or pathology number, leaving the technologist a bit baffled until they determine what case the pathologist has in mind.
10. Though it has not been a problem for us, some people might wonder what ratio of specimen to fixative should be used. In general, at least 9 parts fixative to 1 part specimen would be the largest possible ratio that one might choose.
11. Wrong container. Sperm samples have been sent in condoms, fecal samples have been sent in Cheeze Whiz jars etc, and this is not acceptable. Usually the vials supplied by our lab are adequate for most preparations, unless the sample must be centrifuged, in which case it would be preferable to sent the sample in a Falcon plastic centrifuge tube, with an adequate amount of fixative.
Problems in Polymerization
1. High humidity that we sometimes get in the summer months poses a problem for us in that the water in the air will not give us a plastic that polymerizes hard enough, consequently we have an impossible time trying to cut the specimen ultrathin. Even tbough the plastic would be hard enough for us to cut semi-thin sections, is is not hard enough to cut ultrathin.
We try to compensate through the use of dessicants and rotary vacuum pumps, to try to make our polymerization over as dry as possible, but even so, we have noticed that on the extremely humid days, the problem persists.
2. In some cases where orientation of the specimen is important, it may be difficult for us to determine the specimen orientation after fixation in Osmium Tetroxide, because the uniform black color makes it difficult to determine which side is which. In these cases reorientation after polymerization may be the only way to correct the problem.
3. Fine Needle Aspirate samples tend to give more problems in polymerization because they seem to generate more bubbles, which will often occur right at the point of interest in the specimen. We have to break all of these bubbles manually to correct the problem.
4. Use of Epon-Araldite resins, especially when combined with routine strong fixatives such as Glutaraldehyde and Osmium Tetroxide tend to destroy sensitive antigens which might have had some significance in Immunocytochemistry. This plastic also effectively renders most water soluble stains ineffective for semithin sections, essentially forcing us to adopt a monochromatic stain, rather than the preferable H+E stain which would show us contrast between the nuclei and the cytoplasm.
5. A balance must be struck between rush infiltration versus incomplete infiltration. After the sample has been polymerized though, it becomes quite useless if it has only been partially infiltrated, because at that point, it even becomes impossible to cut semi-thin sections.
Difficulties in Sectioning
1. Some tissues, because of their consistency, especially longitudinal sections of nervers, wrinkle excessively, and it is an extremely tiresome problem to deal with.
2. Plastics that have not polymerized hard enough are usually very difficult to section, and if it is possible to section, the sections break up easily of their own accord, or sometimes when under the electron beam. Usually water is to blame for this problem, especially incomplete dehydration or high humidity.
3. Plastic that is too brittle may shatter when it is being trimmed to make a useable block face. This makes it difficult to cut as well. Making plastic very quickly at high temperatures tends to result in this form of brittle plastic.
Difficulties in Staining.
1. Tissue that has stayed for too long in Glutaraldehyde my lose staining sites, and may only stain very pale, making it difficult to discern ultrastructure under the microscope.
Photography under the microscope.
1. Micrographs that are too dark result in excessive contrast. Conversely, micrographs that are too light result in prints with inadequate contrast. Micrographs that vary widely in density are difficult to print, because exposure time in the darkroom has to be changed frequently, and this results in many time wasting tests.
2. Care should be given not to shake the microscope during an exposure, or a blurred negative will result.
3. Care should be taken not to exceed the number of negatives in the electron microscope, or missing images will result.
4. With some of the older machines, one should be careful about potential camera jams, and note any unusual sounds that may occur during operation of the camera.
5. One might also be careful to note any sudden drops in vacuum after the introduction of new film. It could be that that film emulsion has not been sufficiently dried before being placed in the electron microscope.
6. Charged specimen holders or apertures can cause a slow shaking in the image that is due to charge building up and discharging. But this motion or drift can cause blurry micrographs. This can be corrected by cleaning the specimen holder in acetone, or changing the objective aperture. Increased contrast can be achieved by choosing a smaller objective aperture (of about 20 microns), and conversely, less contrast will be seen with a larger objective aperture.
7. Exact magnifications can be measured by using a calibration grid during the same session, and taking a few micrographs at the desired mag. with the calibration grid.
8. The biggest problem encountered is out of focus micrographs. This can be determined by inspection of the negative. Careful use of the wobbler focusing aid, and awareness of loss of contrast at the focused point can help reduce this problem. [some people forget to turn it off though, after using it!!] Beginners might find it useful to focus with the aid of a "hole" in the plastic, since the fresnel fringe will be clearly visible, and a large bright fresnel fringe on the inside of the hold indicates underfocus, whereas if the fringe is on the outside of the hole, it indicates overfocus.
9. The EM user should work quickly, without rushing themselves, because leaving the beam on a single area of the section burns the section. The burning might not be noticeable in the microscope, but will be readily apparent on the micrograph, and will show up as a dark circle. Excessive burning may even result in a complete penetration of the plastic by melting, resulting in total destruction of that area of the section.
10. The user should also be careful that the area of interest of the section is included on the micrograph. This is indicated on the screen by small angle marks incribed on the screen. The user should also be aware that the final image will be enlarged another 2.7 times or so in the enlarger to make the final print, and this needs to be taken into account when determining target magnification.
I will greatly appreciate if you would be able to give me a detailed protocol on how to prepare red blood cells ( I am working on fish rbc) for viewing under an SEM. Also, would it be possible to analyze for Fe using EDX.
Thank you very much. May I wish you all a Very Happy New Year.
Thanit Pewnim
%----------------------------------------------------------------------% Thanit Pewnim, Department of Chemistry, Silpakorn University Nakornpathom 73000, THAILAND } } } } } Phone +66 34 255797 Fax +66 34 255820, Internet thanit-at-kanate.su.ac.th %----------------------------------------------------------------------%
I have been tasked with coming up with something about using light microscopes without harming yourself! Not so funny, according to two of our now-retired staff who counted plankton etc. for 20+ years. They both suffered neck/shoulder/back pain which eased after retirement.
In the UK we have a specific, detailed set of Regulations concerning computer use, another unspecific set covers all workstations e.g. supermarket checkouts. A lot of the computer Regs. might be transferred to microscope set-ups.
I'd appreciate any info, anything to avoid re-inventing the wheel!
Thank you all for the replies to my query re co-visualisation of cellulose and cytoskeleton. I hope to be able to report back on the success (or otherwise) of my experiments in this area to those who requested feedback some time next year (microscope down time/holiday/trip to UK/getting hold of reagents-permitting).
In the meantime God Jul to you all,
With best wishes,
Nigel Chaffey
----------------------------------------------------- Dr Nigel Chaffey, Dept Forest Genetics & Plant Physiology, Swedish University of Agricultural Sciences, S-901 83 Ume=E5, Sweden Phone: +46-90-786-6305 =46ax: +46-90-786-5901 eMail: nigel.chaffey-at-genfys.slu.se
I have just learned that there has been a delay in the distribution of the
Call for Papers and Advanced Registration for the Microscopy & Microanalysis '98 Meeting in Atlanta, Ga July 12-16
due to a delay at the printers.
It is expected that they will be mailed during the week of Dec. 15th.
In order to avoid having to reply to repeated questions which have started to flow in my direction I have taken the liberty of posting this Email message to both the MSA Membership as well as the Microscopy Listserver Databases.
As additional information becomes available I will make sure that the M&M'98 WWW pages are updated. These WWW pages are directly accessible from the URL
Does anyone know whether the Minolta RD-175 digital camera is suitable for capturing images of biological specimens viewed with a fluorescence microscope? As I understood, this camera has an equivalent film speed sensitivity of ISO 800, which supposingly should be fast enough to capture dim fluorescent images.
Eric Cho Dept Anatomy The Chinese University of Hong Kong
can anyone of this newsgroup give me information about and subscription procedure to any related newsgroup ? In particular, I would appreciate to hear whether there is a special newsgroup discussing immuno-histology, -fluorescence, -blotting, and pathology subjects.
Thank you very much.
With kind regards,
Heinz
*********************************************************************** Dr. Heinz Fehrenbach Institute of Pathology University Clinics "Carl Gustav Carus" Technical University of Dresden
Hi Hans: Pump through the system a vinegar solution for an hour or two. If its hot its much more effective but its a bit pungent on the lungs. A bucket can serve to insert the suction and return lines a small pump is obviously required. Call me if you have any related troubles. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 77 740 370 Fax: +61 77 892 313 Great microscopy catalogue, 500 Links, MSDS, User Notes ************************ http://www.proscitech.com.au
---------- } From: H.BRINKIES {hbrinkies-at-lucy.cc.swin.edu.au} } To: microscopy-at-sparc5.microscopy.com } Subject: Cooling Water Problems } Date: Wednesday, 10 December 1997 22:01 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } -----------------------------------------------------------------------. } } Hello out there. } } I am still using an old ETEC Autoscan (SEM, vintage 1973). It is } still working well after more than 12 000 hrs of usage and usually we } get the results that we want. } } However, a calcium containing deposit has been forming } in the cooling water supply (in Cu tubes, in cooling coils around } diff.pump, in heat sinks, ect). The microscope was donated to us } several years ago but was not connected for the last 18 months } to the recirculating water system in our laboratory ( we are using } filtered tap water). The water flow has now been reduced drastically } over the last few weeks and I fear that the 'pipes' may eventually } totally block up. } } What is the best (and safe) way to reduce or remove this deposit. } Back-flashing was only partially successful. } } Any suggestion ? } } Thank You } } Hans Brinkies } SWINBURNE, University of Technology } School of Engineering and Science } Electron Microscopy Laboratory } HAWTHORN, 3122, Australia
PDC-2000 is a fine hand-held digital camera but does not have a way to adapt for the microscope. Amont other things there are internal optics that don't cooperate well with the microscope optics. It also could do some macro work with close-up lenses, but that is not optically the best imaging system.
DMC, while using the same Polaroid designed sensor chip as PDC-2000, otherwise is designed and built specifically for microscopy. Its C-mount thread (no optics) allows easy mounting on almost any microscope using standard C-mount adapters, and also can accept many "C-mount" threaded macro lenses for use on the copystand.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
On Fri, 5 Dec 1997, R-Brooks Corl wrote:
} Have you seen/tried the Polaroid DMC? It connects to the microscope } via standard C-mount (no lens on the camera, just C-mount thread), } creates a TIFF file into your computer at 1600x1200 or 800x600 pixel } resolution, and converts quickly and easily for macro work on your } copy stand by adding C-mount macro lens. List price under $6K. } Details on the Polaroid website at http:\\www.polaroid.com
How does this differ from the PDC-2000/T which, I believe, is much cheaper?
Message-Id: {199712100541.AAA28126-at-ns1.axs2000.net} To: MICROSCOPY BB {Microscopy-at-Sparc5.Microscopy.Com}
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Diane M. Smith wrote: ====================================================== Can anyone tell me about how often a diamond knife needs to be sharpened? I realize it would depend on the amount of wear it gets. Our EM dept. is only open three days a week, with cutting being done about 2-4hrs a week. Our knife was sharpened six months ago and seems to be getting dull again. Is this to be expected? ====================================================== The answer to this question is about as elusive as predicting which way the Dow Jones average will close tomorrow!
But seriously, there are the "ten commandments" for a diamond knife to enjoy a long life, the most important ones being as follows:
1] Because of the extreme sharpness of a diamond knife edge, it should not be touched even for cleaning with any solid object. This is "controversial" since some manufacturers actually "recommend" that the edges be cleaned with sticks of varying types. We ourselves believe such treatment accellerates the wearing out of a diamond knife.
2] Don't let sections or the remains of sections or other debris end up drying down onto the knife edge. Keep the knife edge "wet" until it is ready for cleaning before being put to bed for the night.
3] Use a diamond knife cleaner sold by several firms (including ours) specifically for this purpose. Some typical laboratory ultrasonic cleaners can have enough power to be damaging to a knife.
4] Wash the knife edge one last time with distilled water and then dry with some kind of "blast" such as from a clean "duster".
5] Avoid conditions of "chatter" at all times. Reduce chatter by varying the clearance angle or slowing the cutting speed. Other common causes of chatter are insufficient tightening of the boat in the microtome, an insufficiently tightened block, or an incompletely cured block.
6] Final block trimming with a razor blade can leave metal particles from the blade which are of course damaging to the knife edge. This can be minimized by using a fresh razor blade each time. Then washing the end of the freshly cut block face with distilled water, followed by a drying with a "duster" blast is the final step before the first cut with the knife. This is a final chance to wash away metal particles that could damage the knife edge.
OK, there are other considerations but these are the most important. They are independent on the knife manufacturer, the type of diamond knife, length of cutting edge, nature of the samples being cut, even the price paid.
Diamond knives in an EM lab have lifetimes that are predictable like a set of tires. It depends on what kind of road you drive on, how you do your driving, not to mention the beginning quality of the product itself. We run some samples in our own laboratory that wear out a new materials science diamond knife in a week, and we run others, e.g. soft tissue samples, that are cut with a life science diamond knife that will last, in comparision, almost forever. You can't do anything about the deck of cards you have been dealt in terms of the kinds of samples you have to cut, but once having determined that, you surely can do things, under your control that can make a big difference in terms of how long your own knife will or will not last in your own environment.
Disclaimer: SPI Supplies offers a full line of diamond knives for EM and LM . Actually we have a vested interest in having knives wear out faster rather than slower. Our favorite customers are those who mistreat their diamond knives and come back sooner for resharpenings or replacements.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Meeting announcement - Focus on Microscopy 1998 Full details below - best viewed in a monospaced font.
For a classier view see our web page created by Pal Fekete http://www.physics.usyd.edu.au/physopt/fm98
Online registration will be available on the web page soon but you can also get a form (and any further details) by email from focus98-at-emu.usyd.edu.au
Focus on Microscopy 1998 11th International Conference on 3D Image Processing in Microscopy 10th International Conference on Confocal Microscopy
April 14th-17th, 1998
University of Sydney, New South Wales, Australia
Australian Key Centre for Microscopy and Microanalysis Royal Microscopical Society (UK) Image Analysis Society of Australia
International Committee Prof. Colin Sheppard, University of Sydney Prof. G.J. Brakenhoff, University of Amsterdam Prof. Tony Wilson, University of Oxford Dr. Vyvyan Howard, University of Liverpool Dr. Andres Kriete, Liebig University, Giessen Prof. P-C. Cheng, SUNY at Buffalo Prof. Alan Boyde, University of London Dr. Guy Cox, University of Sydney Prof. S. Kawata, Osaka University
Organising Committee Prof. Colin Sheppard, University of Sydney Dr. Guy Cox, University of Sydney Ms. Carol Cogswell, University of Sydney Dr. Pal Fekete, University of Sydney Dr. Min Gu, Victoria University Dr. Allan Jones, University of Sydney Ms. Eleanor Kable, University of Sydney
Introducing Sydney
Sydney, Australia's largest city, is also one of the world's most beautiful cities, built around the spectacular natural harbour which provided the site for the first European settlement of the Australian continent. It prides itself on its cultural diversity, offering a rich mix of European, Asian and indigenous Australian experiences alongside the uniquely Australian culture which has developed in the 200 years since the First Fleet landed in Sydney Cove.
The University of Sydney is the oldest in the country, established as part of the great English 19th century tradition of liberal enlightenment but unashamedly modelled architecturally on the Oxbridge pattern. It has retained both its prestige and its central location although its site has expanded greatly over the years, now accommodating more than 20,000 students. The Australian Key Centre for Microscopy and Microanalysis (AKCMM) at the University of Sydney, which is hosting the Conference this year, is the largest centre for microscopy in the Southern Hemisphere, offering a wide range of optical and electron microscope facilities both to the University and the wider community.
April is autumn (fall) in Sydney. The weather will be mild average temperature for the month is 19 C (68 F). Sydney has both a lot of sun and a high rainfall - rain can fall in any month so bring a waterproof. The ocean will be warm and very pleasant for swimming and surfing.
Scientific Programme - 15-17 April
The scientific programme will consist of poster and spoken sessions. Posters (1m x 1m) and contributed talks (15min) are invited on any of the Conference topics:
* Advances in confocal microscopy * Applications of confocal microscopy * 3-dimensional optical imaging * 3-D techniques in electron microscopy * Other 3D imaging techniques * Novel techniques in microscopy * Near-field microscopy * Multiple-photon microscopy * Multiple-dimensional image processing * Applications of image analysis
Short Courses & Workshops - 14th April
The following half-day short courses and workshops will be offered, subject to both maximum and minimum numbers of participants. All are led by internationally recognized experts in their respective fields. The cost is $75 (Aust) per half-day course.
Morning * Multiphoton microscopy * Introductory confocal microscopy * Deconvolution of 3D images * Stereology
Afternoon * Introduction to image processing * Advanced confocal microscopy * Introduction to digital imaging * 3D image processing & visualization
Social Programme
These events are included in the cost of full and accompanying member registration. A limited number of additional tickets will be available.
* Tuesday 14th April, 6pm. Welcome reception, exhibition area. Drinks and simple snacks - an opportunity to meet old friends and to get to know fellow delegates before the start of the formal business of the conference. * Thursday 16th April, 7pm. Conference Barbecue Dinner. An informal evening on an island in one of the most beautiful parts of Sydney Harbour.
Morning coffee, a light lunch, and afternoon tea are provided each day. (For workshop registrants only on the 14th).
Abstracts
Extended abstracts, up to one A4 page in length, will be published in a conference volume issued free to all delegates. Additional copies will be available for sale. Micrographs and other illustrations (monochrome only) are welcomed but must fit within the one page.
Manuscripts will be edited for format only. To simplify the editors' task please follow these simple guidelines: * Title in upper and lower case. * Authors' names with initials first, presenting author in upper case. * Full address and affiliation of all authors. * Text in a 12pt font. * References cited by name and date, not number. * References at end - do not include titles. All authors (initials first), then year, then journal citation.
All text must be submitted electronically, either as plain text, RTF or in the format of a PC or Macintosh word processor. MS Word, Word Perfect, Wordstar, MS Works, Claris Works, Write are all on site and most other formats can be converted. Postscript files and TeX are not acceptable. Illustrations should not be included within the word processor file; they should be submitted either as separate files in any common format (not postscript or eps) or as hard copy.
Send files : * by email to focus98-at-emu.usyd.edu.au * by anonymous ftp to ftp.emu.usyd.edu.au (directory \focus98) ftp://ftp.emu.usyd.edu.au/focus98) * on a floppy disk to the conference address, below.
Abstracts must be received by 31st January 1998 if they are to appear in the published volume.
Conference Details
Conference sessions and a comprehensive manufacturers' exhibition will take place in the Wentworth Building, levels 4 & 5. Some workshops will be held in the AKCMM, Madsen Building. A footbridge across City Road provides quick access between these buildings.
A discounted early registration fee applies to all registrations received and paid by 31st January 1997. All prices given are in Australian dollars - one Australian dollar is approximately 70c US.
Early Regular Full registration $ 425 $ 475 Student registration $ 250 $ 300 Day registration $ 175 $ 175 Accompanying person $ 175 $ 175
Full registration covers conference volume, admission to all scientific sessions, welcome reception, conference dinner, morning and afternoon tea, and lunch.
Student registration includes all the above except the conference dinner.
Day registration covers conference volume, admission to scientific sessions, morning and afternoon tea, and lunch on any one day.
Accompanying person's registration includes welcome reception, two half-day tours and the conference dinner
Accommodation is available at St John's College, on the University campus, for $60 per night including breakfast (single room, shared bathroom). For those who prefer a hotel, Camperdown Travelodge is $125 per night (single occupancy) or $135 (dual occupancy) including breakfast. These are specially discounted rates - to obtain them you must book through the conference. The taxi fare from the airport to Wentworth Building, St. John's College or the Travelodge is about $10.
Focus on Microscopy '98, Australian Key Centre for Microscopy and Microanalysis, F09, University of Sydney, NSW 2006, Australia.
What are some alternatives to Absolute EtOH for drying specimens for embeddment? Absolute alcohol is hard to come by. Can someone recommend a more common substance?
I have been thinking of Acetone. My specimens will be fragile: coral tissues, sponges, worms, etc. I'd like to find something fast and good, cheap and easy.
Alan Davis --
"I consider that the golden rule requires Alan E. Davis that if I like a program I must share it adavis-at-netpci.com with other people who like it" Marianas High School AAA196, Box 10001 ---Richard Stallman Saipan, MP 96950 Northern Mariana Islands GMT+10
I would like to introduce myself as a new participant at this mailing list. I am a veterinarian (*1967) working at the Institute of Veterinary Pathology in Munich, Germany. My predominant field of research is ultrastructural enzyme- and immuno-histochemistry.
Therefore, I would like to arouse a discussion on this very topic- TEM-histochemistry, preferably post-embedding. Is there anybody out there still doing enzyme histochemistry?
Looking forward to a vivid exchange of opinions...
I would like to introduce myself as a new participant at this mailing list. I am a veterinarian (*1967) working at the Institute of Veterinary Pathology in Munich, Germany. My predominant field of research is ultrastructural enzyme- and immuno-histochemistry.
Therefore, I would like to arouse a discussion on this very topic- TEM-histochemistry, preferably post-embedding. Is there anybody out there still doing enzyme histochemistry?
Looking forward to a vivid exchange of opinions...
} } What are some alternatives to Absolute EtOH for drying specimens for } embeddment? Absolute alcohol is hard to come by. Can someone recommend a } more common substance? }
Isopropanol (or Propan-2-ol, as we are told to call it these days) in many ways behaves similarly to ethanol. It is somewhat more viscous and less volatile, but not overwhelmingly so. Moreover, it is miscible with water and should be usable for dehydrating in stages of increasing alcohol concentration.
Acteone is somewhat fiercer, might go for lipids, and when it evaporates tends to chill the specimen and pull condensation out of the air.
+------------------------------------------------------------------------+ | 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 | +------------------------------------------------------------------------+
Have not personally "looked" at metallic mercury, but have heard tales of woe from some who have tried. Mercury vapor in the sem is very bad. Will set-up a nice mercury vapor discharge lamp, disrupting everything. I have, however, examined small specimens of old amalgam without problems. You must keep the Hg from vaporizing!!!
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Seasons Greetings to all!!
For those of us left working this close to Christmas, we are looking at a contaminant that appears to be mercury but are concerned about the potential
hazard to both man (and woman) and machine. Are there any brave souls that have looked at mercury using a cryo system and is it relatively safe and non-destructive?? We would also like to get sample particle sizes and other
As a new subscriber to this group, I offer the following as a brief introduction.
My interest lie in the general area of forensic science - specifically forensic document examination. While most of my work involves optical microscopy in one form or another, I have also used SEM and CLSM to examine physical evidence in the form of "documents".
for dehydration we use either a graded series of ethanol OR acetone. This works very well, even on delicate tissues like e.g. bone marrow. You can also use propylene-glycol as the last step of dehydration, after absolute ethanol or acetone.
Yet, I'm not sure how this will work on worms. There you have the thick rigid outer shell which might impair fixation and dehydration. Have a try...
Hello, Does anyone know of a company that makes a simple micromanipulator (for making yeast tetrads) thats fitted to a Zeiss and Nikon microscopes. The original makers-Allan Benjamin Co. can not be found. Thanks.
Two vendors come to mind for micromanipulators: Leica makes excellent mechanical micromanipulators (www.leica.com), and Narishige manufactures hydraulic micromanipulators (www.narishige.co.jp). Most of these can either be placed on the microscope or put on self-supporting riser blocks that allow the needles to be brought onto the stage of the microscope.
Best regards, Bob Chiovetti ********************************* Robert (Bob) Chiovetti E. Licht Company / 1-800-865-4248 rchiovetti-at-aol.com
********************************* Leica (Wild, Leitz, Bausch&Lomb, Cambridge, AO, Reichert-Jung) / Technical Instrument Company / American Volpi / Fostek / Stocker and Yale / AEI North America / OptiQuip / Dolan-Jenner / Osram / G.E. / Philips / Ushio / Boeckler Instruments / Heidenhain / Narishige / Colorado Video / Kinetic Systems / Pryor Scientific / Compumotor / Sutter Instrument Co. / Advanced Database Systems / Cohu / Javeline Electronics / Optronics / Diagnostic Instruments, Inc. / Dage MTI / Hitachi / Panasonic / Polaroid / Kodak / Mitsubishi / Sony
My interests is in observing organic crystal growth via an optical microscope with a video/photo attachment . I would also like to study the crystallization on-line via an X-ray diffractometer. I am not after single crystals ; but a `population of crystals'.
Can anyone provide information on: -1- LINCAM - UK (manufacturer of thermomicroscopes) ; -2- in-situ crystal growth `cells' for microscopes/XRD ??
Season's Greetings! I have been asked to try to see differences in lignification in cell walls of grasses and legumes to see if it correlates with differences in expression of peroxidase. The peroxidase differences are a piece of cake to spot, but I've been having trouble with the lignin... I've tried several different protocols using phloroglucinol, but can't seem to get very intense or very consistent staining. Does anyone out there have a tried-and-true method that they are willing to share? Is there something better than phloroglucinol that I should be trying (I know it's an oldie, but seems to be standard??) thanks in advance for your help and all the best in 98!
cheers shea
Dr. S. Shea Miller Agriculture & Agri-Food Canada Eastern Cereal & Oilseed Research Centre Rm 2068, Bldg 20, CEF Ottawa, Ontario Canada K1A 0C6 Phone: (613)759-1760 Fax: (613)759-1701 e-mail: millers-at-em.agr.ca
How about a discussion on the two most important aspects of confocal microscopy as they pertain to BIOLOGICAL Raman microspectroscopy: lenses/pinholes and specimen prep. All turnkey systems I've heard about seem to be built around high-dry lenses (e.g., 100x, NA 0.9) and incorporate fixed-diameter pinholes (if they even have more than one size!). Company scientists by-and-large start to stutter and hand-wave when I mention water-immersion lenses (either "dipping" lenses or true water-as-immersion-medium types). I've seen only one BIOLOGICAL Raman paper in which the lens is mentioned; it was reported to be a Zeiss 100x, 1.2 NA water-immersion. Is anyone familiar with this lens? My sense would dictate that, if I wanted to examine hydrated biological preps of any thickness whatsoever, I would be doing myself a favor by sticking with a water-immersion lens of some type, or am I missing something here? The above comments are complicated by sample prep. To coverslip or not to coverslip, that is the question. If I would like to use the true high res water-immersion lenses, then a coverslip is required - oui, non? But doesn't this create problems with RI mismatch and thereby throw off what is already a pretty finicky measurement? Dipping lenses would seem to be the way to go here, but we need high NA and also high mag.... And what about the pinhole - I think only one turnkey device has a variable pinhole. Others have fixed pinhole(s) that, as far as I can ascertain from speaking with company scientists, have little or no relationship to the lens (?). I'd like to hear from people doing this sort of work. Rob Palmer CEB/UT
I have a theory question about Critical Point Drying that has been bothering me. I know that the specimen is placed in a "bomb", and then transition fluid replaces the dehydrating fluid in the specimen, and the temperature is raised to the critical point, which in turn raises the critical pressure in the bomb, so that the specimen is in a sense immersed in a dense vapor phase devoid of liquid air/interface, and the vapor is slowly released until the vessel is at atmospheric pressure. But with the drop in pressure, even though it is slow, below the critical pressure for that transition fluid, why doesn't this precipitate a condensation of the vapor back to a liquid????
Is the temperature slowly increased beyond the critical temperature, to correspond to a new critical temp. for the lower pressure?
For the veterans, I'm sorry to bug them with elementary questions like this, but I can't find the answer in any book.
FOR SALE: Brand New 6.2 mm Dupont diamond knife. Never used. Current retail for a knife of this length is approximately $6000+. I would like to sell the knife for $2500 or best offer. Ron Kalil
} I have a theory question about Critical Point Drying that has been } bothering me. I know that the specimen is placed in a "bomb", and then } transition fluid replaces the dehydrating fluid in the specimen, and the } temperature is raised to the critical point, which in turn raises the } critical pressure in the bomb, so that the specimen is in a sense } immersed in a dense vapor phase devoid of liquid air/interface, and the } vapor is slowly released until the vessel is at atmospheric pressure. } But with the drop in pressure, even though it is slow, below the } critical pressure for that transition fluid, why doesn't this } precipitate a condensation of the vapor back to a liquid???? } } Is the temperature slowly increased beyond the critical temperature, to } correspond to a new critical temp. for the lower pressure? } Garry,
Basically, yes. The pressure and temperature are both raised (as you say, the pressure is increased by raising the temperature). After the critical point is passed, the pressure is released *while maintaining the elevated temperature*. This means that as the pressure is lowered the CO2 is maintained in the vapor phase. Pressure *must* be released slowly both to prevent the specimen from exploding (in quotes, if you like), and to prevent the pressure drop from lowering the temperature and thus precipitating vapor condensation back to a liquid. Usually this is around 35-40 deg. C. Higher than needed to provide a safety margin.
The CO2 could be moved through the liquid-vapor transition simply by raising the temperature above its critical value, but this would mean surface tension at the liquid-vapor interface, with all the problems that implies. So the CO2 is moved through the critical point at which there is no distinction between liquid and vapor, and so there's no interface. The vapor phase is retained by keeping the temperature elevated above the critical temperture. There is no "new" critical temperature, just the same one, but CPD makes an end run around the interface problem by raising the pressure at the same time as the temperature, keeping the CO2 liquid until the critical point is reached.
My biggest problem with CPD is what exactly happens to the specimen with the increase and decrease in pressure? Supposedly nothing, if this is done slowly enough, but I don't believe it. Things may look OK, but what really happens?
Phil
} For the veterans, I'm sorry to bug them with elementary questions like } this, but I can't find the answer in any book. } } Garry
Bother us. I find it very useful to think about things I supposedly learned (mumbletymumble) years ago. Besides, someone else likely has a better explanation, and this gives me a chance to read it.
Philip Oshel PO Box 5037 Station A Champaign, IL 61825-5037 (217) 355-1143 oshel-at-ux1.cso.uiuc.edu or poshel-at-hotmail.com ***** looking for a job *****
Does anyone have experience with preparation of CoPt(20%Pt) for SEM and EBSD. Brand new material for me and aqua regia so far doesnt seem to be the right approach.Any suggestions would be appreciated. Thanks Allen Miller
%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%% %%%%%%%%% Patty Miller Stained Panes momiller-at-ccia.com
} I have a theory question about Critical Point Drying that has been } bothering me. I know that the specimen is placed in a "bomb", and then } transition fluid replaces the dehydrating fluid in the specimen, and the } temperature is raised to the critical point, which in turn raises the } critical pressure in the bomb, so that the specimen is in a sense } immersed in a dense vapor phase devoid of liquid air/interface, and the } vapor is slowly released until the vessel is at atmospheric pressure. } But with the drop in pressure, even though it is slow, below the } critical pressure for that transition fluid, why doesn't this } precipitate a condensation of the vapor back to a liquid???? } } Is the temperature slowly increased beyond the critical temperature, to } correspond to a new critical temp. for the lower pressure?
} For the veterans, I'm sorry to bug them with elementary questions like } this, but I can't find the answer in any book. } } Garry
Yes - the temperature of the whole system is kept sufficiently high so that as the pressure drops, it doesn't pass through the vapour/liquid transition again.
So the system is taken in a loop around the critical point, as illustrated:) This also highlights a minor problem that you may run into in some labs - if the ambient lab temperature is too high, the CO2 is always a vapour, so the bomb may need cooling to start with, just to get liquid CO2. This caused me some difficulties when installing a system in Jakarta at the beginning of the year!
Regards,
-- Larry Stoter 17, Rocks Park Road, Uckfield, E. Sussex, TN22 2AT, UK email: LPS-at-teknesis.demon.co.uk Phone/Fax: +44 (0)1825 767967
MicroscopyListserver Archive Email Extraction Software Version NJZ07060908