Below is the second circular for next year's Micro conference. Please forward to interested parties or post on your bulletin boards (wood or electronic!). Thanks. John Mansfield (This is a little cleaned version c.f.the first copy and is therefore a little more readable for North American readers. There were too many special characters in the first version - pound signs and the like)
-------------------------------------- MICRO 94
International Microscopy and Image Analysis Conference and Exhibition
12 - 15 September 1994 Earls Court Park Inn, Lillie Road, London
Organized by the Royal Microscopical Society in association with Microscopy and Analysis
Second Circular
Dates
Conference: Monday 12 September - 2.00 pm - 4.15 pm Tuesday 13 September - 10.00 am - 4.15 pm Wednesday 14 September - 10.00 am - 4.15 pm Thursday 15 September - 10.00 am - 4.15 pm
Exhibition: Monday 12 September - 2.00 pm - 7.00 pm Tuesday 13 September - 9.30 am - 6.00 pm Wednesday 14 September - 9.30 am - 6.00 pm Thursday 15 September - 9.30 am - 4.30 pm#012#
On the Monday evening there will be a wine reception at 5.30pm, followed by the AGM and Presidental Lecture at 7.00pm.
CONFERENCE
Scientific Programme The Conference will be run in seven half-day sessions. The Electron Microscopy and Analysis Group of the Institute of Physics (EMAG) and The Physiological Society are each sponsoring separate lectures within the Conference.
The Programme will consist of tutorial lectures and posters and will feature the following topics:-
Monday 12 September (pm) - Materials I EM in the assessment of semiconductor epitaxial growth Reactions to the surface of implanted bioceramics X-Ray microanalysis in biomaterials Optically active nanostructured materials
Tuesday 13 September (am) - Materials II (including EMAG) Auger electron spectroscopy Imaging time of flight SIMS Formation of strained layer superlattices by phase separation Electron microscopy of weakly ordered III-V semiconductor materials
Tuesday 13 September (pm) Scanning Probe Microscopy Applications of atomic force microscopy to thin film research and technology Scanning probe microscopy: near field imaging of surfaces using electrons, forces and photons SPM of living biological systems Environmental SEM
Wednesday 14 September (am) - Image Processing and Analysis Sampling in 3D for quantitative microscopy Digital image processing techniques Image analysis of multicoloured biological specimens In vivo microscopy by video imaging
Wednesday 14 September (pm) - 3D Microscopy Supercomputing in confocal microscopy 3D atomic-scale microanalysis of materials Spatial distribution of fibres in composite materials Confocal polarised-light microscopy
Thursday 15 September (am) - Flow Cytometry and Proliferation Markers Cell cycle control Proliferation-related proteins Proliferation in human tumours Apoptosis
Thursday 15 September (pm) - Living Cell Cytochemistry (including The Physiological Society) Living cell cytochemistry: Ratio Imaging Living cell cytochemistry: Confocal scanning laser microscopy
Thursday 15 September (pm) - Proteases in (patho) physiological processes Use of selective protease inhibitors in the study of collagen breakdown Role of proteases in invasion and metastasis of cancer cells, arthritis and rheumatism and infections
Experts in the fields listed above have been invited to give lectures. Each speaker will provide a review of the particular topic in question and ample time for discussion will be provided.
In addition to the above, a special session will be held on the afternoon of Monday 12 September on how to use the light microscope.
Technical Lectures will be organized by Exhibitors to act as a bridge between the specialized review lectures and the equipment being exhibited.
INVITED SPEAKERS
It is hoped that the following people will be presenting papers at the MICRO 94 Conference:-
Dr Paul D. Brown (University of Cambridge) Professor Peter Marquis (University of Birmingham) Dr Henk Koerten (University of Leiden, The Netherlands) Dr Peter Dobson (University of Oxford) Dr R. K. Wild (University of Bristol) Dr Paul Denison (University of Sheffield) Dr Andrew Norman (Imperial College, London) Dr Caroline Baxter (University of Cambridge) Dr Alan Pidduck (RSRE, Malvern) Dr M. Miles (HH Wills Physics Laboratory, Bristol) Dr H Hoirber(European Molecular Biology Laboratory, Heidelberg, Germany) Mr Chris Gilpin (Manchester Biological EM Centre) Dr Vyvyan Howard (Royal Liverpool Children's Hospital) Dr Dominique Jeulin (Ecole Nationale Superiere des Mines, Fontainebleau, France) Dr Hans Tanke (University of Leiden, The Netherlands) Dr Andreas Kriete (Der Justus Liebig Universitat, Germany) Dr Alfred Cerezo (University of Oxford) Dr A. R. Clarke (University of Leeds) Dr Alan Entwistle (Ludwig Institute for Cancer Research, London) Dr A Bagg (TNO Rijswijk, The Netherlands) Dr Michael Ormerod (Sutton, Surrey) Dr Peter van Mier (Washington University School of Medicine, USA) Professor P. A. McNaughton (King's College London) Dr R. Jacob (King's College London) Dr Vincent Everts (University of Amsterdam, The Netherlands) Dr Ron Van Noorden (University of Amsterdam, The Netherlands)
CONTRIBUTED PAPERS In addition to invited papers, contributions are invited on all aspects of microscopy and related techniques.
All contributed papers will appear in the poster sessions of the Conference. Time will be allowed in the Programme for the viewing of posters, and posters will be on display for the maximum time possible. At certain times authors will be in attendance by their posters to discuss their work.
Camera-ready sheets and instructions for the submission of short abstracts can be obtained from the Royal Microscopical Society office. The deadline for submission is 4 May 1994. These abstracts will appear in the Conference Programme, which will be published in a special MICRO 94 issue of the Proceedings of the Royal Microscopical Society.
Authors will be notified regarding acceptance of their papers by the end of June 1994.
EXHIBITION An Exhibition of the latest microscopes and ancillary instrumentation and equipment will be held at the Earls Court Park Inn, adjacent to the Lecture Theatre. Admission to the Exhibition is free, by conference badge, or by exhibition only badge which will be obtainable at the registration desk.
By 1 November 1993, the following firms had reserved exhibition space:-
Agar Scientific Ltd Alrad Instruments Ltd Bemax (UK) Ltd Bio-Rad Laboratories Ltd British BioCell International Burleigh Instruments (UK) Ltd Cambridge Scanning Co Ltd Confocal Technologies Ltd Cryophysics Ltd Data Cell Ltd Drukker International Edwards High Vacuum International Emitech Ltd Finlay Microvision Co Ltd Fisons Instruments Foster Findlay Associates Ltd Hamamatsu Photonics UK Ltd Hitachi Scientific Instruments ISS Imaging Associates Ltd J K Instruments Ltd JEOL UK Ltd K E Developments Ltd Lasertec Corporation Leica Cambridge Limited Leica UK Limited Microfield Scientific Ltd Microscopy and Analysis Newport Ltd Nikon UK Limited Olympus Optical Co (UK) Ltd Oxford Instruments Microanalysis Group Oxford Instruments Philips Electron Optics Photonic Science Polaroid (UK) Ltd Princeton Gamma-Tech (UK) Ltd Pyser (Holdings) plc Synoptics Ltd Taab Laboratories Equipment Ltd Tracor Europa Carl Zeiss (Oberkochen) Ltd
RECEPTION AND ASSOCIATED EVENTS On Monday 12 September there will be a wine reception in the Exhibition between 5.30 pm and 7.00 pm. The Annual General Meeting of the Royal Microscopical Society, and the Presidential Address will also take place during the evening.
ACCOMMODATION Academic Accommodation A limited amount of academic accommodation has been booked for the use of delegates at Imperial College. Rooms have been booked there on a bed and breakfast basis at a cost of 26 pounds sterling per night. This academic /student accommodation will be filled on a 'first-come first-served' basis. From the nearby Underground Station at South Kensington, Earls Court is two stops along the District or Piccadilly Line.
Hotel Accommodation There are some rooms available in the Earls Court Park Inn at the special MICRO 94 rate of 65.00 pounds per night. If you would like to reserve accommodation at these special rates, please contact the Earls Court Park Inn directly, quoting that you are a MICRO 94 visitor. To be sure of booking rooms at these rates, it is advisable that you book well in advance. Telephone: 071 385 1255 Telex: 917728 Fax: 071 381 4450.
Other Hotel Accommodation Delegates who wish to make their own accommodation arrangements may wish to use the services of Expotel Executive Travel - Europe's leading hotel booking agent, who have been appointed the official hotel agency for MICRO 94. The hotel of your choice or a similar alternative can be booked through Expotel often at discounted rates. By making one telephone call to Expotel on 071 735 0060 stating the event code 'MICRO 94', your reservation will be confirmed verbally followed by confirmation in writing. This free booking service is available to anyone attending MICRO 94. Telephone: 071 735 0060 Telex: 8811951 EXPOTL G Fax: 071 735 2839.
REGISTRATION AND PAYMENT
Registration Registration will take place at the Earls Court Park Inn from 1.00 pm on Monday 12 September 1994, and from 9.00 am on subsequent mornings. Cost of registration for the whole conference is 60 pounds for RMS members and 90 pounds for non-members. Daily rates are 20 pounds for RMS members and 30 pounds for non-members.
Payment Payment may be made by sterling cheque payable to the Royal Microscopical Society (please add 12.00 pounds to cover exchange and bank charges if the cheque is not a UK Bank Cheque) or by credit card (Visa/Barclaycard or Access/Eurocard/Mastercard).
Cancellation and Refunds Cancellations received before 12 July 1994 will be subject to a full refund. No refunds will be made if cancellation is made after this date.
HOW TO GET THERE
The Conference, Exhibition, Posters, and Refreshments will all take place at the Earls Court Park Inn, Lillie Road, London SW6.
MICRO 94 is organized by the Royal Microscopical Society, 37/38 St Clements, Oxford OX4 1AJ, UK, in association with Microscopy and Analysis. Telephone: 0865 248768, Fax: 0865 791237, Email: RMS-at-UK.AC.OX.VAX.
I've been involved in design of an EM lab, a histology lab and most recently given freehand to design a facility with "ultimate, within the budget" two general histology labs, a microtome room and two microscope/image analysis rooms to be built soon.
Think carefully about work flow and separation of incompatible activities. Traffic patterns can be aggravating if their is a choke point. e.g. our peninsula bench has ahighly used sink on the end and only 3.5 ft to the wall. There is a blackboard and garbage cans along that wall, with lots of traffic through this narrow slot. Don't allow the free edge of open doors to aim toward knees or heads on single doored cupboards. Big, deep double sinks. Double sinks, or many sinks, ensures that processes requiring long washes don't tie up the sink, and minimize splatter. Our PI put in a lot of pocket sinks in our existing lab. they are great if the benches are not cluttered. Otherwise they will fill with pencils, towels and BEEM capsules as well as splatter adjacent equipment. We have 4-12 people woking in 400 sq. ft. We are vey cluttered and the pocket sinks are useless.
Dont put fume hoods in corners. That makes the wall-end foot or so harder to use, especially if you end up storing anything on the floor in that corner. Stuff on the floor also absturcts the cupboard doors below the hood.
It took some doing, but I got approval for an awning hood over one bench in our new histology lab. It is 10 ft. long, wasn't enough room to go longer. All embedding, deparaffinizing, staining and coverslipping will take place under it. An open shelf below will hold vacuum ovens for paraffin and the plastics oven so they can vent upward from the back. Osmium and fix preparation will remain in the standard hood (a 6 ft. hood). Drafts from ventilation have been a major pain for us and have required makeshift deflectors on vents, or finding combinations of closed doors and hood sashes to slow down drafts.
Consider separate 110V circuits for computers and everything else, especially if you have any surge producers like pump motors or uv lamps.
Look out for dust collectors.. They are pricey, but put glass doored cupboard over dust sensitive situations like microtomes and grid staining area. These will not present so many dusty surfaces as will open shelving.
Gotta go, but one last issue, physically touch, test, open and close the furniture before allowing it to go in. some looks great but is worse than useless.
Prior comments about wiring and fields are very true. We are seeking a new home for our SEM because of fields. (Keep an eye on remodelling - I remember a Philips 300 whose beam would deflect everytime the new photocopier next door made a copy)
On Tue, 30 Nov 1993, Greg Erdos ICBR EM Core Lab University of Florida wrote:
} I am now in the process of designing a new EM suite dedicated to biological } EM. I would like input from those of you who work in what they feel is an } ideal or close to ideal physical arrangement. } I would also like to hear from those who have envisioned an ideal } EM suite but have never seen it built. The restrictions are that a space of } 2200 sq ft should house 2 TEMs and 2 SEMs as well as a prep lab, darkrooms } ancillary equipment and 2 offices. The overall design is of more interest } than actual use of the stated space.
Greg, we are well into the final design of our EM facility for the new Cell and Mol. Bio building in the Div. of Bio Sci. I based our design on the existing EM suite which I remodeled about 10 yrs ago. I also used the book by R. H. Alderson, "Design of the Electron Microscope Laboratory", part of Audrey Glauert's series in Practical Methods in Electron Microscopy published by North-Holland/American Elsevier ISBN 0 444 10816 5. I can send you plans although I doubt if the architects would like that. Further, my design was limited to the space available in the new building and the shape of the room modules. Each module is about 10 ft by 30 ft. I used 3 modules for the main lab and office but there is extra space available across the hall where I housed the freeze fracture device, the microtomes, student TEM, and dept. darkrooms (also under my supervision). You can reach me at 916 752 2914, Univ. of Cal. at Davis, Dept. of Evolution and Ecology. Rick A. Harris
Hitachi H-600 STEM: 13 years old, under service contract with Hitachi until this year, in most excellant condition, scope has all service reports and manuals.
Original cost: $175,000 Asking price: Make offer I can't refuse Contact: Phil Rutledge, Director, Center for Electron Microscopy University of Maryland Baltimore County Dept. of Biology Catonsville, MD 21228 USA Phone: (410) 455-3582 FAX: (410) 455-3875 Email: prutle1-at-umbc.edu
I am looking for a supplier of a 2D array detector (for luminescence microscopy measurements) which will operate out to 1.6 microns. Essentially the same thing as a Si CCD except made from a smaller gap material (InGaAs?, Ge?). I would like recommendations of a manufacturer/supplier.
Erik Reuter, e-reuter-at-uiuc.edu, 217-244-6378 phone, 217-244-6375 fax
Here is a list of references I received from my inquiry on light microscopy. Thanks to everyone who contributed!
***
The best set of notes on LM theory I know of are the course notes for the AQLM course run at Woods Hole each year, they are long and complex but in my experience a great resource
Kodak publications: "Photography Through The Microscope", P-2, cat.# 152-8371 "Kodak Scientific Imaging Products", L-10, cat. # 813-9321 The first has loads of basic information about objectives, condensers, setting-up illumination, flourescence, phase contrast, etc. Both have bibliographies.
Title : Optical Microscopy : Emerging Methods and Applications Author : Herman, Brian/Lemasters, John J. (Editors) ISBN : 0123420601 Subject : Non-Fiction Dewey # : 578.00 Publisher: Academic Pr Date Pub : 12/92 Binding : Hardcover+ Price : $ 79.95
Loveland, Roger Platt. Photomicrography; a comprehensive treatise [by] Roger P. Loveland. New York, Wiley [1970] 2 v. (x, 1039, I-xii p.) illus. 23 cm. LC CALL NUMBER: QH251 .L68 SUBJECTS: Photomicrography. SERIES TITLES (Indexed under SERI option): Wiley series on photographic science and technology and the graphic arts DEWEY DEC: 778.3/11 NOTES: Includes bibliographies. ISBN: 0471548308 LCCN: 70-88315 r892
Shillaber, Charles Patten, 1886- Photomicrography in theory and practice, New York, J. Wiley & sons, inc.; London, Chapman & Hall, limited [1944] viii, 773 p. incl. illus., tables, diagrs. 22 cm. LC CALL NUMBER: QH251 .S5 SUBJECTS: Photomicrography. LCCN: 44-6507
Light microscopy in biology : a practical approach / edited by Alan J. Lacey. Oxford, England ; New York : IRL Press, c1989. xviii, 329 p. : ill. ; 23 cm. LC CALL NUMBER: QH207 .L49 1989 SUBJECTS: Microscopy--Technique. ADDED ENTRIES: Lacey, Alan J. SERIES TITLES (Indexed under SERI option): Practical approach series DEWEY DEC: 578/.4 dc19 NOTES: Includes bibliographies and index. ISBN: 0199630364 : $54.00 (U.S. : est.) 0199630372 (soft) : $36.00 (U.S. : est.) LCCN: 88-23505 r92
Pluta, Maksymilian. Advanced light microscopy / Maksymilian Pluta. Warszawa : PWN ; Amsterdam ; New York : Elsevier : Distribution for the USA and Canada, Elsevier Science P ublishing Co., 1988- {1989 } v. 1- {2 } : ill. ; 25 cm. LC CALL NUMBER: QH207 .P54 1988 SUBJECTS: Microscopy--Technique. DEWEY DEC: 502/.8/2 dc19 CONTENTS (Incomplete): v. 1. Principles and basic properties -- v. 2. Speciali zed methods. NOTES: Translated from the Polish manuscript. Includes bibliographical references and index. ISBN: 0444989390 (v. 1) : $175.00 (est.) 0444989188 (v. 2) LCCN: 87-24605 r922
Ploem, J. S. Introduction to fluorescence microscopy / J.S. Ploem and H.J. Tanke. Oxford ; New York : Oxford University Press ; Oxford : Royal Microscopical Society, 1987. vi, 56 p. : ill. ; 24 cm. LC CALL NUMBER: QH212.F55 P57 1987 SUBJECTS: Fluorescence microscopy. ADDED ENTRIES: Tanke, H. J. SERIES TITLES (Indexed under SERI option): Oxford science publications Microscopy handbooks / Royal Microscopical Society ; 10 Microscopy handbooks ; 10. DEWEY DEC: 578/.4 dc19 NOTES: Includes bibliographies and index. ISBN: 0198564082 : $7.50 (U.S.) LCCN: 87-5539
Inoue, Shinya. Video microscopy / Shinya Inoue ; with contributions by Robert J. Walker, Jr. ... [et al.]. New York : Plenum Press, c1986. xxvi, 584 p. : ill. (some col.) ; 26 cm. LC CALL NUMBER: QH222 .I56 1986 SUBJECTS: Video microscopy. DEWEY DEC: 578/.4 dc19 NOTES: Includes index. Bibliography: p. 515-529. ISBN: 0306421208 LCCN: 85-28252
Slayter, Elizabeth M. Light and electron microscopy / Elizabeth M. Slayter, Henry S. Slayter. Cambridge [England] ; New York : Cambridge University Press, 1993. p. cm. LC CALL NUMBER: QH205.2 .S54 1993 SUBJECTS: Microscopy. Compound microscopes. Electron microscopy. ADDED ENTRIES: Slayter, Henry S. DEWEY DEC: 578/.4 dc20 NOTES: Includes indexes. 92-7825 (continued): ISBN: 0521327148 LCCN: 92-7825 r93
Russ, John C. The image processing handbook / John C. Russ. Boca Raton, Fla. : CRC Press, c1992. 445 p. : ill. (some col.) ; 27 cm. LC CALL NUMBER: TA1632 .R88 1992 SUBJECTS: Image processing. ADDED ENTRIES: Image processing. DEWEY DEC: 621.36/7 dc20 NOTES: Includes bibliographical references (p. [435]-442) and index. ISBN: 0849342333 (acid-free) LCCN: 92-4936
Russ, John C. Computer-assisted microscopy : the measurement and analysis of images / John C. Russ. New York : Plenum Press, c1990. xii, 453 p. : ill. ; 26 cm. LC CALL NUMBER: TA1632 .R87 1990 SUBJECTS: Image processing. Microscopy--Data processing. Optical pattern recognition. DEWEY DEC: 502/.8/20285 dc20 NOTES: Includes bibliographical references and index. ISBN: 0306434105 LCCN: 89-70945 r92
Bradbury, Savile. Basic measurement techniques for light microscopy / SavileBradbury. Oxford ; New York : Oxford University Press ; [London] : Royal Microscopial Society, c1991. viii, 97 p. : ill. ; 24 cm. LC CALL NUMBER: QH207 .B74 1991 SUBJECTS: Microscopy--Measurement. SERIES TITLES (Indexed under SERI option): Royal Microscopical Society microscopy handbooks ; 23 Microscopy handbooks ; 23. DEWEY DEC: 502/.8/2 dc20 NOTES: Includes bibliographical references and index. ISBN: 0198564260 : $14.90 LCCN: 90-21567 r92
Bradbury, Savile. An introduction to the optical microscope / Savile Bradbury. Rev. ed. Oxford ; New York : Oxford University Press ; Oxford : Royal Microscopical Society, c1988. 86 p. : ill. ; 24 cm. LC CALL NUMBER: QH205.2 .B67 1988 SUBJECTS: Microscopes. Microscopy. SERIES TITLES (Indexed under SERI option): Microscopy handbooks ; 1 Oxford science publications DEWEY DEC: 535/.332 dc19 NOTES: Includes bibliographical references (p. [82]). ISBN: 0198564198 : $6.00 LCCN: 88-38921 r92
Dictionary of light microscopy / compiled by the Nomenclature Committee of the RMS, S. Bradbury ... [et al.]. Oxford ; New York : Oxford University Press ; Oxford : Royal Microscopical Society, 1989. x, 139 p. : ill. ; 25 cm. LC CALL NUMBER: QH203 .D53 1989 SUBJECTS: Microscopy--Dictionaries. ADDED ENTRIES: Bradbury, Savile. Royal Microscopical Society (Great Britain). Nomenclature Committee. RMS dictionary of light microscopy. SERIES TITLES (Indexed under SERI option): Microscopy handbooks ; 15 Oxford science publications DEWEY DEC: 502/.8/2 dc19 NOTES: Cover title: RMS dictionary of light microscopy. ISBN: 019856421X : $32.00 (U.S.) 0198564139 (pbk.) : $14.95 (U.S.) LCCN: 88-22712 r92
Thomson, D. J. An introduction to photomicrography / D.J. Thomson, Savile Bradbury. Oxford [Oxfordshire] ; New York : Oxford University Press ; Oxford : Royal Microscopical Society, 1987. 74 p. : ill. (some col.) ; 24 cm. LC CALL NUMBER: QH251 .T44 1987 SUBJECTS: Photomicrography. ADDED ENTRIES: Bradbury, Savile. SERIES TITLES (Indexed under SERI option): Microscopy handbooks ; 13 DEWEY DEC: 578/.4 dc19 NOTES: Includes index. Bibliography: p. [69]-70. ISBN: 0198564147 (U.S. : pbk.) : $9.00 LCCN: 86-33220
Bradbury, Savile. The evolution of the microscope, by S. Bradbury. [1st ed.]. Oxford, New York, Pergamon Press [1967] x, 357 p. illus. 22 cm. LC CALL NUMBER: QH204 .B7 SUBJECTS: Microscopes--History. DEWEY DEC: 578/.09 NOTES: Bibliography: p. 347-348. LCCN: 67-18485 r922
Erik Reuter, e-reuter-at-uiuc.edu, 217-244-6378 phone, 217-244-6375 fax
This message was posted on a different forum however it has applicabity here too. I've reflected it for the subscribers:
Nestor Z. ANL EMCenter ====================
To: Multiple recipients of list {nih-image-at-soils.umn.edu}
} I am interested in finding references to algorithms for grain size } measurement. Any help would be welcome. } There are two "standard" methods for measuring the property usually called "grain size." One is based on counting the number of grains visible per unit area on a two-d section through the structure, and the other is based on drawing lines across the same image and coutning the number of intersections of the lines with grain boundaries. Neither actually measues the "size" of the grains, which are 3D entities. From the point of view of stereology, the second method measures the amount of grain boundary area per unit volume in the sample, which is often important to properties (mechanical, electrical, etc.) because many things happen at grain boundaries. The first method measures the total length per unit volume of the "triple line" where three grains meet in the volume. This is particularly important in sintered structures since the pore channels and much diffusion occurs there. But NEITHER is the "grain size." It happens that in many materials (but by no means all), the grain size distribution after standardized heat treatments (e.g., full recrystallization) is sufficiently consistent that there is a more-or-less consistent relationship between the actual size distibution of the grains and these other parameters. This combined with the fact that people (or at least metallurgists) have been measuring these properties for most of this century, and CALLING it grain size, has made these measurements an accepted standard tool for microstructural characterization.
There are a couple of fairly robust computer approaches to measuring each of the two different properties. In the computer case, instead of drawing lines and couting crossing points it is easier to first perform whatever image processing is needed to threshold and skeletonize the grain boundaries and then measure the total length (properly, not just counting pixels but taking into account the geometry of the line). This is simply related to the intercept count and the surface are per unit volume, and hence to the "grain size number." It has the disadvantage that many grain images do not show all of the boundaries well (inadequate etching or polishing, ec.) The algorithm that works best for the other method is to count the triple points in the grain boundary image, which usually do show up well and are easily define as points on the skeleton that have more than 2 neighbors. This gives the total length of triple line, and again a "grain size number."
For details on both algorithms, and the equations to convert the values, see p.147 and p. 225 in Computer Assisted Microscopy, J. C. Russ, 1990, Plenum Press. ISBN 0-306-43410-5
There are other issues to consider as well, including processing operations on grain images that may alter the length of boundaries (watershed segmentation, for instance, is either a boon or a curse), and ways to actually measure the 3D distribution of grain sizes, without resorting to very biased assumptions like treating the grains as spheres (!). The field of stereology has dealt with such problems for decades, and there are actually some pretty good answers to most of them. Good, but not necessarily simple to implement.
__________________________________________________ John Russ (John_Russ-at-ncsu.edu) or (russ-at-mat.mte.ncsu.edu) Materials Science and Engineering Department, North Carolina State Univ., Raleigh, NC 27695-7907 phone: 919-515-3328 fax: 919-515-7724
On Fri, 3 Dec 1993 03:14:37 +0200, Steve Barlow wrote:
} } We are considering the purchase of a new diamond knife and were interested } to know if anyone has had any experience with companies other than } Diatome. In specific, Edgecraft Corp, MicroStar, or DiaTech. } } } From personal experience, I know that Diatome makes an excellent knife. } Dupont knives were good, but I haven't used any of their knives since they } were bought out by DDK. DiaTech has been touting its 'unique' mounting } system that greatly reduces re-sharpening costs. } } If anyone can offer their own experiences, I would appreciate it, } particularly regarding the quality of the resharpening. } } ---------------------------------------------------------- } Dr. Steven Barlow } EM Facility/Biology Dept. } San Diego State University } San Diego CA 92182-0057 } phone: (619) 594-4523 } fax: (619) 594-5676 } email to sbarlow-at-sunstroke.sdsu.edu
Greetings from the "old continent" to all microscopy-netters,
We have used Diatome knifes and found them excellent. Now we are going to test-section a Drukker-knife from Drukker International B.V., the Netherlands. I have great expectations for this particular maker, since they claim to have made the diamond surface hydrophilic, thus enabling sectioning with not-filled waterlevel. We will know more about this knife in two months.
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I have been doing E.M. for 25 years and for the past 10-12years have been using MicroStar knives. The knives are of excellant quality. I have never had a problem with any of their knives. I have used DuPont and Diatome but as long as MicroStar remains in business, I will continue to use their knives. Their people are friendly, informative and very helpful. As a whole, the company is excellant to deal with. I recommend their knives whole heartedly. Questions? Phil Rutledge, Director, Center for E.M. UMBC Dept. of Biology Catonsville, MD 21228 USA Phone:(410)455-3582 FAX: (410)455-3875 Email: prutle1-at-gl.umbc.edu
I have had excellent service from Microstar for many years, Both with new knives and with resharpening knives that originally came from several different vendors. I'd say we have gotten about 20 knives from this company and have always been satisfied. This is not to say that other suppliers couldn't do as well. My recent (10 yrs) experience has only been with Microstar.
From : JOSEPH MCGINN Number : 8 of 9 To : ALL Date : 12/02/93 1:08a Subject : Nicalon-Black Glass Reference : NONE Read : [N/A] Private : NO Conf : 005 - Specimen Preparation
Has anyone had experience with sample preparation of this composite system. There is a real problem retaining the fibers in the matrix during the final stages of mechanical polishing. Has anyone found a trick to get these samples below 20 microns for ion milling?
From : CRAIG LENDING Number : 9 of 9 To : ALL Date : 12/03/93 10:54a Subject : Unicryl embedding Reference : NONE Read : [N/A] Private : NO Conf : 005 - Specimen Preparation
Has anyone had any experience with immunolabelling with the new Unicryl resin that is marketed by SPI? I currently use LR White, but they claim that this resin has better stability under the electron beam and also gives higher levels of labelling than LR White -- at least that's what they say!
I have tried both resins and have had good results with both. The only problem I have had with LR White is getting my students or principle investigators remembering to use 80kv. LR White is best used at voltages above 60kv or you can have resin "droppings" formed on the wall of the projector pole piece. I have 4 E.M.s and since I don't have service contracts I end up cleaning the contaminated scope. If you can use voltages above 60kv, you can get equally good results with either resin. I normally use 80kv on all of my scopes at all times and have never tried Unicryl at 60kv. It may be OK to use at 60kv or you may have to use a higher voltage as I have to do with LR White. The only real difference I have seen is the initial cost of the two resins. Good Luck! Phone: (410)455-3582 FAX: (410)455-3875 Email: prutle1-at-gl.umbc.edu
We are contemplating getting a new ultra-high resolution field emission SEM. This would be for applications in both biological and materials fields. Resolution needs to be better than 1nm at ca 15kV. We are at present operating 2x SEM's and 1 TEM in a service unit, so this new SEM would be dedicated to clean samples for hi-res work. I would appreciate any and all input from users regarding their experience of such equipment (microscopes and necessary prep peripherals). Practical experience of cold versus thermal FEG's (stability, lifetime, running costs, vacuum problems etc) especially welcome. Dr J Coetzee Unit for Electron Microscopy Tel:+27-12-420-2075 University of Pretoria Fax:+27-12-342-1738 Pretoria 0002 Internet:janc-at-ccnet.up.ac.za South Africa
On Fri, 3 Dec 1993, Nestor J. Zaluzec (708)-252-5075, -4964 wrote:
} } } } From : CRAIG LENDING Number : 9 of 9 } To : ALL Date : 12/03/93 10:54a } Subject : Unicryl embedding Reference : NONE } Read : [N/A] Private : NO } Conf : 005 - Specimen Preparation } } Has anyone had any experience with immunolabelling with the new Unicryl } resin that is marketed by SPI? I currently use LR White, but they claim } that this resin has better stability under the electron beam and also } gives higher levels of labelling than LR White -- at least that's what } they say! } } ================ } We use unicryl on a regular basis as our main embedding medium. The immunolabelling characteristics are comparable or better than Lowicryl K4M. The morphology (?) even seems to be improved using the same preparation procedures. The cutting characteristics are very well. We cut it using a diatome knife and also using glassknifes. The person in charge of cutting has a lot of experience with different materials (incl LR White) and she thinks Unicryl cuts at least as good as Epon or Araldite. Hoping to hear from you. We are interested in LR White because of the not complete dehydration. We have tried some LR White but have experienced great difficulties. Blocks tend to break and are difficult to cut. Perhaps you could comment.
We've had experience with DDK over many years with many knives. Their knife quality is excellent. The resharpening is excellent. An easy company to deal with.
On Thu, 2 Dec 1993, Steve Barlow wrote:
} We are considering the purchase of a new diamond knife and were interested } to know if anyone has had any experience with companies other than } Diatome. In specific, Edgecraft Corp, MicroStar, or DiaTech. } } } From personal experience, I know that Diatome makes an excellent knife. } Dupont knives were good, but I haven't used any of their knives since they } were bought out by DDK. DiaTech has been touting its 'unique' mounting } system that greatly reduces re-sharpening costs. } } If anyone can offer their own experiences, I would appreciate it, } particularly regarding the quality of the resharpening. } } ---------------------------------------------------------- } Dr. Steven Barlow } EM Facility/Biology Dept. } San Diego State University } San Diego CA 92182-0057 } phone: (619) 594-4523 } fax: (619) 594-5676 } email to sbarlow-at-sunstroke.sdsu.edu }
-------------------------------------- Jim Michael
------------------ RFC822 Header Follows ------------------ Received: by quickmail.yale.edu with SMTP;6 Dec 1993 12:13:44 -0500
At Argonne National Laboratory's E. M. Center, we have used a Kodak Royalprint Processor for ten years or so. It will handle any developer- incorpoorated RC paper and give a dry, archival print in about 45 seconds, as Ron Anderson noted. It is not cheaper than using trays, but you'll be able to do more work in less time with less frustration. Any similar processor will give you the same results. If you want to get really efficient, you can get an automatic dodging enlarger from LogEtronics. Then you don't have to spend time or test strips trying to find the correct exposure! I haven't heard of anyone who is using the 3M Dry Silver Processor which Ted Pella, Inc. is marketing. According to it's brochure, the processor is supposed to deliver a dry, archival print in 6 seconds. You'd avoid handling chemicals with that processor. Any experience out there?
} I am in the process of deciding whether or not to purchase a B&W photo paper } processor for our EM Lab darkroom. I have reviewed some product literature and } now would like to hear from anyone who has used such processors. Are they } neater, cleaner, faster and more economical than tray processing? Are they } 'plumber's nightmares'? Are they tied to a specific paper type and chemistry? }
Ted Pella, Inc. has advertised a product called `Pro Automatic Print and Film Processor' in its recent catalog. It is said that this can process negatives (4489 TM, T-MAx etc.) as well as prints using the same developer in one unit. It costs about $4000. There is yet an another product advertised called the Pelco Dry Silver Processor which does not use any chemicals. The product is marked about $1400. We are considering upgrading our darkroom facilities, and so contemplating on purchasing the Print and Film Processor. Does anyone have any experience with it? Thanx in advance for suggestions/advice.
} } Thank you for any information you can provide. } } Gib G. Ahlstrand } Electron Optical Facility } University of Minnesota
Ananda S. Murthy Department of Physics & Astronomy University of Delaware Newark, DE 19716 (302) 831-6811
The mentions of wash by others reminded me: the system I used still required that the photos be fixed as usual for tray developing. (Using Kodak Polycontrast RC paper.) I forget if it was really required, but yellowing etc. wash a problem if photos weren't fixed. Wash was 5-10 min. Phil Oshel
Re} EM practical courses Back by popular demand -even though we never seem to make a profit (or break even).
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 detailson both courses;
Paul Webster, Department of Cell Biology, Yale School of Medicine, 333 Cedar Street, New Haven, CT 06510.
We too use Ilford 2150RC units and have no problems with yellowing etc. I should note, however, that we have had a lot of minor problems with one of the two identical units in our building and this has resulted in considerable downtime and frustration. When they work properly, they are wonderful units which increase the work rate by at least a factor of 2-3 over manual developing. If, however, I was based in the states and looking to buy a processor I'd be asking my Ilford representative some very probing questions about service and call-out arrangements in case of problems. Please note that our other 2150RC has given about four years of trouble free service.
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 worked part time for two years processing b&w EMs and had extensive experience w/ b&w photography before the job (including making silk screens, photo lithographs, etc.).
Having a Ektagraphic(?) machine by Kodak greatly sped up making large numbers of notebook prints. The machine developed and stabilized in 20-30 seconds or so. I just piled the prints up and fixed and washed them in a big batch before lunch and before going home. I only had to get my hands wet two or three times each day; the actual printing was almost a dry process.
Granted these prints are not archival, but they last for at least five years without significant fading or discoloring.
Another trick for tray processing is to keep dektol (or your favorite developer) warm ( } 70 degrees F); the paper develops instantly, but the chemicals oxidize fast.
nih-image-request-at-nx1.soils.umn.edu (NIH Image Mailing List Maintainer) Newsgroups: news.announce.newgroups,news.groups,sci.chem,sci.engr.chem,sci.geo.geology,sci.materials,sci.misc,sci.physics,sci.optics,sci.research,sci.physics.accelerators,sci.engr,sci.polymers
FIRST CALL FOR VOTES (of 2)
Unmoderated group sci.techniques.microscopy
Newsgroups line:
sci.techniques.microscopy The field of microscopy.
Votes must be received by 23:59:59 UTC, 2 January 1994.
After this CFV appears on news.announce.newgroups it will be sent to the following mailing 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}
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This vote is being conducted by a neutral third party. For voting questions only, contact pschleck-at-unomaha.edu. For questions about the proposed group, contact John F. Mansfield {John.F.Mansfield-at-umich.edu} .
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.
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Standard Guidelines for voting apply. One vote per person, no more than one vote per account. 100 more YES votes than NO votes and twice as many YES votes as NO votes are the requirements for group creation. -- Paul W. Schleck pschleck-at-unomaha.edu
The Journal of Microscopy has received the following books for review:
Enzyme Histochemistry. A Laboratory Manual of Current Methods By C J F Van Noorden & W M Frederiks. Oxford University Press
X-ray Microanalysis in Biology. Experimental Techniques and Applications Edited By David C Sigee, A John Morgan, Adrian T Sumner & Alice Warley. Cambridge University Press
Immunocytochemistry II Edited by A C Cuello. John Wiley
A Manual of Applied Techniques for Biological Electron Microscopy By Michael J Dykstra. Plenum Publishing
Soil Microscopy and Micromorphology By E A Fitzpatrick. John Wiley
If you are interested in reviewing any of these publications, and can provide a review within two months of receiving the book, please contact Gillian Wilson at RMS-at-uk.ac.ox.vax directly. There is no financial reward, but you can keep the book, of course! I will work on a first-come, first-served basis.
I am also looking for volunteers to review a forthcoming book (which will appear in print in January 1994):
In Situ Hybridization By A R Leitch, T Scwarzacher, D Jackson & I J Leitch. BIOS Publications
Two copies will be available for review. ******************************************************************************
RE} Lowicryl protocol In reply to the request for a Lowicryl protocol that works, from Rajesh Patel. Try this for something a little more exotic.
Fix the lung tissue as well as possible (perfusion fix if possible) in either formaldehyde or glutaraldehyde, 1 -2 hrs is enough. Infiltrate with sucrose (2.3M in PBS) for approx. 1 hr, depending on the size of the pieces. Freeze the cryoprotected pieces on the tips of wire stubs (or cryo ultramicrotome specimen stubs if available) by immersion in liquid nitrogen. Do not let them warm up but transfer them directly to 100% methanol on dry ice (precooled of course). Leave until they fall of the wire (usually overnight is enough for small pieces). Wash with fresh, cold methanol on dry ice then gradually replace the methanol with methanol containing increasing amounts of Lowicryl (we use HM-20 or HM-23 at these depressed temperatures) until you can put the tissue in 100% resin. Leave overnight then embed in fresh resin, still on dry ice, and polymerize, on dry ice, using a UV light as per instructions. Remember that oxygen will interfere with the resin polymerization so use embedding capsules or centrifuge tubes that are filled up with resin and are sealed. Also bubble nitrogen through the resin before use.
This easy freeze substitution protocol can be done in a styrofoam box (which is what we use) or in a Revcco type freezer (the Lowicryl smells bad and is difficult to stop from spilling though!) The cold methanol does not need to be specially dehydrated before use (just open a new bottle) and can contain 1% osmium tetroxide or 1% uranyl acetate to improve contrast. Do not allow the tissues to warm up in the osmium if you are concerned about antigenicity. Surprizingly, the additions to the methanol do not seem to affect antigenicity on most of our systems. For extra polymerization Lowicryl blocks can be put under UV light at room temp (or in the sun - says Heinz Schwarz of Tubingen). A reference for this can be found in van Genderen et al 1991 J. Cell Biol. 115:1009-1019.
If your antibodies do not work on Lowicryl-embedded tissue (and many do not) why not try cryosections. They are not difficult to obtain - even from lung. Good luck.
At Image Analysis we have a Tektronix Phaser IISDX Dye Sublimation printer which is connected simultaneously to a Silicon Graphics Indigo (via a parellel port) and a Macintosh (via appletalk). This printer produces near photographic quality (some people consider it better than photo quality) 8.5 x 11 and legal size 24 bit color prints and transparencies. Used in combination with Adobe Photoshop on the Mac or Showcase on the Silicon Graphics, it produces fantastic results allowing researchers to create hardcopy for posters, publications and portfolios in record time. The printer understands most postscript so that text and object oriented graphics is reproduced with little or no aliasing.
The main drawback with this printer is cost per print which is around $3. The newer printers from tektronix and other vendors are rumored to use regular printer paper rather than requiring the purchase of special paper for the printer. Two other problems with the printer have been inability to read all postscript files (usually solved by reading the files into Photoshop or Showcase first) and matching the intensity of the colors on the computer screen with what is produced by the printer.
boyd
Boyd Knosp 319-335-6715 University of Iowa knosp-at-tessa.iaf.uiowa.edu Image Analysis 77 EMRB Iowa City, IA 52242
On Thu, 9 Dec 1993, L. D. Marks wrote:
} } Any comments on computer image printers ? } } Laurie Marks } Northwestern
I am interested in any problems that other users of the Reichert microtome Ultracut S with FCS cryo attachment have had. I am especially interested in chronic problems with the cryo pump; could you also relay what your solutions have been? I am having the membrane in my pump replaced more often than seems appropriate for a new piece of equipment and wonder if this is typical. Also the poly tube that connects the pump to the liquid nitrogen in the dewar keeps slipping off of its connector - anyone having this problem? Solutions? Thanks.
R} Beef Wenbo Zhang writes that he has pepper on his beef muscle after fixation. The muscle is fixed and postfixed in the presence of phosphate buffer which may be the problem. All the glutaraldehyde must be washed out of the tissue before going into osmium otherwise some sort of reaction occurs to cause a precipitate. This may vary from small particles (pepper) to crystals.
Message-Id: {9312101416.AA12900-at-umail.UMD.EDU} To: microscopy-at-anlemc.msd.anl.gov
Fixation pepper probably results from the interaction of a number of key fixation ingredients including phosphate buffer, glutaraldehyde, uranyl acetate and ethanol. For a nice work on the role of the above participants in the formation of dense deposits see the paper by J. Louw, et al in STAIN TECHNOLOGY 65(5): 243-250, 1990 entitled "Electron dense artefactual deposits in tissue sections: The role of ethanol, uranyl acetate and phosphate buffer."
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
Reply_ RE} Printers Reply from: John Mansfield North Campus Electron Microbeam Analysis Laboratory University of Michigan 2455 Hayward Ann Arbor MI 48109-2143 Phone: (313)936-3352 jfmjfm-at-engin.umich.edu or John_Mansfield-at-mse.engin.umich.edu OK, we recently (actually Ron Gibala and John Halloran paid for it) bought a Kodak ColorEase 300 PS printer for printing out images. We are talking about a Mac environment here with a few PCs so this discussion will be centered on printing from Macs for the most part. We had a demo of the ColorEase and it during that the printer was not printing as well as the Tektronix Phaser IIsdi. So, we were going to go with the Tektronix (even though it was more expensive). But during the demo, I started using the Apple Laserwriter 8.0 drivers and then the Kodak performed better for grayscale images. We have found for 8bit color and grey images we use the Apple drivers (we have a PPD downloaded from Adobe.com so the applications on the Macs know big the print area is on the Kodak under 8.0. The 8.0 drivers were developed by Apple and Adobe together anyway). There are PPDs at Adobe for most Postscript printers (except maybe the Hewlett Packards). To get them check out ftp.adobe.com. Anyway after that small diversion, we use the ColorEase driver if we want to do 24bit color and try and match what the image looks like on the screen. Since we dont often use true 24bit color we mostly use the Apple drivers. The latest version of NIH-Image when used in conjunction with the Apple drivers will scale the image you are trying to print to fit on the page and so that avoids the clipping warnings you get from Photoshop. Of course, if you want to use 24bit color you need to use Photoshop. It's a nice printer. The paper and transparencies are a lot more heavy weight that the Tek media. Prints cost us about $2 each and transparencies a little more. Considering you can get 3 Polaroid size prints on one sheet that is a considerable cost saving over Polaroid. The Polaroid that we use is in excess of $2.50 per shot and so if you use the Kodak printer, you can pay for the printer after 3500 prints. Given the atrocious Polaroid quality and their vague responses if you try and get credit for a bad batch of film, this printer seems to me like an excellent way to go if you have a good way of electronically acquiring your images. As a disclaimer, my opinions of the corporations that I mention above are my own personal ones and do not in any way reflect the opinions of my employer
We have a Codonics NP-600 dye sub printer. It came to us as part of a confocal package, the price is about $10K. Cost of the standard B&W/color paper is $1.50 a sheet (8" x 8.5"). The 8.5" x 11 is about $3 a print. Microscopy images are as good as one's equipment. Color matching seems good but we may not be as discriminating as others. One major strongpoint is the number of acceptable formats : IFF, GIF, JPEG, PICT, PCX, PPM, RGB, Sun Raster, TIFF, TGA, X-11 Window Dump, CMU WM, IMG, PBM, PGM, and Lisp Machine Bitmap. Codonics is a small outfit out of Ohio and have provided excellent backup. Take a look. ************************************************* * * * * * 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 * * * * * *************************************************
We have a vast amount of experience with the processing of myocardial / muscular tissue. In our experience it is the phosphates found in muscular tissue which play a role in the formation of these dense deposits.
The essential factors in the formation of electron dense deposits appear to be phosphate buffer, ethanol and uranyl acetate. Glutaraldehyde may contribute in a way, while osmium does not appear to play a role. (See our article in STAIN TECHNOLOGY 65: 243-250 (1990). -------
Message-ID: {MAILQUEUE-101.931213111417.384-at-eagle.mrc.ac.za} To: Microscopy-at-anlemc.msd.anl.gov
We have a vast amount of experience with the processing of myocardial / muscular tissue. In our experience it is the phosphates found in muscular tissue which play a role in the formation of these dense deposits.
The essential factors in the formation of electron dense deposits appear to be phosphate buffer, ethanol and uranyl acetate. Glutaraldehyde may contribute in a way, while osmium does not appear to play a role. (See our article in STAIN TECHNOLOGY 65: 243-250 (1990). -------
Message-Id: {MAILQUEUE-101.931213084915.416-at-parmly1.parmly.luc.edu} To: Microscopy-at-anlemc.msd.anl.gov
Greetings, Has anyone had exerience with a diamond knife designed to cut semi-thin sections? That's correct, semi-thin sections, 0.2 - 5 um range. I have just seen an ad for a product from Diatome called the "Histo Knife" which claims to do just that. The main question would seem to be how many sections such a knife can cut before needing to be resharpened. Advice or comments welcome. I will post a summary of any private replies. Thanks in advance, Tobias Baskin
******************************** *************** Tobias I. Baskin /~~~\ Biol. Sci's * Univ. of Missouri c|o o\ Columbia, MO 65211 USA \ = / Tel:314-882-0173 """ FAX 314 - 882 - 0123 baskin-at-biosci.mbp.missouri.edu
I manage a biologically-oriented EM facility at a university. Service fees are supposed to cover all expenses although this has never happened. Actually, we probably could break even if we had 50-75% of full use. However, there are three fully-equipped labs on this campus which make this prospect unlikely. We charge university users $25/room hour for both TEM and SEM, neither of which are equipped with EDAX, etc. We also charge for film. However, sputter-coating, fixation chemicals, resins, grids are all supplied by me. Industrial users are charged $50/room hour. The scope charge is supposed to cover service contract, parts and all prep chemicals, etc. In practice, the SEM costs about half as much as the TEM to run (including sample prep expenses) so the SEM is subsidizing the TEM. This is a friendly and efficient way of running a lab since people aren't being constantly nickel-and-dimed and the function of the lab is kept focused on providing researchers with the most useful services and supplies. Neither is time wasted by billing users for minor items. Furthermore, clients with small projects need not shell out major bucks just to get basic supplies. We used to charge about $1/print to cover paper, chemicals, and rapidprocessor repairs. The darkroom has turned into a general-purpose college facility with the result that the college pays our darkroom expenses. Users buy their paper from me at cost.
Please keep this information absolutely confidential!
Rod Kuehn
On Fri, 10 Dec 1993, Margaret E. Hogan wrote:
} } I am trying to get a feel for the amount to charge for EM services. I recently } took over an EM facility, and was asked to revamp the pricing for service work. } I am interested in any pricing schedule, from private, commercial or university } settings. Any help in this project would be GREATLY appreciated. All info } will be held strictly confidential. } Thanks!! } } Peggy Hogan } The Jackson Laboratory } 600 Main Street } Bar Harbor, Maine 04609 } (207) 288-3371 #1450
I have have been using diamond knives for cutting epoxy embedded tissues for about 20 years. Nearly all of the sections I cut are in the range of 0.5 - 2.0 micrometers in thickness. I started using a diamond knife that was no longer satisfactory for EM work because of small scratches. The sections I have made for LM are superior in quality. I have made 10,000-15,000 (maybe more) sections on a single 3mm knife that originally cost about $300. This knife has saved me many hours of time that would have been devoted to making glass knives. I am now using another knife (4.0 mm) with a larger boat that make makes flawless sections. The older knife is still useful but it makes more scratches than it once did. But, a few scratches are insignificant for some applications.
I assume you have had lots of experience with glass knives and know how to use your microtome skillfully. Be careful to avoid striking the knife edge with the specimen when you set up the microtome. Don't take big bites. Start cutting with specimen block-faces that are no larger than about 0.25 mm on a side (truncated pyramidal shape) and gradually work up to 0.5 mm on a side as you cut. Larger sections can be cut (if essential) after you have gained experience. Clean the edge (front and back) with a small, wetted wedge-shaped piece of polystyrine foam held with forceps. Wipe only parallel to the edge. It is remarkable that even after cutting many semi-thin sections the knives I use for LM will still cut nice looking silver and gold sections although I don't usually use these. I have never used Diatome "Histo Knifes" but I am confident that they would be satisfactory.
I bet there are many diamond knives siting around in drawers in laboratories that need to be resharpened for EM work. Before buying a new "Histo Knife" I suggest that you try to obtain a used diamond knife from someone who has given up EM work or has set aside of knife because of the high cost of resharpening.
I am interested in buying an old diamond knife for use in cutting sections for light microscopy. Anyone who is willing to part with an old, but unused favourite please contact me soon.
Bob
Bob Ridge Biology ICU 10-2 Osawa 3-chome Mitaka-shi Tokyo 181 Japan
Reply_ RE} } Greyscale Printers Reply from: John Mansfield North Campus Electron Microbeam Analysis Laboratory University of Michigan 2455 Hayward Ann Arbor MI 48109-2143 Phone: (313)936-3352 jfmjfm-at-engin.umich.edu or John_Mansfield-at-mse.engin.umich.edu I just invested in a Laserwriter Pro 630 to replace our aging IIg that had been upgraded from an NTX. At $1740 you cant complain! It is a 600dpi printer using a Canon EX engine. It makes a nice job of greyscale images. I printed a couple of wedges that Mark Disko sent me and It seems to do aboout 60 grey levels that I can count. I plan to use it for student research notebook printing, much cheaper than Polaroid! Faculty like it as it gives good images at a fraction of the cost! We use it for digital images collected from the TEMs, ESEM and AFM on to Apple Macs. We have the DI AFM connected to Appletalk and the software thinks it is printing directly to LPT2. Works well. I have seen output from the Hewlett Packard 600 dpi printers too and they are also excellent, maybe even a little better. I was driven by price considerations here tho' and the Apple was little cheaper and directly replaced our old printer with no additional network connection problems. Just my few cents worth. John Mansfield.
Reply_ RE} AFM to Appletalk question Reply from: John Mansfield North Campus Electron Microbeam Analysis Laboratory University of Michigan 2455 Hayward Ann Arbor MI 48109-2143 Phone: (313)936-3352 jfmjfm-at-engin.umich.edu or John_Mansfield-at-mse.engin.umich.edu
--------------------------------------
Would you mind providing some information on how did you connect your DI AFM to Appletalk (software and hardware).
Thank you,
Raul Fainchtein fain-at-aplcomm.jhuapl.edu
I am sending this to the list as I think there may be others who are interested in this. Since the AFM is run by a PC and our lab is still mainly Mac based as far as computers are concerned, we have pinters and file servers set up for the Macs, but little support for the PC. Therefore to print and store files on a server we purchased an ethernet card for the PC. We used a 3Com card, a 3C503, a really cheap basic card (ISA). We then bought the Farallon PhonenetPC software. Farallon bought the AppletalkPC software rights from Apple a number of years ago and changed the name. Farallon make a number of networking products, including Timbuktu which allows remote control of Mac and Windows PCs from remote machines of either type (scary seeing a Mac with a full windows interface!! and vice versa!!). The Phonenet software runs on the ethernet card (runs on almost any card these days) and supports multiple protocols, Appletalk, IPX and TCP/IP. You can have these loaded simulataneously, but it eats into the PC memory and the SPM software runs in reduced memory mode when the networking software is loaded on our 8 meg machine. DI have told us we can upgrade to 12meg (max) but we need new ROMs for the SPM hardware. This may help and we are in the process of ordering the upgrade. We hope we could run the full blown version of the SPM software and the networking software too. We could then try running ftp and telnet from the PC to enable wide area file transfer (I plan to try NCSA Telent for the PC). When the Appletalk stack is loaded (we use a number of cute little batch files to reboot the machine with different software configurations loaded --- A digression here, we have a 21M floptical and some of it's software is incompatible with the network software and so beware. Also it is very slow, if youy are thinking of getting one, my advice is don't! Pop the extra cash and buy a 128M mag-opt instead, much better bang for the buck! ---).
So as I say, when the Appletalk stack is loaded you can run a small program called DA which is a little like the Apple Chooser and you can then connect to Appletalk file servers and Appletalk printers and use either postscript or Epson emulation. We use postscript although it is very slow. Part of the reason for this is that the postscript spool file created by the SPM software is huge. It is a bitmap dump of the image screen of the microscope. Instead of using postscript to draw the text,lines and boxes on the screen and then merely bitmapping the image area, the whole screen is bitmapped so a large proportion of the spool file is either black or white space. This is why the text on DI SPM images that are printed on a postscript printer has jagged edges, the text is part of teh bitmap and not postscript fonts. Although I have been chastised in the past for implying that DI do some things on the cheap, I do think that the way the postscript printing has been implemented was a shortcut that was never cleaned up. For presentation purposes it would be nice to have the text of the image be printed in true postscript fonts. This would seriously cut down on the spool file size. I have asked about this in the past and was given the impression that it was a low priority item. Note: THIS IS MY PERSONAL OPINION AND SHOULD NOT BE TAKEN AS THE OPINION OF MY EMPLOYER. Anyway, inspite of this minor criticism, the Appletalk software and ethernet card allows us to share resources effectively. We print to our LaserWriter Pro 630 in our own lab and also to a Kodak Colorease PS300 in another building, saving us the cost of purchasing a dedicated printer for the SPM. We are currently investigating the possibility of using NetWare on the SPM to send print jobs to a NetWare file/print server which then spools the output to a number of printers. This may improve the speed and throughput.
Hope this helps, feel free to ask further questions if any of this is unclear. John Mansfield.
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} Greetings, } Has anyone had exerience with a diamond knife designed to cut } semi-thin sections? That's correct, semi-thin sections, 0.2 - 5 um range. I } have just seen an ad for a product from Diatome called the "Histo Knife" } which claims to do just that. The main question would seem to be how many } sections such a knife can cut before needing to be resharpened. Advice or } comments welcome. I will post a summary of any private replies. } Thanks in advance, } Tobias Baskin } We have used the Diatome Histo knife in our lab for several years (where have you been?). They will cut about a gazillion sections then after they are resharpened they will cut a gazillion more.
R. A. Harris EM Facility Evolution and Ecology Univ. of Calif. Davis
I have already posed the following question to the NIH-IMAGE forum, but I would also like to know what members of this forum might suggest. I'm using an annular light ring, attached to my objective lens, to project a direct beam of white light onto my subject, which reflects some of that light back to my objective lens and video camera. This annular light ring isn't perfect in that it causes non-uniform illumination of my subject; in particular, the center of my subject is brighter than average and the edges are darker.
I can, of course, correct my acquired images for shading irregularities by applying any of various digital procedures, but this approach is also not per- fect. For example, to obtain a reference image for the digital shading cor- rection, I use a standard gray card that is well out of focus, which eliminates problems of the heterogeneous surface of the card itself. Unfortunately, the pattern and strength of illumination non-uniformities is dependent on distance from the light source, and the shading pattern on the out-of-focus reference is different enough from that of my in-focus image that the digital correction is not acceptable.
I suppose I can alter the mathematics of my digital shading correction, but that would be an emperical approach that is not satisfying in general. I'm now wondering about inserting a diffusing lens (a ring, ordoughnut, filter) between my light source and subject to convert the direct beam into a diffuse, flat wash of light onto my subject, regardless of its distance to the light source. Has anyone tried this, and if so, will it actually work to solve my problem? Are there any other suggestions?
Thanks in advance,
Paul Sheppard Laboratory of Tree-Ring Research University of Arizona, Tucson GRAD12-at-CCIT.ARIZONA.EDU
The last message I just saw posted on Microscopy Email forum about conducting an EMail campaign directed to the US Government in support of Basic Energy Sciences was not appropriate to this listserver.
Whether I agree or not with the message is irrelevant, the point is that it should NOT have been done without first clearing it with the SysOP, since the posting is not directly associated with Microscopy.
Clearing of messages being sent to a listserver when they DO NOT deal with the general subject matter is a common practice of Email etiquette, which is followed by most responsible users of systems such as this. Since this is an unmoderated listserver, each of us has the duty to police the system appropriately and I would like to remind you each of that obligation.
I will resend the list of rules to the person responsible ldm-at-apollo.numis.nwu.edu (L. D. Marks) for cluttering up the Microscopy Forum with the message.
We are in the process of evaluating LM/Digital Imaging systems for materials research and I would be grateful for any feedback on systems that are in use with regards to ease of use, capabilities, software support, etc. Thanks in advance...
Hi, To add further to the conversation about printers...Has anyone had experience using 1200 dpi laser printers to generate working (i.e. lab notebook-type quality) printouts for use in data acquisition of microscopic images (e.g. determining lengths of dendrites, counting objects). Of course, the images aren't good enough for publication, but the price of these printers is declining (I think I saw one in PC Magazine for about $2500) and the $/page is negligible. Thanks for any comments. Nancy Desmond Neurosurgery Univ. of Virginia ' Charlottesville, VA 22908 804.924.5607 (voice)
I'd like to elaborate on Ron Anderson's comments about how the LogE enlargers work. The newer versions (e.g. EM55) do not have condenser lenses, and the CRT is located directly above the negative, thus eliminating the need to keep any condenser lenses free of dust. The rastered spot on the CRT illuminates only that portion of the negative which is directly below the spot. Also, the photo detector does not measure light reflected from the printing paper, rather the beam splitter (fractionally-silvered mirror), which is below the negative and above the enlarging (objective) lens, reflects a portion of the light which has passed through the negative to a PMT. Thus the PMT and its circuitry measures the relative density of each portion of the negative and adjusts the brightness of the beam spot and its raster speed within a small range in an attempt to print all of the densities within the limited gamma of the printing paper. The electronics also allow the user to adjust or override the exposure decisions made by the enlarger.
Russell E. Cook Electron Microscopy Center for Materials Research Argonne National Laboratory Argonne, IL
} On a number of occasions, I have seen answers to posted notices but } I did not see the original posted notice. As an example, the current } discussion on autofill LN2 systems. The first message I saw was Ron } Anderson's response. I never saw the original message. It has happened on } other occasions. Any thoughts? My main concern is that I may be missing } topics of intrest or replies to messages I've posted.
The problem in many cases is on the recieving end not the posting end. The Mailserver frequently get messages that say something like:
"address not active, "machine is off-line" "address cannot be reached after N-days"
and several other variations. Some can be traced to the fact that some users turn their computers off and the various relay systems will only hold messages so long before they trash them. Others are due to network problems where a NameServer goes off-line etc..
This listserver trys several times to send a message through in case links go down, however, things donot always make it through. Sometimes the backlog of "Requeued Mail gets very large" and I have to clean out the backlog, otherwise the entire system comes to a halt.
You should remember that the mailing list is now OVER 500 subscriptions long and this takes a fair amount of negotiation. I avoid "erasing" names from the subscription list when there are problems as usually the networks address problems clear up in a day or so. If I see messages to the same host bouncing continuously for many days, then I remove that one from the list.
All the messages sent to the list are archived, but I have not gotten around to setting up a review/list etc.. At the moment I just don't have the time. SysOp for this List is only one of the HATS I wear here at ANL, and most of the development work has to be done on the cheap and off-hours at home after my kids get all their homework done and get to bed (Yawn)...
I've also been having abit of hardware problems lately with the main computer which server the ANL EMCenter and this listserver. I'm in the process of replacing it (at least the Purchase Order was filed a few days ago). It will likely take about a month or two (we are a Gov. Lab with all its paperwork) to get it in, installed, and debugged. I plan on trying to make the change over around late Feb, however, Murphy says it won't be that easy and/or simple.
In the interim, the list will have to bear with the glitches.
Sorry-- Nestor Z.
Right now maybe I can get back to my scope and get some work done! I've got some nice angle resolved, energy filtered, dispersion surfaces of graphite measured. Looks really great. Anyone else interested in that sort of thing???
This is in reference to the current topic of Liquid Nitrogen Auto-fill devices. We have never used an LN2 Auto-fill device on our system and from reading all the responses to this topic, I don't think that we will ever get one. I only need to fill our dewar twice a week. First thing Monday morning and on Friday afternoon before I leave for the weekend. We've never had any problems doing it this way. It seems like more trouble than it's worth. I just thought that I would put in my two cents.
We are now up to 4 Diatome Histo Knives. We serial section 4 mm long vestibular and auditory organs, decalcified temporal bones and cochlea, fetal stuff without decal at .5 to 5 microns, 25 micron methacrylate, etc. the oldest is 5-6 years old. Sharpening is largely a function of how often they get dropped, people overlap their dumonts when cutting dry, or cutting bone. They have paid for themselves many times over compared to the section quality and time cost of glass. Cutting cochlea or crunching through the otoconia of vestibular organs raises hell with glass but doesn't faze the histo knives. We would have used old thin sectioning diamonds, but didn't have any. We still use a lot of glass, especially for potentially destructive samples and for teaching. With the newer LKB knifemaker, we have been using thicker glass, which makes glass more attractive.
} } On Mon, 13 Dec 1993, Tobias Baskin wrote: } } } Greetings, } } Has anyone had exerience with a diamond knife designed to cut } } semi-thin sections? That's correct, semi-thin sections, 0.2 - 5 um range. I } } have just seen an ad for a product from Diatome called the "Histo Knife" } } which claims to do just that. The main question would seem to be how many } } sections such a knife can cut before needing to be resharpened. Advice or } } comments welcome. I will post a summary of any private replies. } } Thanks in advance, } } Tobias Baskin } Glen MacDonald Hearing Development Laboratories RL-30 University of Washington Seattle, WA 98195 (206)543-8360 glenmac-at-u.washington.edu
Message-Id: {MAILQUEUE-101.931216081620.288-at-parmly1.parmly.luc.edu} To: Microscopy-at-anlemc.msd.anl.gov
} Date: Wed, 15 Dec 1993 12:48:38 -0600 (CST) } From: "Nestor J. Zaluzec (708)-252-5075, -4964" {ZALUZEC-at-anlemc.msd.anl.gov} } To: Microscopy-at-anlemc.msd.anl.gov } Cc: ZALUZEC-at-anlemc.msd.anl.gov } Subject: Missing Messages:
} To All Subscribers on Microscopy Forum: } } On Dec. 15. L.Sutter wrote: } } } On a number of occasions, I have seen answers to posted notices but } } I did not see the original posted notice. As an example, the current } } discussion on autofill LN2 systems. The first message I saw was Ron } } Anderson's response. I never saw the original message. It has happened on } } other occasions. Any thoughts? My main concern is that I may be missing } } topics of intrest or replies to messages I've posted. } } The problem in many cases is on the recieving end not the posting end. } The Mailserver frequently get messages that say something like: } } "address not active, } "machine is off-line" } "address cannot be reached after N-days" } } and several other variations. Some can be traced to the fact that } some users turn their computers off and the various relay systems } will only hold messages so long before they trash them. Others are } due to network problems where a NameServer goes off-line etc.. } } This listserver trys several times to send a message through in case links } go down, however, things donot always make it through. Sometimes the } backlog of "Requeued Mail gets very large" and I have to clean out } the backlog, otherwise the entire system comes to a halt. } } You should remember that the mailing list is now OVER 500 subscriptions long } and this takes a fair amount of negotiation. I avoid "erasing" names from } the subscription list when there are problems as usually the networks } address problems clear up in a day or so. If I see messages to the } same host bouncing continuously for many days, then I remove that one } from the list. } } All the messages sent to the list are archived, but I have not gotten } around to setting up a review/list etc.. At the moment I just don't have } the time. SysOp for this List is only one of the HATS I wear here } at ANL, and most of the development work has to be done on the cheap and } off-hours at home after my kids get all their homework done and get } to bed (Yawn)... } } I've also been having abit of hardware problems lately with the } main computer which server the ANL EMCenter and this listserver. } I'm in the process of replacing it (at least the Purchase Order was } filed a few days ago). It will likely take about a month or two } (we are a Gov. Lab with all its paperwork) to get it in, installed, } and debugged. I plan on trying to make the change over around } late Feb, however, Murphy says it won't be that easy and/or simple. } } In the interim, the list will have to bear with the glitches. } } Sorry-- Nestor Z. } } Right now maybe I can get back to my scope and get some work } done! I've got some nice angle resolved, energy filtered, dispersion } surfaces of graphite measured. Looks really great. Anyone else } interested in that sort of thing???
I've been having lots of trouble lately with undeliverable mail (Nestor: same as last time--then, one of the elves in Computer Headquarters had done a partial recompile of the system. This time...?) So this is a test to see if THIS works... But also...I'm not a materials person, but your nice graphite makes me recall an article from 2 or so years ago where some folks did STM on graphite & demonstrated things like DNA, proteins, etc.. Except, they were using just graphite. (I also haven't seen their work mentioned by anyone else.) The question is, do such structures show up in your preps in the TEM? Phil Oshel
I am new to this forum, and new to microscopy in general. We have recently set up equipment in our lab to study the microstructure of continous fiber reinforced composite materials. In specific we are trying to characterize the "fiber waviness" in composite laminates and cylinders. We are trying to view the fibers in plane (longwise) in hopes of characterizing the observed with some battery of FFT analysis, but this comes with many challenging hurdles, such as intermittent fiber breakages and also 3-D fiber curvature effects which sometimes take the fibers out of view. We are also trying to perform cross-section measurements, sort of a statistical sampling really.
We are grappling with a couple of issues. Optimum specimen preparation for light microscopy, we doing sort of modified metallographic grinding and polishing - but we still have trouble with too much fiber fracturing possibly linked back to the very first cut we did on the material. Anyone who has removed coupons of compostie material from laminates for microscopic evaluation, can you suggest a cost effective technique for this. Our laminates start at 1/8" thich and go up to almost 1/2" thick - presently we use diamond sectioning blades from Metallurgical Supply in Houston mounted on a milling machine at 1400 rpm with thwe table on slowest pass to cut the plate into strips and then we bring the strips to our Buehler Isomet 2000 to make small chips to pot, grind, and polish. This requires a skilled operator in the machine shop, it is very time intensive and still we have fiber fracturing. Is there anyone out there using as tabletop unit witha feed mechainsm like a table saw? Any suggestions?
We are also getting more surface relief thsn we can cope with in the final specimen. Our material is a polysulfone matrix with 7 micron diameter graphite fibers, the matrix is softer than the fibers so we get pitting very easily. We have been able to minimize this by diddling with the polishing sequence. If anyone has tackled this problem and has any insight we would appreciate it.
Thirdly we are using mostly brightfield illumination, but we don't get very good contrast. We are using a Leica MeF3-A metallograph and the contrast just isn't sharp enough or "broad" enough for our subsequent image analysis needs - i.e. can't be thresholded easily. We have tried darkfield which appears to give us sharper contrast but over an even "tighter" range. Has anyone got any experience with this one?
bruce-at-macgate.di.com writes: (regarding dye sublimation printers) } These printers are a sharable resource. Connect your whole department to } it (and get them to help pay for it)
I agree, but do be careful when hooking a dye sublimation printer to a network. Too many times I have had to shut ours down because someone down the hall or outside the building has neglected to change the Chooser setting before submitting a lengthy text file.
Yet another UK microscopy meeting to add to the comprehensive list sent by NJZ on November 9th.
8th September 1994 High Resolution Transmission Electron Microcopy Institute of Physics Headquarters, Belgrave Square, London, UK
This one-day meeting is being organised by the IOP EMAG group in conjunction with the Institute of Materials Metal Science Committee to discuss recent developments in the techniques and applications of HREM. The meeting will consist of overviews by invited speakers including Prof. C.J. Humphries and Dr. J. Hutchison, followed by contributed talks. Prospective speakers are invited to submit abstracts of {250 words before 31st May 1994 to M. Aindow, at the address below, indicating a preference for a 12 or 25 minute talk. Further details will be available from the IOP meetings department shortly.
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.
The mailer appeared to bounce my last attempt to post this - my apologies if you receive two copies of the same message;
Yet another UK microscopy meeting to add to the comprehensive list sent by NJZ on November 9th.
8th September 1994 High Resolution Transmission Electron Microscopy Institute of Physics Headquarters, Belgrave Square, London, UK
This one-day meeting is being organised by the IOP EMAG group in conjunction with the Institute of Materials Metal Science Committee to discuss recent developments in the techniques and applications of HREM. The meeting will consist of overviews by invited speakers including Prof. C.J. Humphreys and Dr. J. Hutchison, followed by contributed talks. Prospective speakers are invited to submit abstracts of {250 words before 31st May 1994 to M. Aindow, at the address below, indicating a preference for a 12 or 25 minute talk. Further details will be available from the IOP meetings department shortly.
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.
This did not go out via the official route as the news.groups official posting has closed down for the holiday. I think we may have enough votes but just in case, if you know someone, with an interest in the newsgroup, who has not voted then please encourage them to do so. Thanks and compliments of the season to you all. John Mansfield. I have forwarded this to the confocal mailing list, the microscopy mailing list and the NIH-Image mailing list.
-------------------------------------- Second CALL FOR VOTES (of 2)
Unmoderated group sci.techniques.microscopy
Newsgroups line:
sci.techniques.microscopy The field of microscopy.
Votes must be received by 23:59:59 UTC, 2 January 1994.
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This vote is being conducted by a neutral third party. For voting questions only, contact pschleck-at-unomaha.edu. For questions about the proposed group, contact John F. Mansfield {John.F.Mansfield-at-umich.edu} .
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.
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Many years ago, at another institution, I used a naphthyl thio acetate esterase histochemical reaction to mark histocytes and noticed that pericytes seemed to react and be labelled. Could never find any refs for esterases in pericytes and we lacked the funds to exhaustively check it out. This might help.
Glen MacDonald Hearing Development Laboratories RL-30 University of Washington Seattle, WA 98195 (206)543-8360 glenmac-at-u.washington.edu
On Fri, 17 Dec 1993, Martin B Garment wrote:
} Does any one know of a light microscopy stain that will differentiate between } endothelial nuclei and pericyte nuclei in capillaries, or of an antibody marker } specific for pericytes? } Martin Garment, UW-Madison, Madison, WI } mgarment-at-macc.wisc.edu }
Subject: Time:12:58 PM OFFICE MEMO TEM- JEOL 1010 Date:12/21/93 I'm new to the list and not sure if this inquiry is appropriate for the types of topics discussed on this list. If this type of inquiry is too specific or inappropriate please let me know. We purchased a JEOL 1010 TEM about 1 year ago and have had continual problemns with photography on the scope. We have approx.7 experienced TEM users (} 6 years experience) and several less experienced users which are all experiencing the same problem. Simply, when looking at images on the screen they are in focus, but when we photograph them, it becomes a lottery as to whether the images on the negative are in focus. JEOL has been out several times and has given the scope a clean bill of health each time using standard calibration grids. At their last visit they used our specimans and experienced the same problem. They have concluded that the problem may be speciman related, however, we are skeptical since it is present for a variety of users using different resins, tissue types etc.. Further, when we take these grids to other facilities in the area ( a Philips and a JEOL 1200) we seem not to have this problem. Does anyone else have experience, good or bad with this scope, or have any suggestions as to what the problem might be? We have been told by the service representitives that we are the only 1010 installation in the Eastern United States (we are in New Haven, CT). Does anyone on the list in the have this scope in the Eastern service region for JEOL?
Mike Sect. of Neurobiology Yale Univ. School of Medicine New Haven, CT 06510 Mike_Schwartz-at-qm.yale.edu
Subject: Time:2:44 PM OFFICE MEMO JEM-1010 Reply Date:12/21/93 I have not had direct experience with the model TEM you mention, but have managed electron microscopy labs for a quarter of a century. Users traditionally blame their problems on the instruments, thereby putting the burden of proof back on the Lab Manager. In a case such as this, the best thing to do is to obtain a reliable specimen, and keep it on hand to check the instrument whenever such a problem arises. In that way, you know whether it is the instrument or the users' specimens. I would suggest the 'Image Checker' specimen (No. 10070) and/or the 'Resolution Standard' specimen (No. 10090) supplied by Fullam (although equally suitable specimens are certainly available from other companies that handle EM supplies). Also, be sure that the specimen holder is clean, and that the specimens are being mounted in it so that they are held firmly and make good thermal contact. If this approach doens't clarify the situation to your satisfaction, then you really do have a problem.
} I'm new to the list and not sure if this inquiry is appropriate for the types } of topics discussed on this list. If this type of inquiry is too specific or } inappropriate please let me know. We purchased a JEOL 1010 TEM about 1 year } ago and have had continual problemns with photography on the scope. We have } approx.7 experienced TEM users (} 6 years experience) and several less } experienced users which are all experiencing the same problem. Simply, when } looking at images on the screen they are in focus, but when we photograph them, } it becomes a lottery as to whether the images on the negative are in focus.
This kind of question is very appropriate, Mike.
I manage a lab that has, among other equipment, a JEOL 2000 EX II. I'm not sure what the similarities are with your scope, but I do know that there are things to watch out for with these computer-controlled machines. Many users of our lab don't seem to understand that the scope is similar to a 200 CX. A major difference is that a computer controls the lenses, deflectors, etc, and stores the settings digitally. The guts of the scope, however, is still a series of analog devices, and they do drift. The computer helps maintain better control, but it is not magic.
I think that it is very important that users understand how to do the daily alignment procedures. They are easy to do. We use a brief instruction sheet to help them. I tweak the alignment periodically, to keep it close at all accelerating voltage ranges.
I agree with what Wil said. I have come to trust the machine, until I can absolutely rule out the specimen as the problem. Multi-user facilities are difficult in this regard.
If there is a problem with the scope, I know that JEOL will work with you to fix it. Good luck, and please post again when you find the problem. The solution is just as important as the problem.
John chandler-at-lamar.ColoState.EDU Fort Collins, CO
Mike Schwartz's focusing problem Before trying to make suggestions,it would be helpful to have the answers to a few basic questions. 1. Do you use "holey grids" and examine carbon holes to correct for astigmatism? 2. Have you made a recent through focus series of micrographs of carbon holes? 3. Have you made a recent trough-focus series of micrographs of well stained sections of standard histological material? 4. What criteria do you use in evaluating the focus of your negatives? 5. Are you familiar with the practice or art of obtaining "critical underfocus"? RAC
On Tue, 21 Dec 1993 23:26:42 +0200, John Chandler wrote:
} If there is a problem with the scope, I know that JEOL will work with you } to fix it. Good luck, and please post again when you find the problem. } The solution is just as important as the problem. } } John chandler-at-lamar.ColoState.EDU Fort Collins, CO
I agree with John about Jeol to help solving the problem. However, I would add one checkpoint for you. Do you focus using the image wobbler? If so, do you usually have the Optimum Under Focus switch ON? We have in my management a JEM 100C, JEM 100SX and JEM 1200EX, which all have the OUF switch. I have had to switch OFF the OUF because it seemed to bring the images too far underfocus.
Bet regards for Merry Christmas and a happy new year to all microscopy- netters
Jouko Maki
- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 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
} I agree with John about Jeol to help solving the problem. However, I would } add one checkpoint for you. Do you focus using the image wobbler? If so, do } you usually have the Optimum Under Focus switch ON? We have in my } management a JEM 100C, JEM 100SX and JEM 1200EX, which all have the OUF } switch. I have had to switch OFF the OUF because it seemed to bring the } images too far underfocus.
Yes, I'd forgot about the OUF, because we leave ours turned off. I don't think, however, that this would be the cause of inconsistent focus problems.
As for performance of various OUF's on JEOL's, when I was in a different lab with two 100CX's, we always had the OUF on. We always got beautiful negatives with it up to about 20K mag. I learned to trust it. There was one setting; it was ON or OFF. The 2000EX has OFF and three ON settings, low, medium and high. the medium setting is supposed to be equivalent to the ON setting of the 100CX.
One of the reasons we don't leave ours on now is because our heaviest users are used to the wobbler on the Philips 400. I haven't used one of these for years, but they remind me that it is bang on, even at 100K mag. They still use the wobbler on the JEOL to get close, then fine focus by eye. I have to keep reminding them that the two are different and not to expect them to behave the same.
Cheers,
John chandler-at-lamar.ColoState.EDU Fort Collins, CO
At ANL we sometimes use a combination of crushed dry ice and then methanol in the cooling holder this get you to about -30C. Does your cooling holder have a heater? The Gatan models do, in that case just turn up the heater until you reach a stable temp. It's a bit of guess work but it sometimes is sufficient. The only problem with this method is if the heater oscillates turning on and off, then you get specimen drift.
I need help in determining the best way to prepare a sample for TEM analysis. The specimens are secretory vesicles isolated from yeast. They are in a 0.8M sorbital buffer, to prevent osmotic damage. I was given approximately 50uL of each prep. The goal is to see the ratio of vesicles to sheets of plasma membrane in the prep. The vesicles should be mostly lipid. I wondered about osmium vapor fixation on a formvar grid, or some associated "negative" stain technique. I don't have a lot of specimen to do a routine TEM prep. Any help/suggestions would be greatly appreciated. Thanks in advance. Randy Nessler rnessler-at-emiris.iaf.uiowa.edu
Mike You did not explain "out of focus." When your negative is examined with an 8x lupe, is the grain "stretched" from astigmatism or drift? Or is the grain just very large? Or are the images just lacking in contrast?
Drift problems could be aggravated by excessively long exposures or too powerful an electron beam. Long exposures could be forced by excessively thick sections for your kv. High intensities result from improper "emission" or "bias"/filament heating combinations.
High kv seriously reduce contrast for biologicals.
Contrast can be seriously degraded by underexposure (intensity x exposure time too short) or underdevelopment caused by use of exhausted or cool developer.
Some scopes have low image contrast. Biological samples on such a microscope look crisp only when seriously underfocused so the problem isn't apparent until the negatives are examined. A standard is an inherently more contrasty subject. If it looks good in the scope, it is correctly focused. A good way of checking this would be to examine a section containing a hole. Focus on the membranes, then check the optical fringes in the hole.
Alternatively, your microscopists (being accustomed to a different scope) may be shooting too close to true focus, which delivers a muddy, low-contrast image with biologicals.
Good luck.
Rod Kuehn
} Subject: Time:12:58 PM } OFFICE MEMO TEM- JEOL 1010 Date:12/21/93 } experienced users which are all experiencing the same problem. Simply, when } looking at images on the screen they are in focus, but when we photograph them, } it becomes a lottery as to whether the images on the negative are in focus. } JEOL has been out several times and has given the scope a clean bill of health } each time using standard calibration grids. At their last visit they used our } specimans and experienced the same problem. They have concluded that the } problem may be speciman related, however, we are skeptical since it is present } for a variety of users using different resins, tissue types etc.. Further, } when we take these grids to other facilities in the area ( a Philips and a JEOL } 1200) we seem not to have this problem. Does anyone else have experience, good } or bad with this scope, or have any suggestions as to what the problem might } be? We have been told by the service representitives that we are the only 1010 } installation in the Eastern United States (we are in New Haven, CT). Does } anyone on the list in the have this scope in the Eastern service region for } JEOL? } } Mike } Sect. of Neurobiology } Yale Univ. School of Medicine } New Haven, CT 06510 } Mike_Schwartz-at-qm.yale.edu } }
Subject: Time:1:28 PM OFFICE MEMO JEM-1010 reply2 Date:12/22/93 In describing your problem, you simply stated that the images appeared to be "out of focus". This is a very specific problem that should only result from a variation in the current of one of the image-forming lenses, from variation in the high voltage, or from such effect as that of the OUF device, or perhaps from variation in the current in some device such as a set of deflecting or stigmator coils. All of these are electronic problems, and it should be possible for service engineers to check them out (i.e. to see if all likely circuits performing to specifications. UNSHARP images can be caused by a variety of other factors, however. As already suggested by others, specimen drift is a very common cause. For a discussion of sources and methods of diagnosing such instabilities you might want to read over Ch. 9 (Instabilities) in the book by J.C.H. Spence "Experimental High-Resolution Electron Microscopy", and Ch. 5 (Checking the performance of an EM) in the book "Principles & Practice of EM Operation" by A. W. Agar, et. al. which was published in the series "Practical Methods in Electron Microscopy" A. M. Glauert, Ed.
Grain-Boundary Characterization By Conventional TEM: A Survey Of Current Techniques
STUART MCKERNAN AND C. BARRY CARTER
Department of Chemical Engineering and Materials Science, University of Minnesota, Minneapolis, MN
ABSTRACT
The study of grain boundaries by transmission electron microscopy is now a mature field. Much of the research effort has focused on the analysis of a small set of special grain boundaries by high-resolution electron microscopy. More general boundaries necessarily have to be studied by other techniques. This paper will review the techniques currently available for the characterization of these grain boundaries and consider lines of future research.
National Center for Electron Microscopy, Lawrence Berkeley Laboratory, Berkeley, CA
ABSTRACT
This paper reviews recent progress in high resolution electron microscopy (HREM) of internal interfaces in solids. A brief summary of generic interface features of interest is followed by specific examples of HREM investigations of grain boundaries and heterophase interfaces. Features of interest include interface roughness, rigid body displacements, faceting, steps or ledges, elastic displacement fields, atomic bonding, composition gradients and localized atomic relaxation into structural units. Two types of analysis, direct interpretation and comparison with image simulations, are applicable to different types of interface characteristics. Critical issues of current importance and recent developments in technique are outlined for asymmetrical and symmetrical grain boundaries and for metal-semiconductor and metal-ceramic interfaces.
Synchrotron X-Ray Topographic Studies of Grain Boundaries
FUPING LIU, IAN BAKER
Thayer School of Engineering, Dartmouth College, Hanover, NH
ABSTRACT
Synchrotron white beam X-ray topography (SWBXT) is a powerful tool for studying the evolution of structural changes in bulk crystalline materials. This paper outlines the use of SWBXT in the transmission Laue geometry for studying grain boundaries (GBs). The advantages and disadvantages associated with SWBXT are described with the emphasis on in situ observations of the response of GBs to applied stress, thermal treatment and dislocation impingement.
Compositional Analysis of Interfaces Using X-ray Spectroscopy
ERNEST L. HALL
GE Corporate Research and Development, Schenectady, NY
ABSTRACT
In this paper, we examine the use of x-ray spectroscopy in the AEM to study compositional changes at interfaces in materials. The proper sample and microscope conditions for optimum data collection are discussed. The determination of the spatial resolution of the analysis, and the effect of spatial resolution on the analytical results, are described. Methods for deconvoluting the electron distribution in the sample from the solute distribution are reviewed. The effect of the minimum mass fraction detectable on this type of analysis is shown. Finally, examples of the use of x-ray spectroscopy to measure equilibrium and non-equilibrium composition variations at grain boundaries and interphase interfaces are shown.
EELS at Buried Interfaces: Pushing Towards Atomic Resolution
P.E. BATSON,1 N.D. BROWNING,2 and D.A. MULLER3
1IBM Thomas J. Watson Research Center, Yorktown Heights, NY; 2Oak Ridge National Laboratory, Oak Ridge, TN; 3Department of Applied and Engineering Physics, Cornell University, Ithaca, NY
ABSTRACT
Recently available increases in the sensitivity of electron spectrometry has allowed usable EELS signals to be obtained with the 2 sized probe needed to produce annular dark field (ADF) channeling contrast at 100 KeV. Three applications of this performance are discussed: 1) bonding and electronic structure obtained at a Si/SiO2 interface, 2) elemental Co composition at a CoSi2/Si interface, and 3) imaging using the and carbon transitions at a diamond/Si interface.
Atom-Probe Field-Ion Microscope Studies of the Chemistry of Internal Interfaces on an Atomic Scale
DAVID N. SEIDMAN, BRUCE W. KRAKAUER AND DAVID K. CHAN
Materials Science and Engineering Department and the Materials Research Center, Northwestern University, Evanston, IL
ABSTRACT
We explain the basic physical principles of both the field-ion and atom-probe microscopes with an emphasis on the atomic-scale imaging of atoms, and the concurrent measurement of the chemical identities of individual pre-selected atoms--on the surface of a field-ion microscope specimen--by time-of-flight mass spectroscopy. The advantages and disadvantages of atom-probe microscopy for the study of interfacial chemistry are enumerated. It is shown how an individual internal interface may be prelocated employing transmission electron microscopy, and then the same interface studied via atom-probe microscopy. The atomic-scale spatial-resolution capability of the atom-probe technique for studying interfacial chemistry is illustrated with two specific examples. The first one involves the direct and absolute measurement of the Gibbsian interfacial excess of solute at a grain boundary--a homophase interface in the Fe(Si) system; the depth resolution for the Si solute atom profile associated with the grain boundary is } 0.07 nm. The second application is the study of a metal/ceramic heterophase interface--the cadmium-oxide/silver {222} interface. It is demonstrated, via atom-probe microscopy, that the terminating {222} plane of CdO is the anion plane and not the cation plane; this result corresponds to a depth resolution of 0.136 nm and 0.118 nm in cadmium-oxide and silver, respectively, along a {111} direction. These examples illustrate the unique capabilities of an atom-probe to make standardless and quantitative measurements of interfacial chemistry on an atomic scale.
Three-Dimensional Atom Probe Studies of Solid-Solid Interfaces
THOMAS F. KELLY1,2,3, PATRICK P. CAMUS2, DAVID J. LARSON2,3, AND LOUIS M. HOLZMAN2,3
1Department of Materials Science and Engineering; 2Applied Superconductivity Center; 3Materials Science Program; University of Wisconsin, Madison, WI
ABSTRACT
The origins and underlying concepts behind the three-dimensional atom probe (3DAP) are described. Application of the 3DAP to the study of solid-solid interfaces is discussed in terms of the fundamental limitations of this technique compared with those of other characterization techniques, especially the conventional atom probe. Several examples of actual images from existing 3DAPs are shown with emphasis on their relevance to the study of solid-solid interfaces. Future developments in this form of microscopy which might have beneficial impact on the study of solid-solid interfaces are discussed.
The Tangent Formula in Electron Crystallography--Phase Determination of Copper Perchlorophthalocyanine
DOUGLAS L. DORSET1, MARY P. MCCOURT1, JOHN R. FRYER2, WILLIAM F. TIVOL3, JAMES N. TURNER3
1Electron Diffraction Department, Medical Foundation of Buffalo, Inc. Buffalo, NY; 2Electron Microscope Centre, Chemistry Building, University of Glasgow, Glasgow G12 8QQ, Scotland; 3Wadsworth Center for Laboratories and Research, New York State Department of Health, Albany, NY
ABSTRACT
The tangent formula as an appropriate method for phase determination in electron crystallography was evaluated using 198 experimental intensity data from ca 100 thick, epitaxially-oriented microcrystals of copper perchlorophthalocyanine. If the basis set used for the analysis (here with QTAN) is too small, the phase determination is unsuccessful. This agrees with independent assessments in another laboratory with MULTAN or RANTAN. However, a basis phase set of 27 reflections obtained from evaluation of three- and four-phase invariant sums is sufficient to phase 137 reflections, yielding Eh maps at lowest NQEST values that are readily interpreted in terms of atomic positions. This is an even smaller basis set than the e.g. 47 reflections at 2 resolution obtained from the Fourier transform of electron microscope images, which were shown earlier to be adequate for a successful phase extension.
The History of The Development of The First High-Resolution Electron Microscope
Remembering the Knoll Research Team at The Technical University Berlin, 1927-1934.
MARTIN M. FREUNDLICH
ABSTRACT
The first high-resolution electron microscope was developed by Ernst Ruska working first in cooperation with Max Knoll and later with Bodo V. Borries at the High-Tension Laboratory of the Technical University of Berlin. Though the electron microscope (EM) became one of the greatest and most far reaching achievements of the twentieth century, Ruska had to wait 53 years before being honored with the Nobel Prize. It took this long to sort out the merits of Reinhold Rudenberg's claim to be the sole inventor of the EM. He applied for a patent just 5 days before Knoll reported on the first low-resolution EM.
IBM Research Division, Zurich Research Laboratory, Sumerstr. 4, CH-8803 Rschlikon
ABSTRACT
An introduction to the principles, applications and perspectives of magnetic force microscopy (MFM) is given. Selected examples from magnetic recording as well as from fundamental research in magnetism are presented to demonstrate the type of information presently obtainable by MFM. Factors determining resolution are briefly discussed and some future perspectives for this method are described.
} I am currently working with marine orgasnisms and would appreciate any } suggestions for decalcifing them for em studies. } } Happy Holidays to all! } } Phil Rutledge } Email: prutle1-at-gl.umbc.edu
For EM of auditory receptors we used a decalcifying procedure first published by Baird, Winborn and Bockman (Anat. Rec. 159:281-290, 1967). This was to get rid of otoconia and any remaining otic capsule.
Briefly, we fixed in 4.0% aldehyde in 0.2 M s-collidine buffer with 2.0% sucrose. After buffer washes, specimens were decalcified using 0.1 M tetrasodium ethylenediamine tetraacetic acid (Na4EDTA), pH 7.4 (adjusted using versene acid), with 4.0% glutaraldehyde. Decalcifier was changed every other day, until decalcification was complete, usually 4-6 days. This was followed by post-fixation in OsO4 and normal processing for TEM or SEM. I don't think choice of buffer will affect decalcification.
A friend used a similar, but much higher concentration of EDTA, for dental specimens. EM preservation was still excellent with the higher concentration.
Good luck and Happy Holidays!
John chandler-at-lamar.ColoState.EDU Fort Collins, CO
I've used Warshawsky's EDTA for mammalian cochlea, mammalian and avian temporal bones with good results for EM and LM EDTA-2Na 0.111M 41.3 gm NaOH 0.11 M 44 gm Take to 1 L pH should be 7.4 decal tissues at 4 deg. C and change the solution every couple of days. 10% EDTA works ok, takes more NaOH to get the pH back up, Mori (J. Histochem. Cytochem. has a glycerol/EDTA solution forimmunocytochemisty at the EM level.. All of these work, the Mori is more trouble, I mostly stick with Warshawsky's.
Glen MacDonald Hearing Development Laboratories RL-30 University of Washington Seattle, WA 98195 (206)543-8360 glenmac-at-u.washington.edu
On Wed, 22 Dec 1993, rutledge phil wrote:
} I am currently working with marine orgasnisms and would appreciate any } suggestions for decalcifing them for em studies. } } Happy Holidays to all! } } Phil Rutledge } Email: prutle1-at-gl.umbc.edu } }
On Wed, 22 Dec 1993 17:34:33 +0200, John Chandler wrote:
} Yes, I'd forgot about the OUF, because we leave ours turned off. } } One of the reasons we don't leave ours on now is because our heaviest users } are used to the wobbler on the Philips 400. I haven't used one of these } for years, but they remind me that it is bang on, even at 100K mag. They } still use the wobbler on the JEOL to get close, then fine focus by eye. I } have to keep reminding them that the two are different and not to expect } them to behave the same. } } Cheers, } } John chandler-at-lamar.ColoState.EDU Fort Collins, CO
Hi again,
In case your problem occurs with a standard specimen, e.g. holey carbon, I strongly suggest you should build up heavy pressure against JEOL service people to find out the reason. It can be even a faulty DAC in the OL control circuit. Before going further you have to explain the exact way you make the exposures - that means the whole procedure. There may be some typical pattern how every user operates and that may help when analysing the reason for the problem. Make a point-to-point list of the procedures and try to find if there is a step which all operators do and which could cause a misfocussing if there exists an electronic fault. I have faced some DAC-troubles in our 1200EX, which is less integrated than 1010. Another source of component trouble is the optocouplers which are used to change the control voltage. I had to change each of them to a more reliable type and that was a lot of work. I hope you can solve the problem with the help of JEOL-people. It only takes some time to get it done.
Best regards,
Jouko
- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 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
} Date: Wed, 22 Dec 1993 17:06:13 -0500 (EST) } From: rutledge phil {prutle1-at-gl.umbc.edu} } Subject: decalcification } To: microscopy-at-anlemc.msd.anl.gov
} I am currently working with marine orgasnisms and would appreciate any } suggestions for decalcifing them for em studies. } } Happy Holidays to all! } } Phil Rutledge } Email: prutle1-at-gl.umbc.edu } Which marine orgs? Crustaceans? (if so, try the crust- l-at-sivm.si.edu mailserver at the Smithsonian), molluscs? forams? etc. What have you tried? such as EDTA? Phil Oshel po-at-parmly1.parmly.luc.edu
Subject: Time:8:47 AM OFFICE MEMO JEOL 1010 cont. Date:12/23/93 My thanks to the many individuals which have offered suggestions concerning our focusing problem. We are now systematically performing a variety of the solutions/tests that many of you have recommended. Unfortunately, the holidays have intervened and we will not complete this analysis till after the the 1st of the year. I will let you know the results when we are done. Just a little clarification on the nature of the focus problems and our previuos attempts to remedy them. The negatives are not dramatically out of focus look as if there is some drift in the specimans as several of you have suggested. Our initial attempts to address this problem included using 60Kv versus 80Kv as the accelerating voltage, using the wobbler to focus versus focusing "by eye", turning the under focus on or off, using different spot sizes, testing different embedding media (Epon/Arraldite vs. Durcapan) and using slot versus mesh grids. The tissue we use is brain tissue and we have even tested whether the quality of fixation, i.e. lightly fixed for immunohistochemistry or well fixed, makes a difference. While several of these parameters aggravate the problem, as one might expect, none of them rectified the problem. However, it is safe to say that there are certain types of specimans that are less likely to photograph out of focus such as holey grids used for alignment. Although all of these tests suggest that there may be a problem with the specimans, we are disturbed that this problem does not show itself with other scopes. We have taken the "exact" same grids and photographed them on a Phillips and a JEOL 1200 within our building and not experienced these problems. We also had no problem with the JEOL 100S that we traded in for the more sophisticated and versatile 1010. It is also very difficult to assume user error since 4 of us have over 15 years of TEM experience. Although we have tried many solutions, as I said, many of you have provided additional approaches which we have yet to attempt and we are hopeful that one of these may provide a solution. Thanks again for all the advice, and I'll keep you posted on the outcome.
Mike Sect. Neurobiology Yale Univ. Sch. Med. Mike_Schwartz-at-qm.yale.edu
Looking into relative merits of wet etching and plasma etching glass from microelectronic devices. There appears to be a bias towards plasma etching as being cleaner and less destructive to metallization under glass. We have used wet etching (Buffered HF solution) without any noticeable effect on metallization. However, the wet etch is at times incomplete and some areas of the die may have residue remaining. Any comments would be appreciated.
Also I would appreciate any information on the commercial plasma etchers available and their relative merits. Cost considerations are primary.
Thanks.
Richard Sartore at RSARTORE-at-MONMOUTH-ETDL1.ARMY.MIL
US ARMY RESEARCH LABORATORY AMSRL-EP-RA FORT MONMOUTH, NJ 07703-5601
Subject: Time:2:08 PM OFFICE MEMO TEM JEM-1010 reply3 Date:12/23/93 I strongly disagree with Ron Anderson's comments that the on-going discussion of the problem with 'unfocused' images on the JEM-1010 is inappropriate to this forum. I have used JEOL TEMs for many years, and agree that they are splendid instruments. In fact, I have probably been one of JEOL's strongest supporters. HOWEVER, subtle problems can, and unfortunately do, occur on virtually every model instrument, and are likely to be more difficult to diagnose on the more sophisticated models. Furthermore, service engineers are only human, and consequently the attention they provide varies with their individual background, their personal problems at any given moment, and a variety of other factors. I have also encountered a couple of service engineers, admittedly EXCEPTIONS to the general rule, who were outright belligerent and only marginally competent. I have not seen anything in this series of comments that should be taken as detrimental to JEOL instruments in general. The fact that the service engineer obtains what he claims are good micrographs suggests that the problem is in the users specimens or their techniques. (If faulty technique in using the instrument is the problem, then a good service engineer should be able to be helpful in overcoming it.) The fact that good micrographs are obtained from the users' specimens on other instruments (some of which are JEOL) suggests a problem with this particular instrument. Although I do not know them personally, at least some of the users involved here apparently are microscopists with a great deal of experience, and they seem to be taking an OBJECTIVE approach to attempting to obtain help with what is to them a very vexing problem. I see nothing wrong with discussing methods of resolving a situation of this kind. In fact, this is the kind of thing a forum such as this can be most helpful in dealing with.
Subject: Time:3:09 PM OFFICE MEMO JEM-1010 reply2 Date:12/23/93 OFFICE MEMO JEM-1010 reply2 Date:12/22/93 In describing your problem, you simply stated that the images appeared to be "out of focus". This is a very specific problem that should only result from a variation in the current of one of the image-forming lenses, from variation in the high voltage, or from such effect as that of the OUF device, or perhaps from variation in the current in some device such as a set of deflecting or stigmator coils. All of these are electronic problems, and it should be possible for service engineers to check them out (i.e. to see if all likely circuits are performing to specifications. UNSHARP images can be caused by a variety of other factors, however. As already suggested , specimen drift is a very common cause. For a discussion of this and other sources and methods of diagnosing such instabilities you might want to read over Ch. 9 (Instabilities) in the book by J.C.H. Spence "Experimental High-Resolution Electron Microscopy", and Ch. 5 (Checking the performance of an EM) in the book "Principles & Practice of EM Operation" by A. W. Agar, et. al. which was published in the series "Practical Methods in Electron Microscopy" A. M. Glauert, Ed.
Subject: Time:11:02 AM OFFICE MEMO JEOL 1010 Ethics Date:12/24/93 I would like to respond to Ron Anderson's comments that the discussion of our focusing difficulties is unethical and detrimental to JEOL. Firstly, we are long time users of JEOL TEMs and purchased the 1010 on the basis of our satisfaction with our prior JEOL TEM and our feeling the JEOL service has always been exempelary. My queries to this forum were not intended to impune the well deserved reputation for high quality that JEOL has come to enjoy as a result of good service and product. Rather, we have been trying to determine, in anyway possible, how we might improve the performance of the 1010 using both JEOL's resources, as well as those of any other informed sources. This serves to beneifit ourselves, may be of use to service technicians from JEOL which may have had only limited experince with this relatively new machine and may prove useful to future and current users of the JEOL 1010. We would not have even put this issue before the group if we had not obtained excellent results with the same tissue, grids and magnifications on other JEOL and Philips TEMs in our facility. Perhaps the 1010 is more sensitive to speciman parameters, if so, it is important that we and others know this. On the other hand many members of this list have provide several excellent suggestions for things to look at with regard to speciman preparation and possible machine tuning which we and the JEOL people had not yet explored. We are hopeful that these may help us resolve our problems. Perhaps, many members of the list feel that it is unethical to discuss particular products. If so, this should be made a part of the rules governing the use of the list which are sent to new members such as myself. However, I feel that I have benefited from feedback on particular commercial products on other lists. This has often saved me from purchasing inappropriate or substandard equipment or products.
I think the JEOL discussion has run it's course, let's let it cool off!
Part of the percieved problems MAY be due to the fact that messages occassionally get lost/trashed and not all subscribers see EVERY message and hence all readers do not see the entire discussion (I'm working on trying to fix this it appears to be an intermittent problems which randomly affects the listserver, and not the same individuals every time!).
Let me add the following:
* It is appropriate to discuss instrument problems. * It is appropriate to discuss specific manufacturers. * It is not appropriate to abuse a specific manufacturer user, or agency. * I read every message and take most with a large grain of salt, remember the ultimate goal of this forum is to let Microscopists help each other, and no one is perfect not even me :-)
I think there was abit of over-reaction here, but that's okay once and awhile. I did not think the discussion had gone out of bound yet, both points of view had valid points about making sure things were covered and still to remind everyone that a rumor could be ruinous to a company, should it not be true.
So to end on a more cheery note:
--------------------------------------------------------------- The Microscopy Listserver wishes everyone a good holiday season ---------------------------------------------------------------
Nestor Z. ANL EM Center ===============================================================
Electron Beams, Ion Beams (with apologies to the authors of Jingle Bells)
Dashing down the column traveling at hundreds of K. Through the sample they go scattering along the way. Mag and Beam up high making screens glow bright Oh what fun it is to see - atoms- day or night.
Electron Beams, Ion Beams, Photons on the Way! Oh what fun it is to be, in the E-M-C to-day T-E-M's, S-E-M's, Op-ti-cal scopes too! Scru-ti-nizing matter is what we love to do. E-D-S., E-L-S., Auger spectra too! Analyzing data it's all in store for you.
Electron Beams, Ion Beams, Photons on the Way! Oh what fun it is to see - atoms- day or night. A-F-M's, S-T-M's, Confocal scopes abound. We're here to study matter - from the whole world all around. Microtomes, Diamond Wheels, Lectro-polishing too. all this prep equipment and answers to be found..
Electron Beams, Ion Beams, Photons on the Way! Oh what fun it is to see - atoms- day or night. C-C-D's, V-C-R's, Video screens glowing bright displaying lots of data far into the night. Macrographs, - Micrographs, - Computed pictures too! Far too many pixels - to know with, what to do!
Electron Beams, Ion Beams, Photons on the Way! Oh what fun it is to see - atoms- day or night. Happy Holiday's from Us - and a Prosperous New Year too!
-------------------------------------------------------------- All the crew of the ANL EM Center:
Russ, Ed, Stan, Bob, Roseann, Charlie & Nestor --------------------------------------------------------------
On Thu, 23 Dec 1993 17:37:59 +0200, Ronald M. Anderson wrote:
} This is really simple! If anyone can take sharp pictures using standard } specimens then the machine is working fine! JEOLs are great machines } but I don't think they are smart enough to realize whose specimens } have been loaded and produce sharp pictures for one set of specimens } and 'out of focus' pictures for another set. } If the *JEOL* people get sharp pictures on standard specimens and } out of focus pictures on your specimens, why the skepticism? } It's your specimens! Are your specimens well bonded to a small mesh grid? } } In any case, I question the ethics of holding this dialog in this open formum. } Jouko Maki and others are searching for what could be wrong with the } instrument when it is not at all clear that the instrument is involved } and that you have not received satisfactory attention from the JEOL people. } Many readers will forget, or not read, these fine points and will carry } away the notion that the JEOL instrument has focus problems, therby } impacting their sales. That isn't fair!
Excellent,
I was not reading the original text carefully enough. Anyway, it essential to find out the procedure how people are making the exposures. It is also possible to find the reason for unsharp pictures from that starting point. It is quite common even to experienced researchers to learn to "bad habbits" while taking very many pictures. It would be interesting to know what other microscopes they are using.
As a comment to the in-focus standard specimen pictures, I would myself try what has already been told - that is: take a biological sample which normally gives unsharp pictures, find a hole in the support film (which I believe you are using?). Take test pictures by focussing to the hole-edge and to the normal biological structures. Compare the results. If the first is O.K. and the second not, You have solved the case - it is the method of focussing.
I am the last person to say anything negative about JEOL microscopes - if anyone has understood me to misevaluate them, that has not been my purpose. I have been using JEOL-microscopes since 1968 and found them very reliable and easy to use. I have also only positive things to tell about JEOL- service. What I am afraid is, that many microscopists are not "speaking the same language" with the service engineers.
I sincerely hope I have not created any negative pressure against JEOL by analysing the possible sources for un-focussed images.
Hello all, I work in the electron microscopy lab at the University of Iowa. We are in the process of purchaseing a SPM package from one of the commercial vendors (DI, Park, Topometric, etc). I am interested in hearing any information or experiences anyone has had with any of these systems. Please email me or post to microscopy-at-anlemc.msd.anl.gov. Thank you, Randy Nessler rnessler-at-emiris.iaf.uiowa.edu
We have a problem with an extraction voltage power supply on an HB-501 and would like to have copies of schematic # 139C-984 and 139C-974 if you have these available. The main schematic for these drawings is 139C-213-1. If you have the drawings needed would you mind sending a FAX to me at 602-965-9004. We have 139C-213-1 but not the others. Thank you. John C. Wheatley Arizona State University 602-965-3831
My apologies to the person whom I am addressing with this request for having forgotten your name. You requested opinions on automatic print processors. Were you able to draw any conclusions from the responses that you received?
Have you made a decision on a purchase. Did any general consensus materialize concerning one machine over another?
In this same regard it seems that whem using an automatic processor one should also have a means of automatically controlling expossure as well
What are the experiences of those out there using automatic exposure systems? ****************************** Greg Erdos * * Director, ICBR EMCL * * University of Florida *
My apologies to the person whom I am addressing with this request for having forgotten your name. You requested opinions on automatic print processors. Were you able to draw any conclusions from the responses that you received?
Have you made a decision on a purchase. Did any general consensus materialize concerning one machine over another?
In this same regard it seems that whem using an automatic processor one should also have a means of automatically controlling expossure as well
What are the experiences of those out there using automatic exposure systems? ****************************** Greg Erdos * * Director, ICBR EMCL * * University of Florida *