I'm doing a bit of testing right now. Hopefully you will not notice any problems, however, if you do please send an error report directly to:
Zaluzec-at-ANLEMC.MSD.ANL.GOV
I'm in the process of slowly upgrading the listserver software abit at a time to get rid of some of the annoying problems that have been creeping into the system. and the subsequent headaches for you and me. This process will likely take awhile, and I will be switching periodically between old and new links. Most of the installation was done over the holidays however, I've still got a load of reconfigurations which need to be handled "gently" to minimize any crashes. However, as Murphy has it I expect some problems. I'll keep you all informed as I make any major changes, but expect some hickups.
Subject: Time:2:17 PM OFFICE MEMO Resolved-Focusing on JEOL 1010 Date:1/3/94 Just a short note to thank everyone who responded to my request for help in rectifying our focusing problem with our the JEOL 1010. As I indicated in an earlier message, we learned a lot from the many comments and suggestions by the members of this list. It ends up that the gun alignment was changing when the shutter closed, which caused beam sensitive specimens like we use, i.e. slotted grids, to charge. Evidently, this is not a serious problem with mesh grids as are used in calibration. JEOL indicated that an early design innovation of the 1010 which was intended to blank the beam prior to exposure was later found to be less than satisfactory and abandoned this method in favor of the traditional system. Unfortunately, in our scope some remnants of this circuitry were not eliminated, thus contributing to our problem. They were able to track this problem down after considerable, and diligent effort, and corrected it by modifying the deflector lens amperage and by removing circuit elements linking the shutter and gun alignment. I am not sure what all this means, however, all or photographs since this correction have been in perfect focus. JEOL has been very cooperative and attentive to our frustrations. We are thankful that they did not take the approach of one of the members of this list who indicated that: "This is really simple! If anyone can take sharp pictures using standard specimens then the machine is working fine!" Hopefully, this information will be of use to other users of the JEOL 1010 which may encounter similar focusing problems with beam sensitive specimens.
Subject: Time:2:17 PM OFFICE MEMO Resolved-Focusing on JEOL 1010 Date:1/3/94 Just a short note to thank everyone who responded to my request for help in rectifying our focusing problem with our the JEOL 1010. As I indicated in an earlier message, we learned a lot from the many comments and suggestions by the members of this list. It ends up that the gun alignment was changing when the shutter closed, which caused beam sensitive specimens like we use, i.e. slotted grids, to charge. Evidently, this is not a serious problem with mesh grids as are used in calibration. JEOL indicated that an early design innovation of the 1010 which was intended to blank the beam prior to exposure was later found to be less than satisfactory and abandoned this method in favor of the traditional system. Unfortunately, in our scope some remnants of this circuitry were not eliminated, thus contributing to our problem. They were able to track this problem down after considerable, and diligent effort, and corrected it by modifying the deflector lens amperage and by removing circuit elements linking the shutter and gun alignment. I am not sure what all this means, however, all or photographs since this correction have been in perfect focus. JEOL has been very cooperative and attentive to our frustrations. We are thankful that they did not take the approach of one of the members of this list who indicated that: "This is really simple! If anyone can take sharp pictures using standard specimens then the machine is working fine!" Hopefully, this information will be of use to other users of the JEOL 1010 which may encounter similar focusing problems with beam sensitive specimens.
This newsgroup passed its vote by a significant margin. Barring SERIOUS controversy, it will be created in five days.
Newsgroups line:
sci.techniques.microscopy The field of microscopy.
Voting closed at 23:59:59 UTC, 2 January 1994.
After this result appears on news.announce.newgroups it will be sent to the following mailing lists (minus the individual ACK lists), with the permission of their respective maintainers:
microscopy-at-anlemc.msd.anl.gov {Microscopy Mailing List} maintained by Nestor J. Zaluzec {zaluzec-at-anlemc.msd.anl.gov}
nih-image-at-soils.umn.edu {NIH Image Mailing List} maintained by John Ladwig {jladwig-at-soils.umn.edu}
This vote was being conducted by a neutral third party. For voting questions only, contact pschleck-at-unomaha.edu. For questions about the new group, contact John F. Mansfield {John.F.Mansfield-at-umich.edu} .
(Vote-takers Note: I apologize for the lack of a 2nd CFV. It was my intention to send one out halfway through the voting period, on December 19th. However, I wasn't paying close attention to the news.announce.newgroups submission deadline before the holidays, which came up earlier this year, on December 18th. The extremely decisive results, combined with the usual lull in activity over the holidays, makes it almost impossible that this accidental oversight affected the the outcome of the vote, anyway.)
CHARTER
The main aim of sci.techniques.microscopy is to provide an open forum for the discussion of microscopy and related fields on the Internet.
PURPOSE
The purpose of sci.techniques.microscopy is to provide an open discussion forum for the microscopy community on the Internet. The newsgroups allow the rapid and timely discussion of opinions and information that would take months or years (or not at all) on conventional paper journals. It is hoped that this newsgroup will eventually be linked with the microscopy-at-anlemc.msd.anl.gov mailing list and archived at the same site. Technical suggestions as to the best way to accomplish this are welcome, and may be directed to either John F. Mansfield or Nestor J. Zaluzec via their respective E-mail addresses above.
Please note that this proposed newsgroup is intended to be an open forum for discussion of microscopy. Thus relevant topics for this newsgroup should only be limited to what the participants in this proposed newsgroup regard as microscopy.
TOPICS FOR DISCUSSION
Optical Microscopy Confocal Microscopy Scanning Probe/Force Microscopy (formerly Atomic Force Microscopy) - SPM, SFM, AFM Scanning Tunnelling Microscopy - STM Scanning Electron Microscopy - SEM Transmission Electron Microscopy - TEM High Resolution Electron Microscopy - HREM Analytical Electron Microscopy - AEM Scanning Transmission Electron Microscopy - STEM High Voltage Electron Microscopy - HVEM X-ray Energy Dispersive Spectroscopy - XEDS Electron Energy Loss Spectroscopy - EELS Electron Microprobe Analysis (EMPA) Wavelength Dispersive X-ray Spectroscopy (WDS) Diffraction Contrast Imaging Phase Contrast Imaging Selected Area Electron Diffraction - SAED or SAD Convergent Beam Electron Diffraction - CBED Image Filtering Field Ion Microscopy Electron Holography X-ray Microscopy Scanning Acoustic Microscopy Ultrasonic Imaging Specimen Preparation (Electropolishing, Ion Milling, Ultramicrotoming, etc.) 3D reconstruction Image Processing Software Data formats Databases Hardware/Equipment - specs, opinions, etc. Applications Announcements/reviews of papers/conferences. Preparation techniques. Non-ambient techniques General Discussion/opinions/questions. Positions vacant
As well as anything else that is relevant to microscopy in general.
[sci.techniques.microscopy group vote Final Vote Ack deleted to avoid overloading the mailing lists. The full results may be obtained via anonymous FTP from ftp.uu.net under /usenet/news.announce.newgroups/ sci/sci.techniques.microscopy.]
Hello, I just started working with magnetic steel specimens in the TEM and find that the beam is just "bent" everywhere. Has anyone worked with steel before and/or does anyone have any pointers they picked up along the way? Thanks..
Return-Path: {P.V.Hatton-at-sheffield.ac.uk} Received: from oxmail.ox.ac.uk by vax.ox.ac.uk (MX V3.3 VAX) with SMTP; Tue, 04 Jan 1994 14:43:36 +0000 Received: from sheffield.ac.uk by oxmail.ox.ac.uk via JANET with NIFTP (PP) id {17715-0-at-oxmail.ox.ac.uk} ; Tue, 4 Jan 1994 14:43:26 +0000 Received: from pp2.shef.ac.uk by pp.shef.ac.uk with SMTP (PP) id {04547-4-at-pp.shef.ac.uk} ; Tue, 4 Jan 1994 14:41:54 +0000 Received: from gorgon.shef.ac.uk by pp2.shef.ac.uk with SMTP (PP); Tue, 4 Jan 1994 14:33:35 +0000 Received: From LADYBOWER/GORGON_WORKQ by gorgon.shef.ac.uk via Charon-4.0A-VROOM with IPX id 100.940104143216.2368; 04 Jan 94 14:33:35 +0000 Message-ID: {MAILQUEUE-101.940104142755.448-at-UNDERBANK.shef.ac.uk} To: rms-at-vax.ox.ac.uk
I would be grateful if you could circulate the following info:
"X-Ray Microanalysis - Advances and Applications" The Spring Meeting of the X-ray Microanalysis Group will be held at the School of Clinical Dentistry, University of Sheffield on Thursday 7th April 1994. Enquiries can be directed to Dr Paul Hatton, tel. 0742 670444 ext. 3051, fax 0742 797050
Journal of Microscopy, Vol. 173, Pt. 1, January 1994, pp. 1--8.
The observation of large magnetite crystals from magnetotactic bacteria by electron and atomic force microscopy
by MARCOS FARINA,* BECHARA KACHAR,"" ULYSSES LINS,* RAYMOND BRODERICK~~ & HENRIQUE LINS DE BARROS##,
*Instituto de Biofisica Carlos Chagas Filho - CCS - Bloco G -Universidade Federal do Rio de Janeiro, 21949-900, Rio de Janeiro, Brazil. ""Laboratory of Cellular Biology, National Institute on Deafness and Other Communication Disorders, Bethesda, Maryland, U.S.A. ~~Department of Pharmacology and Experimental Therapeutics, University of Maryland at Baltimore, School of Medicine, Maryland, U.S.A. ##Museu de Astronomia e Ciencias Afins, Rio de Janeiro, Brazil
Summary Magnetite crystals inside coccoid magnetotactic bacteria found in lagoons near Rio de Janeiro city were examined by electron microscopy (EM) and atomic force microscopy (AFM). For AFM, ultrathin sections of bacteria embedded in Epon resin were etched with an ethanolic NaOH solution and observed both in the height and in the force modes. Comparative electron microscope images were useful for identifying crystalline reliefs in the etched sections. Different situations representing particular arrangements of crystal chains were observed by AFM. The majority of the bacteria examined presented unusually large magnetite crystals which remained strongly attached in linear chains even after the laboratory procedures for their isolation. This behaviour is different from all other biogenic magnetite crystals isolated so far. It is suggested that this attachment is due to the strong field between individual crystals as well as to the contact areas, which are the largest observed until now. The correct identification of a particular topography by AFM as a crystal relief may be critical when crystals are not aligned in chains; in these cases the linear dimensions and the presence of well-defined edges and faces are important features to be taken into account. Characterization of the crystal faces is important for the study of magnetotactic micro-organisms since the crystalline habits seem to be species-specific. Observation of etched sections proved to be a helpful approach for crystal relief observation, especially when small amounts of bacteria were available.
Journal of Microscopy, Vol. 173, Pt. 1, January 1994, pp. 9--25.
Development, quantitative performance and applications of a parallel electron energy-loss spectrum imaging system
by G. BOTTON* & G. L'ESPERANCE,
Centre for Characterization and Microscopy of Materials, Departement de Metallurgie et Genie des Materiaux, Ecole Polytechnique de Montreal, C.P. 6079, Succ. ``A'' Montreal, (Quebec), Canada
Summary The development of a parallel electron energy-loss spectrum imaging system is presented. The analytical performance of the imaging technique was investigated and the system applied to materials science problems. The system, which allows acquisition and storage of a parallel electron energy-loss spectrum at each pixel of an image, was developed by interfacing a multichannel analyser and a microscope to a computer workstation. In the experimental conditions used for imaging, detection limits and quantification errors were large and varied as a function of spatial resolution and the range of chemical elements of interest in the image. Applications of this imaging technique in materials science showed that quantitative chemical information is provided by the system and that the use of relative thickness maps and detailed statistical analysis of the spectrum-image allowed an unbiased interpretation of the images. As energy-loss spectra are available after processing, spectroscopic information about the analysed material can be used to provide supplementary information.
Journal of Microscopy, Vol. 173, Pt. 1, January 1994, pp. 27--38.
Reflectance in situ hybridization (RISH): detection, by confocal reflectance laser microscopy, of gold-labelled riboprobes in breast cancer cell lines and histological specimens
by G. LINARES-CRUZ,* J. P. RIGAUT,"" J. VASSY,"" T. C. DE OLIVEIRA,"" P. DE CREMOUX,* B. OLOFSSON~~ & F. CALVO* ,
*Laboratoire de Pharmacologie Experimentale, Institut de Genetique Moleculaire, Hopital Saint-Louis, Universite Paris 7, 27 rue Juliette Dodu, F-75010 Paris, France. ""Laboratoire d'Analyse d'Images en Pathologie Cellulaire, U. 263 - I.N.S.E.R.M., Universite Paris 7, 2 place Jussieu, F-75251 Paris Cedex 05, France . ~~U. 248 - I.N.S.E.R.M., Faculte de Medecine Lariboisiere Saint-Louis, 10 avenue de Verdun, F-75010 Paris, France
Summary A method for reflectance in situ hybridization (RISH) is presented. The importance of the method is demonstrated by results obtained on cytological and histological breast cancer specimens. Scattering reflectance signals from 1-nm colloidal-gold particles after RNA/RNA in situ hybridization, using digoxigenin-labelled riboprobes, were detected by confocal scanning laser microscopy. The mRNA expression of two ras-related genes, rho B and rho C, was analysed in human histological breast cancer specimens and in human breast cancer cell lines. Horizontal (x, y) and vertical (z) optical sections after three-dimensional imaging were used for visualization. A marked heterogeneity (between individual cells and between specimens) was noted for the expression of the rho B gene, both in cytological and in histological samples. On the other hand, rho C was always expressed and showed no heterogeneity. This method allows the identification of several cellular constituents in an heterogeneous tissue structure, as demonstrated by the simultaneous detection of rho B (or rho C) by reflectance and of DNA, cytokeratin and/or vimentin by fluorescence.
Journal of Microscopy, Vol. 173, Pt. 1, January 1994, pp. 39--51.
Quantitative analysis of variable-angle total internal reflection fluorescence microscopy (VA-TIRFM) of cell/substrate contacts
by J. S. BURMEISTER, G. A. TRUSKEY & W. M. REICHERT* ,
Department of Biomedical Engineering, Duke University, Durham, NC 27708, U.S.A.
Summary Variable-angle total internal reflection fluorescence microscopy (VA-TIRFM) allows controlled variation of the illumination depth with the potential of measuring both membrane/substrate separation distances and sizes of focal contacts. VA-TIRFM images are collected from well-spread bovine aortic endothelial cells (BAEC) stained with a membrane-bound carbocyanine dye. Quantitative determination of absolute membrane/substrate separation distances and individual focal contact area are attempted using a simplified model of TIRFM optics. For angles slightly greater than the critical angle of 64 degrees, both the dorsal and ventral membranes were illuminated, while images excited above 66 degrees illuminated only focal contacts. Above 74 degrees the fluorescence of focal contacts was dominated by background noise. Direct application of the simplified optical model without accounting for background intensity was unsatisfactory. However, correction for background fluorescence and nonlinear regression of the untransformed data over the working range yielded focal contact separation distances of 24 (plus or minus 13) nm. Focal contact areas estimated by TIRFM (1.3 plus or minus 0.7 square micrometres) agreed closely with areas observed by immunofluorescence staining of vinculin (1.5 plus or minus 0.3 square micrometres).
Journal of Microscopy, Vol. 173, Pt. 1, January 1994, pp. 53--66.
Semiautomated methods for cancellous bone histomorphometry using a general-purpose video image analysis system
by W. E. HUFFER,* P. RUEGG,* J.-M. ZHU"" & R. B. LEPOFF* ,
*Department of Pathology, University of Colorado Health Sciences Center, 4200 East Ninth Avenue, Denver, Colorado, U.S.A.
Summary Semiautomated methods are used to measure elongated, curved and complex branching profiles and isolated perimeter segments in monochrome video images with a general-purpose analysis system. These methods are used to make the major primary measurements of bone histomorphometry. Accuracy and reproducibility of the image acquisition, processing and measurement system is documented by measuring a semicircular standard of known dimensions. Semiautomated applications of the Ar/Le method for measuring areas and perimeters, and calculating lengths and widths of osteoid seams, lengths of mineralization labels and mineral apposition rate, wall width, indirect measurements of eroded, osteoclastic and osteoblastic perimeters without tracing, and measurement of mineralized or total cancellous bone area and perimeter gave values comparable to measurements of the same parameters by tracing or grid counting techniques with equal or better reproducibility and much greater efficiency. Intraindividual variation in measuring multiple bone biopsies was comparable to that reported with current standard methods. Major sources of variability for semiautomated methods were image magnification and selection of profile edges by thresholding, and sources of variability for manual methods are image magnification, numbers of orthogonal intercepts, tracing speed and accuracy of the algorithm used to measure traced pixels. Semiautomated methods are accurate, reproducible and rapid methods suitable for bone histomorphometry.
Journal of Microscopy, Vol. 173, Pt. 1, January 1994, pp. 67--72.
Some remarks on the accuracy of surface area estimation using the spatial grid
by K. SANDAU* & U. HAHN , Institut fur Angewandte Mathematik und Statistik, Universitat Hohenheim, D-70593 Stuttgart, Germany
Summary A set of three line grids in three orthogonal directions is called a spatial grid. This spatial grid can be used for surface area estimation by counting the number of intersection points of a surface with the grid lines. If direction and localization of the spatial grid are suitably randomized, the expectation of this number is proportional to the surface area of interest. The method was especially developed for cases where the surface to be measured is embedded in a medium, which is the usual case in microscopical applications, and where a stack of serial optical sections of the surface is available. The paper presents an improvement of an earlier version of the counting rule for intersection points. Furthermore, if the direction of sectioning is not uniform random, a bias results. This bias is calculated for a disc as a perfectly anisotropic object. A generalization of the estimator is considered by introducing a weighted mean instead of the usual arithmetic mean. The variance due to the randomized direction is investigated depending on the weights, and the minimum of this variance is derived. The relationship between the covariogram and the variance of the surface area estimated with the spatial grid is considered.
Journal of Microscopy, Vol. 173, Pt. 1, January 1994, pp. 73--78.
Volume-weighted mean particle volume estimation using different measurement methods
by E. ARTACHO-PERULA & R. ROLDAN-VILLALOBOS,
Department of Morphological Sciences, School of Medicine, University of Cordoba, Avda. Menendez Pidal s/n, 14071 Cordoba, Spain
Summary The use of the `point-sampled intercepts' method on histopathological material is evaluated for the main purpose of comparing different methods of intercept length measurements. The volume-weighted mean nuclear volume of the carcinoma of the ampulla of Vater is calculated using three methods for measuring intercept lengths: a semiautomatic image analyser, an equidistant ruler and a logarithmic ruler. Rulers of several classes and lengths are used, and the results of the volume-weighted mean nuclear volume estimations are compared. The equidistant ruler is more accurate than the logarithmic ruler. With a greater number of ruler classes and with adjustment of ruler length to the greatest nuclear intercept lengths, the systematic deviation from the true value of the volume-weighted mean nuclear volume is smaller. The volume-weighted mean nuclear volume parameter has great power to differentiate intestinal carcinoma from normal tissue.
Journal of Microscopy, Vol. 173, Pt. 1, January 1994, pp. 79--82.
A simple calibration for routine section thickness measurements using current density ratios
by Y. M. HENG,* F. P. OTTENSMEYER,"" A. L. ARSENAULT* & G. T. SIMON*,
*Electron Microscopy Facility, McMaster University Medical Centre, 1200 Main St. West, Hamilton, Ontario L8N 3Z5, Canada
Summary A method to calibrate current density ratios for the determination of specimen thickness is presented. This method uses a tilt series from a single noncrystalline specimen to create different thicknesses; these are used to generate data points to establish the relationship between specimen thickness and current density ratio. The actual specimen thickness at 08 tilt was determined to an accuracy of 5nm by a parallax method. From the calibration curves obtained, we observed that the current density ratio was sensitive to relative thickness changes on the same section of less than 1nm when a 50-micrometre objective aperture was used.
Journal of Microscopy, Vol. 173, Pt. 1, January 1994, pp. 83--86.
A simple filter system for processing small or transparent specimens
by B. CRIBB & J. ZHU ,
Centre for Microscopy and Microanalysis, University of Queensland, Brisbane, Qld 4072, Australia
Summary A novel method of attaching a fine mesh filter to the end of a disposable plastic pipette is described. Such a pipette filter can be used to exclude specimen uptake during specimen preparation procedures, particularly when processing small or transparent materials. The pipette filter-tip does not interfere with fluid exchange and is non-reactive with normal processing fluids.
THE JOURNAL OF MICROSCOPY IS AVAILABLE AT A REDUCED RATE FOR MEMBERS OF THE ROYAL MICROSCOPICAL SOCIETY, THE MICROSCOPY SOCIETY OF AMERICA AND THE INTERNATIONAL SOCIETY FOR STEREOLOGY. PLEASE CONTACT THE SECRETARY OF EACH SOCIETY FOR DETAILS.
INSTRUCTIONS FOR AUTHORS AND DISK SUBMISSION FORMS CAN BE OBTAINED FROM DR GILLIAN WILSON, THE JOURNAL OF MICROSCOPY, 37/38 ST CLEMENTS, OXFORD OX4 1AJ, UNITED KINGDOM. TELEPHONE +44 (0)865 248768, FAX +44 (0)865 791237, EMAIL RMS-at-UK.AC.OX.VAX.
I would like to thank everyone for their help in my decalcification problem. Several techniques were suggested and I am sure one of them will work for my needs. Again, many thanks!
Subject: Time:2:31 PM OFFICE MEMO TEM: Mag Specs Date:1/4/94 TEM work on steel specimens can be very difficult, because they are almost always magnetized. You may be able to reduce the magnetic inhomogenieties a bit by passing the specimens through a strong AC field, a process industrially known as 'degaussing'. I believe degaussing coils can be purchased from most electronics supply stores that deal in the television market. In use, you insert the specimen into the center of the coil and withdraw it very slowly, with the AC current flowing through the coil.
Besides doing what Robert Keller suggests, you should note the objective lens current for a non-magnetic sample and set the lens to the same value. Then you can bring the specimen to the eucentric height by noting when the image is at the minimum contrast condition.
Russell Cook Electron Microscopy Center for Materials Research Argonne Natonal Laboratory Argonne, IL
Automatic print processing is a great benefit whether or not you also have an automatic exposure system. Indeed, we had a Kodak Royalprinter for many years before we aquired our LogEtronics (now Egoltronics) EM55 automatic dodging enlarger. Even now some of our users (not many) prefer to use our old Durst enlarger although they have only two simple aids to exposure determination: a Kodak Projection Print Scale (cat. no. 1557248) which cost about $10 and an Ilford EM10 Exposure Monitor which cost about $25.
Russell Cook Electron Microscopy Center for Materials Research Argonne Natonal Laboratory Argonne, IL
Subject: Time: 2:17 PM OFFICE MEMO TEM Analysis of Diffusion Couples Date: 1/4/94 We have considered using a TEM to study diffusion couples, but preparation of thin foils is a major stumbling block. Is anyone familiar with a possible technique that could leave the diffusion structure intact at the interface between two unlike materials in a diffusion couple? Is anyone aware of reported work relating to TEM analysis of diffusion couples?
There were several articles/abstracts in the Proceedings of the XIIth International Congress for Electron Microscopy (1990 EMSA Proceedings) on this subject, mine among them. If memory serves me, there are articles in the EMSA or MSA proceedings for years prior to and after 1990. My primary sample preparation technique, and that of many otherr researchers, is to ion-mill cross-sectioned samples.
Russell Cook Electron Microscopy Center for Materials Research Argonne Natonal Laboratory Argonne, IL
Eaton Publishing invites submission of papers for peer review in consideration for publication in the new journal "Cell Vision - Journal of Analytical Morphology." The first issue of Cell Vision is scheduled for publication in May/June 1994.
Cell Vision is edited for those scientists and physicians that analyze morphology as a means of diagnosis or research. It is also intended for those who are interested in advances in immunocytochemistry, confocal microscopy, image analysis and more recent developments such as in situ polymerase chain reaction and probe scanning microsopy.
Cell Vision focuses on these novel analytical methods in morphology and their applications in biomedical research and diagnostics. Developments reported in this journal will benefit any scientist who visualizes and analyzes chemical components against the background of tissue structure.
Cell Vision will have an international circulation and will publish articles contributed by multinational authors. All articles will be rigorously peer reviewed and promise to be of very high quality. The international Editorial Board of Cell Vision, led by Dr. Jiang Gu of the Deborah Research Institute, includes many top scientists in modern morphology.
General information about Cell Vision (including instructions for authors and subscription information) is available by contacting Eaton Publishing, 154 East Central Street, Suite 201, Natick, MA 01760. You may also fax your requests to 508-655-9910 or submit electronic mail requests to internet address: fweaton-at-biotechnet.com.
Thanks for your comments. I'm headed off to look at my samples this afternoon and see how the suggestions work. For those who are wondering, I prepare my samples by cutting with a wire saw to 100microns and slowly (as possible) jet polish with perchloric until there is a hole.
We have just taken on a project which involves cutting "thin" sections of 35 um fenestrated glass beads. I am using tungsten coated glass knives broken via the "balanced-break" method with , as expected, less than desireable results. Has anyone had any experience with anything like this? Incidentaly, the beads are fixed and embedded in Spurr's resin. Any ideas for better sections would be appreciated.
Subject: Time:1:13 PM OFFICE MEMO Re} EM Atlas Date:1/5/94 {I am looking for EM atlases (atli?) for both "normal" and pathologic {tissues. {I work almost exclusively with mouse tissue, but any good atlas should {be enough of a guide. I'd appreciate any infomation or suggestions. Margaret, You did not mention the type of tissue you are looking at. If you are interested in nervous system tissue, the definitive "atlas" is, The Fine Structure of the Nervous System, by Peters, Palay and Webster. The most recent edition (3rd) is published by Oxford. Mike Mike_Schwartz-at-qm.yale.edu
} Date: Wed, 5 Jan 94 12:15:01 EST } From: meh-at-jax.org (Margaret E. Hogan) } To: microscopy-at-anlemc.msd.anl.gov } Subject: EM Atlas
} I am looking for EM atlases (atli?) for both "normal" and pathologic tissues. } I work almost exclusively with mouse tissue, but any good atlas should be } enough of a guide. I'd appreciate any infomation or suggestions. Thanks!!!! } } Peggy Hogan } The Jackson Laboratory } meh-at-aretha.jax.org
Another atlas, not the usual normal or pathological atlas, but very useful: Artifacts in Biological Electron Microscopy R.E.F. Crang and K.L. Klomparens, eds. Plenum Press, NY 1988
Organisers: Dr A Entwistle Dr C V Howard Dr H Gundlach
The Digital Imaging Light Microscopy Summer School is aimed primarily at scientists in biological and related disciplines and will comprise of a basic introduction, followed by a selection of modules.
-------------------------------------------------------------- Monday and Tuesday
Common Core - Introduction to Light Microscopy. This will consist of lectures, demonstrations and practical work on the following:
The History of microscopy; Introduction to microscopy; Limitations of the eye; Resolution, Contrast, Magnification; Refraction and lenses, geometrical optics, conjugate planes; Aperture; Illumination of the specimen in transmitted and reflected light; Lens aberrations and their correction; the choice of optical components; Diffraction and its consequences for the microscope image; Generation of contrast; Photomicrography.
Modules - The following five options are available:
Comprehensive Digital Light Microscopy - Stereology and digital imaging, digital image collection, digital image processing, digital image display and digital image storage.
Confocal Microscopy (students must bring their own specimens for study) - Fluorescent staining of samples, imaging of samples (fluorescence, reflectance and DIC techniques), visualization of 2-D and -D data sets and aspects of confocal theory.
Advanced Fluorescence Microscopy - Fluorescent staining of samples, ion ratioing methods, fluorescence decay-time measurements, fluorescence confocal microscopy.
Stereology and Digital Light Microscopy - Basic concepts of systematic random (ie unbiased) sampling in 3-D from histological material, efficient design-based stereological methods for estimating number, surface, length and volume (both manual methods and those employing image analysis systems will be covered) and methods of particle sizing in 3-D.
Techniques in Digital Light Microscopy - Introduction to stereology, introduction to confocal microscopy and introduction to digital imaging and processing.
FOR FURTHER DETAILS CONTACT THE ROYAL MICROSCOPICAL SOCIETY, 37/38 ST CLEMENTS, OXFORD OX4 1AJ, UNITED KINGDOM. TELEPHONE +44 (0)865 248768, FAX +44 (0)865 791237, EMAIL: RMS-at-UK.AC.OX.VAX.
I'm interested in the morphology of Panthera Pardus epidermis. Does anyone know if there is something unusual about the epithelial cells of this animal skin?
============================================================================= Francisco Javier Hernandez Blazquez | Universidade de Sao Paulo - Faculdade de | e-mail fjhblazq-at-fox.cce.usp.br Zootecnia e Engenharia de Alimentos | Voice: 55 195 616122 r. 265 Departamento de Ciencias Basicas/Histologia| Av. Duque de Caxias Norte, 225 CP 23 | Fax: 55 195 611689 CEP 13630-000 Pirassununga (Sao Paulo) | BRAZIL | ==============================================================================
Charles Bradley at Argonne National Laboratory has been sectioning very small bits of radioactive waste glasses embedded in resin. I believe the size of the glass pieces are about 40 microns or so. I am not sure of the resin he has been using, however. He has been using a diamond knife to get TEM sections from a Reichert-Jung Ultracut E.
Russell Cook Electron Microscopy Center for Materials Research Argonne Natonal Laboratory Argonne, IL
I have come across a structure, many times, that I have identified as a perichromatin granule. A dense body of about 30-40 nm surrounded by a transparent halo an usually associated with dense chromatin. I think this structure was first mentioned by Bernhard 20 or more years ago. Does any one know if this structure has been further characterized? Any leads on this would be greatly appreciated.
A paper by Bottke may be of interest: Bottke W (1976). Chromosome-associated paracrystalline nuclear inclusions in the spermatocytes of a pulmonate snail, Planorbarius corneus L. Chromosoma (Berl.) 55: 273-287.
On Thu, 6 Jan 1994, Greg Erdos ICBR EM Core Lab University of Florida wrote:
} Cell Biologists: } } I have come across a structure, many times, that I have identified } as a perichromatin granule. A dense body of about 30-40 nm surrounded by a } transparent halo an usually associated with dense chromatin. I think this } structure was first mentioned by Bernhard 20 or more years ago. Does any } one know if this structure has been further characterized? } Any leads on this would be greatly appreciated. } } ***************************** } * Greg Erdos * } * Director, ICBR EMCL * } * University of Florida * } * Gainesville, FL 32611 * } * gwerdos-at-gnv.ifas.ufl.edu * } * 904-392-1295 * } ***************************** }
With encouragement from a number of you folks out there, I tried sectioning these thing with an old diamond knive. Much to my amazement, it worked beautifully. As a biologist, I never would have thought that this was possible. We have good sections with minimal obvious damage to the knife. We have saved a great deal of time and many headaches. Thanks to all who responded and HOORAH for the system.
We are looking for a used but recent vintage electron microprobe to replace a very old ARL system. Preferences are for a 4-channel system with EDS, something like a Cameca SX-50. We are aware of Don Lesher's operation in northern Ohio and have talked with him about acquiring a rebuilt ARL-SEMQ. This may be our eventual route, but if there is a good recent vintage machine sitting out there somewhere we would be very interested in knowing about it. Thanks. Warren D. Huff Dept. of Geology University of Cincinnati Cincinnati, OH 45221-0013 phone (513) 556-3731 fax (513) 556-6931 e-mail huff-at-ucbeh.san.uc.edu
Due to the continuing recession in California and especially the hardships the UC system is undergoing we are forced to begin a recharge for our microscopy services. I would very much like to know what current recharges are for biological TEM use and prep services. If anyone has a freeze fracture device (we have a Balzers BAF 400T) what are the going rates? How about for SEM prep?
Thank you in advance. Rick A. Harris Electron Microscopy Evolution and Ecology Univ. of Calif. Davis, CA 916 752 2914
None Re: Recent request for help on volume calculations -
"Our laboratory has expressed interest in calculating volumes on microscopic data and I am looking for software that can perform the following tasks:
2: 3-D reconstruction (including EM image alignment) 5: Wire-frame generation and simple volume rendering 3: Volume calculation 1: Run on a Mac or PC 4: File I/O from a Mac or PC format (preferrably using MacDraw objects of known dimensions)
The non-proprietary file format support is neccessary to allow cross-platform data manipulation. My U*NIX access is limited, so ability to un on a PC is important.
I keep reading how people did 3-D reconstruction, but their programs are hard to find. Any referrals to sources of this software is appreciated. It WOULD be neat if it was available in C by anonymous FTP, but perhaps this is too narrow a requirement. Any leads would help."
Reply - There are simple methods for estimating volume densities that do not require complicated computer systems or sophisticated software. If your aims are to estimate the volumes of subcellular structures and you can directly measure at least one reference volume then you might like to try cross latice overlays as described by Weibel as far back as 1979 (Stereological methods vol 1. practical methods for biological morphometry Academic Press NY). More recent reviews can be found in APMIS (Gundersen et al 1988 vol 96 379-394), J. Microsc (Cruz-Orive 1982 vol 125 89-102) Am. J. Physiol (Cruz-Orive & Weibel 1990) vol 258 148-156) and TICB (Luquoc 1993 some time this fall). Another source is a recently published book where the final chapter covers all these methods (Griffiths 1993 Fine Structure Immunocytochemistry, Springer Verlag, Heidelberg). Best of all is to take a course on stereology (there is one in Banff, Canada in May 1994 and one here at Yale in August 1994). If you still want to do 3-D reconstructions, which will only give you information on the structures you reconstruct, then the best software I have seen up to now are the VoxelView programs. We run them on a Silicon Graphics workstation which never seems to have enough memory. These programs allow you to reconstruct sections, rotate and manipulate the images as well as measure the parameters of the reconstructed structures.
Immunocytochemistry and Cryosections Practical Course 22 - 27 August 1994. An intensive practical course mixed with theoretical sessions where you can learn how to produce cryosections as well as immunolabeling, colloidal gold production and much more.
Additional we are offerring a three day practical workshop on Stereological methods. This will be on 18 -20 August 1994, prior to the cryosectioning course.
A team of instructors headed by Hans Gundersen will take you through the theoretical and practical details of modern stereological methods. These will include volume and surface densities, the fractionator, the nucleator, the disector and much much more. Address for further details on both courses;
Paul Webster, Department of Cell Biology, Yale School of Medicine, 333 Cedar Street, New Haven, CT 06510.
Re} Stereology Re: Recent request for help on volume calculations -
"Our laboratory has expressed interest in calculating volumes on microscopic data and I am looking for software that can perform the following tasks:
2: 3-D reconstruction (including EM image alignment) 5: Wire-frame generation and simple volume rendering 3: Volume calculation 1: Run on a Mac or PC 4: File I/O from a Mac or PC format (preferrably using MacDraw objects of known dimensions)
The non-proprietary file format support is neccessary to allow cross-platform data manipulation. My U*NIX access is limited, so ability to un on a PC is important.
I keep reading how people did 3-D reconstruction, but their programs are hard to find. Any referrals to sources of this software is appreciated. It WOULD be neat if it was available in C by anonymous FTP, but perhaps this is too narrow a requirement. Any leads would help."
Reply - There are simple methods for estimating volume densities that do not require complicated computer systems or sophisticated software. If your aims are to estimate the volumes of subcellular structures and you can directly measure at least one reference volume then you might like to try cross latice overlays as described by Weibel as far back as 1979 (Stereological methods vol 1. practical methods for biological morphometry Academic Press NY). More recent reviews can be found in APMIS (Gundersen et al 1988 vol 96 379-394), J. Microsc (Cruz-Orive 1982 vol 125 89-102) Am. J. Physiol (Cruz-Orive & Weibel 1990) vol 258 148-156) and TICB (Luquoc 1993 some time this fall). Another source is a recently published book where the final chapter covers all these methods (Griffiths 1993 Fine Structure Immunocytochemistry, Springer Verlag, Heidelberg). Best of all is to take a course on stereology (there is one in Banff, Canada in May 1994 and one here at Yale in August 1994). If you still want to do 3-D reconstructions, which will only give you information on the structures you reconstruct, then the best software I have seen up to now are the VoxelView programs. We run them on a Silicon Graphic workstation which never seems to have enough memory. These programs allow you to reconstruct sections, rotate and manipulate the images as well as measure the parameters of the reconstructed structures.
There is a tutorial on quantitative morphometry written by Dr. Robert Bolender, Dept. Biological Structure at Univ. Washington. It also provides templates for point and intersect counts and for QM computations. It is available from the Health Sciences Center for Educational Resources, University of Washington SB-56, Seattle, Washington, 98195 (206) 685-1156.
Dr. Bolender teaches a course on QM in even years.
Glen MacDonald Hearing Development Laboratories RL-30 University of Washington Seattle, WA 98195 (206)543-8360 glenmac-at-u.washington.edu
Subscribers who might find it difficult to attend the stereology courses in Banff or Yale should note that the RMS will be running a Digital Imaging and Stereology course at Liverpool University, UK, in July 1994, coordinated by the current President of the International Society for Stereology, Dr Vyvyan Howard. Contact RMS-at-UK.AC.OX.VAX with your full name and address to obtain details!
From Paul Sheppard, Tree-Ring Lab, Univ. of Arizona
To microscopy forum members:
I am having slight, but definite, difficulty in attaining repeatable focus of my binocular common main objective microscope. In my attempt to apply image- analysis techniques to tree-ring science, my focus problem has led to systemat- ic differences in values as measured by different technicians. I am amazed at how little the focus differences must be before we see differences in our data. Can anyone suggest ways to ensure repeatable focus?
I have inquired into adding on autofocus hard- and software, but my first estimate on that was $7,000, which is prohibitive at this time. I have also heard about a dual-light focus aid, where two spot beams are projected onto the subject to intersect exactly at the focal distance of the objective lens; if the subject is above or below that distance, then the spot will be elongated or even split into two. Has anyone tried this? I have also tried focussing at high magnification and then working at my usual lower magnification. This re- quires parfocal optics, which I have, but this process is cumbersome and other- wise prone to error.
Thanks in advance for any and all suggestions,
Paul Sheppard Laboratory of Tree-Ring Research University of Arizona GRAD12-at-CCIT.ARIZONA.EDU
My work is in the materials sciences where for reflected light microscopy specien preparation usually begins with a grinding and polishing sequence in order to obtain an optically smooth surface. I am now faced with a material I do not wish to grind/polish, I passed by a reference that suggested that if the specimen surface is not optically smooth I can use glass coverslips and an index matching fluid - this sounds vaguely familar from highschool biology. Is there anyone out there with experience in Light Microscopy who could point toward the correct products and describe for me some of the considerations involved.
} Rick Harris asked about charges for EM facilities..... } --------------------------------------------------- } } Sandy Silvers of the South Eastern EM Society has a fairly } comprehensive database on EM facilities in the US. You } can contact her at
** some stuff deleted **
I just got off the phone with Sandy Silvers and told her that, since she doesn't currently have Internet access, I'd pass this information along. She has moved from Florida. Her current address and phone number are:
Sandra Silvers EM Complex USDA, ARS, RRC PO Box 5677 Athens GA 30613-6199 (706) 546-3471
Her database has listings, which include contact people, for about 200 facilities. Information about usage charges is included for most of them. She is looking for more input and will send a questionaire on request. Printed copies of the database are available for $8 (US). She does all this personally now, so needs to recoup printing costs.
She's intrigued with the possibility of having the information available more widely, perhaps through FTP.
John chandler-at-lamar.ColoState.EDU Fort Collins, CO
Subject: Time:4:14 PM OFFICE MEMO LM???-Reply Date:1/11/94 I do not know the nature of your specimens, but I seriously dobut that you would gain much useful information by using a cover slide and oil to overcome the usual polishing process. Polishing is usually necessary because optical microscopes have such limited depth of field (Typically about 10 microns for a 10X objective, 1 micron for a 40X obj, and 0.1 or 0.2 microns for a 100X obj) that they are not at all suited for looking at rough surfaces. Placing a layer of oil and a cover glass over a rough surface will not solve this problem. Furthermore, the objective lenses of metallographic microscopes are not designed to work through cover glasses - you'll probably need to go to the biology or mineralogy dept to find a microscope that is. I would suggest that you try examining the specimen with a stereo binocular microscope as a start. A good instrument of this kind should give magnifications up to about 75X. If that is not sufficient, then the next step would be to try SEM - although contrast may be a problem, depending on the nature of the specimens. microscope will not work with a cover glass,
The previously posted information about Sandy Silvers address was incorrect. She has since moved, her new address is apparently.... Thanks to L. Melsen for the correction....
Rick Harris asked about charges for EM facilities..... ---------------------------------------------------
Sandy Silvers of the South Eastern EM Society has a fairly comprehensive database on EM facilities in the US. You can contact her at
SANDY H. SILVERS EM COMPLEX RUSSELL RESEARCH CENTERS USDA, BOX 5677 ATHENS, GA 30613-6199 (706) 546 3471 FAX (706) 546 3452
Subject: Time:5:15 PM OFFICE MEMO Stereographic Calcs Date:1/11/94 We have obtained optical goniometric measurements of angles between the faces of about 75 macro crystals. These angles (commonly called phi and rho) were measured relative to the axes of the optical goniometer. Now, we want to convert them to angles that are relative to principal planes of the crystals. Does anyone have , or know of, a readily available computer program for doing this?
We are working on a project that requires us to shadow myosin with Pt and C. We are trying to see the myosin heads and another conjugated molecule. We are using our Balzers 400 to shadow at 15 to 25 degrees for the Pt and then at 90 degrees for the C. We can find the molecules but they are extremely faint. We have used anywhere from 5 to 15 angstroms of Pt. Best results were with higher angles and thinner coats. Then we increase exposure in the TEM to 3 seconds to bump the contrast and shift the s/n ratio. Has anyone a suggestion for making the myosin more visible? The micrographs have little contrast between the molecule and the substrate.
Rick A. Harris Electron Microscopy Evolution and Ecology Univ. of Calif. Davis, CA
Excerpts from mail: 11-Jan-94 LM - ??? by Peter Joyce-at-utxvms.cc.ut } My work is in the materials sciences where for reflected light microscopy } specien preparation usually begins with a grinding and polishing sequence } in order to obtain an optically smooth surface. I am now faced with a } material I do not wish to grind/polish, I passed by a reference that } suggested that if the specimen surface is not optically smooth I can use } glass coverslips and an index matching fluid - this sounds vaguely familar } If I understand your question, I don't think index matching will help. Reflections occur where refractive index changes, if you match the refractive index of your specimen you not see much reflection. I am not a materials scientist, but biologists use reflected light microscopy to generate contrast by interference of the first surface reflection (usually the coverslip/medium interface) with the second surface reflection (usually the medium/specimen interface) in order to see regions where the specimen makes close contact with the coverslip. If your application is based at all on similar effects, index matching will at least attenuate one of the reflections.
As part of a research project, I am looking for -any and all- references that pertain to the staining of mineral species for their characteriza- tion and to make certain minerals more distinguishable under the optical (polarizing) microscope.
I greatly appreciate all info and help in this endeaver.
Thanks, Rob
X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X X Rob Tayloe X MSM Spelunkers Club X X Metallographic Lab. X Missouri Speleological Survey /-v-\ \-v-/ X X Rolla Research Center X Bat Conservation International X X U.S. Bureau of Mines X Missouri Cave & Karst Conservancy \-v-/ X X tayloe-at-rorc.usbm.gov X National Speleological Society #32993 /-v-\ X X (314) 364-3169 x247 X American Cave Conservation Association X X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X
I am wondering if anyone out there has tried incorporating the new Mac AV computers with their video systems? I've got the money for the Mac, but I need to know how the software is for image analysis, and how the final product actually looks.
If anyone has done an EM video with the Mac I would like to see it.
Thanks for the help...Go Giants...long live the VW!
John L. Grazul BBR EM Facility Rutgers University Box 1059 Piscataway, NJ 08854
On the NIH-Image mailing list, many, many postings have gone through with the general concensus being that the AV input on the Macs was not intended for scientific quality. There are a number of high-quality frame grabbers for 640 x 480 pixel pictures (e.g., Scion LG-3 at 301-695-7870, Data Translation QuickCapture at 508-481-3700, Perceptics at 615-966-9200, others) as well as some higher-end systems for cooled CCDs, etc. There are also some slow-scan TEM interfaces for the Mac (Gatan, 4pi Analysis at 919-489-1757 for STEM, others). There has been considerably less discussion about the video output. If this is the issue, someone else should comment.
As for image analysis software, there are a number of packages ranging from classical microscopic image analysis to part inspection on assembly lines. My choices are NIH Image (free from NIH via anonymous FTP to zippy.nimh.nih.gov) for workhorse viewing and automation as well as "simple" particle analysis and PrismView (marketed by Signal Analytics 703-281-3277) for high-end work. NIH Image is quick to learn, readily extensible and is extremely popular with its infinite return-on-investment! There are other systems as well.
These vendor lists are NOT exhaustive. If you can download NIH Image, appendices B,C, and D are a little more exhaustive for Mac imaging vendors.
Bill ========================== Bill Heeschen / Analytical Sciences - Materials Characterization 1897-D Building / The Dow Chemical Company Midland, MI 48667 U.S.A. phone: (517)636-4005 fax: (517)636-5453 Email: waheeschen-at-dow.com ==========================
} } As for image analysis software, there are a number of packages ranging from } classical microscopic image analysis to part inspection on assembly lines. My } choices are NIH Image (free from NIH via anonymous FTP to zippy.nimh.nih.gov) } for workhorse viewing and automation as well as "simple" particle analysis and } PrismView (marketed by Signal Analytics 703-281-3277) for high-end work. NIH } Image is quick to learn, readily extensible and is extremely popular with its } infinite return-on-investment! There are other systems as well. } } These vendor lists are NOT exhaustive. If you can download NIH Image, } appendices B,C, and D are a little more exhaustive for Mac imaging vendors. } } Bill
Another note regarding easy info on NIH Image, subscribe to the mailing list located on nih-image-at-soils.umn.edu. They're alll the time fielding most any question you can think of, including questions regrading PrismView. If it's too much to subscribe, post your question anyway and have replies sent directly to you. Good luck.
I was wondering if there was a software package similar to NIH-Image that will run on the IBM or Silicon Graphics workstation that one can get through ftp.
I need a small sample of Ti203 for some microscopy work I'm doing. It's only a really small amount needed so If someone has some that they can give me I would be most appreciative, failing that does anyone know of a supplier?
Thanks David
-- ---------------------------------------------------------------------- David Bell |E-mail: dcb-at-electron.ph.unimelb.edu.au School of Physics |Phone : +61 3 344 5451 The University of Melbourne |Fax : +61 3 344 4783 Parkville, Victoria, AUST, 3052 |
Here is a supplier for Ti2O3, otherwise known as titanium (III) oxide: AESAR/Johnson Matthey 30 Bond Street P.O. Box 8247 Ward Hill, MA 01835-0747 (800) 343-1990 (508) 521-6300. You should be able to get 50 grams for about $50.
J. Ester asked } I was wondering if there was a software package similar to NIH-Image } that will run on the IBM or Silicon Graphics workstation that one can } get through ftp
There is no equivalent for NIH Image that will run on the PC. The closest thing (and it's not close) is NCSA Image fro the National Center for Supercomputing Applications at the Univ. of Ill. It is available via FTP. Their address is "ftp.ncsa.uiuc.edu". NIH Image as I understand from Wayne Rasband (the author at NIH) will be ported to the PowerPC when he gets a unit. So those of you who are tied to PC will be able to run as long as you run the machine in it's Mac Compatible Mode instead of the PC Compatible Mode.
Department of Anatomy and Neurobiology Colorado State University Fort Collins, CO
July 11-15, 1994
The CSU Electron Microscopy Center in the Department of Anatomy and Neurobiology offers an intensive course in freeze-fracture and freeze-etch techniques for research scientists and senior technicians.
Basic and advanced freeze-fracture and freeze-etch techniques will be taught in five days of concentrated lectures and laboratory sessions (12-15 hr/day). Advanced techniques will include sequential confocal mapping/freeze-fracture examination of identified cells in tissue slices. This course will prepare research scientists and laboratory technicians to use freeze-fracture techniques in cell biology research. No previous freeze-fracture experience is necessary, but the individual must be proficient in transmission electron microscopy.
Participants will prepare high-resolution replicas of their own specimens, become proficient at interpreting the resulting images, and learn to prepare and label their own stereoscopic micrographs. Faculty and participants will discuss the physical and chemical bases for interpreting freeze-etch replicas, the major sources of specimen preparation artifacts, and the unique advantages and limitations of freeze-fracture and freeze-etch techniques. Methods for reducing specimen contamination during the fracturing and replication processes, as well as techniques for identifying, fracturing, and mapping individual cells in tissue slices, will be described.
This course is organized by Drs. John Rash, John Walrond, Robert Lee and John Chandler in collaboration with major instrument manufacturers.
Freeze-fracture/freeze-etch equipment used in the course includes: Balzers BAF-301 and BAF- 400, JEOL JFD-9010-CR and additional rapid freeze and freeze-etch devices supplied by RMC, Inc. and Bal-Tec, Inc. Molecular Dynamics multi-probe 2001 confocal laser scanning microscope will be used.
Registration fee of $1250 includes textbook, laboratory instruction manual, all necessary supplies and implements, noon meals, refreshments, EM negatives and prints, and unlimited use of equipment during the course.
Information concerning this course, hotel accommodations and travel arrangements may be obtained from:
Eileen Diepenbrock Colorado State University Department of Anatomy and Neurobiology Fort Collins, CO 80523 (303) 491-5847 ediepenbrock-at-vines.colostate.edu
Registration closes May 15, 1994
Course registration will be for a minimum of 10 and maximum of 12 participants.
John chandler-at-lamar.ColoState.EDU Fort Collins, CO
I've had multiple requests lately for permission to post resume's and job "hunting notices" on this listserver. As we set forth early in the setup of this discussion/information forum that type of posting is not appropriate here. However, I realize that with the current budget situations across the country & world there will be a significant number of highly talented individuals looking for positions in the near term. Allow me then to remind all of you that should you have acess to information concerning research/teaching positions this type of BULLETIN/ANNOUNCEMENT is allowed on the Mailserver and I would encourage you to post it. If you are not sure about any posting feel free to Email it directly to me and I will review it's appropriateness.
Individuals interested in posting (SHORT) electronic resume's are welcome to access the ANLEMC/MSA electronic bulletin board. It can be reached via INTERNET/TELNET link at the address: ANLEMC.MSD.ANL.GOV, login with the username EMCBBS and password EMCBBS then follow directions on the screen. Please note that there is only a single connection from INTERNET to this BBS line and that the line may be in use. Pay attention to any messages on your terminal when you login this will clue you in to what is happening....
I received a copy of your response to John L. Grazul regarding digital TEM. We are using a system consisting of a Gatan wide angle video camera (model 673mkIII ), a Dage video processor (DSP200), Quick Capture board and a MacII running NIH-Image. The combination yields 640x480 images directly from the microscope. With the frame processor we can average frames (2,4,8,16,32) and adjust the gain and offset of the video signal so the Mac receives a full range signal. I have written some macros for Image to permit rapid specimen ID, magnification setting and storage. Folders of images are then sent over the network to the pathologist's desk.
We have found that the images are sufficient for diagnostic purposes and have begun to shift to electronic imaging.
The images are NOT the quality of film, but the cost for the system is low (NIH Image is free and VERY good). My concern was (and is) that with a slow scan system the increased cost doesn't really get you that much more in terms of resolution (maybe twice the resolution for lots more money). The microscope has such excess magnification that it is much cheaper to acquire multiple images at higher magnification (and hence greater resolution) than it is to purchase a slow scan camera and the associated software drivers. My thought is that if you want film-like resolution you should take micrographs and either print them or digitize the negative with a flatbed or drum scanner for image analysis. If all you want to do is some image analysis, it is entirely possible that a 640x480 image will be sufficient.
Feel free to forward this to Grazul and others on whatever mail list or news group you are on.
I would be interested to learn how you are using your system.
Charles Daghlian Rippel E. M. Facility Dartmouth College Hanover, NH 03755 603-650-1337
The list administrator is John Ladwig (jladwig-at-soils.umn.edu), but don't bug him until you've tried the above instructions! 8-)
As many of you list participants are aware, there is movement afoot to set up a NewsGroup which will be a superset of both the nih-image and microscopy lists. As this develops, there will be postings to both lists describing how/when this happens.
Bill ========================== Bill Heeschen / Analytical Sciences - Materials Characterization 1897-D Building / The Dow Chemical Company Midland, MI 48667 U.S.A. phone: (517)636-4005 fax: (517)636-5453 Email: waheeschen-at-dow.com ==========================
* SC3 Ion Projection Lithography Instructor: John C. Wolfe, Univ. of Houston
Resists -------
* SC4 Introduction to Microlithography, Resist Materials and Processing Instructors: Larry F. Thompson, AT&T Bell Labs.; Murrae J. Bowden, Bell Communications Research, Inc.; C. G. Willson, Univ. of Texas/Austin
* SC5 Optical Lithography Modeling Instructors: Chris A. Mack, FINLE Technologies, Inc.; Andrew R. Neureuther, Univ. of California/Berkeley
* SC6 Resist Thickness, Bake, Exposure, and Development Control Instructor: W. Tom Batchelder, Semiconductor Systems, Inc.
* SC7 Resists for Deep-UV Lithography Instructor: C. Grant Willson, Univ. of Texas/Austin
* SC8 Diazonaphthoquinone-Based Resists Instructor: Ralph Dammel, Hoechst Celanese Corp.
Metrology & Process Control ---------------------------
* SC9 IC Critical Dimension Measurement System Instructors: Sadri Khalessi, Metrologix, Inc.; Kevin M. Monahan, Metrologix, Inc.
* SC10 Fundamentals and Pitfalls of Submicrometer Dimensional Metrology Instructor: Terrence E. Zavecz, TEA Systems Corp.
* SC11 The Physics and Simulation of Metrology Instruments Instructor: Mark P. Davidson, Spectel Co.
* SC14 The Use of the Low-Voltage SEM in IC Fabrication Instructor: Michael G. Rosenfield, IBM Thomas J. Watson Research Ctr.
Microlithography ----------------
* SC15 Advanced Topics in Optical Lithography Instructor: Chris A. Mack, FINLE Technologies, Inc.
* SC16 Fundamentals of Cameras and Projectors Instructor: Warren J. Smith, Kaiser Electro-Optics, Inc.
* SC17 Introduction to Optical Lithographic Tools Instructor: Timothy A. Brunner, IBM Thomas J. Watson Research Ctr.
* SC18 Plasma Etching Technology Instructor: Daniel L. Flamm, Univ. of California/Berkeley
* SC19 The Exposure-Defocus Tree and Its Uses in Understanding and Extending Optical Lithography Instructor: Burn J. Lin, Linnovation, Inc.
* SC20 Optimization Methods for Microlithographic Materials and Processes Instructor: Daniel J. Herr, Semiconductor Research Corp.
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Don Grimes of Microscopy Today has sent the enclosed message along to me for approval for posting on the Microscopy Mailserver. I do not have a problem with it as it is a general announcement meant as a service to Microscopists who will be looking for jobs.
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To: Membership
Thanks to many for your interest and comments regarding our newsletter. Several have inquired over the possibility of our assisting in their search for new employment. I have previously not done this as it does not quite meet my objective as "material of interest to a reasonable number of microscopists". However I would like to try to help. If you seek new employment and would like to provide me a summary of either "what" you are or "what" you are looking for, I will publish the summary in my next issue. I only ask that you keep the summary under around 60 words and I MUST have it no later than this coming Friday (21 Jan) to make the issue. And if you wish to keep your name "quiet", we can do the Box # thing - where a number rather than your name is listed, and I will forward any interests direct to you. I expect that my newsletter is received in 99+% of the microscopy labs in the U.S. but can not guarantee that it goes to the right person.
Due to the number of replies to the attached message here is more info. The QM2000 tutorial runs on DOS and Windows. Its development platform is due out for the Mac and Unix, so there may be ports to those operating systems. Dr. Bolender has released a quantitative morphometry dataabase for the nervous system, developed with Sybase, running on a Sparc.
A summer quarter course in quantitative morphometry will probably be taught here this summer.
For more information: Dr. Robert Bolender Dept. Biological Structure SM-20 University of Washington Seattle, WA 98195 rpb-at-u.washington.edu
******old message There is a tutorial on quantitative morphometry written by Dr. Robert Bolender, Dept. Biological Structure at Univ. Washington. It also provides templates for point and intersect counts and for QM computations. It is available from the Health Sciences Center for Educational Resources, University of Washington SB-56, Seattle, Washington, 98195 (206) 685-1156.
Dr. Bolender teaches a course on QM in even years.
Glen MacDonald Hearing Development Laboratories RL-30 University of Washington Seattle, WA 98195 (206)543-8360 glenmac-at-u.washington.edu
I will am about to start as a postdoctoral scientist working with Prof. Kirz at the State University of New York at Stony Brook/Brookhaven National Laboratory in the field of x-ray microscopy. I am very interested in your EM-course on Immunocytochemistry and Cryosectioning. Please let me know what I need in order to apply. Sincerely, Joerg Maser
The Department of Materials Science and Engineering at Lehigh University has an opening for a senior (associate/full) professor with expertise in electron microscopy and/or microanalysis (SEM, TEM, AEM, etc.). The successful candidate must have a proven record of research in the application of microscopy and/or microanalysis to the solution of materials problems and be able to conduct independent and cooperative research in MS&E. Ability to teach undergraduate and graduate courses in microscopy and materials is essential. Experience in running a microscopy facility will be an advantage. Position available September 1, 1994.
Curriculum vitae and the names of three references should be sent by May 15, 1994 to: Professor David B. Williams Chairman, Search Committee Dept. of Materials Science and Eng. Lehigh University 5 East Packer Avenue Bethlehem, PA 18015-3195
Lehigh University is an equal opportunnity employer and welcomes applications from all qualified candidates.
Thanks to B.Miner for forwarding the useful comments from P.Catinella and B.Edwards on RIE etching on microelectronic devices.
One other question that I would like to direct to them or other interested parties would be the requirement for installing a "scrubber" on the exhaust fumes from the RIE. We are working up a estimate for the possible total cost of purchasing/operating/maintaining a plasma etcher in our laborato- ry. The requirement for a "scrubber" on the exhaust has been mentioned as a possibile environmental requirement. I have spoken to some manufactures and they don't use scubbers due to low flow rates in their machines and small quantities of gas involved.
Thanks again.
Richard Sartore US Army Research LAboratory AMSRL-EP-RA Fort Monmouth, NJ 07703 RSARTORE-at-MONMOUTH-ETDL1.ARMY.MIL
To Membership, International (or local) readers who wish to receive a no cost copy of our newsletter "Microscopy Today" and are having trouble contacting me, as I am on Compuserve, are encouraged to do so by FAX. My number is (608)836-1969. And the several who have requested the newsletter but did not supply their addresses are encouraged to resent their requests.
We have a Nanoscope III here, and would like to study the images off-line on another pc. Does anyone know good image analysing & processing programs for the PC? Basic filtering and measurement functions (cross sections etc) are a must.
I've heard names like Global Lab Image, Mocha, Image Pro Plus. Any experiences on these? Where to take contact: companies, addresses, fax numbers?
Thanks in advance,
Markus Levlin
-- Markus Levlin Laboratory of Physics tel +358 0 451 3144 markus.levlin-at-hut.fi Helsinki University of Technology fax +358 0 451 3116 Otakaari 1 M, 02150 Espoo, Finland
Thanks to B.Miner for forwarding the useful comments from P.Catinella and B.Edwards on RIE etching microelectronic devices.
One other question that I would like to direct to them or other interested parties would be the requirement for installing a "scrubber" on the exhaust fumes from the RIE. We are working up a estimate for the possible total cost of purchasing/operating/maintaining a plasma etcher in our laborato- ry. The requirement for a "scrubber" on the exhaust has been mentioned as a possibile environmental requirement. I have spoken to some manufactures and they don't use scubbers due to low flow rates in their machines and small quantities of gas involved.
Thanks again.
Richard Sartore US Army Research LAboratory AMSRL-EP-RA Fort Monmouth, NJ 07703 RSARTORE-at-MONMOUTH-ETDL1.ARMY.MIL
Dear Tamara Howard, I'm not sure if this is exactly what you are looking for but I have seen people in Jeremy Pickett-Heaps' lab at Melb. Uni. use teflon powder to coat glass slides before embedding between them. They don't actually grow the cells on the slides but just use them for thin layer embedding of algae in Spurr's. I'm not sure where teflon powder comes from - sorry. Can you grow the cells on plastic coverslips that you can section? I too have experienced having coverslips smashing instead of peeling nicely from the resin and actually found patience, practice and the gentle persuasion of a single-edged razor blade angled down between the glass and the resin to yield enough glass-free resin + cells to make it worth the trouble. David Orlovich.
Tamara, Try growing your cells on Permanox plastic dishes. The plastic is resistant to ethanol, acetone and resin and the polymerized resin is easily separated from the petri dish as long as the resin isn't too thick. I usually use a resin layer of about half a mm. If you need to use glass coverslips, you can try pushing the still-warm coverslip against a block of dry ice. The differential contraction rates will sometimes free the resin but it isn't 100% effective.
Rod Kuehn University of Minnesota
On Wed, 19 Jan 1994, Tamara Howard wrote:
} Does anyone have any experience with cell monolayers grown on glass coverslips? } I've seen a method reported where you coat the coverslip with carbon before } adding the cells; this is supposed to allow the resin to be stripped from the } glass for sectioning. Does it work? We thought the regular culture } substrates would allow release, but the glass just breaks when we try to "peel" } the resin away. Any suggestions would be VERY HELPFUL...I'm trying to section } glass, now. } Thanks! } Tamara Howard } Pitt Med/Pathology } Pittsburgh, Pa } tah-at-med.pitt.edu
} One other question that I would like to direct to them or other } interested parties would be the requirement for installing a } "scrubber" on the exhaust fumes from the RIE. } ...The requirement for a "scrubber" on the exhaust has been } mentioned as a possibile environmental requirement. I have spoken } to some manufactures and they don't use scubbers due to low flow } rates in their machines and small quantities of gas involved. } } Thanks again. } } Richard Sartore } US Army Research LAboratory } AMSRL-EP-RA } Fort Monmouth, NJ 07703 } RSARTORE-at-MONMOUTH-ETDL1.ARMY.MIL
The requirement for a scubber is more likely to be a matter of law and agency regs (military regs in your case) than real need, BUT it may also depend on what's coming off of the targets. The gas would be no trouble (so the manufacturers aren't going to worry about it), but if you're etching something like Gallium Arsenide semiconductors, there'll be reactive arsenic ions and the like coming out; that could be a problem. Scrubbing should be easily and cheaply done be bubbling the exhaust through distilled water (with maybe cotton batting in the outlet to make sure), which would be then disposed of by the usual regs for chemical waste. Phil Oshel po-at-parmly1.parmly.luc.edu
Re} TEM biol. cells on glass Tamara, I assume that you must embed the cells as a monolayer so that you can either orientate them or locate specific cells. If so, then you can take any of the blocks that you have already embedded, place them in liquid nitrogen and then warm them up. Repeated cycles of cooling and warming will crack the glass and will often cause it to shoot off the plastic. You must remove all resin from the top side of the block beforehand and be careful that the glass pieces do not get into your eyes. We use this method routinely to locate, and section, single cells that have been microinjected. The cells are grown on a locator slide and the pattern is transferred to the resin. You can help the glass removal by scoring the coverslip with a diamond before cooling and do not worry if the resin breaks. You will usually be able to find what you want amongst the pieces.
Cutting plastic coverslips is not easy, but a viable alternative to consider, if you only want orientation, is to grow the cells on the special filters produced by Costar and Falcon. These embed well and can be sectioned in resin. They are more difficult to cut cryosection from, but even this is possible.
If you only want a pellet of cells then it is better to grow them in plastic dishes. You can remove them, before fixation, by treating them with proteinase K, and after fixation by scraping with either a soft wooden or teflon scraper (not the normal cell scrapers that are available).
Good Luck
Paul Webster Yale School of Medicine (203) 785 5072.
Tamara Howard asks about stripping processed cells off glass coverslips, and suggests using carbon coated coverslips.
I have 3 suggestions:
1. I seem to recall seeing a paper/idea/suggestion once somewhere (???...) that rapid temperature changes at the glass resin interface will help break off the coverslip cleanly. Place a solid (pre-filled with resin and then polymerized) gelatin/Beem capsule over the cells on the coverslip and then polymerize them, attaching the cells to the capsule. Later, rapidly cool the coverslip, twisting the beem capsule. Play around, and let me know what works ! *** ONE ADVANTAGE: You could also first polymerize the cells between 2 coverslips (the one that it was grown on, and the other greased with vaseline so that it can easily be disloged after polym.), so that you could observe the cells clearly under phase contrast, mark the cells of interest, and finally place a solid resin capsule over the marked area, thus selecting the cells of interest.
2. Use propylene oxide (PO). This works like a dream when processing cells in plastic culture dishes: do all your processing for TEM right up to full dehydration with ethanol in the dishes. Monolayers process (fixation and dehydration) very quickly, and so it is a real easy technique to try. Decant the 100% ethanol from the dish, then quickly pour on pure PO, gently tilt the dish once or twice, and as the plastic of the dish starts to dissolve, the whole monolayer "peels off" and can be decanted into a suitable tube or small vial. Replace PO with the first change of resin, and carry on, embedding the mat of cells, perhaps using gentle centrifugation for the pure resin changes. Dave Sanan, now somewhere in California (give me a call, man!) first showed me this trick.
Perhaps the PO could even lift monolayers off the glass ? Would certainly work for plastic coverslips.
3. Hydrofluoric acid. Here's the real McCoy! Ref: J. Microsc. 104: 205- 207. Use hydrofluoric acid to etch away the coverslips, leaving the resin and cells behind. Be real careful as HFA is very corrosive. Work in a polyethylene or polytetrafluoroethylene dish in a fumehood.
Good luck
Ian
*********** ************ Dr Ian S Harper Int. Tel #: 27-21 938 0347 Experimental Biology Programme Int. Fax #: 27-21 938 0456 Medical Research Council Internet: iharper-at-eagle.mrc.ac.za PO Box 19070 Tygerberg, 7505 South Africa *****************************************
Thanks to B.Miner for forwarding the useful comments from P.Catinella and B.Edwards on RIE etching microelectronic devices.
One other question that I would like to direct to them or other interested parties would be the requirement for installing a "scrubber" on the exhaust fumes from the RIE. We are working up a estimate for the possible total cost of purchasing/operating/maintaining a plasma etcher in our laborato- ry. The requirement for a "scrubber" on the exhaust has been mentioned as a possibile environmental requirement. I have spoken to some manufactures and they don't use scubbers due to low flow rates in their machines and small quantities of gas involved.
Thanks again.
Richard Sartore US Army Research LAboratory AMSRL-EP-RA Fort Monmouth, NJ 07703 RSARTORE-at-MONMOUTH-ETDL1.ARMY.MIL
---------------------------------------------------------------------------- ---------------------------------------------------------------------------- --------------------------------------- Richard, If you are only dealing with a low volume of gas and you have a fume hood near by then you can do as we do exhaust fumes through there as most fume hoods, I would imagine, have a scrubber attached.
we only use plasma etching techniques for "deprossessing" integrated circuits for failure analysis so our gas flow is minimal.
For embedding monolayers we have been growing cells on a polymer film in place of glass cover slips. It is called Aclar and is available (in large quantities only) from ProPlastics, Linden NJ. It is optically very clear and separates easily from all embedding resins. It is apparently a Teflon- like substance. I was required to purchase more than a lifetime supply, so anyone who would like to try a sample should contact me. Not all cell lines will grow on the naked stuff. Some require a collagen coat and others never grow at all as they do on polystyrene dishes. or glass.
FROM THE PRESIDENT OF THE ROYAL MICROSCOPICAL SOCIETY
Something for Nothing! Bursaries Bursaries Bursaries ************************************************************************ Over many years it has never ceased to amaze me that RMS bursaries, to help eligible people attend meetings or courses, are not snapped up. It has even been known for them not to be fully taken up by the date of the sponsored event. Although admittedly the value of the bursary may not cover all of the expenses incurred in attending an event, they can be very useful primers in persuading other sponsors to make a contribution. Anyway, we announce here, bursaries for events in 1994.
RMS International Bursaries in Microscopy ************************************************************************ The RMS International Bursaries are intended to help young microscopists working outside Western Europe to attend RMS Courses or Conferences. The awards will be made to help with the registration and accommodation costs but it will not normally be possible to help with the cost of travel to the United Kingdom. The Society will offer up to six Bursaries annually and it is unlikely that any single award will exceed 250. There are no strict rules or definite age limits but it is likely that they will be made to assist young scientists who would otherwise be unable to attend the Course or Conference. The Bursaries are not limited to Fellows or Student Members of the RMS, but it is unlikely that an award will be made to an Applicant currently working in North America or Japan.
The application for an RMS bursary can be made at any time, but should be made as far in advance of the Course or Conference as possible. The application should include details of the Course or Conference to be attended, a copy of any abstract(s) to be submitted and also a copy of the Applicant's Curriculum Vitae and publication list (if appropriate). The application should be accompanied by a letter of support from the Applicant's Head of Department or from a Fellow of the Royal Microscopical Society.
It is expected that the Applicant will have made efforts to find funding from elsewhere and he/she will be expected to show that such an application has been made - even if it was not successful.
RMS Bursaries in Microscopy ************************************************************************ The RMS bursaries will only be available to Fellows or Student Members of the Society. Non-members of the RMS are not eligible. The awards will be made to help with the registration costs of a Course or Conference, but it will not normally be possible to help with the cost of travel or accommodation. There are no definite age limits, but it is likely that they will be made to assist young microscopists who would otherwise be unable to attend. It is unlikely that any single award will exceed 250 nor be made to an Applicant currently working in North America or Japan.
The application for an RMS bursary can be made at any time, but should be made as far in advance of the Course or Conference as possible as funding may be limited and to allow for fair refereeing of the applications.
It is expected that the Applicant will have made efforts to find funding from elsewhere and he/she will be expected to show that such an application has been made - even if it was not successful.
Applicants are advised that preferential consideration will be given to DipRMS and TechRMS candidates and those who have no financial support from their employer or other source. Please note that each application must be endorsed by the Applicant's Head of Department or employer.
The names of the successful Applicants will be published in the RMS Proceedings.
Application Forms for RMS Bursaries are available from: The Administrator, Royal Microscopical Society, 37/38 St Clements, Oxford OX4 1AJ, United Kingdom. Completed applications must be returned to the Administrator at the same address.
The Summer School in Light Microscopy for 1994 will be organised on a modular basis, as in 1993, to allow flexibility in matching the needs and interests of participants to the subject matter provided, and to make it possible to include minor or specialist topics for which the demand might not justify the provision of separate courses. These topics can be offered in conjunction with others and thereby share the availability of instructors and expensive up-to-date equipment. There will be evening lectures and discussions to obtain the best use of the time available, and the numbers of participants will be strictly limited.
The first three days (Sunday evening to Wednesday evening), the Principles of Light Microscopy module will consist of lectures, demonstrations and practical classes on the fundamental aspects of light microscopy and the various imaging and contrast modes which can be used in the observation and characterisation of biological specimens, polymers, ceramics, minerals and metals. This module will cover the basic concepts of all forms of microscopy, leading on to the functioning and limitations of the light microscope, and introducing techniques for enhancing contrast. Practical work will make use of a variety of types of microscope from the major manufacturers, and the module will provide a practical understanding of the phenomena which lie behind the principles and practice of light microscopy.
From Thursday morning, the course will be divided into three specialist modules from which participants may select. Analytical and Applied Microscopy is an extension of the Principles module, these two together covering approximately the same ground as our former one-week Principles course. It will build on the topics covered in the Principles module and will show how they may be applied in practice. This will assist the development of a systematic and analytical approach to microscopy. A workshop format will enable participants to examine and discuss the images obtained from samples which will be provided and their own specimens, using a wide variety of techniques. The emphasis will be on the correct adjustment of the instruments, the strengths and weaknesses of each technique and the interpretation of images. This module will be especially valuable for microscopists working in industrial laboratories.
The Polarised Light module will address itself to the interpretation of contrast, not only in ceramics and minerals, but also in biological materials in which components of the structure are birefringent. It will attempt to explain by the use of simple diagrams and demonstrations, and with the minimum use of mathematics, the colour changes which are observed, and how contrast may be enhanced by the use of accessories. This module is designed to be of use to biologists, materials scientists and geologists.
In the Image Recording module it will be assumed that participants have a thorough understanding of the techniques of imaging and contrast enhancement. The module will include the principles of photography and video imaging, the transfer of the image from the microscope to the camera, and the design and construction of image-recording equipment. Time will be available for exposing black-and-white and colour film, which will be processed and evaluated before the end of the module.
Participants may register for the whole week (for the Principles module and one of the three specialist modules), for the Principles module alone, or for one of the specialised modules alone. Because the specialised modules are designed to build upon the groundwork presented in the Principles module, we expect that participants in specialised modules will normally either attend the Principles module at the beginning of the week, or have attended a previous RMS Light Microscopy Summer School.
We anticipate the customary extremely generous provision of equipment and materials from the manufacturers, for all parts of the course.
Principles of Light Microscopy Sunday evening 17 July to Wednesday evening 20 July 1994 (a 3 day module)
This module will consist of lectures, demonstrations and practical work on:
The history of microscopy. Introduction to microscopy. Limitations of the eye. Resolution, Contrast, Magnification. Interactions between light and matter. Refraction and lenses, geometrical optics, conjugate planes. Aperture. Illumination of the specimen in transmitted and reflected light.
Lens aberrations and their correction; the choice of optical components. Diffraction and its consequences for the microscope image. Generation of contrast. Introduction to bright-field, dark ground, phase contrast, polarized light, differential interference contrast and fluorescence.
This module will adopt a workshop approach and consist of short informal lectures, demonstrations and practical sessions. It will cover the correct adjustment of the microscope, the strengths and weaknesses of each technique, and the interpretation of images, in both transmitted and reflected light. Participants will have the opportunity to become familiar with techniques of their own choice according to their interests, using where possible, their own specimens. Facilities for specimen preparation will, however, be limited. We expect equipment for the following techniques to be available:
This module is designed to introduce biologists, materials scientists and geologists to the usefulness of polarized light techniques, and will involve the minimum use of mathematics. Lectures, demonstrations and practical work will cover the following topics:
The nature of light and polarization. The interaction of birefringent materials with plane-polarized light. Stress optical effects, the use of accessories and compensating plates. Minerals, ceramics, biological materials and fibres. Reflected polarized light techniques. Introduction to conoscopic techniques.
This module will discuss the recording of images by drawing, by photography and by video techniques. Equipment and materials will be available to enable participants to record images in black and white and colour of their own specimens and to discuss difficulties and results. The following topics will be covered:
Drawing, photography and video methods; an overview. Transferring the microscope image to the film and video camera. Monochrome and colour film; basic principles, use and processing. Negative and reversal films. Printing. 'Instant' monochrome and colour films. Colour temperature, light sources, films and filters. Equipment for photomicrography; focusing and determining exposure. Photomacrography. Video cameras, monitors and printers.
TO OBTAIN FURTHER DETAILS OF THIS COURSE SEND AN EMAIL MESSAGE WITH YOUR NAME AND ADDRESS TO RMS-at-UK.AC.OX.VAX, OR CONTACT THE RMS ON TELEPHONE +44 (0)865 2488768, FAX +44 (0)865 791237.
A range of techniques have been devised for localising antigens at the electron microscope level. One of the difficulties commonly faced is the choice of which method to use with any particular antigen-antibody combination. The aim of the course is to provide a theoretical and practical introduction to the various methods available and covers antigen location, using both transmission and scanning electron microscopy.
The advantages and disadvantages of the various methods available will be discussed, but the emphasis will be on the practical aspects of the techniques and will provide experience of the following methods: pre-embedding labelling for scanning electron microscopy; low temperature embedding in Lowicryl resins; preparation and labelling of both resin sections and thawed cyrosections; rapid freezing by impact and high pressure followed by freeze substitution and low temperature embedding; silver enhancement of colloidal gold for light and electron microscopy; immunolabelling of 1ęm resin sections; and epi- polarised light microscopy. Registrants are encouraged to discuss specific problems with the course organisers and if possible, to bring samples with them for processing and labelling during the practical sessions, which will be supported by Leica (UK) Ltd.
The course is primarily aimed at electron microscopists with experience of routine processing methodology who wish to become familiar with ultrastructural immunocytochemical labelling techniques.
The practical nature of the course, means that numbers will be restricted to a maximum of 10 registrants.
FOR FURTHER DETAILS CONTACT THE RMS - TELEPHONE +44 (0)865 248768, FAX +44 (0)865 791237, EMAIL RMS-at-UK.AC.OX.VAX.
The unprecedented upsurge in the application of immunocytochemistry in the life sciences during the past decade continues with increasing vigour. Both light and electron microscopy are important techniques in routine diagnosis and research in medicine and biology. This technology has contributed so much towards our current understanding of the cell, that the modern microscopist must be part immunologist as well as being skilled in microscopy.
The underlying principles of immunocytochemistry apply equally to light and electron microscopy. This five-day course has been specially designed to utilise this overlap and therefore, will be of value to both light and electron microscopists. The course is structured towards a technical appreciation of immunocytochemical techniques, since once they are mastered, they can be applied to any system. Each year this popular course is updated in the light of new developments in immunocytochemistry and is also suitable for participants interested in the plant sciences, as we teach some of the specialist techniques required for the handling of plant cells. The course will be of immense value to any life science microscopist/cell biologist of any background, who intends to use or is starting to use immunocytochemistry for routine purposes or research.
The course counts towards qualification for the Diploma of the Royal Microscopical Society.
The main emphasis of the week will be to give participants sufficient practical experience and knowledge of immunocytochemistry to carry out immunolabelling in their own laboratories. At the same time, a series of lectures will be given by more specialist exponents in various areas of immunocytochemistry.
The three practical days will be led by experts in their particular field: Tony Leathem and Susan Brookes (LM immunocytochemistry), Paul Monaghan (immunogold labelling for EM) and David Hughes (silver enhancement). After an introduction on the biology and production of antibodies, the practicals will be backed up with lectures on the applied aspects of the respective techniques. Of particular interest is the series of specialist lectures which will include, botanical immunocytochemistry (Chris Hawes), in situ hybridisation (John Davies), confocal microscopy (Mark Fricker), immuno-scanning EM and special applications of colloidal gold. An evening workshop will be held to introduce cryo-techniques in immunocytochemistry and to demonstrate some of the latest equipment used in low temperature tissue preparation.
FOR FURTHER DETAILS CONTACT THE RMS - TELEPHONE +44 (0)865 248768, FAX +44 (0)865 791237, EMAIL RMS-at-UK.AC.OX.VAX.
Last year we experimented with a new format for our Flow Cytometry course. Following its success both in attracting students and instructing and informing them, we will continue with the new format. There will be two courses - basic and advanced - run sequentially to give the potential student the choice of attending either or both of the courses. We anticipate that there will be at least three bench top cytometers for use by the students and one machine equipped with two lasers for the advanced course. As usual, we will rely on the generous co- operation of the manufacturers, Becton-Dickinson, Coulter and Ortho, who lend us both the machines and experienced operators to run them.
The basic course will assume little prior experience and take the student through the most important applications. Although it will fill the needs of someone working in a research environment, it will have a slight clinical bias. The majority of bench top flow cytometers are now to be found in clinical laboratories.
The first day will consist of a series of talks describing the basics of flow cytometry. A simple practical (measurement of a DNA histogram from cultured cells) will serve as an introduction to using the bench top flow cytometers.
There will be two practicals on the second day. In the first, students will use antibodies labelled with three different fluorochromes to identify lymphocytes subsets in human peripheral blood. This practical will also demonstrate the importance of using light scatter to separate lymphocytes, monocytes and granulocytes in samples of blood. In the second practical (lead by Richard Camplejohn from St. Thomas's Hospital), nuclei will be extracted from a formalin-fixed, paraffin embedded tumour and the DNA histogram recorded. The rest of the programme will include lectures on the analysis of DNA from clinical samples and on immunophenotyping in a hospital laboratory.
The third and final day will also be the first day of the advanced course. In the practical demonstration, lead by George Wilson (GRC Gray Laboratories), students will investigate the cell cycle kinetics of a mouse tumour which will have been labelled in vivo with a thymidine analogue (bromodeoxyuridine, BrdU). There will be a lecture on applying this technique and its clinical application and on DNA measurement and cell cycle analysis, the principles of cell sorting and the measurement of antigens associated with cell proliferation.
The advanced course will join us for the last day of the basic course (see above). On their second day, they will prepare and analyse chromosomes, using both bivariate and univariate analysis. We will set up a flow cytometer in the lecture theatre and project the computer screen so that it can be seen by the whole class. Jim Watson (MRC, Addenbrookes, Cambridge) will lecture on time as a parameter and then run experiments in the lecture theatre on intracellular enzyme kinetics looking at esterases and glutathione-S-transferase. A lecture on the measurement of intracellular pH and calcium ions will also be followed by a live demonstration. The other lectures will be on chromosome analysis and sorting and on further applications in cell and molecular biology.
The third and final day will consist of lectures on studying apoptosis, measuring multi-drug resistance in tumours and lectures and practical demonstrations on the measurement of cell cycle kinetics using the BrdU-Hoechst/PI method and on the interaction of fluorochromes and DNA.
We expect that anyone attending our basic course will come away with a sound grasp of the principles of flow cytometry, an understanding of the concepts behind data analysis, a feel for some of the problems and the confidence to run the commoner applications. Everything in the course will be relevant to workers in a clinical environment.
The advanced course will be at the forefront of the technology. It should give students a broad understanding of the wide range of applications of flow cytometry in a modern research laboratory. It will help them to establish new techniques and ought to give them ideas for new experiments in their own laboratories .
We hope that many students will take the opportunity to stay the whole week and to benefit from a broad look at flow cytometry - from basics to the most advanced applications. The RMS course is the only comprehensive course on flow cytometry to be run in the UK
FOR FURTHER DETAILS CONTACT THE RMS - TELEPHONE +44 (0)865 248768, FAX +44 (0)865 791237, EMAIL RMS-at-UK.AC.OX.VAX
We have versions of Diffract, DiffractII, and Desktop Microscopist. The best one is the Desktop Microscopist by far. It works much better, doesn't have as many faults as diffract and is more user friendly. It doesn't have that annoying propensity to crash all the time as diffract.
I would like to use freeze substitution for a low magnification TEM study of cell-extracellular matrix relationships in amphibian embryos. Can anyone suggest an appropriate fixative and concentration for adding to the sustitution medium (MeOH) prior to embedding in EPON?
Dave Parichy Section of Evolution and Ecology Univ. of California, Davis 916 752 3634
Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii" To: "Rick A. Harris" {szrick-at-bullwinkle.ucdavis.edu}
} I would like to use freeze substitution for a low magnification TEM study } of cell-extracellular matrix relationships in amphibian embryos. Can } anyone suggest an appropriate fixative and concentration for adding to } the sustitution medium (MeOH) prior to embedding in EPON? } } Dave Parichy } Section of Evolution and Ecology } Univ. of California, Davis } 916 752 3634
You might find useful information in:
Allanspach, A. 1993. Ultrastructure of early chick embryos after high pressure freezing and freeze substitution. Micoscr. Res. Techn. 24:369-384.
Hippe-Sanwald, S. 1993. Impact of freeze substitution on bioolgical electron microscopy. Microsc. Res. Techn. 24:400-422.
Hunziker, E.B. 1993. Application of cryotechniques in cartilage tissue preservation and immunoelectron microscopy: potentials and problems. Microsc. Res. Techn. 24:457-464.
McDonald, K. and M. Morphew. 1993. Improved preservation of ultrastructure in difficult-to-fix organisms by high pressure freezing and freeze substitution: I. Drosophila melonagaster and Strongylocentrotus purpuratus embryos. Microsc. Res. Techn. 24:465-473.
I have just had a look in my favourite freeze-substitution book (Cryotechniques in Biological Electron Microscopy, eds: R A Steinbrecht and K Zierold) and they suggest the following freeze-substitution medium for methanol substitution:
Methanol containing 0.5% uranyl acetate, 1% OsO4, 3% glutaraldehyde and 3% water (the water comes from the 50% glutaraldehyde they used).
It actually looks a bit complex to make the solution up - they suggest:
add 9 mL of 50% aq. glutaraldehyde (in a liquid nitrogen precooled flask) to 60 mL methanol, then 3 mL of a 20% (w/v) soln of uranyl acetate in methanol are added; in a second precooled flask 1.5 g osmium tetroxide are dissolved in 75 mL methanol; both flasks are cooled to about 220 K, their contents poured together and vigorously shaken.
They say that the mixture is highly reactive even at 240 K and so should be used in a few hours.
Substitution time was 8 hours each at -95 deg C, -60 deg C and -30 deg C. Then 30 minutes at 0 deg C. Replace the substitution mix with pure acetone (still at 0 deg C), warm to 7 deg C and infiltrate with araldite/Epon.
The original reference to this protocol is: Muller, M., Marti, T., and Kriz, S. (1980). Improved structural preservation by freeze-substitution. In: Brederoo, P., and de Priester, W. (eds). Electron Microscopy 1980, vol.II. Proc. 7th Eur. Congr. Electron Microsc., Leiden, pp. 720-721.
I've always used 2% OsO4 in acetone at -70 deg C for up to 7 days and then embedded in Spurr's resin. The best reference I have for that is:
Howard RJ and O'Donnell KL (1987). Freeze-substitution of fungi for cytological analysis. Exp. Mycol. 11:250-269.
I hope this is of some use.
David Orlovich School of Biological Science University of NSW PO Box 1 Kensington NSW 2033 Australia
Subject: Time:2:18 PM OFFICE MEMO RE} SEM RES STD Date:1/24/94 The method I have used with good success was published on p. 68 of the May 1987 issue of the EMSA Bulletin. If you don't have access to that issue of the Bulletin, send me your FAX address and I'll send a copy to you. Bigelow-at-umich.edu.
I am looking for software (vendor / public domain) that can draw crystal structures and print these diagrams on a laser printer. I would like to be able to enter the crystal system, cell edges, etc.. and have the diagram show the coordination of differing cation sites. Any suggestions?
Message-Id: {MAILQUEUE-101.940125090324.512-at-FS-IAM-1.JRC.NL} To: Microscopy-at-anlemc.msd.anl.gov
I intend to do some internal strain measurements using HOLTZ lines, so could anyone suggest any software which allows the simulation of HOLTZ lines, and where I could find it?
Kodak has discontinued direct duplicating film 2468 in 100 foot rolls. We have used this film successfully for years to make presentation slides from halftone prints. What should we use now as a replacement?
Received: from MAILQUEUE by MICROBIO1 (Mercury 1.11); Tue, 25 Jan 94 11:24:30 EST Return-path: {EMLAB-at-opus.mco.edu} Received: from emoryu1.cc.emory.edu by transporter.microbio.emory.edu (Mercury 1.11); Tue, 25 Jan 94 11:24:27 EST Received: from anlemc.msd.anl.gov by emoryu1.cc.emory.edu (5.65/Emory_cc.3.4.12) via SMTP id AA15050 ; Tue, 25 Jan 94 10:51:32 -0500 Return-Path: EMLAB-at-opus.mco.edu
Kodak has discontinued direct duplicating film 2468 in 100 foot rolls. We have used this film successfully for years to make presentation slides from halftone prints. What should we use now as a replacement?
KODAK OFFERS A FILM CALLED " RAPID PROCESS COPY " (CAT. NO 174 6031 FOR THE 150 ft ROLL) WHICH IS A DIRECT REVERSAL FILM DESIGNED FOR THE PURPOSE YOU DISCRIBE. THIS FILM IS QUITE SLOW : EXPOSURES IN THE 30 TO 45 SECOND RANGE AT f 4.0 ON MY SETUP. ONE NEEDS TO CALIBRATE THEIR COPY STAND SETUP TO THE FILM AND PROCESSING.
THE PROCESSING IS VERY SIMPLE: DK 50 DEVLOPER FOR 10 MINUTES, FOLLOWED BY STOP AND FIXER.
I HAVE BEEN USING THIS FILM SINCE THE MID- EIGHTIES WITH EXTREMELY GOOD RESULTS. ANY QUESTIONS? CALL ME. lmelsen-at-unix.cc.emory.edu
Via: uk.ac.birmingham.computer-centre.ibm3090; Tue, 25 Jan 1994 16:27:15 +0000
On Tue, 25 Jan 1994 MACLAREI-at-ibm3090.computer-centre.birmingham.ac.uk wrote:
} I wonder if anyone has any good ideas on how to electropolish commercial } purity Titanium without getting precipitation of hydride needles. } } I have tried using a variety of acid based solutions, most of which } contained perchloric acid in varying concentration. All of these, however, } result in hydride precipitation. } } The net result of all of this is that I find it very difficult to prepare } decent specimens of Titatium by electropolishing. If anyone has any good } ideas as to how these problems can be overcome I would be very glad to hear } from you. } } Ian MacLaren
Ian,
You may want to try the following, (although I have NOT tried this particular recipe):
Ethanol (96%)................90 ml n-Butyl alcohol..............10 ml Aluminum Chloride.............6 g Zinc Chloride................28 g for 1-6 minutes; 20 -25 V dc; stainless steel cathode; room temp.; need to keep agitated (the solution, NOT the user {grin!} ) to prevent a passivating layer from forming: can use a stirrer, or oscillate the anode quite rapidly at a fixed distance from the cathode (approx. 1 to 2 cm). * remember: nicely TOXIC!!! {ref.: The Electrolytic and Chemical Polishing of Metals; Tegart; 1959; Pergamon Press}
Hope this is helpful, Rob
X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X X Rob Tayloe X MSM Spelunkers Club X X Metallographic Lab. X Missouri Speleological Survey /-v-\ \-v-/ X X Rolla Research Center X Bat Conservation International X X U.S. Bureau of Mines X Missouri Cave & Karst Conservancy \-v-/ X X tayloe-at-rorc.usbm.gov X National Speleological Society #32993 /-v-\ X X (314) 364-3169 x247 X American Cave Conservation Association X X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X\X
I think there is new chemistry for using T MAX films for direct positives. Someone from Kodak should pick up on this message and let the rest of us know about it. ***************************** * Greg Erdos * * Director, ICBR EMCL * * University of Florida * * Gainesville, FL 32611 * * gwerdos-at-gnv.ifas.ufl.edu * * 904-392-1295 * *****************************
I called my supplier of 2468 today. They say that the film has NOT been discontinued. (And this company claims they are the only E. Coast supplier!) What's the truth???
If your prints come from large-format negatives, you can photograph the negatives on a light box with technical pan or t-max. If you need a direct positive film, you can use Ektachrome color slides or use t-max with a direct-positive developing kit.
Rod Kuehn University of Minnestota
On Tue, 25 Jan 1994 EMLAB-at-opus.mco.edu wrote:
} Kodak has discontinued direct duplicating film 2468 in 100 foot rolls. } We have used this film successfully for years to make presentation } slides from halftone prints. What should we use now as a replacement? }
Return-path: {BERGRH-at-MSUVX1.MEMST.EDU} Received: from memstvx1.memst.edu by gnv.ifas.ufl.edu (PMDF V4.2-11 #3240) id {01H848IIDF408WW4DH-at-gnv.ifas.ufl.edu} ; Tue, 25 Jan 1994 23:26:34 EST Received: from MSUVX1.MEMST.EDU by MSUVX1.MEMST.EDU (PMDF V4.2-14 #5958) id {01H845ET8EPC9BWPOF-at-MSUVX1.MEMST.EDU} ; Tue, 25 Jan 1994 22:28:15 CST
Greg-- I am reading your Microscopy post and, not adept at replying to these public forums, am writing you direct. I used the TMax direct reversal kit for my talk at this summer's MSA meeting and was very pleased with the quality of the slides it produced. I made "superslides" by using (hard to find) 127 film mounts and TMax 100 film size 120. The kit specifies rating the film at ASA 50, half its normal speed. The kit instructions are well written and I got good results from the first roll on. Instructions for extending development with subsequent rolls are clearly written--as I recall the kit will process about 8 or 10 rolls and cost me $35 (?). Significantly more rolls of 35mm would be possible
Direct Pos. Film users: Kodak makes a TMax direct postive film developing kit for making black and white slides. Catalog number 812 1188. Cost: approx. $30. When doing line copy I have also used LPD4 film for continuos tone copy and have had very good results. There's a little trickery involved but it allows you to do line and continuous copy on the same roll of film. If anyone is interested, I'll tell how it is done. Phil Rutledge prutle1-at-gl.umbc.edu
Message-Id: {MAILQUEUE-101.940126090559.480-at-parmly1.parmly.luc.edu} To: microscopy-at-anlemc.msd.anl.gov
} Date: Wed, 26 Jan 1994 09:26:33 -0500 (EST) } From: rutledge phil {prutle1-at-gl.umbc.edu} } Subject: Tmax } To: microscopy-at-anlemc.msd.anl.gov
} Direct Pos. Film users: } Kodak makes a TMax direct postive film developing kit for making black } and white slides. Catalog number 812 1188. Cost: approx. $30. When doing } line copy I have also used LPD4 film for continuos tone copy and have had } very good results. There's a little trickery involved but it allows you } to do line and continuous copy on the same roll of film. If anyone is } interested, I'll tell how it is done. } Phil Rutledge } prutle1-at-gl.umbc.edu
I'll second this--I've used LPD-4 for making direct-positives of photographs for slides when I only want to mess with developing one type of film. 8 secs. at 1/2-stop intervals from f111/2 to f4 or 41/2 will usually get you a usable frame. This range of f-stops is conservative; actually shouldn't need more than a couple of brackets once you figure out your local conditions. The Direct Positive sold by Ted Pella does do a better job, but.... Phil Oshel
Kodak make a T-Max Reversal Kit for direct positives. The catalogue number is K-8121188. It comes as a 1 quart unit. In Canada it sells for about $35.00.
Kodak make a T-Max Reversal Kit for direct positives. The catalogue number is K-8121188. It comes as a 1 quart unit. In Canada it sells for about $35.00.
} } I have found something that I think works better, but takes a day to } develop - regular color Ectachrome slide film (Tungsten). The results } are very good and having somebody else fool with the wet chemistry is } much better. I think that the development process is C-47 which is the } same for color negatives which means that you can have it done at 1 hour } photo shops. As a reult, I only use the MP 5360 when I have an } emergency. } } } Scott Walck } walcksd-at-ml.wpafb.af.mil } Materials Directorate } Wright Patterson AFB, OH
I have had excellent results with Ektachrome, but find it takes 25-30 minutes to development: process E-6 with the Kodak hobbyist pack, 10-30 minutes to dry, depending on if you have a film drier. Most 1-hr photo-shops that I know of won't do E-6 films. Phil Oshel po-at-parmly1.parmly.luc.edu Parmly Hearing Inst. see c. v. on MSA Bulletin Board. Please!
Slides I can't believe that anyone is satisfied with the 35mm format for projection slides. Besides, copying prints with a camera always results in a loss of quality. The best way is to take the information straight from the negative, although this does not let you label the image with letraset. I print directly onto film, using an enlarger and mount the image in the super-slide frames (40x40mm) loved by electron microscopists and hated by projectionists. The image can be carefully framed and the contrast can be easily manipulated. The film to use is Agfa sheet film, so you will have to find your own supplier, and can be purchased in boxes of either 10x8 or 10x12. Use it as you would paper under an enlarger. For a soft image use "Litex premium camera film 0910P". For a harder finish use "Litex camera halftone film 0811P". These films are normally developed in Gevaline G7C developer, also from Agfa, which will produce high contrast but they can also be developed in D-19 for a softer contrast. I am sure that acutance developers will also work well. The film can only be handled in red light and has to be dish developed but the results are worth it. Try it and compare with camera copies. There is nothing like a big image to make your point. If you are interested and require more details then feel free to contact me. Paul Webster Yale School of Medicine paul_webster-at-quickmail.yale.edu (203) 785 5072.
} Slides } I can't believe that anyone is satisfied with the 35mm format for projection } slides. Price! Money! Grant funds! 35mm is cheap! And everyone has a slide projector that will take 35mm. Phil Oshel
I would appreciate any feedback from users of Digital confocal software packages like MICRTOME from VayTek or comparable systems.
1) Does cost justify capabilities.
2) How it compares to laser systems?
3) Are manus userfriendly?
4) If you used package already would you buy an upgrade or move on to a different system?
Thanks.
Cesar D. Fermin, Ph.D. Tulane University Medical School Department of Pathology 1430 Tulane Ave/SL79 New Orleans, La 70112-2699 (504) 584-2521 Fax 587-7389
"Price! Money! Grant funds! 35mm is cheap! And everyone has a slide projector that will take 35mm. Phil Oshel"
What a rude reply Phil! I don't deserve that.
Most EM labs have a darkroom with an enlarger. A box of 100 sheets of film is cheap and will last for years (I bought my boxes four years ago and still have enough to send to you, Phil Oshel, if you want to try out the system. The 40x40mm slide holders (from GePe) fit into a 35 mm slide projector -they just use up more of the available space. If anything it is all cheaper than buying a copystand and 35 mm camera. Why do I bother?
During my short stay in Sendai, Japan, I learned a good way to make overheads from electron microscope negatives. The material is FUJIGRAPH PROJECTION FILM PT-100, which comes at least in size 21x29,7cm (DIN A4) in 100 sheets packages. This film can be used just like the enlarging paper in the darkroom. The results are far better than with any oldfashioned way. These overheads are good for teaching purposes because you can easily make markings on them.
Regards, J M
- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - Jouko K. Maki Navigare necesse est... Laboratory Manager, Ph.D. Laboratory of Electron Microscopy Kiinamyllynkatu 10 FIN-20520 TURKU FINLAND University of Turku Tel.: + 358 21 633 7318 INTERNET: jouko.maki-at-utu.fi FAX.: + 358 21 633 7380
} Re} Slides } } "Price! Money! Grant funds! 35mm is cheap! } And everyone has a slide projector that will take 35mm. } Phil Oshel" } } What a rude reply Phil! I don't deserve that.
My apologies...but I repeatedly run into people, usually from medical schools, who don't understand why others don't use this or that, usually expensive, technology, instead of all the old stuff. The litany can get very frustrating. The answer is usually money. And often administrators. Labs and smally schools have photographic equipment & 35mm cameras, 35mm slide mounts, etc.. I have worked in EM labs for 13+ years, and have spent many hours all of the equipment you mention below. I have had much trouble with 40X40 slide holders jamming in 35mm projectors, but haven't tried the brand you recommend. Plus, multiple-image plates & 4"X5" Polaroid images still need to be photographed on the copy stand. And a good 35mm camera is still the cheapest, easiest way to produce the slides. Pros don't use them out of tradition. Personally, I'd prefer to use 120 film, or even 4X5, but....And most light microscopes come with 35mm camera backs. Meaning you need all that stuff anyway if you're a service lab, or anyway, not a dedicated lab where only one photpgraphic system is used (which most are). But what type of film comes in 100 sheet boxes & lasts for years? The film I've seen & used in TEMs & any other microscope--sheet, 35mm, or 120 roll--goes quickly. Printing paper's worse. Especially with students and EM courses! Thank you for the offer to send some materials, but it'd be a loss now--our grant & my job got cut, so it wouldn't be used. Phil Oshel } Most EM labs have a darkroom with an enlarger. A box of 100 sheets of film is } cheap and will last for years (I bought my boxes four years ago and still have } enough to send to you, Phil Oshel, if you want to try out the system. The } 40x40mm slide holders (from GePe) fit into a 35 mm slide projector -they just } use up more of the available space. If anything it is all cheaper than buying } a copystand and 35 mm camera. Why do I bother? } } }
To E'mers wanting EM slides: The easiest way to make EM slides is to put your neg in an enlarger and shoot onto another piece of EM film (type 4489). Develope it as you would normally process em film. This allows you to develope in a tray under a red safelight and allows control of the density of the slide. It gives great contrast in the slide or you can control the contrast by the way you develope the film. I started doing it this way for em slides only over 25 years ago and have always gotten good contrast whether the original negative was on the flat or contrasty side. Phil Rutledge prutle1-at-gl.umbc.edu
As I said in a previous message, I use LPD-4 for continuous tone slides on the same roll of film when I am doing line copy. The way I do this is as follows: I use a Polaroid MP-4 copy stand equipped with a Nikon F3 and a 90mm macro lens.
Line Copy __________
Exposure: 5 seconds/f:6 Develope: D-19 full strength- 2 minutes Fix: 2 minutes in rapid fix with Orbit Bath added. Orbit Bath allows a 2 minute fixing time and a 5 minute wash. It's better than Kodaks Hypo Clearing Agent. Wash: 5 minutes, dry, mount
Continuous Tone ________________
This requires a little playing with to determine your exposure. I use same exposure for the line copy but I add 3-5 seconds more. I pre-fog the film by exposing the copy for 8-10 seconds. At the same time I continuously move a grey scale card under the lens until I get to the 5 second mark for the line copy exposure. Process as above. I have always had good results with this method. You just need a little coordination exposing this way. Phil Rutledge prutle1-at-gl.umbc.edu
This is the text of an advertisement that will appear in the british press next week for a postdoctoral position which will become vacant on the 1st of June this year. We would like to identify an appropriate candidate as soon as possible - hence the short deadline. Applications will be accepted by FAX but NOT BY ELECTRONIC MAIL. Please note that, due to circumstances beyond our control, we will have to give preference to candidates who are citizens of nations in the European Economic Community.
The University of Birmingham School of Metallurgy and Materials
POSTDOCTORAL RESEARCH FELLOWSHIP
Applications are invited for the above post to work on a 2.5 -year SERC-funded project entitled "Mechanical Behaviour of Nb3Al Alloys". The successful applicant will investigate the basic defect structure and deformation mechanisms in these alloys and to explore strategies for enhancing their ductility.
A Ph.D. degree, extensive experience of electron microscopy techniques and familiarity with the physical metallurgy of intermetallic compounds are essential.
Preliminary enquiries should be directed to Dr. M. Aindow - Tel: (44) 21 414 5188, FAX: (44) 21 414 5232, Email M.AINDOW-at-BHAM.AC.UK, or Dr. I.P. Jones - Tel: (44) 21 414 5184.
Application forms and further particulars may be obtained from The Director, Staffing Services, The University of Birmingham, Edgbaston, Birmingham, B15 2TT, United Kingdom or telephone (44) 21 414 6483 (24 hours) and quote reference G10613/94. The closing date for receipt of applications is 18/2/94.
The University of Birmingham is an equal opportunity employer.
Mark Aindow, School of Metallurgy and Materials, Telephone; (021) 414 5188 The University of Birmingham, FAX; (021) 414 5232 Elms Road, Edgbaston, Birmingham, Email; M.AINDOW-at-BHAM.AC.UK GB B15 2TT, United Kingdom.
I have had quite a few requests asking how to access the new sci.techniques.microscopy newsgroup. So, I thought I would post a summary of the whats, wheres and hows of Usenet News, as I understand them, to the NIH-Image and Microscopy mailing list.
What is Usenet News? Usenet News, unlike email, is not distributed to an individuals account, but rather to a server at a particular institution (be it a government lab, university or private company) that can then be accessed by individuals at that institution. Usenet News is a system where messages are posted to newsgroups that are designed to cover a particular topic. Examples of topics are rec.windsurfing (recreational: windsurfing), comp.sys.mac.digest (Computer: Macintosh question and answer digest), sci.materials (Science: Materials Science) and, of course, sci.techniques.microscopy (Science: Microscopy Techniques of all kinds). There are many hundreds of worldwide newsgroups, i.e. groups that are distributed around the world, and then there also are groups that are only distributed locally. Examples of world-wide groups are those I have mentioned above and here at the University of Michigan we have local groups for class disscussion. As an example, for our electron microscopy course (MSE 562) we could have a group umich.eng.mse562 and it would only be available to readers in the umich.edu domain.
Postings to each newsgroup are stored on the institution's local NNTP (sorry guys I dont what this is an accronym for unless it is Network News Transport Protocol!) server and distributed to other sites. If the same kind of system is used now as was used when Usenet was first started, again I am no expert here - just a user, then each news server will exchange messages with a number of other servers geogrpahically close to it and the postings will sort of hop from one system to another and propgate around the world. When Usenet was first started the transfer was by dial-up modem lines, now the how system uses the Internet. If there is someone out there who can describe this process more exactly, please let me know or post a summary.
How do I read Usenet News and post to Newsgroups? If you want to use Usenet News you need access to an NNTP server and news reading/posting software to run on your local computer, workstation or terminal. You should contact your local network adminsitrator and ask them if you have access to a Usenet feed and which net-news software they recommend for the machine you use. If they do not have access, ask them why not! The functionality of each news program is best expalined by a local expert, I cannot possibly go into all of the details of even one news program here. Suffice to say however, if you have access to a news posting and reading program and access to the accompnaying NNTP server, you can post articles, questions and information to be read by people around the world.
Which software should I use? As I say it depends on you machine and what is available to you locally. I use a Mac and my favorite software is NewsWatcher (which is free, a definite bonus). The lastest verion of this software is available by anonymous ftp from ftp.acns.nwu.edu (the same place as Disinfectant, the antiviral utility for the Mac) in the directory /pub/newswatcher. The current version is 2.0d17. Ask around locally to see what you have available.
Why News and not just Email? The advantage of News over email mailing lists is the messages reside on a remote machine and do not clog up your mailbox. Some people have severely restricted mailbox size allocations and the output from a mailing list can swamp them and prevent them from receiving other mail. With Usenet you only download the News you want to read. You can subscribe to a small subset of the total newsgroups and selectively scan through those. You may view all of the subjects of the articles in a group before you read any of them. It is quite a flexible system.
I cant get to the Internet, what do I do? The one major complaint about Net News is that it is not accessible unless you have a connection to the Internet. The mailing lists can be accessed from Compuserve, America On-Line, GEnie, etc. An connection used to be an expensive proposition, however, these days it can be as reasonably priced as Compuserve or the other dial-up network services. For example "The World" (1-617-739-0202) charges $20 per month for 20 hours of connect time to Internet services. Another provider "The WELL" (Whole Earth 'Lectronic Link - 1-415-332-4335) charges $15 per month and $2 per hour for use. For a local provider please call the InterNIC Information Services at 1-800-444-4345.
This is just a brief outline that I have put together on the spur of the moment (as someone recently put it "its a stream of consciousness"!) Hope it helps. Drop me a line if I have made any major mistakes or left anything vital out of this message. Cheers, John Mansfield.
} 2) How it compares to laser systems? One potential problem is that to collect images using a cooled CCD camera the exposure times must be much longer than with a laser scanning confocal microscope. For instance, we did a comparison of a double labeled cultured cells scanned with a cooled CCD camera and with the BioRad MRC 600. With the CCD camera the Cy3 stain looked great (deconvolved with the quick mode of the Vaytek system). However, the FITC bleached before we could collect a few sections. On the BioRad, we were able to collect both signals simultaneously without significant bleaching of the FITC. Also, we looked at the BDS deconvolution system. We were very impressed with the deconvolved images, but here is a major difference between confocal and deconvolution: Using a confocal, investigators from all over the university can come in, scan their samples, immediately see results, get hard copy or send images to other computers instantly, and leave. If they were using deconvolution they would have to post-process every optical section before getting results. Personally, I would like to see how the BioRad confocal images would look processed with one of the deconvolution packages. -Michael Cammer
The problem "jjerome-at-isnet.is.wfu.edu (Jay Jerome)" mentioned as spectral effects when he mounted slides in glass mounts may be Newton rings. This problem may be eliminated by using anti-Newton glass in the mounts. Companies such as Wess or Gepe market this kind of glass for slide mounts. Contact your local photo shop or look in one of the photography magazines for advertisements.
} With regard to suggestion that our problem of spectral effects, we do use } Gepe anti-newton glass slides. However, the effect is reminiscent of the } effects one gets if you do not use anti-newton glass. It does not occur } however when we use thinner based film for making our slides. Only when we } use EM negative film such as 4489. Any other suggestions?
If you're not doing so already, you might try using a mask between the backing side of the film and the glass. This will eliminate the contact between the two flat surfaces that can produce Newton rings.
Way back when, we used to use commercially available masks cut from black paper. I've also used aluminum ones from EMDE Products, Inc. I don't even know whether they are still in business. The address in the literature I have from them doesn't even have a zip code. The address given is 2040 Stoner Ave., Los Angeles, CA.
John chandler-at-lamar.ColoState.EDU Fort Collins, CO
} Return-Path: {Vitor-at-dsif.fee.unicamp.br} } From: Vitor-at-dsif.fee.unicamp.br } Date: Wed, 26 Jan 94 09:26:00 EDT } To: venema-at-dutentb.et.tudelft.nl, Snitka-at-dsif.fee.unicamp.br, } tprohas-at-email.tuwien.ac.at } Subject: CALL FOR PAPERS } } } } } ----- Begin Included Message ----- } } } From PHOTOEXE%ILCTEHOL-at-VMS.HUJI.AC.IL Sun Jan 23 10:06:12 1994 } Date: Sun, 23 Jan 94 09:59 GMT } From: INTL BULLETIN ON PROCESSES AND APPLICATIONS } {PHOTOEXE%ILCTEHOL-at-VMS.HUJI.AC.IL} } Subject: CALL FOR PAPERS } To: VITOR-at-dsif.fee.unicamp.br } Content-Length: 1791 } } FRONTIERS IN SCIENCE AND TECHNOLOGY } AT NANO-MICRO SCALE } } July, 28-29 - 1994 } Guaruja - Sao Paulo State - Brazil } } Topics: } (not limited to) } } Scaling Laws - Theory } Scanning Probe Microscopies (STM, AFM,....) } Sensors } Piezoelectric Drivers } Machining (Beam, Plasma, Molecular, ...) } New Materials (Diamond-like, Porous Silicon....) } Synthesis and Structures (Biology, Microelectronics....) } Computation } ..... } } Abstract Deadline March, 20, 1994 (two printed A-4 pages) } Manuscripts at the Conference Site } } Organizing Comitttee } ==================== } } } Prof. Vitor Baranauskas - State University of Campinas - Brazil (chairperson) } Prof. Valentinas Snitka - Vibrotecnika - Lithuania } Prof. Ioshiaki Doi - State University of Campinas - Brazil } Prof. Aaron Peled - CTEH - Israel } Dr. Vladimir J. Trava-Airoldi - Instituto Nacional de Pesquisas Espaciais -Brazil } Prof. Gilberto Mattos Gualberto - State University of Campinas - Brazil } } } Organized by: } ============ } Sociedade Brasileira de Vacuo - Brazilian Vacuum Society } } This Conference will be a satellite of the Annual Brazilian Vacuum Conference, to be held in Sao Carlos City - 24-27 July, 1994 } } Guaruja is a pleasant island-city on the South Atlantic Sea. The Conference will be held in a Hotel on the beach. Proceedings will be refereed and published by an International Journal. } } For further information or abstracts submission contact : } } Prof.V.Baranauskas } Faculdade de Engenharia Eletrica } Universidade Estadual de Campinas UNICAMP } 13083-970 - Campinas - SP - Brasil } } FAX: +55-192-391395 } e.mail vitor-at-dsif.fee.unicamp.br } } ============================================================================= } ----- End Message ----- } } } } ----- End Included Message ----- } } }
Via: uk.ac.birmingham.computer-centre.ibm3090; Fri, 28 Jan 1994 11:37:44 +0000
Images from Nanoscope III can be saved as TIF files and studied on a PC using the VISILOG Software. This software is distributed (english language) by NOESIS Vision Inc 6800 Cote de Liesse Suite 200 Ville St Laurent, Quebec, Canada H4T 2A7 Tel (514) 345-1400 Fax (514) 345-1575
Fellow Microscopists: Here in the SEM lab, we scan a wide range of specimens (Insects, rodent teeth & skulls, fossil vertebrates & invertebrates) This is just to name afew. We use gold-palladium coating on these specimens. Is there anyone out there who has a sure fire method to remove coatings without damage to the specimen?
Thanks in advance, Peling Fong Melville
-------------------------------------------------------------- Peling Melville peling-at-amnh.org Interdepartmental Laboratories American Museum of Natural History
Does anyone out there have a number for Sony for color printer tech support? The loony I got in the DC area told me that I couldn't do half of the things that were printed right in the manual.
Jay: I have always made my EM slides using EM film and the only time I use glass is when I am making a composite slide. EM film mounts quite nicely in cardboard mounts, if your not doing any taping of the slides as you might do in making composite slides, why use glass? I have alwas had problems when I had to use Gepe glass. The best glass to use is by EMDE. They make an ultra thin glass (cat.# 135-NRT) slide making kit for the prevention of Newton Rings. Normal thickness glass can kill you every time with EM film. Don't know if EMDE is still in business. My box says Los Angeles 25, California. It's a few years old. You can call information in Los Angeles and see if they still have a phone # or check one of the big industrial photo supply houses or photo shop. Good Luck! Phil Rutledge prutle1-at-gl.umbc.edu
Further to Ron Anderson's suggestion: Bernie Kestel has written a paper "Non-Acid Electrolyte Thins Many Materials for TEM Without Causing Hydride Formation" (Ultramicroscopy 19 (1986) pp 205-212. I have a copy of the paper and would be pleased to FAX it to you and/or mail you a copy. You may contact me via CompuServe or as follows:
David Henriks South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 USA
TEL: 714-492-2600 FAX: 714-492-1499
You may also reach Bernie Kestel at: TEL: 708-252-4945 FAX: 708-252-4798
I also have a bibliography of over 40 papers dealing primarily with TEM sample preparation. I would be pleased to send you a copy and can subsequently provide reprints at no charge.
Bernie Kestel, Argonne National Laboratory's expert at electropolishing, suggests the following for Ti alloys:
13% HCl and 87% methanol temperature = -50 C 90 Volts and 35 mA in a single jet electropolishing machine
or
60ml Percholoric Acid 590 ml Methanol 350 ml 2-butoxy ethanol (butyl cellosolve) temperature = 0 C 40-50 Volts and 50 mA in a single jet electropolishing machine
or
5.30 g LiCl 11.16 g Mg(ClO4)2 500 ml methanol 100 ml 2-butoxy ethanol (butyl cellosolve) temperature = -40 or -50 C 150 to 250 volts and 30 mA flow speed = medium single jet electropolishing machine (South Bay) The flow restriction of the Tenupol holder may require less of the 2-butoxy ethanol which increases the viscosity of the solution. Results seem less satisfactory as the temperature is raised. You may need another power supply for the higher voltage. Greater or lesser amounts may be tried of the LiCl and Mg(ClO4)2. Add the powders to the methanol with simultaneous stirring. Do not reverse the voltage polarity: this introduces hydrogen into the specimen. Keep the specimen at a positive voltage. This approach worked well on compacted Ti nanocrystalline wafers.
Message-Id: {9401282054.AA22451-at-riker.ml.wpafb.af.mil} To: WALCKSD:ml:wpafb, SCHELTFJ:ml:wpafb Subj: Electropolishing of Titanium Orig-Author: {MACLAREI-at-ibm3090.computer-centre.birmingham.ac.uk}:ddn:wpafb ----------------------------------------------------------- Dear all,
Thank you to the two of you (Rob Tayloe and Cameron Begg) who replied to my earlier message concerning the electropolishing of Titanium for TEM specimens. However I can't believe that there is no one out there that has personal experience of preparing specimens of Ti or dilute alpha-Ti alloys for the TEM. If there is anyone out there who has done some TEM on such materials I would be very grateful if you could describe the specimen preparation route that you used.
Message-Id: {9401281628.AA21291-at-riker.ml.wpafb.af.mil} To: WALCKSD:ml:wpafb, SCHELTFJ:ml:wpafb Subj: Electropolishing of Titanium Orig-Author: {MACLAREI-at-ibm3090.computer-centre.birmingham.ac.uk}:ddn:wpafb ----------------------------------------------------------- Dear all,
Thank you to the two of you (Rob Tayloe and Cameron Begg) who replied to my earlier message concerning the electropolishing of Titanium for TEM specimens. However I can't believe that there is no one out there that has personal experience of preparing specimens of Ti or dilute alpha-Ti alloys for the TEM. If there is anyone out there who has done some TEM on such materials I would be very grateful if you could describe the specimen preparation route that you used.
Message-Id: {MAILQUEUE-101.940128134052.576-at-parmly1.parmly.luc.edu} To: microscopy-at-anlemc.msd.anl.gov
} Fellow Microscopists: } Here in the SEM lab, we scan a wide range of specimens (Insects, } rodent teeth & skulls, fossil vertebrates & invertebrates) This is just to } name afew. We use gold-palladium coating on these specimens. Is there } anyone out there who has a sure fire method to remove coatings without } damage to the specimen? } } Thanks in advance, } Peling Fong Melville } } -------------------------------------------------------------- } Peling Melville peling-at-amnh.org } Interdepartmental Laboratories } American Museum of Natural History
I have never heard of such a method, and do not believe that one exists. If I'm wrong, I would be VERY interested in hearing of it. But why bother? A light coat of Au/Pd won't hurt specimens, and coated SEM specimens can be embedded for TEM or LM. If there is some compelling reason not to have a coat on your specimen, the only choice is to examine them uncoated at low kV. This is pretty simple for bones & teeth, and many inverts. It is also doable with arthropods, even allowing for their setae, but you may need to go down to 300-500 V accelerrating voltage. Phil Oshel po-at-parmly1.parmly.luc.edu
Peling: Trying to remove gold-palladium coatings can be done on specimens such as yours but it can be a pain. I have used 20-50% acetone in water and sonicated the specimens. On some specimens this has worked and on others it hasn't. I know of no sure fire method to do this. What you can use instead of gold-palladium, is silver. I have removed silver off of hard and soft tissue samples rather easily. Sometimes I need to look at cells or whole tissue by SEM and then using the same sample for TEM. I use silver most (95%) of the time. At high mags in the SEM silver has a much smaller grain size than gold or gold-palladium and I get a better signal. All it takes to remove the silver is a basic darkroom chemical called Farmers Reducing Agent. This can be bought at just about any photo shop. You just soak your sample in this solution (full strength) until the silver is removed, then do what you need to do to the sample. If I am processing for TEM, I wash the specimen 3 x 15 minutes in distilled water first, 3 x 15 minutes in buffer and start my normal processing routine for TEM starting with the dehydration cycle. If you have any questions call me or Email me. Phil Rutledge, Director Center for Electron Microscopy UMBC Bio. Dept. Phone: (410) 455-3852 FAX: (410) 455-3875 Email: prutle1-at-gl.umbc.edu
Greetings, I am looking for a postdoc to join my lab to work on the relationship between cellulose synthesis and morphogenesis. Experience with either polarized light or electron microscopy, or with spatial measurements of growth is desirable. I encourage persons of all sexes, sexual orientations, colors, nationalities, cultures, sizes, shapes and physicalities to apply. Please send me your cv, names and contact #'s (or email handles) of several references, and if available, a statement of your "world view". If you have further q's, please ask. A brief description of the project, as funded, follows.
What is the role of cellulose deposition in controlling the degree of growth anisotropy? Although it is well known that highly anisotropic growth can be rendered isotropic by the randomization of the direction of deposition of cellulose microfibrils, it is not known whether the degree of growth anisotropy is also governed by the alignment of cellulose microfibrils. The major approach taken in this project is to measure growth anisotropy at a cellular scale and then quantify the alignment of cellulose microfibrils. Cells growing at various degrees of growth anisotropy will be obtained both by taking advantage of natural variations and through use of low concentrations of microtubule inhibitors. Cellulose alignment will be quantified throughout the whole wall with polarized light microscopy and at the innermost layer in metal-carbon replicas viewed in em. Experimental material will be roots of Arabidopsis and elongating tissue culture cells. There is support to work with Prof Andrew Staehelin at Boulder to use rapid-freezing deep etch techniques to make replicas without previous fixation. The project will also focus on several root morphology mutants in arabidopsis. These appear to have tissue-specific defects in cellulose deposition, and the relation between their processing of cellulose and their aberrant morphology will be studied.
Thanks for your interest,
Tobias Baskin
******************************** *************** Tobias I. Baskin 109 Tucker Hall /~~~\ Biol. Sci's * Univ. of Missouri c|o o\ Columbia, MO 65211 USA \ = / Tel:314-882-0173 FAX 314 - 882 - 0123 """ email: baskin-at-biosci.mbp.missouri.edu
Years ago we used to study cultured hepatocytes with SEM. These cell were coated with gold/palladium. Following SEM studies we would embed these specimens in epon and section the same cells for TEM. This worked beautifully and gave the cells an interesting electron dense outline. Our SEM fix, of course, was a good glutaraldehyde/osmium fix so that the TEM fine stucture was good.
Greetings, Does anyone have any comments on the new(ish) 35 degree diamond knives that are sold by Diatome (and others?)? I am interested in comments about "ambient temperature" knives, not "cryo" knives. The Tech Staff at Diatome say that the 35 degree knives reduce compression in some samples but not all. They further said that there were NO disadvantages to the 35 degree knives. My application is with growing plant tissues, such as roots.
I am a newcomer to electron microscopy and I appreciate help from the many wise folks who are a part of this list.
Thanks in advance, Tobias Baskin
******************************** *************** Tobias I. Baskin 109 Tucker Hall /~~~\ Biol. Sci's * Univ. of Missouri c|o o\ Columbia, MO 65211 USA \ = / Tel:314-882-0173 FAX 314 - 882 - 0123 """ email: baskin-at-biosci.mbp.missouri.edu
I am investigating methods of laser illumination for light microscopy. Specifically, I would like recommendations on methods of get uniform laser illumination free of laser speckle and noise. I am aware of one reference,
Ellis, G. W., "A fiber-optic phase-randomizer for microscope illumination by laser" _J. Cell. Bio._, vol 83, p303a.
but it is rather old, 1979, and I wonder if there are better ways of doing it nowadays. It was suggested to me that a phase randomized fiber bundle might do the trick. I was wondering if anyone has tried this? Although the spatial phases of the light will then vary across the fiber bundle area, I wonder if you will still get a well defined interference pattern, which will require vibration to average away and obtain uniform illumination?
Any ideas, experiences, or references will be appreciated. And I will summarize to the list the information I receive.
Thanks, eer
Erik Reuter, e-reuter-at-uiuc.edu, 217-244-6378(W), 359-4683(H), 244-6375 fax
We are in the market to purchase some microscope objectives to operate in the near infra-red, wavelength range of 700nm to 1100nm, to be used in a lab-built microscope. We would like to achieve the highest resolution allowed by diffraction at these wavelengths (~ 1 micron?) with dry objectives.
The microscope consists of a coaxial laser illuminator, a dichroic beam splitter, an objective lens, and a eyepiece/C-mount adapter/video camera. We are examining III/V semiconductor samples.
I would like to hear your experiences and recommendations on suppliers or manufacturers of infra-red microscope objectives. Are reflecting objectives our best bet? Who makes the best reflecting objectives? In order to obtain the best performance, should we purchase the objectives together with eyepiece/C-mount adapter or camera adapters, since the objectives are probably made for a specific correction?
Thanks, eer
Erik Reuter, e-reuter-at-uiuc.edu, 217-244-6378(W), 359-4683(H), 244-6375 fax
You can get excellent illumination by using a single-mode, polarization preserving fiber coupled to an output beam expander and collimator to achieve the desired beam size for input into a CLSM for example (6 mm). BY using an adjustable collimator the gaussian beam shape is preserved at any size beam and can be adjusted to the proper back focal plane size of your objective.
All multi-mode fibers will give speckles and you have to do something to scramble this at the output (vibration, optical scramblers). See the literature on fiber optics to see why this is always going to happen to your original gaussian beam.
We have a nice fiber and input coupler from Point Source in Winchester, England FAX 44-703-602470 and are using a Spindler & Hoyer output coupler, Goettingen, FRG FAX 49-551-69-35-166.
On Mon, 31 Jan 1994 16:49:17 +0200, { JOHNA-at-SCI.WFEB.EDU} wrote:
} } Years ago we used to study cultured hepatocytes with SEM. These cell were } coated with gold/palladium. Following SEM studies we would embed these } specimens in epon and section the same cells for TEM. This worked } beautifully and gave the cells an interesting electron dense outline. Our } SEM fix, of course, was a good glutaraldehyde/osmium fix so that the TEM } fine stucture was good. } } John Aghajanian.........JOHNA-at-sci.wfeb.edu
I do not understand why people want to get rid of the coating. It does not harm the TEM-specimen preparation. It does not harm the images. So what's the big need for removal. Actually, if I see a report stating that the same specimen has been first examined in a SEM with a metal coating and then prepared for TEM and there is no evidence of metal coating, bells would ring in my head and I would doubt wether the same samples are in question or not.
/jm
- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - Jouko K. Maki Navigare necesse est... Laboratory Manager, Ph.D. Laboratory of Electron Microscopy Kiinamyllynkatu 10 FIN-20520 TURKU FINLAND University of Turku Tel.: + 358 21 633 7318 INTERNET: jouko.maki-at-utu.fi FAX.: + 358 21 633 7380
ABSTRACTS FOR THE JOURNAL OF MICROSCOPY - FEBRUARY 1994
Journal of Microscopy, Vol. 173, Pt. 2, February 1994, pp. 87--101.
ESTIMATION OF THE DIRECTIONAL DISTRIBUTION OF SPATIAL FIBRE PROCESSES USING STEREOLOGY AND CONFOCAL SCANNING LASER MICROSCOPY
T. MATTFELDT,* A. CLARKE"" & G. ARCHENHOLD"", *Department of Pathology, University of Ulm, Oberer Eselsberg M23, D-89081 Ulm, Germany. ""Molecular Physics and Instrumentation Group, Department of Physics, University of Leeds, Leeds LS2 9JT, U.K.
Fibrous structures like polymers, glass fibres, muscle fibres and capillaries are important components of materials and tissues. A spatial fibre process is the union of smoothly curved or linear one-dimensional features of finite length, arranged in an unbounded three-dimensional reference space according to some random mechanism. Design-based stereology was combined with confocal scanning laser microscopy to study samples of fibre-reinforced composites, which were considered as realizations of not necessarily isotropic fibre processes. The methods enable the unbiased estimation of the intensity and of the directional distribution of spatial fibre processes from arbitrarily directed pairs of registered parallel optical sections a known distance apart. The directions of fibres sampled by a frame of observation on the reference plane are estimated from the coordinates of the intersection points of the fibres with both optical planes using confocal scanning laser microscopy. The true directional distribution of the fibre process is estimated by weighting each measured direction by the reciprocal of its chance of being sampled, which can be inferred from the data. The concept of complete directional randomness for uniformly and independently distributed spatial directions is introduced. The cumulative distribution function of the angular distances between different directions and other exact relations are derived for complete randomness of vectorial and axial directions. A Monte Carlo method is constructed to test spatial fibre processes, whose fibres have negligibly small curvature, for complete directional randomness. Confocal scanning laser microscopy was used to study the angular distribution of glass fibres in a polymer composite which was subjected to increasing hydrostatic extrusion. The hypothesis of complete directional randomness had to be rejected for all samples with 1% probability of error. The directional distribution was of the bipolar type, with the principal axis directed parallel to the axis of extrusion. Progressive stretching of the material increased the degree of anisotropy of the glass fibres. Although presented for an application in polymer physics, the methods are general and may also be applied in biological investigations.
Journal of Microscopy, Vol. 173, Pt. 2, February 1994, pp. 103--114.
AUTOMATED TRACING AND VOLUME MEASUREMENTS OF NEURONS FROM 3-D CONFOCAL FLUORESCENCE MICROSCOPY DATA
A. R. COHEN,* B. ROYSAM* & J. N. TURNER*"", *ECSE Department, Rensselaer Polytechnic Institute, Troy, NY 12180-3590, U.S.A. "" Wadsworth Center for Laboratories and Research, New York State Department of Health, Albany, NY 12201-0509, U.S.A.
Three-dimensional (3-D) image analysis algorithms and experimental results that demonstrate the feasibility of fully automated tracing of neurons from fluorescence confocal microscopy data are presented. The input to the automated analysis is a set of successive optical slices that have been acquired using a confocal scanning laser microscope. The output of the system is a labelled graph representation of the neuronal topology that is spatially aligned with the 3-D image data. A variety of topological and metric analyses can be carried out using this representation. For instance, precise measurements of volumes, lengths, diameters and tortuosities can be made over specific portions of the neuron that are specified in terms of the graph representation. The effectiveness of the method is demonstrated for a set of sample fields featuring selectively stained neurons. Additional work will be needed to refine the method for unsupervised use with complex data involving multiple intertwined neurons and extremely fine dendritic structures.
Journal of Microscopy, Vol. 173, Pt. 2, February 1994, pp. 115--126.
ALGORITHMS FOR AUTOMATED CHARACTERIZATION OF CELL POPULATIONS IN THICK SPECIMENS FROM 3-D CONFOCAL FLUORESCENCE MICROSCOPY DATA
B. ROYSAM,* H. ANCIN,* A. K. BHATTACHARJYA,* M. A. CHISTI,"" R. SEEGAL"" & J. N. TURNER"", *Rensselaer Polytechnic Institute, Troy, NY 12180-3590, U.S.A. "" Wadsworth Center for Laboratories and Research, New York State Department of Health, Albany, NY 12201-0509, U.S.A.
Methods are presented for the automated, quantitative and three-dimensional (3-D) analysis of cell populations in thick, essentially intact tissue sections while maintaining intercell spatial relationships. This analysis replaces current manual methods which are tedious and subjective. the thick sample is imaged in three dimensions using a confocal scanning laser microscope. The stack of optical slices is processed by a 3-D segmentation algorithm that separates touching and overlapping structures using localization constraints. Adaptive data reduction is used to achieve computational efficiency. A hierarchical cluster analysis algorithm is used automatically to characterize the cell population by a variety of cell features. It allows automatic detection and characterization of patterns such as the 3-D spatial clustering of cells, and the relative distributions of cells of various sizes. It also permits the detection of structures that are much smaller, larger, brighter, darker, or differently shaped than the rest of the population. The overall method is demonstrated for a set of rat brain tissue sections that were labelled for tyrosine hydroxylase using fluorescein-conjugated antibodies. The automated system was verified by comparison with computer-assisted manual counts from the same image fields.
Journal of Microscopy, Vol. 173, Pt. 2, February 1994, pp. 127--141.
MEASUREMENT OF ABSOLUTE TRACER CONCENTRATIONS IN TISSUE SECTIONS BY USING DIGITAL IMAGING FLUORESCENCE MICROSCOPY. APPLICATION TO THE STUDY OF PLASMA PROTEIN UPTAKE BY THE ARTERIAL WALL
P. D. Weinberg,* C. P. Winlove"" & K. H. Parker"", *Department of Biochemistry and Physiology, University of Reading, Whiteknights, PO Box 228, Reading RG6 2AJ, U.K. "" Centre for Biological and Medical Systems, Imperial College of Science, Technology and Medicine, Prince Consort Road, London SW7 2BY, U.K.
Digital imaging fluorescence microscopy (DIFM) of tissue sections was used to quantify uptake of labelled plasma proteins by the arterial wall. Several aspects of the measuring system were investigated so that absolute tracer concentrations and their local variation could be derived from digitized images. These investigations may be relevant to other studies employing DIFM. Nonlinearities were found to arise from offsets in the video digitizers, from background fluorescence and stray light within the microscope and from the transfer characteristics of the intensified CCD camera. Camera gain controls showed complex behaviour. Camera output fell substantially for several hours after switching on and was affected by room temperature. Large spatial variations in response were caused by the geometry of the microscope optics and by the image intensifier. However, the ratios between areas were not affected by light intensity or camera gain settings. Measured intensities were independent of the depthwise location of fluorophores within tissue sections but they were affected by the emission from objects outside the measuring area. Photobleaching of tracer varied significantly over the range of excitation intensities and durations used but was not concentration dependent. Methods used to correct these effects and obtain a uniform, linear and constant relationship between concentration and grey level are described. Using the system and appropriate corrections, in vivo uptake of sulphorhodamine-B-labelled serum albumin by the rabbit aortic wall was investigated. Results obtained for the mean uptake of tracer and its local variation were quantitatively similar to those previously obtained with nonmicroscopic methods.
Journal of Microscopy, Vol. 173, Pt. 2, February 1994, pp. 143--147.
A CRYOGLUE TO MOUNT VITREOUS BIOLOGICAL SPECIMENS FOR CRYOULTRAMICROTOMY
K. RICHTER, Laboratoire d'Analyse Ultrastructurale, CH-1015 Lausanne, Switzerland
Mixtures of ethanol, 2-propanol and 2-butanol can be used as a cryoglue to mount vitrified biological specimens for ultrathin cryosectioning. Brought directly from room temperature to a cutting support at 140 K in the cold chamber of a cryoultramicrotome, these alcohols stiffen to a viscous and gluey consistency allowing the attachment or embedding of a vitreous biological sample. The mass hardens at lower temperatures fixing the sample well for microtomy. With ethanol: 2-propanol (2:3), samples are applied at 140 K and ultrathin cutting can be done at 115 K.
Journal of Microscopy, Vol. 173, Pt. 2, February 1994, pp. 149--154.
THE USE OF THE STABLE ISOTOPE 44Ca IN STUDIES OF CALCIUM INCORPORATION INTO DENTIN
T. LUNDGREN, E. U. ENGSTROM,* R. LEVI-SETTI+, A. LINDE & J. G. NOREN++, Departments of Oral Biochemistry and ++ Pedodontics, Faculty of Odontology, University of Goteborg, Sweden. * Department of Physics, Chalmers University of Technology, Sweden. + Enrico Fermi Institute and Department of Physics, University of Chicago, U.S.A.
The incorporation into rat incisor dentin of two calcium isotopes, the stable 44Ca and the radioactive 45Ca, was studied using secondary ion mass spectrometry (SIMS) step-scanning and imaging, and autoradiography, respectively. The results demonstrated a time-dependent incorporation of the calcium isotopes into the mineral phase of dentin. With the SIMS step-scanning, detecting 44Ca, the ion yield was high in the odontoblasts 2 min after intravenous injection. After 10 min a marked increase in signal intensity was found at the dentin mineralization front. This result was consistent with those obtained by 45Ca autoradiography; a peak of incorporation occurred 10 min after injection of the isotope. Likewise, localization of 44Ca to the mineralization front could be demonstrated 10 min after injection by SIMS imaging. In images obtained at earlier intervals, no such increase in ion yield could be detected. The results show that the nonradioactive, stable isotope 44Ca can be used as a marker for biomineralization in a similar way to radioactive 45Ca.
Journal of Microscopy, Vol. 173, Pt. 2, February 1994, pp. 155--164.
STRUCTURAL FEATURES OF CELL WALLS FROM TOMATO CELLS ADAPTED TO GROW ON THE HERBICIDE 2,6-DICHLOROBENZONITRILE
B. WELLS,* M. C. McCANN,* E. SHEDLETZKY,"" D. DELMER"" & K. ROBERTS*, * Department of Cell Biology, John Innes Institute, Colney Lane, Norwich NR4 7UH, U.K. "" Department of Botany, Institute of Life Sciences, The Hebrew University, Jerusalem 91904, Israel
Evidence from high-resolution images of primary cell walls suggests that the cell wall is constructed from at least two independent yet coextensive fibrous networks, one based on cellulose/hemicellulose and the other on pectin. The ability to analyse the structure of each of these networks in isolation has been hampered by a lack of suitable biological material such as mutants. However, the recent use of the cellulose-synthesis inhibitor 2,6-dichlorobenzonitrile (DCB) that prevents the formation of the cellulose--xyloglucan network while allowing the pectin network to form a functional wall offers the unique opportunity of studying at least the pectin network independently. A range of electron microscopy techniques and a novel spectroscopy method are used to study the walls from tomato suspension cells adapted to growth on DCB. Measurements of the minimum cell wall thickness derived from thin sections of dehydrated walls show that the marked reduction in level of the cellulose/hemicellulose network affects neither the thickness of the wall formed, nor the apparent spacing of pectin molecules. However, images obtained by the fast-freeze, deep-etch, rotary-shadowed (FDR) replica technique show that the three-dimensional architecture of these pectin-rich walls is very different from that of nonadapted walls. Fourier transform infrared (FTIR) microspectroscopy data and immunogold-labelling studies provide additional evidence that supports the previous biochemical data.
Journal of Microscopy, Vol. 173, Pt. 2, February 1994, pp. 165--172.
A PARTICLE SEGMENTATION METHOD BASED ON NUCLEATION AND GROWTH OF THE BACKGROUND
XIN ZHENG & MAX KOLB"", Laboratoire de Chimie Theorique, Ecole Normale Superieure de Lyon, 46, Allee d'Italie, 69364 Lyon Cedex 07, France. Institut de Recherches sur la Catalyse, CNRS, 2, Ave. A. Einstein, 69626 Villeurbanne Cedex, France
A new algorithm for image segmentation is proposed, which is capable of extracting particles in the presence of noise and background fluctuations. It begins with the detection of small regions belonging to the background, called background nuclei, and then lets these nuclei grow to become the entire background. Edge information and region information of the image are used simultaneously.
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Message-Id: {MAILQUEUE-101.940201080121.320-at-parmly1.parmly.luc.edu} To: microscopy-at-anlemc.msd.anl.gov
I second Jouko Maki's opinion below. I've never noticed that the metal coating on the specimen causes any damage (or other problems) to the knife in sectioning, & it can be cut with a glass knife. So, the presence of the coating could be considered required to demonstrate that the section was taken from a SEM specimen. Phil Oshel
} From: "Jouko M{ki" {jokamaki-at-utu.fi} } Subject: Re: removing SEM specimen coatings ... } } Years ago we used to study cultured hepatocytes with SEM. These cell were } } coated with gold/palladium. Following SEM studies we would embed these } } specimens in epon and section the same cells for TEM. This worked } } beautifully and gave the cells an interesting electron dense outline. Our } } SEM fix, of course, was a good glutaraldehyde/osmium fix so that the TEM } } fine stucture was good. } } } } John Aghajanian.........JOHNA-at-sci.wfeb.edu ... } I do not understand why people want to get rid of the coating. It does not } harm the TEM-specimen preparation. It does not harm the images. So what's } the big need for removal. } Actually, if I see a report stating that the same specimen has been first } examined in a SEM with a metal coating and then prepared for TEM and there } is no evidence of metal coating, bells would ring in my head and I would } doubt wether the same samples are in question or not. } } /jm } } - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - } Jouko K. Maki Navigare necesse est... } Laboratory Manager, Ph.D. } Laboratory of Electron Microscopy Kiinamyllynkatu 10 FIN-20520 TURKU FINLAND } University of Turku Tel.: + 358 21 633 7318 } INTERNET: jouko.maki-at-utu.fi FAX.: + 358 21 633 7380 }
In my efforts to produce even better sections of these 35 um glass beads, I spoke with the folks at Diatome last week. They advised me to try a new 45 degree diamond knife (I had asked her about a 55 degree knife as someone had suggested on this bulletin board) and then to try a 35 degree knife. They predicted that we would get our best sections with the 35 knife. Sectioning glass beads with a 35 knife, of course, would cause Gib Ahlstrand to cringe. But we're going to try it to see how it goes. I'll keep you abreast of our progress. The prediction is: good sections and a very short edge life.
Message-Id: {9402011548.AA10683-at-umail.UMD.EDU} To: microscopy-at-anlemc.msd.anl.gov
************************************************************************ The University of Maryland Short Course
PRACTICAL ASPECTS OF SCANNING ELECTRON MICROSCOPY
Session I: March 14 - March 18, 1994
Session II : March 21 - March 25, 1994
This intensive four an one-half day course provides participants with a thorough coverage of the basic theory and practice of scanning electron microscopy and related techniques, including X-ray microanalysis. The course employs easy to understand lectures coupled with supervised laboratory exercises to provide practical instruction in the operation of scanning electron microscopes equipped with modern accessories. Class size is limited assuring those attending of the opportunity for extensive hands on experience on the instrumentation available. Attendees in the past have represented a broad range of disciplines (materials, semiconductors, failure analysis, pharmaceuticals, food science, forensics, biology, etc.).
For more information about the course, contact: Tim Maugel University of Maryland Department of Zoology College Park, MD 20742 Phone : (301)405-6898 Fax : (301)314-9358 E-Mail : maugel-at-zool.umd.edu *************************************************************************** Tim Maugel University of Maryland Laboratory for Biological Ultrastructure Department of Zoology College Park, MD 20742 Phone: (301)405-6898 Fax: (301)314-9358 EMail: tm11-at-umail.umd.edu
We are using Unicryl as embedding medium for immunoEM experiments. One of the problems is that you cannot use Os before embedding. Does anyone have experience with on grid Os after immuno. Which grids? Which protocol?
We are at the moment trying to use a Vidas system from Kontron to do some ploidy analysis using Feulgen. We have to calibrate the system using well known 2n, 3n, 4n , .... cells. Does anyone have an idea were to get such celllines or slides for calibration.
Thanks to everyone for the many useful comments on scrubbers used for RIE etching. There seems to be a consensus that some sort of scrubbers are required before exhausting into atmosphere. I have checked some of the chemical supply house and point of use scrub- bers are available for modest cost as suggested. If a scrubber is not already attached to main exhaust in the laborstory, the point of use scrubber would be more than adequate for low or moderate users of the RIE system.
Chlorine gas for etching aluminium presents a more costly problem due to its toxicity and the safety requirements for using in an enclosed laboratory. Does anyone have experience/recommendation for alternative gases for the aluminium etch that would not have toxicity requirments for handling and operation and as a conse- quence be less costly to handle/use.
Thanks again.
Richard Sartore US Army Research LAboratory AMSRL-EP-RA Fort Monmouth, NJ 07703 RSARTORE-at-MONMOUTH-ETDL1.ARMY.MIL
John Mansfield North Campus Electron Microbeam Analysis Lab University of Michigan 2455 Hayward Ann Arbor MI 48109-2143 Phone: (313)936-3352 FAX (313)936-3352 jfmjfm-at-engin.umich.edu or John.F.Mansfield-at-umich.edu Time: 4:49 PM Date:2/2/94 NC EMAL
Applications of Environmental Scanning Electron Microscopy
Dear Fellow ESEM, Wet SEM & Low Vacuum SEM User(s):
Many of the electron microscope manufacturers now offer an "environmental", "low-vacuum" or "wet" SEM as part of their product lines. These instruments have the unique capability of forming secondary or back-scattered electron images while the sample is in what would typically be considered an extremely poor vacuum. While novel, these instruments have not yet been taken too seriously by the microscopy community.
To highlight the flexibility and functionality of these low-vacuum systems, this year's joint Microscopy Society of America and Microbeam Analysis Society meeting in New Orleans will feature a symposium sponsored by MAS entitled "Applications of Environmental or Low-Vacuum Scanning Electron Microscopy".
This is an invitation to you and your colleagues to contribute papers to this symposium.
This symposium will be broadly based and not material, instrument or technique specific. Submissions covering novel in-situ experiments; advances in hot-stage, cold-stage or deformation microscopies and studies on non-conducting materials, ceramics, polymers etc., liquids and emulsions are encouraged. More "conventional" SEM experiments performed in an environmental SEM are also appropriate
Organizers: Roger Bolon General Electric Corporate R&D Building K-1 Schenectady NY 12309 Tel: (518) 387-7783 email: bolonrb-at-crd.ge.com
Stuart McKernan University of Minnesota High-Resolution Microscopy Center 100 Union St. S.E. Minneapolis MN 55455-0153 Tel.: (612) 624-6590 e-mail: stuartm-at-maroon.tc.umn.edu
John Mansfield University of Michigan North Campus EMAL 413 SRB 2455 Hayward Ann Arbor MI 48109-2143 Tel: (313) 936-3552 email: John.F.Mansfield-at-umich.edu
Abstract deadline is 15th March Please submit TWO PAGE manuscripts (plus three copies) to: Meeting Office, PO Box EM, Woods Hole MA 02543.
} From: lherault-at-acs.bu.edu (Ronald LHerault) } Date: Wed, 2 Feb 94 12:33:32 -0500 } To: microscopy-at-anlemc.msd.anl.gov } Subject: Removing coatings.
} We often use replication on specimens that can't be changed by coating. } Since this is a dental school, we have ready access to polyvynil } oops, polyvinyl siloxane impression materials. The negatives produced } are filled with Spurr low viscosity embedding material and oven cured. } We then mount and coat the specimen for SEM. The only time I ever } had a problem was when I took an impression of a sand dollar. The } impression material stuck in the many pores o the saample.
I am not familiar with this material, other than having it used on me--is it anything like silicone bathtub chaulk? I've used that to fill the lateral line canals of fish, and have gotten very nice impressions of the sensory neuromasts, sensory strip and all (including possible pits of hair cells!?) Phil Oshel please see c.v. on MSA bulletin board--job cut
A few spaces remain in the upcoming school on EELS imaging and analysis techniques to be held at the Gatan R&D facility in Pleasanton, California during 1-4 March 1994. The school covers the basic theory and practice of electron energy loss spectroscopy and energy filtered imaging in transmission electron microscopy and provides opportunities for hands-on experience with these techniques, both in data acquisition directly at the microscope and in data analysis at computer work stations. Registration is open to all interested parties. The school is run at cost, the registration fee of US $750 covering tuition, course materials, and lunches (travel, hotel, and other meals are not included). For further information, please contact
Mike Kundmann Gatan EELS Software R&D 6040 Washington Street Downers Grove, IL 60516-1978 USA (708) 964-2153 (24-hour voice message and/or fax)
Spaces remain in the upcoming school on Digital Microscopy to be held at the Gatan R&D facility in Pleasanton, California during 22-25 February 1994. The school covers digital image capture and analysis techniques and their application to computer-assisted operation of scanning and (fixed-beam) transmission electron microscopes. The school also provides opportunities for hands-on experience with these techniques, both in image capture directly at the SEM or TEM and in data analysis at computer work stations. Registration is open to all interested parties. The school is run at cost, the registration fee of US $750 covering tuition, course materials, and lunches (travel, hotel, and other meals are not included). For further information, please contact
Jacob Wilbrink Gatan R&D 6678 Owens Drive Pleasanton, CA 94588-3334 USA Phone: (510) 224-7316 (24-hour voice mail) Fax: (510) 463-0204
I'm interested in the morphology of Panthera Pardus epidermis. Does anyone know if there is something unusual about the epithelial cells of this animal skin?
============================================================================= Francisco Javier Hernandez Blazquez | Universidade de Sao Paulo - Faculdade de | e-mail fjhblazq-at-fox.cce.usp.br Zootecnia e Engenharia de Alimentos | Voice: 55 195 616122 r. 265 Departamento de Ciencias Basicas/Histologia| Av. Duque de Caxias Norte, 225 CP 23 | Fax: 55 195 611689 CEP 13630-000 Pirassununga (Sao Paulo) | BRAZIL | ==============================================================================
We are attempting to set up some image processing capabilities on our SEM. Does anyone have any information on systems that can do image analysis (particle size, phases, etc) on a pc or Mac. Any comments on likes and dislikes would be extremely helpful, as would neighborhood prices.
} We are attempting to set up some image processing capabilities on our SEM. } Does anyone have any information on systems that can do image analysis } (particle size, phases, etc) on a pc or Mac. Any comments on likes and } dislikes would be extremely helpful, as would neighborhood prices. } } Thanks!!
A little more information would be helpful. Do you want to do image capture and then analysis of those images? Is your SEM analog or digital? Do you want EDS, too?
One excellent option to look at is the system from 4pi Analysis, Inc. Their system can interface with digital or analog scopes. They capture into NIH-Image, the public domain program from NIH that runs on the Mac. They can also add the capability to do EDS, but that's not required. Their prices are very reasonable. Their image capture board, which resides in a NuBus slot takes control of the beam for slow scanned images.
I saw one of their units in a busy, high volume, high quality lab that could buy pretty much what they want. They have the 4pi and tell me they're very pleased with it.
Dapple also has a system for image capture. Theirs is passive, and takes the image the scope gives it. It's also much more expensive.
4pi Analysis, Inc. 3500 Westgate Drive, Suite 403 Durham, NC 27707 (919) 489-1757
Dapple Systems, Inc. 355 W Olive Ave. #100 Sunnyvale, CA 94086 (408) 733-3283
John chandler-at-lamar.ColoState.EDU Fort Collins, CO
We've simply cabled a framegrabber board from a mac running NIH Image to the SEM video port. This means manually synching the grab to the scan sweep, but its cheap and effective. You could probably do the same with any PC based image system. I've read about the 4pi system and that would prbably be our choice when our needs for SEM digital capture expand. Glen MacDonald Hearing Development Laboratories RL-30 University of Washington Seattle, WA 98195 (206)543-8360 glenmac-at-u.washington.edu
I want to take the image from a ccd camera on my optical microscope, combine it with just a few parameters (temperature, sample positioning, etc) output from a pc and combine them for recoding on a video recorder. Anyone got any ideas?
I am not a subscriber to the list. Please post to me personally. If requested, I will summarize responses as best I can.
A friend is looking for a computer program written in C or Pascal (or any language compatible with the hardware) that would enable her to use the arrow keys on the PC keyboard to move a mechanical microscope stage in the x and y directions via a PC21 Indexer card and compumotor stepper motor. We are looking for program or possible source for program or better place to post this request. Thanks for your help.
Keith Hallam writes: } I just want to take the image from a ccd camera... a few parameters ...
Just thought I would offer you the following:
My company makes a series of small devices called video marking and measuring systems. They hook up in series with the video to your monitor and give you all kinds of capabilities. For example: Add labels to the image. You choose the size and style. Add markers. Choose among 16 styles, up to 16 different ones at once Use a stopwatch Count things by marking them. Tally shows on screen. Measure distances, areas, pathlengths, radius, lots of options. Draw anything at all freehand. Put up grids, targets, reference ruler. Other stuff that I can't think of right now. Oh.. Send data to a printer or pc.
This may not be what you were looking for, but these systems were designed for microscope use, and are being used by quite a few people at this time. They have only been out for about 6 months.
The problem, I think, is that you wanted to take the data from your pc automatically. This system can't do that, but it does offer a lot of things that you may find useful.
The marking and measurement system is called the XR-2000 series, and comes in a few flavors, depending on what type of video you have.
We also make a stand alone real time video processor called the OMNEX which does many of these things and also video averaging, integration, memory stuff, low light camera control, and all kinds of other things.
The XR-2000 marking and measurement systems cost around $1700 depending on the type of video. The OMNEX processor is about $5350.
Deadline for advance registration and submission of papers for the ICEM13 in Paris is March 1, 1994. Before March 1st the registration fee is 2000FFR and after March 1st it increases to 2200FFR. Student registration is 1100 FFR at all times, including at the meeting. Due to problems with the mail, the organizing committee will be reasonably flexible about accepting the "before 1st March" rate. If registrations are post marked by the 1st of March, they will be accepted at the advance registration rate.
As for abstracts, they can also be flexible. Everyone must send a copy of their contribution to the ICEM13 before March 1st, using either fax or email for the text. These can then be sent to the symposia chair for review. HOWEVER, the printed copies with complete illustrations must reach France before the contributions go to the printers. If the fully-illustrated printed contributions are not received in time, they may be accepted as talks or posters, but will not appear in the proceedings.
To receive registration information, etc. (the 3rd Circular) for the Paris ICEM13, contact Dr. Bernard Jouffrey directly by email at: jouffrey-at-mssmat.ecp.fr or by fax at: 33 1 41 13 14 30. Send your name, address, fax no., and email no.
Another question, totally unrelated to the last one. Has anyone any experience, or references, to the use of image analysis to differentiate between different types of blood cells in an optical microscopy image?
To: Anyone interested From: Cesar D. Fermin, Ph.D. Tulane Pathology.
life. Excellent condition. Water chiller included.
Available free, but:
1) you must pay for packing and shipment to your destination,
2) You must arrange for packing at time of dismantling,#000# sometimes in April.
3) Respond via e-mail, but acceptance must be arranged in writing or via fax to me.
Cesar D. Fermin, Ph.D. Tulane University Medical School Department of Pathology 1430 Tulane Ave/SL79 New Orleans, La 70112-2699 (504) 584-2521 Fax 587-7389
Via: uk.ac.birmingham.computer-centre.ibm3090; Tue, 8 Feb 1994 11:46:36 +0000
We are considering purcahsing a Quadra 840AV for image capture and manipulation of phase, DIC and fluorescence microscopy of biological material, including quick-time - in particular of Platyhelminths. I would appreciate contact or advice from anyone with experience with this machine for similar applications. I understand that the image grabber board is probably inadequate for such applications and would need to be replaced by a higher quality card. What remaining advantages are there (apart from the audio which we are not interested in) for the AV machine over an 800 with the addition of the appropriate card?
There has probably been considerable discussion on this issue in the past in this group and in the NIH Image group and I would also appreciate instructions on where to look for this in an FAQ site.
Dr N.A. Watson Department of Zoology University of New England Armidale, NSW, 2351 AUSTRALIA Fax: 067 711 869 Phone: 067 732181
Tom Gore from Universtiy of Victoria asked about the Argus 10 processor. My company (Imagen Inc) makes the main competitor to the Argus 10, the OMNEX. We consider it to be an improvement on the Argus 10 in many ways.
The OMNEX is easy to use. It uses a mouse and on-screen menus. There are no manual controls at all.
The OMNEX can do real time averaging, integration, memory operations (subtraction, addition, etc.), take a gradient and lots more. It has digital contrast enhancement with 20 storage macros and a custom lut that you can draw with the mouse. It has distance, area, path, event, radius measurements with four calibrations. It does psuedocolor and can store 4 images and cycle them automatically (like a slide show).
It can control long exposure cameras for low-light work. There is a built-in help menu system, although most people don't need it because the menus are pretty straightforward.
There are quite a few other things that it does that I can't think of at the moment, but if you like, I can drop a brochure in the mail (the REAL mail). We have distributors all over US, Canada, and are now getting into Japan and Europe.
We also have a marking and measuring system that is quite nice. I can send you that info also.
Hope this tells you maybe what you were looking for. Of course, my opinion is absolutely politically unreliable, but the device stands for itself. People seem to enjoy using it. (It even plays breakout.)
We used Cr2O3 standard for Cr2O3 in microprobe analyses. Some time ago standard was changed to metallic Cr. When analysing Cr2O3 with new metallic standard we noticed that it gives analyses which are 4 wt% below. It is obvious that the ghange to metallic Cr standard caused the problem. Why Cr2O3 and metallic Cr as standards give different values ? Peak positions are same in both standards. Best wishes,
Jussi Liipo Department of Geology University of Oulu SF-90570 Oulu FINLAND
I teach SEM to undergraduate Biology students and would like to provide them with a brief introduction to specimen preparation techniques used by material scientists. Does anyone know of a review article or basic text on the subject that would be useful for this purpose? Thanks for any references you can provide!!!!
Kenneth M. Bart Director, Electron Microscopy Facility Hamilton College Clinton, New York 13323 USA kbart-at-hamilton.edu (315) 859-4715
I have the following surplus equipment in my lab that I would like to sell:
1. LKB 8800 Ultramicrotome
2. EmScope CPD 750 Critical Point Drier
Both instruments are in good working order, but have not been used for a few years. If anyone is interested please contact me directly via e-mail , do not reply on this Microscopy listserver.
Kenneth M. Bart Director, Electron Microscopy Facility Hamilton College Clinton, New York 13323 USA kbart-at-hamilton.edu (315) 859-4715
I need to obtain citations and/or procedures for the use of rutheniun red in the staining of mucopolysaccharides in thin and semi-thin sections.*************************************** David Garrett "DGARRETT-at-GAB.UNT.EDU" University of North Texas Dept. Biological Sciences (817)565-3964 Fax (817)565-4136 ***************************************
JOURNAL OF MICROSCOPY, VOLUME 173 PART 3, MARCH 1994 **************************************************************
Journal of Microscopy, Vol. 173, Pt. 3, March 1994, pp. 173--181.
ATOMIC FORCE MICROSCOPY OF FREEZE-FRACTURE REPLICAS OF RAT ATRIAL TISSUE
by L. KORDYLEWSKI, D. SANER & R. LAL, University of Chicago, Department of Medicine/Cardiology, 5841 S. Maryland Ave., Chicago, IL 60637, U.S.A.
Summary
Atomic force microscopy (AFM) has provided three-dimensional (3-D) surface images of many biological specimens at molecular resolution. In the absence of spectroscopic capability for AFM, it is often difficult to distinguish individual components if the specimen contains a population of mixed structures such as in a cellular membrane. In an effort to understand the AFM images better, a correlative study between AFM and the well-established technique of transmission electron microscopy (TEM) was performed. Freeze-fractured replicas of adult rat atrial tissue were examined by both TEM and AFM. The same replicas were analysed and the same details were identified, which allowed a critical comparison of surface topography by both techniques. AFM images of large-scale subcellular structures (nuclei, mitochondria, granules) correlated well with TEM images. AFM images of smaller features and surface textures appeared somewhat different from the TEM images. This presumably reflects the difference in the surface sensitivity of AFM versus TEM, as well as the nature of images in AFM (3-D surface contour) and TEM (2-D projection). AFM images also provided new information about the replica itself. Unlike TEM, it was possible to examine both sides of the replica with AFM; the resolution on one side was significantly greater compared with the other side. It was also possible to obtain quantitative height information which is not readily available with TEM.
Journal of Microscopy, Vol. 173, Pt. 3, March 1994, pp. 183--197.
TIP ARTEFACTS IN SCANNING FORCE MICROSCOPY
by U. D. SCHWARZ,* H. HAEFKE, P. REIMANN & H.-J. GNTHERODT, Institute of Physics, University of Basel, Klingelbergstrasse 82, CH-4056 Basel, Switzerland
Summary
Since its invention in 1986, scanning force microscopy (SFM) has experienced great success as a characterization method for topography on small scales. In spite of the enormous potential of the method, it is limited by the quality of the tip used for probing the surface topography. Convolutions of non-ideal tip shapes with the real topography and tip bending, flexing and jumping effects produce artefacts in the resulting images. A brief description of the preparation and characteristics of the most commonly used SFM tips is given. A variety of different artefacts originating from tip properties is presented and illustrated with selected scanning force micrographs. Methods to minimize tip artefacts in SFM images are described.
Journal of Microscopy, Vol. 173, Pt. 3, March 1994, pp. 199--210.
CALIBRATION OF THE SCANNING (ATOMIC) FORCE MICROSCOPE WITH GOLD PARTICLES
by S. XU & M. F. ARNSDORF, Department of Medicine, The University of Chicago, Chicago, IL 60637, U.S.A.
Summary
Scanning force microscopy (SFM) holds great promise for biological research. Two major problems that have confronted imaging with the scanning force microscope have been the distortion of the image and overestimation in measurements of lateral size due to the varying geometry and characteristics of the scanning tip. In this study, spherical colloidal gold particles (10, 20 and 40nm in diameter) were used to determine (1) tip parameters (size, shape and semivertical angle); (2) the distortion of the image caused by the tip; and (3) the overestimation or broadening of lateral dimensions. These gold particles deviate little in size, are rigid and have a size similar to biological macromolecules. Images of the colloidal gold particles by SFM were compared with those obtained by electron microscopy (EM). The height of the gold particles as measured by SFM and EM was comparable and was little affected by the tip geometry. The measurements of the lateral dimensions of colloidal gold, however, showed substantial differences between SFM and EM in that SFM resulted in an overestimate of the lateral dimensions. Moreover, the distortion of images and broadening of lateral dimensions were specific to the SFM tip used. The calibration of the SFM tip with mica provided little clue as to the type of distortion and the amount of lateral broadening observed when the larger gold particles were scanned. The SFM image also depended on the orientation of the tip with respect to the specimen. Our results suggest that quantitative SFM imaging requires calibration to identify and account for both the distortions and the magnitude of lateral broadening caused by the cantilever tip. Calibration with gold particles is fast and nondestructive to the tip. The raw imaging data of the specimen can be corrected for the tip effect and true structural information can be derived. In summary, we present a simple and practical method for the calibration of the SFM tip using gold particles with a size in the range of biomacromolecules that allows: (1) selection of a cantilever tip that produces an image with minimal distortion; (2) quantitative determination of tip parameters; (3) reconstruction of the shape of the tip at different heights from the tip apex; (4) appreciation of the type of distortion that may be introduced by a specific tip and quantification of the overestimation of the lateral dimensions; and (5) calculation of the true structure of the specimen from the image data. The significance is that such calibration will permit quantitative and accurate imaging with SFM.
Journal of Microscopy, Vol. 173, Pt. 3, March 1994, pp. 211--218.
CONTRIBUTION OF ENERGY-FILTERING TEM TO THE DETECTION OF CALCIUM: APPLICATION TO MAST CELLS
by M. HOROYAN,* M. SOLER,* J. M. MARTIN,""A.-M. BENOLIEL,* M. FRATERNO,~~ M. PASSEREL,## E. KATCHBURIAN, P. BONGRAND* & C. FOA*, *INSERM U 387, Laboratoire d'Immunologie, Hpital Ste Marguerite, 13277 Marseille Cedex 9, France. ""Ecole Centrale de Lyon, Departement de Technologie des Surfaces, URA CNRS 855, 36 Avenue Guy de Collongue, BP 163, 69131 Ecully Cedex, France. ~~Service commun de Microscopie lectronique, Facult de Mdecine, Bd Jean Moulin, 13005 Marseille, France. ##Centre de Microscopie lectronique et de Microanalyse, Facult des Sciences et Techniques de St Jrome, avenue Escadrille Normandie-Niemen (Case 151) 13397 Marseille Cedex 13, France. Department of Histology, Federal University of Sao Paulo, Sao Paulo CEP-0402,3, Brazil
Summary
The ultrastructural distribution and quantification of calcium in mast cells prepared by anhydrous processing was investigated by energy-filtering transmission electron microscopy (EFTEM) using a Zeiss 902 electron microscope. Optimal conditions for calcium detection were determined using inorganic (calcium phosphate) and organic (calcium-loaded chelex beads) standards with known amounts of calcium. Electron energy-loss spectroscopy (EELS) revealed calcium at the L-2,3 edge and also at the M-2,3 edge for all specimens examined. Comparison with X-ray microanalysis confirmed the results obtained with EELS. Electron spectroscopic imaging (ESI) was applied for mapping calcium both in standards and in cells and we showed that mast cell granules were the main site of calcium localization. Although, results have shown that a combination of analytical techniques is required to obtain reliable results.
Journal of Microscopy, Vol. 173, Pt. 3, March 1994, pp. 219--225.
CONTRAST IN THE TRANSMISSION MODE OF A LOW-VOLTAGE SCANNING ELECTRON MICROSCOPE
by U. GOLLA, B. SCHINDLER & L. REIMER, Physikalisches Institut, Universitat uMnster, Wilhelm-Klemm Str. 10, 4400u Mnster, Germany
Summary
The contrast thicknesses (x_k) of thin carbon and platinum films have been measured in the transmission mode of a low-voltage scanning electron microscope for apertures of 40 and 100mrad and electron energies (E) between 1 and 30keV. The measured values overlap with those previously measured for E (more than or equal to 17keV) in a transmission electron micro-scope. Differences in the decrease of x_k with decreasing E between carbon and platinum agree with Wentzel--Kramer--Brillouin calculations of the elastic cross-sections. Knowing the value of x_k allows the exponential decrease in transmission with increasing mass-thickness of the specimen and the increasing gain of contrast for stained biological sections with decreasing electron energy to be calculated for brightfield and darkfield modes.
Journal of Microscopy, Vol. 173, Pt. 3, March 1994, pp. 227--237.
MINIMIZING SAMPLE EVAPORATION IN THE ENVIRONMENTAL SCANNING ELECTRON MICROSCOPE
by R. E. CAMERON & A. M. DONALD, Polymers and Colloids Group, Cavendish Laboratory, Madingley Road, Cambridge CB3 0HE, U.K.
Summary
The ElectroScan environmental scanning electron microscope (ESEM) enables wet samples to be observed by eliminating air but allowing water vapour into the sample chamber. However, evaporation from, and condensation on, the sample may occur during the pumpdown sequence used to reach this state, which means that the sample may not be in its natural state when viewed if due care is not taken. In this paper, the pumping system of the ESEM is described mathematically and expressions are derived for the evaporation and condensation. This treatment is then used to calculate the optimum pumpdown sequence. The importance of using the optimized procedure is illustrated by micrographs of fat emulsions.
Journal of Microscopy, Vol. 173, Pt. 3, March 1994, pp. 239--243.
MORPHOMETRIC ASSESSMENT OF SURFACE CONFORMATION AS AN ESTIMATE OF ROUGHNESS
by D. A. SILAGE & G. R. BARAN, Departments of Electrical Engineering and Operative Dentistry, Temple University, Philadelphia, PA 19122 U.S.A.
Summary
One of the standard methods for assessing the roughness of a material subjected to wear uses the surface arithmetic means and root-mean-square deviation. However, these parameters often do not provide a qualitative assessment of the difference in materials worn under the same conditions of load and elapsed time. The profile and surface roughness parameters are frequently inconsistent. Such measurements are also required to determine the wear characteristics of various materials under different conditions. A morphometric assessment of wear characteristics, based on the surface area fraction of localized deviations in the surface texture and stress fractures, is provided, and clearly indicates the observed difference.
Journal of Microscopy, Vol. 173, Pt. 3, March 1994, pp. 245--256.
VOLUME AND SURFACE AREA MEASUREMENT OF VIABLE CHONDROCYTES IN SITU USING GEOMETRIC MODELLING OF SERIAL CONFOCAL SECTIONS
by F. GUILAK, Musculo-Skeletal Research Laboratory, Department of Orthopaedics, Health Sciences Center T18-030, State University of New York at Stony Brook, Stony Brook, NY 11794-8181, U.S.A.
Summary
This study describes a technique for noninvasive determination of the surface area and volume of chondrocytes using the confocal scanning laser microscope, and the fundamental limitations associated with its application. Using geometric modelling principles, an isointensity surface contour was formed from a series of optical sections recorded with the confocal microscope. Using a combined surface- and volume-based algorithm, the surface area, volume and other morphometric descriptions were calculated from a polygonal description of the cell surface. The high image contrast required for repeatable identification of the cell border was achieved through the use of a fluorescent dye, which was excluded from cells by an intact membrane. Calibration results indicated that the theoretical modelling algorithm is relatively precise when applied to simulated convex (ellipsoidal) cells, with overall errors of less than 0.5% in surface area and volume measurements. When applied to low-noise, high-contrast volume data recorded on the confocal microscope, typical coefficients of variation of 2--4% were determined for length measurements, 2--5% for volume measurements and 3--6% for surface area measurements either for latex microspheres or for chondrocytes. While the precision of the method is comparable to standard histological techniques, its accuracy is difficult to assess, as systematic errors are unpredictable and may be introduced from several sources.
FOR DETAILED INSTRUCTIONS TO AUTHORS, PLEASE CONTACT DR GILLIAN WILSON, THE JOURNAL OF MICROSCOPY, 37/38 ST CLEMENTS, OXFORD OX4 1AJ, UNITED KINGDOM. TELEPHONE +44 865 248768, FAX +44 865 791237, EMAIL RMS-at-UK.AC.OX.VAX.
NB. PAPERS FROM NORTH AMERICA AND CANADA MAY ALSO BE SUBMITTED TO PROFESSOR DAVID WILLIAMS, MATERIALS SCIENCE & ENGINEERING, LEHIGH UNIVERSITY, BETHLEHEM PA 18015-3195, USA. FAX +1 215 758 4244, EMAIL DBW1-at-LEHIGH.EDU.
Yes, I've thought since being made aware of MOSAIC that a server with images would be a wellcome addition to the net. I'd be willing to get involved. J.T. Stanley Oregon Graduate Institute Portland OR
On Wed, 16 Feb 1994 WALCKSD-at-ml.wpafb.af.mil wrote:
} This is an addenum to my previous suggestion. } } One of our computer gurus here at the lab suggested that we already have } the capability of sending digital images over the microscopy server, } just uuencode the graphic image and put a header in it saying what it } is. } } Nestor, will this work? Can we standardize such a transmission of an } image? Can it be tacked on to the end of the Email text? } } } -Scott Walck } } Dear Dr. Walck: The question you should also ask is what resolution will be available if you send images. In North Carolina we will soon be connected to a network that will allow high resolution images to be relayed around the state.
If you get further information about this and the cost of doing it , I would be very interested. Nina Allen, wake forest university
Reply_ RE} } Digital micrographs on I Reply from: John Mansfield North Campus Electron Microbeam Analysis Laboratory University of Michigan 2455 Hayward Ann Arbor MI 48109-2143 Phone: (313)936-3352 FAX: (313)936-3352 jfmjfm-at-umich.edu or John_Mansfield-at-mse.engin.umich.edu The anonymous ftp server at the University of Michigan could be a good site for this. I have a directory on feebie.engin.umich.edu for storing MSA & MAS related stuff, it is /pub/MSA+MAS. You can upload stuff to /pub/incoming and then email to me so I can move it to the MSA+MAS directory. If it is left in incoming it is liable to be deleted if the system is rebooted. (incoming is essentially a /tmp directory). If we got a large number of images I would archive them to optical disk or tape and then make them available on an MSA+MAS CD ROM. I was thinking of also making available on this disc archived messages sent to this list and the sci..techniques.microscopy newsgroup. I have been working with some peopl here to make the freebie ftp server accessible from Gopher and Mosaic. It seems that there may be some security questions, however. I am still working on it. Takes time and I dont seem to have enough hours in the day lately!! Cheers Jfm.
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This has been planned for the microscopy server, however, we have been waiting for our "new" computer to arrive, be installed & tested. The old system which we are currently running on is not worth spending any more $$ on and will be sacrificed to the silicon god in the near future. I expect the new system to be up, running and debugged within 2 months time.. Hopefully the transition will be seemless, but expect a few hickups. Just bear with the current setup a little bit longer.
Scott W asked about Emailing images over the server using uuencode.
The answer is yes it will work, however......
DONOT UNDER ANY CIRCUMSTANCES DO IT!!!!
It will jam up/fill all subscribers Email tremendously. Their mail boxes will fill and they (and I) will scream bloody murder. If someone wants a copy of an image they should contact you directly for it. If you want to make a particuliar image available to the subsribers for comment then wait until the new system is up an running and I will see how to most apprropriately (and simply) make this option avaiable to those who want to see the data. There are literally hundreds of subscribers who would not be interested in seeing your data, while maybe only a few dozen would be interested.
Also uuencode tends not to be universally accepted. Lots of users of this server donot have the capability to decode uuencoded data. That is a UNIX based system encoding many PC and Mac and Vax's which access this system will not have the appropriate translators (I note that they do exist however). Thus if you arbitrarily chose to send the data via a format that many users can't even read then it would be a waste of everyones time.
BTW this system is a VAX and not a UNIX machine and although we can translate uuencoded data we prefer alternatives which preserve more information, particuliarly if the data comes originally from a Mac or PC.
Message-Id: {MAILQUEUE-101.940217072125.480-at-parmly1.parmly.luc.edu} To: microscopy-at-anlemc.msd.anl.gov
} From: "Nestor J. Zaluzec (708)-252-5075, -4964" } Scott W asked about Emailing images over the server using } uuencode. } } The answer is yes it will work, however...... } } DONOT UNDER ANY CIRCUMSTANCES DO IT!!!! } ... An ignorant question, for when the new server is up: Is it possible to have mail sublists, say neuron.microscopy-at-..., that everyone on micrscopy-at-... could have access to, but not necessarily automatically get, as we do from this list? Thumbnails of images could be posted to the sublists for anyone to browse, and then contact the source of the thumbnail for the full image. This might make images available to the largest number of people, without overloading the server (?) or people's mailboxes. Phil Oshel
The University of New Mexico has an opening for a full-time position of Research Technologist V in the Transmission Electron Microscopy Laboratory in the Department of Earth and Planetary Sciences. The laboratory have a JEM 2000FX scanning transmission electron microscope with Noran EDS attachment and a newly installed JEM 2010 high resolution microscope with a Link ultra-thin window EDS and advanced image processing systems (including both Gatan live-rate TV system and slow-scan camera as well as IBM PC compatible and Macintosh computers). The laboratory also has a darkroom and full TEM sample preparation equipment. The responsibility of the technologist (under the supervision of the laboratory manager) includes but not limited to: routine maintenance of the laboratory and all the equipment (electron microscopes, ion mill, carbon coater, dimpler, diamond saw, microtome and darkroom equipment, etc.); supervise service engineers; train students, research staff and faculty on the use of microscopes and laboratory equipment; purchase consumable parts and supplies for the laboratory; participate in the research projects by preparing TEM samples, performing TEM/AEM analysis, preparing micrographs, slides and technical reports. The qualified candidate should have at least a BachelorÕs degree in science and engineering (Master's degree desirable) and four years of experience in a TEM laboratory (two years with MasterÕs degree). Experience on TEM maintenance and research projects using analytical and high resolution electron microscopy, knowledge with both IBM PC and Macintosh computers are desirable. The salary range is from $22,400 to $30,800/year depending on the qualifications. Application with a cover letter and resume must be received by Dr. Lumin Wang, Department of Earth and Planetary Sciences, University of New Mexico, Albuquerque, NM 87131-1116, by February 25, 1994.
The answer is maybe. We will also be getting the newgroup running and hopefully digest mode (?). It's just too early to say yes or no. The Newgroup function will effectively act as a sublist.. Please hold off on questions about the newsgroups etc.. for awhile, I've just gotten back from a meeting and have a bunch of catchup work to do, I will post details and information as new options are added to the server system. Again it will be ~ 2months before things are likely running...
A new professor in our department wants to purchase a nice used dissecting/operating microscope . I need names of places where one can purchase used microscopes.
Thanks for any help, nina allen, biology, wake forest university
Nina Allen asks were to buy a nice used micrscope. I am afraid I can't help with that. However, I am involved in getting some wonderful scopes from China. We are getting all types, from low power inspection, to phase contrast and darkfield 2000X trinoculars.
In general, the prices are lower than used Nikon/Zeiss/Olympus, etc. The quality is great. Most of these types have never been exported.
I am not ready to supply anything, but will have a sheet of sorts in about a week. If you would like to know more, send me a note. I am excited about the qualtiy/price of these devices.
Excuse the misspelled "where" in the first line herein. Slipped an "h".
Here is a summary of the recent discussion concerning magnetic specimens in a TEM:
TEM work on steel specimens can be very difficult, because they are almost always magnetized. You may be able to reduce the magnetic inhomogenieties a bit by passing the specimens through a strong AC field, a process industrially known as 'degaussing'. I believe degaussing coils can be purchased from most electronics supply stores that deal in the television market. In use, you insert the specimen into the center of the coil and withdraw it very slowly, with the AC current flowing through the coil.
Besides doing what Robert Keller suggests, you should note the objective lens current for a non-magnetic sample and set the lens to the same value. Then you can bring the specimen to the eucentric height by noting when the image is at the minimum contrast condition.
Russell Cook Electron Microscopy Center for Materials Research Argonne Natonal Laboratory Argonne, IL
Dear Colleagues,
Does anybody know an accurate value for the lattice parameter of GaAs at 120K (or 100K) ?
What are people using as a standard for determining the point spread function when the image is visualized with bright field (transmitted) light at high (1.3-1.4) numerical aperture? All of the articles I've seen involve fluorescence microscopy. Thanks for any leads.
Message-Id: {9402221505.AA29320-at-mogate.sps.mot.com} To: WALCKSD:ml:wpafb, SCHELTFJ:ml:wpafb Subj: Imaging Magnetic Materials Orig-Author: {finger Displays information about a user {bandaru-at-gandalf.rutgers.edu} }:ddn:wpafb ----------------------------------------------------------- A request for suggestions from those having experience with the Imaging of Magnetic Materials( Fe based). Any ideas on how to correct for the OL astigmatism , or good references to the above will be greatly appreciated. Please mail your requests to bandaru-at-gandalf.rutgers.edu Thanks in advance, Prabhakar R. Bandaru
The address for the company that manufactures the automatic dodging enlarger is: Logetronics Corporation, 7001 Loisdale Road, Springfield, VA 22150 (703) 971-1400
The company that repaired our enlarger recently is: Egoltronics Corporation, 7036 Tech Circle, Manassas, VA 22110 (703) 751-5522
LogEtronics has been bought out by another company (I believe) and has changed the name to Egoltronics Corp. Address is: 7036 Tech Circle Manassas, Virginia 22110 703-335-1501 (fax) 703-335-1234
I need to attach a 3-8 nm colloidal gold [preferably 5nm] particle directly to the primary antibody. Does anyone know of a company or companies that will do this with client's antibody?
Message-Id: {9402222328.AA20467-at-riker.ml.wpafb.af.mil} To: WALCKSD:ml:wpafb, SCHELTFJ:ml:wpafb Subj: Imaging Magnetic Materials Orig-Author: {finger Displays information about a user {bandaru-at-gandalf.rutgers.edu} }:ddn:wpafb ----------------------------------------------------------- A request for suggestions from those having experience with the Imaging of Magnetic Materials( Fe based). Any ideas on how to correct for the OL astigmatism , or good references to the above will be greatly appreciated. Please mail your requests to bandaru-at-gandalf.rutgers.edu Thanks in advance, Prabhakar R. Bandaru
You might try E-Y Laboratories. They specialize in colloidal gold conjugates (tracers, antibodies, etc), and I think (!) I remember that they would conjugate your reagent for you. Anyway, it might be worth checking:
E-Y Laboratories, Inc. 107 N. Amphlett Blvd. San Mateo, CA 94401 415-342-3296
Good luck
David Morilak morilak-at-cmgm.stanford.edu -------
Message-Id: {9402231439.AA22563-at-riker.ml.wpafb.af.mil} To: WALCKSD:ml:wpafb, SCHELTFJ:ml:wpafb Subj: Lacquer as protective coating Orig-Author: {"Joel Hoehn" {Hoehn-at-maroon.tc.umn.edu} }:ddn:wpafb ----------------------------------------------------------- I am trying to protect the front side of some Fe-3%Si while polishing from the back side in a tenupol using 95% acetic / 5% perchloric solution. I have heard of a sort of lacquer used for the procedure and would like to know if anyone could tell me of a supplier of such a compound. From what I understand, the usual microscopy type suppliers (i.e. ted pella, spi, etc.) do not carry this sort of product, but some industrial application type companies do.
Any leads would be helpful.
Joel Hoehn e-mail: Hoehn-at-maroon.tc.umn.edu Dept. of Chem. Engg & Materials Science phone#: (612) 625-1018 University of Minnesota 421 Washington Ave. S.E.
Message-Id: {9402231451.AA22632-at-riker.ml.wpafb.af.mil} To: WALCKSD:ml:wpafb, SCHELTFJ:ml:wpafb Subj: Magnetic specimens in TEM Orig-Author: {jrmicha-at-saix367.sandia.gov (MICHAEL, JOSEPH)}:ddn:wpafb ----------------------------------------------------------- I spent 6 years at BEthlehem Steel's research department using TEM and STEM to study steel microstructures. The most important thing that can be done to minimize the difficulty of magnetic specimens is to make the specimen blank as thin as possible. Normally, we would electrochemically polish the specimen blanks to a thickness of about 50 microns before we would jet polish them to perforation. This minimizes the amount of magnetic material within the field of the lens.
We tried de,agnetizing the specimens, but found that the fields generated within the coil usually destroyed the thin area of the specimen.
} } We have a Kevex 8000 PDP 11 DEC EDS computer with horrible 8 inch } Bernoullis. They are expensive, unreliable. We have two questions. } } 1) Has anyone successfully used kermit to communicate with a pc from this } type of computer? } } 2) Is there a file translation utility that would transfer the intensity } vs. energy to an ascii } format? } We have a similiar system with some of the same needs. We replaced the 8 inch Bernoullis with twin Bernoullie 44MB drives and improved things considerably. If you do this you will have to replace a chip on one of the boards in the computer. This may be the only item you have to purchase from Kevex.
As for kermit, we have placed kermit on the system with success. We had a couple of problems doing it though. The first was getting kermit on a proper media. The second was adding it to the menu. We purchased a program called TIFMKR from Kevex and had them load Kermit on the same disk. TIFFIT/TIFMKR converts Images from Advanced Digital Imaging, Screen Images, and EDS images to TIFF format files that can be transfered from the DEC to your PC using Kermit. Unfortunately the conversion is fairly low resolution and not practical for the Advanced Imaging images. Kevex also wants a lot of money for this package but they will negotiate. Check with Kevex for specific information.
There is a red lacquer MICROSTOP used for 'the window pane' method and to protect tweezers, etc. while chemically thinning samples. The lacquer was thinned with acetone. We had it at North Carolina State. It had been in the lab of Professor Ray Benson, at (919) 515-2706.
You could also try photoresist. It should be very easy to obtain at U of Minn, is chemically resistant, and removable with acetone or oxygen plasma. Some types of photoresist clean up from samples easier than others.
Just a bit of information for those of you that are interested in details. The Microscopy Listserver just recorded its 750th subscription request. Seems like it was a reasonable idea after all. ;-)
If you add up the total number of Email transmissions that the system has handled over the last 5 months we have sent out over 250,000 individual messages. No wonder my computer is getting tired!
Honors & a beer (or whatever he would like when I see him) for being the 750th subscriber go to:
Mike Bench Dept. of Chem Eng & Mat. Sci Univ. of Minnesota
============================= Nestor J. Zaluzec ANL EM Center
I am passing on the following message for discussion:
Electron Microscope Center Mississippi State UniversitySubject: Formvar films cast on glass
Question: Our laboratory staff has tried various methods over the years to get the formvar film to release from glass slides on a regular, consistent basis while producing formvar-coated TEM grids.(Problem: the films sometimes do not release at all, sometimes release too wrinkled to use, etc.) We use formvar prepared in chloroform, and make use of an Effa Film Caster Apparatus (Ernest F. Fullam, Inc.). I would appreciate any suggestions which might help alleviate this dilemma.
Can anyone tell me the mechanism of UAc fixation and staining of wood components, specifically lignin, for EM? I am interested in learning about the chemical groups of the sample that may be involved. I am also interested in learning about the structure and chemical groups of lignin and have not been able to find any recent information.
Like many people we have been using double distilled water for washing etc in immuno gold protocols. We will soon have much easier access to an ultrapure (type 1, 18 M ohm) water system. Does anyone know if this will cause any difficulties or is it essentially the same.
Also some people manage to get away with "straight" distilled water, any comments?
Ian Hallett EM Facility HortResearch Auckland New Zealand
Regarding the request about water purity and immuno staining.
I think it very much depends on your local water source. We have used normal distilled water without any problems at all and have in fact obtained very good localizations. Some of the researchers here use the ultra pure water but that is mainly for in situ hybridization, although light microscopy immuno work has been carried out with the ultra pure water without any hassles as well.
Peter RichardsPeter D. G. Richards Dept Anatomy and Cell Biology UCT Medical School Observatory 7925 RSA Tel: 021-406 6285.
Message-ID: {MAILQUEUE-101.940224084456.800-at-anat.uct.ac.za} To: microscopy-at-anlemc.msd.anl.gov
Regarding the recent query about formvar coating passed on from Bill Munroe.
I do all my coating in a similar way but without the film caster. I find that the glass slides I use for the film have to be clean but not ultra clean. If the glass is to clean then there are problems with removal of the film on to the water surface, as the formvar sticks well to the glass, as Bill has obviously found.
When the glass slide is to dirty then obviously a poor film results, however with a slight "greasyiness" the films come off well and evenly with no wrinkling.
Peter RichardsPeter D. G. Richards Dept Anatomy and Cell Biology UCT Medical School Observatory 7925 RSA Tel: 021-406 6285.
Bill, I coat glass slides with Formvar on a regular basis and have had similar trouble in the past. I have solved it completely, however, by a very simple means. Just wipe a finger across your forehead and lightly coat the side of the slide on which the released coating will be (works best towards the end of the day, obviously). Then with a teatowel or the corner of your lab coat, buff the slide moderately clean. ie. leave a microscopic layer of body grease on the surface. This ensures that the Formvar will release if you have scored all around the slide edges with a sharp razor blade. No need for fancy equipment - just dip the slide slowly, at an angle so one corner releases first, into a wide mouthed vessel of distilled water. The cast film with its grids is then very easily picked up with Parafilm rolled across the film and lifted.
Good luck
Nikki Watson
Dr N.A. Watson Department of Zoology University of New England Armidale, NSW, 2351 AUSTRALIA Fax: 067 711 869 Phone: 067 732181
Do Kontron still exist? Are they now part of a bigger company? Do they still make inage analysis systems? Does somebody have a US phone number? Alwyn Eades Center for Microanalysis of Materials University of Illino
Our trick for getting consistent release ofFormvar films is to spray the slide with Fantastic or 409 spray cleaner and polish the slide dry without rinsing. The residual detergent film allows easy release of the film and it is generally of good quality. In our humid climate we also store the solution and the posdre in a sealed container with silica gel. ***************************** * Greg Erdos * * Director, ICBR EMCL * * University of Florida * * Gainesville, FL 32611 * * gwerdos-at-gnv.ifas.ufl.edu * * 904-392-1295 * *****************************
Do Kontron still exist? Are they now part of a bigger company? Do they still make inage analysis systems? Does somebody have a US phone number? Alwyn Eades Center for Microanalysis of Materials University of Illino
Message-ID: {MAILQUEUE-101.940224165025.288-at-anat.uct.ac.za} To: microscopy-at-anlemc.msd.anl.gov
Further to the requests about water and immuno staining.
Some of the local proponents of immuno work out here are using only local tap water without any purification. Certainly the results are good and there does not appear to be any background problems as a result. Peter D. G. Richards Dept Anatomy and Cell Biology UCT Medical School Observatory 7925 RSA Tel: 021-406 6285.
When all else fails: we use nose grease rather then forehead grease (of course, if you are a woman or a man wearing makeup this doesn't work at all ). If nose grease doesn't work, we score the slide normally and place it face side down over a 65mm plastic culture dish containing a few drops of hydrofluoric acid (be careful! this is bad stuff) for a few seconds. Then we exhale on the slide as usual and generally the films strip easily. Too long of an exposure to HF acid results in poor films so you have to practice with it depend upon humidity, barometric pressure, how the stars are aligned, and so on.... If all this doesn't work, we try again tomorrow ! Good luck.
We use ethylene dichloride as the solvent. The slides are cleaned first in ethanol until the grease-streaks disappear. After drying the slide for a minute or two, we score the bottom and side edges (45 degree angle to face of slide) with a razor blade, then we also score the face of the slide on all four sides with a razor blade. I score several times along the bottom face. If one set of score marks fails, the other is likely to succeed. We usually use Gold Seal Microslides but have have used other brands as well. Problems are rare.
Rod
On Wed, 23 Feb 1994, Richard F. Kuklinski wrote:
} I am passing on the following message for discussion: } } From: Bill Monroe } Electron Microscope Center } Mississippi State UniversitySubject: Formvar films cast on glass } } Question: Our laboratory staff has tried various methods } over the years to get the formvar film to release from } glass slides on a regular, consistent basis while producing } formvar-coated TEM grids.(Problem: the films sometimes do } not release at all, sometimes release too wrinkled to use, } etc.) We use formvar prepared in chloroform, and make use } of an Effa Film Caster Apparatus (Ernest F. Fullam, Inc.). } I would appreciate any suggestions which might help } alleviate this dilemma.
The ultrapure water water systems clean out particles, organics and ions--- they do not, however, degas the water. Small bubbles can be a problem in photography (pin holes in emulsion) or in putting grids underwater on filter paper(small bubbles form on the grids and make them hard to manipulate) in preparation to picking up carbon films. We do not consider any of these problems serious and use ultrapure water all the time!
In Center for HREM, Arizona State University (ASU), we read all EDS and EELS data in a Vax system by "RT 11". Then we can convert these binary data using a program "convascii" that was written by our computer manager Mr. Paul R. Perkes, into a ascii files. Then read these files either from Mac or PC.
If you are interested in this "convascii" program, you have to contact Mr. Paul R. Perkes email:smtp%"perkes-at-asuhrm.la.asu.edu"
Lacquer as protective coating In our lab we use a product called Miccroshield 1 ( Miccroshield 2 is its solvent ) as a protective coating during etching of silicon crystals in 5% HF + 95% HNO3. It's a lacquer that is brush on, drys quickly and is easily removed. From the MSDS here's the manufacturer info:
TOLBER DIVISION/PYRAMID PLASTICS INC. 220 WEST 5TH STREET HOPE, ARKANSAS 71801 # (501) 777-5759
The Microshield Lacquer has been used for many years with our Model 550 Electropolisher and Model 550 Chemical Jet Polisher. It is acetone soluble and is available in an 8 ounce kit. Please reference P/N 02-01890-01.
We use formvar dissolved in Dichloroethane (0.6%). We cast onto slides which have been polished in a drop of liquid detergent, leaving a very thin film over the slide and dried 60 C for 10 minutes or so. After dipping the slide in the formvar solution we leave to dry for 10 minutes or so befor stripping, this gives much more consistant release.
Ian Hallett EM Facility HortResearch Auckland New Zealand EM Facility
Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii" To: Microscopy-at-anlemc.msd.anl.gov
} Question: Our laboratory staff has tried various methods } over the years to get the formvar film to release from } glass slides on a regular, consistent basis while producing } formvar-coated TEM grids.......
From David Bentley:
When we have trouble with the film not releasing, we briefly 'polish' the slides with a Kimwipe (which may actually end up depositing a thin film of grease from our fingers on the slide); if this does not work, we use Kimwipes to clean the slides with Windex, which seems to leave a 'parting layer' of some kind on the glass. We use Gold Seal slides from Clay Adams (cat. # 3052). The films generally part well when the slides are _fresh_ (e.g. less than three months after the seal of the _inner_ mylar bag of the shipping container is broken). The slides than begin to exhibit patchy 'frosting' or 'fogging' which is visible when a slide is held up to the light; the film seems to stick to the fogged areas. This fog eventually covers the entire surface of the slide, and can be removed by acid (5% HCl/95% ethanol) cleaning followed by Windex.
In the last mail alwyn eades said: } } } The general view is that the best lacquer is "Lacomit", a bright red } lacquer. } It is resistant to almost all polishing solutions but is removed easily by } normal solvents, for example acetone. The red colour helps to see whether } it has been cleanly removed. } Unfortunately, it is not avaiable in the USA. As far as I know it is still } avaiable in England. If you get someone to bring you a large bottle, it } will last until there are no more electron microscopes.
If you do get someone to do this, either get them to bring a bottle of 'lacomit thinner' as well - over time the undiluted stuff tends to turn into a jelly which needs occasional thinning. Also, when you remove it, (I usually use acetone), make sure that the wash is completely clear with no trace of pink; if this is not done, a residue is left which is almost impossible to remove and sits all over your thin area. 8).
Richard Beanland, Dept. of Mat. Sci. and Eng., University of Liverpool, P.O. Box 147, Liverpool, England.
I'm not sure what my options are regarding the microscopy list. I previously tried to unsubscribe by sending a message (UNSUBSCRIBE MICROSCOPY wacb-at-aplcomm.jhuapl.edu) to listserver-at-anlemc.msd.anl.gov, but still I get swarms of email from the list. Perhaps this (MICROSCOPY) was the wrong name to use for the list. Now, in a more circumspect and appreciative mood, I would like to see what other options I have for achieving a more managable flow to me from the list. Some lists have a "digest" mode that collects all of the messages to the list on a single day into a single mailing to the list members, and this would probably be appropriate for me here.
And so, I ask, how and where do I send a message that will elicit a list all of the commands and options that exist for list members? What is the command syntax that will do this for me? "help" or "index" etc. are frequently used in some lists to get the kind of response I have in mind.
Sorry to send this directly to you, but I don't know how else to address this sort of bootstrapping (or maybe I should say bootstripping) problem.
Message-Id: {MAILQUEUE-101.940224155128.448-at-ahabs.wisc.edu} To: microscopy-at-anlemc.msd.anl.gov
While greasing the slide this way works great, anyone who subsequently grows cells on the grid may need to be concerned about the grease transferring to the formvar. I have found that just polishing the slide with a kimwipe (not too much or the formvar will stick) is usually enough to get it to release.
Bill, I coat glass slides with Formvar on a regular basis and have had similar trouble in the past. I have solved it completely, however, by a very simple means. Just wipe a finger across your forehead and lightly coat the side of the slide on which the released coating will be (works best towards the end of the day, obviously). Then with a teatowel or the corner of your lab coat, buff the slide moderately clean. ie. leave a microscopic layer of body grease on the surface. This ensures that the Formvar will release if you have scored all around the slide edges with a sharp razor blade. No need for fancy equipment - just dip the slide slowly, at an angle so one corner releases first, into a wide mouthed vessel of distilled water. The cast film with its grids is then very easily picked up with Parafilm rolled across the film and lifted.
Good luck
Nikki Watson
Dr N.A. Watson Department of Zoology University of New England Armidale, NSW, 2351 AUSTRALIA Fax: 067 711 869 Phone: 067 732181
} I am passing on the following message for discussion: } } From: Bill Monroe } Electron Microscope Center } Mississippi State UniversitySubject: Formvar films cast on glass } } Question: Our laboratory staff has tried various methods } over the years to get the formvar film to release from } glass slides on a regular, consistent basis while producing } formvar-coated TEM grids.(Problem: the films sometimes do } not release at all, sometimes release too wrinkled to use, } etc.) We use formvar prepared in chloroform, and make use } of an Effa Film Caster Apparatus (Ernest F. Fullam, Inc.). } I would appreciate any suggestions which might help } alleviate this dilemma. }
This always works for me:
I use formvar in ehthylene dichloride and if it is over a month old I discard it. I have real problems with Gold Seal micro slides but no problems with Fisher pre-cleaned micro slides, cat no. 12-552. I wipe the slide with a kimwipe but I don't over do it. I don't have a fancy film caster, I just dunk the slide into .25 to .4% w/v soln. for a few seconds and then let it air dry for a minute. I then run the side of my forceps lightly around the edge of the slide, not on top of the slide but on the angle made by the top and side of the slide. Next, I breath on the slide to form a moist layer (I have always done this, don't know why). Then I gently dip slide straight down while holding it at an angle of about 45 degrees. Works every time, never wrinkles. Gold Seal slides would not release formvar. Some people like to use glass strips from the microtome but I have not tried this.
Rick A. Harris Supervisor, Microsopy Facility Evolution and Ecology, Storer Hall Univer. of Calif., Davis
I have read this discussion with interest. There seem to be extremely many methods with all the same principle.
The main idea seems to be that you have to make the surface of the object glas hydrophilic with some method. The usual way is to coat the glas with a "monolayer" of some polaric molecules, e.g. detergent, as many of you have suggested. This layer then attracts the water molecules and they are slipping in between the glas and the formwar thus removing the formwar- layer. There are many different hydrophilic commercially awailable solutions with which you can even adjust the strength of hydrophilicity - as well as the opposite = hydrophobicity.
This monomolecular layer is also good against microscopic pits in the glas surface because it covers the pit to some extent.
- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - Jouko K. Maki Navigare necesse est... Laboratory Manager, Ph.D. Laboratory of Electron Microscopy Kiinamyllynkatu 10 FIN-20520 TURKU FINLAND University of Turku Tel.: + 358 21 633 7318 INTERNET: jouko.maki-at-utu.fi FAX.: + 358 21 633 7380
Plant cell walls become highly fluorescent after fixation with glutaraldehyde. A student here used that method to estimate total cell wall by morphometrics. I think he left them in 10% glut overnight. ***************************** * Greg Erdos * * Director, ICBR EMCL * * University of Florida * * Gainesville, FL 32611 * * gwerdos-at-gnv.ifas.ufl.edu * * 904-392-1295 * *****************************
In reply to your deconvolution questions: 1) The maximum entropy sol- ution is not always the smoothest; in particular, the m.e. solution for dif- fraction gives peaks which get sharper as the phase set is extended. Since, for your example, the maximum resolution is limited by the probe diameter, this effect may not occur; however, if the solution tree gives maximum likelihood for a set of structures which are not smooth, there is nothing which would guide the solution to a smoother structure. That is, suppose there are two structures the first of which is smooth and nearly correct and the second of which is rough at a scale smaller than the probe diameter and whose envelope is even more nearly correct. M.e. would follow the path of the rougher structure, and would not necessarily smooth it out. 2) Leaving aside the question of whether atoms are delta functions--especially when they are at the sum of the Van der Walls radii apart--maximum entropy can find such a structure and will find a structure whose envelope follows the contact points of the atoms with the probe. The often-published STM images of a silicon surface demonstrate what one might expect. It is not a bad idea, however, to include atomicity or other constraints explicitly, and, I believe, this can be done easily in a m.e. calculation. I hope this helps.
IgGs & Gold Dear Dr Ruben, I tried to send this message out on the bulletin board but I seem to be having problems with the connection. I hope that this will answer some of your questions. Feel free to call me if you want more information etc.
There are many reviews out in the literature covering the different methods for producing colloidal gold probes and then coupling them to proteins, including IgGs (look under Jan De Mey, Jan Leunissen, Ralph Albrechts). The methods are easy to follow and easy to do. If you would prefer someone else to do this then I think the lab in Utrecht, Holland provides this service - the contact is George Postuma, Dept of Cell Biol. University of Utrecht, The Netherlands (phone number 011 31 30 507 654) - this is the lab where Jan Slot works, by the way. Coupling immunoglobulins to gold requires lots of protein so make sure you have large supplies of your antibodies, especially if you want to do double label experiments and need IgGs coupled to more than one gold size. If you only have limited supplies of antibody and are planning simple immunocytochemical experiments then it will be simpler and less expensive to use protein A-gold. This will bind to rabbit antibodies and a few other IgGs. However, mouse monoclonals, in general have poor or no affinity for protein A. In this case, buy an unconjugated rabbit anti-mouse and use this as a bridge (ie apply first antibody; rabbit anti-mouse or anti-rat etc; apply protein A gold). Contact me if you want to use the antibodies for double labeling. These are simple methods using bridging antibodies and protein A-gold. Paul Webster Yale School of Medicine (203) 785 5072 Good Luck.
In reply to your deconvolution questions: 1) The maximum entropy sol- ution is not always the smoothest; in particular, the m.e. solution for dif- fraction gives peaks which get sharper as the phase set is extended. Since, for your example, the maximum resolution is limited by the probe diameter, this effect may not occur; however, if the solution tree gives maximum likelihood for a set of structures which are not smooth, there is nothing which would guide the solution to a smoother structure. That is, suppose there are two structures the first of which is smooth and nearly correct and the second of which is rough at a scale smaller than the probe diameter and whose envelope is even more nearly correct. M.e. would follow the path of the rougher structure, and would not necessarily smooth it out. 2) Leaving aside the question of whether atoms are delta functions--especially when they are at the sum of the Van der Walls radii apart--maximum entropy can find such a structure and will find a structure whose envelope follows the contact points of the atoms with the probe. The often-published STM images of a silicon surface demonstrate what one might expect. It is not a bad idea, however, to include atomicity or other constraints explicitly, and, I believe, this can be done easily in a m.e. calculation. I hope this helps.
At the ANLEMC we also use Process Software's FTP & TCP/IP software on the Dec PDP 11's. We use this on both normal PDP 11's and also EDAX PV9900 systems (PDP 11/73 based). All systems transfer both images and spectra back and forth between Dec, Mac, and PC systems. To transfer images we send data in raw (binary) format and then use a variety of programs to import and read the data. For example we can read EDAX Image files directly into the NIH Image program for fast viewing and sorting. We can also transfer spectra both in binary (Edax) format or alternatively translate the spectra on the EDAX system into the MSA (Microscopy Society of America) format, which is a simple ASCII file. Once the data is in the MSA format it can be directly transported into any data analysis/spreadsheet progran on your favorite platform. The MSA format is in the public domain and can be downloaded from the EMMPDL.
I've been off line for the last 2 days, however, I did recieve your collective messages that some of you were having problems. the system should be fixed by now, but please keep me informed of problems whenever they occur. The lastest problem was relatively simple the disk was full and hence Email was either being rejected or sent out with some or all of the text of messages missing. Sorry about that.... :-(
As for the other questions about creating a sublists, digest modes, etc..... These have always been possiblities and I'm always open to the suggestions for improvements (no offense will be taken on anyones comments/suggestions for improvements). Basically these items are on my list of things to do, but as I mentioned earlier this month (or was it last month?) , I'm trying to procur a new system, install it debug and generally get it up and running and then decommission this computer (all in my spare time). Until I have these all completed, I'm going to have to preserve the status quo.
*************************************************************************** CALL FOR PAPERS ***************************************************************************
We are organizing a focused workshop entitled:
"INTERFACE FORMATION AND DYNAMICS IN LAYERED STRUTURES"
This workshop will be part of the Scanning Microscopy 1994 Meeting to be held in Toronto, Canada, 9-11 May, 1994.
The objective of this workshop is to bring together academic and industrial researchers working in the field of epitaxial heterostructures. Topics will include: -surface and interface energetics of defect formation -sensitivity of materials properties to interface imperfections -characterization techniques for interface studies -epitaxial growth modes -phase separation processes -point and line defect engineering -applications of mismatched materials.
The workshop has been organized on the same lines as the Gordon and European Science Foundation Research Conference format. Accordingly there will be much time for discussion. Moreover there will be a large number of invited speakers including:
G.C. Aers (NRC-Ottawa) J.-M. Baribeau (NRC-Ottawa) E. Bauer (U. Clausthal) D.K. Biegelsen (Xerox) D. Cherns (U. Bristol) A.G. Cullis (DRA-Malvern) C.B. Duke (Xerox) K. Eberl (Max Planck Inst.) L.C. Feldman (AT&T) E.A. Fitzgerald (AT&T) J.M. Gibson (U. Illinois) M. Grinfeld (Rutgers U.) D.E. Jesson (ORNL) B.A. Joyce (Imperial College) K.L. Kavanagh (UCSD) R.E. Mallard (BNR-Ottawa) B. Meyerson (IBM) B. Orr (U. Michigan) C.J. Palmstrom (Bellcore) H.E. Ruda (U. Toronto) M. Saran (Northern Telecom) L.J. Schowalter (Rensselaer) T. Tiedje (UBC) P.W. Voorhees (Northwestern U.) G.C. Weatherly (McMaster U.) Y.-N. Yang (U. Maryland) A. Zangwill (Georgia Tech)
The deadline for submission of contributed papers is 15 April, 1994.
For further information about the workshop please contact:
Doug D. Perovic Department of Metallurgy and Materials Science, University of Toronto, Toronto, M5S 1A4 Canada Tel: (416) 978-5635 Fax: (416) 978-4155 Email: perovic-at-ecf.utoronto.ca
To: WALCKSD:ml:wpafb, SCHELTFJ:ml:wpafb Subj: Problems & Question with the Email Orig-Author: {"Nestor J. Zaluzec (708)-252-5075, -4964" {ZALUZEC-at-anlemc.msd.anl.gov} }:ddn:wpafb ----------------------------------------------------------- Friday 2/25/94 ~ 11 pm
All Subscribers
I've been off line for the last 2 days, however, I did recieve your collective messages that some of you were having problems. the system should be fixed by now, but please keep me informed of problems whenever they occur. The lastest problem was relatively simple the disk was full and hence Email was either being rejected or sent out with some or all of the text of messages missing. Sorry about that.... :-(
As for the other questions about creating a sublists, digest modes, etc..... These have always been possiblities and I'm always open to the suggestions for improvements (no offense will be taken on anyones comments/suggestions for improvements). Basically these items are on my list of things to do, but as I mentioned earlier this month (or was it last month?) , I'm trying to procur a new system, install it debug and generally get it up and running and then decommission this computer (all in my spare time). Until I have these all completed, I'm going to have to preserve the status quo.
I am looking for microscopically orientated people who may be visiting South Africa in the next 6-12 months.
I have recently submitted and had accepted the DipRMS thesis, but am trying to assist the Royal Microscopical Society in finding a microscopist who would be able to convene a local viva board. The person would have to meet the Royal Microscopical Society's approval, as being a person able to perform such a function.
If you know of anyone or if you are able to assist yourself could you please contact me directly at
RETEP-at-ANAT.UCT.AC.ZA
or phone/write to me at the telephone number/address below.
PLEASE DO NOT respond via the Mailserver.
Peter D. G. Richards Dept Anatomy and Cell Biology UCT Medical School Observatory 7925 RSA Tel: 021-406 6285.
*************************************************************************** CALL FOR PAPERS ***************************************************************************
We are organizing a focused workshop entitled:
"INTERFACE FORMATION AND DYNAMICS IN LAYERED STRUTURES"
This workshop will be part of the Scanning Microscopy 1994 Meeting to be held in Toronto, Canada, 9-11 May, 1994.
The objective of this workshop is to bring together academic and industrial researchers working in the field of epitaxial heterostructures. Topics will include: -surface and interface energetics of defect formation -sensitivity of materials properties to interface imperfections -characterization techniques for interface studies -epitaxial growth modes -phase separation processes -point and line defect engineering -applications of mismatched materials.
The workshop has been organized on the same lines as the Gordon and European Science Foundation Research Conference format. Accordingly there will be much time for discussion. Moreover there will be a large number of invited speakers including:
G.C. Aers (NRC-Ottawa) J.-M. Baribeau (NRC-Ottawa) E. Bauer (U. Clausthal) D.K. Biegelsen (Xerox) D. Cherns (U. Bristol) A.G. Cullis (DRA-Malvern) C.B. Duke (Xerox) K. Eberl (Max Planck Inst.) L.C. Feldman (AT&T) E.A. Fitzgerald (AT&T) J.M. Gibson (U. Illinois) M. Grinfeld (Rutgers U.) D.E. Jesson (ORNL) B.A. Joyce (Imperial College) K.L. Kavanagh (UCSD) R.E. Mallard (BNR-Ottawa) B. Meyerson (IBM) B. Orr (U. Michigan) C.J. Palmstrom (Bellcore) H.E. Ruda (U. Toronto) M. Saran (Northern Telecom) L.J. Schowalter (Rensselaer) T. Tiedje (UBC) P.W. Voorhees (Northwestern U.) G.C. Weatherly (McMaster U.) Y.-N. Yang (U. Maryland) A. Zangwill (Georgia Tech)
The deadline for submission of contributed papers is 15 April, 1994.
For further information about the workshop please contact:
Doug D. Perovic Department of Metallurgy and Materials Science, University of Toronto, Toronto, M5S 1A4 Canada Tel: (416) 978-5635 Fax: (416) 978-4155 Email: perovic-at-ecf.utoronto.ca
I have always used gelatin capsules for embedding material in LR-White and I was wondering if anyone has tried using Beem capsules. If I dry the Beem capsules overnight in a 50 C vacuum oven, would this help? I'm currently doing immuno-labeling on Euplodes and would like to be able to spin the specimens down into a conical Beem capsule. Any suggestions? I appreciate any info on this method. Thanks, Phil Rutledge prutle1-at-gl.umbc.edu
This is a request for help. A zoology faculty member wants to count fiber types (muscle fiber cross sections, stained for I, IIA and IIB) and to obtain cross section areas for the fibers. If I understand the needs correctly, we will need software that will allow "sliding" two images and aligniment of the two for fiber type identification. Then, a more usual morphorometry of doing the X-sec. Defining the boundry by hand for the area determination is acceptable. Is there anyone who is doing this or has experience with muscle firber analysis that can recommend software for either the PC or Mac platform? I can capture the image into a PC system in my lab for them, save it in a standard format .tga or tiff etc., but have no microcomputer level analytical software. Any suggestions, vendors etc will be appreciated. Pardon the typo, live time VAX. Thanks David P. Campbell
We also ended up with a big mess when heat polymerizing LR White in polyethylene BEEM caps. However a polypropylene micro-fuge tube will work just fine. Only problem is that one must saw out the specimen or use doggie toenail clippers to cut off the end of the tube wherein lies the specimen and then tease it out. LR White does not bind to the polypropylene but it doesn't slip right out either. If you use the clippers be sure to do this inside your closed fist so that the specimen is not launched across the room never to be seen again. The clipped off tip must then be remounted on something appropriate for sectioning using super glue. Works best if you file done the cut surface so that it is smooth and makes good contact with the plastic stub such as those sold by Pella.
} Phil Rutledge asked about polymerizing LR-white in Beem capsules. We tried } drying at 60 C for 4 hours (haven't tried overnight). Results were } just as bad as with undried capsules. Poor polymerization of the resin. } We ended up with a real funky mess. } -Jay Jerome } jjerome-at-isnet.is.wfu.edu
I've never used LR-white, but I've talked with one of the distributors about its notorious polymerization properties. They told me that they think BEEM capsules are not suitable for polymerizing LR-white. The reason is that the capsules are too porous and permeable to water. Any water will interfere with polymerization. They have never had any trouble polymerizing, as long as they use gelatin capsules.
As I said, I have no experience (yet), but this might help others. I'd be interested in comments from those who use it successfully.
John chandler-at-lamar.ColoState.EDU Fort Collins, CO
Message-Id: {m0pbe6h-0000PNC-at-stjohns.ohsu.edu} Message-Version: 2 } To: microscopy-at-anlemc.msd.anl.gov
------------------------------------------------------------------------------ POSITION AVAILABLE ------------------------------------------------------------------------------
Research Associate postition open at the Department of Pathlogy in research related to cytoskeletal components of megakaryoscytes and the process of platelet fromation. Successful applicant will have a M.S. or equivalent, experience in electron microscopy and immunology techniques. Experience in standard biochemical and molecular biology techniques preferred.
Contact by phone or regular Mail:
Dr. Paula Stenberg, E.M. Director Department of Pathlogy, L113 Oregon Health Sciences University Portland, OR 97201
Message-Id: {m0pbeRw-0000P0C-at-stjohns.ohsu.edu} Message-Version: 2 } To: microscopy-at-anlemc.msd.anl.gov
Evan's Blue and Blood Brain Barrier
A general question for light level resolution. We use Evan's Blue circulated in vivo for 30 minutes to see if there has been a breakdown of the blood brain barrier. Animals are then perfused (4% para.) the brains sectioned with a vibratome (100 microns). Question is: If we dehydrate and mount in permount will the Evan's blue stay put or get leached out? Any experience with similar situation would be helpful.
Bob Kayton C.R.O.E.T. L606 Oregon Health Sciences Universtiy Portland, OR. 97007 503 494 2504 E-mail kayton-at-ohsu.edu
Message-ID: {MAILQUEUE-101.940302153004.576-at-anat.uct.ac.za} To: microscopy-at-anlemc.msd.anl.gov
Regarding the query about the azo vital dye Evans Blue.
I have not had any personal experiance with the dye but there should be no problem with dehydrating and mounting. Be sure to dehydrate in alcohols and xylene or equivelant.
Peter D. G. Richards Dept Anatomy and Cell Biology UCT Medical School Observatory 7925 RSA Tel: 021-406 6285.
Phil: our lab's experience with yeast, LR White and capsules: two ways: 1. small sample size: Place suspension of sample in LR White in a BEEM capsule, seal and centrifuge ( inside a tube, whose bottom is padded with cotton). Seal the BEEM INSIDE a large gelatin capsule. Available through drugstore or EM supply house. If you can't find a large enough gelatin capsule; use the OOO. Simply "trim" the beem capsule(that extra ridge where the cap goes on), so it will slide in.Then carefully seal the BEEM capsule inside two OOO gelatin capsule bottoms, that have been filled with LR White. A bit sloppy, but very effective. Works for us 100%. You just need to wear gloves and put down extra paper to catch any LR white drops, while you are filling and sealing 2. large sample size: leave material in OO capsules. Place these capsules inside a BEEM capsule, that has been cut to accomodate the capsules (The top half is cut away). Clinical centrifuge, as above.
hope this helps Louisa. Howard-at-dartmouth.edu
P.S. If you have access to a vacuum oven, you can use the BEEM capsules alone. This avoids the problems associated with porous BEEM capsules and the presence of oxygen.
Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii" To: MICROSCOPY-at-ANLEMC.MSD.ANL.GOV
} Is there anyone who is doing this or has experience with muscle firber analysis } that can recommend software for either the PC or Mac platform?
NIH-Image, a freeware image processing program for the Macintosh available from the National Institutes of Health at zippy.nimh.nih.gov, directory /pub/image, has facilities for image alignment (in several modes), area determination (including thresholding/density slicing to segment the regions of interest, and automatic counting, analysis and labelling of these regions), and much more.
----- Transcript of session follows ----- 421 nw.oirtorm.net.kiae.su (tcpld)... Deferred: Connection timed out during user open with oirtorm.net.kiae.su
----- Unsent message follows ----- Received: from anlemc.msd.anl.gov by cpuv1.net.kiae.su with SMTP id AA11761 (5.65.kiae-1 for {alekseev-at-nw.oirtorm.net.kiae.su} ); Fri, 25 Feb 1994 02:43:27 +0300
In response to requests for suppliers of protective lacquer for electropolishing I would like to inform you that Microshield Lacquer is available from:
South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 USA
The Microshield Lacquer has been used for many years with our Model 550 Electropolisher and Model 550 Chemical Jet Polisher. It is acetone soluble and is available in an 8 ounce kit. Please reference P/N 02-01890-01.
} } Is there anyone who is doing this or has experience with muscle firber analysis } } that can recommend software for either the PC or Mac platform? } } NIH-Image, a freeware image processing program for the Macintosh } available from the National Institutes of Health at zippy.nimh.nih.gov, } directory /pub/image, has facilities for image alignment (in several } modes), area determination (including thresholding/density slicing to } segment the regions of interest, and automatic counting, analysis and } labelling of these regions), and much more.
Here's the README file from Image:
NIH Image --------- NIH Image is a public domain program for the Macintosh for doing digital image processing and analysis. It can acquire, display, edit, enhance, analyze, print, and animate grayscale and color images. It reads and writes TIFF, PICT, PICS and MacPaint files It features multiple windows, MacPaint-like editing and 8 levels of magnification. It supports Data Translation and Scion frame grabber cards. Image requires at least 4MB of RAM and 8-bit video. The following files in the directory /pub/image contain NIH Image, documentation, source code, and example images.
nih-image1xx_fpu.hqx NIH Image 1.xx application. Requires FPU. nih-image1xx_nonfpu.hqx Version that does not require floating-point nih-image1xx_docs.hqx Documentation in Word 5.0 format nih-image1xx_source.hqx Think Pascal 4.0 source images Directory with images in TIFF and PICT format stacks Example stacks(3D images and movies) (directory) nih-image_spinoffs Contains variants of NIH Image(directory) programs Directory containing miscellaneous related programs
File Formats ------------ Files are in one of three formats. Those with a ".hqx" suffix are BinHex encoded Mac binary files, those with a ".bin" suffix are MacBinary encoded Mac binary files, and those with a ".txt" suffix are a plain text files. The BinHex and MacBinary formats represent the two parts of a Mac file(the data fork and the resource fork) as a single file. They permit storage of a complete Mac file on a non-Mac system, such as this server.
Most files were compressed using the Mac utility Stuffit 1.5.1 and uploaded using Fetch, which does the BinHex or MacBinary encoding. Both utilities can be found in the util directory. The best way to retrieve files is to use Fetch, which automatically does Binhex (or MacBinary) decoding and file decompression. Unfortunately, Fetch requires a Mac directly connected to the Internet. If this is not the case, use an FTP (File Transfer Protocol) utility to transfer the files to a local host and then transfer them to a Mac via modem.
For Macs not directly on the Internet, BinHex files must be transferred to a Mac using "ascii" mode and then decoded and decompressed using Stuffit or some other Mac compression utility, such as Compact Pro. MacBinary files must be transferred to an intermediate host using FTP in "binary" mode, then transferred to a Mac in "MacBinary" mode and decompressed using Stuffit or Compact Pro. A copy of Stuffit 1.5.1 is in the directory /pub/util in MacBinary format. The document "ftp-primer.txt" in the documents directory provides more information on file formats and FTP.
NIH Image Mailing List ---------------------- There is an NIH Image mailing list. It was set up by a group in the Soil Science Department at the University of Minnesota. To subscribe, send a message containing the line "subscribe nih-image {your name} " to soils.umn.edu.
DepthGauge ---------- DepthGauge is a control panel that allows rapid switching between monitor depths settings(e.g. 8-bit color and 24-bit color).
Fetch(/pub/util) ---------------- Fetch is a slick utility that allows networked Macs to transfer files over the Internet using the File Transfer Protocol(FTP). It does BinHex decoding and file decompression as the files are transferred.
Giffer(/pub/image/peograms) ----------------------- Giffer is a shareware program useful for converting from GIF to Pict format, and vis-versa.
ImageFFT(/pub/image/image_spinoffs) ----------------------------------- ImageFFT is an extension to NIH Image to support frequency domain (power spectrum) display and editing. It can do a 512x512 FFT in 18 seconds on a Mac IIfx.
ImageFractal(/pub/image/image_spinoffs) --------------------------------------- ImageFractal is a version of Image modified to compute the Fractal Index of objects by the Richardson Plot or the Tile-amalgamation methods.
Image/MG(/pub/image/image_spinoffs) ----------------------------------- Image/MG is an extension of Image supporting quantitative evaluation of cerebral blood flow, glucose metabolism, and protein synthesis.
NCSA PalEdit(/pub/image/programs) ----------------------------- NCSA PalEdit is a public domain program from the National Center for Supercomputing Applications for creating and customizing color palettes. With PalEdit, you can modify the whole palette or individual entries in the palette, to create a set of colors tailored to your needs. PalEdit can save palettes in a format compatible with NIH Image.
MockWrite --------- MockWrite is a simple DA text editor that is convenient for editing macros written in Image's Pascal-like macro programming language.
Projectionist(/pub/image/programs) ------------------------------ Projectionist is a utility that allows you to animate stacks created by Image and saved in PICS format. Because all the frames in the stack do not need to be loaded into RAM, Projectionist requires less memory than image. This preliminary version uses the standard system palette, so stacks created with Image may lose some of their colors when animated with Projectionist.
There are a couple of solutions to this problem that we've used. First, capture one image, then print it on mylar with a laser printer (or to paper and photocopy onto mylar if your printer doesn't do mylar). then capture the image to be overlaid and hold the mylar transparency over the computer monitor. This usually works best with a high contrast image on the mylar- otherwise the gray in the background details will obscure the lookup image on the monitor. this works well for physical disectors. The second image can be grabbed during the long print time.
Second, NIH Image, a public domain image analysis program for the Mac, allows alignment of two overlaid images. You can capture the first image, duplicate it, and then capture the second image and perform a live paste which superimposes the live image over the captured image. Live paste allows moving the stage until the live image aligns. - This sounds easy, but usually quite tedious in practice.
The simplest way to align images in Image is to capture your images and then run one of the alignment macros that either come with Image or are avaialble from the User_macros folder from Zippy.nimh.nih.gove (NIH Image ftp source). One alignment macro allows drawing lines on images in a stack, then all members of the stack are automatically aligned. Another macro, which I wrote, works on pairs of images. Draw a line between two landmarks common to the two images, then the macro will rotate one of the images to rotationally align the two lines and transparently paste it over the other image. You tweak the XY registration by hand.
Most PC software with macro capabiblity should allow creating the same type of macros. 24 bit software may allow putting the two images to be aligned into different color planes, then aligning. - Glen MacDonald Hearing Development Laboratories RL-30 University of Washington Seattle, WA 98195 (206)543-8360 glenmac-at-u.washington.edu -
We just tried Evan's blue for the same purpose. It was somewhat visible in wet tissue coverslipped with glycerol. Dehydrating through ethanols and clearing in xylene, coverslipping with DPX (our standard protocol for paraffin and vibratome sections) dramatically improved the fluorescence. Nearly one week later there is no noticeable fading or diffusion. Sure is pretty.
Could you email your labelling method - concentrations, route of administration and survival times? I'll get the grad. student who did this to make her method available to send to you.
Glen MacDonald Hearing Development Laboratories RL-30 University of Washington Seattle, WA 98195 (206)543-8360 glenmac-at-u.washington.edu
I have read on this e-mail system about polymerization of LR WHITE RESIN in a vacuum oven. Do you close the lids of the BEEM capsules when in the oven or leave them open?
I`m interested in the morphology of Panthera sp. epidermis. If anyone knows something about it, please, contact me. Thank you.
============================================================================= Francisco Javier Hernandez Blazquez | Universidade de Sao Paulo - Faculdade de | e-mail fjhblazq-at-fox.cce.usp.br Zootecnia e Engenharia de Alimentos | Voice: 55 195 616122 r. 265 Departamento de Ciencias Basicas/Histologia| Av. Duque de Caxias Norte, 225 CP 23 | Fax: 55 195 611689 CEP 13630-000 Pirassununga (Sao Paulo) | BRAZIL | ==============================================================================
I`m interested in the morphology of Panthera sp. epidermis. If anyone knows something about it, please, contact me. Thank you.
============================================================================= Francisco Javier Hernandez Blazquez | Universidade de Sao Paulo - Faculdade de | e-mail fjhblazq-at-fox.cce.usp.br Zootecnia e Engenharia de Alimentos | Voice: 55 195 616122 r. 265 Departamento de Ciencias Basicas/Histologia| Av. Duque de Caxias Norte, 225 CP 23 | Fax: 55 195 611689 CEP 13630-000 Pirassununga (Sao Paulo) | BRAZIL | ==============================================================================
I have obtained fixed (2.5% glutaraldehyde in 0.1M sodium cacodylate buffer) lung (rabbit) tissue which has been embedded in LR White and Spurr resin. However, they have NOT been stained with osmium tetroxide prior to embedding. Could anyone suggest possible methods of osmium staining post sectioning? Thankyou.
Microscopists: I am interested in imaging from my TEM directly. For starters, let's say at 1Kx1Kx8bit. Is anyone out there putting the moral equivalent of a CCD in the column? (I suspect that you'd cook an actual CCD in a hurry, not to mention that you don't need response to photons). I see problems with file size, but what are the physical reasons that we don't capture images this way? Julian Smith III Dept. of Biology Winthrop University Rock Hill, SC 29733 Vox 803-323-2111 Fax 803-323-2246
subject:-TEM-PREPARATION OF TEM SAMPLES(PURE COPPER)
I would want to know if it is possible to indicate and to know the initial macro shear direction on a photo of a 3mm TEM disk sample submitted to an shear deformation by extrusion.Moreover, can we prepare and observe square samples instead of conventional disk samples for TEM.I know that some studies have been made in Russia using square samples and showing the direction of initial shear on the pictures. Thanks a lot by advance for any help. Stephane Ferrasse Email address:S0F6296-at-ZEUS.TAMU.EDU
I am studying pure copper samples with the Phillips EM400 and I have problems because my samples seem to be too thick to observe grains,subgrain boundaries and dislocations that I want to observe. My preparation is the following: 1-rough polishing with 600 grit abrasive powder to produce 100-150 micrometers thick parallel sided sheets by using a simple jig. 2-use of a conventional punch device to obtain a 3mm disk 3-use of the Tenupol automatic double jet electropolisher The electrolyte is:20% nitric acid-80% methanol (conditions:methanol cooled in dry ice). QUESTIONS: 1-I don't know if the voltage that I use is good(8-10V);the same for the value of my flow rate (it takes me about 2-4mn to thin my specimen under these conditions) 2-Is it better to have small (those I obtained are about 40-100 micrometer in diameter) or big holes for TEM observations ? 3-For observing grains and subgrains do we have to use diffraction patterns or Kikuchi lines (as it is the case for dislocation observations) to find a better orientation ? More generally, is orientation important to see grain boundaries? 4-I use the highest voltage (120 kV) .Is it correct? 5-Is there something wrong in my initial preparation ( electrolyte,..)? Thanks a lot by advance for any help, Yours Faithfully. Stephane Ferrasse University of Texas A&M phone:409-845-1790 (USA) Email address:S0F6296-at-ZEUS.TAMU.EDU
Message-ID: {MAILQUEUE-101.940307083235.256-at-anat.uct.ac.za} To: microscopy-at-anlemc.msd.anl.gov
I am passing on a query from Peter Linder (Linder-at-botany.uct.ac.za) regarding clearing agents for Botanical Specimens.
He is working with pollen and was wondering if anyone knew of a clearing agent(s) that dealt effectively with tannins. The ones he has used tend to make a mess of the tannins so that they form obnoxious lumps.
Any suggestions?Peter D. G. Richards Dept Anatomy and Cell Biology UCT Medical School Observatory 7925 RSA Tel: 021-406 6285.
Does anyone know the manufacturer of gelatin capsules or if a conical gelatin capsule is available? Since LR-White doesn't seem to work with BEEM capsules, has anyone tried UniCryl with BEEM capsules? I'm doing a lot of cell suspensions for immuno and a conical capsule is ideal. I appreciate all info on this subject. Thanks, Phil Rutledge prutle1-at-gl.umbc.edu
Sponsored by Japan Chapter of SPIE, Co-sponsored by BACUS and SPIE
22 April 1994 Kanagawa Science Park Kawasaki City, Kanagawa Japan ============================================================
Contents ========
1.0 Symposium on Photomask and X-Ray Mask Technology 2.0 Hotel Accommodations 3.0 Registration Information 4.0 General Information 5.0 How to Receive More Information
1.0 PHOTOMASK AND X-RAY MASK TECHNOLOGY =======================================
Symposium Chair: Touru Yoshizawa, SPIE Japan Chapter Steering Committee Chair: Yoshio Tanaka, Oki Electric Industry Co., Ltd. (Japan)
Steering Committee Members: Naoaki Aizaki, NEC Corp. (Japan); Hideaki Hamada, ETEC Systems Japan Ltd. (Japan); Naoya Hayashi, Dai Nippon Printing Co., Ltd. (Japan); M. Ohtaki, Toppan Printing Co., Ltd. (Japan); K. Nakashima, Lasertec Corp. (Japan); Norio Saitou, Hitachi, Ltd. (Japan); Kazumasa Shigematsu, Fujitsu Ltd. (Japan); Yoshiki Suzuki, KLA Japan Ltd.; Yoichi Takehana, HOYA Corp. (Japan); Tadahiro Takigawa, Toshiba Corp. (Japan); Yoshihiro Todokoro, Matsushita Electronics Corp. (Japan); Yaichirou Watakabe, Mitsubishi Electric Corp. (Japan); Hideo Yoshihara, NTT Advanced Technology Corp. (Japan); James A. Reynolds, Reynolds Consulting (U.S.)
2.0 HOTEL ACCOMMODATIONS ========================
The Japan Travel Bureau, Inc. (JTB) has been appointed as the official travel agent for the symposium and will handle reservations of hotel accommodations. Inquiries and applications should be addressed to:
Japan Travel Bureau, Inc. International Travel Division Convention Center (Ref. CD100748-014) 1-13-1 Nihombashi, Chuo-ku, Tokyo 103 Japan
Hotel KSP (Symposium site) 3-2-1 Sakado, Takatsu-ku Kawasaki, Kanagawa 213 Phone: +81-44-819-2211 Y12,650 single w/bath Y24,200 single use of twin room Y25,300 twin w/bath
Hotel Sunroute Den-En Takatsu (15-minute walk to the conference site) 508 Futako, Takatsu-ku Kawasaki, Kanagawa 213 Phone: +81-44-814-3122 Y8,500 single w/bath Y18,000 twin w/bath
Note:
1. The room rates do not include meals. 2. 3 or 6% tax and 10% service charge will be added to the bill when checking out. 3. Should you fail to arrive at the hotel on your scheduled check-in date, your reservation will be automatically canceled.
Application and Payment -----------------------
Participants wishing to reserve hotel accommodations should contact the JTB no later than 20 March 1994. Application for hotel accommodations should be accompanied by remittance of the hotel deposit (one-night room charge) plus a handling fee of Y500 and sent to JTB. No reservation will be confirmed in the absence of this payment. Personal checks are not accepted. All payments must be in Japanese yen. Payment should be in the form of:
* a bank transfer to the Japan Travel Bureau, Inc., account at the Bank of Tokyo, Marunouchi Office, 1-4-2 Marunouchi, Chiyoda-ku, Tokyo 100 Japan (Account number 1025740/Ref. CD100748-014)
* bank draft payable to the order of the Japan Travel Bureau, Inc.
* The following credit cards may be used for payment of hotel deposit: Master Card, Diners Club, Visa Card, AMEX.
Cancellation ------------
In the event of hotel reservations which must be canceled, written notification should be sent to JTB. The following cancellation fees will be deducted before refunding:
Up to 9 days before the first night of stay: Y2,000 Up to 2 to 8 days before: 20% of daily room charge (minimum Y2,000) Less than 2 days before, or no notice given: 100% of daily room charge
3.0 REGISTRATION INFORMATION ============================
To preregister for Photomask Japan '94 contact:
Secretariat-Photomask Japan '94 c/o Business Center for Academic Societies Japan Conference Department 5-16-9 Honkomagome, Bunkyo-ku, Tokyo 113 Japan
Phone: +81-3-5814-5800 Fax: +81-3-5814-5823
* Registration Fees:
SPIE Member . . . . . . . . . .Y25,500 Working Group Member (e.g., BACUS) . . . . . . . Y28,500 Nonmember . . . . . . . . . . .Y30,000
* Cancellation Policy:
No refunds will be made for cancellations received after 16 April 1994.
* Method of Payment (choose one):
All payment must be made in Japanese yen.
* A bank draft for total fee payable to the order of Photomask Japan.
* Bank transfer to the account of Photomask Japan, A/C No. 075-1834926, Daiichi Kangyo Bank, Hongo Branch, Tokyo.
* Credit card (Visa, Diners Club, MasterCard, or American Express)
On-site registration will be accepted at the conference site on 22 April. If you must register on site, please plan to arrive 30 minutes before the program begins. Registration badges are required for admittance to the conference. The technical conference will begin at 8.30.
4.0 GENERAL INFORMATION =======================
* Registration and Information Hours
Registration takes place in front of KSP Hall, Third Floor
Friday 22 April 8.00 to 16.00
* Speakers'/Chairs' Registration Desk
Located in front of KSP Hall, Third Floor
Friday 22 April 8.00 to 16.00
* Location/Travel Information
Photomask Japan '94 will be held at the Conference Hall of Kanagawa Science Park (KSP Hall), 3-2-1 Sakado, Takatsu-ku, Kawasaki, Kanagawa, Japan. Phone: +81-44-819-2211
* Climate
The temperature in April ranges between 10 degrees C and 19 degrees C and the average humidity is 65%.
* Visa Requirements
Participants from countries which require a visa to enter Japan should apply at the nearest Japanese embassy well in advance of departure. If you have any questions or requests to obtain a visa, please contact the Secretariat-Photomask Japan ;94 as soon as possible.
Secretariat-Photomask Japan '94 c/o Business Center for Academic Societies Japan Conference Department 5-16-9 Honkomagome, Bunkyo-ku, Tokyo 113 Japan
Phone: +81-3-5814-5800 Fax: +81-3-5814-5823
5.0 HOW TO RECEIVE MORE INFORMATION ===================================
The complete text of the printed advance technical program for Photomask Japan '94 is available via anonymous FTP at:
It is also available through SPIE's automated e-mail server:
Send an e-mail message to,
info-optolink-request-at-mom.spie.org
with the following text in the message body:
send [optolink.meetings.programs]FILENAME.txt
To request a printed advance technical program via e-mail contact:
spie-at-mom.spie.org
For information regarding this meeting or other SPIE symposia or publications, contact SPIE at:
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This advance technical program is based on commitments received up to the time of publication and is subject to change without notice.
---------------------------------------------------------------- SPIE is a nonprofit society dedicated to advancing engineering and scientific applications of optical, electro-optical, and optoelectronic instrumentation, systems and technology. Its members are scientists, engineers, and users interested in the reduction to practice of these technologies. SPIE provides the means for communicating new developments and applications to the scientific, engineering, and user communities through its publications, symposia, and short courses.
SPIE is dedicated to bringing you quality electronic media and online services.
********************************************************************** Biological X-ray microanalysis - applications and recent developments **********************************************************************
The biological XRMA group of the Royal Microscopical Society are holding their Spring Meeting at the University of Sheffield on April 7th 1994. Papers will be presented on a variety of subjects including specimen preparation, elemental mapping, analysis of plant tissue extracts and biological-material interactions. Registration fees are 25 pounds for RMS members and 35 pounds for others.
Further details are available from:
Dr Paul Hatton, School of Clinical Dentistry, University of Sheffield, Claremont Crescent, Sheffield S10 2TA, UK
Sender: kayton-at-ohsu.edu (Robert Kayton) Message-Version: 2 } To: microscopy-at-anlemc.msd.anl.gov
We just tried Evan's blue for the same purpose. It was somewhat visible in wet tissue coverslipped with glycerol. Dehydrating through ethanols and clearing in xylene, coverslipping with DPX (our standard protocol for paraffin and vibratome sections) dramatically improved the fluorescence. Nearly one week later there is no noticeable fading or diffusion. Sure is pretty.
Could you email your labelling method - concentrations, route of administration and survival times? I'll get the grad. student who did this to make her method available to send to you.
Glen MacDonald Hearing Development Laboratories RL-30 University of Washington Seattle, WA 98195 (206)543-8360 glenmac-at-u.washington.edu
There is an immediate opening for a Scanning Electron Microscopy (SEM) Technician at Exxon Research and Engineering Company's Corporate Research Laboratory in Clinton, New Jersey. As a member of the Microcharacterization Team at Corporate Research, this position will involve the operation of a JEOL SEM with associated Energy Dispersive and Wavelength Dispersive Spectrometers. The position will involve the creative application of high resolution SEM imaging and elemental characterization of samples related to a wide range of projects at Exxon's Corporate Research Laboratory. The position will also involve general laboratory operations, sample preparation, SEM maintenance and computer analysis of the SEM data (both image analysis and spectral analysis).
Successful candidates should have experience in chemistry, physics or material science with a Baccalaureate degree or equivalent experience. Previous SEM experience and experience with high vacuum systems and computers (DOS, Unix, and VMS) is highly desirable. Since a number of projects are simultaneously in progress, it is essential for the SEM technician to be very organized and to pay attention to detail and accuracy in reporting results.
Qualified candidates please send resume to:
William Lamberti Associate Research Physicist Exxon Research and Engineering Company Clinton Township Route 22 East Annandale, New Jersey 08801
Equal Opportunity Employer M/F/H/V
All resumes must be received by April 8, 1994.
William A. Lamberti Office LA-196 Exxon Research and Engineering Company Route 22 East Annandale, NJ 08801 Email: walambe-at-erenj.com
TEM-PREPARATION OF PURE COPPER SAMPLES Thank you first to all those who have given me useful information about preparation of pure copper samples. I would want to know however something else about specimen washing and more precisely about storage of copper.I have read (Eddington-Practical electron microscopy) that Cu can not be stored because of oxydation.Then must we use Cu immediatly after jet polishing or can we keep it in a dessicator?If it is the case, for how long? Thanks in advance for any help. Regards. Stephane Ferrasse University of Texas A&M--email:S0F6296&ZEUS.TAMU.EDU
I realize that the Microscopy Listserv is predominantly used for those microscopies involving light or electrons, however, for those interested in "ions" this may be of interest!
(page 1)
- FIRST NOTICE -
SEVENTH ANNUAL WORKSHOP ON SECONDARY ION MASS SPECTROMETRY
Microelectronics Center of North Carolina (MCNC) and Holiday Inn - RTP
Research Triangle Park, NC
May 10-12, 1994
This workshop is an informal gathering of scientists for discussions relating to fundamental aspects, instrumentation and applications of Secondary Ion Mass Spectrometry.
PROGRAM
On Tuesday evening, May 10, 1994, the Seventh Annual Workshop on SIMS will commence with a registration/mixer from 7 - 9 pm. On Wednesday morning, May 11, a special topics session will focus on fundamentals, application and techniques related to secondary ion imaging. Other topics of interest will be accepted for inclusion in follow-on sessions or the third day's program. Following Wednesday's sessions there will be an evening cocktail hour, dinner, and vendor presentations. The third day will be devoted to user's group meetings and contributed presentations.
The format for this workshop will be informal, with topics of interest to both the novice and experienced SIMS user. You are encouraged to prepare an oral or poster presentation on any aspect of SIMS that may be of interest to your fellow workshop attendees. Open forum user's groups are planned for magnetic sector, time-of-flight, and quadrupole-based mass spectrometer instruments. Technical representatives from the instrumentation companies have been invited to address issues involving instrumentation, modifications, methodologies, and new equipment.
______________________ (page 2)
PRESENTATIONS Please indicate on the registration form provided if you are interested in presenting an oral or poster presentation on a topic which may be of interest to other workshop attendees. An abstract must be submitted to the organizing committee by April 15, 1994. As a special option this year, you are invited to submit a full length manuscript for a special issue of the new journal - Microbeam Analysis .
REGISTRATION Please note: the registration fee is $50.00 if received before April 25, 1994; a late registration fee of $100 will apply after this date. The SIMS Workshop is now being held under the auspices of the Microbeam Analysis Society. Checks should be made payable to: Microbeam Analysis Society and sent directly to Dr. Steven Hues with the completed registration form below - to be received no later than April 25, 1994. Note: The registration fee is waved for full-time student presenters only!
LOCAL ACCOMMODATIONS A block of rooms has been reserved at the Holiday Inn - Research Triangle Park, NC [919-941-6000], at a reduced rate of $72/night + tax (mention the SIMS Workshop). Rooms must be reserved before April 19, 1994 to be eligible for this reduced rate. All inquires pertaining to reserving rooms or details on the facilities, as well as payment for your room, should be made directly to the Holiday Inn - Research Triangle Park, NC. The closest airport with free Holiday Inn Shuttle connection (~10 min.) is the Raleigh-Durham International. Bus transportation to and from the hotel and MCNC is also provided.
____ Yes, I will submit a manuscript for publication in a special issue of Microbeam Analysis.
_______________________ (page 3)
ORGANIZING COMMITTEE:
Steven M. Hues Naval Research Laboratory (202) 767-2671
Greg Gillen National Institute of Standards and Technology (301) 975-2190
Richard T. Lareau Army Research Laboratory (908) 532-0119
IMPORTANT: Registration form and the $50 registration fee must be received no later than April 25, 1994 !!!
Please complete the registration form provided (see pg. 2), cut at dotted line, and return with registration fee enclosed (made payable to the Microbeam Analysis Society) to:
Steven M. Hues Naval Research Laboratory Code 6170 4555 Overlook Ave., S.W. Washington, D. C. 20375-5342
____________
Best regards,
Richard T. Lareau, Ph.D. US Army Research Laboratory Electronics and Power Sources Directorate AMSRL-EP-EC-M, Myer Research Center Fort Monmouth, New Jersey 07703-5601
We are considering the purchase of a stage heater/cooler for the Nikon Diaphot. Our main concern is cooling, e.g. stable tempertaure at approx. 22 degrees C. Does anybody have a recommendation of a particularly good stage temperature controller. Right now we have a water cooled system but problems include 1.) vibration and 2.) too narrow for access with high N.A. objectives. Thanks- Michael Cammer
Message-Id: {MAILQUEUE-101.940310095650.288-at-redhot.hut.fi} To: MICROSCOPY-at-ANLEMC.MSD.ANL.GOV
The European Microbeam Analysis Society (EMAS) organises
The 1st EMAS Regional Workshop on June 15-17, 1994, in Helsinki, Finland:
Electron probe microanalysis of materials today - practical aspects
The workshop is intended as a comprehensive introduction for those who are involved with SEM+EDS or EPMA based microanalysis applied to inorganic materials. The emphasis will be on the possibilities and limitations of the methods, and to give enough guidelines, procedures and examples in order to keep the discussion, for which there will be ample time, on a practical and basic level. The participants are encouraged to bring in their own problems for debate. The workshop language is English.
Main subjects:
* Electron-specimen interaction: X-ray generation & detection, spatial resolution * Analytical electron microscopy (AEM): possibilities and limitations * Correction procedures in microanalysis * Practical problems with EDS analysis & practical aspects of WDS analysis * Characterisation of EDS detectors * Standardless vs. quantitative EDS analysis: what can you expect? * SEM + EDS or EPMA with WDS? * Thin surface layer (1 nm - 1 m) analysis with EPMA or SEM + EDS * AEM as a tool for practical problems * How accurate are your results? * Strategy for applying microanalytical techniques
The main lecturers are: Dr. Peter Karduck (Gemeinschaftslabor fuer Elektronenmikroskopie, RWTH Aachen), Dr. W. A. Patrick Nicholson, Dept. of Physics & Astronomy, Univ. of Glasgow) and Dr. Peter Willich (Fraunhofer Institut fuer Schicht- und Oberflaechentechnik, Hamburg).
Posters are invited. Interpretation of the word "regional" can be done in a broad sense. For additional information please see below.
Erkki Heikinheimo ************************************************************* Erkki Heikinheimo e-mail: eheikin-at-redhot.hut.fi Helsinki Univ. of Technology Lab. of Metallurgy SF-02150 Espoo phone +358-0-4512759 FINLAND fax +358-0-4512798 *************************************************************
Has anyone had any experience in processing bacteria grown on 3mm glass beads for SEM? The cells are grown in suspension and the positioning of the beads is irrelevant. Want to look at surface for adhesion properties study. I want to try normal SEM processing procedures but would appreciate any info on an easy method. Thanks, Phil Rutledge prutle1-at-gl.umbc.edu
We are working on an immunoEM project localizing different epitopes in S. mansoni worms. Some of the life cycle stages are fairly small. We are embedding in Unicryl but we have some problems with the small specimens. I tried to put them in gelatine but this gives a problem with the sectioning properties of Unicryl. Does anyone have a better solution. We thought of putting them in agar, but I don't like the temperature this requires (immunoreactivity).
Greetings, We use LOW gelling temperature agar for our tiny specimens. Sigma sells a variety of these, and we use type VII. I forget exactly what the melting T is, but it melts readily on the low setting of a hot plate. We fix, rinse, and then put a few microliters (5-15?) of molten agar around the sample. It sets up "instantly". We use a 2% agar/water solution. Then we dehydrate and embedd as usual. We also throw in a bit of fast green at 100% ethanol, for ease of finding the samples. I have taken this stuff into several kinds of resin and never had any prboblems sectioning, light or em level. Also, we routinely do immunocytochem localizing microtubules, a fairly senstive antigen, so I don't the heat is a big problem. I hope this is useful for you. Please reply if you have further q's. Good Luck, Tobias Baskin
} Hi, } } We are working on an immunoEM project localizing different epitopes in S. } mansoni worms. Some of the life cycle stages are fairly small. We are } embedding in Unicryl but we have some problems with the small specimens. } I tried to put them in gelatine but this gives a problem with the } sectioning properties of Unicryl. Does anyone have a better solution. We } thought of putting them in agar, but I don't like the temperature this } requires (immunoreactivity). } } Any help would be appreciated. } } -------------------------------------------------------------------- } Universitaire Instelling Antwerpen (UIA) } Lab Pathology } 2610 Wilrijk } BELGIUM } } Tel: 32.3.820.25.34 } Fax: 32.3.820.22.48 } E-mail: anapat-at-uia.ac.be } ------------------------------------------------------------------- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - Tobias I. Baskin Baskin-at-biosci.mbp.missouri.edu University of Missouri * Division of Biological Sciences * 109 Tucker Hall Columbia, MO 65211 USA voice: 314-882-0173 fax: 314-882-0123 ___ ____ ^ ____ _____ / \ / / \ / \ / / | / / \ / / /___ / /__ /_____\ / /__ / / / \ ( / / / / \ \ / / /____ / \ \____/ /_____
Does anyone know a reference with a derivation of the brightness equation relating brightness to accelerating voltage? Shimoyama, etal, 1972, seem to pull it from the air, and then everyone else seems to just reference them. Shimoyama has B=IeV/pikT where I is emission current density, V is relativistic voltage, T is filament temp. This is stated to be the relativistically corrected expression of the Langmuir theoretical value. Langmuir's equation is I=I(0) (eV/kT) sin2(phi) where I is the max current density in a focused spot, I(0) is the current density at the cathode, T is temp, and phi is the half angle of focused spot.
So does anyone know of a derivation of the brightness vs voltage equation that can be believed??
Subject: Time:11:24 AM OFFICE MEMO Bright vs kV Date:3/11/94 Roseanne: I think you can find the matter of the variation of brightness and accelerating voltage treated in a very understandable manner in Hall's 2nd Ed of "Introd. to Electron Micros.". One of the best general explanations of the characteristics of self-biased guns with tungsten filaments is contained in Chs. VI and VIII of the book "The Electron Microscope" by M. E. Haine, InterSci Pubs. I ran off some calcs of the variation of brightness vs both temp and kV for an illustration in class one time - if you send me your FAX number, I'll be glad to send you a copy. BIGELOW-at-UMICH.EDU FAX: 313-763-4788
Subject: Time:11:37 AM OFFICE MEMO Add'n on Bright. vs kV Date:3/11/94 Sorry, I neglected to mention that Hall's discussion of brightness vs kV appears on p. 158.
I don't do EM, and I've never worked with worms, but I have embedded small pieces of rat spinal cord in agar for vibratome sectioning and it didn't hurt immunoreactivity any for a number of peptides, 5-HT, or tyrosine hydroxylase. The agar stays molten at about 50-55 ”C, and your tissue is exposed to that for only a very short time. It may even help to cool (not freeze!) your samples slightly before embedding - just a thought. All I did was to attach the blocked pieces to a glass coverslip with SuperGlue, and then squirt the molten agar over and around them. It really isn't embedding in the sense that I'm sure the agar doesn't permeate the tissue at all, but it gave sufficient support to allow sectioning in a consistent plane. You might also consider Low Melting Point agarose (I believe it's available from BRL?) which would keep temperature to a minimum, but it's expensive. Hope this helps...
David Morilak Dept Psychiatry Stanford University morilak-at-cmgm.stanford.edu
RE: Getting a small hole while electropolishing Another "trick"
Another method of setting the automatice cutoff for light sensor s in electropolishers is to temporarily replace your specimen with a conventional TEM aperture. Use an aperture with about a 10-20 micron size hole (it can be an old used one out of your scope) and then turn on the entire electropolishing unit (except the voltage) get the flow of liquid running and adjust the "sensitivity" until your instrument just detects the hole. This is a bit more systematic than just arbitrarily setting the sensor without a calibration point.
You could try embedding in alginate. This can be done at room temperature (or cooler). We have used a method based on Tamponnet et al, Stain Technology 1988, v 63, 155-158. for ultrastructure of free cells and protoplasts.
Mix the cells with a 1-2% sodium alginate solution in buffer (0.1M cacodylate in the original). Extrude the solution through a narrow pippet/syringe into a solution containing 50mM Calcium chloride (or let drops fall into this, or spread a thin film on a slide and dip into this) where the alginate coagulates and holds the cells together.
The only problem we have had is with different batchs of alginate. Some coagulate well and some fail to.
} Do you have } reason to think that the epidermis in the cat family is different in some } special way from the epidermis of other mammals?
I wish thank you for your answer. I will give some explanation. I was in my lab when a engineer asked to speak to me. He was very excited about something he had discovered about the way that the epithelial cells of Panthera pardus epidermis are disposed. He wanted some information or a slide (histological section) of this animal to confirm his ideas, but he did not explained what ideas or what was his intention. He alleged professional secret. That sounds strange, isn t? I am a public employee so I must aid people who ask me aid, they pay my salary. I did a search in Biological Abstract and found nothing. I will do a search in Zoological Abstract, maybe I will be luckier. Thank you.
============================================================================= Francisco Javier Hernandez Blazquez | Universidade de Sao Paulo - Faculdade de | e-mail fjhblazq-at-fox.cce.usp.br Zootecnia e Engenharia de Alimentos | Voice: 55 195 616122 r. 265 Departamento de Ciencias Basicas/Histologia| Av. Duque de Caxias Norte, 225 CP 23 | Fax: 55 195 611689 CEP 13630-000 Pirassununga (Sao Paulo) | BRAZIL | ==============================================================================
I need to convert TIFF files generated in a PC program using specification 5 to TIFF files following specification 6 for use on a Silicon Graphics machine that requires specification 6. Does anyone know of a program that can do this conversion? Thanks for any pointers.
-- ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ Nancy L Desmond, Ph.D. Department of Neurosurgery University of Virginia Health Sciences Center, Box 420 Charlottesville, VA 22908
Message-Id: {MAILQUEUE-101.940314085545.384-at-parmly1.parmly.luc.edu} To: microscopy-at-anlemc.msd.anl.gov
} From: rutledge phil {prutle1-at-gl.umbc.edu}
} Has anyone had any experience in processing bacteria grown on 3mm glass } beads for SEM? The cells are grown in suspension and the positioning of } the beads is irrelevant. Want to look at surface for adhesion properties } study. I want to try normal SEM processing procedures but would } appreciate any info on an easy method. } Thanks, } Phil Rutledge } prutle1-at-gl.umbc.edu } Probably the quickest way is to pour your bacteria/beads solution into a funnel with a 0.22 or 0.45 micron nucleur-pore membrane filter, then run your fixatives/ OsO4/ dehydration etc. solutions through the same funnel, maybe 5 min. each by volume sample. Dry by CPD or hexamethyldisilizane at 60 C or Peldri. Pour out onto stub with something sticky & conductive on it, like conductive tape or a THIN coat partially dried silver paste (not paint). Phil Oshel
} Do you have } reason to think that the epidermis in the cat family is different in some } special way from the epidermis of other mammals?
I wish thank you for your answer. I will give some explanation. I was in my lab when a engineer asked to speak to me. He was very excited about something he had discovered about the way that the epithelial cells of Panthera pardus epidermis are disposed. He wanted some information or a slide (histological section) of this animal to confirm his ideas, but he did not explained what ideas or what was his intention. He alleged professional secret. That sounds strange, isn t? I am a public employee so I must aid people who ask me aid, they pay my salary. I did a search in Biological Abstract and found nothing. I will do a search in Zoological Abstract, maybe I will be luckier. Thank you.
============================================================================= Francisco Javier Hernandez Blazquez | Universidade de Sao Paulo - Faculdade de | e-mail fjhblazq-at-fox.cce.usp.br Zootecnia e Engenharia de Alimentos | Voice: 55 195 616122 r. 265 Departamento de Ciencias Basicas/Histologia| Av. Duque de Caxias Norte, 225 CP 23 | Fax: 55 195 611689 CEP 13630-000 Pirassununga (Sao Paulo) | BRAZIL | ==============================================================================
Sharath Yes you did miss something. Look in the EMMPDL subdirectory called CBED. There is a program there by John Porter which plots Stereograhic projections &CBED patterns. By adjusting the convergence angle you will get a conventional SAED pattern. I'm not sure if the UMICH library mirror has the master copy, however I would be very suprized if it doesnot since John Mansfield is very careful on backups and copies. I've appended a copy of the program abstract to this message.
Nestor Zaluzec ANLEMC --------------- Title :STERPROJ_IBM_V4 Keywords :Stereographic Projection, CBED, SAD, TEM, AEM, HOLZ Computer :IBM PC/XT/AT or compatible Operating System :PC-DOS Programming Language :QUICKBASIC 4.5 Hardware Requirements :Hewlett Packard HP7470A Plotter required. Author(s) :John R. Porter Correspondence Address :Rockwell International Science Center, :Thousand Oaks, CA 91360. Abstract:
This QuickBASIC program plots stereographic projections, CBED and HOLZ line simulations, and performs axis/angle pair calculations. Output is to the screen or a Hewlett Packard HP 7470A Plotter. Stereographic projections are drawn to scale for subsequent manipulations with a standard 18cm. Wulff net and can be plotted for cubic, hexagonal, tetragonal and monoclinic crystal systems in any orientation. Plotted poles can be restricted to those allowed by structure factor considerations (for certain structures) or restricted to those in certain Laue zones. The program leads the user through menus to determine the structure and orientation for the projection, which is then plotted accordingly. Version 4.00 is significantly enhanced compared to earlier versions.
EDITORS NOTE: THIS IS A NEW UPDATED VERSION OF STERPROJ- IBM (FEB.1990) -----------------------------------------------------------------------------
Subject: Time:5:16 PM OFFICE MEMO Bright. & Acc. Voltg. Date:3/14/94 My previous comment in response to Roseann Csencsits did not did not actually answer her question concerning the origin of the equation commonly used for brightness vs acc. voltage, because Hall doesn't take into account relativistic effects. A better source is the book "Transmission Electron Microscopy" by Reimer (Springer-Verlag 1984). The underlying equation is Eq. 4.10 on p. 90, which Reimer derives in a fairly understandable manner. This equation can be recast in terms of accelerating voltage U (in Reimer's terminology) by substituting eU for E and mc^2 for Eo (see table 2.1, p. 21), whereupon the terms for the 'relativistic voltage' Ur= U + (e/2mc^2)U^2 can be formed, and the equation becomes b = j/pi + (j/pi)(e/kT)Ur. Assuming the (j/pi) term is small compared to the rest of the equation, and can be neglected, this reduces to the approximation Roseann asked about. For a good definition of the 'relativistic voltage' see p. 30 of Spence. The experimental measurement of brightness is discussed by Spence in Sect. 7.2. Reimer also discusses this the brightness equation in Sect. 2.1 of his book "Scanning Electron Microscopy".
This is my second attempt to get some feedback on my request to get off the microscopy list. I've also sent the proper (but nonfunctional) message to listserver. Still I am swamped with msgs from the list.
Please remove my name from the list, and please let me know if there's a problem. I'm not the only person with this problem - there are enoguh of us that we are considering forming a new list: those_who_can't_unsubscribe_from_the _microscopy_list :}
This is my second attempt to get some feedback on my request to get off the microscopy list. I've also sent the proper (but nonfunctional) message to listserver. Still I am deluged with msgs from the list.
Please remove my name from the list, and please let me know if there's a problem. I'm not the only person with this problem - there are enoguh of us that we are considering forming a new list: those_who_can't_unsubscribe_from_the _microscopy_list :}
To: WALCKSD:ml:wpafb, SCHELTFJ:ml:wpafb Subj: Thinning Cerium Oxide Orig-Author: {del-at-sol1.lrsm.upenn.edu (David E. Luzzi)}:ddn:wpafb ----------------------------------------------------------- We need to thin a single crystal of Cerium Oxide (CeO2) to electron transparency but are having major difficulties with brittleness. Does anyone have experience with this, or analogous materials? Thanks in advance.
David E. Luzzi Dept. of Materials Science University of Pennsylvania 3231 Walnut Street Philadelphia, PA 19104-6272
I am looking for two second-hand reflective optical microscopes. One should be something like Bausch&Lomb Stereo Zoom 7, the other should have well calibrated focal depth. Can anyone tell me where to look? Thanks.
Lumin Wang TEM Lab., Univ. of New Mexico Bitnet: zircon-at-unmb
Lumin Wang wrote: } I am looking for two second-hand reflective optical microscopes. One should } something like Bausch&Lomb Stereo Zoom 7, the other should have well } calibrated focal depth...
I have no information on used microscopes. Maybe a dealer could help. I am, however, involved in importing new microscopes of all types. In fact, we have a 10X-160X zoom that is priced very low.
There have been a few problems during the last fe months with users trying to unsubscribe from the list. For the most part the problems were related to how they tried to unsubscribe, the MOST common problem was related to mail forwarding! The basic problem was that the particuliar user would subscribe from one address then have that computer forward mail to a different second address on a differnet computer system.
Upon attempting to Unsubscribe, the user would supply his/her second address to the server, rather than the original subscription address. Since the "second" address was unknown to the server nothing happened and they continued to receive their mail! This has happend at least a dozen times in the last few months so please remember to Unsubscribe you must inform the server of your original subscription address, not the one to which you have your mail forwarded. This is particuliarly important for sites/users that heavily use aliases and forwarding, so try to give me a break and remember from whence you came!.
The other problem which requires less investigative work, but is still a headache is that you must unsubscribe to the Listserver not to Microscopy List. (See below)
Also I've had a few comments come back saying that people are not sure if their messages have been posted. If your message is posted you WILL receive a copy back of your original message. The most common problem here is some subscribers simply "REPLY" to a message. This is fine AS LONG AS your Email system recognizes that the message came from the MICROSCOPY-at-ANLEMC.MSD.ANL.GOV, then there is no problem Some Email systems reply to the originator of the message (i.e. the person who started the question/comment). If this is the case then the members of this listserver will not see your reply! Please check your Email header before you send out a REPLY and insure that the message is going to MICROSCOPY-at-ANLEMC.MSD.ANL.GOV. If you do not receive a copy of the message you posted, then it is 99+% likely that your message was not posted to the subscription list....
Just a reminder to Unsubscribe you should send a message to:
Listserver-at-anlemc.msd.anl.gov
and include the following line in your mail message
Unsubscribe Microscopy Username-at-HostAddress
where Username = User Name that you originally subscribed with HostAddress = Address of the host you originally subscribed from ====================================================================
Alot of traffic goes through the computer now adays, and I'm pretty much swamped with things to do.... Yes, the new Alpha system has finally arrived, however, haven't had time to setup the software so it will still be awhile before anything is upgraded. We are now approaching the 900 subscriber mark so business is booming, but the Gov. has cut back our funds & people.
The never-ending-battle continues.........
Nestor J. Zaluzec ANL EM Center & Proprietor of the "Hotel California" Microscopy Listserver :-)
On the question of brightness vs voltage, I REPLYed, which sent my response only to the sender. Others may be interested in Principles of Electron Optics by P.W. Hawkes and E. Kasper, Academic Press, a good two-volume set with more than I want to remember about electron microscopy.
The 46th Annual Meeting of the Scandinavian Society for Electron Microscopy, June 13-15, 1994, Kuopio, Finland
INVITATION AND CALL FOR CONTRIBUTIONS
SCIENTIFIC PROGRAM
The aim of the symposium is to focus interest broadly in the recent developments of microscopy, not only emphasizing the importance of electron microscopy but paying attention also to the development of e.g. confocal microscopy, immunocytochemistry, image processing and image analysis.
The scientic program is designed to have two plenary sessions and parallel sessions for biological and materials scientists. The following sessions are included in the program: - High resolution microscopy - Confocal microscopy - Image processing, analysis and quantitation (plenary) - Electron microscopy in molecular biology - Scanning probe microscopies (plenary) - Electron beam assisted microanalysis - Immuno electron microscopy - Environmental cell pathology of plants - Electron microscopy in clinical medicine - New techniques and products
Invited speakers are amongst others A. Bardal (Norway), C.-H. von Bonsdorff (Finland), F. Cuisinier (France), S. Enestrm (Sweden), H.E. Gaub (Germany), H. Gundersen (Denmark), M.S. Gunthardt-Goerg (Switzerland), T. Holopainen (Finland), S. Huttunen (Finland), L. Kanerva (Finland), I. Kottke (Germany), T. Lepist (Finland), A.B. Maunsbach (Denmark), M. Mrgelin (Sweden), J. Paranko (Finland), J.C. Russ (USA), M. Ruhle (Germany), E. Soini (Finland), P. Willich (Germany).
LANGUAGE
The congress language is English
ABSTRACTS
Papers are invited on all aspects of electron microscopy and related techniques. The extended abstracts will be published as a separate proceedings book of the meeting. The abstracts, including photos and references, may contain two pages. The first page should contain text only, the second page may be used for text, figures and tables. The abstract must neve exceed two pages. Use a word-processor with TIMES 12 font and italics for taxonomic terms. The text and figures must fit inside a rectangle measuring 160 mm x 240 mm (width x height). One camera-ready original with one photocopy should be sent before March 31, 1994 to: Dr. Raija Tammi, Department of Anatomy, University of Kuopio, P.O.Box 1627, FI-70211, Kuopio, Finland
POSTERS
At the poster exhibition, a short time will be reserved to each participant for oral presentation. An area of 1100 x 1300 mm (width x height) is allocated for the poster.
SOCIAL PROGRAM
A Get-Together Party will take place on June 12, at the Snellmania Building of the University of Kuopio. On June 13, we take off for a cruise on Lake Kallavesi and continue with the smoke sauna and supper at Jatkn- kmpp (Lumberjacks' Lodgings). On June 14, there is reception at the City Hall and after this the conference dinner at the Hotel Arctia
REGISTRATION
The registration fee is 800 FIM for members, 950 FIM for non-members (if you join the society you will get advantage of membreship immediately), and 650 FIM for students and technicians. The fee includes the Get- Together Party, daily lunches and coffees and one copy of the published abstracts. Please contact SCANDEM 94 Secretariat, c/o Finnish Medical Society Duodecim, Savilahdentie 6, FI-70210 Kuopio, Finland (fax. Int.+ 358-71-240361). Deadline is April 15, 1994. Surcharge for registration is 150 FIM.
DEADLINES
March 31, 1994 Submission of abstracts April 15, 1994 Registration
LOCATION
Kuopio is located in the centre of the Lake-district of Finland about 400 km northeast of Helsinki. Connections to Kuopio by air, train or bus are good. The venue site of the meeting is the Snellmania Building of the University of Kuopio close to the centre of Kuopio.
David, I also routinely make TEM samples from GaAs and InP, which are reasonably brittle (but not as bad as YBCO). I simply mount the samples in thermoplastic wax on a glass slide about 20 mm x 20mm and polish down to about 100 microns using the finest grit paper I can find - 'worn out' 1200 grit paper is fine. I then mount it in Lacomit on a PTFE or nylon stub and chemically polish it to transparency using Cl in methanol jet from a wash bottle chopped in half and held upside down in a clamp. I can make 30 samples in a morning and the kit costs virtually nothing to make! The only thing to be careful of is to make sure that the wax ('Crystalbond' in my case) lies all around the sample so that no corners stick out, and to go slowly. If your material is as bad as 1-2-3 superconductor, you may need to be a bit more high-tech; I haven't done it myself, but work done here used a Dimpler to very slowly polish down to about 5 microns before ion milling to transparency. The samples took about 2 days to make and when they broke (about 1 in 5 samples) it was heart rending. I can give you the Email address of the person who did this, if you want - he's moved on now.
Regards,
Richard Beanland,
Department of Materials Science and Engineering, University of Liverpool, P.O. Box 147, Liverpool L69, 3BX, England.
} From: Francisco Javier H Blazquez {fjhblazq-at-fox.cce.usp.br} } } Do you have } } reason to think that the epidermis in the cat family is different in some } } special way from the epidermis of other mammals? } Try doing a literature search of "Science" and "Nature" for the last 5 or so years, using "self-organizing" and "Turing" in your keywords along with "leopard" & "jaguar". The title may have been something along the lines of "How the Leopard got his spots". Also, it may have been in the research news section and not an article. There was a short piece (which I copied & can't find), about some group discovering that the spots on a leopard's (or jaguar's) where a self-organizing phenomenon similar to some chemical reactions (the ones that swirl in a petri-dish?); the principle involved had been proposed orginally by Alan Turing (of math & comupter fame). Your engineer may have stumbled across this. ? Then again, it may be more mundane, like something to do with intercellular junctions. Phil Oshel (312) 274-6348
I just subscribed, got my welcome to microscopy notice and then sent out 2 messages regarding LM on the same day (about a week ago). I never saw either one or heard any response - although I have been catching up on the EM discussion of thinning CeO, etc. Just checking did my messages get out? Is there somewhere I can browse old messages from the list - an ftp site?
P. Joyce
Peter Joyce GRA Materials Science, University of Texas Phone: (512) 471-5723
I have two questions. We have a Leica MeF3 inverted metallograph we just purchased about a year ago. The bulb in the main lamphouse burned out and it was not easy to replace. The manual calls it a 12V/100W LV halogen bulb. When removed all it says on it is Philips. I tried to get Leica to tell me what it's called and who to get it from apart from them (they only had one in stock in Houston) and they were no help at all - although they did try to help me, sort of. The supplier they gave me as their source didn't know anything about being an OEM soure for Leica or Reichert or whoever they are??? What I'm looking for is anyone who has been this or as similar cycle with Leica equipment, or the name of a GOOD specialty bulb store that's been tried and tested on microscope equipment. I tried one really poor one in FLA and another not so bad in NY with little success.
Second question is related to an earlier post of mine from a few months back. I inquired about the use of index matching fluid to help observe my specimens. The response raised more questios than answers, thanks just the same. Experiments have shown that witha thin layer of oil and no coverslip (coverslip helps keep my optics safe from contamination, esp. inverted scope!) the difference in my specimens is night and day. I get really good clarity. The facts are - I'm looking at rough composite prepreg (carbon fibers in polysulfone) and the surface is much too diffuse to see anything. I went to our Plant Biology dept. who gave samples of permount oil, Zeiss immersion oil, mineral oil, and clove oil and some coverslips. The coverslips had little or no effect on optical performance with the oils. Also the Zeiss immersion oil was not very fruitful. The other 3 oils, in particular the clove oil gave excellent clarity at about 200x in darkfield. I went back to the Plant Biology guys for data on the clove oil, specifically the refractive index - they couldn't help me. I called the chemical supplier Sigma Chemical they haven't done that test. I checked the library, there no such data on clove oil and no mention at all of permount oil or mineral oil in the Chemistry CRC and the like. Is there anyone out there using the immersion technique who can comment on the use of such oils and the source of such data? Anyone ever studied clove oil? We're also observing a strange side effect, when the oil is applied tiny white lines, (maybe cracks in the surface of the material) become visible. We're not sure if they're there before the application of the oil because before the oil is applied we can't see well enough. The available data wouldn't suggest this type of oil should be attacking the polysulfone. Perhaps this effect actually arises from the oil getting in pre-existing cracks and highlighting them. Any comments.
P. Joyce
Peter Joyce GRA Materials Science, University of Texas Phone: (512) 471-5723
} I have two questions. We have a Leica MeF3 inverted metallograph we just } purchased about a year ago. The bulb in the main lamphouse burned out and } it was not easy to replace. The manual calls it a 12V/100W LV halogen } bulb. When removed all it says on it is Philips. I tried to get Leica to } tell me what it's called and who to get it from apart from them (they only } had one in stock in Houston) and they were no help at all - although they } did try to help me, sort of. The supplier they gave me as their source } didn't know anything about being an OEM soure for Leica or Reichert or } whoever they are??? What I'm looking for is anyone who has been this or as } similar cycle with Leica equipment, or the name of a GOOD specialty bulb } store that's been tried and tested on microscope equipment. I tried one } really poor one in FLA and another not so bad in NY with little success.
I just happen to have my local independent light microscope service person in the lab today doing routine maintenance. He tells me that, unless you're using a specialty housing, you should do fine with any standard 12V 100W lamp of the ANSI code FCR. You can get these from OSRAM, Philips or Ushio. There should not be any real difference between them. This lamp is rated at 50 hours lamp life at 12V.
John chandler-at-lamar.ColoState.EDU Fort Collins, CO
There were a couple of questions concerning LM bulbs and where to purchase them. I quit going to the manufacturers of LMs unless there was no alternative. I have been buying my bulbs from Bulbman,Inc. in Reno, Nevada for over 10 years. They seem to be pretty knowledgable about the bulbs and they have always been able to provide the bulbs or tell me where they may be purchased other than the manufacturer. Their toll free number is 1-800-648-1163. They also take PO's for small amounts of bulbs. You don't have to order more than you need to get a good price if the bulb you want is available. Hope this info helps. Phil Rutledge
I am seeking contact addresses/faxes/e-mail for distributors and manufacturers of CCD video cameras for biological light microscopy (currently brightfield, darkfield, phase and DIC) to be used in conjunction with a VCR and Macintosh Quadra 650 and image grabbing and manipulating facilities.
A colleague in this department uses a Pulnix TM-765 for similar purposes and this camera would be ideal. I have faxed the President of Motion Analysis, Inc at Eugene, OR, which was the the agent when he purchased his camera a couple of years back, but have had no reply. Can anyone give me other contacts or agents for Pulnix, and/or information on other manufacturers/agents for similar systems. We need a camera control system also, and hoped to pay only in the $2500-3000 range.
Any information on Australian distributors would also be welcome.
Thanks
Nikki Watson
Dr N.A. Watson Department of Zoology University of New England Armidale, NSW, 2351 AUSTRALIA Fax: 067 711 869 Phone: 067 732181
For additional support you can cut the conical tip from a BEEM capsule, place your sample insidek (i did this for rat spinal cord) and fill the capsule with agar. Then take the cap off, supergl;ue the thing to the slide after trimming down the open end of the BEEM capsule to expose a couple of millimeters of sample.
a thin walled polyethylened vial cap also works, if you can find an appropriate size, and will be easier to retirm if you need to keep sectioning farther down.
Glen MacDonald Hearing Development Laboratories RL-30 University of Washington Seattle, WA 98195 (206)543-8360 glenmac-at-u.washington.edu
On Fri, 11 Mar 1994, David Morilak wrote:
} I don't do EM, and I've never worked with worms, but I have embedded small } pieces of rat spinal cord in agar for vibratome sectioning and it didn't hurt } immunoreactivity any for a number of peptides, 5-HT, or tyrosine hydroxylase. } The agar stays molten at about 50-55 !C, and your tissue is exposed to that } for only a very short time. It may even help to cool (not freeze!) your } samples slightly before embedding - just a thought. All I did was to attach } the blocked pieces to a glass coverslip with SuperGlue, and then squirt the } molten agar over and around them. It really isn't embedding in the sense that } I'm sure the agar doesn't permeate the tissue at all, but it gave sufficient } support to allow sectioning in a consistent plane. You might also consider Low } Melting Point agarose (I believe it's available from BRL?) which would keep } temperature to a minimum, but it's expensive. Hope this helps... } } David Morilak } Dept Psychiatry } Stanford University } morilak-at-cmgm.stanford.edu } } ------- }
I would suggest buying refractive oils from Cargille Labs in Cedar Grove New Jersey, 201-239-6633. They sell liquids certified for refractive index from about 1.25 to 2.0 (this is from memory, and therefore mostly suspect). I have used refractive index kits containing 80 or so liquids that cover a range, but if you are just looking for something for immersion microscope objectives that is more consistant than clove oil they can supply one quarter ounce bottles.
} I am seeking contact addresses/faxes/e-mail for distributors and } manufacturers of CCD video cameras for biological light microscopy } (currently brightfield, darkfield, phase and DIC) to be used in conjunction } with a VCR and Macintosh Quadra 650 and image grabbing and manipulating } facilities. } } A colleague in this department uses a Pulnix TM-765 for similar purposes } and this camera would be ideal. I have faxed the President of Motion } Analysis, Inc at Eugene, OR, which was the the agent when he purchased his } camera a couple of years back, but have had no reply. Can anyone give me } other contacts or agents for Pulnix, and/or information on other } manufacturers/agents for similar systems. We need a camera control system } also, and hoped to pay only in the $2500-3000 range. } } Any information on Australian distributors would also be welcome. } } Thanks } } Nikki Watson } } Dr N.A. Watson } Department of Zoology } University of New England } Armidale, NSW, 2351 } AUSTRALIA } Fax: 067 711 869 } Phone: 067 732181 } } (using 'Eudora' on a Macintosh)
I have looked around in my files and have a few suggestions and corrections.
In my lab we use a couple of Hamamatsu C2400 CCD-video cameras, one with an intensifier coupled to it. I could not find the price for just the CCD camera + controller, but you can contact
Robert A. Wick, Ph.D. Photonic Microscopy, Inc. 2625 Butterfield Road, 103 West Oakbrook, IL 60521 Tel 312-571-1241 Fax 312-571-1244
Other addresses+numbers (form Laser Focus World, Buyers' Guide, 1994)
PULNIX America, Inc. 1330 Orleans Dr. Sunnyvale, CA 94089
Tel 408-747-0300 Fax 408-747-0880 ___________________________________
Dage-MTI Inc 701 N. Roeske Ave. Michigan City, IN 46360
Tel 219-872-5514 Fax 219-872-5559 ___________________________________
I am studying pure copper and copper-niobium samples with SEM.I would want to observe microstructural morphology of the specimen.Is some specific preparation of the specimen needed (for example special cleaning,etching...)?I would want to avoid any kind of preparation involving high temperatures. Thanks in advance for any help. Stephane Ferrasse Texas A&M University_E-mail:S0F6296&ZEUS.TAMU.EDU
A laboratory in our department is seeking a buyer/barterer for an Axiomat with good Nomarski (the machine was used for microtubule motility assays etc.) and other optics. If you are interested, please send us a message. Thanks- Michael Cammer cammer-at-aecom.yu.edu
Greetings, Has anyone tried dehydration in acetonitrile instead of ethanol, as suggested by Edwards et al Micr. Res. Tech 21:39-50 (1992)??? Experience with plant tissue would be particularly relevant, but any comments will be welcomed.
In Canada we have a vendor that sells Sony CCD video cameras, the address is: Soquelec, 5757 Cavendish blvd, suite 101 Montreal, Quebec, Canada, H4W 2W8 Tel:514 482-6427 fax:514 482 1929
Hoping that it will be of some help. Diane Montpetit Food Research Center, Agriculture Canada Quebec, Canada.
In Canada we have a vendor selling Sony CCD cameras: Soquelec 5757 Cavendish blvd, suite 101, Montreal, Quebec, Canada, H4W 2W8 tel:514-482-6427 fax:514-482-1929 Hoping that it will be of some help Diane Montpetit Food Research Center, Agriculture Canada, Quebec, Canada
Introduction to the structure of computer networks Local networks and network protocols; Larger networks; The Internet; Network structure at Tulane Medical Center
Local network functions Controlling network connections with the Chooser; File sharing; E-mail; Networked scheduling; Lab system access; Remote access (connecting from home); Software tools: Alias Manager; Microsoft Mail; Eudora; Meeting Maker; Termy; Apple Remote Access; ARA Commander.
Using the Internet IP addresses and MacTCP; E-mail and internet addresses; Listserv groups; Telnet and File Transfer Protocol (FTP); Transfer formats: binhexing and compression; Special communication protocols: the World Wide Web and Gopher; USENET news groups; Internet resources in the biomedical sciences; Software tools: MacTCP, NCSA Telnet, Fetch, Versaterm Link, Binhex 4.0, Stuffit, Compactor, Mosaic, TurboGopher, ph.
Reference searching and management Using Grateful Med for Medline searching; Current Contents on diskette; Reference management with EndNote; Software tools: Grateful Med 2.0, EndNote Plus, Microsoft Word
Procedures for connecting to the network at Tulane
In Canada we have a vendor selling Sony CCd cameras: Soquelec 5757 CAVENDISH BLVD, SUITE 101, MONTREAL, QUEBEC, CANADA, H4W 2W8 TEL:514 482-6427 FAX:514 482-1929
HOPING THAT IT WILL BE SOME HELP,, dIANE MONTPETIT, FOOD RESEARCH CENTER, QUEBEC, CANADA ,MONTPETITD-at-QCRSSH.AGR.CA
I am looking for suggestions on thin sectioning polymeric materials (polypropylene/synthetic rubber) for TEM analysis. I am having difficulty transfering sections (~80 nm) from the knife to grids. Common problems with the sections are curling and flying away. I was wondering if anyone has had any success embedding these types of materials. *************************************** David Garrett "DGARRETT-at-GAB.UNT.EDU" University of North Texas Dept. Biological Sciences (817)565-3964 Fax (817)565-4136 ***************************************
Plastic sectioning I am looking for suggestions on thin sectioning polymeric materials (polypropylene/synthetic rubber) for TEM analysis. I am having difficulty transfering sections (~80 nm) from the knife to grids. Common problems with the sections are curling and flying away. I was wondering if anyone has had any success embedding these types of materials. *************************************** David Garrett "DGARRETT-at-GAB.UNT.EDU" University of North Texas Dept. Biological Sciences (817)565-3964 Fax (817)565-4136 ***************************************
If you are sectioning on a dry knife you might like to try the trick that Tokuyasu used for recovering cryosections. Pick them up on the surface of a drop of sucrose or other inert solution held in a small wire loop. The liquid will stop your problems with static and the surface tension should flatten out the sections. Once the sections are recovered, place the liquid drop onto a coated grid. The recovery solution can be removed by floating the grid specimen face down on drops of water. I have seen this method used successfully for rubberized samples in Arizona. It will not be of much use to you if you cannot wet your sample.
Message in response to questions asked by Stephane Ferrasse, Texas A&M.
I assume you are interested in the morphology and distribution of niobium precipitates within a copper matrix. I have worked on rapidly solidified Cu-Nb microcomposites at Iowa State a couple of years ago and have published a technique which has given me excellent results in the SEM. Detailed procedures may be found in ASTM STP 1165, Metallography: Past, Present and Future, George Vander Voort et al., ASTM, 1993, article written by myself and collegues from Iowa State and Penn State.
Briefly, the technique involves an attack-polish method to remove some of the cumatrix. After a standard grind polish finishing on 0.25um diamond with kerosene as the lubricant, the sample was first immersed in a 20%HF, 20%H2SO4, 20%HNO3, 40%H2O solution to remove the smeared Nb created during polishing. Then the samples are immersed in a 30%H3PO4, 15%CH3COOH, 10%HNO3, 45% Ethanol solution to remove some of the copper surrounding the Nb ppts. To aide in this process, this step was performed in an ultrasonic cleaner - promoted homogeneous removal of copper. Final rinsing was done in the ultrasonic cleaner using fresh, 111 trichlorotrifluoroethane. The copper removal and final cleaningstep was repeated as necessary until the desired results were obtained.
Kevin L. Zeik U.S.Steel Technical Center Monroeville, PA 15146 EMail_Zeik-at-USS.COM
To: MICROSCO:CENTRAL cc: DRStadden:R_D:Armstrong Subj: Image Archiving Forwarded: Message from DRStadden:R_D:Armstrong of 3-23-94 ------------------------------------------------------------------ SEE FOLLOWING
--------------------- Forwarded Message Body --------------------- To: MICROSCO:CENTRAL cc: DRStadden:R_D:Armstrong Subj: Image Archiving ------------------------------------------------------------------ I'm curious about whether anyone is routinely electronically archiving images from SEM or LM, and/or sending them over a PC network. If so, what sort of hardware is required; particularly, what is retrofittable to a digital imaging SEM or a PLM, and what does it cost? I have one quotation for an SEM-interfaceable unit that is around $20K.
Thanks for any leads,
Dave Stadden Testing and Analysis Lab Armstrong World Industries, Inc.
When grabbing the image with CCD camera I a often getting problem with the very high signal which when looking at preview is almost white. I use f=22 but the signal is still to strong. The only solution besides a pice of hardware is to deminish the light source but it is a bad solution. Is there any software for Mac which can solve my problem ?
Michel Deschuyteneer suggested CD-ROM as a method for archiving images. This is something we have been trying to do for the last few months. If you plan to purchase a CD writer, you need to be aware that you may also need to buy a high speed hard disk from which the images will be downloaded to the CD. Look carefully at the input specifications of the CD writer before you buy the high speed disk.
Russell E. Cook Electron Microscopy Center for Materials Research Materials Science Division Argonne National Laboratory Argonne, Illinois 60439 USA
Russell E. Cook Electron Microscopy Center for Materials Research Materials Science Division Argonne National Laboratory Argonne, Illinois 60439 USA
------------------ RFC822 Header Follows ------------------ Received: by qmgate.anl.gov with SMTP;25 Mar 1994 09:41:29 U Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Ron Wroblewski asked about software for the Mac to help his intensity saturation problem....
The simple answer is you need a hardware fix not a software one. If the CCD is saturating then there is no software solution. You must decrease the signal. Some CCD camera's have autoIris lenses which attempt to adjust the input to the CCD by closing down the iris via a motorized mechanism.. Alternatively you could put in some type of neutral density filter to reduce the signal that is hitting your CCD.
Yes, we have just (today!) purchased the above printer for producing STEM/XRAY images primarily. For effectiveness/cost ratio it is nothing short of superb. But it does have some limitations: can be SLOW for continuous tone color, does not support Postscript (yet - they "tell me" that a 3rd. party is working on this), fonts can be a problem but not if you store them as image files when used as text on micrographs, line drawings can get excessive cases of the "jaggies" from the mechanical drive ( only shows up on images if you magnify } 5X or so. There is a very objective review of this printer in the August 1993 edition of "Advanced Imaging" Vol 8 No 8 pp 37 - 39. P.S. If you use a Mac you'll need to purchase a $199 interface.
Peter Ingram ingram-at-rcc.rti.org =============================== 3)
Some time ago we had a demo of the Fargo Primera (wax transfer) printer and compared it to the Tektronix Phaser IIID which we have on site. The system demoed only had a three colour system so the blacks were not great. A 3 colour + black system is available. The ouput was not quite as good as the Tek. but when you consider that the Fargo was around a quarter of the price (in Australia) it was pretty good value. I have seen the dye sub. output but not had a chance to do direct comparisions.
I think that Byte reviewd the printer not long ago (or it may have been another computer journal).
I hope this is of some use.
Colin V.
John Fournelle Electron Microprobe Lab Internet:johnf-at-geology.wisc.edu Dept of Geology & Geophysics Telephone: (608) 262-7964 University of Wisconsin Fax: (608) 262-0693 1215 West Dayton Street Amateur radio: WA3BTA/9 Madison, WI 53706 (14.030, 21.030 mHz)
Journal of Microscopy, Vol. 174, Pt. 1, April 1994, pp. 1 - 14.
STANDARD PERFORMANCE CRITERIA FOR ANALYTICAL ELECTRON MICROSCOPY
by S. M. ZEMYAN & D. B. WILLIAMS , Department of Materials Science & Engineering, Lehigh University, Whitaker Laboratory, . 5 E. Packer Ave., Bethlehem, PA 18015, U.S.A.
Summary Users of analytical electron microscopy lack easy-to-use standards for assessing the consistency and quality of analytical performance. We propose using a Cr thin film of known thickness to measure three important characteristics related to performance: the Cr K alpha peak-to-background (P/B) ratio, the X-ray spectrometer relative efficiency, and the spectrometer energy resolution. We used a Cr specimen to determine the instrumental factors which influence the P/B ratio, finding that the highest P/B ratios are achieved in scanning transmission mode at the highest available accelerating voltage. We present values of the P/B ratio, and the detector relative efficiency and energy resolution which can be used for comparison in other laboratories using the standard film.
Journal of Microscopy, Vol. 174, Pt. 1, April 1994, pp. 15 - 22.
ATOMIC FORCE MICROSCOPY IN THE PHOTOCHEMISTRY OF CHALCONES
by G. KAUPP , Universitaet Oldenburg, FB 9 - Organische Chemie I, Postfach 2503, D-26111 Oldenburg, Germany
Summary The application of atomic force microscopy (AFM) to photodimerization of crystalline chalcones provides new insights into the detailed mechanisms of solid-state reactions on the molecular level. Well-directed long-range transport phenomena are found which reach far beyond the crystal lattice distances. Reactions occur in the surface region where the light is absorbed. Characteristic features are built up that depend on crystal structure and crystal face. This could not be foreseen by previous theories based solely on a topochemical postulate/principle. There is now a much more intimate correlation of crystal structure with solid-state reactivity and this is directly studied and proven experimentally by AFM. Even solid-state reactions which are in opposition to topochemistry can be studied and understood on a molecular basis. The three-dimensional resolution of undisturbed insulating surfaces which is obtained by AFM is not available by any other technique.
Journal of Microscopy, Vol. 174, Pt. 1, April 1994, pp. 23 - 30.
REGRESSION METHODS FOR AUTOMATED COLOUR IMAGE CLASSIFICATION AND THRESHOLDING
by E. J. BREEN , CSIRO, Division of Mathematics and Statistics, Institute of Information Science and Engineering, Locked Bag 17, North Ryde NSW 2113 Australia
Summary Regression methods are used to perform automated image thresholding and colour pixel classification. This is done by considering threshold levels and pixel classification labels as pattern attributes. A regression equation that performs a mapping from the J dimensional feature-pattern space to the K dimensional attribute space is derived. The approach is non-parametric and deterministic, hence no assumptions about the statistical properties of the input patterns or images need be made. Initially a known set of input patterns with associated attributes are used to constitute a training set. A mapping function is then determined from the training patterns and used for estimating attribute values from unknown input patterns, such as images.
Journal of Microscopy, Vol. 174, Pt. 1, April 1994, pp. 31 - 38.
LIP-MODEL-BASED THREE-DIMENSIONAL RECONSTRUCTION AND VISUALIZATION OF HIV-INFECTED ENTIRE CELLS
by P. GREMILLET,* M. JOURLIN + & J.-C. PINOLI ++ , *Misis Image, BP 711, 42950 Saint-Etienne Cedex 9, France. + Laboratoire Lyonnais des Signaux et Systemes, Ecole Superieure de Chimie, Physique et Electronique, 31 Place Bellecour, 69288 Lyon Cedex 02, France, and Laboratoire de Traitement du Signal, Universite de Saint-Etienne, 23 Rue Paul Michelon, 42023 Saint-Etienne Cedex 02, France. ++Pechiney, Centre de Recherches, BP 27, 38340 Voreppe, France
Summary This paper presents a global solution from acquisition to visualization for the three-dimensional reconstruction of cell sections. Original techniques are proposed for the correct handling of the geometrical section distortions, and a new interpretation based on the logarithmic image processing (LIP) model is applied in order to create normalized grey-level sections where these are missing. Finally, a new method for generating a mesh of triangles to describe the envelope of the reconstructed cell is proposed, as well as a visualization mixing image synthesis and grey-level information. The product allows the user to explore the reconstructed cellular block in any desired direction, by showing user-defined grey-level sections inside the block mixed to a synthetic view of the cell envelope.
Journal of Microscopy, Vol. 174, Pt. 1, April 1994, pp. 39 - 46.
SURFACE ULTRASTRUCTURE OF OLFACTORY RECEPTOR SENSE HAIRS IN THE SILKMOTH, ANTHERAEA PERNYI
by V. I. POPOV, A. A. NIKONOV, N. K. AGAFONOVA & E. E. FESENKO , Institute of Cell Biophysics, Russian Accademy of Sciences, Pushchino, Moscow Region, 142292, Russia
Summary The fine structure of silkmoth sense hair surfaces has been investigated by freeze etching with Pt/C rotary shadowing. To do this, the hydrophobic layer in the cuticle was removed using 20 - 25% methanol or ethanol. Freeze-etch patterns of sense hair surfaces, as well as the pore structure and pore distribution, are shown. The hair surface has a nonhelical `band' structure, in which every `band' lies at an oblique angle with respect to the axis of the hair. Each `band' is separated from its neighbour by a 30-nm step. The average density of pores is 11.3 plus or minus 2.4 pores per square micrometre. Freeze-etch patterns of the single and multiple pore tubules are shown. Evidence for direct contact between the pore tubule and dendrite membrane of an olfactory receptor neuron is presented.
Journal of Microscopy, Vol. 174, Pt. 1, April 1994, pp. 47 - 50.
AGAR STANDARDS FOR QUANTITATIVE X-RAY MICROANALYSIS OF RESIN-EMBEDDED PLANT TISSUES
by E. FRITZ & G. JENTSCHKE , Institute of Forest Botany, University of Gottingen, Busgenweg 2, 37077 Gottingen, Germany
Summary Calibration standards for quantitative X-ray microanalysis of resin-embedded plant tissue were prepared by adding 6 - 600 mm KC1 to 5% agar. Agar blocks with an edge length of 1 - 2 mm were rapidly frozen, freeze-dried and embedded in styrene-methacrylate. Dry sections 1 micrometre thick were mounted on adhesive-coated grids. Apart from fine-scale inhomogeneities caused by ice crystal formation, the KC1 is evenly distributed in the agar blocks. The peak-to-continuum values of K and Cl were highly linearly correlated to the K and Cl contents over the whole concentration range.
Journal of Microscopy, Vol. 174, Pt. 1, April 1994, pp. 51 - 54.
A SIMPLE METHOD FOR PROCESSING INDIVIDUAL OOCYTES AND EMBRYOS FOR ELECTRON MICROSCOPY
by C. NOGUES, M. MARTI, M. BOADA & M. PONSA , Departament Biologia Cel.lular i Fisiologia, Facultat Ciencies, Universitat Autonoma de Barcelona, 08193-Bellaterra (Barcelona), Spain
Summary A simple method for handling individual specimens that must be processed either for scanning or transmission electron microscopy studies is described. For scanning microscope processing, dehydration is carried out with samples enclosed in small cages made from TAAB capsules in which top and bottom are substituted by plankton nets, and for transmission electron microscopy, samples are pre-embedded in agarose. This procedure significantly reduces mouth pipetting, dissecting microscope observations, is less labour intensive and, most importantly, reduces sample loss.
Journal of Microscopy, Vol. 174, Pt. 1, April 1994, pp. 55 - 58.
ENHANCED RETENTION OF MAGNETIC PARTICLES (E.G. MICROTOMED SECTIONS) IN A TEM
by W. A. FURDANOWICZ & K. E. DOWNEY , Homer Research Labs, Bethlehem Steel Corporation, Bethlehem, PA 18016-7699, U.S.A.
Summary An electron-transparent layer of adhesive is used to restrain magnetic particles from being stripped off the support film by the magnetic field of an objective lens in a TEM.
ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ RAPID PUBLICATION Journal of Microscopy, Vol. 174, Pt. 1, April 1994, pp. RP1 - RP 2.
MONOMERIC COLLAGEN IMAGED BY CRYOGENIC FORCE MICROSCOPY
by M. B. SHATTUCK, M. G. L. GUSTAFSSON, K. A. FISHER, K. C. YANAGIMOTO, A. VEIS, R. S. BHATNAGAR & J. CLARKE
IF YOU ARE INTERESTED IN SUBMITTING A MANUSCRIPT TO THE JOURNAL OF MICROSCOPY, OR HAVE ANY OTHER QUESTIONS ABOUT THE JOURNAL, PLEASE CONTACT DR GILLIAN WILSON, JOURNAL OF MICROSCOPY EDITORIAL OFFICE, 37/38 ST CLEMENTS, OXFORD, OX4 1AJ, UNITED KINGDOM. TELEPHONE +44 865 248768, FAX +44 865 791237, EMAIL RMS-at-VAX.OX.AC.UK.
AA Happy EA H ÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄÄ
A belated reply to the person asking about cat skin. In felis domesticus the skin has all of the same layers as found in human skin. However, the skin is thinner because there are only a few layers of dermins (2-4 layers) as compared to the {10 layers found in most areas of human skin.
We also have been considering CD-ROM for archival and image transfer. I have been told that only recently some of the authoring drives allow reading a partially filled platter, and then only on the authoring drive, not on a standard reading-only drive. I was hoping to use CD-ROM as an alternative to buying magneto-optical drives for portable data, as oppposed to setting up a server.
What has anyone experienced or read about this?
Glen MacDonald Hearing Development Laboratories RL-30 University of Washington Seattle, WA 98195 (206)543-8360 glenmac-at-u.washington.edu
On Fri, 25 Mar 1994, Russell E. Cook wrote:
} Michel Deschuyteneer suggested CD-ROM as a method for archiving images. } This is something we have been trying to do for the last few months. If } you plan to purchase a CD writer, you need to be aware that you may also } need to buy a high speed hard disk from which the images will be downloaded } to the CD. Look carefully at the input specifications of the CD writer } before you buy the high speed disk. } } } Russell E. Cook } Electron Microscopy Center for Materials Research } Materials Science Division } Argonne National Laboratory } Argonne, Illinois 60439 } USA } } }
If all people are talking about is archiving, then I would strongly suggest that DAT tapes are considered. The prices of the hardware for CD-ROM and DAT are comparable, but the price of a single tape is $16 and it can hold 2-16 Gbytes of data. The only reason in my mind for using CD-ROM is if you want additional disc space which is less expensive than true hard disks; for archiving it may well be a waste of money.
As Robin Wright says, CDs are forever. Also, they allow random access and it is extremely cheap to buy an internal CD-ROM player with new computers
Glen
On Mon, 28 Mar 1994, L. D. Marks wrote:
} If all people are talking about is archiving, then I } would strongly suggest that DAT tapes are considered. The } prices of the hardware for CD-ROM and DAT are comparable, } but the price of a single tape is $16 and it can hold } 2-16 Gbytes of data. The only reason in my mind for using } CD-ROM is if you want additional disc space which is less } expensive than true hard disks; for archiving it may well } be a waste of money. } } Laurie Marks, NU }
Message-Id: {MAILQUEUE-101.940329082400.480-at-FS-IAM-1.JRC.NL} To: microscopy-at-anlemc.msd.anl.gov
I agree that DAT tape is not ideal for archiving due to eventual degradation etc, but what about these new Phase-Change optical drives? I don't know much about them, except that they are re-writeable, cost about half that of CD-ROM writers (at least in Europe) and can store approx. 1Gb. I think that they are sold by Panasonic over here. Maybe somebody else knows more about them. As far as I can see they appear to have significant advantages, and are based on Sony's mini-disk technology, so the prices should drop fairly soon if that is a success.
Doug Arrell Mechanical Performance and Joining Institute for Advanced Materials 1755 ZG Petten Netherlands
Subject: Time:8:16 AM OFFICE MEMO CD archiving Date:3/29/94 Just my two cents worth on archiving on optical discs, CDs or DAT. We have been looking into CD archiving and feel that it has several advantages over the other two formats. 1) CDs allow relatively fast and easy acess to the images/files. So if you are planning to manipulate these images (of course you will have to work on copys placed on your hard disk) or require frequent acess to them this method is about equal to opticals, but superior to DAT, 2) Both CDs & opticals will have a longer expected life then DAT before media breakdown, and 3) the most important consideration for us is that CD readers are cheap, going down in price and are incorporated into the hardware of an increasing number of computer systems. This is important since every potential user is already equipped or makes only a small investment ( {$300 for many readers). The larger number of potential users will make the system more cost effective and may partially offset the initially higher cost of a CD writer. A further consideration is that both Mac and IBM clone users need acess. The formatting for opticals is usually specific to one or the other of these standards (unless you plan to go with Apple PowerPCs) and will thus be more limited. Photo CDs can be read on drivers of either machine format with no conversions or translations required. We think that for these reasons, CDs are the way to go.
I have been requested to pass on the following message for discussion:
From: Bill Monroe Electron Microscope Center Mississippi State University
His Question: Occasionally our laboratory staff has experienced problems with thin sections coming off the grid during immuno-gold labeling steps. The tissue sections are from L.R. White embedded material and are mounted on 200 mesh formvar-coated nickel grids.
The Question: Are there any suggestions to remedy this problem of sections coming off the grid?
Richard Kuklinski, rfk2-at-ra.msstate.edu Electron Microscope Center, Mississippi State University
How do I persuade Image to measure the number of intercepts on a line or to measure the length of individual intercepts? Both of these measurements are useful in determining the grain size of metals, for example, see ASTM standard E112 and E1181.
We originally posted this to the NIH-Image list but received no responses, so if anyone can help, we'd appreciate it. Thanks.
You can reply to my e-mail address smiths-at-mlc.lib.mi.us OR directly to my patron at the following address:
Sam Purdy National Steel Corp. Technical Research Center 1745 Fritz Drive Trenton, MI 48183 313/676-2682 or FAX:313/676-2030
I am looking for the name and address of the manufacture of crystalbond. We had purchased in the past sticks about 3/4" in diameter at a very reasonable price (much below Gatan's price) but can't locate the source now.
Serial Sections? An easy way to keep serial sections attached to one another is to coat the block with a sticky substance. Previously, I used something called "Takki-wax". Does anyone know of a supplier for this wonderful stuff, we just ran out?
We are interested in collecting RHEED and LEED images with a video camera. Stuff like line profiles, spot intensity fluctuations. I checked into this several years ago and found a system by Staib Instruments It was a lot of money. I then thought of buying a system with a Sony XC-77 camera, Fuji CF50L lens, DT2851 Framegrabber, Halo 88, and Image Pro software. This sytem would have cost about $7K. Several years have past and now we have a Mac Quadra 840AV. Someone here thinks he can do the same thing with the Mac AV inputs that these other systems could do with dedicated boards. Any comments from people running such systems would be helpful.
} } Serial Sections? } An easy way to keep serial sections attached to one another is to coat the } block with a sticky substance. Previously, I used something called } "Takki-wax". Does anyone know of a supplier for this wonderful stuff, we just } ran out? } ================================================================
I have also heard of using diluted rubber cement for keeping serial sections together. ********************************************************** * Greg Erdos ** * * Director, ICBR EMCL ** Phone 904-392-1295 * * 218 Carr Hall ** FAX 904-392-8598 * * University of Florida ** gwerdos-at-gnv.ifas.ufl.edu * * Gainesville, FL 32611 ** * **********************************************************
We've been using the Becton-Dickinson mouse monoclonal antibody againsty bromodeoxyuridine (BrdU) for a few years for a variety of samples and fixatives. We'vew been using at 1/4000 successfully for ABC-peroxidase and fluorescently labelled secondary antibodies.
The BD is very pricey, $245 per bottle. Has anyone tried other brands of antibodies or fluorescently conjugated anti-BrdU antibodies? What dilutions did you use, how sensitive did you find them (especially the conjugated Ab)?
Glen MacDonald Hearing Development Laboratories RL-30 University of Washington Seattle, WA 98195 (206)543-8360 glenmac-at-u.washington.edu
Message-Id: {9403291926.AA22920-at-riker.ml.wpafb.af.mil} To: WALCKSD:ml:wpafb, SCHELTFJ:ml:wpafb Subj: Crystalbond Orig-Author: {"M. J. Kramer" {MJKRAMER-at-vaxld.ameslab.gov} }:ddn:wpafb ----------------------------------------------------------- I am looking for the name and address of the manufacture of crystalbond. We had purchased in the past sticks about 3/4" in diameter at a very reasonable price (much below Gatan's price) but can't locate the source now.
Re} I-G-labeled sections falling off Are the sections falling off or is it the formvar that is comming away from the grids? I've never had any problem with sections comming detached from formvar-carbon films.
} } I have been requested to pass on the following message for discussion: } } } From: Bill Monroe } Electron Microscope Center } Mississippi State University } } His Question: Occasionally our laboratory staff has experienced } problems with thin sections coming off the grid during immuno-gold } labeling steps. The tissue sections are from L.R. White embedded } material and are mounted on 200 mesh formvar-coated nickel grids. } } The Question: Are there any suggestions to remedy this problem of } sections coming off the grid? } } } Richard Kuklinski, rfk2-at-ra.msstate.edu } Electron Microscope Center, Mississippi State University ========================================================== REPLY
We have never experienced this problem. We do however pick up sections from above rather than from below thereby eliminating the possibility of trapping water between the section and the formvar film. I don't know if this would make any difference. ********************************************************** * Greg Erdos ** * * Director, ICBR EMCL ** Phone 904-392-1295 * * 218 Carr Hall ** FAX 904-392-8598 * * University of Florida ** gwerdos-at-gnv.ifas.ufl.edu * * Gainesville, FL 32611 ** * **********************************************************
Crystal bond is supplied and made by Universal Photonics 1-800-645-7173. Universal Photonics is a supplier of materials for lens grinding and polishing (used to be Universal Optics) and developed Crystal bond for holding prisms and crystals to the work tool as they are being lapped. Their catalog has a lot of interesting stuff of potential use to microscopists, but for years was out of print. I think you can get one now, however.
South Bay Technology produces 4 standard mounting waxes designed for mounting samples for precision lapping, polishing and cutting. SBT makes a wax which is essentially identical to Crystalbond 509. The only difference is that QuickStick 135 is very clear and does not have the amber color normally associated with Crystalbond.
QuickStick 135 is acetone soluble and has a flow point of 135 degrees C. QuickStick 135 is used extensively in TEM sample preparation operations and is supplied with all South Bay Technology TEM sample preparation equipment. Quick Stick 135 is packaged in a plastic tray consisting of 20 3" x .25" x .25" unwrapped sticks (Approx. 360 grams) for $40. It can also be supplied in the 7" long cardboard tubes as is Crystalbond or in a variety of other forms.
For information on our other mounting waxes, free wax samples or information on any of our sample preparation products, please contact me as follows:
David Henriks South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
TEL: 714-492-2600 Toll-free in USA: 800-728-2233 FAX: 714-492-1499
e-mail: via CompuServe at: David Henriks, 73531,1344
Bill Monroe asks about sections falling off of grids during immunostaining. I use uncoated gold grids (no grid induced astigmatism) with LR White or Spurr's resin. I section blocks, pick up sections and let dry at least one day before immunostaining. Once in a while a few sections might fall off but this is not a major problem. I also immerse the grids in all solutions on parafilm. Hope this helps.
Anybody out there have suggestions for measurement of particles from the TEM? The particles are thought to measure about 80 angstroms. We are assuming using 80-200,000x magnification and need very precise measurements. The commercial standards are only good up to 50,000x.
I appriciate any help!
Kathy Walters Central Electron Microscopy Researh Facility University of Iowa
Kathy, How about using carbon graphite at 3.4 Angstrom lattice spacing or there is a type of asbestos that gives 9 Angstrom lattice spacing. These could be used at 100KX or above and the mag accuracy of your TEM could then be calculated. Cal
Message-Id: {9404011318.AA00750-at-odin.morph.med.umich.edu} Errors-To: {dennis%odin-at-odin.morph.med.umich.edu} To: microscopy-at-anlemc.msd.anl.gov Cc: dennis-at-odin.morph.med.umich.edu
On Fri, 1 Apr 1994 dennis%odin-at-odin.morph.med.umich.edu wrote: Snip } My lab is looking for a citation in the literature about the stability of } specimens over time in polymerized Epn blocks. I've checked the literature } but have not had much luck. If anyone knows offhand where this can be find, I } would appreciate the reference.
Hi, I've recently been reading a book called _Artifacts iin Biological Electron Microscopy_ by R.F.E. Crang and K.L. Klomparens, Plenum Press, 1988, and have found much of the info to be very applicable to some of my general problems. There is a section by H.H. Mollenhauer that discusses dehydration and epoxy embedding that might have the info you're looking for. Good luck.
Dwight U. Beebe beebed-at-ere.umontreal.ca Institut de recherche en biologie vegetale Voice:514-872-4563 4101, rue Sherbrooke est FAX:514-872-9406 Montreal, PQ H1X 2B2 Canada
Picking up on Cal's idea, would it be possible for you to prepare your particles right on top of the carbon graphite, on Formvar coated grids for support. That way you could photograph them together in the same micrograph and get the highest accuracy measurement of the particle sizes by direct comparison to the carbon lattice image.
The question I have, is can one compare the particles directly to the image of the crystal lattice spacings, or is there a calculation that must be done to correctly use that spacing information? Anyone? Gib Ahlstrtand.
------------ Forwarded Message begins here ------------
Kathy, How about using carbon graphite at 3.4 Angstrom lattice spacing or there is a type of asbestos that gives 9 Angstrom lattice spacing. These could be used at 100KX or above and the mag accuracy of your TEM could then be calculated. Cal ---------- Forwarded Message ends here ------------
-- Gib Ahlstrand 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
In reply to Kathy Walters query about resolution measurements, try using a colloidal gold preperation, say a 5nm gold. The makers of the probes send along info which contains a distribution curve of the size of the gold. Hope this helps.
After finding a suitable calibration standard, you could use one of the video measurement systems on the market to do the actual measurements. The standard is used to calibrate the measurement system only, and does not have to be closely related to the size of your sample. This would still give very accurate results, down to a fraction of the size of the standard.
My company makes such a device, called the XR-2000. It has a very large array of measurements, like distance, area, angle, pathlength, etc, and lots of other functions. It is very inexpensive and easy to use.
Your system must use standard video (like Y/C or RGB) for this to work.
If you would like to know more about the XR-2000, private Email to me.
Message-Id: {9404041513.AA02301-at-odin.morph.med.umich.edu} Errors-To: {dennis%odin-at-odin.morph.med.umich.edu} To: microscopy-at-anlemc.msd.anl.gov Cc: dennis-at-odin.morph.med.umich.edu
Greetings! I will teach a comprehensive, semester-long TEM course next Fall and am trying to find a good text. My choice at the moment is: Electron Microscopy. Principles and Techniques for Biologists by J.J. Bozzola and L.D. Russell. Does anyone have other favorite texts? The course will be small (6 graduate students) and will include both theory and practical sessions. Both plants (my specialty) and animals will be used as material. I would also be very interested in hearing directly from people who have taught such a course or one similar to it, for tips and "what works, what doesn't" info. We will also use the RMS Handbook: The Operation of Transmission and Scanning Electron Microscopes by D. Chescoe and P.J. Goodhew, as we have a JEOL 100S for the course and this book has a nice description of alignment procedures which match our instrument. Thanks in advance, any help will be greatly appreciated.
Dwight U. Beebe beebed-at-ere.umontreal.ca Institut de recherche en biologie vegetale Voice:514-872-4563 4101, rue Sherbrooke est FAX:514-872-9406 Montreal, PQ H1X 2B2 Canada
} In reply to Kathy Walters query about resolution measurements, try } using } a colloidal gold preperation, say a 5nm gold. The makers of the } probes } send along info which contains a distribution curve of the size of } the gold. Hope this helps.
Colloidal gold is easy to see and measure, but is not all that uniform. As Ed says, the manufacturers send along a size distribution curve. The sizes - diameters and distribution - will vary from batch to batch. One would also want to know how the manufacturer's size distribution was determined. They presumably would also have to measure the sizes microscopically, and the data would not need to be really accurate to suit their purposes.
The Journal of Microscopy has another few new books for review. Interested parties should contact Gillian Wilson at RMS-at-VAX.OX.AC.UK.
The books are:
1. Introduction to Scanning Tunnelling Microscopy. By C Julian Chen. OUP, 1994. 412 pages.
2. Polymerase Chain Reaction. By C R Newton & A Graham. BIOS in association with the Biochemical Society, 1994. 161 pages.
3.Rapid Methods and Automation in Microbiology and Immunology. Edited by R C Spencer, E P Wright & S W B Newsom. Intercept Ltd, France, 1994. 502 pages.
Subject: Time:2:05 PM OFFICE MEMO Re:EM Text Date:4/5/94 Another good text to consider might be "The Principles & Practice of Electron Microscopy" by Ian M. Watt, Cambridge Univ. Press. It is very well written and covers a good range of subjects for a one-term course.
Subject: Time:2:09 PM OFFICE MEMO Re: SEM Screen color Date:4/5/94 I think one justification for using green phosphors is that the human eye has maximum sensitivity in the green range of the visible spectrum.
Additional note: to avoid to thick a formvar which can remain between grid bars, use a quick blast of compressed air directed the grid to vaporize formvar between the grid bars.
On Fri, 1 Apr 1994 wrightr-at-zoology.washington.edu wrote:
} My very first immunolabeling experiment ended when every section } floated off the 25 grids that I was labeling at the time. Since } then, I use a method shown to me by Bonnie Chojnacki. Specifically I } use grids that have been dipped in a dilute formvar solution (about } 0.1% in chloroform) and then dried on filter paper. The grids can } either be dipped individually into the formvar and then placed on } filter paper, or a whole canister can be dumped into the formvar and } each grid retrieved individually onto filter paper. } } } This procedure produces a sticky coat on the grid bars and sections } (epoxy, LR-white, etc) really stay attached throughout all of the } incubations. } } } } } Robin Wright }
The answerto Your questionK about the same information after 30 year depends on what information you want. I wouldnt trust immunocytochemistry of material embedded that long, since we see a difference even after a month (post embed stain of etched epon). However, the structures have not changed and quantitation done will be the same. Do you need a reference if the experience of a great many microscopists says there is no difference?
On Mon, 4 Apr 1994 dennis%odin-at-odin.morph.med.umich.edu wrote:
} Thanks to everyone who responded the my question. However, most of the } citations sent had to do with the stability of sections under beam current, } heat, etc... } } The situation is that a drug company that has provided a grant to us would } like to know for certain that the tissue that has been embedded in epon can be } pulled out from storage thirty years and will still be good. Will the tissue } be sectionable and will it still give the same data as before? } } As I said, most of the people here still have blocks that they did their } undergraduate work in back in the 1960s, and they are still as sectionable as } the day they were pulled out of the oven. In fact, I understand that the } quality of epon actually improves over time because it will have longer to } fully polymerize. } } So I am then simply looking for a citation on something that we already seem } to know, but still have not been able to find it. } } Thanks again, } Dennis Shubitowski } dennis-at-odin.morph.med.umich.edu }
Green phosphor has a greater luminous efficiency, and so can be made much brighter than white phosphor with equivalent electron optics. This makes a green TEM screen useful, but does not make a good argument for a green monitor in a darkened lab. The SE M manufacturer probably chooses a phosphor based on decay time. The green P20 phosphor has a decay time similar to that of the human eye, and so "integrates" the noisy signal optimally.
regards Mark W. Lund, Director MOXTEK, Inc. Orem UT
} I'm looking for a good inexpensive seconhand ultramicrotome similar } or identical to RMC MT-7000 ULTRA or REICHERT ULTRACUT S.
I don't have an Ultracut S., but I do have a Reichert Jung Histocut II that has never been used, and we are now considering selling it. It was purchased for a project that never materialized, and it has never actually cut anything.
As Nestor indicated several times before, and also in the introductory welcome material sent to new subscribers, is important including in your message the suject header. It allows determining nature of message without having to read the content. Given the increased usage of the service, inclusion of the subject header is a nice gesture toward other users. In fact, many more will probably not unsubscribe.
Cesar D. Fermin, Ph.D Tulane Medical School Pathology/SL79 New Orleans, La 70112
Hi, I have not used Image, but have tried to use another image analyser to measure the number of intercepts on a line and the intercept lengths in the past. The general procedure was as follows: 1) Use grey level to differentiate your grains from the grain boundaries and create a bit map of the grains. 2) Decrease your active measurement field until you have only a single rastor line across the screen. 3) If only the "detected" portion of your image is overlayed with a colored "bitmap", then you should have a colored broken line across the screen at this point. 4) Count the objects detected to give you a measure the number of intercepts. 5) Take the maximum dimension of the objects detected to give you a measure of the intercept length. 6) By rotating the image and repeating the procedure, information may be collected in relation to variation with orientation.
Hope this is of assistance. ______________________________________________________________________ Laurie Frederick, A.SC.T. PAPRICAN Corrosion Control Group 3800 Wesbrook Mall The Pulp and Paper Research Vancouver, B.C. Institute of Canada Canada V6S 2L9
I am trying to locate an e-mail address for Dr. Mark E. Welland -at- Cambridge University, Engineering Dept., St. John's College. Last I knew, he was working in Scanning, Tunneling Microscopy. If anyone knows of his address (or, if you're on this list, Mark) please respond directly to my e-mail address. Thanks very much!
Susan K. Smith National Steel Corp. smiths-at-mlc.lib.mi.us
Hi, Sorry to broadcast this, but I deleted the message before I realized I might have an answer. A request was made about a source for a OM-3 motor. I checked with a company I buy equiment from and was told they have several on hand. The company is Marivac, Ltd., 5821 Russell St., Halifax, Nova Scotia, B3K 1X5 Canada. 800-565-5821, FAX 902-455-4007. I have purchased equipment from these folks (Balzers MED 010) and have a service contract with them for my Ultracut. The person I deal with is Chris Cathcart; he operates out of Toronto (416-495-0389), but can be reached via the 800 number. I've been pleased with the quality of the service I get. I have no connection with this company other than as a satisfied customer, standard disclaimers apply, blah, blah, blah. Good luck!
Dwight U. Beebe beebed-at-ere.umontreal.ca Institut de recherche en biologie vegetale Voice:514-872-4563 4101, rue Sherbrooke est FAX:514-872-9406 Montreal, PQ H1X 2B2 Canada
Anybody have a good fixative for cross-linking polysaccahridases? I'm trying to do SEM on a "gliding" bacteria strain and find my bacteria being lifted off the agar. I've tried vapor fixation using osmium and then fixing overnight in 2% glut. One colony strain will lift off the agar surface and the other will remain. Any Suggestions?
I have used 50 X 9 mm culture dishes with friction fit lids manufactured by Simport Plastics, Beloeil, Quebec, Canada (514) 464- 1723. The catalogue number was D210-14.
I would be interested in a source of smaller diameter dishes with a larger internal clearance for storing my metallographic samples.
Hope this is of assistance. ______________________________________________________________________ Laurie Frederick, A.SC.T. PAPRICAN Corrosion Control Group 3800 Wesbrook Mall The Pulp and Paper Research Vancouver, B.C. Institute of Canada Canada V6S 2L9
Falcon 35x10mm dishes (available through Fischer) work for embedding cultured cells.
Another possibilty is to grow the cells on culture inserts (available from Millipore, Falcon, and other companies) which are placed into 24 well plates. After fixation (or post-fixation if you prefer), the filter on the bottom of the insert can be removed with a razor blade, and the filters dehydrated with ethanol and embedded. This uses even less reagents that the 35x10mm dishes, and the 24 well plate is easier to manipulate than several round dishes. I've used this technique for culturing fetal kidneys and for endothelial cells.
Hello, Our laboratory is in the process of investigating ion mills. It appears that Baltec, Fischione and Gatan are the principle manufacturers of this piece of equipment. Any comments, suggestions, experiences or thoughts about ion mills would be appreciated. Thanks, John
In answer to Phil Rutledge's query about fixation of "polysaccahridases": The closest thing I know to a fixative for polysaccharides is Alcian Blue. This works reasonably well in most cases where bacterial capsules need to be insolubilised. Try the following refs: Tuffery - 1969, J gen Microbiol 57:41-50 Goldberg & Septier - 1986, Anat Record 216:181-190 Scott - 1972, Histochemie 32:191- (Primary reference) Trachtenberg - 1986, J Ultrastruct Mol Res 97:80-102 Steedman - 1950, Quart. J Microsc Sci 91:477-479 Powell et al - 1992, J Fish Biol 41:813-824 (useful ref) Dellorbo et al - 1992, J Electr Microsc 41:475-479.
Hope this is of some use.
Jan Coetzee Unit for Electron Microscopy Tel:+27-12-420-2075 University of Pretoria Fax:+27-12-342-1738 Pretoria Internet:janc-at-ccnet.up.ac.za 0002 South Africa
Message-Id: {9404111414.AA06580-at-riker.ml.wpafb.af.mil} To: WALCKSD:ml:wpafb, SCHELTFJ:ml:wpafb Subj: ion mills Orig-Author: {PHELPS-at-ENH.NIST.GOV}:ddn:wpafb ----------------------------------------------------------- Hello, Our laboratory is in the process of investigating ion mills. It appears that Baltec, Fischione and Gatan are the principle manufacturers of this piece of equipment. Any comments, suggestions, experiences or thoughts about ion mills would be appreciated. Thanks, John
Phil Rutledge writes: } Anybody have a good fixative for cross-linking polysaccahridases? I'm } trying to do SEM on a "gliding" bacteria strain and find my bacteria being } lifted off the agar. I've tried vapor fixation using osmium and then } fixing overnight in 2% glut. One colony strain will lift off the agar } surface and the other will remain. Any Suggestions? } Phil,
Here is a well worn ruthenium red staining method for mucopolysaccharides from M.A. Hayat: "Positive Staining for Electron Microscopy", p. 169-170. Whether this will fix polsaccahridASES, or prevent them from sliding off the agar, I cannot say.
A few preliminary notes from Hayat: 1. Maximum contrast obtained when ruthenium red and osmium tetroxide applied together(refering to TEM prep here). 2. Stain can be applied by adding it to fixative solution, water, or buffer solution. 3. Phosphate buffers are not recommended for ruthenium red-osmium tetroxide procedures. Chloride-free cacodylate buffer considered most ideal one to use.
For general purposes, the following method of fixation and staining for acid mucopolysaccharides recommended:
Solutions: A. Aqueous glutaraldehyde (4%) 5 ml 0.2 M Cacodylate buffer (pH 7.3) 5 ml Ruthenium red stock solution 5 ml (1,500 ppm in water)
B. Aqueous osmium tetroxide (5%) 5 ml 0.2 M Cacodylate buffer (pH 7.3) 5 ml Ruthenium red stock solution 5 ml (1,500 ppm in water)
Mix solution B just prior to use. 1,500 ppm is 30 mg ruthenium red in 20 ml water. Fix and stain samples in solution A for 1 hr. at room temperature. Rinse briefly with buffer, then post-fix and stain with solution B for 3 hr at room temperature. Rinse briefly with buffer, then dehydrate and embedd, or critical point dry(in your case) according to standard procedures.
Ruthenium red stain is available from most EM or chemical suppliers.
The above is sort of the mother of all ruthenium staining techniques for EM. Others have since modified its use and here are more recent references:
1. B. Giammara and J. Hanker, Ruthenium Red-osmium bridging with TCH: New techniques to stain biological specimens for light and TEM and to coat them for SEM, Proceedings of the 46th Annual Meeting of the Electron Microscopy Society of America, 1988, pp20-21.
2. M. Jacques and L. Graham, Improved preservation of bacterial capsule for electron microscopy, Journal of Electron Microscopoy Technique 11:167-169 (1989). This paper compares ruthenium-glutaraldehyde staining to glutaraldehyde-amine, which they found to be superior.
The following papers are on non-ruthenium red methods, and involve TCH (thiocarbohydrazide) and tannic acid procedures:
3. R.O. Kelley, R. Dekker and J. Bluemink, Ligand-mediated osmium binding: Its application in coating biological specimens for SEM, J. Ultrastructural Research, 45:254-258 (1973). (The classic reference in applying TCH for SEM)
4. J. Murphy, Non-coating techniques to render biological specimens conductive/1980 update, Scanning Electron Microscopy/1980/I, pp209-220.
-- Gib Ahlstrand 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
This Master Class is intended for novices, experienced specimen preparers and scientists involved in the examination of transmission electron microscopy specimens. The course will present the latest materials, tools, and methods for preparing high-quality specimens.
Description
While most aspects of specimen preparation will be covered, emphasis will be placed on preparing cross section specimens of complex composite samples, such as those found in the semiconductor industry; mechanical polishing to transparency; and ion milling. The course will review a variety of modern physical science specimen preparation methods, discussing the advantages, disadvantages, and limitations of each. Experience has shown that a wide range of bulk specimen configurations may be successfully prepared. Discussion will include the preparation of metals, ceramics, thin films and coatings, powders, wires, and difficult (small volume, radioactive, etc.) specimens. Laboratory demonstrations are planned.
Instructors
Ron Anderson; Senior Physicist at the IBM East Fishkill Facility. Vince Carlino; President of VCR Group. George Lane; Materials Product Manager for Bal-Tec, Inc. Jo Ellen Tyson; Applications Specialist for Mager Scientific.
Registration and Attendance
To register, call or e mail Kim Knudtson at 612 626 7594 (kimberly-at-maroon.tc.umn.edu). Enrollment is limited; please respond early to guarantee a space. Disability accommodations will be provided upon request. This publication and material is available in alternative formats upon request.
Tuition
The fee will cover course materials, refreshments, and box lunches. For members of the University community, the charge is $70; for all others, the fee will be $250. University of Minnesota attendees should provide a CUFS number, others should provide information so that they may be billed after the course.
Program Sponsor
The Center for Interfacial Engineering (CIE) is an NSF Engineering Research Center supported by the NSF, the University of Minnesota, and the corporate members of CIE. The CIE Characterization Facility and High-Resolution Microscopy Center provide instrumentation and personnel to facilitate the physical and chemical study of interfaces. These facilities are available to all members of the University community as well as external users. Tours of the labs will be available to interested attendees.
Accommodations
The University of Minnesota is located 10 miles from the Minneapolis-St. Paul International Airport. The Radisson Metrodome Hotel is conveniently located within two blocks of the lecture and laboratory sessions. The telephone number is 612-379-8888. A Days Inn is a 20-minute walk from the University; the telephone number there is 612 623 3999.
Schedule
Day 1, April 28
Registration (8:00 am), 210 Amundson Laboratories held in the High-Resolution Microscopy Center, Shepherd Laboratories
Lectures: Ron Anderson „ Strategic plan Initial processing from bulk samples Sawing and grinding, dimpling „ Final processing the thin specimen Ion milling/FIB methods Mechanical microthinning
Vince Carlino „ Specimen Prep by Dimpling and Ion Milling
George Lane „ High Performance Ion Beam Thinning of Solid State Materials
Day 2, April 29
Lectures: Ron Anderson „ Other Methods: Microtomy, Cleavage, Replication „ Cross-sectional specimen preparation The IBM East Fishkill Method
Demonstrations
„ Ron Anderson: Tripod polisher
„ Vince Carlino: VCR Dimpler Ion Beam Sputterer Model IBS/TM200S Ion Milling System
„ George Lane: RES 010 Rapid Etching System MED 020 Modular High Vacuum Coating System
„ Jo Ellen Tyson: Reichert Ultracut S/FCS cryo-ultramicrotome Attendees may bring their own samples.
Stuart McKernan stuartm-at-maroon.tc.umn.edu Chemical Engineering and Materials Science OR High Resolution Microscopy Center University of Minnesota, Minneapolis, MN 55455
we charge based on what it costs us: embed.....$6 thick sections (glass slide)....$16 Thin sections........$60 TEM time.............$36/hr TEM negative..........0.65 8X10 print............4.10 Publication quality print.....$5.50 Technician time if needed (microscope operation).......$25/hr
equipment costs included depreciation and service contract
hope this is helpful
On Mon, 11 Apr 1994, Tamara Howard wrote:
} A while back someone asked about the standard charges for a routine TEM } workup on a biological sample...I don't recall ever seeing any answers to that, } and now I am looking for similar information. What is considered to be a } reasonable charge for say one sample, plain old fix, embed, a few grids, and } maybe 2 hours max scope time plus prints? At an academic level, that is - I kno0w } clinical charges can be astronomical. } You can answer by e-mail to tah-at-med.pitt.edu if you do not want to } go through the usegroup on this. } I would really appreciate some help on this one! } Tamara Howard } U. Pittsburgh School of Medicine } Pittsburgh, PA }
I haven't had the chance to actually do it, but according to info. I received at the Bio-Rad Confocal course last month, it should be possible. The relative brightness of Cy3 may mean titrating its molar ratio relative to Cy5 and/or careful selection of filter blocks and PMT filters to prevent Cy3's long trailing slope from comming through the Cy5 signal. Take a look at Brelje et al, Chapter 4, Methods in Cell Biology, vol. 38, edited by Brian Matsumoto.
On Mon, 11 Apr 1994, Michael Cammer wrote:
} If you have a Kr/Ar laser, you can triple label with FITC, Cy3, and Cy5. } } }
} ratio relative to Cy5 and/or careful selection of filter blocks and PMT } filters to prevent Cy3's long trailing slope from comming through the Cy5 } On Mon, 11 Apr 1994, Michael Cammer wrote: } } If you have a Kr/Ar laser, you can triple label with FITC, Cy3, and Cy5.
We have not had a problem with Cy3 being imaged in the Cy5 channel (we image Cy5 only while exciting with the red laser line). However, we have had problems when imaging bright propidium iodide or Texas red with the standard long pass rhodamine filter block; some Cy5 appears to be excited.
We have generous institutional support and I am told that our charges are quite low. We do not have to recover any salary and about half our operational cost are from institutional support.
THESE ARE OUR IN HOUSE RATES. For outside clients I, at least triple the rates
The following charges are in effect at the EM Core .
Standard TEM or SEM sample $65.
Additional similar samples sub- mitted at the same time $55.
Immunolabeling or other cyto- chemical reactions per sample $75.
Negative stain per sample $35.
Partial preparation is also available with price appropriate to our input for example:
Fix and embed only $20/sample
Thin section only $20/block
Use of the microscopes by qualified users is $20 per hour and $1 per photograph. If you want Polaroid you must also supply the film.
For the benefit of the curious Materials Scientist like me, would someone please post a description (short) of what CY3/CY5 is and what it is used for.
I have been having problems with Quetol 651 resin for the past year, after using it successfully in my lab for 2 years. Has anyone else experienced difficulties? Of what sort? I'd like to correspond with someone with a background/knowledge of resin che mistry to possibly diagnose the defect.
Thanks to everyone who gave suggestions to the problem I am having with the fixation of the bacteria and having the cells float off of the agar. Now I'll have to sort through the suggestions and see what works best. Again, thanx to all, Phil Rutledge, Director Center for EM UMBC
Greetings, We just received 176 grams (ampules) of osmium tetroxide. The ampules, we are told, were bought some time back in the 1960's from Fisher, Englehard (in individual wooden cases!), National Lead, and MCB. The integrity of the ampules seems good as evidenced by the translucent greenish color of the osmium. However, the osmium seems to have coalesced into a single mass. Does anyone have any comments, warnings or advice? Considering that this gift is worth over $6K, I'd like to be able to use it.
************************************************* * * * * * Charles J. Butterick * * Electron Microscopy Center * * Department of Cell Biology and Anatomy * * Texas Tech University Health Sciences Center * * Lubbock, Texas 79430 * * USA * * Phone 806 743-2706 voice * * 806 743-2707 fax * * Email hecub-at-ttacs.ttu.edu * * * * * *************************************************
SOMEONE WROTE: In message {199404121522.IAA18262-at-ucsd.edu} writes: } I have been having problems with Quetol 651 resin for the past year, after } using it successfully in my lab for 2 years. Has anyone else experienced } difficulties? Of what sort? I'd like to correspond with someone with a } background/knowledge of resin che } mistry to possibly diagnose the defect. } } Hello,
We've used Quetol for years without trouble. You do not mention what problems you are having so I have no idea what to tell you. Put out another message on the MICROSCOPOY net describing what used to work well for you and what kind of trouble you are having now. That way I think people with experience will be able to zero in on what might work for you, and I will reply then. Thanks.
-- Gib Ahlstrand 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
Greetings, We just received 176 grams (ampules) of osmium tetroxide. The ampules, we are told, were bought some time back in the 1960's from Fisher, Englehard (in individual wooden cases!), National Lead, and MCB. The integrity of the ampules seems good as evidenced by the translucent greenish color of the osmium. However, the osmium seems to have coalesced into a single mass in each of the ampules. Does anyone have any comments, warnings or advice? Considering that this gift is worth over $6K, I'd like to be able to use it.
************************************************* * * * * * Charles J. Butterick * * Electron Microscopy Center * * Department of Cell Biology and Anatomy * * Texas Tech University Health Sciences Center * * Lubbock, Texas 79430 * * USA * * Phone 806 743-2706 voice * * 806 743-2707 fax * * Email hecub-at-ttacs.ttu.edu * * * * * *************************************************
Message-ID: {MAILQUEUE-101.940412174341.320-at-bmg.bhs.uab.edu} To: Microscopy-at-anlemc.msd.anl.gov
Does anyone know if there is a User's Group for the Macintosh-based image analysis software IP-LabSpectrum from Signal Analytics Corp.? I would be interested in linking up with users of that particular software platform to exchange ideas, information, scripts ect.
Thanks Kevin McCarthy Assistant Professor Department of Cell Biology University of Alabama at Birmingham Birmingham, Alabama 35294 Phone 205-934-9923/9924 Fax 205-934-7029
I notice from the literature that people setting up microscope-based ion measurement systems to quantitfy Na, Ca or H by fluorescence ratio microfluorimetry usually use a Xenon lamp for excitation of the fluoprophores. We are putting together a system for measuring Ca using Indo on an Olympus IMT-2 but lack a fluorescence source. The mercury lamp is half the price of the xenon, and certainly excites at the wavelengths we require (Indo exc. at 365 =B1 10nm), so we are thinking of going that way rather than the very expensive Xe.
Is the main advatage of Xe then the more constant output over the whole UV range so that dual excitation is more reliable (both excitation intensities the same, whereas with mercury one may fall over a peak and the other not) ? I'd appreciate comments from anyone who has experience in this area. Thanks, Ian Harper iharper-at-eagle.mrc.ac.za
I notice from the literature that people setting up microscope-based ion measurement systems to quantitfy Na, Ca or H by fluorescence ratio microfluorimetry usually use a Xenon lamp for excitation of the fluoprophores. We are putting together a system for measuring Ca using Indo on an Olympus IMT-2, but still lack a fluorescence source. The mercury lamp (100W) is half the price of the xenon, and certainly excites at the wavelengths we require (Indo exc. at 365+/- 10nm), so we are thinking of going that way rather than the very expensive Xe lamp.
Is the main advatage of Xe then the more constant output over the whole UV range so that dual excitation is more reliable (both excitation intensities the same, whereas with mercury one may exc wavelength may fall over/near one of the mercury peaks, whereas the other may not) ? I'd appreciate comments from anyone who has experience in this area. Thanks, Ian Harper iharper-at-eagle.mrc.ac.za
Message-Id: {MAILQUEUE-101.940413083221.320-at-parmly1.parmly.luc.edu} To: microscopy-at-anlemc.msd.anl.gov
} In answer to Phil Rutledge's query about fixation of "polysaccahridases": } The closest thing I know to a fixative for polysaccharides is Alcian Blue. } Jan Coetzee } Ruthenium red is used to demonstrate polysaccarhide coats of cyanobacteria. I don't have a reference to hand, but I believe a receipe is in Dykstra's & Bozzola & Russell's books? Phil Oshel
Hi Ian Mercury sources tend to be much less stable, and are not white enough to provide both wavelengths at comparable intensities for many dual excitation combinations. They can be used for some dual emmission work. We use the same instrument you are considering, and have added the OSP-3 illuminator and photmeter system for rapid spot measurements of ion [] and pH. The system works well, exposes cells to minimal photo-oxidizing irradiation during the measurements, and can be driven from pc-based software. Let me know if I can help
Message-Id: {199404122000543157-at-qmgate.secrc.abb.se} X-Mailer: InterCon Dispatcher/SMTP for QuickMail X-Priority: 4
Looking for someone that has been working with oxidation, hydriding and wear properties of surface coated or surface modified zirconium base alloys (like Zircaloy 2 or 4). The environment is water or steam at ]300!C. I am interested in all topics related to the subject; coating or surface modification methods ( how to get a coating without defects going through the coating?), corrosion tests, investigation of tested samples (LOM, SEM, TEM, SAM, XRD, electrochemical impedance spectroscopy.
Bengt Stridh ABB Corporate Research S-721 78 Vasteras Sweden E-mail: best-at-secrc.abb.se Fax: +46-21-13 41 00 Phone: +46-21-32 30 67
REGARDING Cerius Software Vendor According to a brochure picked up at the latest MRS meeting, the Cerius software is sold through: Molecular Simulations, 16 New England Executive Park, Burlington, MA 01803 ph: (617)-229-9800.
In reply to Charles Butterick's question about old osmium - try calling a vendor of osmium to ask their "proffesional" advice, such as EMS, or Stevens Metallurgical corp.
} Does anyone maintain a list of software suppliers for TEM } applications? I am interested in programs such as MacTempas, } Crystalkit, Desktop Microscopist, etc. Addresses, phone numbers, ftp } site information would all be very useful.
MacTempas and Crystalkit Total Resolution 20 Florida Street Berkeley, CA 94707 Attn: Roar Kilaas 510-527-9100 Fax 510-527-9151
Desktop microscopist Virtual Laboratories 37 Highland Court Ukiah, CA 95482 Attn: Eric Schlienger 707-462-8037
Crisp and ELD Calidris Manhemsvagen 4 S-191 45 Sollentuna Sweden Attn: Sven Hovmoller 46 8 625 00 41
Prism Signal Analytics 440 Maple Ave East, Suite 201 Vienna, VA 22180 Attn: Brian Culhane 703-281-2509
DigitalMicrograph Gatan Inc. 6678 Owens Drive Pleasanton, CA 94588 Attn: Sheri Kurland 510-463-0200
NCEMSS and Interface Tool Telnet from ORAC.LBL.GOV
TCBED from ASU
Image from MSA EMMPDL
That's my list of software sources.
Roseann Csencsits Electron Microscopy Center Argonne National Lab
} In reply to Charles Butterick's question about old osmium -- What exactly could go wrong with a sealed vial of the metal? If the vial seal had been breached, wouldn't the osmium vaporize? I know em makes a person really paranoid about materials, but is there any other reason not to use the stuff?
Tobias Baskin * TEMPORARILY C/O Peter Hepler * Biology Department University of Massachussetts * Amherst * Mass 01003 Tel: 413 - 545 - 2698 FAX 413 - 545 - 3243
In message Charles J. Butterick writes: } Greetings, } We just received 176 grams (ampules) of osmium tetroxide. The } ampules, we are told, were bought some time back in the 1960's from Fisher, } Englehard (in individual wooden cases!), National Lead, and MCB. The } integrity of the ampules seems good as evidenced by the translucent } greenish color of the osmium. However, the osmium seems to have coalesced } into a single mass. } Does anyone have any comments, warnings or advice? Considering that } this gift is worth over $6K, I'd like to be able to use it. } Charles,
We have heard your plea for help and are delighted to be able to respond with our assistance. In our lab, we have over 25 years experience of working with osmium tetroxide and biological systems. Just send us about 25 ampoules(grams), we will test the lot against biological systems we are currently working with, and report the results back to you.
-- Gib Ahlstrand 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
Sounds like the osmium has melted at some time and then reformed. After quizzing another EM veteran here, learned that such ampules should still be fine as long as they show that bright green color. Avoid using any vials showing black flecks in the coalesced osmium, as that's a sign of degradation. Similarly, when put into solution, good vials will give a bright yellow-green solution as usual; vials with black specks will probably give solutions with no color or the off-gray color of an old solution.
David Hall Dept. Neuroscience Albert Einstein Col. Med. Bronx, NY
We consider to purchase sample cutters for bulk materials and 3mm slices to prepare transverse TEM samples for HREM. I appreciate for your advice. Welcome companies to send us the price.
Sandy Burany Physics Department Simon Fraser University Burnaby, B.C. V5A 1S6 (604)291-4082 Fax (604) 291-3592 Email burany-at-sfu.ca
Message-Id: {MAILQUEUE-101.940414095946.705-at-prl.pulmonary.ubc.ca} To: microscopy-at-anlemc.msd.anl.gov
We are attempting to demonstrate peroxidase/myeloperoxidase activity in neutrophils in cytospin preparations by using DAB. We find it works well on fresh cytospins, but if cytospins are left for a few days we get no reaction. Any suggestions as to how best to maintain enzyme activity in stored cytospins? Mark Elliott
Re} Peroxidase & Cytospins Why not try fixing the cytospins with aldehyde to retain the peroxidase activity? Neither formaldehyde of gluteraldehyde will affect the enzyme activity but may inhibit proteolysis. The aldehydes will work best on cells that have not been allowed to dry out and will only continue to work if the cells remain hydrated. Leave them in buffer or, if formaldehyde-fixed, in formaldehyde. Glutaraldehyde fixation is only useful if you are using DAB. It will not be useful for fluorescent studies unless the autofluorescence is quenched with sodium borohydride.
Does anyone have experience with a good commercially available Monoclonal or polyclonal antibody against collagen I. Our experiments are performed with rabbits so a polyclonal made in rabbit is useless, or should be conjugated to something (biotin, dig.) Thanks in advance.
with likely the addition of a Hitachi in the next year. each is "Best" for us for what it does and the application for which it is used for.
Let's see what everyone has to say on this one. It should be interesting. I'll just sit back and "listen" for abit.
Remember try to be objective if you are going to respond. This is intentionally not a moderated list and I want to keep it that way.
I'm sure that the EM manufacturers who are also subscribers to this list will be interested in the responses. In that regard, I'll ask them (i.e. any manufacturers) not to respond to this question. If I see something which is not a fair discussion or an objective reply. I will cut the discussion off. If any manufacturer wants to complain please address all mail to me personnally and I will formulate any appropriate notice(s) to the listserver....
Nestor J. Zaluzec ANL EMCenter zaluzec-at-anlemc.msd.anl.gov
For reliability I have been most happy with my Hitachi H-7000 and S-4000. They serve about 99% of the needs we have in a multi-user facility, for mostly biological EM. There are a few times when I wish I had something else for very speialized work, but on the who;e these serve us well. My suspicion is that one could say the same for most of the major manufacturerers' products on the market today.
I agree with Nestor, the best microscope is dependent upon what you wish to do. Even in cameras, the Nikon is not ideal for all applications. However, to join in the foray: We love our Philips 400, 515, and CM-30s.
On Fri, 15 Apr 1994, Larry Hawkey wrote:
} } I have tried to send this message twice but is keeps getting bounced back } to me. } } Some one is asking me about my recommendations on which EM to } buy. So my question to all of you is this: } } Is there in the EM community a sense that there is one } Electron Microscope that is the best that money can buy? } } If some one was to ask about 35mm cameras for instance. I } think that the general consensus is that Nikon is ahead of } the pack. Minolta and Cannon are good and may be a good deal } for the price but if money is not the first concern Nikon is } the first pick. } } I use the JEOL 1200EX II. It is great. I have also used } several of the 100 incarnations. I used the Ziess EM 10 } about ten years ago but I have really only used JEOL for the } past ten years. In addition there seems to be a heavy } concentration of JEOLS in the Duke University community so I } feel that my impression that Ziess is nice but JEOL is best } may only reflect my small world. } } Try to put your biases aside and send me your thoughts. } } Larry Hawkey } Hawkey-at-neuro.duke.edu }
Some one is asking me about my recommendations on which EM to buy. So my question to all of you is this:
Is there in the EM community a sense that there is one Electron Microscope that is the best that money can buy...
Larry Hawkey Hawkey-at-neuro.duke.edu
I will start by saying I am not a TEM user and my answer will support this. I am a microprobe operator and an SEM operator. However, I have supervisory responsibility for support and operation of a lab that has a JEOL 100CX STEM, a JEOL 4000FX 400kV STEM, a Philips EM301 TEM in addition to 2 JEOL 35C SEM's, a JEOL 6400 SEM and a JEOL JXA-8600 EMPA so I am not insensitive to the needs of a TEM user. When we bought our most recent STEM, the 4000FX, we came down to a choice between Philips and JEOL. When we bought our most recent SEM, the 6400, we chose between Philips, CamScan, and JEOL. I would advise any one that in buying a piece of EM equipment there are three considerations: Performance, Cost, and Service. Of the three, cost is the lowest priority. Obviously, if you have limited funds, cost is a consideration up front (i.e. you can't get something for nothing). Once you purchase, what you paid is meaningless. The two remaining criteria will be with you for the life of the instrument. I will let the rest of the EM community steer you on the performance subject but I want to stress that efficent, prompt, compitent service is in my opion as important as performance. The best instrument, when inoperable, is not very useful. Waiting weeks for service is not pleasant, especially when deadlines come and go. Service personel that are not equipped or trained can lead to frustration. Personally, I will sacrifice a liitle performance if the competitor provides better service. Admitedly, I can do this because I do little work that "pushes the envelope". Mosty of my work is routine and not instrument limited. If you are beating back the frontiers of the science, then weigh performance heavily. But always keep in mind that the instrument must be well maintained if you expect to achieve optimum performance on demand. This takes good service.
Message-Id: {9404151852.AA01866-at-odin.morph.med.umich.edu} Errors-To: {dennis%odin-at-odin.morph.med.umich.edu} To: Jay Jerome {jjerome-at-isnet.is.wfu.edu} Cc: Larry Hawkey {hawkey-at-neuro.duke.edu} , Microscopy-at-anlemc.msd.anl.gov, dennis-at-med.umich.edu {Pine.3.89.9404151432.A9297-0100000-at-isnet.is.wfu.edu} "Larry Hawkey" {hawkey-at-neuro.duke.edu} , microscopy-at-anlemc.msd.anl.gov
Reply to: RE} } Who makes the best TEM? Just a note of clarification as to what the person who is looking for a TEM is going to use it for;
The person I am talking to is only interested in Routine Biological TEM, looking at ultra-structure (yes that is still done). Things like counting synapses or looking to see if the Mitochondria are screwed up. Also by best I am interested in compairing The preception that one company just carries a slightly but consistantly better line of EMs.
--------------------------------------
The problem with your question "Who makes the best TEM" is that you haven't specified what you wany to use it for. The best microscope for ultra-high resolution imaging is not necessarily the best for general teaching of students or the best for high performance microanalysis. You need also to factor in the service available. For example, how many engineers does a company have in your area, how many microscopes like the one you are thinking of buying does each engineer have responsibility for (put another way, how much experience does the engineer have on your instrument - a person can be very good, but if yours is the only TEM in his area he will have only limited experience with it). What kind of microscopy do you need to do - fast turn-round screening of biological samples or extensive analysis of materials samples, and so forth. There are good and bad points about all the major manufacturers, and probably each instrument from each maker is the best in its own niche. The analogy with the 35mm camera is a little over simplified.
On Fri, 15 Apr 1994, Nestor J. Zaluzec-ANL EMCenter wrote:
} } Talk about a loaded question. Which microscope is best? } } To state the obvious which I'm sure will be repeated at least } twice or three times in the repies to this question. } } ***It depends on what your trying to do!*** } } CTEM, STEM, AEM, CBED, XEDS, EELS, AES..... } and the list continues, choose your poison. } } At the ANL EMCenter we have: } } JEOL (100cx & 4000EXII) } Philips (EM420T & CM30T) } AEI (EM-7) } VG (603Z) } } with likely the addition of a Hitachi in the next year. } each is "Best" for us for what it does and the application } for which it is used for. } } Let's see what everyone has to say on this one. It should } be interesting. I'll just sit back and "listen" for abit. }
Well, I'm close to retirement so I'll stick my neck out. For biological applications I have used RCA, Siemens, Hitachi, Jeol, Zeiss, and Philips. For ease of use, quality of construction, resolution, and reliability I'll go with the Philips. My work has always been conventional biological TEM.
No, we have not tried cutting the fibers in to disks and perfroming a 3-D reconstruction. I have not heard of this previously applied to composite microstructures, have you? Can you tell me more about this methodology? Can you point me to specific papers in the literature? Is this similar to the type of 3-D reconstructions we read about being used in medical imaging? If so don't I need some features in the image to key off of?
Thanks for your help,
P. Joyce
Dear Peter,
If you consider biological systems to be composite microstructures, then, yes we have a lot of experience with this technique. As applied to your carbon-fiber system, there are several things to consider: 1) Is there enough contrast between the fibers and the matrix? 2) If not, is there a way to add contrast, such as by infiltrating the matrix? 3) Is the thickness of each slice limited by transmissivity or by overlap of features--in the HVEM a spec- imen of several microns can be penetrated, but after about one micron, typical biological specimens have so many features that these can no longer be untang- led? Another possibility if there is low contrast is to find an element in the matrix which is not found in the carbon, and to do element mapping--this is in- herently much lower resolution than imaging--or energy-filtered imaging looking at an edge or the low-loss region. In any event, I presume you have settled on appropriate conditions and obtained images which have then been scanned and converted into digital files. If the sections were thin enough so that the carbon fiber position and orienta- tion do not change appreciably from section to section, you can just stack the images from successive sections and process the resulting file as if it arose from a confocal microscope series or some such. There are many image proces- sing packages which will allow you to view the image volume from various angles with various features highlighted--you do have to tell the program how to iden- tify the features, but at worst, you can do this pixel-by-pixel if necessary. We use SEMPER on our confocal microscope to do this, and some in-house programs in combination with, I believe, MOVIE.BYU to do this for electron micrographs. If the sections can be thicker--e.g. if there are rather few carbon fibers distributed in a non-carbon-containing matrix and if energy-filtered imaging sees the carbon as dark while the matrix is transparent--you should either take stereo pairs and use a pointer to outline the features of interest within the section volume while viewing the stereo pairs, or take a large ser- ies of images at various tilts and do tomographic reconstruction. This may be more work than thin sections, but it has the advantage that features in the middles of sections are not perturbed by the cutting process--which can be a problem with thin sections. For reconstruction using stereo pairs, we use an in-house program, STERECON, and for tomagraphy, we use SPIDER, also an in-house program. SPIDER does reconstructions very much like medical imaging using either Fourier tech- niques, back-projection or projection onto convex sets. Fourier techniques are an old tried-and-true (mostly) method which allows filtration to smooth out noise, and which is well-understood, but which also gives artefacts due to the inevitable missing wedge of information. Back-projection just models the three dimensional object which would give rise to the observed projected views. Fin- ally, projection onto convex sets is a recent method incorporating constraints which, among other things, is used for de-blurring photographs. I don't under- stand this method well enough to try to explain it. Dr. Joachim Frank at our lab is in charge of the image processing end of things, Drs. Bruce McEwen and Michael Radermacher have experience with both programming and using SPIDER, Mr Mike Marko is our resident expert on STERECON and Mr. Ardean Lieth knows about all the programs. A letter (or e-letter) to any of these people should get you more information, and would be better than reading the papers cited for using these programs. I'm sure there are very good write-ups of serial-section techniques, but I am not familiar enough with the literature to reccommend any in particular. All of the programs developed here are available (I'm told the price is reasonable). Dr. Frank is the correct person to tell you about this aspect. I have probably told you more than you wanted to know, but I'd be happy to try to answer any other questions I can. The address for any of the people mentioned is Wadsworth Center for Labs. & Research Empire State Plaza, P.O. Box 509 Albany NY 12201-0509 and the e-mail addresses are username-at-tethys.ph.albany.edu, however, I don't know every one's usernames--try lastname, initial+lastname or initials as gues- ses. Good luck.
Your 'old' osmium should be ok! Concerning the fact that it coalesced into a mass on the bottom of the tube, this is not important: I remember a recipe (by S.Palay) recommending heating the tube (in a water bath) in order to collect all the osmium tetroxide in a single mass at one end of the tube. It worked ok (by the way, there is no point in doing so, if you carefully wash the outside of the ampulla before breaking it ;-)) Cheers! John
*********************************************************** * Jean C. Desclin (John), Associate Prof. of Histology * * Laboratory of Histology - Faculty of Medicine * * Brussels Free University (U.L.B.) * * e-mail: desclinj-at-ulb.ac.be (internet) * * snail mail: route de Lennik 808 * * B - 1070 Brussels Belgium * ***********************************************************
Reply_ RE} } General: Which Scope is Best? Reply from: John Mansfield North Campus Electron Microbeam Analysis Laboratory University of Michigan 2455 Hayward Ann Arbor MI 48109-2143 Phone: (313)936-3352 FAX: (313)936-3352 jfmjfm-at-umich.edu or John_Mansfield-at-mse.engin.umich.edu IMHO (In My Humble Opinion):
For quality of construction and reliability I would say Philips. In addition they have always been the best scope for CBED.
The one major thing that I dont like about ALL of the newer microscopes is the manufacturer's insistance in making the tilting and translation functions motorized. With these instruments you have no "feel" for the specimen position or tilt and when performing diffraction experiments I like to be able to "tweak" the tilts and sample position manually.
Our lab currently has 2 JEOLs, a 200cx and a 6100. Both instruments are under service contracts with JEOL, and I cannot imagine life otherwise. The contracts may seem expensive at first glance, but when a problem arises the service technicians have always fixed them much more quickley than we could have. The routine maintenance provided by the contracts also helps keep the instruments in good working order even with multiple users. If you have the option, life is much better witha service contract from the manufacturer.
John Phelps NIST, Boulder PHELPS-at-ENH.NIST.GOV
P.S. I promise that I wasn't paid by the service people for the response.
At Argonne National Laboratory, we have maintenance contracts on all of our TEM's, including an old JEOL 100CXII. We simply don't have the time to devote to repair work while the field service techs have the resources to take care of most of the problems that arise. We do some of the repairs ourselves, if they can be done quickly. For instance, we don't ask for help in changing filaments or in fixing a camera if the film gets jammed. We know our limits. One recent repair on our Philips EM420 took a week, and we could not have done it ourselves. Since we have a large base of users, 4 TEMs, 1 STEM, and only 3 staff members, time is very valuable to us.
Russell E. Cook Electron Microscopy Center for Materials Research Materials Science Division Argonne National Laboratory Argonne, IL 60439, USA
Dr. Bogers I talked with our ECM expert for advice on ab's and he suggested a company in Germany called HEYL Goerzallee 253, D-1000, Berlin 37 phone 030-8-17-60 52 FAX 030 8 17 40 49
He also mentionned a company called Collagen Corporation in Palo Alto, California USA, and one in Birmingham Alabama USA which also deals specifically with collagen antibodies. There is also a Developmental Biology Hybridoma Bank in the US which deals with ECM antibodies. He said that any of these may be able to help you and should be fairly good.
IMHO I would not suggest buying a JEOL 100B. This is the microscope that I never could qualify on at ASU, it took a lot of maintenance, and is 20 years obsolete. :)
I would also like to point out that if you want to unsubscribe you must send the message to the listserver, not the list:
listserver-at-anlemc.msd.anl.gov
We should all keep the information that was sent us when we first logged onto this list so that we know how to log off without annoying 1200 listmembers .
regards Mark W. Lund, PhD Director MOXTEK, Inc. Orem UT
I know we are a special case--we have a 1.2 MV HVEM from AEI--but we get along very well without a service contract (} 80% uptime). Our secret is that no ser- vice contracts being available on our beastie, we have had to learn to do it all ourselves. The service people in-house are VERY good--a must. Since we do a lot of development and improvement, we need a person who knows the machine very well, and those people also can service the microscope. If you have a fac-ility which just uses the microscope--as opposed to redesigning it--it is prob- ably just as well to have a service contract, but if you need a machine person or two on hand anyway, you can get along without a contract.
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I am sending this again because I am not sure if it was received properly the first time.
Dr. Bogers: I talked with our ECM expert for advice on ab's and he suggested the following companies as good sources. HEYL, Goerzallee 253, D-1000, Berlin 37, Germany Phone 030 8 17 60 52 FAX 030 8 17 40 49
also, a company called Collagen Corporation in Palo Alto California, USA and one in Birmingham, Alabama USA which also deals specifically with collagen antibodies. There is also a Developmental Biology Hybridoma Bank in the US which deals with ECM antibodies . He said that any of these may be able to help you and should be fairly good.
Message-Id: {1994Apr18.101055.761680489-at-ms.sjdccd.cc.ca.us} To: microscopy-at-anlemc.msd.anl.gov (microscopy list)
Dear fellow microscopists, At San Joaquin Delta College, Dept. of Microscopy, Stockton, CA we have an intensive 2 yr hands-on program to train microscopists. We issue certificates in both biological and materials area. We will be graduating 17 students from the program May 28, 1994. If you need an entry level person with 2 yrs intensive experience in TEM, SEM, EDS, OLM, all preps, routine maintenance, report writing, etc. etc. please contact Judy Murphy (murphy-at-ms.sjdccd.cc.ca.us, phone 209/474-5284; fax 209/474-5649) If you want more info about the program or a copy of the Delta Microscopy Newsletter, also contact as above. Our program has been in existence for 24 yrs. and we have graduates all over the United States and several abroad. Hope to hear from those interested. Judy Murphy, San Joaquin Delta College, Dept. of Microscopy, 5151 Pacific Ave., Stockton, CA 95207
JEOL 1200EX Purchased 1989, S/N EM157085-528 Under service contract since purchase Has Tracor Northern EDS (don't know model) Available immediately, shipping negotiable
For more information, contact: melilly-at-aol.com directly, not this list.
John chandler-at-lamar.ColoState.EDU Fort Collins, CO
Interesting and topical question. In my opinion unless inhouse expertise includes a trained microscope service engineer, you are playing russian roulette without a service contract. sooner or later the chamber is going to come up loaded. Over the long term it is not cost effective. However, I have often thought that facilities with good general in house expertise (we fix a lot of minor problems ourself) should be able to negotiate a different kind of contract (with different cost) than a facility that needs a service engineer for every minor problem. Perhaps we need a lobby with the microscope companies.
Having said that, and persuant to Larry Harkey's queries about microscopes,.o Our Philips service engineer in North Carolina is the best I have ever encountered and Philips in general has always provided superior service. However, with any company their are regional differences in the quality of service.
On Mon, 18 Apr 1994, Griffin, Robin wrote:
} } How are the majority of EM labs maintaining their electron microscopes? Are } they using maintenance contracts, in-house repair or something else? I } know how much maintenance contracts cost (EEK!). How well does it work } without maintenance contracts? I would appreciate any opinions about this } very expensive issue. }
How are the majority of EM labs maintaining their electron microscopes? Are they using maintenance contracts, in-house repair or something else? I know how much maintenance contracts cost (EEK!). How well does it work without maintenance contracts? I would appreciate any opinions about this very expensive issue.
How are the majority of EM labs maintaining their electron microscopes? Are they using maintenance contracts, in-house repair or something else? I know how much maintenance contracts cost (EEK!). How well does it work without maintenance contracts? I would appreciate any opinions about this very expensive issue.
Support person for the Electron Microscopy Core Laboratory of the Interdisciplinary Center for Biotechnology Research at the University of Florida.
DUTIES: Preparation and examination of biological specimens for transmission and scanning electron microscopy in a facility that serves the entire university community on a fee for service basis. Collaborative research possible as part of the program. Incumbent is expected to deal directly with the clients on experimental design, execution of the project and preparation of the data for publication, in consultation with the Scientific Director of the laboratory.
QUALIFICATIONS: B.S. or M.S. in a biological science with proven experience in most aspects of biological electron microscopy. Good communication skills, both oral and written. Ability to deal on a one to one basis with investigators from a wide variety of disciplines and backgrounds.
STARTING DATE: July 1, 1994. (Possibly sooner)
APPLICATION DEADLINE: Open until position is filled.
RANK: Appropriate to qualifications (EM Tech., Sr. EM Tech, EM Lab Manager)
SALARY: $17,873.-$22,633.
Inquiries by E-mail or full resumes to the address below
We currently have a Kodak slow-scan CCD system mounted on a 2010; this is a side-mount systems as opposed to the more common Gatan bottom-mount. Does anyone have any comments comparing the technical capabilities of these systems? How much does the placement affect usage, etc...?
Daniel L. Callahan Department of Mechanical Engg. and Materials Science Rice University dlc-at-owlnet.rice.edu
I'm trying for the first time to make some preparation of DNA for TEM. Grids and plates, with some initial problem, seem to be promising. My problem, now, is the elaboration of these pictures. I have seen that I can obtain good digital images with a scanner from the positives, and save them as TIFF files in a PC. I would like to find some PD program (possibly DOS, Windows or Amiga) to elaborate these files: the program should recognize DNA in the image and make some kind of elaboration: I don't know exactly what kind: I think (I hope!) I should be able to do something with these digitalised pictures.
Thank you for your help.
Nanni Iazzetti Dept. Genetics, General and Molecular Biology University of Naples Italy
Re} Best Microscopes I didn't see much use in joining the debate about "best microscopes" until the new data arrived. Buying a microscope for the US is a relatively easy choice and the discussion has covered all points. However, I lived and worked in Nairobi, Kenya for 7 years where we had a Zeiss EM 10A which worked well (and is still working). I think that this was the best choice for an EM in Africa. Although we only saw a service engineer once a year, the microscope was constructed for ease of service and repair. The machine was only used by a few people so a talented local electronics engineer and myself were able to keep this microscope running continually with relatively little effort. The long range support from Zeiss was exceptional and any serious problems were overcome with a few phone calls and faxes. Currently, I work on a Phillips 410 and Jeol 100cx, neither of which I feel capable of fixing when they go wrong. I could fix a Peugeot in Africa but have no idea on where to start with the computerized, fuel-injected cars over here.
Paul Webster Yale University School of Medicine 333 Cedar Street, New Haven, CT 06510.
Even more important, is how much knowledge the use (s) has. For stardard biological observations upt 50K even an old Zeiss S2 wil do the job. Of course, electron coherence, astigmatic correction, and vacum cleanness could drive you crazy, not to say much about frequent aligments. Thus, check your packet book, test your technicians, and determine what you need answered, then have all manufacturers to view and examine the same grid with the problem and make decision.
Cesar D. Fermin, Ph.D Tulane Medical School Pathology/SL79 New Orleans, La 70112
I agree with Nestor, the best microscope is dependent upon what you wish to do. Even in cameras, the Nikon is not ideal for all applications. However, to join in the foray: We love our Philips 400, 515, and CM-30s.
On Fri, 15 Apr 1994, Larry Hawkey wrote:
} } I have tried to send this message twice but is keeps getting bounced back } to me. } } Some one is asking me about my recommendations on which EM to } buy. So my question to all of you is this: } } Is there in the EM community a sense that there is one } Electron Microscope that is the best that money can buy? } } If some one was to ask about 35mm cameras for instance. I } think that the general consensus is that Nikon is ahead of } the pack. Minolta and Cannon are good and may be a good deal } for the price but if money is not the first concern Nikon is } the first pick. } } I use the JEOL 1200EX II. It is great. I have also used } several of the 100 incarnations. I used the Ziess EM 10 } about ten years ago but I have really only used JEOL for the } past ten years. In addition there seems to be a heavy } concentration of JEOLS in the Duke University community so I } feel that my impression that Ziess is nice but JEOL is best } may only reflect my small world. } } Try to put your biases aside and send me your thoughts. } } Larry Hawkey } Hawkey-at-neuro.duke.edu }
There are several short courses offered (1 week) through the Marine Biological Laboratory in Woods Hole, Massachusetts. For information call 508-548-3705 . The next one is in May. Regards, Nina Allen
On Tue, 19 Apr 1994 IDESOUSA-at-BRMVM1.VNET.IBM.COM wrote:
} Is anybody aware of a good short course on optical microscopy? } It is such a vast subject (polarized , UV,IR, confocal,filters, } image analysis, interferometry, ellipsometry etc.)
Might try the following:
1. McCrone Research Institute 2820 S. Michigan Ave. Chicago, IL 60616-3292 (312) 842-7100 [phone] 842-1078 [fax]
*some of their offerings: photomicrography, polarized light microscopy, hotstage microscopy, SEM, TEM, pharmaceutical, forensic, asbestos id, hair and fiber id, wood and pollen id, anb many others, including custom planned on-site courses. I've taken the Applied Polarized Light Microscopy course there, and found it -very- good; although not enough emphasis for materials type. Courses are spread out through the year. Prices for 1994: $1050 (ouch!) for almost all, w/ TEM being higher.
2. Institute for Microstructural Analysis c/o Buehler Ltd. P.O. Box 1 Lake Bluff, IL 60044 (702) 295-4659 [phone]
*courses offered include: image analysis, petrographic prep., basic metallography, advanced materials, ceramics, etc. Prices vary from $700 for 2-day to $1095 (again, ouch!) for 5-day, and higher prices at their Irvine, CA locale. I myself have not been there, but have heard good reports.
3. ASM International Materials Park, OH 44073 (216) 338-5151 [phone]
*courses include: metallographic interpretation, met. techniques, advanced tech. in optical (& electron) microscopy, failure analysis, optical microscopy of ceramics, quantitative metallography, etc... I've taken one course there, and have had several home-study classes. Good all around training. They offer what's called Materials Engineering Institute Extension diploma, and a three-tiered Center for Applied Metallography levels. Prices are similar to Buehler's.
} Could someone suggest books or reference material which is neither } trivial or too technically involved in the physics. (level of a good } metallurgical technician)
Some of my favorites include:
Metallography: Principles and Practice...........Vander Voort, George McGraw-Hill, 1984 (wish it would be updated tho) Polarized Light Microscopy.......................McCrone & Delly Ann Arbor Science Publishers, 1978 (likewise...) -and- many of the Microstructural Science volumes from the proceedings of the annual technical meetings of the International Metallographic Society. -and- many of the ore microscopy books have very detailed and highly useful information.
In addition, the other large met. supply houses besides Buehler, LECO [(616) 983-5531; St. Joseph, MI] and Struers [(216) 871-0071, Westlake, OH] offer tech. tips and misc. publications that are -very- handy and helpful. LECO also offers some training.
} One particular item I would find very handy is a compilation of } uv fluorescence wavelength of common substances. } } Thanks to all!
I'd be interested in such also...
Hope the above is helpful; glad to see another met online! -Rob ,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,, | Rob Tayloe | MSM Spelunkers Club /-v-\ | | Metallographic Lab. | Missouri Speleological Survey \-v-/ | | Rolla Research Center | Bat Conservation International \-v-/ | | U.S. Bureau of Mines | Missouri Cave & Karst Conservancy | | tayloe-at-rorc.usbm.gov | National Speleological Society #32993 /-v-\ | | (314) 364-3169 x247 | American Cave Conservation Association | ''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''
On Tue, 19 Apr 1994 IDESOUSA-at-BRMVM1.VNET.IBM.COM wrote:
} Is anybody aware of a good short course on optical microscopy? } It is such a vast subject (polarized , UV,IR, confocal,filters, } image analysis, interferometry, ellipsometry etc.)
Might try the following:
1. McCrone Research Institute 2820 S. Michigan Ave. Chicago, IL 60616-3292 (312) 842-7100 [phone] 842-1078 [fax]
*some of their offerings: photomicrography, polarized light microscopy, hotstage microscopy, SEM, TEM, pharmaceutical, forensic, asbestos id, hair and fiber id, wood and pollen id, anb many others, including custom planned on-site courses. I've taken the Applied Polarized Light Microscopy course there, and found it -very- good; although not enough emphasis for materials type. Courses are spread out through the year. Prices for 1994: $1050 (ouch!) for almost all, w/ TEM being higher.
2. Institute for Microstructural Analysis c/o Buehler Ltd. P.O. Box 1 Lake Bluff, IL 60044 (702) 295-4659 [phone]
*courses offered include: image analysis, petrographic prep., basic metallography, advanced materials, ceramics, etc. Prices vary from $700 for 2-day to $1095 (again, ouch!) for 5-day, and higher prices at their Irvine, CA locale. I myself have not been there, but have heard good reports.
3. ASM International Materials Park, OH 44073 (216) 338-5151 [phone]
*courses include: metallographic interpretation, met. techniques, advanced tech. in optical (& electron) microscopy, failure analysis, optical microscopy of ceramics, quantitative metallography, etc... I've taken one course there, and have had several home-study classes. Good all around training. They offer what's called Materials Engineering Institute Extension diploma, and a three-tiered Center for Applied Metallography levels. Prices are similar to Buehler's.
} Could someone suggest books or reference material which is neither } trivial or too technically involved in the physics. (level of a good } metallurgical technician)
Some of my favorites include:
Metallography: Principles and Practice...........Vander Voort, George McGraw-Hill, 1984 (wish it would be updated tho) Polarized Light Microscopy.......................McCrone & Delly Ann Arbor Science Publishers, 1978 (likewise...) -and- many of the Microstructural Science volumes from the proceedings of the annual technical meetings of the International Metallographic Society. -and- many of the ore microscopy books have very detailed and highly useful information.
In addition, the other large met. supply houses besides Buehler, LECO [(616) 983-5531; St. Joseph, MI] and Struers [(216) 871-0071, Westlake, OH] offer tech. tips and misc. publications that are -very- handy and helpful. LECO also offers some training.
} One particular item I would find very handy is a compilation of } uv fluorescence wavelength of common substances. } } Thanks to all!
I'd be interested in such also...
Hope the above is helpful; glad to see another met online! -Rob ,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,, | Rob Tayloe | MSM Spelunkers Club /-v-\ | | Metallographic Lab. | Missouri Speleological Survey \-v-/ | | Rolla Research Center | Bat Conservation International \-v-/ | | U.S. Bureau of Mines | Missouri Cave & Karst Conservancy | | tayloe-at-rorc.usbm.gov | National Speleological Society #32993 /-v-\ | | (314) 364-3169 x247 | American Cave Conservation Association | ''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''''
the McCrone Institute in chicago has a number of short courses on various aspects of light microscopy - including things that would probably be of interest to you such as polarization, dispersion staining, etc
there are several good, simple books around that are probably at the level you want - check out the Oxford series - I know thee is an flouresence & think there may also be a polarization book - they present the basic theory and are inexpensive - there is also a very practical-oriented book with a yellow and white paper cover whose exact and author escape me ( it is siiting on my bookshelf at home)
if you are not satisfied with your other responses send me a message & I will try to get back to you
We are thinking of purchasing a sapphire knife for our Vibratome so that we can cut high-quality sections. The knives are about $700. Being somewhat poor, we were wondering if the knife is a good investment. What has been people's experience with these knives? Do they cut reliable high-quality sections? Any problems with them? Thanks for any opinions.
-- ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ Nancy L Desmond, Ph.D. Department of Neurosurgery University of Virginia Health Sciences Center, Box 420 Charlottesville, VA 22908
We (actually Ed Lachica) have found it an absolute requirement for cutting unfixed chick embryos. Fixed post-hatch brains didn't seem to cut any better. But sectioning unfixed brain, or lightly fixed embryonic brain is helped tremendously.
Glen MacDonald Hearing Development Laboratories RL-30 University of Washington Seattle, WA 98195 (206)543-8360 glenmac-at-u.washington.edu
On Wed, 20 Apr 1994, Nancy L. Desmond wrote:
} We are thinking of purchasing a sapphire knife for our Vibratome } so that we can cut high-quality sections. The knives are about } $700. Being somewhat poor, we were wondering if the knife is a } good investment. What has been people's experience with these } knives? Do they cut reliable high-quality sections? Any } problems with them? Thanks for any opinions. } } -- } ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ } Nancy L Desmond, Ph.D. } Department of Neurosurgery } University of Virginia } Health Sciences Center, Box 420 } Charlottesville, VA 22908 } } 804.924.5607 (voice) } 804.982.3829 (fax) } ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ }
I am studying Aluminium alloys samples with TEM and especially two kinds of them: Al 3003 (0.12%Cu-1.2%Mn) AL 6061 (0.6%Si-0.27%Cu-1%Mg-0.2%Cr) I will use a double jet electropolisher (Tenupol 2) and I would want to know, if possible: 1-what are the best electropolishing solutions for these alloys (the books are not enough specific ) ? 2-what are the value of current,voltage and what is the temperature used for the solution ? 3-I suppose that we use the classical technique for cleaning but there must be some problems to prevent samples from oxydation which is an important problem for aluminium.Is there a special technique to avoid oxydation ?
NOTE:if no answer is possible, what are at least the best solution and electropolishing conditions for PURE aluminium.
It took nearly three months for BioRad to replace our laser when it failed after only 100 hours of use. So much for 24 hr. service. ********************************************************** * Greg Erdos ** * * Director, ICBR EMCL ** Phone 904-392-1295 * * 218 Carr Hall ** FAX 904-392-8598 * * University of Florida ** gwerdos-at-gnv.ifas.ufl.edu * * Gainesville, FL 32611 ** * **********************************************************
Does anybody know of a good procedure on the infiltration of yeast cells with LR-White? I dehydrate the pellet normally, starting with 35%ETOH for 10 minutes, 50%ETOH 10 minutes, 75%ETOH 10 minutes, 2 x 100%ETOH 10 minutes, 1:1- 100%ETOH/LR-white 1hour, 1:2- 100%ETOH/LR-White 1hour, 1:3- 100%ETOH/LR-white 1hour, 2x pure LR-white 1hour, bake at 50C for 72 hours. I am still ending up with holes around the cells and some cells aren't being infiltrated with the resin. I have never had problems with infiltration of any type of tissue, bacteria or anything else I've had to embed. This is getting to be pretty frustrating (Jack Daniels-black, here I come). Thanks Phil
Does anybody know the name of the journal that replaced "The Journal of Histochemistry and Cytochemistry" ? I was interested in getting a subscription to it and when I called Elsevier I was told it was discontinued 9 years ago. Someone said it was replaced by another outfit and they weren't sure who the publisher was. Anybody know? Thanks, Phil Rutledge
Message-Id: {MAILQUEUE-101.940422072110.367-at-prl.pulmonary.ubc.ca} To: microscopy-at-anlemc.msd.anl.gov
Phil Rutledge Our library carries a journal by the title of Journal of Histochemistry and Cytochemistry published by the Histochemical Society. It is printed by Williams and Wilkens in Baltimore. Probably is the same one you are looking for. In addition, concerning LR white and yeast infiltration, it was suggested by someone here that you might try using a mixture of osmium and potassium permanganate to fix the tissue, or possibly ruthenium red. You need to permeabilize the cell wall to let the LR white in. Try also to increase infiltration times, or put it under vacuum. Was also suggested you look up papers by C. Bracker on fixation of fungi. Hope this info helps.
I did the studies about Al3003 by TEM. You can find my paper "ATEM study of precipitates in an Al-Mn-Si alloy" publisned on METALLOGRAPHY 21:293-315 (1988). I used 20 HClO4 + 20 HCl + 80 ethyl alcohol, around 25 mA and temperature aroun -25 C. As you can see from my paper I have get lattice image. I am the first author of the paper XIANYING MENG-BURANY. Sandy X. Burany
Apologies to all concerned if this query is a bit dumb! After trawling the literature on HREM imaging theory (particularly the application of the wave-optical Abbe theory of image formation) I am confused as to whether the Fraunhofer diffraction pattern is:
i)a Fourier transform of the object exit wavefunction (being a product of the incident wave amplitude and a transmission function)
OR
ii)as some authors simply state, "the transmission function"
The J Histochem Cytochem is published by the Histochemical Society. The address is: Mount Sinai School of Medicine, 1 Gusatave L. Levy Place, Box 1045, New York NY 10029 Ph: 212-362-1801. You should find this journal in most university libraries. If your library does not get it, talk to your acquisition librarian. It is still the leading histochemical journal.
Denis G. Baskin, Ph.D. University of Washington
On Fri, 22 Apr 1994, rutledge phil wrote:
} Does anybody know the name of the journal that replaced "The Journal of } Histochemistry and Cytochemistry" ? I was interested in getting a } subscription to it and when I called Elsevier I was told it was } discontinued 9 years ago. Someone said it was replaced by another outfit } and they weren't sure who the publisher was. Anybody know? } Thanks, } Phil Rutledge }
} One particular item I would find very handy is a compilation of } uv fluorescence wavelength of common substances.
For ultraviolet(365nm) and blue-violet(400nm) fluorescence of over 600 materials see:
The Particle Atlas, Edition Two, Volume IV, McCrone and Delly (1979). [Out of Print]
The entire six-volume, electronic edition of the Particle Atlas is now available (PAE^2) on CD-ROM from McRI or:
Microdataware 2894 Tribune Ave Hayward CA 94542 TEL 510.582.6624 FAX 510.582.8295
McCrone Research Institute is offering the PAE^2 Version 1.0 for Windows with an educational discount of $550.00, which reduces the price to students to $950.00 if purchased within 90 days of course completion.
For a basic library of microscopy:
"A Basic Microscopy Library", John G. Delly, The Microscope, vol. 34, pp. 11-25 (1986).
This article contains a list and description of books on microscopy for the fields including biomedicine, mineralogy, petrology, petrography, metallography, chemical microscopy, and, of course, general works.
100%ETOH/LR-white 1hour, 2x pure LR-white 1hour, bake at 50C for 72 hours. I am still ending up with holes around the cells and some cells aren't
being infiltrated with the resin. I have never had problems with
infiltration of any type of tissue, bacteria or anything else I've had to
embed. This is getting to be pretty frustrating (Jack Daniels-black,
here I come). Thanks Phil
} } } } } } } } } } } } } } } } } } } } } } } } }
We routinely use LR White for preparing EM sections of yeast in immunolocalization experiments. The critical factor for getting good infiltration is a brief treatment of the fixed, washed cells with 1% sodium metaperiodate. This treatment alters the cell wall so that the resin infiltrates perfectly. After treatment with metaperiodate, a relatively standard dehydration/infiltration schedule can be followed. We also use a temp block (like the ones normally used for restriction digests) to precisely control temperature during polymerization. It seemed to be important to keep the temperature low during polymerization to minimize shrinkage (we used 45-50C for 24 hr).
For details of the procedure see:
Wright and Rine, Methods in Cell Biology 31:473-512.
van Tuinen and Riezman, J. Histochem. Cytochem. 35:327.
Phil wrote: ""Does anybody know of a good procedure on the infiltration of yeast cells with LR-White? I dehydrate the pellet normally, starting with 35%ETOH for 10 minutes, 50%ETOH 10 minutes, 75%ETOH 10 minutes, 2 x 100%ETOH 10 minutes, 1:1- 100%ETOH/LR-white 1hour, 1:2- 100%ETOH/LR-White 1hour, 1:3- 100%ETOH/LR-white 1hour, 2x pure LR-white 1hour, bake at 50C for 72 hours. I am still ending up with holes around the cells and some cells aren't being infiltrated with the resin.""
If you want to try and look at some of your poorly infiltrated yeats. If you have a 200KV instrument, you can cut 1/4 micron sections(maybe the yeast will stay in the plastic and not fall out!); stain with aqueous UA for 45'-1 hr. at 45 degrees, LC 30' and use the 200kv. We do this alot when people send us blocks that are bad.
This is what we do in our facility, with success on yeast: we use LR white, both medium and hard grades; for regular ultrastructure(medium grade)- if we have problems with infiltration: we increase the changes in LR white and make sure each change is accompanied by time on the rotator( -at- 4 RPM)(called rotamix in protocol below) we leave the sample in LR white overnight, on a rotator at 4 degrees. If we have trouble with air bubbles, we reduce stirring to a minimum and only use the rotator for mixing; we polymerize at 55 degrees for only 24 hours. For IEM, we use hard grade -if we have problems with infiltration: we increase the changes in LR white first and accept some yeast cells will not be infiltrated well, in some samples; we always have plenty in each section that are fine, even if we leave the cell wall intact. Note that there are two procedures; if your label is O.K. with procedure b, you'll get better infiltration; but either way gives use plenty of well-infiltrated yeasts cells in each section. If we have trouble with air bubbles, we reduce stirring to a minimum; we polymerize Hard grade at 50 degrees for only 24 hours.
Phil: here's the last half of our protocols; please call or send me e-mail anytime. It should work. Regular ultrastructure 6. Dehydrate : 30% - 5 mins 50% - 5mins 70% - 5mins 100% - three 5min. washes.
7. LR White:100% ETOH 1:1 - 30-mins RT on a gentle Rotamix*** .
8. LR White:100% ETOH 3:1 - 30-mins RT on a gentle Rotamix*** .
9. LR White - 2-3 changes 30-mins each, RT on a gentle Rotamix***
10. Overnight in fridge (-at-4oC), on a gentle Rotamix.
11. LR white - 1 change 1 hr (Warm LR White to RT before change)
12. Place tissue in gelatin capsules, add fresh LR White; seal capsules; allow to settle for 1 hour;
NOTE: if you need to centrifuge material and you have a small sample size, place cells in trimmed beam capsule, seal and centrifuge inside a 12-15ml cenrifuge tube{with a cotton bottom pad}, using a clinical centrifuge; then carefully seal beam capsule inside two 000 gelatin bottoms, that have been filled with LR White. Make sure you seal well with LR White. Use regular mold holders to hold capsules. OR if you need to centrifuge material and you have a large sample size, leave cells in original 00 gelatin capsule(i.e.complete step 12. Then place 00 capsule inside a BEEM capsule that has been cut to accomodate 00 gelatin capsule(i.e. top half is removed); centrifuge inside a 12-15ml cenrifuge tube{with a cotton bottom pad}, using a clinical centrifuge, until sample is spun down.
13. Polymerize at 55-60oC for 24 hours.
*** Use glass vials ; keep out of direct light; Use a rotomix, asLR White needs to be GENTLY stirred. KEEP Oxygen out of LR White; avoid excess air bubbles in resin. The dehydration and embedding are fairly straightforward. USE LR White Medium GRADE Open gelatin capsules to stop all polymerization.
For section stain, start with 2%UA aqueous for 20-45 mins. and LC 2-5 mins. try UA-45'/LC 2' first
FOR IEM Dehydrate : 50% - 30 mins 70% - 2 changes, 30 mins. each
LR White:70%ETOH - 2:1 2 changes 30 mins each ***
a. LR White - 3 changes 30-60 mins each, on a gentle Rotamix. b. LR White - 4-6 changes over three hours, on a gentle Rotamix.
a.Overnight in fridge (-at-4oC), on a gentle Rotamix. b. Place tissue in gelatin capsules, add fresh LR White; seal capsules; allow to settle. SEE NOTE step 8. Polymerize at 50oC for 24 hours.
a.LR white - 1 change 1 hour (Warm LR White to RT before change)
a.Place tissue in gelatin capsules, add fresh LR White; seal capsules; allow to settle for 1 hour;
NOTE: if you need to centrifuge material and you have a small sample size, place cells in trimmed beam capsule, seal and centrifuge inside a 12-15ml cenrifuge tube{with a cotton bottom pad}, using a clinical centrifuge; then carefully seal beam capsule inside two 000 gelatin bottoms, that have been filled with LR White. Make sure you seal well with LR White. Use regular mold holders to hold capsules. OR if you need to centrifuge material and you have a large sample size, leave cells in original 00 gelatin capsule(i.e. complete step 8). Then place 00 capsule inside a BEEM capsule that has been cut to accomodate 00 gelatin capsule(i.e. top half is removed); centrifuge inside a 12-15ml cenrifuge tube{with a cotton bottom pad}, using a clinical centrifuge, until sample is spun down.
9. a.Polymerize at 50oC for 24 hours.
TEST one block for sectioning quality. Polymerize another hour, if necessary.
*** Use glass vials to mix; mix solns. at RT; keep out of direct light; Use a rotomix, as solution is not very miscible & needs to be GENTLY stirred. KEEP Oxygen out of LR White; avoid excess air bubbles in resin. The dehydration and embedding are fairly straightforward.
LrWhite blocks should be stored at -20oC to preserve antigenicity indefinitely. Remove the gelatin and expose block to air in order to stop polymerization.
For section stain, start with 2%UA aqueous for 20-45 mins. and LC 2-5 mins. try UA-20' with no LC first, so you can locate the gold particles easily; adjust stain, as needed for ultrasruture.
JOURNAL OF MICROSCOPY - MAY 1994 ISSUE - VOLUME 174 PART 2
PUBLICATION DATE - 20 MAY 1994
Journal of Microscopy, Vol. 174, Pt. 2, May 1994, pp. 61-68
ACCURATE ALIGNMENT OF SETS OF IMAGES
By W. O. SAXTON Department of Materials Science and Metallurgy, Pembroke Street, Cambridge CB2 3QZ, U.K.
Summary Two modifications are described to the conventional procedure of cross-correlation, widely used for establishing the relative alignment of the members of a set of images from which a higher resolution or more interpretable restoration is sought. Both achieve a high and sharp peak in circumstances where the conventional peak is too ill defined to be recognisable; neither involves significant additional computational time. The more general method requires a rough knowledge of the imaging conditions, but a variant applicable to images with axial resolution has no such requirement. In addition, a least-squares procedure is presented for achieving an optimum compromise between many pair-wise displacement measurements, preventing the accumulation of alignment errors across a set of images.
Journal of Microscopy, Vol. 174, Pt. 2, May 1994, pp. 69-73
RESOLUTION IN NONLINEAR LASER SCANNING MICROSCOPY
By J. DEITCHE, M. KEMPE & W. RUDOLPH The University of New Mexico, Department of Physics and Astronomy, Albuquerque, NM 87131, U.S.A.
Summary The lateral and depth resolution of nonlinear microscopy was studied systematically. Nonlinear microscopy can be classified into several categories depending on the coherence properties of the process that generates the imaging signal from the illuminating light, on whether a single- or a two-beam geometry is used and whether the optical setup is Type I or Type II. An evaluation of the imaging equations shows that (i) lateral and depth resolution improve with increasing nonlinearity, (ii) the differences between coherent and incoherent imaging diminish, and (iii) nonlinear imaging allows depth discrimination in Type I microscopy.
Journal of Microscopy, Vol. 174, Pt. 2, May 1994, pp. 75-84
CRYO AUTOMATED ELECTRON TOMOGRAPHY: TOWARDS HIGH-RESOLUTION RECONSTRUCTION OF PLASTIC-EMBEDDED STRUCTURES
By M. B. BRAUNFELD, A. J. KOSTER, J. W. SEDAT & D. A. AGARD The Howard Hughes Medical Institute and the Department of Biochemistry and Biophysics, University of California at San Francisco, San Francisco, CA 94143-0448, U.S.A.
Summary The use of fully automated data collection methods for electron beam tomography allows a substantial reduction in beam dose. The goal has been to develop new protocols for data collection defining optimal approaches for maintaining data self-consistency and maximizing the useful resolution of the reconstruction. The effects of irradiation and post-cure microwaving were examined for a variety of embedding media (Epon, Epox, Lowicryl) in order to quantify beam damage with the goal of identifying the most beam stable embedding medium. Surprisingly, the substantial dose reduction made possible by automated data collection did not result in a significant decrease in specimen shrinkage even for samples stabilized by pre-irradiation. The accelerated shrinkage is a direct consequence of the stroboscopic illumination patterns inherent to automated data collection. Furthermore neither the choice of embedding resin nor microwave post-curing greatly affected shrinkage. Finally, cryogenic data collection was investigated as a means to minimize the effects of secondary radiation damage. Minimal pre-irradiation coupled with low-temperature automated data collection greatly reduces shrinkage and should result in high-quality data for three-dimensional reconstructions.
Journal of Microscopy, Vol. 174, Pt. 2, May 1994, pp. 85-92
A NOVEL METHOD FOR MEAN CELL VOLUME ESTIMATION
By P. WEBSTER* & G. GRIFFITHS+ *Yale University School of Medicine, Department of Cell Biology, 333 Cedar Street, New Haven, CT 06510, U.S.A. +European Molecular Biology Laboratory, Postfach 10.22.09, 69012 Heidelberg, Germany
Summary A novel method is described for the estimation of the mean cell volume of cell populations grown in suspension. The cells are filtered on to a nitrocellulose filter to form a cylindrical pellet which is embedded in epoxy resin. Using estimates of pellet height and radius, the number of cells in the pellet and the volume density of cells in the pellet, it is possible to produce an unbiased estimate of the mean cell volume. This method is compared, using cell suspensions of the blood parasite Trypanosoma brucei, with other published methods for mean cell volume estimation. A Coulter channelizer was also used to compare the mean cell volume of living trypanosomes with that of aldehyde-fixed populations, and the values obtained were compared with those obtained with the new method. The estimated mean cell volume of a T. brucei clone was used to derive values from volume densities obtained by point and intersection counts for the absolute volumes of the flagellar pocket, the nucleus and the endocytic organelles containing internalized horseradish peroxidase and transferrin-gold after 30-min incubations at 310K. Estimated values for the cell were also obtained. From known data on the total amount of variant surface glycoprotein molecules per cell and the known packing density of membrane proteins, it is estimated that approximately 80% of the molecules must reside in intracellular compartments. It was estimated that the equivalent of 5% of the surface membrane may be internalized per minute, an amount which is almost the size of the entire flagellar pocket membrane.
Journal of Microscopy, Vol. 174, Pt. 2, May 1994, pp. 93-100
THE INFLUENCE OF TISSUE PROCESSING ON QUANTITATIVE HISTOPATHOLOGY IN BREAST CANCER
By M. LADEKARL Stereological Research Laboratory, University Institute of Pathology and Second University Clinic of Internal Medicine, Institute of Clinical Experimental Research, University of Aarhus, Denmark.
Summary Objective grading of breast cancer by morphometry has been suggested for improving the precision of the prognostic prediction. However, the tissue components evaluated might be influenced by variations in the processing, reducing the clinical value. In the present study, the impact of the period of fixation, of the acidity of the fixative and of the embedding medium was investigated by allocating tissue samples from 27 surgical breast cancer specimens systematically randomly to different modes of processing. The volume-weighted mean volume of cancer call nuclei was estimated using the method of point-sampled intercepts on vertical sections. In addition, estimates of the mean nuclear profile area, the nuclear volume fraction, the nuclear profile density and the mitotic profile frequency were obtained. The quantitative histopathological estimates were stable with respect to the investigated variables of the tissue processing. No significant differences were found when comparing the estimates obtained in samples from five tumours fixed in formalin at pH 5.0, 6.0, 7.0, 7.4 and 8.0 respectively. Similarly, no significant correlations between estimates and the period of formalin fixation (24h, 3 days and 3 months) were found in samples from five other tumours. However, the volume-weighted mean nuclear volume of cancer cell nuclei was 13% larger (2p=0.004) and the mean nuclear profile density was 17% smaller (2p=0.04) in hydroxyethyl- methacrylate-embedded samples from 17 tumours as compared to paraffin-embedded samples. Thus, the shrinkage observed in paraffin seems to affect nuclei less than tissue.
Journal of Microscopy, Vol. 174, Pt. 2, May 1994, pp. 101-110
METHOD FOR THE STUDY OF THE THREE-DIMENSIONAL ORIENTATION OF THE NUCLEI OF MYOCARDIAL CELLS IN FETAL HUMAN HEART BY MEANS IF CONFOCAL SCANNING LASER MICROSCOPY
By Y. USSON,* F. PARAZZA, * P.-S. JOUK + & G. MICHALOWICZ+ *Dynamique de l'organisation du genome, Universite Joseph Fourier, BP53, 38041 Grenoble cedex 9, France +Medecine Neonatale, Centre Hospitalier Regional Universitaire, Grenoble, France
A series of three-dimensional image analysis tools are used to measure the three-dimensional orientation of nuclei of myocardial cells. Confocal scanning laser microscopy makes it possible to acquire series of sections up to 100 micrometre inside thick tissue sections. A mean orientation vector of unit length is calculated for each segmented nucleus. The global orientation statistics are obtained by calculating the vectorial sum of the nuclear unit vectors. The final orientation statistics are obtained by calculating the vectorial sum of the nuclear unit vectors. The final orientation is expressed by a mean azimuth angle, an elevation angle and a measure of the angular homogeneity. The method is illustrated for two different regions of the myocardium (interventricular septum and papillary muscle) of a normal human fetal heart. This quantitative method will be used to assess and calibrate the information provided by polarized light microscopy.
Journal of Microscopy, Vol. 174, Pt. 2, May 1994, pp. 111-119
A NOVEL APPLICATION OF MICROSPHERE PERFUSION AND SCANNING ELECTRON MICROSCOPY TO THE IDENTIFICATION OF PULMONARY ARTERIOLES IN GUINEA PIG AND RABBIT LUNGS
By D. C. WALKER, S. HOSFORD & A. MACKENZIE Christmas Seals Electron Microscopy Laboratory of the Pulmonary Research Laboratory, U.B.C. Pathology, St Paul's Hospital, Vancouver, B.C., Canada V6Z 1Y6
Summary In arterioles of the lung the intravascular blood pressures are lower than in comparable vessels in the systemic circulation and the arteriole walls are thinner. Therefore, it is very difficult to distinguish between arterioles and venules of the same size using scanning electron microscopy. This study describes a novel application of latex microsphere perfusion and scanning electron microscopy which distinguishes between pulmonary arterioles and venules on the basis of endothelial cell morphology. Microspheres, 90 and 45 micrometres in diameter, were perfused into the arterial side of the pulmonary circulation of guinea-pig and rabbit lungs. Scanning electron microscopy of the arterioles on both sides of the lodged microspheres indicated that the endothelial cells are spindle shaped. In contrast, the endothelial cells of equal diameter venules are polygonal. Furthermore, the nuclei of the arteriolar endothelial cells were significantly (P=0.019) narrower than those of endothelial cells in venules of equal diameter. Finally, it was observed that the differences between arteriole and venule endothelial cells persisted distally to the capillaries.
Journal of Microscopy, Vol. 174, Pt. 2, May 1994, pp. 121-123
ASTIGMATISM OF A THICK CYLINDRICAL OBJECT IN REFLECTIVE MODE CSLM
By CHANG-GUI WANG, H. RAIKKONEN & M. LUUKKALA Department of Physics, PO Box 9, 00014 University of Helsinki, Finland
Summary The axial response of a basic confocal microscope is determined when the sample is a thick cylindrical or tubular structure. The response from the back wall of the cylindrical sample is split into two separate signals due to basic aberration or astigmatism effects.
Journal of Microscopy, Vol. 174, Pt. 2, May 1994, pp. 125-127
Short Technical Note SEVERE SHRINKAGE OF MICROFIL DURING TISSUE CLEARING WITH THE SPALTEHOLZ TECHNIQUE
By J. MOLLER, K. ROBERTSEN & E. S. HANSEN Institute of Experimental Clinical Research, Department of Orthopaedics, University of Aarhus, Randersvej 1, DK 8200 Aarhus N, Denmark
Summary The effect of two commonly used tissue clearing methods on the morphology of vascular casts by Microfil, a silicone rubber injection compound, was investigated. Microfil undergoes extreme shrinkage when the casted tissue is cleared by the alcohol-methyl salicylate clearing technique. No shrinkage is observed when the alternative glycerine clearing method is used. The alcohol-methyl salicylate clearing technique should be avoided in studies employing Microfil.
PLEASE SEND YOUR QUERIES CONCERNING THE JOURNAL TO RMS-at-VAX.OX.AC.UK. WE ARE ALSO PLEASED TO ACCEPT LETTERS TO THE EDITORS BY EMAIL.
Does anybody know where I could find SJ Singer, the man who introduced EM immunocytochemistry by a paper in Nature (London) 1959. I would like to present him by his portrait at a meeting. All advices are welcome. Thanks, Sverker Enestr|m
Has anyone worked with a type of species in the bacteria field known as a marine cytophage type? I'm trying to fix the cells on agar and the cells keep floating up off the surface of the agar. I would like them to stay put. (maybe I need to put them in a training school for bacteria!). I've tried several methods suggested by members of the microscopy board but no success. Hopefully this problem can be taken care of. It's driving me nuts! Thanks, Phil
Message-Id: {9404251801.AA02278-at-odin.morph.med.umich.edu} Errors-To: {dennis%odin-at-odin.morph.med.umich.edu} To: microscopy-at-anlemc.msd.anl.gov Cc: dennis-at-odin.morph.med.umich.edu
Like a dumbbell I forgot to mention I need to do SEM on these little suckers. Fixing them and processing for TEM is no problem. That's easy. Trying to keep the little creatures attached to the agar surface and fixing them without them floating off is a #!$#%$$^&(^*& of a problem. I need to see the surface of the bacteria and how they are oriented in their growth on the agar without losing ANY of the cells. I should have become a mechanic or plumber 27 years ago, EM can be a real -at-$%^%& ! Thanx, Phil
Have you tried attaching them to a millipore filter by passing them from a syringe onto the filter (using millipore syringe fittings) and then fixing them onto the millipore surface by squirting fixative through the filter? This works for suspension culture cells and many small critters. The millipore filters can then be CPD and attached to a stub.
On Mon, 25 Apr 1994, rutledge phil wrote:
} Like a dumbbell I forgot to mention I need to do SEM on these little } suckers. Fixing them and processing for TEM is no problem. That's easy. } Trying to keep the little creatures attached to the agar surface and } fixing them without them floating off is a #!$#%$$^&(^*& of a problem. I } need to see the surface of the bacteria and how they are oriented in } their growth on the agar without losing ANY of the cells. } I should have become a mechanic or plumber 27 years ago, EM can be a real } -at-$%^%& ! } Thanx, } Phil }
Try this sandwich method - it works great for suspensions:
use the thinest layer of agar that you can remove with the colonies intact
- take a 13- 15 mm dameter nylon washer (you can use brass if you are not going to use OsO4) - I suppose you can use a larger diameter washer & filter so long as it will fit in your cpd unit - place an appropriate pore size filter on the washer - place a "spacer" on the filter (for cell suspensions I punch a hole in the blue spacer papres packed with the filters - you might want to try an o-ring) - cover with a second filter - top off the sandwich with another washer
clip the whole thing together with a paper clip (yours may be a triffle thick for that, so you may have to be inventive
we have used relatively short fix/wash/dehydrate times for suspensions with good results
the only catch is taht some of your cell may stick to the top filter (ours do) so you might need to make sure that "chamber" height exceeds the height of your sample and try to keep it right side up during processing
(an easy way to remove small sections of colonies from plates is with a cork borer or a glass pipette)
good luck & please excuse the typos
feel free to contact me if this is unclear
Marcelle A Gillott Univ Wisconsin-Milwaukee Department of Biological Sciences 414-229-4186
First: Thanks to all for their suggestions. Now: Let me go into this problem a little deeper. We have unknown bacteria of marine cytophage species that autofluoresce red and green. We want to look at the surface by SEM to see: 1: surface structures, if any 2: orientation of the bacteria as it grows (there is a definite orientation as seen after some of the cells have floated off) Need: a way to fix the cells without ANY cells floating off the surface Tried: 1: suspensions 2: growing on nucleopore filters 3: growing on coverslips Result: no autofluorescence, no definite orientation Fix: 1: direct addition of glutaraldehyde 2: cutting out pathways in the agar and fixing by letting the agar absorb the fix and fixing the cells from underneath the agar. 3: osmium vapor fixation then glut. Result: Some of the bacteria still float up when washing and dehydrating. Question: Is there a way to crosslink the cells by some fixation technique so they will remain in place on the agar surface?
John Mansfield North Campus Electron Microbeam Analysis Lab University of Michigan 2455 Hayward Ann Arbor MI 48109-2143 Phone: (313)936-3352 FAX (313)936-3352 jfmjfm-at-engin.umich.edu or John.F.Mansfield-at-umich.edu Time: 08:59
Date:4/26/94 NC EMAL
Macro 'Read AFM File'; {reads header in and searches for the number of Samps/line, uses this value} {as the width and heigth for the image import (therefore supports 128,256 and} {512 squared images} var size, width, height, offset, slices, first, second, third, fourth: integer; icount: integer; temp, one, two, three, four: string; done: boolean; begin; done:= 'false'; width:=8192; height:=1; Offset:=0; slices:=1; SetImport('8-bit'); SetCustom(width, height, offset,slices); Import(''); icount:=1; repeat temp:=chr(getpixel(icount, 0)); if ((temp='S') and (one {} 'S')) then one:=temp; if ( (temp='a') and (one='S') and (two {} 'a')) then two:=temp; if( (temp='m') and (one='S') and (two='a')) then three:=temp; if( (temp='p') and (one='S') and (two='a') and (three='m')) then four:=temp; if (four='p') then done:='true'; icount:=icount+1; until done='true'; first:=(getpixel((icount+8), 0))-48; second:=(getpixel((icount+9), 0))-48; third:=(getpixel((icount+10), 0))-48; fourth:=(getpixel((icount+11), 0))-48; if (first {} 1) then size:=(first*100)+(second*10)+third; if (first=1) then size:=(first*1000)+(second*100)+(third*100)+fourth; Dispose; width:=size; height:=size; offset:=8192; SetImport('16-bit signed,swap bytes'); SetCustom(width, height, Offset); Import(''); end;
Phil, Can you overlay the agar/bacteria with another thin layer of agar then fix as usual. When osmium is added the bugs should turn black (if in colonies) making them visible after emebedding.
Royal Microscopical Society & Oxford Brookes University
26 - 31 March 1995
Location Oxford Brookes University - Oxford's new University, situated 1 mile from the centre of historic Oxford. Full board accommodation will be available at Morrell Hall, 5 minutes walk from the campus.
Cost Registration and full board accommodation will cost no more than 440.0 pounds sterling. Receptions, the Conference Dinner and a copy of the special edition of the Journal of Microscopy are included in the cost. There are a very limited number of double rooms available.
Delegates To preserve the informal nature of the Botanical Microscopy Meeting, the number of delegates will be restricted to 150. Places at the conference will be allocated on a first come, first served basis with a preference given to those offering to present a poster. (Please note that places will only be guaranteed after payment has been received.)
Booking forms and abstract instructions will be available from June 1994.
Organizing Committee Nick Harris - Durham Chris Hawes - Oxford Nick Read - Edinburgh Peter Shaw - John Innes Institute
Format of Meeting Following the success of Botanical Microscopy 1991 in Durham, the same team are organizing the meeting to be held in Oxford. The scientific programme will be based around presentations from keynote speakers.
A call for abstracts will be made late 1994 and the organizing committee will select suitable papers from these and invite the authors to present them orally. However, posters on any aspect of plant cell biology involving microscopy may be presented.
Social A reception is planned for the opening evening of the meeting and an informal Conference Dinner will be held on the Thursday evening. A number of short tours around Oxford and its environs will be offered on the Wednesday afternoon of the Conference.
Publications Keynote and selected papers will be published as a special edition of the Journal of Microscopy which will be distributed to all delegates upon publication. All manuscripts will be subject to peer review.
Travel Air: Oxford Brookes University is a 60 minute bus ride from Heathrow airport and 130 minute ride from Gatwick airport.
Road: Coaches run from Victoria Bus Station in London every 20-30 minutes. Journey time is approximately 60 minutes.
Rail: There is a good train service from Paddington Station, London to Oxford.
Queries If you would like any further information, please contact Miss Karen Hale at the Royal Microscopical Society, 37/38 St Clements, Oxford, OX4 1AJ. Tel: +44-865-248768. Fax: +44-865-791237. Email: rms-at-vax.ox.ac.uk.
Programme to Include: žMicrotubule and cytoskeletal dynamics
žMicroscopy of living cells - ion imaging, cell-cell signalling
žPlant cell organization - cell walls, meiosis, low temperature techniques
žMolecular mechanisms of plant development - cell cycle, floral development
žPlant microbe-interactions
Keynote Speakers B Gunning - Canberra H Shibaoka - Osaka J Hush - Sydney S Giroy - Penn State K Oparka - Invergowrie K Roberts - Norwich M Parthasarathy - Cornell Z Cande - Berkeley J Doonan - Norwich R Howard - Wilmington
Please complete this coupon indicating your requirements, and return to the Royal Microscopical Society, 37/38 St Clements, Oxford, OX4 1AJ, UK.
I would be interested in attending the 5th International
Does anybody know where I could find SJ Singer, the man who introduced EM immunocytochemistry by a paper in Nature (London) 1959. I would like to present him by his portrait at a meeting. All advices are welcome. Thanks, Sverker Enestr|m
======================================================================= I send this mail again because of local configuration error: "Service unavailable" =======================================================================
Hi, We are considering the purchase of an Energy Dispersive X-ray Analysis (EDX) system for attachment to our SEM within the next year. Our applications are in two areas; paper making and corrosion of materials.
I would like to know if anyone is aware of mailing lists etc., accessable on the Internet, which are specifically interested in EDX. I would like to follow discussions on relative merrits of detector and window types , vacuum requirements for windowless detectors, pulse processor requirements, software options and in particular, read any horror stories associated with a given manufacturer.
If anyone has opinions on what is currently available from recent purchase deliberations or would like to recommend a review paper on current technology or a periodical which they go to for this sort of information it would be greatly appreciated.
My first priority is always dependable service, but having said that, I will be looking for a high quality detector capable of light element analysis, high pulse throughput, good quantitation with ease of use , beam control and image acquisition and replay capabilities.
Thanks for your help.
______________________________________________________________________ Laurie Frederick, A.SC.T. PAPRICAN Corrosion Control Group 3800 Wesbrook Mall The Pulp and Paper Research Vancouver, B.C. Institute of Canada Canada V6S 2L9
I am looking for some help regarding pore sizes in the secondary cell wall of wood. The wood has been partially degraded by fungi, and I want to see how the porosity of the lignin-cellulose substrate has changed. I plan on infiltrating wood with proteins of different molecular weights (my probes), then locating them with immunolabling. I have already decided on a 40000 M.W. (ovalbumin, and MN dependant peroxidase), and a 20000 M.W. (myoglobin) probe. I am still looking for an adequate probe for M.W. 5000, 10000, and 15000. By adequate probe I mean a protein that does not denature easily, and also I can purchase antibodies against. Any help would be appreciated.
Eugene Krueger, Dept of Plant Pathology, University of Minnesota.
-- Eugene Krueger 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
To: MICROSCO:CENTRAL cc: DRStadden:R_D:Armstrong Subj: Spencer Stereoscope ------------------------------------------------------------------
Is anyone acquainted with a good repair service for AO Spencer stereomicroscopes? We have the "gray standard issue", step magnification, ubiquitous model, probably vintage 1959 or so. One of the prisms has come loose, and we've spent too much time already trying to get it EXACTLY back into position. Maybe there's a trick or tool we're missing. Thanks in advance for any help.
I am looking for some help regarding pore sizes in the secondary cell wall of wood. The wood has been partially degraded by fungi, and I want to see how the porosity of the lignin-cellulose substrate has changed. I plan on infiltrating wood with proteins of different molecular weights (my probes), then locating them with immunolabling. I have already decided on a 40000 M.W. (ovalbumin, and MN dependant peroxidase), and a 20000 M.W. (myoglobin) probe. I am still looking for an adequate probe for M.W. 5000, 10000, and 15000. By adequate probe I mean a protein that does not denature easily, and also I can purchase antibodies against. Any help would be appreciated.
Eugene Krueger, Dept of Plant Pathology, University of Minnesota.
-- Gib Ahlstrand 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 question: To detect dim backscattered electron diffraction patterns, one generally uses a SIT (Silicon Intesified Tube) camera behind a phosphor screen. Hamamatsu's model of SIT has a quoted sensitivity limit of 10^-4 lux. Hamamatsu also sells (for a modest premium) an "intensified" CCD which is sensitive down to 10^-5 lux.
We know a SIT camera will work, but imagine the CCD system will work a little better.
Has anyone used an intensified CCD (Either Hamamatsu's or others')? Are there any drawbacks besides the price? Are there better SIT systems available that would rival the intensified CCD?
Thank you, Chris Krenn Graduate Student UC Berkeley Dept. of Materials Science
Are you working with tissue or purified amyloid? If tissue you need to permeablize the tissue with a detergent (such as Triton-X 100). It seems accesability is your problem. Try various concentrations of detergent, 0.1 to 1.0% for various times. Detergents are used after fixation and in all blocking, Ab's, and probe solutions. Hope this helps.
Wayne England has asked: *********************************************************************** I am having a problem getting Unicryl to polymerize properly and am left with a very brittle block or no polymerization at all. I have tried all of the manufacturer's methods (heat and UV covering all the variables) but am not satisfied with the results. Do you find this resin to be normally brittle or am I missing something? Also, does anyone know how this resin is at infiltrating plant material? It's properties sound too good to be true!! *********************************************************************** We have some experience using this polar resin, which has some ad- vantages over Lowicryl K4M but shares the general disadvantages inherent in hydrophilic resins. The labeling intensity is high but more varying than for Epon. I polymerize at -10C by direct UV in the low temperature chamber of Balzers FSU 010 freeze substitution unit for at least 2 days and using the low temperature UV polymerization insert, filled with 10 ml ethanol. The blocks become hard rubber-like and are more easily sectioned than Lowicryl. The ultrathin sections are more resistant to the electron beam than Lowicryl but not as good as Epon. The tecnicians are anxious about hypersensitivity reactions after exposure for some time. What are yours opinion? Sverker Enestr|m Dep of pathology Link|ping, Sweden
Brij 58 is another detergent that works well for immunocytochemistry. It is not as harsh as Triton and preserves a little more ultrastructure. Downside is that it often doesnt permeabilize all cells in a single preparation.
On Wed, 27 Apr 1994 EMLAB-at-opus.mco.edu wrote:
} Monica, } } Are you working with tissue or purified amyloid? If tissue you need to } permeablize the tissue with a detergent (such as Triton-X 100). It seems } accesability is your problem. Try various concentrations of detergent, } 0.1 to 1.0% for various times. Detergents are used after fixation and in all } blocking, Ab's, and probe solutions. Hope this helps. } } Ed Calomeni }
Message-Id: {9404271519.AA00742-at-odin.morph.med.umich.edu} Errors-To: {dennis%odin-at-odin.morph.med.umich.edu} To: microscopy-at-anlemc.msd.anl.gov Cc: dennis-at-odin.morph.med.umich.edu
One of my library patrons is studying corrosion of metal by bacteria. She needs techniques for preparing samples of bacteria on metal for SEM microscopy. She is also looking for methods of staining or gold-labelling to improve SEM sensitivity. The methods need to be applicable to a wide variety of bacteria species. Any information on best SEM conditions - voltage, angle, windows, etc. would also be helpful.
Thanks for your help.
Replies can be made directly to me at smiths-at-mlc.lib.mi.us or you can contact my patron, Cathy Stewart at 313/676-5292.
Susan Smith R&D Tech. Information Center National Steel Corp. Trenton, MI 48183
Message-Id: {MAILQUEUE-101.940427100951.630-at-prl.pulmonary.ubc.ca} To: microscopy-at-anlemc.msd.anl.gov
We inherited a microscope slide awhile back which we would like to replace, but do not know of a source. The slide had a grid of letters on it (black background with white letters) and it was used for finding particular locations of structures on other slides; ie a certain cell on your slide was at position Xy on the reference slide. Ours is getting so beat up that it is going to fall apart any day. I am not sure if it was commercially available, or made by someone. Any help would be appreciated.
Mark Elliott UBC-Pulmonary Research Lab St Pauls Hospital Vancouver
We currently use Agfa Scientia film for (300kV) HREM. It is a little slower (maybe) than Kodak SO163, faster than 4489. It has a slightly finer grain than Kodak SO163, which helps. Two questions: 1) What are people using for faster film. The Agfa film is quite old (i.e. it has been around for at least ten years), and is there anything better at a reasonable price? 2) We have noticed some scratches at times which seem to be a quality control issue in the film. Is this common?
Scanning: the journal of scanning microscopy is one journal that you may want to check. It is published by FAMS Inc., Box 832, Mahwah, NJ 07430-0832 Phone 201/818-1010 FAX: 210/818-0086 Hope this is helpful.
Susan Smith National Steel Corp. Trenton, MI 48183
On Tue, 26 Apr 1994, Laurie Frederick wrote:
} Hi, } We are considering the purchase of an Energy Dispersive X-ray } Analysis (EDX) system for attachment to our SEM within the next year. } Our applications are in two areas; paper making and corrosion of } materials. } } I would like to know if anyone is aware of mailing lists etc., accessable } on the Internet, which are specifically interested in EDX. I would like to } follow discussions on relative merrits of detector and window } types , vacuum requirements for windowless detectors, pulse } processor requirements, software options and in particular, read any } horror stories associated with a given manufacturer. } } If anyone has opinions on what is currently available from recent } purchase deliberations or would like to recommend a review paper on } current technology or a periodical which they go to for this sort of } information it would be greatly appreciated. } } My first priority is always dependable service, but having said that, I } will be looking for a high quality detector capable of light element } analysis, high pulse throughput, good quantitation with ease of use , } beam control and image acquisition and replay capabilities. } } Thanks for your help. } } ______________________________________________________________________ } Laurie Frederick, A.SC.T. PAPRICAN } Corrosion Control Group 3800 Wesbrook Mall } The Pulp and Paper Research Vancouver, B.C. } Institute of Canada Canada V6S 2L9 } } Email: frederick_laurie-at-vanlab.paprican.ca } Tel: 604-222-3200 Fax: 604-222-3207 }
On 27 Apr 1994 MARK-at-prl.pulmonary.ubc.ca wrote of his need for a slide with a special grid on it.
My 1992 McCrone Accessories catalog has a similar slide called an "England Finder" (p/n 313). I would call them and see what they have for you. Their phone nos. are 708-887-7100 and 800-mac-8122.
1994 PACIFIC NORTHWEST EM SOCIETY SPRING MEETING on "Evolution in Microscopy: Interfacing with Computers" Fred Hutchinson Cancer Research Center - (South Lake Union) Pelton Auditorium, Building B 1100 Fairview Ave. N. Seattle, WA 98109 Friday, May 13th
12:55 p.m. Welcoming Remarks Charles Meshul, President, PNEMS VA Medical Center, Portland, OR
1:00 p.m. "The Merging Roles of EM, Microanalysis, & Computers for Characterization of Materials." Nestor Zaluzec, Materials Science Division, Argonne National Lab
1:45 p.m. "The Evolving Role of Hardware in Image Processing: What are the Requirements?" Leonard Pagliaro, Bioengineering, Univ. of Washington
2:15 p.m. Tour of New Fred Hutchinson Facility and Coffee Break
3:15 p.m. "Analysis and Handling of Cosmic Dust." Don Brownlee, Astronomy, Univ. of Washington
4:00 p.m. "Biological Applications of Microscopic Image Analysis." Grace Bartoo, Bioengineering, Univ. of Washington
4:30 p.m. "SEM Photography: Analog/Digital; B&W/Color." David Scharf, Los Angeles, CA
5:15 p.m. Adjourn
5:30 p.m. Social -at- Benjamin's on Lake Union Sponsored by: PNEMS & Corporate Members (EMS/Diatome US, Gatan Inc., JEOL USA Inc., Link Analytical/Oxford Instruments Inc., Palmborg Associates, Philips Electron Optics)
1994 PACIFIC NORTHWEST EM SOCIETY SPRING MEETING on "Evolution in Microscopy: Interfacing with Computers" Fred Hutchinson Cancer Research Center - (South Lake Union ) Rooms B1072 and B1074, Building B 1100 Fairview Ave. N. Seattle, WA 98109 Saturday, May 14th
11:00 a.m. Adobe Photoshop Software Demonstration David Scharf , Los Angeles, CA 90039
12:00 p.m. Lunch
1:00 p.m. PNEMS Business Meeting
1:30 p.m. Video Microscopy Demonstration Gary Crawford, Mideo Systems, Huntington Beach, CA
2:30 p.m. Particle Atlas Electronic Edition CD-ROM Demonstration Steve Shaffer, MicroDataWare, Hayward, CA
3:30 p.m. Real Time Imaging and Electron Diffraction Demonstration Dave Joswiak, Astronomy, Univ. of Washington
4:30 p.m. Adjourn
* This software exchange will consist of the MSA, MAS and EMC Public Domain Library having an excess of 100 megabytes of software (Mac and PC). If you are interested, bring formatted disks to make copies of the available software.
Contact : Mike Rock -at- (206) 685-7073 or Barbara Reine -at- (206) 543-1955 for Information.
PNEMS Spring Meeting Registration Form
This meeting, (May 13 & 14) covering the topic of computers interfacing with microscopes, promises to be exciting and educational. Our speakers representing a wide range of disciplines and interests, all share an expertise in using computers in their microscopic investigation. Please note the correction of the address for the meeting (the NEW Fred Hutchinson Cancer Research Center on South Lake Union, 1100 Fairview Ave. N.) Please fill out, and return, the following form to help us plan for your attending the meeting.
Registration in advance: Students...FREE Others...$ 15.00 Registration on site: Students $ 5.00 Others $ 20.00
Name:_____________________________________
Employer: Company __________________________________________
Chris Krenn asked about experiences with intensified CCD cameras. We have a Pulnix on our high-voltage electron microscope, albeit for a different application. We use it primarily for scanning grids and low dose focussing for beam-sensitive specimens. Under conditions where the illumination is barely visible on our high-brightness screen, but no details are visible, we can clearly see all we need with the video system. We can focus to the nearest setting on the objective fine control (.99 microns per step) in a few seconds, and can focus on the vernier control (1.6 microns full scale) in a few more seconds. This system is great for our applications. There is a trade-off be- tween sensitivity and resolution--especially for the inherently low-contrast images seen at high voltage. We opted for high sensitivity, and, within that constraint, maximized the resolution. We do NOT use the system for collecting ED data, since the intense cen- tral spot would damage the intensifier, even at low-dose conditions. Further- more, it is not at all clear that the CCD gives as good quantitation and sensi- tivity as LoDose x-ray film. The first point may not be a consideration for Chris's application, but the second might. Does anyone know a system with the sensitivity to quantitate small num- bers (1 or 2) of electrons, with the ability to produce a signal proportional to electron number for the more intense reflections (i.e. no saturation)? Since each electron hitting a film grain exposes that grain, it's hard to beat that quantum efficiency. Positional accuracy is less important for us than accurate quantitation, including background subtraction.
"The preparation of samples for [electron] microscopy was still a delicate task requiring skilled human hands; the preparation of a good sample was as demanding a craft as that ever practiced by an artisan -- and took almost as long to learn." Michael Crichton The Andromeda Strain
When this quote first surfaced during lunch a few weeks ago, microscopists were asking:
"Why is specimen preparation for microscopy, whether electron or light, not simple?
"Do I have to be an artisan? After all, I've got microwaves and high tech equipment - and hey, who's got the time?
"Is electron microscopy really only science fiction?
If these and similar questions have troubled you way past midnight, then this year's Spring Symposium may be for you. The application of microwaves is revolutionizing the preparation of biological specimens and, as a bonus, is drastically reducing the time involved. New techniques for minimizing damage in the preparation of materials such as ceramics, polymers, composites and biomaterials are here. Our speakers will be telling us all about these methods.
SPECIMEN PREPARATION
SHERATON INN, MIDWAY I-94 at HAMLINE AVENUE ST. PAUL, MN THURSDAY - MAY 26, 1994
SCHEDULE OF EVENTS
8:00 - 8:30 AM Coffee and Late Registration
8:30 - 8:35 AM Welcoming and Opening Remarks. Dr. Mark Cavaleri, Research Specialist, 3M Analytical & Properties Research Labs, 3M Company, St. Paul, MN.
8:35 - 9:25 AM Preparation of Advanced Materials (ceramics, composits, biomaterials) for the Microscope Dr. Don Zipperian, Manager, Long Range Planning and Development, Buehler, LTD., Lake Bluff, IL.
9:25 - 10:15 AM Specimen Preparation of Polymers for TEM Analysis and Special Applications Ms. Jacqueline Aguilera, Senior Physicist, 3M Company, Corporate Research Analytical, St. Paul,MN.
10:15 - 10:35 AM \ Coffee Break \
0:35 - 11:25 A.M. Quantitative Imaging in the Cereal Industry: Minimizing Specimen Preparation Dr. Gary Fulcher, Professor of Food Science, University of Minnesota, St. Paul, MN
11:25 AM - 1:00 PM 5 LUNCH 6 Box lunches will be provided to participants by MMS and catered by the Sheraton Inn.
1:00 - 1:10 PM Short Business Meeting Ratification of Bylaws, Election of Officers (read Bylaws below)
1:10 - 2:00 PM Rapid Microwave Fixation of Biological Specimens for Light & Electron Microscopy Dr. Gary Login, (Part One), Departments of Pathology at the Harvard School of Dental Medicine, Harvard Medical School, and the Beth Israel Hospital, and the Charles A. Dana Research Institute, Boston,MA.
2:00 - 2:50 PM Impervious Biological Specimens: Techniques and Tricks Ms. Virginia Lindley, Dept. of Agronomy, University of Arizona; Consultant for Industrial and Federal Agencies; President of Arizona EM Society.
2:50 - 3:10 P.M. \ Coffee Break \
3:10 - 4:00 P.M. A Toolkit for Calibrating and Standardizing Microwave Fixation and Staining Dr. Gary Login, (Part Two).
------------------------------------------------------------------------------- Please make your reservation in advance, POSTMARKED NO LATER THAN MONDAY, MAY 23, if you plan to attend the Symposium. Symposium Fee: $20.00 current regular MEMS/MSOM members 93/94, $30 non-member(confers regular membership), $10.00 student members 93/94, $15.00 non-member students(confers student membership). Fill out the form near the end of this newsletter and send it in as directed(prefered) or pay at the door. -------------------------------------------------------------------------------- - REGISTRATION FOR MEMS/MSOM SPRING SYMPOSIUM, MAY 26, 1994, MIDWAY SHERATON (if your registration includes a new membership, fill out and include MMS Membership Form, below). Please postmark your reservation no later that Monday, May 23.
Name__________________________Phone____________Affiliation______________________ __ # Individual Members -at- $20.00 each = $_______. # Student Members -at- $10.00 each = $________. Total Amount Enclosed = $________. Above cost for current 93/94 MEMS/MSOM members only. Non-members and renewals please include membership dues and form(see above).
Make out your check to MMS and mail it together with this form to: Dwight Erickson, MMS Treasuresr, 3M Center, Bldg. 251-1A-03, Saint Paul, MN 55144. Late reservations may pay at the door.
------------------------------------------------------------------------------- MMS(MEMS/MSOM) Membership Form: 1993-94 All microscopists are urged to support their Society at one of the membership levels offered below. The more dues-paying members we have, the more likely we are to attract sustaining corporate member- ships which form the financial backbone of our Society. Often, supervisors will support MMS member- ships out of their project budget because they recognize that it is a very inexpensive way to maintain and increase the skills of their microscopists. If you have been a member over the years and recognize the of MMS to the community of microscopists it serves, consider upgrading your membership this year to the patron or sustaining level. Thank you. Name_______________________________ Dr____ Mr____ Ms____ Phone ( )__________ Affiliation_________________________________________Position____________________ _ Address_____________________________________________________ ZIP__________ Describe your areas of interest; state manufacturer and model of instrumentation below: Bioscience___________________________________________________________________ Materials Science ______________________________________________________________ SEM____________________ TEM______________________ X-ray_____________________ Are you an MSA Member?_______ MAS Member?_______ Other Professional groups?___________ Basic $10___ Patron $25___ Sustaining $100___ Student $5___ Due by Dec 31, '93. Make checks payable to MMS and mail to our treasurer: Dwight Erickson, MMS Treasurer, 3M Center, Bldg. 251-1A-03, Saint Paul, MN 55144.
-- Gib Ahlstrand 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
Mime-Version: 1.0 Content-Type: text/plain; charset="iso-8859-1" Content-Transfer-Encoding: quoted-printable To: Microscopy-at-anlemc.msd.anl.gov
Bill Tivol open the discusion about CCD cameras to the possible alternatives to phtographic film recording.
Did you consider the electron images plates (Fuji / Kodak / JEOL)? As far as I know (from the litterature), the DQE is about the same or better than for photographic films. The linearity is better, the dynamic range larger and they are suitable for electron diffraction. If somebody is interested I can look in my files to find more details and some references to papers (Ultramicroscopy ... a couple of years ago). Of course the complete system is a quite expensive investment(although compared to a microscope....) and seems to be available only for JEOL microscopes.
It would be nice to hear how they perform in practice from people who use them daily.
__________________________________________________________________ Philippe Buffat Ecole Polytechnique Federale de Lausanne (EPFL) Institut Interd=E9partemental de Microscopie Electronique Address: EPFL-I2M, Batiment MX-C, CH-1015 Lausanne, Switzerland Phone: +41(21)693 29 83 (Central European Time) Fax: +41(21)693 44 01 ______________________ Eudora 2.0.2 __________________________________
I also had regular problems with scratches on Dupont Cronar film. I have seen no scratches on 4489.
On Wed, 27 Apr 1994, L. D. Marks wrote:
} We currently use Agfa Scientia film for (300kV) HREM. } It is a little slower (maybe) than Kodak SO163, faster than } 4489. It has a slightly finer grain than Kodak SO163, which } helps. } Two questions: } 1) What are people using for faster film. The Agfa film } is quite old (i.e. it has been around for at least ten years), } and is there anything better at a reasonable price? } 2) We have noticed some scratches at times which seem to } be a quality control issue in the film. Is this common? } } Thanks } } Laurie Marks } Northwestern University
} From: IN%"pataky-at-bcu.ubc.ca" } Subj: Cryostat help! } } Return-path: {BIOSCI-REQUEST-at-net.bio.net} } Received: from net.bio.net by gnv.ifas.ufl.edu (PMDF V4.2-11 #3240) id } {01HBOOXBZ5DS8X4B47-at-gnv.ifas.ufl.edu} ; Wed, 27 Apr 1994 22:18:06 EST } Received: (from daemon-at-localhost) by net.bio.net (8.6.8.1/8.6.6) id TAA23358 } for cytonet-list; Wed, 27 Apr 1994 19:04:06 -0700 } Received: (from news-at-localhost) by net.bio.net (8.6.8.1/8.6.6) id TAA23346 for } cytonet-arpanet; Wed, 27 Apr 1994 19:04:05 -0700 } Date: Wed, 27 Apr 1994 23:50:46 +0000 (GMT) } From: pataky-at-bcu.ubc.ca (Dave Pataky) } Subject: Cryostat help! } To: cytonet-at-net.bio.net } Message-id: {pataky-270494154951-at-steevlab.generes.ca} } Content-transfer-encoding: 7BIT } Followup-To: bionet.cellbiol.cytonet } NNTP-Posting-Host: steevlab.generes.ca } } Anybody out there know why Tissue-Tek, presumably designed for embedding } samples for cryostat cutting, is so $##-at-***%* annoying to work with? The } problem: I'm cutting chick embryonic brain tissue, anywhere from 10-40 } microns, at -20 and no matter what I try the sections won't stop curling } up, rolling into tubes as soon as I lift the antiroll plate. I've tried } adjusting the blade angle, the "anti-roll" (that's a joke) plate, the } temperature, speed of cutting, new blade (disposable), nothing works. } Ideally I'd like to see nice flat sections that stay that way, preferably } "ribboning" off the blade so I can mount several at once. Know any tricks } or alternatives to Tissue Tek which may work better? Any ideas how to deal } with static electricity? (sometimes the sections stick to the "anti-roll" } plate). Feels like I'm battling the antichrist (and losing!), } } } -- } Dave Pataky } Dept of Zoology, UBC } pataky-at-bdc.ubc.ca } Brain can be particularly difficult. I suggest cryoprotecting in 20% sucrose with 3% PEG MW 400. Cut at -25. Don't expect to get a ribbon. We are usually satisfied with one section at a time. Rarely do we get two. I have a nice handout on cryosectioning that I picked up at a meeting. Give me your FAX # or mailing address and I will send a copy.
We were using SO163 but switched back to 4489. The SO163 seemed to have emulsion problems (much grosser defects than mere scratches, we had big chunks of emulsion fall out). We could never identify anything we were doing wrong and it seemed to be specific to certain Kodak lot numbers. We have not had problems since switching back to 4489 but sometimes need the faster film. Anyone else having problems with SO163 or should we try it again? We can't find a supplier of the Agfa film.
On Thu, 28 Apr 1994, Rodney L Kuehn wrote:
} } I also had regular problems with scratches on Dupont Cronar film. I } have seen no scratches on 4489. } } On Wed, 27 Apr 1994, L. D. Marks wrote: } } } We currently use Agfa Scientia film for (300kV) HREM. } } It is a little slower (maybe) than Kodak SO163, faster than } } 4489. It has a slightly finer grain than Kodak SO163, which } } helps. } } Two questions: } } 1) What are people using for faster film. The Agfa film } } is quite old (i.e. it has been around for at least ten years), } } and is there anything better at a reasonable price? } } 2) We have noticed some scratches at times which seem to } } be a quality control issue in the film. Is this common? } } } } Thanks } } } } Laurie Marks } } Northwestern University } } } }
Mime-Version: 1.0 Content-Type: text/plain; charset="iso-8859-1" Content-Transfer-Encoding: quoted-printable To: microscopy-at-anlemc.msd.anl.gov
Bill Tivol open the discusion about CCD cameras and the possible alternatives to phtographic film recording.
Did you consider the electron images plates (Fuji / Kodak / JEOL)? As far as I know (from the litterature), the DQE is about the same or higher than for photographic films. The linearity is better, the dynamic range larger and they are suitable for electron diffraction. If somebody is interested, I can look in my files to find the references of some (old) papers (Ultramicroscopy ... a couple of years ago). Of course the complete system is a quite expensive investment(although compared to a microscope....) and seems to be available only for JEOL microscopes.
It would be nice to hear how they perform in practice from people who use them daily. Yours
__________________________________________________________________ Philippe Buffat Ecole Polytechnique Federale de Lausanne (EPFL) Institut Interd=E9partemental de Microscopie Electronique Address: EPFL-I2M, Batiment MX-C, CH-1015 Lausanne, Switzerland Phone: +41(21)693 29 83 (Central European Time) Fax: +41(21)693 44 01 ______________________ Eudora 2.0.2 __________________________________
We have been using Kodak SO163 without difficulty for years and we now use it in our JEOL JEM-4000EXII. We started using it because it is faster than 4489 and it can be pushed. The slightly larger grain does not seem to be a problem for our HREM users, but most do want increased film speed.
Russell Cook Electron Microscopy Center for Materials Research Argonne National Laboratory Argonne, IL 60439
Can someone offer suggestions for photographing Bacillus subtilis at the optical microscope level? These bacteria are small, motile and contain refractile bodies. In a fluid media there is a lot of Brownian motion and the refractile bodies create diffraction rings. We were thinking of using methyl cellulose. Can anyone suggest the proper viscosity to use? Does anyone know of a book or book chapter describing photographing bacteria?
I am just getting started trying to do low-dose HREM at high (1 MV) voltage. I have been told that SO163 developed in undiluted D-19 for 12 min is the best way to go, and I plan to try it as soon as the film arrives. Mean- while, I have been working at high mag (200kx-400kx) with Dupont LoDose. It is at least an order of magnitude more sensitive than SO163, but the grain size is also an order of magnitude larger (makes sense, equal sensitivity per grain and all that). Other drawbacks are the blue backing for LoDose and the tendency to fog and to show arrowhead-like marks from static electric discharges when very dry films are separated. Working in total darkness is also required with Lo- Dose. Murray King, at our lab, did a systematic study of the developing and fixing conditions for 4489 and LoDose to produce the best combination of sens- itivity and low fog. As a result we use 4 min in D-19 (4489) or GBX (LoDose) and 5 min fix. I'd be interested in anyone else's results with SO163. Although tedious, I believe such systematic studies are very valuable. If this particular wheel has not already been invented, I'll probably undertake the study in my copious free time (to quote from T. Lehrer).
Chris Krenn asks about using SIT cameras for detecting backscattered electron diffraction patterns.
As I understand this problem, you may not need the full sensitivity of the SIT (10^-4). A good CCD camera with exposure control may do the trick. A sensitive camera would have about .5 lux sensitivity, or less, with a nod given to the noise floor, at 1/60 sec exposure. Integrating on the chip for under a second would give around 10^-2 lux, with more available at longer times. This may be enough.
This camera would cost, maybe, $1400, or around 8% of your SIT.
My company makes a device called the OMNEX which can control these cameras, as well as do real time averaging, memory functions, measurements, digital contrast control, psuedocolor, zoom, frame storage, and lots of other things. We have a lot of people using it for applications where a little longer exposure time is all that's needed. (We also have people using it with SIT cameras for all of its other functions.)
Good luck. Send me a note if you would like any more info.
Bill Tivol posted a message regarding films for low-dose HREM. Perhaps I can suggest to use CCD camera (if one can afford it) to do a much better job. I have been doing low-dose HREM for several years now, and after taking the pains and frustrations with films, we started using slow-scan CCd camera 3 years ago. We are very pleased with the performence of the camera, and able to record low-dose HREM images with much confidence. I think CCD cameras offers the following advantages over conventional films: (1) Digital recording, i.e. on-line evaluation of image quality and fine tuning microscope operating conditions; (2) Very high sensitivity (low-dose). I am going to give a talk on low-dose HREm in the upcoming MSA meeting. If anyone is interested, please send me an email.
Ming Pan CSSS, Arizona State University panm-at-csss.la.asu.edu
Laurie asked for help on EDX selection, in this case windows:
Disclaimer: MOXTEK makes beryllium windows and ultra-thin windows for Si(Li) detectors, and supplies about half of the Si(Li) windows used world wide. I honestly tried to be objective in the following, but needed to warn you! I would be very interested in any comments, anecdotes, etc. from the microscope community on this subject.
I just wrote a chapter on thin windows for light element analysis for the book that Dave Williams and Dale Newbury are editing for the MAS. The book is entitled "X-ray spectrometry in electron beam instruments." It should be out this fall. A few items from my paper may be helpful:
If you are interested in light element analysis you should buy the spectrometer as a whole instrument for light element analysis. There is a lot more to light element sensitivity than the window (detector, FET preamp, processing electronics, software). It is possible to buy the best window, but still not get the best detection limits.
Windowless detectors have generally not lived up to their potential because of icing problems. A layer of ice can absorb as much as a thin window. If your microscope is very clean and scrupulously maintained vacuum-wise, however, this may be a good option. (This subject needs more text to treat fairly: manufacturers have done a good job to provide de-icing cycles, etc. to solve these problems.)
It is essential to maintain a steady regimen of standards testing to know the condition of the detector with all Si(Li) detectors. Two major things that can go wrong are detector icing and loss of vacuum (which will warm the detector, causing an increase of noise.)
Sometimes, to obtain better sensitivity at Be and B, a thin window is used that does not have an aluminum light blocking layer. Consider whether you need to block light from the instrument, room, or sample before selecting one of these. Even with aluminum coatings ultra-thin windows leak more light than beryllium windows do.
Since you are putting the spectrometer on an existing SEM it is a good idea to discuss your selection with both the electron microscope company and the spectrometer company. The biggest failure mechanism of thin windows is impact of particles on the window during venting. Some microscope models never have a problem, and some have a big problem. There are 'fixes' for the venting systems of problem microscopes.
The traditional reason for using something like dry nitrogen is degassing from the walls of a vacuum system, which is the major vacuum limitation (plus to some extent) of an unbaked system. If one uses dry nitrogen, then the initial chemisorbed layers tend to be quite nitrogen rich; nitrogen desorbs easily. If instead one uses air, then water and hydrocarbons (car exhaust fumes, people's breath etc) chemisorb on the walls and only very slowly leave. I must admit that with a modern microscope there is probably not that much of an effect since pumps have come a long way in pumping speed/$. However, if you are really concerned with UHV systems or are fighting contamination problems nitrogen (with very low hydrocarbon content) is probably going to help. I am not sure that the water level will be very relevant for most people.
Addendum: My mailer ate part of my last message. Leaks are also important, but one should worry more about backstreaming from roughing pumps in my experience.
Subject: Time:4:10 PM OFFICE MEMO Re: Backfill gases Date:4/29/94 This is a topic that I have discussed in some detail in a book on "Vacuum Methods in Electron Microscopy" that will be pub- lished early in May as the latest volume in Audrey Glauert's series PRACTICAL METHODS IN ELECTRON MICROSCOPY. Briefly, the purpose of backfilling with a dry gas is to reduce the amount of water vapor that is adsorbed onto the surfaces inside the vacuum system when the system is let up to atmospheric pressure. In even a moderately humid environment, or under conditions of prolonged exposure to the atmosphere, a surface can become covered with up to a hundred molecular layers of adsorbed water. Because water molecules are highly polar they adsorb to most surfaces quite tenaciously, and then desorb only very slowly when the system is pumped down again. This greatly prolongs the time required to pump down to an operating vacuum. Furthermore, unless a system can be baked out at temperatures above 200#161#C, the desorption of water also makes it very difficult to reach pressures much below the upper end of the 10-7 Torr range. You are correct, however, in concluding that there is little benefit to admitting a dry gas into the photographic chamber of an electron microscope, simply because the water evolved by the film will quickly coat the surfaces with water anyway. The same applies to bell jars, the specimen chambers of SEMs, electron guns, and other systems which are left standing open to the atmosphere for long periods of time. However, in situations where only a small hole will be opened in a vacuum system, such as when an aperture manipulator is being serviced, it is beneficial to use the dry gas, and to plug the opening loosely with aluminum foil and to maintain a slow flow of dry gas through the system while it is at atmospheric pressure. The primary concern in selecting the gas to use is that it be free of both water vapor and oil vapor. Gas pumped with an oil-sealed compressor will quickly lead to a very high rate of hydrocarbon contamination on the specimen. Other than that, nearly any dry gas will work, even dry air. Dry nitrogen is so commonly used because it is so readily available. As described in my book, it is even possible to capture the nitrogen that boils off from a Dewar flask in a plastic balloon and to use it for this purpose.
Comments: Converted from PROFS to RFC822 format by PUMP V2.2X
Thanx for the info about the image plate and CCD talks at MSA 94. I'll be at both. In both cases, the delay in readout, etc. is nothing compared to the time it takes to scan a negative with 10*10 micron pixels and to prepare the file to be small enough so that our computer doesn't choke on it. For HREM, digitization via CCD is really the way to go, and we intend to go that way if the next renewal of our grant allows us to purchase the equipment. For quan- titation of ED patterns, I don't know whether the CCD might not be overloaded at the center spot--our intensified CCD can be damaged by trying this. From the viewpoint of "making every electron count", can either method sense a single electron--actually, if each electron produces n photons, where n} } 1, this sensitivity is not as tough as it might first appear. There might be an interesting idea in trying a thicker phosphor or YAG for ED and a thinner (thus better resolution) one for HREM. Has anyone investigated this yet? In parti- cular, for our 1.2 MV electrons this makes a lot of sense to me.
Re the discussion of water-cooled xma detectors, does anyone make or use a de- tector cooled by a Peltier device? I used such a detector many years ago for proton detection. There, the detector was a silicon junction detector and was cooled in order to reduce noise; whereas, the LN cooling of Si(Li) detectors is essential to preserve the drift profile.
Alex King asked about the reasons for the use of a dry gas for airing the vac- uum in a microscope. Others have answered the why. The grade we use is the "HP nitrogen"; the UHP seems to be no better and much more $.
The detector in an EDX system is cooled to liquid nitrogen temperatures to lower the noise due to thermally generated carriers. A second reason the detector is cooled is to keep the lithium from drifting around, but modern systems can be warmed if the detector is biased.
The systems that seem to be water cooled are actually cooled with thermoelectric coolers to cryogenic temperatures. The water is used to carry away the heat from the TE coolers, which is considerable. The TE coolers cannot get the detector to 77K, which is why these systems are noisier than the LN cooled systems.
regards Mark W. Lund, PhD Director MOXTEK, Inc. Orem UT
On 27 Apr 1994 MARK-at-prl.pulmonary.ubc.ca wrote of his need for a slide with a special grid on it. I was unable to reach him with e-mail.
My 1992 McCrone Accessories catalog has a similar slide called an "England Finder" (p/n 313). I would call them and see what they have for you. Their phone nos. are 708-887-7100 and 800-mac-8122.
We use another method to provide dry nitrogen for venting, valves etc.. Rather than use bottled gas, we take the gas from our liquid nitrogen vessel. It is dried and as yet (touch wood) we have had no problems. This is after approximately 18 months operation.
The vessel is large, 2000 litre capacity but the gas is used for a TEM, SEM, quadrupole mass. spec. and a few other instruments. It is also used to vent ALL vacuum systems in the E.M. lab.
It sure beats having to change gas bottles!!
##################################################################### ********************** * Between the idea * 0------* And the reality * } ---|--- { * Between the motion * | * And the act * / \ * Falls the Shadow * _/ \_ * T.S. Eliot * ********************** Colin Veitch Tel + 61 (0)52 47 2611 CSIRO Division of Wool Technology Tel + 61 (0)52 47 2891 (dir.) P.O. Box 21 Fax + 61 (0)52 47 2657 BELMONT Vic 3216 Australia
To biological light microscopists, I want to make hydrophobic rings on glass microscope slides so that tissue-processing and immunocytochemistry can be carried out on tissue on the slide. To date, I have been smearing a thin ring of silicone (auto and caravan sealant!) and allowing it do dry. The difficulty with this is that the silicone has to be shaved off with a razor blade before the tissue is cover-slipped. I have heard of a product called a PAP pen which does the same thing, with the advantage that the hydrophobic layer is very thin. I found one such product made by "Agar" Essex, UK, distributed by "Alltech" (Australia) but the asking price seems outrageous, approx $Aus120 per pen (approx $US80). 1. Is there a similar product available from other companies (addresses, fax. nos appreciated)? 2. Is there a "home-brew" alternative, perhaps using liquid silicone?
Thanks for any advice, Dave Merritt Zoology University of New England Armidale NSW Australia dmerritt-at-metz.une.edu.au
I used to use a Sorval MT2-B. The company that serviced it for me was "RMC" in Arizona. Their phone number is 602-889-7900. They have service people all over the US. You may want to try them.
I've been charged with purchasing a high resolution coating system to use with both scanning probe and electron microscopes. My lab is primarily a scanning force microscope lab where we look at proteins and polysaccharides on polymers and cell membranes. We are interested in starting a substantial correlative microscopy effort using field emission SEM's on the same samples that we've previously probed with AFM or STM. The only coating systems currently available to us are mechanically pumped DC sputtering units. I have ~$25k to spend and have spent some time looking at the options. It looks like a turbo pumped sputtering system with a rotary tilt stage would best suit our needs. However, I'm in the dark concerning what parameters to assess in determining an optimal system. I know that grain size varies with the sputtered material. Does it also vary between different sputtering systems? Is a thermal evaporation source something to consider? Is it important to spend the money on an oiless backing pump for the turbo? What questions am I not asking that I should be asking?
To make hydrophobic rings on glass microscope slides an old homebrew method is to dilute a resin mountant such as DePex or Permount or Canada Balsam in the appropriate solvent and to use this as ringing agent. Try diluting the mountant 10 to 50 times with the recommended solvent (usually xylene or toluene) and apply with the sharpened end of an applicator stick. The advantage is that you do not have to remove the stuff before mounting under a coverslip. It is also a useful method to keep serial monitor sections in the correct order, each on its own separate little water droplet inside its own hydrophobic ring.
Jan Coetzee Unit for Electron Microscopy Tel:+27-12-420-2075 University of Pretoria Fax:+27-12-342-1738 Pretoria Internet:janc-at-ccnet.up.ac.za 0002 South Africa
We are using fluorescent dyes on bleached wood pulp fibres (ie cellulose) for confocal microscopy. We are having problems with "bleeding" or leaching of dye. Are there any fluorescent dyes which are reactive with cellulose or can be fixed similar to textile dyes (RIT, etc.) so that they will not leach out in aqeous environments? Thanks in advance.
James Drummond Pulp and Paper Research Institute of Canada Vancouver, B.C. Canada
We are using fluorescent dyes on bleached wood pulp fibres (ie cellulose) for confocal microscopy. We are having problems with "bleeding" or leaching of dye. Are there any fluorescent dyes that bind to cellulose or can be fixed similar to textile dyes (RIT, etc.) so that they will not leach out in aqueous environments? Thanks in advance.
James Drummond Pulp and Paper Research Institute of Canada Vancouver, B.C. Canada
} There is a potential alternative for bottled N2. There are driers for } compressed air that dry it to a lower dew point than lab grade N2. In } theory bleeding some of this into the tank instead of N2 from a } cylinder should work.
I haven't tried this, but you still would probably have the problem of all the hydrocarbons from a standard air compressor... How do these driers work? If they use some kind of cold trap, you might be okay. Liquid nitrogen will freeze both water and hydrocarbons out of an atmosphere.
Chris Krenn Graduate Student UC Berkeley Dept. of Materials Science
At the risk of swimming upstream (and against the apparent popular opinion), I have used PAP pens for slide mounted tissue processing, and if that is your method of choice, I think the PAP pen works just fine. I say if that is your choice only because i much prefer processing immunocytochemical material as free-floating sections rather than slide-mounted - better penetration, more uniform staining, and (extremely important) better rinsing (at least in my experience). The PAP pen, as best as I can smell, is a mixture of bees wax dissolved in xylene (or some similar concoction). It goes on easily if you have a dry area around your tissue, and it comes clean in a xylene rinse before mounting. It requires a little practice to get the right pressure so you don't get too much liquid out of the pen, but it is not brain surgery!
The PAP pen is available for about $30 US from RPI:
Research Products International 410 North Business Center Drive Mount Prospect, IL 60056
Ph: 1-800-323-9814
Let me know if you have any further questions. Cheers,
David Morilak Dept Pharmacology Univ Texas Health Science Center San Antonio, TX 78284
Some of the problems can be due to using a disposable blade. We cut routinely serial cryosections of young chick brains cyroprotected in 30% sucrose and snap frozen in dry-ice or nitrogen chilled heptane and mounted on the chuck with OCT. Fixation is with just about every imaginable fix possible. Crank the temperature down, I like -25 to -35, use a real blade and keep it clean, especially the back of the blade. Sometimes I swing the anti-roll plate out of the way and use a #2 paintbrush (keep it inside the cryostat) to prevent curl - touch it to the base of the section as it gbegins to come off the blade and coordinate your arm so that you can move the brush in rythmn with the section as it moves with the cutting stroke. I've tried disposable blades for paraffin, to cut serial paraffin sections, without success. But the standard blades will allow me to cut ribbons as long as my arm can stretch holding the free end of the ribbon.
On Thu, 28 Apr 1994, Greg Erdos ICBR EM Core Lab Univers wrote:
} From: TOWER::GWERDOS "Greg Erdos ICBR EM Core Lab Univers" } To: IN%"pataky-at-bcu.ubc.ca" } CC: GWERDOS } Subj: Re: Cryostat help! } } } From: IN%"pataky-at-bcu.ubc.ca" } } Subj: Cryostat help! } } } } Return-path: {BIOSCI-REQUEST-at-net.bio.net} } } Received: from net.bio.net by gnv.ifas.ufl.edu (PMDF V4.2-11 #3240) id } } {01HBOOXBZ5DS8X4B47-at-gnv.ifas.ufl.edu} ; Wed, 27 Apr 1994 22:18:06 EST } } Received: (from daemon-at-localhost) by net.bio.net (8.6.8.1/8.6.6) id TAA23358 } } for cytonet-list; Wed, 27 Apr 1994 19:04:06 -0700 } } Received: (from news-at-localhost) by net.bio.net (8.6.8.1/8.6.6) id TAA23346 for } } cytonet-arpanet; Wed, 27 Apr 1994 19:04:05 -0700 } } Date: Wed, 27 Apr 1994 23:50:46 +0000 (GMT) } } From: pataky-at-bcu.ubc.ca (Dave Pataky) } } Subject: Cryostat help! } } To: cytonet-at-net.bio.net } } Message-id: {pataky-270494154951-at-steevlab.generes.ca} } } Content-transfer-encoding: 7BIT } } Followup-To: bionet.cellbiol.cytonet } } NNTP-Posting-Host: steevlab.generes.ca } } } } Anybody out there know why Tissue-Tek, presumably designed for embedding } } samples for cryostat cutting, is so $##-at-***%* annoying to work with? The } } problem: I'm cutting chick embryonic brain tissue, anywhere from 10-40 } } microns, at -20 and no matter what I try the sections won't stop curling } } up, rolling into tubes as soon as I lift the antiroll plate. I've tried } } adjusting the blade angle, the "anti-roll" (that's a joke) plate, the } } temperature, speed of cutting, new blade (disposable), nothing works. } } Ideally I'd like to see nice flat sections that stay that way, preferably } } "ribboning" off the blade so I can mount several at once. Know any tricks } } or alternatives to Tissue Tek which may work better? Any ideas how to deal } } with static electricity? (sometimes the sections stick to the "anti-roll" } } plate). Feels like I'm battling the antichrist (and losing!), } } } } } } -- } } Dave Pataky } } Dept of Zoology, UBC } } pataky-at-bdc.ubc.ca } } } Brain can be particularly difficult. I suggest cryoprotecting in 20% sucrose } with 3% PEG MW 400. Cut at -25. Don't expect to get a ribbon. We are usually } satisfied with one section at a time. Rarely do we get two. I have a nice } handout on cryosectioning that I picked up at a meeting. Give me your FAX # } or mailing address and I will send a copy. } } ********************************************************** } * Greg Erdos ** * } * Director, ICBR EMCL ** Phone 904-392-1295 * } * 218 Carr Hall ** FAX 904-392-8598 * } * University of Florida ** gwerdos-at-gnv.ifas.ufl.edu * } * Gainesville, FL 32611 ** * } ********************************************************** } ********************************************************** } * Greg Erdos ** * } * Director, ICBR EMCL ** Phone 904-392-1295 * } * 218 Carr Hall ** FAX 904-392-8598 * } * University of Florida ** gwerdos-at-gnv.ifas.ufl.edu * } * Gainesville, FL 32611 ** * } ********************************************************** }
FOR THOSE OF YOU INVOLVED IN EDITING JOURNALS, WRITING BOOKS, ETC. IN THE FIELD, AND ANYONE ELSE WITH AN OPINION, WHAT DO YOU THINK ARE THE MOST ACCEPTABLE ACRONYMS TODAY FOR: HIGH RESOLUTION (TRANSMISSION) ELECTRON MICROSCOPY " " (SCANNING) " " LOW VOLTAGE, FIELD EMISSION, SEM HIGH PRESSURE OR ENVIRONMENTAL SEM
WOULD APPRECIATE SHORT RESPONSES WITH SUGGESTED ACRONYMS, TKS, LINDA SAWYER
We have a JEOL JSM-35C SEM in outstanding condition for sale. It was purchased in 1980 and was on service contract until April 21, 1994. It comes with a Tracor Northern (Noran) EDS detector and TN-2000 analyzer. Asking $8,000.00. Contact Dr. Stanley L. Flegler Center for Electron Optics Michigan State University Flegler-at-pilot.msu.edu
FYI, using FETCH, I reached your ftp volume at pub/ncsu/jruss, not pub/jruss as indicated in your note to the NIH Image list. Would this be a better place to exchange things rather than mailing?
Bill ========================== Bill Heeschen/Analyt. Sci.- Mat. Char. ----- 1897-F Building / The Dow Chemical Co. --- --- Midland, MI 48667 U.S.A. --- D O W --- phone: (517)636-4005 fax: (517)636-5453 --- --- Email: waheeschen-at-dow.com ----- ========================== R
Microscopy list: My apologies for that last note. It was intended for John Russ in response to one of his helpful explanations on the NIH Image mail list.
Again, my apologies.
Bill ========================== Bill Heeschen/Analyt. Sci.- Mat. Char. ----- 1897-F Building / The Dow Chemical Co. --- --- Midland, MI 48667 U.S.A. --- D O W --- phone: (517)636-4005 fax: (517)636-5453 --- --- Email: waheeschen-at-dow.com ----- ========================== R
If there are any groups in the preperation of pulp that bind feulgen, the material will give a brilliant red signal appropriate for most confocal instruments. It is very "fast" if it binds.
Bill Tivol asked: Does anyone know a system with the sensitivity to quantitate small num- bers (1 or 2) of electrons, with the ability to produce a signal proportional to electron number for the more intense reflections (i.e. no saturation)? Since each electron hitting a film grain exposes that grain, it's hard to beat that quantum efficiency. Positional accuracy is less important for us than accurate quantitation, including background subtraction.
I don't have any inherent bias towards Hammamatsu; it is merely the only vendor whose literature I've gotten so far, and whose name I was familiar with because of their image processing boxes. They have a series of CCD based photon counters which also have a wide dynamic range (10^6)
Chris Krenn Graduate Student UC Berkeley Dept. of Materials Science
Thanks for your reply. Word of your good results came through the grapevine, and was one of the reasons we are looking at CCD instead of SIT. Hopefully we will have a somewhat functional system by the end of the year.
Received: from MAILQUEUE by MICROBIO1 (Mercury 1.11); Tue, 3 May 94 11:36:44 EST Return-path: {Stanley_Hayes-at-rml.niaid.pc.niaid.nih.gov} Received: from emoryu1.cc.emory.edu by transporter.microbio.emory.edu (Mercury 1.11); Tue, 3 May 94 11:36:37 EST Received: from anlemc.msd.anl.gov by emoryu1.cc.emory.edu (5.65/Emory_cc.3.4.21) via SMTP id AA14758 ; Tue, 3 May 94 12:34:03 -0400 Return-Path: Stanley_Hayes-at-rml.niaid.pc.niaid.nih.gov
Dear Alfred,
We have a Pall air dryer, not to air the HVEM, but to pump dry air through our high-voltage tanks in the event that they have been opened. We find that using the dried air makes a big difference in residual water in the tanks after the air is pumped out and replaced by the SF6 insulating gas which is the normal fill gas for the tanks. These tanks are 2-3 meters in diameter and about 4 meters tall with a connecting piece about 1 meter**3. The air dryer has a pre-filter to remove oil, the dessicant, an oil vapor filter, and a particle afterfilter. The specified dewpoint reached is -60o F or below. It's also noisy enough that I'd hate to have to turn it on each time I wanted to air the microscope, but it could be mounted in a sound-absorbant cabinet.
Message-Id: {199405040807010548-at-qmgate.secrc.abb.se} X-Mailer: InterCon Dispatcher/SMTP for QuickMail X-Priority: 4
Looking for someone that has been working with oxidation, hydriding and wear properties of surface coated or surface modified zirconium base alloys (like Zircaloy 2 or 4). The environment is water or steam at ]300!C. I am interested in all topics related to the subject; coating or surface modification methods ( how to get a coating without defects going through the coating?), corrosion tests, investigation of tested samples (LOM, SEM, TEM, SAM, XRD, electrochemical impedance spectroscopy etc.).
Bengt Stridh ABB Corporate Research S-721 78 Vasteras Sweden E-mail: best-at-secrc.abb.se Fax: +46-21-13 41 00 Phone: +46-21-32 30 67
================================================ An Update Message from the Microscopy Listserver ================================================
G'day All Microscopy Listserver subscribers....
In sorting out a mailserver problem, today, I just noticed that about 2 weeks ago we received our 1000th subscription request. As in previous milestones, I hearby offer to by a beer (or any other appropriate concoction) for
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at the next microscopy meeting that we both attend. I'll definitely be at the MSA New Orleans meeting so Mikey whoever you are, make sure you look me up if your there. I hear tell there may be a pub/bar or two in town. Hmm.. at this rate (1000+ in 6 months of operation), I may be buying more than 1 or 2 beers in New Orleans, hope I don't exceed my limit being the quiet, unassuming person that I am. ;-)
We now have subscribers on every continent except Antarctica anyone know a frozen microscopist/microanalyst down there that's on Email??
As promised earlier in the year a new computer system has arrived, and I'm slowing getting around to shifting software to that machine. I'm having some trouble linking the MSA BBS into the new machine so things are taking longer than expected (as usual). The transition should be seemless at your end as there will be no changes in addresses or functionality just new hardware.
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FOR THOSE OF YOU INVOLVED IN EDITING JOURNALS, WRITING BOOKS, ETC. IN THE FIELD, AND ANYONE ELSE WITH AN OPINION, WHAT DO YOU THINK ARE THE MOST ACCEPTABLE ACRONYMS TODAY FOR: HIGH RESOLUTION (TRANSMISSION) ELECTRON MICROSCOPY " " (SCANNING) " " LOW VOLTAGE, FIELD EMISSION, SEM HIGH PRESSURE OR ENVIRONMENTAL SEM
WOULD APPRECIATE SHORT RESPONSES WITH SUGGESTED ACRONYMS, TKS, LINDA SAWY
Does anyone know of a low to moderately priced sharpener for carbon coater rods? The motorized ones I've seen in microscopy supply houses are going for about $1000 which seems a bit over-priced to me. I saw one once that was based on an ordinary manual type pencil sharpener, have not been able to find one. Any help on this subject would be greatly appreciated.
********************************************************************** * Glenn Poirier glenn_p-at-geosci.lan.mcgill.ca * * Electron Microprobe Lab Phone: (514) 398 6774 * * Earth and Planetary Sciences Fax: (514) 398 4680 * * McGill University THERE ARE THREE SIDES TO EVERY STORY; * * Montreal, Quebec YOUR SIDE MY SIDE AND THE TRUTH * **********************************************************************
The X-ray lab at the US Bureau of Mines has a carbon coater rod sharpener that is like a pencil sharpener. Where they aquired it i am not sure. You should contact Gary Van Landyut at PO Box 280, Rolla, Mo 65401 or call 314-364-3169. C
This may sound a little crazy but... Try a theatrical lighting company-one that specializes in high intensity spotlights. these apparently use carbon rods which are sharpened in a similar manner.
Mark Elliott UBC-Pulmonary Research Laboratory, St. Paul's Hospital, Vancouver, BC
I'm interested in any recommendations for cooled CCD cameras and associated MAC software for light-level immunofluorescent studies. We currently use a Hamamatsu SIT camera and NIH Image. The sensitivity is OK for our studies, but I'd like better resolution. Our cells are very small and we want to image several monoclonals against antigens in the same cell. What are the advantages of the various CCD brands? Has anyone used IPLabs software with a Power Mac? Will we see a big improvement in resolution over the SIT camera (an NTSC-based video camera) we now use? Thanks for any replies.
Does anybody have a proven method (recipe and conditions) for the electropolishing of Ta single crystal samples? Thanks. Jan Coetzee Unit for Electron Microscopy Tel:+27-12-420-2075 University of Pretoria Fax:+27-12-342-1738 Pretoria Internet:janc-at-ccnet.up.ac.za 0002 South Africa
Date 5/6/94 Subject Carbon Rod Sharpeners From Chris Bowser To Magnavox Internet
Subject: Time:7:03 AM OFFICE MEMO Carbon Rod Sharpeners Date:5/6/94 smtp%"microscopy-at-anlemc.msd.anl.gov"
I have a carbon rod sharpener that I purchased about 10 yr ago from the Ernest F Fullam company. You might try them. Also the Ted Pella company has a small hand sharpener for about $40.00.
We are planning on using freeze-substitution as our standard protocol for specimen preparation for thin sections. I would like to get an idea of how many subscribers to this newsgroup use freeze-substitution and what general comments you may have. I have several questions, some of which cannot be answered by reading published methods. We will be freezing our specimens (bacteria) in liquid nitrogen cooled liquid propane. Should a special grade of propane be used for this? What safety precautions should be used with liquid propane? What types of substitution media are preferred? I have seen that both methanol and acetone are commonly used as solvents. Most substitution media contain glutaraldehyde, osmium tetroxide and uranyl acetate. Are there any tricks to making this mixture up? Once the substitution medium has been made up, should it be stored frozen in liquid nitrogen? Thanks in advance for any comments.
J. W. Austin Microbiology Research Division Food Directorate Health Protection Branch Health Canada Ottawa, Ont.
The Microscopy Listserver has been down due to hardware problems for the last 2-3 days. Things should be back to normal shortly. Please report any major problems to: Zaluzec-at-anlemc.msd.anl.gov
Sometimes ago I posted a questions about difference between digital and other type confocal. I got an address for a server group. I tried subcribing, but could not. Does anyone has that server address, and can someone give me information about confocals.
S. D. Walck's contribution to the backfill question had a lot of good things to say. I'd like to add two cents worth: Using teflon tubing, available, e.g. from Cole-Parmer works better than tygon in LN2.
i'm looking for a U.S. distributor, if there is one, for vertical slide staining dishes and racks. like standard dishes + racks, but the slides go in vertically and use less solution, and can hold more slides (25x75)than a coplin jar - 20 or 30 i think. they're evidently standard in Japan, and we have a quote from a Matsunami Trading Co. in Osaka for them at $10.70 for racks and $11.36 for dishes, which is reasonable, but Air Parcel Post at 2/3 of the list price isn't. so i was wondering if anyone might know of a U.S. source for these things. haven't come across any in the obvious places.
------------------------------------------------------------------ |Glenn Holm Internet:karuzis-at-wccf.mit.edu| |M.I.T Dept. of Brain + Cog. Sci. Bitnet:karuzis-at-mitwccf | |Cambridge, MA 02139 "Real Neuroscientists don't do gels!" | ------------------------------------------------------------------
Try Fisher Scientific - the staining sets are plastic, hold from 100-250 ml depending on how much of the slide you need immersed, and the racks hold 25 slides. The only problem I have found using these setups is that evaporation of solution is serious if you leave them sit. I keep my solutions in bottles and pour them back when I'm finished staining (the ones that aren't made up fresh of course). The sets are made by Tissue-Tek.
Cheers
David Morilak Dept Pharmacology UT Health Science Center San Antonio morilak-at-uthscsa
I was wondering if anyone could recommend a couple of good books on TEM and SEM of plant materials. I've always worked with animal and human tissue, and microorganisms so all of my books are related to these topics.
Thanks,
Phil Rutledge p.s. If you know the publisher and ISBN that would help also.
Message-Id: {MAILQUEUE-101.940511084750.751-at-prl.pulmonary.ubc.ca} To: microscopy-at-anlemc.msd.anl.gov
Does anyone know of an easy way to fix a scratch on a 40X dry objective lens for an Olympus scope, other than returning it to Olympus??? Someone here scratched their's and are on limited budget and need it fast. Any suggestion greatly appreciated.
Thanks, Mark Elliott UBC-Pulmonary Research Lab, Vancouver
Message-Id: {MAILQUEUE-101.940511095225.756-at-prl.pulmonary.ubc.ca} To: MICROSCOPY-at-anlemc.msd.anl.gov
a FEW SUGGESTIONS; Introduction to Biological Electron Microscopy: Theory and Techniques, By Clinton J. Dawes Ladd Research Industries, Inc, Burlington Vermont, Library of Congress # 86-082930-not sure of ISBN #
Also Books by Robards, not sure of title-if find will let you know. Can also check for books by Hyatt, not sure which one but one has plant material in it.
Depends on what type of plant material you are dealing with-Fungi, marine algae or vascular plants-they are all different and their methods of preparation are different.
We have been using Denka LaB6 {100} 60 deg filaments in our JEOL 4000 with less than great results. Brightness is poor and after a week of use the center of the filament is small and dim and most of the illumination is from the lobes. Has anyone tried the LaB6 filaments from Kimble Physics or FEI in a JEOL 4000? Do they seem better or worse? Do they last?
Thanks Roseann Csencsits Electron Microscopy Center Argonne National Laboratory Argonne, IL
If anyone has a Noran 5500 no longer in service with good display boards and wants to make a deal. Please contact me directly by e-mail or voice.
*************************************** David Garrett "DGARRETT-at-GAB.UNT.EDU" University of North Texas Dept. Biological Sciences (817)565-3964 Fax (817)565-4136 ***************************************
} We have been using Denka LaB6 {100} 60 deg filaments in our JEOL 4000 } with less than great results. Brightness is poor and after a week of } use the center of the filament is small and dim and most of the } illumination is from the lobes. Has anyone tried the LaB6 filaments } from Kimble Physics or FEI in a JEOL 4000? Do they seem better or } worse? Do they last? } } Thanks } Roseann Csencsits } Electron Microscopy Center } Argonne National Laboratory } Argonne, IL
We were trying the Kimball filaments when I first got here two years ago. After blowing a few, about which they were VERY understanding and generous, we decided to go back to tungsten, until everyone could sit down and figure out what was happening.
Peter Sewell, who makes the filaments, has been very helpful and accessible. At the time, he told me that they were doing some more testing on why they were getting runaway heating in JEOL scopes, but not Philips. It has to do with the control circuitry of the gun and what is being regulated. (I'm neither a physics nor an electronics whiz, so please don't ask for more specifics from me!)
The bottom line is that conditioning of the filament to be used in a JEOL, as well as conditioning of the gun, is essential if you're going to use their filaments. We have a JEOL 2000 EXII and have adopted their procedure at installation of tungsten filaments. We haven't gone back to trying the Kimball ones yet.
Filament conditioning: after installation, set gun bias to zero, turn up filament heating SLOWLY, when the vacuum starts to degrade stop, when it stabilizes continue turning up till you get to near your normal operating conditions. Only after heating the filament and getting beyond the outgassing should you turn up the bias to get emission. After conditioning this way, you shouldn't get any more outgassing. From what I can tell, you should be able to do this at any accelerating voltage. This conditioning has taken me one and a half hours. Be patient. I'm expecting very long filament life after this treatment.
I'd be interested in others experiences with this kind of procedure.
John chandler-at-lamar.ColoState.EDU Fort Collins CO
} From: IN%"jgoodhouse-at-molecular.Princeton.EDU" "Goodhouse, Joseph" } Subj: Silver enhancement, ultra structure } } Return-path: {jgoodhouse-at-molecular.Princeton.EDU} } Received: from anlemc.msd.anl.gov by gnv.ifas.ufl.edu (PMDF V4.2-11 #3240) id } {01HC7XIRCF4G8X5IJZ-at-gnv.ifas.ufl.edu} ; Wed, 11 May 1994 16:49:27 EST } Date: Wed, 11 May 1994 09:48:00 -0400 (EDT) } From: "Goodhouse, Joseph" {jgoodhouse-at-molecular.Princeton.EDU} } Subject: Silver enhancement, ultra structure } To: Micrscopy {Microscopy-at-anlemc.msd.anl.gov} } Message-id: {2DD0E2AB-at-molecular.princeton.edu} } X-Mailer: Microsoft Mail V3.0 } Content-transfer-encoding: 7BIT } Encoding: 10 TEXT } } } I am trying to do silver enhancent on 3nm gold probes in Drosophila embryos } for ultra structure morpholgy. The problem I am experiencing is that the } reaction is occurring too rapidly and is blowing the embryos apart. Can } some one direct me to a few good references on this technique for cells and } tissues, or provide me with a working protocol. Much appreciation } Joe Goodhouse } Dept. of Molec. Bio. } Princeton University } jgoodhouse-at-molecular.princeton.edu #################%%%%%%%%%%%%%%%%%%%%%%%%%%%##################%%%%%%%%%%%%%% If you are using one of the daylight kits like the one from BioCell you might try diluting the reagents with water to slow the reaction. Their rep. told me this at a meeting one time. Maybe lowering the temp would work too ********************************************************** * Greg Erdos ** * * Director, ICBR EMCL ** Phone 904-392-1295 * * 218 Carr Hall ** FAX 904-392-8598 * * University of Florida ** gwerdos-at-gnv.ifas.ufl.edu * * Gainesville, FL 32611 ** * ********************************************************** ********************************************************** * Greg Erdos ** * * Director, ICBR EMCL ** Phone 904-392-1295 * * 218 Carr Hall ** FAX 904-392-8598 * * University of Florida ** gwerdos-at-gnv.ifas.ufl.edu * * Gainesville, FL 32611 ** * **********************************************************
} From: IN%"KARUZIS-at-wccf.mit.edu" "GLENN HOLM" } Subj: RE: U.S. supplier for vertical staining dishes? } } Return-path: {KARUZIS-at-wccf.mit.edu} } Received: from WCCF.MIT.EDU by gnv.ifas.ufl.edu (PMDF V4.2-11 #3240) id } {01HC7V1A7RVK8X8BBU-at-gnv.ifas.ufl.edu} ; Wed, 11 May 1994 15:38:11 EST } Received: from wccf.mit.edu by wccf.mit.edu (PMDF V4.2-14 #2603) id } {01HC7UYC8CK48WXF5V-at-wccf.mit.edu} ; Wed, 11 May 1994 15:37:26 EST } Date: Wed, 11 May 1994 15:37:26 -0500 (EST) } From: GLENN HOLM {KARUZIS-at-wccf.mit.edu} } Subject: RE: U.S. supplier for vertical staining dishes? } To: GWERDOS-at-gnv.ifas.ufl.edu } Message-id: {01HC7UYC8CK68WXF5V-at-wccf.mit.edu} } Organization: Mass. Inst. Tech. - Whitaker College } X-VMS-To: IN%"GWERDOS-at-gnv.ifas.ufl.edu" } MIME-version: 1.0 } Content-transfer-encoding: 7BIT } } greg- } } thanks for the reply. i looked at the Fisher staining dish, but what we } want is a bit larger and holds the slides vertically in a rack, not in } grooves in the side of the dish like a coplin jar. will keep hunting. } ------------------------------------------------------------------ } |Glenn Holm Internet:karuzis-at-wccf.mit.edu| } |M.I.T Dept. of Brain + Cog. Sci. This VAX doesn't do NeXTmail | } |Cambridge, MA 02139 "Real Neuroscientists don't do gels!" | } ------------------------------------------------------------------
############%%%%%%%%%%%%%%%%##############%%%%%%%%%%%%%%########## I just remembered where I saw it. Shandon & Lipshaw Catalogue. Purveyors of goods for pathology, cytology and mortuary. Call 800-547-7429 and they will send a catalogue that smells like a morgue. Check out page 184 for a rack that holds 38 slides vertically on end.
Or if you are close to a hospital pathology lab or autopsy lab they may have the catalogue.
A few years ago, I used a very nice silver enhancement kit from Janssen called IntenseII. I used it only for light microscopy, but I know that people have used it successfully for EM as well. It was simple and quite forgiving with regard to development time. I also believe the reaction can be slowed by diluting the reagents. I don't remember who the US distributor for Janssen is now, but for some reason I'm thinking it might be Vector.
Cheers
David Morilak Dept Pharmacology UT Health Science Center San Antonio morilak-at-uthscsa.edu
Dr. Avalos, I lost your E-mail address. Please send it. Thank you. My apologies to the others on the list. Dr. Stanley L. Flegler flegler-at-pilot.msu.edu
I can't afford a new scanner but the thought of upgrading my Hitachi S450 with the Semicaps imaging system has excited me. My questions are...does anyone have the system on an analog scope? How does it grab the image off the scope? Can I use it with my TEM also? Is it as good as the other systems out there?
John Grazul Rutgers University Electron Microscope Facility
I saw a request recently from Michael Winton concerning TEM sample preparation of silicon on sapphire, but I saw no responses posted to the listserver.
Did anyone out there have a good (or reasonably good!) answer for him?
Thanks!
David Henriks South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
Now I remember. The comercial daylight kits for silver enhancement require the addition of gum arabic in order to slow the reaction. I can't immediately recall the deatails but maybe it will ring a bell in a younger brain ********************************************************** * Greg Erdos ** * * Director, ICBR EMCL ** Phone 904-392-1295 * * 218 Carr Hall ** FAX 904-392-8598 * * University of Florida ** gwerdos-at-gnv.ifas.ufl.edu * * Gainesville, FL 32611 ** * **********************************************************
} Does anyone have proven sample preparation techniques for thin film } silicon on sapphire substrates?
Hi, Michael:
I tried to post this mail last weekend but the listserver was down. I hope this can be any help to you.
I made such a sample once. It came out beautifully. I assume that you are talking about cross-sectional TEM sample because it is easy to make plane-view sample. The recipe is as following:
1. Stack two pieces face to face together with a very thin layer of M-bond. Curing samples at 170 degree C for two hours. 2. Grind and polish one side. (with SiC paper first then finish with diamond paste). The thickness of the sample is 100 um now. 3. Dimple the other side to less than 25 um. 4. Ion mill the sample.
The experiences of grinding, polishing and using dimpler are necessary. Since sapphire is brittle and hard, you need patience. Good luck. -- Tan-Chen Lee Cornell University tanchen-at-msc.cornell.edu
Dear Clayton Smith: Thank you for your letter of May 4, 1994 and the catalog. Dr. Curzon will make a decision. My apologies to the others on the list. Sandy Burany
Subject: Time:1:49 PM OFFICE MEMO N2 from LN2 Date:5/12/94 In the recent discussions about using dry nitrogen to backfill vacuum systems, several of us have mentioned collecting the gas that boils off liquid nitrogen storage vessels and using it for this purpose. There is a very inexpensive way to do this that I devised some 20 years ago and used for many years with good success. What you do is to run a flexible, non-collapsible plastic tube (I prefer polyethylene tubing to Tygon, because Tygon seems to exude so much plasticizer - which might find its way into the vacuum system) from the liquid nitrogen container to the gas inlet of the vacuum system. At a convenient site, insert a tee joint into this tube. A large, flexible, inflatable, plastic container (a beach ball or childs toy animal) is attached to this tee joint by means of a length of soft, highly-flexible surgical rubber tubing. Before installing this tubing, however, take a sharp scalpel or razor blade and make a clean slit about 100 mm long in it. This slit will ordinarily close tightly enough so that the nitrogen from the storage vessel will flow into the plastic ball. When the ball becomes full, however, the slit will serve as a primitive pressure release valve by opening slightly to allow the gas to escape, thereby preventing the ball from rupturing. When the gas inlet is opened, the dry nitrogen in the ball will flow into the system under atmospheric pressure, and so there is no danger of over-pressurization. A small weight can be placed on the ball to sustain flow after the system reaches atmospheric pressure, if desired. A ball that is a half meter in diameter will store enough gas to fill most laboratory systems several times. This is a very inexpensive arrangement, but one that works quite well, and an inflated dinosauer that is hooked to your SEM will provide a topic of conversation for everyone who visits your lab. Again I mention that this technique, and many others, are described in my recent book on Vacuum Methods in Electron Microscopy, which can be ordered from Portland Press Ltd., c/o Ashgate Publishing Co, Old Post Road, Brookfield, VT 05036-9704, USA. (Fax: 802-276-3837); or Portland Press Ltd., Commerce Way, Colchester, CO2 8HP, UK (Fax: 0206-799331)
check Y_D Stierhof, et al J of electron microsc tech 17:336 for a nice comparison of different silver enhancement techniques
------- Forwarded Message Follows - - - - - - -
Now I remember. The comercial daylight kits for silver enhancement require the addition of gum arabic in order to slow the reaction. I can't immediately recall the deatails but maybe it will ring a bell in a younger brain ********************************************************** * Greg Erdos ** * * Director, ICBR EMCL ** Phone 904-392-1295 * * 218 Carr Hall ** FAX 904-392-8598 * * University of Florida ** gwerdos-at-gnv.ifas.ufl.edu * * Gainesville, FL 32611 ** * **********************************************************
Hello We're looking at living cultured cells in a perfusion bath maintained at 37 deg C. To minimise heat loss through the objective lens, we heat the lens during the experiments. I'm looking for a source of an appropriate immersion oil that does not change refractive index over the range 20-40 deg C to reduce any abberations in images of the cells. Any suggestions would be appreciated. Thanks Malea
************************************************************************ Malea Kneen Department of Physiology The University of Melbourne Grattan Street, Parkville, 3052 Victoria, AUSTRALIA
Re: Donald Grimes request for microscopy software details:
There are four PC packages published by the UK Institute of Materials,
titled
Scanning Electron Microscopy by John Humphreys Transmission Electron Microscopy by Peter Goodhew Electron Diffraction by Peter Goodhew Analysis in the EM by Peter Goodhew, John Humphreys and Graham Cliff
There is also a nice package on Stereographic Projection by John Humphreys, which is useful for those working with crystals.
They are all intended for introductory use (for people new to EM) and employ quite a lot of graphics. They are NOT research tools for competent microscopists. We use them with 2nd and 3rd year undergrads and people attending short courses on EM.
Each package costs about US$145. They are distributed by
Ashgate Publishing Co Old Post Road Brookfield VT 05036 USA Tel 802 276 3162 Fax 802 276 3837 (for the world except Europe)
and
Institute of Materials 1 Carlton House Terrace London SW1Y 5DB Tel 071 976 1338 Fax 071 839 2078 (for UK and Europe)
Work is in progress to upgrade these programs from DOS to Windows versions but there are no immediate plans for Mac versions (unless someone would like to volunteer to translate/re-emgineer them).
Peter Goodhew (series editor) University of Liverpool goodhew-at-liv.ac.uk (44) 51 794 4665 Fax (44) 51 794 4675
How are publication quality trace analysis results generated? I can get a stereogram with all the poles etc plotted on a computer, but how do you go about drawing the great circles (which are best done using a Wulff net} ? Some Bezier curves can be drawn to get the right curvature etc. but this would be pretty inaccurate, generally. Drawing by hand, unless done by a professional is not publication quality. Any suggestions? Thanks, Sharath dakshs-at-rpi.edu
Message-ID: {MAILQUEUE-101.940513102236.352-at-bmg.bhs.uab.edu} To: microscopy-at-anlemc.msd.anl.gov
Don, Signal Analytics Corporation makes several exellent digital imaging packages sold under the name IP Lab Spectrum for the Macintosh. Their forte is that the software is easy to install, configure, and use and the price is relatively (?) inexpensive, their base package starting around $1500. We have such a system up and running in our digital imaging facility at the University of Alabama at Birmingham and have had great success with it. Their address is:
Signal Analytics Corporation 440 Maple Avenue East Suite 201 Vienna, Virginia 22180
Phone 703-281-3277 Fax 703-281-2509 They are not on Internet yet but anticipate entering the network in the near future. I am currently working with the company to set up a user's group archive that will be accessable via Internet using Gopher. We hope to have this up an working in the next month or two. Any input from any interested parties would be greatly appreciated
Kevin McCarthy Kevin McCarthy Assistant Professor Department of Cell Biology University of Alabama at Birmingham Birmingham, Alabama 35294 Phone 205-934-9923/9924 Fax 205-934-7029
Message-Id: {MAILQUEUE-101.940513084352.998-at-prl.pulmonary.ubc.ca} To: microscopy-at-anlemc.msd.anl.gov
To Malea Kneen could you please repeat the last part of your message about an appropriate immersion oil. For some reason I did not recieve the complete message so am not sure what type of oil you are looking for. Thanks Mark Elliott, UBC-Pulmonary Research Lab Vancouver, Canada
As many of the readers of this list have funding grants from DoE(Energy & Water Budget), NSF, and NASA (VA-HUD-IA Budget) I thought that you might want to know about this gem which is going through the US Congress.. It's not microscopy, however, it will likely impact our research funding. Apologizes in advance if you've already seen this information, or if you are one of our International Subscribers who isn't interested in the US research budget.
Nestor J. Zaluzec ANL EMCenter
---------- WHAT'S NEW by Robert L. Park Friday, 13 May 94 Washington, DC
1. BUDGET: CONGRESS TURNS OFF THE LIGHT AT THE END OF THE TUNNEL. Yesterday, the House Appropriations Committee divvied up the FY 95 budget among the 13 Subcommittees. Only Military Construction was up from last year. Energy and Water came out $81M below the President's request. VA-HUD-IA, which includes both NASA and NSF, came out $361M below the President's request; Subcommittee chair Louis Stokes (D-OH) said he had "grave concerns about whether this allocation can fund the space station." The specter of the SSC hangs over the space station debate; supporters of the space station like to point out that the money saved from the SSC last year did not go to science. Actually, no one in their right mind expects savings from the space station to go to science either-- the important thing is stop the money from going the other way.
Alexey Sidorenko phone: (7-095) 932-88-61 Moscow State University E-mail: avs-at-srdlan.npi.msu.su Institute of Nuclear Physics http://www.npi.msu.su/people/avs/avs.html
I just got my hands on the softcover book "The internet Companion-A beginner's guide to global Networking", and recommend it highly to anyone who is not a computer expert. Even though, I used the internet now for over two years, not until browsing through the text, did I finally understand some of the things I was doing without knowing why. If everyone in the userlist were to read the book, we would probably not get certain message-types that are explicitly discouraged in chapter three of the book.
***** ************ ************** ************ *Cesar D. Fermin, Ph.D |Fax (504) 587-7389 *Tulane Medical School |Answ. Mach.(504) 584-2618 *Pathology/SL79 |Secretary (504) 584-2436 *New Orleans, La 70112 | Lab (504) 584 2521 ***** ***************** ***********************
Message-Id: {MAILQUEUE-101.940516144653.198-at-prl.pulmonary.ubc.ca} To: microscopy-at-anlemc.msd.anl.gov
Does anyone know of a similar network to the microscopy one that deals with tissue culture problems???? We are having problems with guinea pig lymphocyte stimulation with phytohemaglutinin and need advice. Any help would be appreciated. Thanks Mark Elliott Pulmonary Research LAb Vancouver Canada
Have you checked out the "Big Dummies Guide to the Internet"? It is a Hypercard stack which covers most aspects of using the Internet. It is available via ftp. I think I got my copy from sumex-aim.stanford.edu.
Glen MacDonald Hearing Development Laboratories RL-30 University of Washington Seattle, WA 98195 (206)543-8360 glenmac-at-u.washington.edu
On 16 May 1994, Fermin, Cesar wrote:
} } Hello: } } I just got my hands on the softcover book "The internet Companion-A beginner's } guide to global Networking", and recommend it highly to anyone who is not a } computer expert. Even though, I used the internet now for over two years, not } until browsing through the text, did I finally understand some of the things I } was doing without knowing why. If everyone in the userlist were to read the } book, we would probably not get certain message-types that are explicitly } discouraged in chapter three of the book. } } ***** ************ ************** ************ } *Cesar D. Fermin, Ph.D |Fax (504) 587-7389 } *Tulane Medical School |Answ. Mach.(504) 584-2618 } *Pathology/SL79 |Secretary (504) 584-2436 } *New Orleans, La 70112 | Lab (504) 584 2521 } ***** ***************** *********************** }
Mark Elliott queries about a similiar discussion group about tissue culture. In looking through a list of numerous listservers/discussion groups out their I can up with this: MEDCC-L-at-UAFSYSB Issues related to the study of media, culture, etc. Hope this helps.
I'm sorry for my annoying message to this mailing list. I wanted to subscribe and expected to get an automatic response with information.
Thanks to everyone who helped me.
Alexey
Alexey Sidorenko phone: (7-095) 932-88-61 Moscow State University E-mail: avs-at-srdlan.npi.msu.su Institute of Nuclear Physics http://www.npi.msu.su/people/avs/avs.html
you might try any of the bio. or sci. news groups. Reading "news" can be a real chore, and you have to wade through a lot of noise, but you do find the occasional gem. There are a handful of bionet news groups that function more or less like our mail lists, ie posting of questions and answers that all interested parties get to "eavesdrop" in on. I believe you can ftp a utility called TheNews from sumex-aim which holds your hand as you custom,ize your newsgroup subscriptions and set up your sessions. It also gives a listing of the zillion or so groups that are available. Good luck.
David Morilak UT Health Science Center Dept Pharmacology San Antonio morilak-at-uthscsa.edu
I have just tried to access sumex-aim.stanford.edu to obtain a copy of "Big Dummies Guide to the Internet", but the server is not available. Does anyone know of another ftp site?
_____________________________________________________________________ | James V. Jester | Dept Ophthalmology | | jester-at-crnjjsgi.swmed.utexas.edu | UT Southwestern Medical Center | | (214)648-7215 | 5323 Harry Hines Blvd | | | Dallas, TX 75235-9057 | |__________________________________|__________________________________|
Hi there, I am sorry about not sending the complete reference for the book. I hope that the following is of help.
RE:Internet Companion-Laquey
Source for Internet comp-Laquey The publisher is Addison-Wesley Publishing Co. ISBN 0-201-62224-6 Fourth Printing March 1993 ($11.00)
However, the text can be downloaded by anonymous FTP from FTP.STD.COM. If you have a Word Wide Web (WWW) browser like Mosaic you can get it from many sources, including our here at Tulane, by looking in the category internet information and is free, I like the book because is a nice ready reference. Good luck.
P.D. Other commonly recommended texts are also available from the same source.
Just received a demo disk for the Electron Flight Simulator program from Small World. It looks interesting. Does anyone have any experience with, or comments about this product? They're asking $449. Is it worth it?
.............................................................................. John Hudak hudakjm-at-mcmaster.ca I.M.R. - Electron Optics (905) 525-9140 Ext.24890 ABB Rm. 331 FAX: (905) 521-2773 McMaster University 1280 Main St. West Hamilton, Ont. L8S 4M1 Canada
Does anyone know of a commercial source for various thin ceramic (150 - 300 angstrom thick) films on salt flats (1" x 1" minimum area)? We need these for some "non-microscopy" experiments we are considering. The compositions required are SiO, SiO2, and/or SiN. Non-stoichiometric films are OK, we only need the average molecular weight to be correct.
Of course, we need these yesterday! Thanks to all in advance.
Regards, Bill Lamberti.
William A. Lamberti Office LA-196 Exxon Research and Engineering Company Route 22 East Annandale, NJ 08801 (908)730-2144 office (908)730-2262/2104 labs (908)730-3042/3051 fax Email: walambe-at-erenj.com
John I recieved the same package yesterday. The program does indeed look useful, but I have a serious problem with the price. $500 seems like a lot to pay for a nice interface, especially when non-Window versions are available free on the EMMPDL. Just my two cents. ********************************************************************** * Glenn Poirier glenn_p-at-geosci.lan.mcgill.ca * * Electron Microprobe Lab Phone: (514) 398 6774 * * Earth and Planetary Sciences Fax: (514) 398 4680 * * McGill University THERE ARE THREE SIDES TO EVERY STORY; * * Montreal, Quebec YOUR SIDE MY SIDE AND THE TRUTH * **********************************************************************
I am shopping for a new SEM and wanted to know what active users would recommend or not recommend. Ideally, I need an actual resolution of 75 A at 25K magnification. Our expected budget will be $200,000 and we would also like a microprobe. This microscope will be multi-departmental (biological sciences and chemistry). Thank you in advance, Page Owen
Page Owen Dept. of Botany Connecticut College 270 Mohegan Ave., Box 5555 New London, CT 06320 (203)439-2147
SPECIAL ISSUE - ELECTRON SPECTROSCOPIC IMAGING AND ANALYSIS TECHNIQUES. A SELECTION OF PAPERS FROM THE 4TH EUROPEAN WORKSHOP HELD AT THE INSTITUTE OF PHYSICS, UNIVERSITY OF MUNSTER, GERMANY
Journal of Microscopy, Vol. 174, Pt. 3, June 1994, pp. 133 - 142.
KOHLER ILLUMINATION IN THE TEM: FUNDAMENTALS AND ADVANTAGES
G. BENNER & W. PROBST Electron Optics Division, Carl Zeiss, 73446 Oberkochen, Germany
Summary
Kohler illumination is the most favourable design for the illumination path of an electron microscope with a condenser objective lens. The new illumination system of the EM 910 and EM 912 OMEGA allows both wide area (Kohler) illumination for TEM operation and spot illumination for analytical investigations. Compared to conventional systems and objective lenses with a condenser mini lens, this system offers many advantages. In addition to the homogeneous, highly coherent and parallel illumination of every point in the specimen, it offers advantages for selected area diffraction and spot scan mode. Combined with the electron optical selection of a condenser aperture, this illumination system provides the flexibility necessary to achieve optimum illumination for the specimen.
Journal of Microscopy, Vol. 174, Pt. 3, June 1994, pp. 143 - 148.
INFLUENCE OF ZERO-LOSS FILTERING ON ELECTRON OPTICAL PHASE CONTRAST
P. HIRSCH & L. REIMER Physikalisches Institut, Universitat Munster, Wilhelm-Klemm- Strasse 10, 48149 Munster, Germany
Summary
Complex scattering amplitudes are used to calculate the phase contrast of colloidal gold particles.Comparison of measurements of the phase contrast intensity at the centre of the gold particle as a function of defocus for unfiltered and zero-loss filtered images demonstrates the increase in phase contrast achieved by zero-loss filtering even for a thick carbon substrate film. The granulation of amorphous germanium films is measured by the spatial root mean square values of image intensity in a defocus series.
Journal of Microscopy, Vol. 174, Pt. 3, June 1994, pp. 149 - 159.
IMAGE-EELS: SIMULTANEOUS RECORDING OF MULTIPLE ELECTRON ENERGY- LOSS SPECTRA FROM SERIES OF ELECTRON SPECTROSCOPIC IMAGES
K.-H. KORTJE Institute of Zoology, University Hohenheim, Garbenstrasse 30, D- 70593 Stuttgart, Germany
Summary
A new approach for element microanalysis with energy-filtering transmission electron microscopy (EFTEM) is presented which was accomplished with the CEM 902 electron microscope (Zeiss, Germany). This method is called Image-EELS, because it is a synthesis of electron energy-loss spectroscopy (EELS) and electron spectroscopic imaging (ESI). Series of energy-filtered images at increasing energy losses are recorded from one area with a TV camera. In a second step the intensity of selected regions in the image stack is measured with an image analysis system and plotted as a function of the energy loss. Thus many spectra from different objects can be calculated from one image series and compared with each other. The spatial resolution of EELS is considerably enhanced, the noise is decreased because many pixels from irregular objects are integrated, and the information from ESI can be analysed as a function of the energy loss.
Journal of Microscopy, Vol. 174, Pt. 3, June 1994, pp. 161 - 169.
OPTIMAL CONDITIONS FOR THE USE OF CORRESPONDENCE ANALYSIS FOR ELEMENT DETERMINATION IN EELS IMAGES
E. S. GELSEMA, * A. L. D. BECKERS* & W. C. DE BRUIJN + * Department of Medical Informatics, and + AEM-unit Pathological Institute, Erasmus University, PO Box 1738, 3000 DR Rotterdam, The Netherlands
Summary
Correspondence analysis is used for the determination of element localization and element concentration in electron energy-loss imaging. The introduction of a new method for the quantification of element concentration embedded in the protocols of correspondence analysis may be regarded as a useful replacement of the power-law estimation technique, especially in regions of the spectrum where the latter method cannot be used. Conditions for the optimum use of the method are established.
Journal of Microscopy, Vol. 174, Pt. 3, June 1994, pp. 171 - 182.
QUANTITATIVE ELECTRON SPECTROSCOPIC IMAGING IN BIO-MEDICINE: METHODS FOR IMAGE ACQUISITION, CORRECTION AND ANALYSIS.
A. L. D. BECKERS,* W. C. DE BRUIJN,+ E. S. GELSEMA,* M. I. CLETON-SOETEMAN# & H. G. VAN EIJK# *Department of Medical Informatics, +AEM-unit Pathological Institute, #Department of Chemical Pathology, Erasmus University Rotterdam, P.O. Box 1738, 3000 DR Rotterdam, The Netherlands.
Summary
Many questions about the metabolism of specific elements in the human body might be answered if elemental concentrations could be measured in situ in cells. With electron energy-loss spectroscopic imaging (ESI), concentrations can potentially be determined with high spatial resolution. The theory of the quantification procedure has already been derived. Many practical instrument-related problems, however, have to be solved. In the current research an energy-filtering TEM is used and the image-acquisition chain is examined in detail. Quantification requires images to be recorded over a large dynamic range. To solve this problem, the use of optical attenuation filters has been introduced. The use of the combination of a scintillator screen and a TV-camera as a detection system has consequences for the processing of the data. Corrections for the camera photometric sensitivity and , to some extent, for shading are necessary. Further consequences of such a detection system for the correction of the element-aspecific spectral background and element detection are discussed. The derived methodology is tested in several ways and finally applied for the quantitative analysis of iron in liver parenchymal cells of a porphyria cutanea tarda patient.
Journal of Microscopy, Vol. 174, Pt. 3, June 1994, pp. 183 - 193
EFFECT OF SPECIMEN THICKNESS AND CARBON CONTENT ON THE CARBON PEAK REGION IN EEL SPECTRA
R. DOOR, K. D. HABERLE & R. MARTIN Sektion Elektonenmikroskopie, Universitat Ulm, D-89069 Ulm, Germany
Summary
Amorphous carbon films of different thicknesses and combinations of carbon films with CaF2 films of different thicknesses were used to create a standard reference library of EEL spectra. The relationship between the size and shape of the carbon peak region and the low-loss part of the spectrum was determined. The steepness of the background of the C-K edge, the size of the carbon peak and the plasmon shoulder on top of the carbon peak were investigated as a function of the thickness of the films and of the relative amount of carbon after normalization of the zero- loss peak. The minimum at the C-K edge and the maximum of the carbon peak were considered as examples for a modelling of the carbon peak region in different specimens. Methodological aspects which affect the accuracy of the measurement (characteristic lines of the detector, calculation of specimen thickness, method of t/lamda measurement, mass loss, energy resolution) were examined. The results indicate that modelling of the whole carbon peak region, considering its dependence on mass thickness and carbon concentration, should be possible from our standard reference spectra without use of the low-loss region. The use of modelled standard reference spectra for background extrapolation in biological specimens with unknown calcium content is proposed.
Journal of Microscopy, Vol. 174, Pt. 3, June 1994, pp. 195 - 206.
APPLICATION OF RECORDING AND PROCESSING OF ENERGY-FILTERED IMAGE SEQUENCES FOR THE ELEMENTAL MAPPING OF BIOLOGICAL SPECIMENS: IMAGING-SPECTRUM
JEAN-LOUIS LAVERGNE, * + COLETTE FOA, # PIERRE BONGRAND, # DOMINIQUE SEUX ++ & JEAN-MICHEL MARTIN* * Ecole Centrale de Lyon, Laboratoire de Tribologie et Dynamique des Systemes, URA CNRS 855, Departement de Technologie des Surfaces, F-69131 Ecully Cedex, France + KODAK-PATHE, Laboratoire d'Analyses, Z.I. nord, F-71102 Chalon sur Saone, France # INSERM U 387, Laboratoire d'Immunologie, Hopital St Marguerite, F-13277 Marseille Cedex 9, France ++ Laboratoire de Developpement et de Pathologie des Tissus Dentaires, UPR CNRS 412, Faculte d'Odontologie, F-69372 Lyon Cedex 8, France
Summary
Computerized energy-filtered transmission electron microscope (EFTEM) permits the recording and the processing of energy- filtered images, allowing a part of an electron energy-loss spectrum for each picture element to be obtained. This method, called 'Imaging-Spectrum', uses a Zeiss CEM902 coupled to several image analysis systems. The actual configuration records sequences of 48 images, 256 x 256 pixels, in steps of the energy loss, žE . Processing these sequences results in part of a core- loss EELS-spectrum for each pixel. This approach produces elemental maps with a short processing time. We have implements three kinds of background calculation for the image subtraction. The influence of the irradiation dose and of the energy selecting slit width on the quality of the spectra is investigated. The method is applied to the analysis of some biological specimens (pericellular coat behaviour during adhesion between macrophages and red blood cells and location of calcite microcrystals in dental pulp cells). The Imaging-Spectrum method appears to be suitable for the analysis of large areas.
Journal of Microscopy, Vol. 174, Pt. 3, June 1994, pp. 207 - 223.
EVALUATION OF LANTHANIDE TRACER METHODS IN THE STUDY OF MAMMALIAN PULMONARY PARENCHYMA AND CARDIAC MUSCLE BY ELECTRON ENERGY-LOSS SPECTROSCOPY
H. FEHRENBACH, A. SCHMIEDL, F. BRASCH & J. RICHTER Abt. Elektronenmikroskopie, Zentrum Anatomie, Kreuzbergring 36, D-37075 Gottingen, Germany.
Summary
Lanthanum (La) has widely been used as a tracer to study the integrity of plasma membranes. With conventional transmission electron microscopy (cTEM), the absence of electron scattering deposits from the cytoplasm has generally been assumed to reflect an intact cell membrane. However, the application of electron spectroscopic imaging (ESI) and electron energy-loss spectroscopy (EELS) reveals that electron scattering deposits may be present which do not contain La. However, La could be detected in regions of pulmonary parenchyma and cardiac muscle that were devoid of electron scattering deposits. Therefore, to exclude misinterpretations based on CTEM the application of microanalytical techniques is strongly recommended for the study of the integrity of plasma membranes by means of La tracers. In addition, ESI and EELS are shown to distinguish between different tracers in simultaneous applications of La and terbium (Tb) which were used at the different faces of the pulmonary air-blood barrier. The analysis of the distribution of both tracers which form electron scattering deposits, indistinguishable by cTEM, may help us to understand the different functional significance of cellular alterations of both cellular borders of the barrier. As was shown for La, however, strictly controlled conditions are mandatory during the fixation procedure because an increase in the incubation time to more than 1 h in samples of pulmonary parenchyma may result in the occurrence of La deposits within the cytoplasm. In the absence of electron scattering deposits, the presence of La in glycogen granules and ribosome-containing areas of various types of alveolar septal cells even after 15 min incubation indicates that the absence of deposits does not necessarily correspond to the absence of the tracer.
Journal of Microscopy, Vol. 174, Pt. 3, June 1994, pp. 225 - 232.
DEMONSTRATION OF ALUMINIUM IN POLYPHOSPHATE OF LACCARIA AMETHYSTEA (BOLT. EX HOOKER) MURR. BY MEANS OF ELECTRON ENERGY- LOSS SPECTROSCOPY
I. KOTTKE * & F. MARTIN + * Universitat Tubingen, Botanishches Institut, Spezielle Botanik und Mykologie, Auf der Morgenstelle 1, D-72076 Tubingen, Germany + INRA, Laboratoire de Microbiologie Forestiere, Centre de Recherches Forestieres, 54280 Champenoux, France
Summary
Toxic aluminium species in acidified soil damage the rootlets of trees. The symbiotic association with ectomycorrhizal fungi may, however, protect the rootlets by sequestration of aluminium in polyphosphates. Electron energy-loss spectroscopy was used to identify the relative amount of aluminium in the polyphosphate of the mycelium of the ectomycorrhizal fungus Laccaria amethystea. A three-window method at the L-2,3 edge yielded results which were in agreement with the spectra obtained at the aluminium K edge. The aluminium net distribution images were recorded at 85 eV, the backgrounds at 70 eV and 60 eV, setting the energy selecting slit to 5 eV and the beam current to 15 microA. The method can be used to study the sequestration of aluminium in mycelia of ectomycorrhizal fungi under different stress situations.
Journal of Microscopy, Vol. 174, Pt. 3, June 1994, pp. 233 - 238.
ESI IN SITU ANALYSES OF EXTRACHROMOSOMAL RNP GRANULES
S. ABOLHASSANI-DADRAS, * G. H. VAZQUEZ-NIN, *# O. M. ECHEVERRIA, *# V. BOUTINARD ROUELLE-ROSSIER* & S. FAKAN* * Centre of Electron Microscopy, University of Lausanne, 27 Bugnon, CH-1005 Lausanne, Switzerland # Laboratory of Electron Microscopy, Department of Biology, Faculty of Sciences, National Autonomous University of Mexico, Mexico City, D.F. 04510, Mexico
Summary
Observation of unstained ultrathin sections of salivary gland cells of Chironomus thummi and C. tentans, by means of electron spectroscopic imaging (ESI), has revealed that phosphorus is distributed in two types of granular structures in the nucleoplasm of these cells. In addition to a specific type of premessenger ribonucleoprotein (RNP) particle, known as the Balbiani ring (Br) granule, ESI revealed a new type of phosphorus-rich small granular component. Examination of unstained sections by energy-filtering transmission electron microscopy (EFTEM) offers the opportunity of obtaining the signal from the specimen alone, thus avoiding the possible contributions of heavy metals present in any staining product.
I need a reviewer for a short manuscript on the use of electron microscopy in the examination of minerals and soils. If any subscribers are qualified and willing to help in this matter please contact me directly.
*************************************** David Garrett "DGARRETT-at-GAB.UNT.EDU" University of North Texas Dept. Biological Sciences (817)565-3964 Fax (817)565-4136 ***************************************
A recent posting listed the FTP site at STD.COM for obtaining the Internet Companion. It would be a great help if the posting individual could provide pointers to the directory where it is listed. The search engines always seem to be tied up when I try to log on, so this would be a help. Thanks.
Does anybody have any information regarding the thickness and roughness of nitrocellulose in amyl acetate (0.1%)? The procedure we used was to take 2 drops of the solution in a pasteur pipet and cast onto a very clean glass coverslip. This technique is used for a motility assay of actin filaments on myosin absorbed on the nitrocellulose layer. Any references or protocols would also be helpful.
Margaret (Peggy) Bisher
NEC Research Institute, Inc. 4 Independence Way Princeton, NJ 08540
Subject: Time:3:32 PM OFFICE MEMO LM Date:5/19/94 I am doing basic morphological studies on rat kidneys. I am looking at 1 micron epon sections that have been processed for standard electron microscopy. I have a few questions on what I am looking at, and I am looking for someone that I can talk to that might be able to answer some simple questions such as what kind of artifacts will you get when you perfuse, or is a perfused kidney even preferred over a biopsy, how much variation is there from rat to rat, etc., etc.. If anyone has experience working with rat kidney, I would really appreciate it if you could contact me. Thank-you, Jeanne
There is a new Macintosh software for electron diffraction and crystallographic calculations developed by Drs. J. M. Zuo and H.R. Zhu. Preliminary report can be found in last year MSA proceedings. This program plots crystal structure, stereoprojection electron diffraction pattern (SAD and CBED) and backscattered kikuchi pattern with a simulated microscope control panel. Intensity of kinematic approximation is used. The program also has calculator for complex numbers and vectors of real and reciprocal space, and functions for calculating electron and x-ray structure factors and Bragg angles etc. For more information contact
We have an Akashi/ISI EM-002A TEM (1988), one of only eight or nine made. I am keen to get in touch with any other owners of these instruments with regards to sorting out technical problems and obtaining spare parts. This would include owners of the EM-002B, which is similar in many ways. Yours faithfully,
Richard Easingwood EM Unit,School of Medicine University of Otago Dunedin New Zealand
Richard Easingwood Department of Anatomy and Structural Biology, P.O. Box 913 University of Otago, Dunedin, New Zealand. Fax:64-3-479 7254 Telephone:64-3-479 7301
I recently downloaded the Internet Companion successfully, as follows:
site: ftp.std.com dir: \ftp\OBS\The.Internet.Companion files: internet.companion (349 910 bytes) (Text version) internet.companion.Z (zipped version, Unix) COPYRIGHT Order.form Tracy.gif (I assume a .gif of the author, Tracey Laquey)
Note: if you are downloading to a DOS machine, you will have to save the files to a DOS-acceptable filename..(eg internet.txt) The ftp-site filename of {internet.companion} is OK in Unix, but not in DOS. [The same applies if you ever download uuencoded or zipped files. If the filename within the uuencoded file is more than 8 characters or contains unnacceptable characters, the file may not be uudecoded on a DOS machine. It is then necessary to change the filename within the compressed file (using any text editor) before uudecoding etc.]
Hello Ca imagers! We have been trying to image Ca transients in some cells (neutrophils and some mouse macrophages) following cross-linking of surface receptors but we have been running into some difficulties. Our hardware configuration is as following: We are using an intensified Hamamatsu CCD camera mounted on the side port of a Zeiss Axiovert. We are using the following filters: excitation 340nm 10nm FWHM or 350nm 10nm FWHM dichroic 375nm emission 405nm 45nm FWHM 490nm 45nm FWHM
The objective is a Zeiss 40X UltraFluar. We also use OD filters in the excitation path to minimize bleaching of the probe (usually 25 or 50% transmission). The dynamic range of the measurement, in terms of the ratio between the Ca-free and Ca-bound ratios, is only ~10, which may be insufficient to give good accuracy at Ca concentrations away from the Kd of the probe! This problem is compunded by the 8-bit accuracy of the imaging system (Image 1/FL, Universal Imaging Corp. using a Matrox video board). Does anybody have comments? Since most of the papers report values of [Ca]i and not the raw ratio values, I cannot judge whether we are doing something wrong! Is this a common observation? Comparable measurements with a spectrofluorometer on cells in suspension give us a dynamic range (as defined above) of ~40. Is anybody willing to share her/his experience? Any comments/suggestions will be greatly appreciated.
Thank you in advance!
************************************ *Massimo Sassaroli * *Dept. of Physiology and Biophysics* *Mount Sinai School of Medicine * *New York, NY 10029-6574 * *Tel: 212-241-9512 * *sassaroli-at-msvax.mssm.edu * ************************************
P.S. : Is there any other interest group dealing specifically with Ca measurement techniques? Thanks again
Hello, Last week the results of a poll on how people record their images was posted. After briefly scanning the replies, I deleted the file. Now I find myself wishing I hadn't. Could someone email the file to me? Is there an archival site where I could retrieve it from (seems to me I saw this listed here before). The reason I would like to recover this file is that we are contemplating interfacing a PC with our Hitachi S-4000 FESEM. Anyone who has any suggestions, please reply.
___________________________________________________________________ Randy Nessler rnessler-at-uiowa.edu The University of Iowa Central Electron Microscopy Research Facility
We have an Akashi/ISI EM-002A TEM (1988), one of only eight or nine made. I am keen to get in touch with any other owners of these instruments with regards to sorting out technical problems and obtaining spare parts. This would include owners of the EM-002B, which is similar in many ways. Yours faithfully,
Richard Easingwood EM Unit,School of Medicine University of Otago Dunedin New Zealand
Richard Easingwood Department of Anatomy and Structural Biology, P.O. Box 913 University of Otago, Dunedin, New Zealand. Fax:64-3-479 7254 Telephone:64-3-479 7301
We have an Akashi/ISI EM-002A TEM (1988), one of only eight or nine made. I am keen to get in touch with any other owners of these instruments with regards to sorting out technical problems and obtaining spare parts. This would include owners of the EM-002B, which is similar in many ways. Yours faithfully,
Richard Easingwood EM Unit,School of Medicine University of Otago Dunedin New Zealand
Richard Easingwood Department of Anatomy and Structural Biology, P.O. Box 913 University of Otago, Dunedin, New Zealand. Fax:64-3-479 7254 Telephone:64-3-479 7301
Message-Id: {1994May23.141113.859926163-at-ms.sjdccd.cc.ca.us} To: microscopy-at-anlemc.msd.anl.gov (microscopy list)
I am trying to subscribe to the NIH-Image Listserver and was told that the address was listserv-at-soils.umn.edu and the message should be subscribe NIH-IMAGE {your name}
However when I did that I got a return receipt from listproc-at-soils.umn.edu that indicated that there was no NIH-IMAGE to subscribe to. Does anyone know the correct address for subscription. Thanks Judy Murphy San Joaquin Delta College Dept. of Microscopy Stockton, CA 209/474-5284 murphy-at-ms.sjdccd.cc.ca.us
The message, "No matches to nameserver query," is generated whenever the ph nameserver fails to locate either a ph alias or name field that matches the supplied name. The usual causes are typographical errors or the use of nicknames. Recommended action is to use the ph program to determine the correct ph alias for the individuals addressed. If ph is not available, try sending to the most explicit form of the name, e.g., if mike-fox fails, try michael-j-fox.
--------Unsent Message below:
Received: from anlemc.msd.anl.gov by nexus.uiowa.edu (1.38.193.4/931201) on Mon, 23 May 1994 23:41:13 -0500 id AA04787 with SMTP
Message-ID: {MAILQUEUE-101.940513102236.352-at-bmg.bhs.uab.edu} To: microscopy-at-anlemc.msd.anl.gov
Don, Signal Analytics Corporation makes several exellent digital imaging packages sold under the name IP Lab Spectrum for the Macintosh. Their forte is that the software is easy to install, configure, and use and the price is relatively (?) inexpensive, their base package starting around $1500. We have such a system up and running in our digital imaging facility at the University of Alabama at Birmingham and have had great success with it. Their address is:
Signal Analytics Corporation 440 Maple Avenue East Suite 201 Vienna, Virginia 22180
Phone 703-281-3277 Fax 703-281-2509 They are not on Internet yet but anticipate entering the network in the near future. I am currently working with the company to set up a user's group archive that will be accessable via Internet using Gopher. We hope to have this up an working in the next month or two. Any input from any interested parties would be greatly appreciated
Kevin McCarthy Kevin McCarthy Assistant Professor Department of Cell Biology University of Alabama at Birmingham Birmingham, Alabama 35294 Phone 205-934-9923/9924 Fax 205-934-7029
= = POSITION AVAILABLE = = Lab Manager for the Electron Microscopy Core Laboratory of the =Interdisciplinary Center for Biotechnology Research at the University =of Florida. = =DUTIES: Preparation and examination of biological specimens for = transmission and scanning electron microscopy in a facility that = serves the entire university community on a fee for service = basis. Collaborative research possible as part of the program. = Incumbent is expected to deal directly with the clients on = experimental design, execution of the project and preparation of = the data for publication, in consultation with the Scientific = Director of the laboratory. General lab management. = =QUALIFICATIONS: = experience in most aspects of biological electron microscopy. = Good communication skills, both oral and written. Ability to = deal on a one to one basis with investigators from a wide variety = of disciplines and backgrounds. = =STARTING DATE: Immediately = =APPLICATION DEADLINE: Open until position is filled. = =SALARY:$22,633 - 26,500. = =Inquiries by E-mail or full resumes to the address below = =********************************************************** =* Dr. Greg Erdos ** * =* Director, ICBR EMCL ** Phone 904-392-1295 * =* 218 Carr Hall ** FAX 904-392-8598 * =* University of Florida ** gwerdos-at-gnv.ifas.ufl.edu * =* Gainesville, FL 32611 ** * =**********************************************************
To: MICROSCO:CENTRAL cc: DRStadden:R_D:Armstrong Subj: AO Spencer Stereoscope ------------------------------------------------------------------ I am interested in knowing if anyone has used a repair service for AO Spencer stereomicroscopes. We have a vintage 1959 model with a detatched prism, and find it nearly impossible to repair ourselves, using makeshift alignment procedures. There must be a better way. If you know of a good repair service, or a method for doing it that we may not have thought of, or have gone through a similar experience, I'd love to hear from you. If you've seen this message before, I apologize, but I got several undeliverable mail notices when I sent it out about a month ago and received no replies. Please reply to: DRStadde+aCENTRAL%ARMSTRONG-at-MCIMAIL.COM or phone: (717)-396-5109 THANKS!
Hi I received the J. of Microscopy, June issue summary, but I have no clue on how it was encoded so that I may be able to decode it and read it!! My mail is received by a VAX but it is automatically rerouted to a SGI workstation and, finally, to a Mac running Eudora v. 1.4. The header of the message does not include the encoding method. Thank you for your help!
Dear Nestor, After posting to sci.microscopy about the poll, I realized that it was mailed to me personally by the person who conducted the poll. I guess the reason that I received the results was that I had participated in the poll. As far as the below message, it is all I recieved. Was a file to be included? I have the address of another contributor to the poll, so I will see if they kept a copy of the file. ___________________________________________________________________ Randy Nessler rnessler-at-uiowa.edu The University of Iowa Central Electron Microscopy Research Facility
On Mon, 23 May 1994 MICROARCHIVE-at-anlemc.msd.anl.gov wrote:
} Randy } } I'm not sure if I sent you what you wanted, but from your description } it was the only thing that looked right in the archives.... } } Nestor Zaluzec } ANL EMCentee } r
I was told that there is a new Polaroid product called 'Polapan 400' which is a medium-speed (ISO 400), coaterless, black-and-white professional film. The Polaroid company claims that this film is a new choice for T52 and T53 applications.
Does anyone have used this new film before? Can I get some comments from you? I use a SEM and take a lot of T53 films on rocks and minerals. Thanks.
Long Liang Electron Microprobe and SEM Lab ARCO Exploration and Production Technology 2300 West Plano Parkway Plano, TX 75075 E-mail : LLIANG-at-is.Arco.COM
The polaroid polapan 400 film works fine as a replacement for type 52, we use it with the same settings as the type 52. I don't know how they compare as far as cost, we were given 8-10 sheets to demo. I'm sure they will provide you a sample if you ask.
Does anyone have any experience with or sources for the electropolishing of indium alloys, specifically InCd or InPb? I need to prepare TEM specimens of these materials, and keep all the steps taken during preparation below room temperature. Any suggestions will be greatly appreciated. ************************************************* F. Scott Miller smiller-at-umr.edu Electron Microscope Lab University of Missouri-Rolla voice: 314 341 4727 fax: 314 341 6934 *************************************************
A postdoctoral position is available in TEM characterization of high temperature superconducting (HTSC) materials in the Metals and Ceramics Division of Oak Ridge National Laboratory (ORNL). Job responsibilities would include characterization using advanced EM techniques (EDS, HREM, EELS, CBED, etc.) of a variety of oxide-based HTSC materials. Candidates *must* have a PhD in materials science or related field with a solid background *and* extensive experience in characterization of HTSC materials.
*** DO NOT REPLY ELECTRONICALLY *** *** ELECTRONIC RESPONSES WILL NOT BE CONSIDERED *** *** RESPOND ONLY BY MAIL***
To apply, send a resume, publication list, and names of at least three references to:
G. Sims Oak Ridge National Laboratory P. O. Box 2008 Building 4500S, MS- 6116 Oak Ridge, TN 37831-6116
Oak Ridge National Laboratory is an Equal Opportunity/Affirmative Action Employer.
A postdoctoral position is available in TEM characterization of high temperature superconducting (HTSC) materials in the Metals and Ceramics Division of Oak Ridge National Laboratory (ORNL). Job responsibilities would include characterization using advanced EM techniques (EDS, HREM, EELS, CBED, etc.) of a variety of oxide-based HTSC materials. Candidates *must* have a PhD in materials science or related field with a solid background *and* extensive experience in characterization of HTSC materials.
*** DO NOT REPLY ELECTRONICALLY *** *** ELECTRONIC RESPONSES WILL NOT BE CONSIDERED *** *** RESPOND ONLY BY MAIL***
To apply, send a resume, publication list, and names of at least three references to:
G. Sims Oak Ridge National Laboratory P. O. Box 2008 Building 4500S, MS- 6116 Oak Ridge, TN 37831-6116
Oak Ridge National Laboratory is an Equal Opportunity/Affirmative Action Employer.
I use Polapan400 exclusively, the contrast is as good as Type 52, IMHO, and not needing coating is too good to pass up.
You can get a discount if you order the "bulk pack" - 10 boxes/case (no outer packing)
It does not, however, have a negative with it. If you need a good 4X5 neg, shoot sheet film!
James Long (no relation ;-))
On 24 May 1994, Liang, Long wrote:
} } I was told that there is a new Polaroid product called 'Polapan 400' } which is a medium-speed (ISO 400), coaterless, black-and-white } professional film. The Polaroid company claims that this film is a new } choice for T52 and T53 applications. } } Does anyone have used this new film before? Can I get some comments from } you? I use a SEM and take a lot of T53 films on rocks and minerals. } Thanks. } } } Long Liang } Electron Microprobe and SEM Lab } ARCO Exploration and Production Technology } 2300 West Plano Parkway } Plano, TX 75075 } E-mail : LLIANG-at-is.Arco.COM } } } }
We were just discussing that same problem yesterday with those same scopes. The solution in this department has been to put them in a closet and buy new ones. I found there is a closet full..... and I thought we ought to be able to fix them. I couldn't get the prism straight myself... so if anyone knows of an economical way to fix them, let us know. Thanks ... Nina Allen
Your local microscope repair facility should be able to fix those easily. If not, McBain Instruments in Chatsworth, Calif. certainly can. Their phone # is (818)998-2702.
Following the protocol of Bendayan I complexed Dnas, phospholipase and prteinase_K to colloidal gold but totally failed with RNase A using ph 9.3 and 7.9 and 5.6. I tried to use Sigma Cat. # R 5503 ribonuclease A from bovine pancreas described as being essentially protease and salt free. How ever when I added 0.2ml of a 5mg/ml pure water solution to 10 mls of colloidal gold it immediately turned blue/purple and begain to settle out justas if I had added a high concentration of salt.
Where did I go wrong???? ********************************************************** * Greg Erdos ** * * Director, ICBR EMCL ** Phone 904-392-1295 * * 218 Carr Hall ** FAX 904-392-8598 * * University of Florida ** gwerdos-at-gnv.ifas.ufl.edu * * Gainesville, FL 32611 ** * **********************************************************
I am unable to achieve uniform illumination and focus on my prints, particularly with 35 mm (T-max 100) film. Does anyone know if this is a characteristic of the Beseler CB-7 enlarger or is it something to expect with any enlarger? My 50 mm lens (a Schneider Companon-S) gives a sharp grain (as seen with grain magnifier) in the center but considerably more light and focus degradation at the periphery. The pattern is not a simple circle. The 100 mm lens (Rodenstock Apo-Rodagon) gives better uniformity of illumination and focus but the image is not crisp. The lens apertures are 1-2 stops from wide-open. All exterior lens surfaces are clean. I am using the standard enlarger head with recently cleaned condensers and heat shield. The Kodak polycontrast filters are, unfortunately, below the focusing lens. If the Beseler is inadequate for biological EM lab work using 4 x 5, 3.25 x 4, and 35 mm films, does anyone have recommendations for a better enlarger?
Message-Id: {MAILQUEUE-101.940526141013.256-at-bunyip.ph.rmit.edu.au} To: microscopy-at-anlemc.msd.anl.gov
SEM : viewing polyethylene and other "plastics"
Is it possible to gold coat polyethylene in a standard uncooled gold sputter coater without heating the surface so as to modify it ? We are looking specifically at surface detail and do not wish to have it modified at all. We also wish to look at other probably higher melting point plastics as well. John Millar jjmill-at-rmit.edu.au
Rodney L Kuehn reported problems with evenness of ilumination and focussing on an enlarger. I think that the main problem is that the aperture used in the enlarger lens is simply too large (too small an f number). This results in a small depth of focus. If a larger f number is selected, such as f16 or f22, then the depth of field will increase significantly, and the focussing problem should disappear. The unevenness of the illumination is probably also a problem of aperture size and should also be cured by increasing the f number. I hope this is of use.
Dr Douglas J Arrell Mechanical Performance and Joining Institute for Advanced Materials 1755 ZG Petten Netherlands
Greg Erdos asked about precipitation problems during RNase / colloidal Gold complexing. Our experience suggests that concentrations of 0.5mg of most proteins per 100ml of gold (Frens method) is adequate. If the Sigma RNase is not completely salt free there may still be enough salt in there to aggregate the colloid. Try 0.01ml of the 5mg/ml protein solution per 10ml gold at pH5 - it may work - and still give you adequate label(l)ing.
Jan Coetzee Unit for Electron Microscopy Tel:+27-12-420-2075 University of Pretoria Fax:+27-12-342-1738 Pretoria Internet:janc-at-ccnet.up.ac.za 0002 South Africa
We have used EMS for some time here without any great problems. The bu1 program does take some time to get used to, and building crystals using the space groups can be rather confusing. If the unit cell looks incomplete, try displaying a 2x2x2 block, as the structure may then become clearer.
Alternatively (the method I prefer) use the option which allows you to specify the lattice type (PFIC etc) and then the positions of the atoms in the motif. Thus by specifying F lattice and giving atoms at +- 1/8,1/8,1/8 the silicon crystal can be built very quickly. (I think, from memory, that the option needed is /b)
The bu1 program will provide a file containing all sorts of useful/useless information. In this is a count of the number of atoms in the unit cell. Whichever way you build the crystal, check this file to see that you have the right number.
George Burgess Materials Science & Metallurgy Cambridge CB2 3QZ
We have two versions of EMS, one running on a VAX and one on a PC. In both versions it works fine with silicon in x,y,z=1/8,1/8,1/8. If you look at the .prt file created by bu1, you can check if all atoms are there, and in the right place.
Anna Carlsson National Center for HREM Inorganic Chemistry 2 Chemical Center Lund University P.O. Box 124 S-221 00 Lund Sweden Phone: +46 46 108231 Fax: +46 46 104012 e-mail:Anna.Carlsson-at-oorg2.lth.se
} We were just discussing that same problem yesterday with those same } scopes. The solution in this department has been to put them in a closet } and buy new ones. I found there is a closet full..... and I thought we } ought to be able to fix them. I couldn't get the prism straight } myself... so if anyone knows of an economical way to fix them, let us know. } Thanks ... Nina Allen
If you want, you can contact Ron Roos at Leeds Instruments, 800-444-5333. He is supposed to be some kind of repair modification specialist. Leeds buys, sells, and trades all different kinds of older microscopes. Basic service is ~$60 an hour and parts. On site service is available. Other than liking the service this company provides, I have no connection with Leeds.
************************************************* _______________________ * * | | * * | _____ ______ | * Charles J. Butterick * |__| | | |__| * Electron Microscopy Center * | | * Department of Cell Biology and Anatomy * ______| |______ * Texas Tech University Health Sciences Center * | __ __ | * Lubbock, Texas 79430 * |__| | | |__| * USA * | | * Phone 806 743-2706 voice * | | * 806 743-2707 fax * | | * Email hecub-at-ttacs.ttu.edu * | | * * __| |__ * * |_______| *************************************************
The symptoms that Rodney Kuehn described for his enlarger sound to me exactly like a condenser out of alignment; they do not sound like a depth of focus problem. The Besslers that I am familar with have to be adjusted between 4 x 5 use and 35 mm use. If this is not done correctly, or if the bulb is not well centered, you get problems of focus and illumination that resemble those described by Kuehn. HTH, Tobias Baskin
If there is anyone out there still trying to nurse along a JEOL JEM100B TEM, we have just declared ours surplus. With the exception of a few parts that I have already committed, parts are free to a good home. If it's light, I'll ship it to you; if it's heavy, you need to pay the freight. Julian Smith III Biology Winthrop University Rock Hill, SC 29733 803-323-2111 (Vox) 803-323-2246 (Fax)
I was wondering if anyone has any information on film printers that can be hooked up to a computer (that is also connected to an SEM). Can anyone share their experiences (if any) with the use of these film printers with me and of course how much they cost. We had a refurbished Mirus film printer donated to us awhile back, but it didn't work right after awhile.
I hope that these are appropriate questions for this list. I sent almost the same questions to the NIH-Image list as well, but was not sure if that was the appropriate list either.
Thanks in advance for any information, Peling Melville
-------------------------------------------------------------- Peling Melville peling-at-amnh.org Interdepartmental Laboratories American Museum of Natural History
Lowicryl users, I have heard that HM20 resin is no longer available. Can anyone confirm this and/or offer any explanation as to why it is no longer sold.
Thank you.
Allan Mitchell
per Richard Easingwood, South Campus EM Unit University of Otago, Dunedin New Zealand.
Richard Easingwood Department of Anatomy and Structural Biology, P.O. Box 913 University of Otago, Dunedin, New Zealand. Fax:64-3-479 7254 Telephone:64-3-479 7301
I haven't kept up with this aspect of the field for the last few years, however, many moons ago when I was a younger lad, John Colby (of Magic IV fame) did some work on the shifts in light element K lines. Duncumb et al did work in the mid 60's, Holliday did work on carbide phases around the same time. These shifts are best measured by WDS (but as a uprobist you know this already). The chemical shifts are due to the fact that the emission lines are due to transitions of valence electrons to Kshell vacancies rather than deeper core levels (ie. L's or M's) to the K shell which we see in higher atomic number elements.
My suggestion is to look into some of the more recent books on Bulk X-ray analysis. Love and Scott, or the rewritten book by Reed (which should be on the market soon). You should also probably check out the last few years conference proceedings of the Microbeam Analysis Society.