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?
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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. GšNTHERODT, 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, H“pital 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 M‚decine, Bd Jean Moulin, 13005 Marseille, France. ##Centre de Microscopie ‚lectronique et de Microanalyse, Facult‚ des Sciences et Techniques de St J‚rome, 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, Universita„t 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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------------------ RFC822 Header Follows ------------------ Received: by mse.engin.umich.edu with SMTP;16 Feb 1994 16:48:32 U Received: from localhost by totalrecall.rs.itd.umich.edu (8.6.4/2.2) with X.500 id QAA05927; Wed, 16 Feb 1994 16:48:05 -0500 Received: from anlemc.msd.anl.gov by totalrecall.rs.itd.umich.edu (8.6.4/2.2) with SMTP id QAA05923; Wed, 16 Feb 1994 16:48:03 -0500
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.
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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.