Just some additional thoughts on material choice for apertures:
When you speak of a Pt aperture it is probably Pt/Ir. The iridium allows for easier machining. We do pure platinum and gold apertures but only when the application demands it.
Another important consideration is the number and sizes of a multi-hole strip or plate. We have put up to 30 holes, some as small as 1micron, in a single part. Since the manufacture of burr-free, concentric apertures requires that the holes be made individually, more holes and smaller sizes result in more material losses during production. Platinum costs can add up.
For ultra-small holes and multi-hole parts moly should be used whenever possible.
John Arnott Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com
} Please send resumes and direct inquiries to: rccieslinski-at-dow.com } } POSITION OPEN } } Polymer Electron Microscopist } The Dow Chemical Company } Midland, Michigan } } Dow’s Corporate R&D Analytical Science Laboratory has a professional level } opening for a polymer microscopist at it’s Midland Michigan location. The } primary responsibilities would be to design, conduct and interpret } electron microscopy experiments to provide key answers to questions } relating to the development, processing and the performance of various } materials. Good written and oral communication skills are essential. The } ability to work both independently and in a team environment is also } extremely important. } } Educational Requirements: } A Ph.D. or MS degree in a materials or chemical related discipline is } preferred, but candidates with in a biological science related discipline } and substantial materials EM work experience would be considered. } } Experience Requirements: } Experience operation of a transmission electron microscope; excellent } manual dexterity and visual acuity; demonstrated ability to acquire and } objectively interpret data, and hands-on experience in the preparation of } specimens for Electron Microscopy, including ultramicrotomy and } cryo-ultramicrotomy. } } Key responsibilities will include: } * Strong problem solving skills. } * Active participation in development and project work teams. } * Interpretation and documentation of work } * Compliance with safety and quality programs. } } Send resumes and direct inquires to: rccieslinski-at-dow.com } } Robert C. Cieslinski, Ph.D. } The Dow Chemical Company } Corporate R&D, Analytical Sciences } 1897E Bldg. } Midland, MI 48667 } Fax: (989) 638-6443 } Email: rccieslinski-at-dow.com } } The Dow Chemical Company is the second largest chemical company in the } world and a leader in science and technology solutions. Come visit Dow } careers on the web at http://www.dow.com/careers. }
Call for Papers 13th Scandinavian Conference on Image Analysis SCIA 2003 Göteborg, Sweden, June 29 - July 2, 2003 http://www.hh.se/scia2003
Please notice the new date
The ambition of SCIA is to bring together leading researchers in the field of image analysis and present an exciting and advanced scientific program reflecting the state-of-the-art of the discipline. SCIA 2003 is the thirteenth in a series of conferences arranged every two years. We look forward to your contributions to this long-standing tradition.
Hosted this year by the Swedish Society of Image Analysis (SSAB), the conference is sponsored by the International Association for Pattern Recognition (IAPR) and the Nordic chapters of IAPR.
Scientific Program
The scientific program will include contributed as well as invited papers. Accepted papers will be published in the conference proceedings (Springer LNCS). The first day of the conference will consist of special sessions, such as workshops and/or tutorials.
Contributions covering the topics below are solicited. However, all submissions addressing issues related to image analysis are strongly encouraged. The main technical areas are:
- Image feature extraction - Image understanding - Grouping and segmentation - Motion analysis - Texture analysis - Color analysis - Shape analysis - Computer Vision - Cognitive vision - Medical image processing - Measurement and quantification - Measurement and visualization - Image coding and compression - Multi-modal processing - Indexing and databases - Images and the 3-D geometry - Standards and best practices - Images and pattern recognition - Classification - Applications
Invited Speakers and Workshop/Tutorial Presenters (not yet complete) Ivar Austvoll, Stavanger University College (NO) Ewert Bengtsson, Uppsala University (SE) Lars Bååth, Halmstad University (SE) Herve Delingette, INRIA, Sophia-Antipolis (FR) Chris Glasbey, Biomathematics & Statistics Scotland (UK) Ed Hancock, University of York (UK) Ioannis Kakadiaris, University of Houston (US) Rasmus Larsen, Technical University of Denmark (DK) Jussi Parkkinen, University of Joensuu (FI) Milan Sonka, University of Iowa (US)
Venue Situated in the heart of Scandinavia, Göteborg is within easy reach by air, rail and sea. There are daily flights from all the main European airports to Göteborg's Landvetter airport. The city center is only 20 minutes away by coach. Founded in 1829, Chalmers University of Technology is named after the major benefactor, William Chalmers, one of the directors of the successful Swedish East India Company in Göteborg. Today, the campus area with its modern conference facilities is situated in the city center close to hotels, restaurants, theatres, and shopping centers.
Social Activities Besides the conference dinner, there will be plenty of opportunities for social activities including boat cruises in the beautiful Göteborg archipelago, Liseberg - the largest amusement park in Scandinavia, the Botanic Garden, and many others.
Instruction to Authors Max 6 pages according to Springer LNCS format http://www.springer.de/comp/lncs/authors.html Look under the heading "Proceedings and Other Multi-author Volumes" describing format files and providing help. Papers should be submitted to: scia2003.papers-at-hh.se
Important Dates Submission of papers: January 22, 2003 Notification of acceptance: March 22, 2003 Camera-ready papers: May 8, 2003 Conference dates: June 29 - July 2, 2003
Contact Information http://www.hh.se/scia2003 SCIA2003-at-gbg.congrex.se
Conference Co-Chairs Josef Bigun Tomas Gustavsson
Programme Committee Fritz Albregtsen (NO) Kalle Åström (SE) Ivar Austvoll (NO) Ewert Bengtsson (SE) Ketil Bo (NO) Magnus Borga (SE) Gunilla Borgefors (SE) Stefan Carlsson (SE) Henrik Christensen (SE) Per-Erik Danielsson (SE) Bjarne Ersböll (DK) Robert Forchheimer (SE) Eric Granum (DK) Anders Heyden (SE) Jukka Iivarinen (FI) Peter Johansen (DK) Fredrik Kahl (SE) Hans Knutsson (SE) Björn Kruse (SE) Rasmus Larsson (DK) Reiner Lenz (SE) Tony Lindeberg (SE) Claus Madsen (DK) Henning Nielsen (DK) Ingela Nyström (SE) Erkki Oja (FI) Sören Olsen (DK) Matti Pietikäinen (FI) Ann Sohlberg(NO) Örjan Smedby (SE) Antanas Verikas (SE) Qin Zhong-Ye (SE)
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (little-at-earthtech.org) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, October 1, 2002 at 21:27:21 ---------------------------------------------------------------------------
Email: little-at-earthtech.org Name: George Luce and Scott Little
Organization: Earthtech International, Inc.
Education: Graduate College
Location: Austin, Texas
Question: We have recently revived a Cambridge Stereoscan 150 SEM from about 1978. The machine is now operational, but we cannot get good resolution or focus above a few thousand X magnification.
We would like to communicate with someone familiar with this or a similar model, who could give us some guidance on improving the resolution. Details of our efforts can be supplied. We are technically competent people with backgrounds in physics, engineering, high voltage, electronics, vacuum, etc.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mgrace-at-fit.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, October 1, 2002 at 15:16:59 ---------------------------------------------------------------------------
Email: mgrace-at-fit.edu Name: Michael Grace
Organization: Florida Institute of Technology
Education: Graduate College
Location: Melbourne, FL, USA
Question: I am working toward aquisition of a new SEM that will see use mostly in biological sciences applications, but will also see some use from materials scientists, chemists, etc. I am considering one of the new ESEMs (specifically the FEI/Philips Quanta 200. Question: how useful is the low vacuum mode, esp. for biological applications? Also interested in feedback from those with other applications, but I mainly want to be sure that the money is spent wisely- that we are not getting all the bells and whistles simply because they exist.
Yes, but are you experienced in electron microscopes? This is definitely a field that not only encompasses those mentioned, but uses them, as well as others, all in synergistic manner.
When it comes to resolution, look to your friend, the condenser control (or spot size or beam current or whatever else it may be called). On that vintage of Cambridge, I expect it is called spot size. If everything else is OK, the condenser is the primary control that determines the resolution of the instrument.
This is because the primary aberration in an electron optic instrument is chromatic aberration. That is the result of the wide spread in energy ranges of the electrons emitted from the electron gun. That spread creates what is known as a spherical aberration that affects the beam in both the X and Y directions. As they travel through the various electromagnetic lenses of the SEM, electrons of different energies are affected differently primarily resulting in a different focal point in the Z axis. The condenser lens, and associated apertures, are used to reduce this energy spread.
Here's the deal. The higher the condenser current, the smaller the spot size and the smaller the beam current. It's hard to explain in text, but you actually have two condenser lenses. Their purpose is to provide a de-magnification (reduction in apparent size) of the source of the electrons. The source is the tungsten filament, or cathode, that emits electrons from a rather large area of around 100 microns (micrometers).
In the process, there are a couple of imaginary images formed, or crossover points, where apertures or electron absorbers with small central openings are located. Electrons of different energies will be affected differently by the electromagnetic lenses - electrons of lower energy will be displaced more than those of higher energies. The apertures will capture those electrons that have an energy lower or higher than that which the lenses focus through the apertures, thus reducing the beam current by removing those electrons with energies outside of the nominal and reducing the spot size by ensuring that those electrons remaining have a smaller spread of energies.
Having said that, increase the condenser current, reduce the spot size or reduce the beam current, whichever applies.
If you're new to electron microscopy, there is one thing that really must be stressed - cleanliness. The smallest, microscopic, piece of dust, lint, fiber or electrically insulating contamination can wreak havoc inside an electron optics column. They tend to build up a negative electrical charge under influence of the beam that can create repulsive fields that deflect the electron beam and create weird aberrations. Contamination from fingerprints and solvents is extremely minor compared to the effects of a single fiber in the electron path.
Lastly, what ultimately determines the resolution possible in an instrument is the care with which it was installed. That includes the siting of the instrument in regards to mechanical vibrations in the building as well as the sources of electromagnetic interference. But most important is the careful routing of the various wires and tubing within and outside the instrument. The electron optics column is mounted on a table that is isolated from vibrations from the frame that holds it. All connections that run to this table must be carefully routed in order to provide similar isolation. Of similar concern is the electrical grounding of the various components of an SEM. Do not assume that because various large metal components appear to have electrical continuity that they do. Manufacturers typically provide for individual grounds to many components for good reason.
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174
On Tuesday, October 01, 2002 10:51 PM, by way of MicroscopyListserver [SMTP:little-at-earthtech.org] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (little-at-earthtech.org) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, } October 1, 2002 at 21:27:21 } --------------------------------------------------------------------------- } } Email: little-at-earthtech.org } Name: George Luce and Scott Little } } Organization: Earthtech International, Inc. } } Education: Graduate College } } Location: Austin, Texas } } Question: We have recently revived a Cambridge Stereoscan 150 SEM } from about 1978. The machine is now operational, but we cannot get } good resolution or focus above a few thousand X magnification. } } We would like to communicate with someone familiar with this or a } similar model, who could give us some guidance on improving the } resolution. Details of our efforts can be supplied. We are } technically competent people with backgrounds in physics, } engineering, high voltage, electronics, vacuum, etc. } } Thank you. } } } George Luce (geolucetx-at-aol.com) } } Scott Little (little-at-earthtech.org) } } } Earthtech International, Inc. } Austin, Texas } } Phone (512) 342-2185 } } www.earthtech.org } } --------------------------------------------------------------------------- }
In this case the "bells and whistles" are (still, I believe) unique as they allow you to look at fully hydrated specimens and have water in the chamber.
I am a happy XL30 ESEM (tungsten) user. If you can afford it the FEGESEM has advantages over the tungsten filament ESEM related to kV, spot size, SE signal and beam damage for viewing biological samples.
Dave
On Wed, 2 Oct 2002 00:53:13 -0500 by way of MicroscopyListserver {mgrace-at-fit.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (mgrace-at-fit.edu) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, } October 1, 2002 at 15:16:59 } --------------------------------------------------------------------------- } } Email: mgrace-at-fit.edu } Name: Michael Grace } } Organization: Florida Institute of Technology } } Education: Graduate College } } Location: Melbourne, FL, USA } } Question: I am working toward aquisition of a new SEM that will see } use mostly in biological sciences applications, but will also see } some use from materials scientists, chemists, etc. I am considering } one of the new ESEMs (specifically the FEI/Philips Quanta 200. } Question: how useful is the low vacuum mode, esp. for biological } applications? Also interested in feedback from those with other } applications, but I mainly want to be sure that the money is spent } wisely- that we are not getting all the bells and whistles simply } because they exist. } } Thank you. } } --------------------------------------------------------------------------- }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Has the confocal listserver been closed or have I been purged? Haven't gotten anything for a while. Gregory W. Erdos, Ph.D. Assistant Director, Biotechnology Program Scientific Director, ICBR EM CORE University of Florida Ph. 352-392-1295 PO Box 118525 Fax 352-846-0251 Gainesville, FL 32611 http://www.biotech.ufl.edu/EM
I was hoping someone could recommend some manufacturers of confocal microscopes. We are just beginning to investigate the possibility of purchasing one for our labs and while the literature I've read is intriguing, I would like more specific information about the various scopes available and what they can really do for us. Also, I need some idea of the price. I would appreciate it if any manufacturers or people with personal accolades for a specific scope would please contact me offline. Thanks for your help!
Would someone please direct me to some resource, a good text, or better yet, a web site that can help me identify wood types by either cross-grain and/or with grain using a microscope? I have some crushed slivers that I'd like to identify. Any help would be appreciated. Thanks in advance.
I have a JEOL JSM-5800LV, which is a low vacuum model. I use the LV feature quite a bit.
The down side is: you must use backscatter detection in this mode, a slight loss in high-mag resolution, and cost.
The plus side is convenience - samples don't have to be coated...they can be imaged as-is. There's no spurious EDS peaks from conductive coatings, which makes spectrum interpretation easier. Sample pump-down is even quicker.
A lot of people are going with this new generation of SEMs. You still have the option of imaging the regular way with these instruments.
My application is mostly material science. Being concentrated in biology, I think you have a stronger argument for buying an LV SEM. It would see a lot of use in other disciplines once other people learn of the instrument's capabilities.
Stu Smalinskas Metallurgist SKF Plymouth, Michigan
__________________________________________________ Do you Yahoo!? New DSL Internet Access from SBC & Yahoo! http://sbc.yahoo.com
Dear Michael, We recently purchased a new SEM and got the variable-pressure option, since all the manufacturers offer this now and it doesn't cost much more. Ours is a materials engineering department, but the microscopes are used by all sorts of university and commercial users, including biologists. I was pleasantly surprised with how useful the VP is for materials applications. Samples that are not high vacuum compatible, such as wet or oily stuff, samples that are impossible to coat or that cannot be coated because they are needed in their current state for something else or just things you want to look at very quickly, all work fine in VP mode, either by back-scattered electron mode or a special secondary electron detector for variable pressure. Apparently, with a cool or cold stage, the applications are extended even more. That is next on my wish list. I would not consider buying a conventional SEM without variable pressure option, just for the versatility it offers. I have no experience with the FEI ESEM, which goes to a much higher pressure than the variable pressure range of my microscope. At 12:53 AM 10/02/2002 -0500, you wrote: } } Email: mgrace-at-fit.edu } Name: Michael Grace } } Organization: Florida Institute of Technology } } Education: Graduate College } } Location: Melbourne, FL, USA } } Question: I am working toward aquisition of a new SEM that will see } use mostly in biological sciences applications, but will also see } some use from materials scientists, chemists, etc. I am considering } one of the new ESEMs (specifically the FEI/Philips Quanta 200. } Question: how useful is the low vacuum mode, esp. for biological } applications? Also interested in feedback from those with other } applications, but I mainly want to be sure that the money is spent } wisely- that we are not getting all the bells and whistles simply } because they exist. } } Thank you. } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Excellent point. And just some additional recommendations.
SEM resolution is highly specimen dependant. If you do not have standard specimens for checking resolution, you can try (for magnifications up to 10k) fractured metal. Do not use non-conducting coated specimens.
Use shortest possible working distance.
If you cannot get rid completely of astigmatism with electronic controls you should clean and/or realign column.
By the way, what is "a few thousand X magnification"? 10k could be pretty good for 1978 vintage SEM.
Vladimir
} Yes, but are you experienced in electron microscopes? This } is definitely a } field that not only encompasses those mentioned, but uses } them, as well as } others, all in synergistic manner. } } When it comes to resolution, look to your friend, the } condenser control (or } spot size or beam current or whatever else it may be called). } On that } vintage of Cambridge, I expect it is called spot size. If } everything else } is OK, the condenser is the primary control that determines } the resolution } of the instrument. } } This is because the primary aberration in an electron optic } instrument is } chromatic aberration. That is the result of the wide spread } in energy } ranges of the electrons emitted from the electron gun. That } spread creates } what is known as a spherical aberration that affects the beam } in both the X } and Y directions. As they travel through the various electromagnetic } lenses of the SEM, electrons of different energies are } affected differently } primarily resulting in a different focal point in the Z axis. The } condenser lens, and associated apertures, are used to reduce } this energy } spread. } } Here's the deal. The higher the condenser current, the } smaller the spot } size and the smaller the beam current. It's hard to explain } in text, but } you actually have two condenser lenses. Their purpose is to } provide a } de-magnification (reduction in apparent size) of the source of the } electrons. The source is the tungsten filament, or cathode, } that emits } electrons from a rather large area of around 100 microns } (micrometers). } } In the process, there are a couple of imaginary images } formed, or crossover } points, where apertures or electron absorbers with small } central openings } are located. Electrons of different energies will be } affected differently } by the electromagnetic lenses - electrons of lower energy } will be displaced } more than those of higher energies. The apertures will capture those } electrons that have an energy lower or higher than that which } the lenses } focus through the apertures, thus reducing the beam current } by removing } those electrons with energies outside of the nominal and } reducing the spot } size by ensuring that those electrons remaining have a } smaller spread of } energies. } } Having said that, increase the condenser current, reduce the } spot size or } reduce the beam current, whichever applies. } } If you're new to electron microscopy, there is one thing that } really must } be stressed - cleanliness. The smallest, microscopic, piece } of dust, lint, } fiber or electrically insulating contamination can wreak } havoc inside an } electron optics column. They tend to build up a negative } electrical charge } under influence of the beam that can create repulsive fields } that deflect } the electron beam and create weird aberrations. Contamination from } fingerprints and solvents is extremely minor compared to the } effects of a } single fiber in the electron path. } } Lastly, what ultimately determines the resolution possible in } an instrument } is the care with which it was installed. That includes the } siting of the } instrument in regards to mechanical vibrations in the } building as well as } the sources of electromagnetic interference. But most } important is the } careful routing of the various wires and tubing within and } outside the } instrument. The electron optics column is mounted on a table that is } isolated from vibrations from the frame that holds it. All } connections } that run to this table must be carefully routed in order to } provide similar } isolation. Of similar concern is the electrical grounding of } the various } components of an SEM. Do not assume that because various large metal } components appear to have electrical continuity that they do. } Manufacturers typically provide for individual grounds to } many components } for good reason. } } } } } Allen R. Sampson } Advanced Research Systems } 317 North 4th. Street } St. Charles, Illinois 60174 } } phone (630) 513-7093 fax (630) 513-7092 http://www.sem.com } } } On Tuesday, October 01, 2002 10:51 PM, by way of MicroscopyListserver } [SMTP:little-at-earthtech.org] wrote: } } } -------------------------------------------------------------- } ---------- } } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -------------------------------------------------------------- } ---------. } } } } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } } submitted by (little-at-earthtech.org) from } } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, } } October 1, 2002 at 21:27:21 } } } -------------------------------------------------------------- } ------------- } } } } Email: little-at-earthtech.org } } Name: George Luce and Scott Little } } } } Organization: Earthtech International, Inc. } } } } Education: Graduate College } } } } Location: Austin, Texas } } } } Question: We have recently revived a Cambridge Stereoscan 150 SEM } } from about 1978. The machine is now operational, but we cannot get } } good resolution or focus above a few thousand X magnification. } } } } We would like to communicate with someone familiar with this or a } } similar model, who could give us some guidance on improving the } } resolution. Details of our efforts can be supplied. We are } } technically competent people with backgrounds in physics, } } engineering, high voltage, electronics, vacuum, etc. } } } } Thank you. } } } } } } George Luce (geolucetx-at-aol.com) } } } } Scott Little (little-at-earthtech.org) } } } } } } Earthtech International, Inc. } } Austin, Texas } } } } Phone (512) 342-2185 } } } } www.earthtech.org } } } } } -------------------------------------------------------------- } ------------- } } } } }
The low vacuum mode is extremely useful for biological samples. For low magnification pictures it is extremely useful and these days we hardly use our sputter coater and critical point dryer, no fixation (you avoid use of hazardous fixatives in that way). We are very satisfied with the results. We have a Hitachi 3200N variable pressure SEM.
Good luck,
Soumitra Ghoshroy
************************************************************* Soumitra Ghoshroy Ph.D. Director, Electron Microscopy Lab Graduate Faculty Member, Department of Biology Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office), 646-3283 (lab) Fax: 505-646-3282 e-mail:sghoshro-at-nmsu.edu URL:http://confocal.nmsu.edu/eml
On Wed, 2 Oct 2002, by way of MicroscopyListserver wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (mgrace-at-fit.edu) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, } October 1, 2002 at 15:16:59 } --------------------------------------------------------------------------- } } Email: mgrace-at-fit.edu } Name: Michael Grace } } Organization: Florida Institute of Technology } } Education: Graduate College } } Location: Melbourne, FL, USA } } Question: I am working toward aquisition of a new SEM that will see } use mostly in biological sciences applications, but will also see } some use from materials scientists, chemists, etc. I am considering } one of the new ESEMs (specifically the FEI/Philips Quanta 200. } Question: how useful is the low vacuum mode, esp. for biological } applications? Also interested in feedback from those with other } applications, but I mainly want to be sure that the money is spent } wisely- that we are not getting all the bells and whistles simply } because they exist. } } Thank you. } } --------------------------------------------------------------------------- } }
FEI's ESEMs are equipped with good gaseous secondary electron detectors and for "good" specimens their resolution in wet mode is pretty close to the resolution in conventional mode. They are widely used in materials science.
Nevertheless, in biological applications most specimens are not "good". The best results I have had with hard tissue, dentin and bone. In wet mode with wet dentin or bone I can use magnification up to 25k, but if these specimens are dry then in the same conditions (wet mode) magnification is limited to 10k. With Au-Pd coated specimens in high vacuum mode I can use magnifications up to 150k (I have a field emission ESEM). I have had good results in observation plants and (surprise!) bugs. Success with cell cultures was marginal. And I have to mention that application of EDS analysis in wet mode could be complicated and limited.
However, with all these limitations, I have found that ESEM is a useful tool in our research.
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} Email: mgrace-at-fit.edu } Name: Michael Grace } } Organization: Florida Institute of Technology } } Education: Graduate College } } Location: Melbourne, FL, USA } } Question: I am working toward aquisition of a new SEM that will see } use mostly in biological sciences applications, but will also see } some use from materials scientists, chemists, etc. I am considering } one of the new ESEMs (specifically the FEI/Philips Quanta 200. } Question: how useful is the low vacuum mode, esp. for biological } applications? Also interested in feedback from those with other } applications, but I mainly want to be sure that the money is spent } wisely- that we are not getting all the bells and whistles simply } because they exist. } } Thank you. } } -------------------------------------------------------------- } ------------- } }
Following is a description of surplus Kevex equipment. If you are interested in any of these items and are willing to pay shipping expenses please contact me.
Regards, Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
1. Kevex DeltaPlus Microanalysis System, Level 5 (without detector) Serial #5500240-0158 Installed: November 1992 Electronics cabinet houses the signal processing electronics, MCA, 12-slot dual wide DEC Q-bus card cage with LSI-11/73 minicomputer, dedicated microprocessor and associated logic circuitry and data storage memory. Kevex model 4461A pulse processor. Single Syquist drive available.
2. Kevex Delta Analyzer, complete system, however, light element detector has 'blown' window Serial #501678-1172 Detector: 30mm square, model 3600-0674 Serial #0790-5803 Pre-amplifier model 2003 Installed: October 1990
To be very precise, crushed slivers may not be adequate, since some distinctions are based on the variations in cell shape that occur through an annual growth ring. You might want to contact Arno Schniewind :
arnops-at-nature.berkeley.edu
of the Forest Products Laboratory of the University of California, or his colleagues:
http://www.ucfpl.ucop.edu/
Arno has been very helpful over the years and the lab provides wood identification services.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Listers, } } Would someone please direct me to some resource, a good text, or better } yet, a web site that can help me identify wood types by either cross-grain } and/or with grain using a microscope? I have some crushed slivers that I'd } like to identify. Any help would be appreciated. Thanks in advance. } } Chuck Butterick } Degussa Engineered Carbons } Borger, TX
I would like to thank all the people for their helpful suggestions and knowledge concerning the quantitative composition analysis in a TEM.
I did not give the details clearly when I posted the question. I'd just like to provide more information on the detector and the microscope I used. The microscope is a Jeol-2010 LaB6 (though I wish it was a FEG) operated at 200kV. The detector is a PGT Ge detector with a ultra thin window. The software is called imix. The atomic fraction I got for the EuS is calculated automatically from this software after putting in voltage, take-off angle, density, and sample thickness.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Michael, We recently purchased a new SEM and got the variable-pressure option, since all the manufacturers offer this now and it doesn't cost much more. Ours is a materials engineering department, but the microscopes are used by all sorts of university and commercial users, including biologists. I was pleasantly surprised with how useful the VP is for materials applications. Samples that are not high vacuum compatible, such as wet or oily stuff, samples that are impossible to coat or that cannot be coated because they are needed in their current state for something else or just things you want to look at very quickly, all work fine in VP mode, either by back-scattered electron mode or a special secondary electron detector for variable pressure. Apparently, with a cool or cold stage, the applications are extended even more. That is next on my wish list. I would not consider buying a conventional SEM without variable pressure option, just for the versatility it offers. I have no experience with the FEI ESEM, which goes to a much higher pressure than the variable pressure range of my microscope. At 12:53 AM 10/02/2002 -0500, you wrote: } } Email: mgrace-at-fit.edu } Name: Michael Grace } } Organization: Florida Institute of Technology } } Education: Graduate College } } Location: Melbourne, FL, USA } } Question: I am working toward aquisition of a new SEM that will see } use mostly in biological sciences applications, but will also see } some use from materials scientists, chemists, etc. I am considering } one of the new ESEMs (specifically the FEI/Philips Quanta 200. } Question: how useful is the low vacuum mode, esp. for biological } applications? Also interested in feedback from those with other } applications, but I mainly want to be sure that the money is spent } wisely- that we are not getting all the bells and whistles simply } because they exist. } } Thank you. } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Hoadley, R.B. (1990) Identifying Wood: Accurate Results with Simple Tools. Taunton Books. 1990 (try Amazon.com) Parham, R. and Gray, R. (1982) The Practical Identification of Wood Pulp Fibers. TAPPI Press, 212 pp. 1982.(try www.tappi.org) Strelis, I. and Kennedy, R.W. (1967) Identification of North American Commercial Pulpwoods and Pulp Fibers. University of Toronto Press. 1967 Core, H.A., Cote, W.A., Day, A. C. (1979) Wood Structure and Identification. 1979
If you only have slivers, it can be difficult, expecially with conifers. I can recommend some experienced analysts off-line.
David Rothbard Earthborn Solutions rothbardD-at-netscape.net
} } Would someone please direct me to some resource, a good text, or better } yet, a web site that can help me identify wood types by either cross-grain } and/or with grain using a microscope? I have some crushed slivers that I'd } like to identify. Any help would be appreciated. Thanks in advance. } } Chuck Butterick } Degussa Engineered Carbons } Borger, TX
Could someone recommend a book, journal article or reference on the topic of liquid crystals that encompasses both an introduction to the topic and techniques for their characterization by light microscopy. Thanks.
Cheers, Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
I have been asked to process adult mouse tendon, a new tissue for me. I would appreciate any protocols or suggestions for processing this tissue from fixation through TEM plastic embedding. Also, should I be concerned about damage to my diamond knife during thin sectioning? Thanks in advance. Marilyn Levy
} Would someone please direct me to some resource, a good text, or better } yet, a web site that can help me identify wood types by either cross-grain } and/or with grain using a microscope?
R. Bruce Hoadley is an expert on wood identification. If your sample size is relatively large, i.e., splinter sized, rather than completely crushed, then you can probably get pretty far just from Hoadley's book _Identifying Wood_ which is still in print (Taunton Press) - about US$30 from Amazon.com. If not, you might try contacting Dr. Hoadley directly to see what he suggests.
In the past you have requested information on discounted products. If you are not a smoker, and find this email offensive, then we sincerely apologise. We will be only too happy to take you off our database.
If you are a smoker, however, you are probably fed up with paying high prices for your cigarettes and tobacco. Take a look at what we can do for you at http://www.smokersassociation.co.uk/?S=32&ID=2
We can send you, legally, by registered air mail, direct to your door, 4 cartons of cigarettes or 40 pouches of rolling tobacco (all brands are available) from only 170 Euros - about 105 pounds - fully inclusive of postage and packing. Why pay more?
If you would rather not hear from us any more, this link will ensure that you are not bothered again. mailto:unsubscribe-at-smokersassociation.co.uk
Microscopist and Lab Manager for a Centralized Electron Microscope Laboratory Facility at Portland State University
Portland State University seeks an electron microscopist to operate and manage a newly established electron microscopy (EM) facility consisting of a FEI/Philips (Tecnai F-20) 200kV field emission high-resolution transmission electron microscope (TEM) equipped with an embedded digital scanning transmission electron microscopy (STEM) capability, and energy dispersive x-ray spectrometer (EDS), a JEOL 2000FX TEM, and a 611 FEI focused ion beam microscope.
PSU is an AA/EO institution and, in keeping with the president's diversity imitative, welcomes applications from diverse candidates and candidates who support diversity.
The responsibilities of the position include the management, operation, and maintenance of the microscopes, and training and assisting faculty, and student, and outside university users of the microscopes.
Candidates preferentially have at least 5-years experience in managing an electron microscopy multi-user facility in an academic or industrial research setting and an outstanding record of team and personal accomplishments. A publication record of EM-related research is expected. Degree in electron microscopy and relevant disciplines in using microscopes is required.Advanced university degree is required as well as a demonstrated experiences in either materials science, geological, and or biological materials characterizations are essential. The candidates must have working knowledge onexperience using the microscopes listed above and are be familiar such operational techniques as digital image acquisition, image processing (digital micrograph) , structure simulation, and EDS analysis.
The candidates should also be able to have experience performing minor microscope repairs and maintenance. repair minor problems of the microscopes while mMicroscopes will be maintained under ajor components of the microscopes will be covered by the service contracts.
The successful candidates are expected to have be self-motivationmotivated, patience, and the ability to work and work well with exiting technical staff, faculty, and students, and outside users.
Candidates will be selected based on their qualifications and accomplishments. Salary will be commensurate with qualifications and experience.
Review of applications will begin on October 1, 2002, and continue until the position is filled.
Interested candidates, please should send a letter of interest with a two-page management plan for a multi-user EM facility and a full resume and arrange to have five letters of reference, sent to the address below:
Search Committee for EM Manager Physics DepartmentDepartment of Physics Portland State University P.O. Box 751 Portland, OR 97207-0751
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Sir or Madam
In the past you have requested information on discounted products. If you are not a smoker, and find this email offensive, then we sincerely apologise. We will be only too happy to take you off our database.
If you are a smoker, however, you are probably fed up with paying high prices for your cigarettes and tobacco. Take a look at what we can do for you at http://www.smokersassociation.co.uk/?S=32&ID=2
We can send you, legally, by registered air mail, direct to your door, 4 cartons of cigarettes or 40 pouches of rolling tobacco (all brands are available) from only 170 Euros - about 105 pounds - fully inclusive of postage and packing. Why pay more?
If you would rather not hear from us any more, this link will ensure that you are not bothered again. mailto:unsubscribe-at-smokersassociation.co.uk
I have been coating samples via electron beam evaporation to a thickness of 20-25 nm as determined by blue interference colors on polished brass. Recently the mineral guys lost their coater so I have also been coating their material to the same blue color they said they used as well. They used resistance heating.
In examining those samples they noticed their EDS/WDS results appearing a little low so we went ahead and coated the standards as well.
More digging has revealed they coat with graphite rods whereas I have been using amorphous C rods. They swear the coats are thicker and looking at theirs-vs-mine it appears so BUT we both evaporated to the same blue color on brass.
Could there be a difference in the optical properties of the graphite and amorphous rods?? I assumed maybe incorrectly that the evaporated material was basically the same from the two rods thus blue brass would also equal the same thickness. Thanks
Scott Whittaker Lab Manager Laboratories of Analytical Biology Smithsonian Institution National Museum of Natural History PO Box 37012 MRC104 Washington DC 20013-7012 202-357-1651
I hope someone knows the answer to this question because I've wondered the same thing... carbon rods vs graphite. another scott
************************************************ ....amphiboles do violence to history... T. Feininger, 2001. (taken out of context) ****************************
Dr. Scott Kuehner kuehner-at-u.washington.edu Dept. of Geological Sciences ph.206-543-8393 Box 351310 Fax 206-616-6873 The University of Washington Seattle, Washington 98195-1310 ************************************************
On Fri, 4 Oct 2002, Scott Whittaker wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I have been coating samples via electron beam evaporation to a thickness of } 20-25 nm as determined by blue interference colors on polished brass. } Recently the mineral guys lost their coater so I have also been coating } their material to the same blue color they said they used as well. They used } resistance heating. } } In examining those samples they noticed their EDS/WDS results appearing a } little low so we went ahead and coated the standards as well. } } More digging has revealed they coat with graphite rods whereas I have been } using amorphous C rods. They swear the coats are thicker and looking at } theirs-vs-mine it appears so BUT we both evaporated to the same blue color } on brass. } } Could there be a difference in the optical properties of the graphite and } amorphous rods?? I assumed maybe incorrectly that the evaporated material } was basically the same from the two rods thus blue brass would also equal } the same thickness. Thanks } } Scott Whittaker } Lab Manager } Laboratories of Analytical Biology } Smithsonian Institution } National Museum of Natural History } PO Box 37012 MRC104 } Washington DC 20013-7012 } 202-357-1651 } } }
Does anybody know where I can buy a book called "Secondary Ion Mass Spectrometry: Basic concepts, Instrumental aspects, applications and trends " by Benninghoven, A.; F.G.Rudenauer F.G., and Werner H.W.
I used to use carbon/graphite rods for making thin carbon film. I do find that using thermal evaporation, the properties of the film originated from graphite or "carbon" are different. It looks like, "graphite" films are more fragile, less stable and somehow more "crumbly". I do notice that "graphite" evaporates at lower current (thermal) also. This difference (in properties of the film) is less pronounced when Electron Beam Evaporator (Electron Gun) was used... But, still, I prefer to use spectrographic quality "carbon rods".
In general, carbon thickness is difficult to determine precisely. I am using Maxtec TM-450(?) thickness monitor with preciseness 0.01 nm. Even with this instrument, the real thickness may vary up to 10-15% (never measured exact value, but have "feelings"). Sergey
At 08:00 AM 10/4/02 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sergey Ryazantsev, Ph.D. Electron Microscopy Department of Biological Chemistry, School of Medicine University of California, Los Angeles Box 951737 Los Angeles, CA 90095-1737
University of Connecticut Institute of Materials Science
Postdoctoral Position Displacive Phase Transformations
A post-doctoral position is available immediately within the Institute of Materials Science at the University of Connecticut to work on displacive phase transformations in crystalline solids. The systems to be studied include metallic pseudoelastic alloys and ferroelectric thin films. The ideal candidate will have experience in both microstructural characterization (TEM, SEM etc.) and phenomenological/crystallographic modeling of phase transformations. This position is a one-year appointment, with possible renewal for a second year.
To apply, please send a complete resume, together with a list of publications and contact details for 3 referees to either Prof. S. Pamir Alpay (p.alpay-at-ims.uconn.edu) or Prof. M. Aindow (m.aindow-at-uconn.edu). Screening of applications will begin immediately, and will continue until the position is filled. We encourage applications from under-represented groups, including minorities, women and people with disabilities.
Mark Aindow, Associate Professor, Department of Metallurgy and Materials Engineering Institute of Materials Science, 97 North Eagleville Road, Unit 3136 University of Connecticut, Storrs, CT 06269-3136, USA
FIRST CALL FOR PRESENTATIONS & FALL MEETING ANNOUNCEMENT
SOCIETY: Central States Microscopy and Microanalysis Society (CSMMS)
DATE: November 15, 2002. From 9 AM to 4 PM.
LOCATION: Giant City State Park. Carbondale, Illinois
DESCRIPTION:
The CSMMS will hold its Fall meeting at the rustic lodge at the Giant City State Park in Carbondale, Illinois. Located in a lovely wooded park, the lodge will serve as the focal point for the presentations and luncheon banquet. The registration and orientation will begin at 9 AM and presentations will commence at 9:30 AM.
The first half-day will involve research presentations, followed by a luncheon and business meeting. The second half-day will involve technical presentations, demonstrations and discussions. For example, we will have presentations on digital imaging (Production of Large Format Posters, Digital Publishing, etc.) as well as discussions of useful techniques and technical pointers. Topics may cover any type of microscopy and microanalysis.
If you are interested in attending or presenting, please contact me for details.
############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu ##############################################################
In specification it indicates 0.01 nm accuracy as far as I remember, but as I mentioned in my original posting, it seems to me that I do have approximately 10% difference in the film thickness even with that smart thickness monitor. In general, I am happy with that instrument. It's very solid and sensitive enough in 10-20 A scale. Sergey
At 04:15 PM 10/4/02, you wrote: } Dear Sergey, } Does your thickness monitor really measure down to 0.01 nm } accuracy?? That's amazing. } } Tobias } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
I am JOHNSON MULETE JR. the son of Mr. Luke Radebe MULETE from Zimbabwe. I am sorry this mail Will Surprise you, though we do not know, I got your contact Through South African Chamber of Commerce.
During the current war against white farmers in Zimbabwe and the support of President Robert Mugabe to Claim all the white owned farms in our country to gain Favor for re-election. All white farmers were asked to Surrender their farms to the government for Re-distribution and infact to his political party Members and my father though black, was the treasury Of the farmers association and a strong member of an Opposition party that did not support the presidents Idea. He then ordered his party members and the police Under his pay role to invade my fathers farm and burn Down everything in the farm. They killed my Father and took away a lot of items from his farm.
After the death of my father, our local pastor and a Close friend of my father handed us over will Documents with instructions from my father that we Should leave Zimbabwe for South Africa incase any Thing happen to him, process the release of the fund Kept in custody for us in a security company unknown To the company that the content is money hence it was deposited as Personal Belongings and Ensure that we do not remain there as we could Easily be found by his enemies. The total amount is US$21.5M.We are therefore soliciting for Your assistance to help us move the fund out of South Africa, as our fate and future is far from Reality, Hence this mail to you. The president's present ban of International Press into Zimbabwe is just a few of the Unthinkable things he is committing in my Country.
I have tried to reach my father's close friend Mr. John Casabas from Australia also a farmer who was Leaving in Zimbabwe with us but left with his family Late last year following this ugly development to no Avail.
Should you be interested to help us, contact me Immediately via email for easy communication and I Will furnish you with the time frame and modalities of The transaction. Please note that this transaction is} 100% confidential and risk free and will not endanger You or us in any way. We have resolved to give you 20% Of the total sum upon confirmation of the fund in any Account of your choice were the incident of taxation Will not take much tool on the money and we look Forward to coming over to your country to invest our Share and settle there. I will provide you my phone number As soon as you indicate your willingness to assist us so that our conversation can be 100%confidential.
Please do not use the reply button, reply only to(johnmulte-at-netscape.net} Please take note.
ANYBODY perfected the SEM "shallow and deep(bottom and no bottom-black) hole area(morphometry) size analysis?? I'm still having trouble with thresholding, flatness, edge defintion and mostly repeatability and accuracy. +_20-50% isn't of much use. Manual edge detection is too slow and not much more repeatable. I'd like to see at least +_5%. jeffrey day/mesqite(o), texas
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Cle02JS12-at-aol.com) from on Saturday, October 5, 2002 at 01:33:09 ---------------------------------------------------------------------------
Email: Cle02JS12-at-aol.com Name: Sandy
Education: 9-12th Grade High School
Location: Washington State
Question: Need referral on how/where to find replacement part for an older college level microscope. Emblem indicates "Alpha Omega-Spencer". Under side indicates American Optical Co.
I am not quite sure what the problem is that you are referring to, but you may want to check out our ACT module (check our web site). It is designed to find edges and do geometric measurements on those edges, using tools developed for the semiconductor industry. If you want to send me an image for a trial run, please send it to my address below.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: charles j day [mailto:wa5ekh-at-juno.com] Sent: Friday, October 04, 2002 8:03 AM To: DrJohnRuss-at-aol.com-at-sparc5.microscopy.com Cc: microscopy-at-sparc5.microscopy.com; NIH-IMAGE-at-LIST.NIH.GOV
ANYBODY perfected the SEM "shallow and deep(bottom and no bottom-black) hole area(morphometry) size analysis?? I'm still having trouble with thresholding, flatness, edge defintion and mostly repeatability and accuracy. +_20-50% isn't of much use. Manual edge detection is too slow and not much more repeatable. I'd like to see at least +_5%. jeffrey day/mesqite(o), texas
try "www.google.com" enter "Benninghoven Rudenauer" brings up this URL: http://www.addall.com/Browse/Detail/0471010561.html states:
Secondary Ion Mass Spectrometry Basic Concepts, Instrumental Aspects, Applications & Trends A. Benninghoven , H. W. Werner , F. G. Rudenauer Binding: Hardcover, 1 edition, 1264 pages Publisher: Wiley, John and Sons, Incorporated Published Date: 01/01/1987 List: USD $395.00 ISBN: 0471010561
} From Microscopy-request-at-sparc5.microscopy.com Fri Oct 4 20:06:03 2002 } Subject: A SIMS book } Date: Fri, 4 Oct 2002 17:29:21 +0200 } From: "Jiao, Chengge" {chengge.jiao-at-uk.feico.com} } To: "Microscopy-at-MSA.Microscopy.Com" {Microscopy-at-sparc5.microscopy.com} } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear All, } } Does anybody know where I can buy a book called } "Secondary Ion Mass Spectrometry: Basic concepts, } Instrumental aspects, applications and trends " } by Benninghoven, A.; F.G.Rudenauer F.G., and Werner H.W. } } Thanks for your information. } } Chengge Jiao }
There have been several postings on the subject of (real) carbon vs. graphite rods for use in carbon coating in a vacuum evaporator.
In many instances, I find that our customers are purchasing "carbon" rods but which are really graphite rods. I am not suggesting that those who offer the product as being truly a carbon rod are doing something wrong because it seems like when we use the words "carbon rods" we don't really think of it as being carbon or graphite but somehow we know what we all mean when we say we use "carbon rods".
But graphite (when used as a "carbon rod") is described in terms of its porosity; indeed, there is a wide range of densities of graphite rods. It is our perception, but not as a result of exhaustive testing, that the higher the porosity ( and lower the density), when one evaporates, there is increased "sparking". I think I would be safe in saying that the "conventional wisdom" suggests that this is undesirable. At the wholesale level (e.g. the large quantities that someone like SPI Supplies purchases) there is in fact a substantial difference in the price between graphite rods of higher porosity vs. lower porosity. We believe that the lower porosity (and accompanying higher density) results in less sparking, perhaps less contaminating nanotubes (which are a bi-product of this kind of evaporation) and other artifactual features.
Actual "carbon" rods have the lowest porosity. Although we have not tested it to the point that I would stake my life on it, real carbon rods seem to spark a whole lot less (if at all) and the final film seems to be much more featureless, but we think that is due to the reduced amount of "sparking". Further information can be found at URL http://www.2spi.com/catalog/spec_prep/carbon-graphite-rods.html
I would be most appreciative of any comments pro or con relative to this analysis. I have been operating on the assumption that this is more or less correct. If there are other points of view, I would be the first to want to know what they are.
Disclaimer: SPI Supplies has offered for some time both carbon rods as carbon rods and graphite rods as graphite rods.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
----------------------------------------------------------------------- The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
I used to use carbon/graphite rods for making thin carbon film. I do find that using thermal evaporation, the properties of the film originated from graphite or "carbon" are different. It looks like, "graphite" films are more fragile, less stable and somehow more "crumbly". I do notice that "graphite" evaporates at lower current (thermal) also. This difference (in properties of the film) is less pronounced when Electron Beam Evaporator (Electron Gun) was used... But, still, I prefer to use spectrographic quality "carbon rods".
In general, carbon thickness is difficult to determine precisely. I am using Maxtec TM-450(?) thickness monitor with preciseness 0.01 nm. Even with this instrument, the real thickness may vary up to 10-15% (never measured exact value, but have "feelings"). Sergey
At 08:00 AM 10/4/02 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sergey Ryazantsev, Ph.D. Electron Microscopy Department of Biological Chemistry, School of Medicine University of California, Los Angeles Box 951737 Los Angeles, CA 90095-1737
ANYBODY perfected the SEM "shallow and deep(bottom and no bottom-black) hole area(morphometry) size analysis?? I'm still having trouble with thresholding, flatness, edge defintion and mostly repeatability and accuracy., etc. +_20-50% isn't of much use. Manual edge detection is too slow and not much more repeatable. I'd like to see at least +_5%. jeffrey day/mesqite(o), texas
Graphite and carbon rods I have (from Ted Pella, I believe) are easily distinguished: graphite is grayish and carbon is deep black. There is also difference in hardness: carbon is harder. I really like your point that graphite produce more "features" when evaporated. It support my intuitional decision to use "carbon". It seems to me it produces more uniform film... Thanks. Sergey
At 10:06 AM 10/6/02 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } } "crumbly". I do notice that "graphite" evaporates at lower current } } } (thermal) also. This difference (in properties of the film) is less } } } pronounced when Electron Beam Evaporator (Electron Gun) was used... } } } But, still, I prefer to use spectrographic quality "carbon rods". } } }
That's funny. When I inadvertantly changed over from 'carbon' to 'graphite, I found that the graphite needed so much current that the rod holder on my Edwards 306 became very hot. I couldn't easily get sufficient evaporation, even at maximum current, so I switched back to carbon.
cheers
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
You may have seen this business before and ignored it. I know I did - many times! However, please take a few moments to read this letter. I was amazed when the profit potential of this business finally sunk in... and it works!
With easy-to-use e-mail tools and opt-in e-mail, success in this business is now fast, easy and well within the capabilities of ordinary people who know little about internet marketing. And the earnings potential is truly staggering!
I'll make you a promise. READ THIS E-MAIL TO THE END! - follow what it says to the letter - and you will not worry whether a RECESSION is coming or not, who is President, or whether you keep your current job or not. Yes, I know what you are thinking. I never responded to one of these before either. One day though, something just said: You throw away $25.00 going to a movie for 2 hours with your wife. What the heck. Believe me, no matter where you believe those feelings come from, I thank every day that I had that feeling.
I cannot imagine where I would be or what I would be doing had I not. Read on. It's true. Every word of it. It is legal. I checked. Simply because you are buying and selling something of value.
AS SEEN ON NATIONAL TV:
Making over half a million dollars every 4 to 5 months from your home.
THANKS TO THE COMPUTER AGE AND THE INTERNET! ================================================== BE AN INTERNET MILLIONAIRE LIKE OTHERS WITHIN A YEAR!!!
Before you say Bull, please read the following. This is the letter you have been hearing about on the news lately. Due to the popularity of this letter on the internet, a national weekly news program recently devoted an entire show to the investigation of this program described below, to see if it really can make people money. The show also investigated whether or not the program was legal.
Their findings proved once and for all that there are absolutely NO laws prohibiting the participation in the program and if people can 'follow the simple instructions' they are bound to make some mega bucks with only $25 out of pocket cost.
DUE TO THE RECENT INCREASE OF POPULARITY & RESPECT THIS PROGRAM HAS ATTAINED, IT IS CURRENTLY WORKING BETTER THAN EVER.
This is what one had to say: Thanks to this profitable opportunity. I was approached many times before but each time I passed on it. I am so glad I finally joined just to see what one could expect in return for the minimal effort and money required. To my astonishment, I received a total $610,470.00 in 21 weeks, with money still coming in.
Pam Hedland, Fort Lee, New Jersey. ================================================== Another said: This program has been around for a long time but I never believed in it. But one day when I received this again in the mail I decided to gamble my $25 on it. I followed the simple instructions and walaa ..... 3 weeks later the money started to come in. First month I only made $240.00 but the next 2 months after that I made a total of $290,000.00. So far, in the past 8 months by re-entering the program, I have made over $710,000.00 and I am playing it again. The key to success in this program is to follow the simple steps and NOT change anything.
More testimonials later but first, =======
==== PRINT THIS NOW FOR YOUR FUTURE REFERENCE ==== $$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$
If you would like to make at least $500,000 every 4 to 5 months easily and comfortably, please read the following...THEN READ IT AGAIN and AGAIN!!!
FOLLOW THE SIMPLE INSTRUCTION BELOW AND YOUR FINANCIAL DREAMS WILL COME TRUE, GUARANTEED!
INSTRUCTIONS:
=====Order all 5 reports shown on the list below =====
For each report, send $5 CASH, THE NAME & NUMBER OF THE REPORT YOU ARE ORDERING and YOUR E-MAIL ADDRESS to the person whose name appears ON THAT LIST next to the report. MAKE SURE YOUR RETURN ADDRESS IS ON YOUR ENVELOPE TOP LEFT CORNER in case of any mail problems.
===WHEN YOU PLACE YOUR ORDER, MAKE SURE === ===YOU ORDER EACH OF THE 5 REPORTS! === You will need all 5 reports so that you can save them on your computer and resell them. YOUR TOTAL COST $5 X 5 = $25.00.
Within a few days you will receive, via e-mail, each of the 5 reports from these 5 different individuals. Save them on your computer so they will be accessible for you to send to the 1,000's of people who will order them from you. Also make a floppy of these reports and keep it on your desk in case something happens to your computer.
IMPORTANT - DO NOT alter the names of the people who are listed next to each report, or their sequence on the list, in any way other than what is instructed below in steps 1 through 6 or you will lose out on the majority of your profits. Once you understand the way this works, you will also see how it will not work if you change it.
Remember, this method has been tested, and if you alter it, it will NOT work!!! People have tried to put their friends'/relatives' names on all five thinking they could get all the money. But it does not work this way. Believe us, some have tried to be greedy and then nothing happened. So Do Not try to change anything other than what is instructed. Because if you do, it will not work for you. Remember, honesty reaps the reward!!!
This IS a legitimate BUSINESS. You are offering a product for sale and getting paid for it. Treat it as such and you will be VERY profitable in a short period of time.
1.. After you have ordered all 5 reports, take this advertisement and REMOVE the name & address of the person in REPORT # 5. This person has made it through the cycle and is no doubt counting their fortune.
2.. Move the name & address in REPORT # 4 down TO REPORT # 5.
3.. Move the name & address in REPORT # 3 down TO REPORT # 4.
4.. Move the name & address in REPORT # 2 down TO REPORT # 3.
5.. Move the name & address in REPORT # 1 down TO REPORT # 2
6.... Insert YOUR name & address in the REPORT # 1 Position.
PLEASE MAKE SURE you copy every name & address ACCURATELY! This is critical to YOUR success.
================================================== **** Take this entire letter, with the modified list of names, and save it on your computer. DO NOT MAKE ANY OTHER CHANGES.
Save this on a disk as well just in case you lose any data. To assist you with marketing your business on the internet, the 5 reports you purchase will provide you with invaluable marketing information which includes how to send bulk e-mails legally, where to find thousands of free classified ads and much more. There are 2 primary methods to get this venture going:
METHOD # 1: BY SENDING BULK E-MAIL LEGALLY ==================================================
Let's say that you decide to start small, just to see how it goes, and we will assume you and those involved send out only 5,000 e-mails each. Let's also assume that the mailing receives only a 0.2% (2/10 of 1%) response (the response could be much better but let's just say it is only 0.2%). Also many people will send out hundreds of thousands of e-mails instead of only 5,000 each.
Continuing with this example, you send out only 5,000 e-mails. With a 0.2% response, that is only 10 orders for report # 1. Those 10 people responded by sending out 5,000 e-mails each for a total of 50,000. Out of those 50,000 e-mails only 0.2% responded with orders. That's 100 people who responded and ordered Report # 2.
Those 100 people mail out 5,000 e-mails each for a total of 500,000 e-mails. The 0.2% response to that is 1000 orders for Report # 3.
Those 1000 people send 5,000 e-mails each for a total of 5 million e-mails sent out. The 0.2% response is 10,000 orders for Report # 4.
Those 10,000 people send out 5,000 e-mails each for a total of 50,000,000 (50 million) e-mails. The 0.2% response to that is 100,000 orders for Report # 5.
THAT'S 100,000 ORDERS TIMES $5 EACH = $500,000.00 (half a million dollars).
Your total income in this example is: 1..... $50 + 2..... $500 + 3.....$5,000 + 4..... $50,000 + 5.... $500,000 .... Grand Total=$555,550.00
NUMBERS DO NOT LIE. GET A PENCIL & PAPER AND FIGURE OUT THE WORST POSSIBLE RESPONSES AND NO MATTER HOW YOU CALCULATE IT, YOU WILL STILL MAKE A LOT OF MONEY! ==================================================
REMEMBER FRIEND, THIS IS ASSUMING ONLY 10 PEOPLE ORDERING OUT OF 5,000 YOU MAILED TO. Dare to think for a moment what would happen if everyone or half or even one 4th of those people mailed 100,000 e-mails each or more?
There are over 150 million people on the internet worldwide and counting, with thousands more coming online every day. Believe me, many people will do just that, and more!
METHOD # 2: BY PLACING FREE ADS ON THE INTERNET ==================================================
Advertising on the net is very, very inexpensive and there are hundreds of FREE places to advertise. Placing a lot of free ads on the internet will easily get a larger response. We strongly suggest you start with Method # 1 and add METHOD # 2 as you go along. For every $5 you receive, all you must do is e-mail them the report they ordered. That's it. Always provide same day service on all orders.
This will guarantee that the e-mail they send out, with your name and address on it, will be prompt because they cannot advertise until they receive the report.
===========AVAILABLE REPORTS ==================== The reason for the cash is not because this is illegal or somehow wrong. It is simply about time. Time for checks or credit cards to be cleared or approved, etc. Concealing it is simply so no one can SEE there is money in the envelope and steal it before it gets to you.
ORDER EACH REPORT BY ITS NUMBER & NAME ONLY. Notes: Always send $5 cash (U.S. CURRENCY or EURO) for each report. Checks NOT accepted. Make sure the cash is concealed by wrapping it in at least 2 sheets of paper. On one of those sheets of paper, write the NUMBER & the NAME of the report you are ordering, YOUR E-MAIL ADDRESS and your name and postal address.
PLACE YOUR ORDER FOR THESE REPORTS NOW : ================================================== REPORT # 1: The Insider's Guide To Advertising for Free On The Net Order Report #1 from:
EARL PACE P.O. BOX 371 SUCC. ST-MARTIN LAVAL H7V 3P6 CANADA ______________________________________________________ REPORT # 2: The Insider's Guide To Sending Bulk Email On The Net Order Report # 2 from:
REPORT # 4: How To Become A Millionaire Using MLM & The Net Order Report # 4 from:
Jean Lafortune 107, rue Lauriston 75116 Paris FRANCE _______________________________________________________
REPORT # 5: How To Send Out One Million Emails For Free Order Report # 5 From:
Stuart Iles 9 Elm Way Hartshill NUNEATON CV10 0XS United Kingdom
______________________________________________________ $$$$$$$$$ YOUR SUCCESS GUIDELINES $$$$$$$$$$$
Follow these guidelines to guarantee your success:
=== If you do not receive at least 10 orders for Report #1 within 2 weeks, continue sending e-mails until you do.
=== After you have received 10 orders, 2 to 3 weeks after that you should receive 100 orders or more for REPORT # 2. If you do not, continue advertising or sending e-mails until you do.
** Once you have received 100 or more orders for Report # 2, YOU CAN RELAX, because the system is already working for you, and the cash will continue to roll in ! THIS IS IMPORTANT TO REMEMBER: Every time your name is moved down on the list, you are placed in front of a different report.
You can KEEP TRACK of your PROGRESS by watching which report people are ordering from you. IF YOU WANT TO GENERATE MORE INCOME SEND ANOTHER BATCH OF E-MAILS AND START THE WHOLE PROCESS AGAIN. There is NO LIMIT to the income you can generate from this business !!! ================================================= FOLLOWING IS A NOTE FROM THE ORIGINATOR OF THIS PROGRAM: You have just received information that can give you financial freedom for the rest of your life, with NO RISK and JUST A LITTLE BIT OF EFFORT. You can make more money in the next few weeks and months than you have ever imagined. Follow the program EXACTLY AS INSTRUCTED. Do Not change it in any way. It works exceedingly well as it is now.
Remember to e-mail a copy of this exciting report after you have put your name and address in Report #1 and moved others to #2 .....# 5 as instructed above. One of the people you send this to may send out 100,000 or more e-mails and your name will be on every one of them.
Remember though, the more you send out the more potential customers you will reach. So my friend, I have given you the ideas, information, materials and opportunity to become financially independent.
IT IS UP TO YOU NOW !
=============MORE TESTIMONIALS=============== My name is Mitchell. My wife, Jody and I live in Chicago. I am an accountant with a major U.S. Corporation and I make pretty good money. When I received this program I grumbled to Jody about receiving 'junk mail'. I made fun of the whole thing, spouting my knowledge of the population and percentages involved. I 'knew' it wouldn't work. Jody totally ignored my supposed intelligence and few days later she jumped in with both feet. I made merciless fun of her, and was ready to lay the old 'I told you so' on her when the thing didn't work.
Well, the laugh was on me! Within 3 weeks she had received 50 responses. Within the next 45 days she had received total $ 147,200.00 ......... all cash! I was shocked. I have joined Jody in her 'hobby'.
Mitchell Wolf M.D., Chicago, Illinois ================================================ Not being the gambling type, it took me several weeks to make up my mind to participate in this plan. But conservative as I am, I decided that the initial investment was so little that there was just no way that I wouldn't get enough orders to at least get my money back. I was surprised when I found my medium size post office box crammed with orders. I made $319,210.00 in the first 12 weeks.
The nice thing about this deal is that it does not matter where people live. There simply isn't a better investment with a faster return and so big.
Dan Sondstrom, Alberta, Canada ================================================= I had received this program before. I deleted it, but later I wondered if I should have given it a try. Of course, I had no idea who to contact to get another copy, so I had to wait until I was e-mailed again by someone else......... 11 months passed then it luckily came again...... I did not delete this one! I made more than $490,000 on my first try and all the money came within 22 weeks.
Susan De Suza, New York, N.Y. ================================================= It really is a great opportunity to make relatively easy money with little cost to you. I followed the simple instructions carefully and within 10 days the money started to come in. My first month I made $20, in the 2nd month I made $560.00 and by the end of the third month my total cash count was $362,840.00. Life is beautiful, Thanx to internet.
Fred Dellaca, Westport, New Zealand =================================================
ORDER YOUR REPORTS TODAY AND GET STARTED ON YOUR ROAD TO FINANCIAL FREEDOM !
================================================= If you have any questions of the legality of this program, contact the Office of Associate Director for Marketing Practices, Federal Trade Commission, Bureau of Consumer Protection, Washington, D.C.
This message is sent in compliance of the proposed bill SECTION 301, paragraph (a)(2)(C) of S. 1618.
* This message is not intended for residents in the State of Washington, Virginia or California, screening of addresses has been done to the best of our technical ability.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (arrobert-at-mtu.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, October 6, 2002 at 19:39:15 ---------------------------------------------------------------------------
Email: arrobert-at-mtu.edu Name: Amy Roberts
Organization: Michigan Technological University
Education: Undergraduate College
Location: Houghton, Mi United States of America
Question: I am in search of specimen preparation procedures for transmission electron microscopy for a gram negative and a gram positive bacteria. Could anyone point me to a journal or publication with this information?
I would like to ask if anyone could fax me a copy of the operating instructions for a Sorvell MT2-B Ultra microtome. It is motorized if that matters.
This is for a lady that is not on the list and that works at another research center.
Thank you in advance for your help and time.
724-325-5105 My FAX
Sincerely,
Paul Beauregard Senior Research Associate PPG Industries Monroeville Technical Center Electron Microscopy 440 College Park Drive Monroeville, PA 15146
} To calculate the size of one micron in ANY microscopic photograph, one only has to know the magnification. Take the mag and convert it to KX. That number is the number of millimeters that represents one micrometer on your photo. For example, 81385X = 81.385KX and 1micron is 81.385 mm. 516X = 0.516KX and one micron is .516 mm. 1,300,000X = 1300KX and 1300 mm is 1 Micron or 130MMs is 0.1 microns or 13 mm is 10NMs or 100Å. } } }
} I have been coating samples via electron beam evaporation to a } thickness of } 20-25 nm as determined by blue interference colors on polished brass. } Recently the mineral guys lost their coater so I have also been coating } their material to the same blue color they said they used as } well. They used } resistance heating. } } In examining those samples they noticed their EDS/WDS results appearing a } little low so we went ahead and coated the standards as well. } } More digging has revealed they coat with graphite rods whereas I have been } using amorphous C rods. They swear the coats are thicker and looking at } theirs-vs-mine it appears so BUT we both evaporated to the same blue color } on brass. } } Could there be a difference in the optical properties of the graphite and } amorphous rods??
I can't imagine any difference once the carbon has been evaporated ... i.e., there may be a difference with respect to resistance and temperature of evaporation (sublimation?) between vitreous carbon and graphite, but once the transformation happens, carbon is carbon.
I happen to use the red-to-blue transistion, because it is very distinguishable color change and should result in a coating thickness of 22nm +- 1nm.
cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
*************************************************************** First Announcement
Electron Microscopy and Analysis Conference 2003 The University of Oxford 3 - 5 September 2003
Organised by the Electron Microscopy and Analysis Group of the Institute of Physics Co-sponsored by the Royal Microscopical Society
Scientific Content Electron Microscopy is going through the most revolutionary period since its invention, with leaps in performance of sources, imaging lenses and spectrometers all at roughly the same time. Scanning probe techniques are also maturing into useful analytical tools. This is an ideal time for the microscopy community to appraise when, how and why we use different techniques to gain different scientific insights. The conference will address all aspects of electron and scanning probe microscopy and associated spectroscopy including:
o New Instrumentation - including Aberration Correction and Monochromation o New Information from New Instrumentation - Applications Highlighting Novel Techniques o Advances in Nanoanalysis - EDX, PEELS and Other Analytical Techniques o State-of-the-art SEM, Scanning Probe Microscopy and Surface Science o High-resolution Electron Microscopy and Electron Crystallography o Advances in Sample Preparation
Applications of microscopy and related techniques will be divided into sessions according to demand. These will include experiments in:
o Biogenic Materials o Catalysis o Ceramics and Electroceramics o Composites o Environmental Samples o Ferrous and Non-Ferrous Metals o Fullerenes, Hard Carbon and Related Materials o Intermetallics o Interfaces and Surfaces o Magnetic Materials o Sensor Materials o Semiconductors oSuperconductors
Sessions of oral and poster presentations will be arranged, and, where possible, associated with invited keynote speakers. Papers from post-graduate students are particularly encouraged; student bursaries will be available through the Institute of Physics EMAG Group.
Advanced School The Advanced School provides an opportunity for postgraduate students, research assistants and others with an interest to learn about a range of advanced imaging techniques in electron and related microscopies techniques from experts in the field. The school will involve a combination of tutorial lectures and hands-on demonstrations in the Department of Materials, using the wide range of sophisticated instrumentation available there. The lecturers/demonstrators will be leading exponents in the techniques concerned. There will be considerable opportunity for interaction. To maximise the effectiveness of the School, numbers will be limited to about 20.
Trade Exhibition Accompanying the conference there will be a trade exhibition where delegates and visitors will be able to see and discuss the latest products and developments.
Social Programme The social programme will include a welcome reception, an Exhibitor Buffet and a conference dinner. Further details will be available in due course.
Conference Venue The scientific sessions, the Exhibition and the poster presentations will take place at the Oxford University Examination Schools. For more information about how to get to Oxford please visit the Oxford University web site at http://www.ox.ac.uk/aboutoxford/how.shtml
Important Dates January 2003 all for Papers 21 March 2003 Deadline for Abstract Submission May 2003 Registration Mailing 18 July 2003 Deadline for Registration
Organising Committee
EMAG Chairman R M D Brydson Local Committee Chairman A Petford-Long/J Hutchison Programme Organiser A Bleloch Proceedings Editor S McVitie Exhibition Organiser R Doole Advanced School Organiser D J Cockayne
General Enquiries Jasmina Bolfek-Radovani, The Institute of Physics, 76 Portland Place, London W1B 1NT Tel +44 (0)20 7470 4800 Fax: +44 (0)20 7470 4900 Email: conferences-at-iop.org http://physics.iop.org/IOP/Confs/EMG/
Exhibition Enquiries Ron Doole, EMAG03, Department of Materials, University of Oxford, Parks Road, Oxford., OX1 3PH Tel: + 44 (0)1865 273701 Fax: + 44 (0)1865 283333 Email:ron.doole-at-materials.oxford.ac.uk
Media Registration Accredited journalists are welcome to attend all or part of this meeting free-of-charge. To qualify for free registration please contact Alice Bows, Press Officer, Public Affairs Department, Tel: +44 (0)20 7470 4800, Fax: +44 (0)20 7470 4848, Email: alice.bows-at-iop.org.
In an analysis I did several years ago I wanted to place a controlled amount of carbon on a surface and measure the ultimate strength of a gold to gold weld in a microcircuit. We had great difficulty with this since we had no method to know, with any accuracy, the real deposition thickness. Our target was 100 angstroms, with increments up to 500. We attempted to use Auger, however, with a carbon species and the fact that the first 50 angstroms in the Auger survey are thrown out, we never actually determined what the final result was. Would AFM have been a better choice for this measurement?
Peter Tomic Anadigics, Inc.
-----Original Message----- } From: Sergey Ryazantsev [mailto:sryazant-at-ucla.edu] Sent: Friday, October 04, 2002 9:47 PM To: Microscopy-at-sparc5.microscopy.com
Tobias, good question
In specification it indicates 0.01 nm accuracy as far as I remember, but as I mentioned in my original posting, it seems to me that I do have approximately 10% difference in the film thickness even with that smart thickness monitor. In general, I am happy with that instrument. It's very solid and sensitive enough in 10-20 A scale. Sergey
At 04:15 PM 10/4/02, you wrote: } Dear Sergey, } Does your thickness monitor really measure down to 0.01 nm } accuracy?? That's amazing. } } Tobias } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
7:15pm - Raman Microspectroscopy and Imaging David S. Himmelsbach, United States Department of Agriculture Agricultural Research Service Richard B. Russell Research Center, Athens, Georgia, 30677
8:00pm - Raman Spectroscopy on Isolated Carbon Nanotubes Shin Grace Chou, Center for Materials Science and Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139
Travel: A map and directions are on the Web at http://whereis.mit.edu/bin/map?state=0&pri.x=281&pri.y=123 Parking will be extremely difficult because of on-going road construction on Vassar Street (the main location of on-street parking). If you park in any MIT lot, even in the evening, you will be ticketed unless you have an MIT sticker. There is a public lot on the corner of Vassar St. and Mass. Ave. that may be available. There is a public garage behind the Legal Seafoods Restaurant in Kendall Square (10 mins. walk). There is another garage several blocks North just off Mass. Ave. (2 blocks beyond the NECCO building). It should also be possible to find on-street parking, but be prepared to walk.
If possible, we recommend you use public transport. The number 1 bus from Harvard to Dudley stops on Mass Ave outside the student center. The Red Line stops at Central and Kendall are each 10 minutes walk (use Central if you come from Harvard or Alewife, Kendall if you come from Boston or points South.)
Registration for members is $5.00 and for non-members is $20.00 (this includes a one-year membership in NESM). The deadline for registration is Friday, October 11th. Please contact Paul Bain, NESM Treasurer, (Paul_Bain-at-hms.harvard.edu) if you are planning to attend.
** Anthony J. Garratt-Reed ** MIT Room # 13-1027 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6478 **
We are interested in buying a used 2000FX LN2 and/or heating holder, If anyone would like to sell those used holders please let me know.
Thanks,
Jinguo Wang, Ph.D The Pennsylvania State University Materials Research Institute 194 Materials Research Institute Building University Park, PA 16802 Tel: (814) 865-9285 Fax: (814) 863-8561, (814) 863-0637 email: jqw11-at-psu.edu
We use a great deal of carbon and graphite daily for making our substrates. Some in our lab prefer graphite, some carbon. If you examine the crystalline structure you'll find carbon is more random (less crystalline). Graphite requires more current in the Ladd evaporators. Carbon is more expensive. We offer carbon and high quality graphite. Based on our experience and customer feedback we are of the opinion that excellent results can be attained with carbon or high quality graphite.
John Arnott
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com ----- Original Message ----- } From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} To: "Sergey Ryazantsev" {sryazant-at-ucla.edu} ; {Microscopy-at-sparc5.microscopy.com} Sent: Sunday, October 06, 2002 7:12 PM
Ladies/Gentlemen:
I am searching for TEMs of two fungi; one is our newly discovered Penicillium that is related to several listed below. Need TEM's with good ultrastructure and labeling of internal structures. Will appreciate help by listing references, text books, personal electronmicrographs, etc. Need information for publication purposes.
1) Fusarium verticilloides (moniliforme) or related species.
2) Penicillium species related to new species identified:
a. Penicillium restrictum b. Penicillium dimorphosporum c. Penicillium striatisporum
If TEM's of above Penicillium species not available, then electronmicrographs of monoverticillate, nonvesiculate penicilli with short stipes.
This is off-topic, many apologies, but I am stumped. I am not familiar with mass spectrometers, but have been asked to get information on upgrading a 1070's vintage mass spec - quadrupole. The manufacturer is Nuclide. They want to send the detector output to a PC. Many thanks for any info from vendors or individuals. Nancy
Nancy Piatczyc Bronfman Science Center Williams College Williamstown, MA 01267
Actually, only one direct way to measure REAL thickness I do know: it's embedding in the plastic and cross-sectioning strongly PERPENDICULAR. All other ways are arbitrary. As I mentioned in original report, I do have 10% difference from experiment to experiment for carbon. Heavier elements may be measured, perhaps, more precisely... Sergey
At 07:03 AM 10/7/02, you wrote: } Sergey & Co.; } } In an analysis I did several years ago I wanted to place a controlled amount } of carbon on a surface and measure the ultimate strength of a gold to gold } weld in a microcircuit. We had great difficulty with this since we had no } method to know, with any accuracy, the real deposition thickness. Our } target was 100 angstroms, with increments up to 500. We attempted to use } Auger, however, with a carbon species and the fact that the first 50 } angstroms in the Auger survey are thrown out, we never actually determined } what the final result was. Would AFM have been a better choice for this } measurement? } } Peter Tomic } Anadigics, Inc. } } -----Original Message----- } From: Sergey Ryazantsev [mailto:sryazant-at-ucla.edu] } Sent: Friday, October 04, 2002 9:47 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: SEM Carbon Coating thickness } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Even with a great FESEM as we have, I would be hard pressed to image 100 angstroms of anything with a 10 angstrom accuracy. The FESEM sometimes can do 15 angstroms on an ideal sample on a good day, but I could never resolve 10A/100A. By "perpendicular" x-sectioning, are you referring to mount lap and polish, microtome? Also, what is a "Smart Thickness Monitor" you mention below? Maybe I need one. In short, how does it do the measurement, a stylus like AFM, optical method?
Regards,
Peter
Ps. I would imagine the Los Angeles smog would put 100 angstroms of soot down on a glass slide in short order, unlike pristine New Jersey.
-----Original Message----- } From: Sergey Ryazantsev [mailto:sryazant-at-ucla.edu] Sent: Monday, October 07, 2002 2:54 PM To: Peter Tomic; Microscopy-at-sparc5.microscopy.com
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Ok here's a new twist. It was kinda slow Friday and we ended up running several different rods and simulated samples again coated to blue on brass. My brass washer was the same color as theirs. The consensous upstairs is still that that my coatings are substantially thicker and looking at the samples I agree they are darker.
They evaporated carbon and sometimes graphite via resistance. I use e-beam evaporation, and graphite rods take about 5 minutes, heating the head to cherry in color so is not really an option. Could the different thickness be due to different density of the films? The e-beam evaporated material in theory has a finer more uniform structure with less space, thus the film is more "dense" There would be more carbon in the film per unit thickness so when I coat to the blue 25nm, more carbon is present.
Resistance evaporation is less controlled and there is less carbon in the film structure, but still colors brass blue at 25nm.
The other option is that a denser film optically reflects differently and the blue is something like 40nm or so. I had ruled that out since I have a thickness monitor and it is reading correctly.
Anyway, just some more thoughts to toss around. I never really paid any attention to the carbon films and have already learned much from the discussion so far. Thanks
What is the actual molecular structure of an "amorphous" carbon rod? Is it really amorphous? Microcrystalline? Something else? Given the crystal form of graphite, I suspect that it is a much more efficient generator of bucky balls, nanotubes, and suchlike structures. This could explain the problems with film thickness and consistency from graphite evaporation vs (amorphous) carbon rods. How would the bucky balls, nanotubes, etc. affect the electrical properties of a carbon film, and thus its behavior in an electron beam? Or is this all wrong and irrelevant?
Has anyone done any AFM/STM on carbon films deposited by these different methods?
Phil
} Scott writes ... } } } I have been coating samples via electron beam evaporation to a } } thickness of } } 20-25 nm as determined by blue interference colors on polished brass. } } Recently the mineral guys lost their coater so I have also been coating } } their material to the same blue color they said they used as } } well. They used } } resistance heating. } } } } In examining those samples they noticed their EDS/WDS results appearing a } } little low so we went ahead and coated the standards as well. } } } } More digging has revealed they coat with graphite rods whereas I have been } } using amorphous C rods. They swear the coats are thicker and looking at } } theirs-vs-mine it appears so BUT we both evaporated to the same blue color } } on brass. } } } } Could there be a difference in the optical properties of the graphite and } } amorphous rods?? } } I can't imagine any difference once the carbon has been evaporated ... } i.e., there may be a difference with respect to resistance and temperature } of evaporation (sublimation?) between vitreous carbon and graphite, but once } the transformation happens, carbon is carbon. } } I happen to use the red-to-blue transistion, because it is very } distinguishable color change and should result in a coating thickness of } 22nm +- 1nm. } } cheerios ... shAf :o) } Avalon Peninsula, Newfoundland } www.micro-investigations.com (in progress)
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
When carbon (or any other material) evaporated (sublimated for carbon?) as I understand, it's not mono-atomic "vapor". It contains atomic clusters of different size. Those clusters may be produced during "evaporation" or later in the "vapor" clouds. I could imagine that evaporation conditions, the density of cloud etc may shift local conditions in the favor of some carbon state. Condensation conditions on the surface may also vary (temperature etc). So, to me it's easy to imagine how it happens that carbon film is heterogeneous (mica steps could initiate some specific condensation). I was thinking to do AFM on different "carbons". It should be done in the vacuum environment I guess. Unfortunately, I don't have access to such instrumentation. Sergey
At 01:21 PM 10/7/02, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
I have been told that the problem we are currently experiencing with our ~ 1980 JEOL 100CX has been seen before.
We would very much appreciate any information or advice.
We are getting an accumulation of oil mist (DP oil ?) on the inside of the viewing chamber glass in the specimen chamber of our 100CX. Where is it coming from, where do I start looking for the fault ? We no sooner clean it off than it is back again.
Look forward to hearing from listers.
Tony
Tony Bruton Head, Centre for Electron Microscopy University of Natal, Pietermaritzburg Tel +27 (0) 33 260 5155 Fax +27 (0) 33 260 5776 Mob. 082 782 9640 website: www.nu.ac.za/microscopy.asp Email: bruton-at-nu.ac.za Postal address; Private Bag X01, Scottsville, 3209 KwaZulu-Natal South Africa
Excuse me if this is common knowledge... Thirty years ago, the organization I worked for was fabricating carbon "bottles" to contain cadmium sulphide to be heated for vacuum deposition. We ran into the problem that commercial "graphite" contains several percent boron, put in to make the stuff machinable. Since we didn't want boron involved in our process, we had to specify "nuclear-grade" graphite, intended for reactor parts where they didn't want any boron, either. This turned out to be hard enough to get that we laid in a ten-year supply when we finally found some. Are "graphite" evaporator rods boron-free? If not, could that be responsible for the differences reported? I assume amorphous carbon is pure.
Robert Wieland wieland-at-me.udel.edu The very concept of human governance is a moral dilemma: If the people are good, it is a mistake to create authorities over them; If they are not good, it is a mistake to create authorities out of them.
You can easily peg people by listening to them recount a hand: winners talk about how they played the cards they were dealt; non-winners, about how they should have played the cards they were dealt; and losers, about the cards they should have been dealt.
R F Wieland wieland-at-me.udel.edu 39.68N 75.74W Icom R75 Heathkit GR-81 Inverted-L in the attic
Dear Listers, We are looking for a double tilt (tilt-tilt) holder for a Jeol 100C/100CX to examine metallurgical specimens. If you have one available, please reply to me directly (kzeisler-at-wpi.edu).
Thank you very much. Kate Zeisler-Mashl
----------------------------------------- Kathryn L. Zeisler-Mashl, Ph.D. Research Assistant Professor Materials Science and Engineering Program Worcester Polytechnic Institute 100 Institute Road, Worcester, MA 01609 T: (508)831-6025 F: (508)831-5178 kzeisler-at-wpi.edu
Hi Listservers! I have a few questions that I would like to ask you If you feel you know the answer please reply. My university together with local union reclassified staff positions according to Aiken plan. Needless to say my position was downgraded. My questions are: 1. Can EM supervisory position be compared to any on campus staff positions (university has Department of Sciences and Veterinary Medecine)? I supervise small multi--user lab, we have 2 TEM, and we do research, diagnostics, teaching (under- and graduate level) in the field of biological sciences. No fancy staff like X-ray analysis or cryo. 2. What in your opinion is a necessary minimum experience required to perform this job ? I am talking about accepting the position and immediately getting into the stream of work. 3. What in your opinion is the minimum level of education that is required to perform this job?
You can reply directly to me. Thanks in advance Frustrated Dorota
Boron is in the manufacturing process. It's probably in the PPB range in the graphite rods. Some graphite rods may be boron free but you'd have to assume there is some boron in most graphite rods.
Disclaimer: Ladd sells a wide variety of electron microscope related products.
John Arnott
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com ----- Original Message ----- } From: "Robert Wieland" {wieland-at-ME.UDel.Edu} To: "John Arnott" {ladres-at-worldnet.att.net} Cc: "Microscopy Listserver" {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, October 08, 2002 8:38 AM
I am scheduled to attend McCrone's applied polarized light microscopy course later this month. I have a medical background consisting of gross human anatomy and physiology, a limited science background, law enforcement and legal experience, but I currently do not work in a science field. A friend of mine recently provided me with some "hands-on" with a polarized light microscope.
How else can I prepare for this course of instruction? I am paying for this training myself, and I want to get the most out of it.
More on the discussion on carbon/graphite rods used in EM labs for evaporation:
To understand what the situation really is, you have to go back to the "starting" material, which differs between the different manufacturers of the final product. The starting material could be either coke, coal, or what is called "pitch". Since this can come from different geographical locations, the level of impurities varies, including the level of boron which is everywhere.
The first step is the converstion of the pitch to carbon and then the second step is the conversion of the carbon to graphite. At least that is the process used in the production of the SPI Supplies offered rods. The problem arises because boron very easily forms a carbide and that carbide is very difficult to purify out of the final product.
In years gone by, and before analytical instrumentation had the low detection limits of today's instrumentation, it was indeed common to talk about carbon or graphite as being "boron free". Of course, nothing is ever zero, we can only talk about detection limits and say that if present, it is less than some amount.
The instrumentation used for the control of boron in the SPI Supplies carbon and graphite rod products has a detection limit of slightly more than 1 ppm and we keep our product to a boron level of less than 2 ppm.
The purification cost is a major cost element of the delivery of high purity carbon or graphite rods; not all applications require this kind of purity. The way products are presented, either in catalogs or websites, might suggest they are all the same but they are not all the same. I can not address the purity of the carbon or graphite rods sold by others, only that offered by SPI Supplies. But certainly this level of purity of boron is well beyond that needed by anyone using these products in an EM lab application.
} From our experience, the biggest reason why different rods from different sources evaporate differently is due to differences in the density of the grahpite rods.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
Boron is in the manufacturing process. It's probably in the PPB range in the graphite rods. Some graphite rods may be boron free but you'd have to assume there is some boron in most graphite rods.
Disclaimer: Ladd sells a wide variety of electron microscope related products.
John Arnott
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com ----- Original Message ----- } From: "Robert Wieland" {wieland-at-ME.UDel.Edu} To: "John Arnott" {ladres-at-worldnet.att.net} Cc: "Microscopy Listserver" {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, October 08, 2002 8:38 AM
Hi group - anybody out there that currently uses a carbon coater with a turbo pump? Looks like our Pelco carbon coater died and parts are hard to find. Can any of you recommend a carbon coater that you are happy with? Thanks Barb
Hi didn't really get a response from my last query - anybody currently using a CL with SEM? We are looking to upgrade our system and get a cold stage. Thanks Barb
This problem might be caused by several conditions. First, determine what oil is present in the projection chamber. Possibilities are: Santovac (DP), Hydrocarbon (PVP), and air compressor oil, which could be either Hydrocarbon or an Ester, depending on compressor type. The latter might be irrelevant in case that you have replaced original JEOL compressor with a dry compressor, however: an old compressor oil could be present in the pneumatic lines/valves for years, and might eventually show up on the vacuum side of pneumatic valves upon moveable seal(s) failure.
I use a highly volatile solvent (such as Pentane), others may work too. Pentane dissolves Hydrocarbon oil very well and fast, Santovac very poorly, and Ester oil to some degree. In addition, compressor oil (especially one sitting in the lines and valves for years, and especially Ester oil) has deep red/brown color, clearly visible even in microscopic droplets.
Remove a small polished metal part with oil mist visible on it from the contaminated area. Observe oil mist under stereo microscope, see the color, try consistency of the droplets with the needle. Santovac is much more viscous, especially in a cool room. Rinse that part briefly in Pentane, see if some droplets have disappeared, or were reduced in size more than others.
Many scenarios are possible, including following: a) ODP(s) cooling failed (clogged water lines) - Santovac in the column/viewing chamber. Likely scenario.
b) Pneumatically actuated vacuum valves acting slowly (damage, dirt, deformed moving seals, compressor oil and water in pneumatic cylinders)- PVP Hydrocarbon oil or/and Santovac are blown into vacuum when valves switching for venting or rough pumping is delayed. Likely scenario.
c) Damaged seals in the pneumatically actuated vacuum valves- compressor oil in the column/viewing chamber. BTW, do you drain oil and water from air compressor tank regularly? Do you have water mist filter installed between compressor and a TEM? Likely scenario.
d) Damaged PVP- Hydrocarbon oil makes it's way into the ODP, and from there into column/viewing chamber. Unlikely scenario.
e) Pirani gauges or circuits are defective- vacuum sequences are out of order. Connect stand-alone vacuum gauge to a TEM (multiple ports) and verify switching points/thresholds. Unlikely scenario.
Unfortunately, an easy fix is unlikely for a problem like this. I recommend spending some time and conduct an investigation, with vacuum system diagram in mind. This will help to trace problem more accurately, and avoid unnecessary disassembly.
Possible remedies may include cleaning/re-packing pneumatically actuated vacuum valves, cleaning pneumatic lines, cleaning an ODP and replacing the Santovac, replacing worn out air compressor, replacing/servicing PVP.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax (678)467-0012 mobile
This message is made of 100% recycled electrons.
This address can not receive messages larger than 15 kb without prior notification.
----- Original Message ----- } From: Tony Bruton {Bruton-at-nu.ac.za} To: {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, October 08, 2002 8:04 AM
There is an excellent book on the subject by F. Donald Bloss called "Optical Crystallography" and it is available from the Minerological Society of america for about $25.00.
Don ----- Original Message ----- } From: "Drew Price" {dprice2-at-sigecom.net} To: {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, October 08, 2002 12:00 PM
Why not ask the people who run the course?
Geoff
Drew Price wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I am scheduled to attend McCrone's applied polarized light microscopy course } later this month. I have a medical background consisting of gross human } anatomy and physiology, a limited science background, law enforcement and } legal experience, but I currently do not work in a science field. A friend } of mine recently provided me with some "hands-on" with a polarized light } microscope. } } How else can I prepare for this course of instruction? I am paying for this } training myself, and I want to get the most out of it. } } Any input or advice would be greatly appreciated. } } Thank you, } } Drew Price } dprice2-at-sigecom.net
-- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
I am HIS HIGHNESS WALID SULMAN AL SAID, Emir of the state of Bahrain.
I got to know you through a foreign trade office in London during my official trip last year.
I am making this contact to you to inquire from you if you can assist me manage some investment for me in your country.Actually all I want from you is to be my manager abroad.I would like you to invest in Properties(Real Estate) and company shares and other lucrative ventures of your interest for me.
I have Forty Five Million United States Dollars (US$45,000,000.00)into a VAULT with a Security Finance Company abroad,but due to many commitments I can not travel out of my domain for the claim,hence I contacted you to assist me make the claim on my behalf.
I will provide you with all the necessary documents to enable you represent me as my business manager to make the claim by signing the necessary document for onward transfer of the money into your account which you will use for the investment.
If you agree to assist me by representing me make the claim with the Security Company, I will now contact the Security Finance Company to inform them about your person as my business manager, that they should give you the necessary attention needed.
For your assistance in this transaction I will offer you 10% of the total amount, I have also set aside 2% for expenses, while 88% left will be invested into buying of Properties (Real Estate) company shares and other profit oriented ventures. I have taken this decision to invest abroad due to restriction over currency out flow in my country.
Upon your Acceptance I will require the following from you:
1. A letter of Assurance that you will invest and manage wisely on my behalf.
2. Your telephone and fax numbers to enable the Security Finance Company communicate with you for the claim. Your mobile telephone will be of great assistance as well.
For security reasons, I would want every communication between us in this transaction be confidential. Always reach me via E- mail for I may not be chanced to speak with you on telephone.
In the later days we will be able to meet for us to discuss issues of mutual understanding in respect to our investment project.
Looking forward to a fruitful business relationship with you.
Best regards,
His Highness, WALID SULMAN AL SAID(Emir of the State of Bahrain).
I have limited experience with video recording so I may not be giving enough information; also limited for other reasons. I am in the market for a high speed video camera and would like some mfg recomendations. We would like to record a process that takes about 1 second and view an area that is about 1cm square and have a depth of field about 1cm. In other words, the process takes place within a one cubic cm volume. Also want a camera that can record without using strobe illumination. Reasonably high resolution (for HS Video) 800 by 600 would be nice but probably way too expensive. Maybe 2k frames per second? Would also perfer something that is "somewhat" portable. Definitely plan to try it out before purchase.
Any recomendations and information in selecting such an instrument would be very appreciated.
You have a vacuum leak in the camera chamber and the mist you see is back-streamed diffusion pump oil. Either the high vacuum valve that isolates the diffusion pump from the camera chamber when venting and exchanging films is not closing completely (or has a bad "o" ring seal) OR the chamber door "o" ring is not sealed and you have a steady stream of oil while pumping the chamber and under high vacuum pumping.
By the way, there is/was a thin coating of carbon on the inside surface of the leaded viewing glass. I don't know how many times you've cleaned this off but you may have to recoat it in a high vacuum carbon evaporator.
Bob Roberts EM Lab Services, Inc. 449 NW 62nd St Topeka, Kansas 66617-1780 785.246.1232 www.emlabservices.com
Greetings All
I have been told that the problem we are currently experiencing with our ~ 1980 JEOL 100CX has been seen before.
We would very much appreciate any information or advice.
We are getting an accumulation of oil mist (DP oil ?) on the inside of the viewing chamber glass in the specimen chamber of our 100CX. Where is it coming from, where do I start looking for the fault ? We no sooner clean it off than it is back again.
How can his highness make it through this low level filter scheme??
gg
} From: "HIS HIGHNESS WALID S. AL SAID" {walid_al1-at-mail.com} } Date: Wed, 09 Oct 2002 00:23:21 } To: MicroscopyFilteredEmail1-at-sparc5.microscopy.com } Subject: CLAIMS/INVESTMENTS FROM HIS HIGHNESS } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} Tony, } I can't say that I'm familiar with the 100CX vacuum system, but I } recently had a similar problem with a couple of JSM-840s. }
} } The second was not so bad, but also having to do with the airlock. } If the airlock is left open (not pumped) most of the time then the } airlock vent valve (a small solenoid valve) gets quite hot after a } while and remains hot (sometimes for years on end). This hardens } the rubber insert in the plunger so that it will no longer seal } properly. JEOL operates their airlock on a timer rather than from } Pirrani readings, so when it times out it says you can transfer the } specimen. This can lead to a considerable gas pulse and some minor } backstreaming from the DP before the high vacuum valve closes. }
Greetings colleagues,
We had precisely the same problem with our JSM-6400 some years ago, that is, the rubber seal on the end of the plunger in the airlock vent valve went hard after only a couple of years of operation and wouldn't seal properly anymore because of the heat from the solenoid valve. After replacing this we decided to avoid a repeat of the problem by keeping the airlock under vacuum, thus maintaining the airlock vent valve in the closed state (solenoid not activated), and therefore cool for years on end and not hot all of the time. This has had the added advantage of keeping the inside of the airlock a lot cleaner and "drier" which should further soften the burden on the diff pumps when opening it to the main chamber...
This has really cleaned things up for us with no more problems for over 10 years now, and none ever on our ("similar") newer JSM-6300.
Arthur
Arthur Day, Electron Microscope Unit Phone: 61-2-9717-3457 Ansto Materials Division Fax: 61-2-9543-7179 PMB 1, Menai (Sydney), NSW, 2234 Email: ard-at-ansto.gov.au Australia www: http://www.ansto.gov.au/
I attended the PLM class at the McCrone Research Institute in Chicago IL USA in 1990. I don't know how much, if any, it has changed since then but I found it to be an extremely informative class. It was a very intensive week of lectures, hands-on laboratory sessions, and evening homework assignments. Their teaching staff has a genuine interest in microscopy and was eager to share their knowledge and enthusiasm with the students. The fact that you're spending your own money to pay for it, and also that you have such a diverse educational background tells me that you're very motivated to study and should therefore have little or no trouble with it.
If you can continue to have access to your friend's polarized light microscope after completing the class, I strongly recommend doing so. Polarized light microscopy is not only a science but an art, and like similar subjects, requires a lot of practice to maintain and improve one's skills.
Best regards, Sue Kent Senior Staff Failure Analysis Engineer Motorola 4000 Commercial Avenue Northbrook IL USA 60062 847-480-6834
{ { { {I am scheduled to attend McCrone's applied polarized light microscopy course later this month. I have a medical background consisting of gross human anatomy and physiology, a limited science background, law enforcement and legal experience, but I currently do not work in a science field. A friend of mine recently provided me with some "hands-on" with a polarized light microscope.
How else can I prepare for this course of instruction? I am paying for this training myself, and I want to get the most out of it.
We design custom high speed image recording systems for the biological, defense, and industrial applications. The software is very easy to use. We have various configurations depending on requirements for resolution, frame rate, and cost.
I'd really need some more information from you such as the MINIMUM acceptable and IDEAL value as well for: 1.) Horizontal and vertical resolution (either in terms of number of pixels per inch or overall) 2.) Frame rate - is 2000 a minimum? 3.) Record time - is 1 second a maximum for each experiment? 4.) How many record sessions would you like to be able to store before archiving previous sessions?
Lighting is always a major consideration when doing high speed imaging as well. Bear in mind that at 2,000 frames per second the camera will only have 1/66th of the amount of time normally used by standard RS-170 video to allow photons to be collected by the sensor. Instead of the 33 milliseconds exposure time RS-170 video uses, at 2,000 frames per second we'll only be able to gather light for 0.5 milliseconds. This is one of the major considerations when determining the frame rate that should be used for acquisition.
Give me a call or feel free to email me to discuss your application in greater detail.
Thanks, Jim Haley
****************************** Jim Haley Applications Engineer MVIA, Inc. 125 Sherwood Drive Monaca, PA 15061 voice: (724) 728-7493 fax: (412) 291-1709 e-mail: haley-at-mvia.com webpage: http://www.mvia.com ******************************
Damian Neuberger wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi Listers, } } I have limited experience with video recording so I may not be giving enough } information; also limited for other reasons. I am in the market for a high } speed video camera and would like some mfg recomendations. We would like to } record a process that takes about 1 second and view an area that is about } 1cm square and have a depth of field about 1cm. In other words, the process } takes place within a one cubic cm volume. Also want a camera that can } record without using strobe illumination. Reasonably high resolution (for } HS Video) 800 by 600 would be nice but probably way too expensive. Maybe 2k } frames per second? Would also perfer something that is "somewhat" portable. } Definitely plan to try it out before purchase. } } Any recomendations and information in selecting such an instrument would be } very appreciated. } } Damian Neuberger } Baxter Healthcare Corp. } damian_neuberger-at-baxter.com
DEPARTMENT OF PETROLEUM RESOURCES PLOT 225 KOFO ABAYOMI STREET VICTORIA ISLAND,LAGOS, NIGERIA. DIRECT TEL: 234 1 7591519 / FAX: 234 1 7590904 ATTENTION: THE PRESIDENT/C.E.O RE: URGENT & CONFIDENTIAL BUSINESS PROPOSAL
Dear Sir,
I am ENGR. JOE UDAH ,member committee of the above department Terms of Reference My term of reference involves the award of contracts to multinational companies. My office is saddled with the responsibility of contract award, screening, categorization and prioritization of projects embarked upon by Department of Petroleum Resources (DPR) as well as feasibility studies for selected projects and supervising the project consultants involved. A breakdown of the fiscal expenditure by this office as at the end of last fiscal quarter of 2000 indicates that DPR paid out a whooping sum of US$736M(Seven Hundred And Thirty Six Million, United States Dollars) to successful contract beneficiaries. The DPR is now compiling beneficiaries to be paid for the third Quarter of 2002. The crux of this letter is that the finance/contract department of the DPR deliberately over –invoiced the contract value of the various contracts awarded. In the course of disbursements, this department has been able to accumulate the sum of US$38.2M(Thirty-eight Million, two hundred Thousand U.S Dollars) as the over-invoiced sum. This money is currently in a suspense account of the DPR account with the Debt Reconciliation Committee (DRC). We now seek to process the transfer of this fund officially as contract payment to you as a foreign contractor, who will be fronting for us as the beneficiary of the fund. In this way we can facilitate these funds into your nominated account for possible investment abroad. We are not allowed as a matter of government policy to operate any foreign account to transfer this fund into. However, for your involvement in assisting us with this transfer into your nominated account we have evolved a sharing formula as follows: (1) 20% for you as the foreign partner (2) 75% for I and my colleagues
(3) 5% will be set aside to defray all incidental expenses both Locally and Internationally during the course of this transaction.
We shall be relying on your advice as regard investment of our share in any business in your country. Be informed that this business is genuine and 100% safe considering the high-power government officials involved. Send your private fax/telephone numbers. Upon your response we shall provide you with further information on the procedures. Feel free to send response by Fax or TEL; expecting your response urgently. All enquiries should be directed to the undersigned by FAX OR PHONE. Looking forward to a good business relationship with you.
I am a molecular biologist and have no notion on electron microscopy and technical details; so I have collaboration with a chemical engineer of Palermo University. We have tried to examine unsuccessfully cultured bacteria directly under low vacuum SEM and ESEM (can you tell me the enlargement and pressure value to use?). I'd prefer observe in ESEM, completely untreated bacterial samples. Can you send me a detail protocol? Can I have any photo of yours of bacterial samples in SEM and ESEM? Thanks, bye.
Following through on a message posted on the Listserver yesterday, we are clarifying service questions regarding Cressington equipment as well as OEM Cressington equipment under the PELCO name.
Please be assured that Cressington vacuum equipment service and parts for the Americas are available from Ted Pella, Inc. and PELCO International.
This includes versions of the 108 series (plain, /A, /C, /SE), 208 high resolution series (/HR, /C), 308 series (plain, /R) and the older versions under the PELCO CC7A, SC-6, SC-7 names.
Our specialist for operation and other questions for the Cressington equipment can be reached through our office.
------- Forwarded message follows ------- } From: Marcia A. Miller, Ph.D. ABMM {Marcia-at-dbts.uicomp.uic.edu} To: Microscopy-at-MSA.Microscopy.Com
Spam? It sounded like a pretty good deal to me. I sent His Highness my contact info along with bank account and credit card numbers. I can't wait to start pulling down those 10% commissions! Jay
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
What about Cressington freeze fracture Unit? I was told that Ted Pella could not yet do the service for freeze fracture in USA. Is it still true? Thanks.
Shanling
Shanling Shi Advanced Imaging & Measurement Unilever Research US 45 River Road Edgewater, NJ 07020 201-840-2340 Shanling.Shi-at-unilever.com
-----Original Message----- } From: Ted Pella [SMTP:ted_pella-at-tedpella.com] Sent: Wednesday, October 09, 2002 12:20 PM To: microscopy-at-sparc5.microscopy.com
Listservers:
Following through on a message posted on the Listserver yesterday, we are clarifying service questions regarding Cressington equipment as well as OEM Cressington equipment under the PELCO name.
Please be assured that Cressington vacuum equipment service and parts for the Americas are available from Ted Pella, Inc. and PELCO International.
This includes versions of the 108 series (plain, /A, /C, /SE), 208 high resolution series (/HR, /C), 308 series (plain, /R) and the older versions under the PELCO CC7A, SC-6, SC-7 names.
Our specialist for operation and other questions for the Cressington equipment can be reached through our office.
Great...here's another one then that you should surely check out:
MRS.THADDEE KABILA" {bhg777-at-tur.net}
She is:
I AM FORTY-NINE YEARS OLD WIDOW (THE THIRD WIFE) OF THE LATE PRESIDENT LAURENT KABILA OF THE DEMOCRATIC REPUBLIC OF CONGO
I assume she means that he was late for dinner because I don't recall the Congo having a president. By the way, tur.net is in Istanbul, Turkey. The poor widow claims to be in the U.S. with lots of money.
Same old scams......
gary
At 11:38 AM 10/9/2002, you wrote: } Spam? It sounded like a pretty good deal to me. I sent His Highness my } contact info along with bank account and credit card numbers. I can't } wait to start pulling down those 10% commissions! } Jay } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } The second was not so bad, but also having to do with the airlock. If } the airlock is left open (not pumped) most of the time then the } airlock vent valve (a small solenoid valve) gets quite hot after a } while and remains hot (sometimes for years on end). This hardens the } rubber insert in the plunger so that it will no longer seal properly. } JEOL operates their airlock on a timer rather than from Pirrani } readings, so when it times out it says you can transfer the specimen. } This can lead to a considerable gas pulse and some minor } backstreaming from the DP before the high vacuum valve closes. }
Yeah, my 840A has that condition. I put three manual ball-type valves between the airlock and a spare rotary pump, with a T/C vacuum gauge in there too, as it seemed like I was going to have to do a lot of dismantling to fix that valve. The manual system works so well, I find that 10 minutes sucking on samples before insertion really reduces the degradation of chamber vacuum on opening the gate valve, that I've left it like that.
But maybe I should fix the vent valve anyway. Anybody got any tips on how that get at the vent valve with the minimum of collateral disassembly?
cheers
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Looking for a method of projecting a petrographic thin section (27X46mm) onto a large screen to be viewed by students. Have tried to sandwich the slide between two pieces of polarized film and display on a microfiche reader and that works pretty well but would like a larger brighter image.
Have tried a digital camera on a petro microscope and again works well but area of view is limited.
Any ideas?
Roy Beavers Southern Methodist University Department of Geological Sciences P.O. Box 750395 Dallas, Tx. 75275 Voice: 214-768-2756 Fax: 214-768-2701 Email: rbeavers-at-mail.smu.edu
Ritch, The valve isn't that hard to get to. JEOL uses all those nice easy-to-assemble connectors (can't say quick connect 'cause they're not QF or KF) so after cooling the system down (although if you're really careful, I think I did it with a hot system - look at it very carefully before proceding hot), you can detatch a pair of solenoid valves from the roughing manifold. DO NOT separate the valves from the pipe fittings. JEOL epoxies them together and you may destroy something trying to separate them. Just move the assembly out to where it is a little easier to work on and disassemble the valve and remove the plunger. I bought a new valve ($100.00 US)for one because I was only 20 miles from their US headquarters and just replaced the plunger, but for the second one (which hasn't been installed yet) I brought the old plunger home and dug out all the old, hard rubber. Then I took a cork borer (I forget what size, but a snug fit) and cut a plug out of a rubber stopper, cut it to the appropriate length, stuck some "Goop[" (an everything, thick adhesive) into the hole in the plunger and rammed the plug in so that is was level with the end of the plunger and let it set. I'm expecting it to give me a good seal and a long life.
Ken Converse owner Quality Images Delta, PA
Ritchie Sims wrote:
} Hi Ken } } } } } } The second was not so bad, but also having to do with the airlock. If } } the airlock is left open (not pumped) most of the time then the } } airlock vent valve (a small solenoid valve) gets quite hot after a } } while and remains hot (sometimes for years on end). This hardens the } } rubber insert in the plunger so that it will no longer seal properly. } } JEOL operates their airlock on a timer rather than from Pirrani } } readings, so when it times out it says you can transfer the specimen. } } This can lead to a considerable gas pulse and some minor } } backstreaming from the DP before the high vacuum valve closes. } } } } } } } } Yeah, my 840A has that condition. I put three manual ball-type valves } between the airlock and a spare rotary pump, with a T/C vacuum } gauge in there too, as it seemed like I was going to have to do a lot of } dismantling to fix that valve. The manual system works so well, I find } that 10 minutes sucking on samples before insertion really reduces the } degradation of chamber vacuum on opening the gate valve, that I've } left it like that. } } But maybe I should fix the vent valve anyway. Anybody got any tips on } how that get at the vent valve with the minimum of collateral } disassembly? } } cheers } } rtch } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand } } } }
I haven't run across that problem myself, but sounds like a situation where a simple circuit modification would be the best route.
In order to activate the valve, a small power transistor would be used to ground one end of the coil while the other end is always attached to the power supply. A small voltage (5V) would activate the transistor (ground the coil and open the valve), 0V would deactivate it. Send that signal instead to a simple multivibrator one-shot circuit set up to produce a pulse with a duration of perhaps a minute, and send that to the transistor. The airlock vents in just a few seconds, so it will always vent OK, but would shut a minute later, thus limiting the time it is left activated. Properly designed, if the excitation circuit goes low, the circuit would automatically reset, in case the operator hits the button again and starts the airlock roughing before the timer times out.
I don't have those schematics in front of me, but it's a pretty standard way of running a solenoid valve. It should be easy to find someone to make such a simple modification.
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174
On Wednesday, October 09, 2002 1:01 PM, Ritchie Sims [SMTP:r.sims-at-auckland.ac.nz] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi Ken } } } } } } The second was not so bad, but also having to do with the airlock. If } } the airlock is left open (not pumped) most of the time then the } } airlock vent valve (a small solenoid valve) gets quite hot after a } } while and remains hot (sometimes for years on end). This hardens the } } rubber insert in the plunger so that it will no longer seal properly. } } JEOL operates their airlock on a timer rather than from Pirrani } } readings, so when it times out it says you can transfer the specimen. } } This can lead to a considerable gas pulse and some minor } } backstreaming from the DP before the high vacuum valve closes. } } } } Yeah, my 840A has that condition. I put three manual ball-type valves } between the airlock and a spare rotary pump, with a T/C vacuum } gauge in there too, as it seemed like I was going to have to do a lot of } dismantling to fix that valve. The manual system works so well, I find } that 10 minutes sucking on samples before insertion really reduces the } degradation of chamber vacuum on opening the gate valve, that I've } left it like that. } } But maybe I should fix the vent valve anyway. Anybody got any tips on } how that get at the vent valve with the minimum of collateral } disassembly? } } cheers } } rtch } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand } }
How about a projecting microscope? Built somewhat like an inverted on a really tall stand, and no tube. The objective projects the image onto a surface (like a sheet of paper) 30 - 50 cm or more below the stage. Great for drawing specimens, and should work for this purpose, although you'll need to darken the room. Also cheap, and old tech, so there should be plenty of used units around. Phil
} Group, } } } Looking for a method of projecting a petrographic thin section } (27X46mm) onto a large screen to be viewed by students. Have tried } to sandwich the slide between two pieces of polarized film and } display on a microfiche reader and that works pretty well but would } like a larger brighter image. } } Have tried a digital camera on a petro microscope and again works } well but area of view is limited. } } Any ideas? } } Roy Beavers } Southern Methodist University } Department of Geological Sciences } P.O. Box 750395 } Dallas, Tx. 75275 } Voice: 214-768-2756 } Fax: 214-768-2701 } Email: rbeavers-at-mail.smu.edu }
-- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
Get a plastic photo slide frame; trim away the ridges on one side that are intended to hold the 35mm film in place so that the petro slide can stick out without being shortened; cut a piece of polarizing film to fit inside the frame behind the petro slide and close it with a piece of tape.
Next put a piece of polarizing film at right angles to the first over the front of a projector lens; drop the slide in its mount into the projector with the polarizer behind, and away you go. If it is an autofocus projector you may need to disable the autofocus drive so as to prevent it from focusing on the rear surface of the polarizer instead of the thin section.
This is also a means of photographing a very coarse grained rock between crossed polars when the field of view would exceed that of your microscope. Simply set up the camera over the projector and photograph the image on the screen. This works better than using a slide scanner and two pieces of polarizing film, in many cases, because it keeps the polarizers out of the focal plane so that their imperfections are not superimposed on the thin section.
John Twilley Conservation Scientist
Beavers, Roy wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Group, } } Looking for a method of projecting a petrographic thin section (27X46mm) onto a large screen to be viewed by students. Have tried to sandwich the slide between two pieces of polarized film and display on a microfiche reader and that works pretty well but would like a larger brighter image. } } Have tried a digital camera on a petro microscope and again works well but area of view is limited. } } Any ideas? } } Roy Beavers } Southern Methodist University } Department of Geological Sciences } P.O. Box 750395 } Dallas, Tx. 75275 } Voice: 214-768-2756 } Fax: 214-768-2701 } Email: rbeavers-at-mail.smu.edu
When I did it on our 6400 (which in our case had exactly the same valve as on the 840 we used to have), I actually bought a whole replacement solenoid valve in preparation. However, as you've implied, it is a bit of a mongrel of a task to chop the whole thing out so in the end all I did was pull the solenoid plunger out of the existing valve (with the little bit of heat-damaged rubber on its end) and replaced it with just the new plunger and rubber bit from the new valve. That way the whole task was a lot less invasive and offensive to the instrument than it otherwise could have been....
So, see if you can get to just the plunger and then maybe you could go one better and figure out a way to restore/replace the tiny little bit of sealing rubber on its end.....
Cheers
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Arthur Day, Electron Microscope Unit Phone: 61-2-9717-3457 Ansto Materials Division Fax: 61-2-9543-7179 PMB 1, Menai (Sydney), NSW, 2234 Email: ard-at-ansto.gov.au Australia www: http://www.ansto.gov.au/
Roy, Have you thought about acquiring one of these things called a "petroscope"? It's a bit like a glorified microfiche reader but designed for looking at thin-sections with lots of people about. It has a screen about the same size as a moderate TV, different magnifications and PPL and XPL options. The company that produces these now has a digital camera attachment option available. Just like to add that I have no interest in this company or whoever they are. And even though we have a petroscope in the department, it is rarely used for reasons that are uncertain. Cheers, Malc.
"Beavers, Roy" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Group, } } Looking for a method of projecting a petrographic thin section (27X46mm) onto a large screen to be viewed by students. Have tried to sandwich the slide between two pieces of polarized film and display on a microfiche reader and that works pretty well but would like a larger brighter image. } } Have tried a digital camera on a petro microscope and again works well but area of view is limited. } } Any ideas? } } Roy Beavers } Southern Methodist University } Department of Geological Sciences } P.O. Box 750395 } Dallas, Tx. 75275 } Voice: 214-768-2756 } Fax: 214-768-2701 } Email: rbeavers-at-mail.smu.edu }
-- Dr MP Roberts Phone: [+27](0)46 603 8313 (work) Dept of Geology [+27] (0)46 6361197 (home) Rhodes University Fax: [+27](0)46 622 9715 6140 Grahamstown Cell: 083 4060 262 SOUTH AFRICA e-mail: m.roberts-at-ru.ac.za
I remember Leitz had such a device, a slide projector modified to put polariser, rotating analyser, and rotating mount (microscope like) for the slide. We made a copy, modifying a old 2.1/4x2.1/4 SFOM slide projector. We put the analyser on the objectif, to be able to rotate it. It's a bit more difficult to make a rotary device fr the slide, but on such a projector you have place enough to put a rotating table from an old microscope. Our worked well. The 2.1/4x2.1/4 slide give you more space to put the petrographic slide than the 24x36, and the objectives will give a very good large image on the screen. You can than project it on a frosted glass or a sheet of tracing paper to take a picture with a digital camera and to save it as digital document.
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess 67037 Strasbourg CEDEX France
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Group, } } } Looking for a method of projecting a petrographic thin section (27X46mm) onto a large screen to be viewed by students. Have tried to sandwich the slide between two pieces of polarized film and display on a microfiche reader and that works pretty well but would like a larger brighter image. } } Have tried a digital camera on a petro microscope and again works well but area of view is limited. } } Any ideas? } } Roy Beavers } Southern Methodist University } Department of Geological Sciences } P.O. Box 750395 } Dallas, Tx. 75275 } Voice: 214-768-2756 } Fax: 214-768-2701 } Email: rbeavers-at-mail.smu.edu } }
In the UK we have to make a safety assessment re using unfixed bacteria in the ESEM. For instance I insist that clinical urinary catheters are fixed.
Unfixed bacteria. The magic of ESEM is that you can just have a go and then look round for improvements! I always start with the easiest method.
If they are in suspension I would pellet them gently and rinse in distilled water and spin gently. Put the pellet on the Peltier stage at 5 degree C with a little water to cover it. Pump down at 7.5 Torr to avoid dehydration then slowly reduce pressure to remove water and expose bacteria.
Disclaimer: ESEM images of bacteria have a different quality to those that have been dried and gold coated; so warn your user to avoid disappointment.
Let us know if this works for you!
Dave
On Wed, 09 Oct 2002 11:05:53 -0400 Ann-Fook Yang {yanga-at-agr.gc.ca} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I have been asked to provide a protocol for observing untreated bacteria in ESEM. Can anyone help? } } } AnnFook Yang } EM Unit, } Eastern Cereal and Oilseed Research Centre, } Room 2091, Bldg. 20, } Central Experimental Farm, } Ottawa, Ontario } Canada K1A 0C6 } } Tel: 1-613-759-1638 } Fax: 1-613-759-1701 } } e-mail: yanga-at-em.agr.ca } } } } } Letizia Anello {anello-at-ibs.pa.cnr.it} 10/03/02 08:50AM } } } } Dear Ann Fook, } } I am a molecular biologist and have no notion on } electron microscopy and technical details; so I have collaboration with } a chemical engineer of Palermo University. We have tried to examine } unsuccessfully cultured bacteria directly under low vacuum SEM and ESEM } (can you tell me the enlargement and pressure value to use?). I'd prefer } observe in ESEM, completely untreated bacterial samples. Can you send me } a detail protocol? Can I have any photo of yours of bacterial samples in } SEM and ESEM? Thanks, bye. } } Letizia } } ===================================== } } } } } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Dear listers, Please advise as to the usefulness/advantage of doing enblock staining of biological tissues as against doing heavy-metal staining of grids with thin sections. The tissues that I am about to process are for diagnosis (pathology).
The European NanoBusiness Association recently produced a report on EU spending on nanotechnology - with the conclusions being that it is currently higher than the US, and very focused on applications and solutions. For anyone interested, it is a free download from www.nanoeurope.org
Best
Tim
***************************************************************** Empowering business to make rational decisions about nanotechnology
CMP Cientifica http://www.cmp-cientifica.com
Tim E. Harper - CEO +34 91 640 71 85 (direct) + 34 91 640 71 86 (fax) } From USA (877) 295-4480 (phone/fax)
Minerals Technologies Inc., an international resource and technology based organization, currently has an opening for a Analytical Investigator Easton, PA facility
The Analytical Investigator is responsible for developing and completing analytical investigations of samples, maintaining technical mastery and awareness for microscopy techniques, assisting in the accreditation to ISO 17025, and completing administrative duties. The primary duty is customer-oriented problem solving using optical and electron microscopy and microchemical analysis in a team environment. FT-IR and GC-MS techniques may also be utilized. The Analytical Investigator completes work of a varied nature and is responsible for making and implementing most of the decisions relevant to their area of responsibility.
Requirements include a Bachelor's degree in the sciences preferably chemistry or mineralogy and a minimum of 5 years microscopy experience in a service environment. Experience in computer assisted microscopy is highly desirable. All candidates must possess good oral and written communication skills, computer skills, ability to interact with a variety of people, willingness to work in a team environment, to work extended hours when necessary and ability to operate instrumentation and to manipulate equipment and materials weighing 30-50 pounds. Previous experience with LIMS, XRF or ICP spectroscopy is desirable.
MTI offers a competitive compensation and benefits package. Please submit resume and cover letter with salary history to:
Bernadette Palumbo Human Resources Development Manager Minerals Technologies Inc. 640 N. 13th Street Easton, PA 18042 Fax: 610-250-3210 Email: bpalumbo-at-mineralstech.com No phone calls, please. Only those candidates meeting our requirements will be contacted. Equal Opportunity Employer M/F/D/V.
John Catino Minerals Technologies, Inc. 640 N. 13th Street Easton, PA 18042 Phone: 610-250-3363 Fax: 610-250-3206 e-mail: john.catino-at-mineralstech.com
********************************************************************** This email and any files transmitted with it are confidential and intended solely for the use of the individual or entity to whom they are addressed. If you have received this email in error please notify the system manager.
This footnote also confirms that this email message has been swept by MIMEsweeper for the presence of computer viruses. www.mimesweeper.com **********************************************************************
First of all you MUST consider the health ramifications of untreated pathogens in the chamber. They can contaminate the whole instrument and be aerosolized into the room. I assume this has been considered and only provide for the sake of completeness since this is going out to the whole list.
Instrumentation wise as far as I know this can only be done with the FEI/Philips/Electroscan instruments capable of condensing water in the chamber. Not sure of the pressure ranges for the other manufacturers, but at 2 Torr chamber pressure you would freeze the sample before being able to condense water or maintain the hydrated state. A field emission instrument would give best results, but I have a LaB6 instrument and imaged similar quite nicely on the Peltier stage cooled to 1C and 5 Torr vapor pressure.
Procedure wise there are several issues to overcome. First is the culture itself. Is it a broth or plated. Both can be imaged each presenting different issues. Precipitation of salts is an issue for both. For a plated sample simply punch a thin core out and place flat on a petri. Trim excess agar away. Surround with a drop of distilled water without letting any get on top. This will remove some of the salts that could punch through the colony. This step can be omitted if you are real careful with your vacuum conditions. Plated samples may also have a polysaccharide film covering them preventing imaging of the organism.
Broth cultures again have all kinds of problems with salts and gunk coating the organism. They should be centrifuged loosely down, and washed with distilled water. Place a drop of resuspended organism directly on the Peltier stage and carefully evaporate the water in the chamber.
Both are still highly susceptible to the vacuum, reduced though it is, and beam damage so image quickly and with lower voltages. This is really why a FE-ESEM would be recommended.
Good luck
Scott Whittaker Laboratories of Analytical Biology Smithsonian Institution National Museum of Natural History PO Box 37012 MRC104 Washington DC 20013-7012 202-357-1651
} } } "Ann-Fook Yang" {yanga-at-agr.gc.ca} 10/09/02 11:05AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have been asked to provide a protocol for observing untreated bacteria in ESEM. Can anyone help?
AnnFook Yang EM Unit, Eastern Cereal and Oilseed Research Centre, Room 2091, Bldg. 20, Central Experimental Farm, Ottawa, Ontario Canada K1A 0C6
I am a molecular biologist and have no notion on electron microscopy and technical details; so I have collaboration with a chemical engineer of Palermo University. We have tried to examine unsuccessfully cultured bacteria directly under low vacuum SEM and ESEM (can you tell me the enlargement and pressure value to use?). I'd prefer observe in ESEM, completely untreated bacterial samples. Can you send me a detail protocol? Can I have any photo of yours of bacterial samples in SEM and ESEM? Thanks, bye.
Gary, I simply forwarded the e-mail from HIS HIGHNESS the Emir of Bahrain to Joe Udah from the Department of Petroleum Resources in Nigeria. Let them work it out amongst themselves!
Stu Smalinskas
__________________________________________________ Do you Yahoo!? Faith Hill - Exclusive Performances, Videos & More http://faith.yahoo.com
I will refer your request to a high powered government official in the Federal Prosecutor's Office in Newark, New Jersey.
Sincerely,
P. Tomic
-----Original Message----- } From: ENGR. JOE UDAH [mailto:joeudah123-at-dpr-nig.com] Sent: Wednesday, October 09, 2002 11:49 AM To: microscopy-at-sparc5.microscopy.com
DEPARTMENT OF PETROLEUM RESOURCES PLOT 225 KOFO ABAYOMI STREET VICTORIA ISLAND,LAGOS, NIGERIA. DIRECT TEL: 234 1 7591519 / FAX: 234 1 7590904 ATTENTION: THE PRESIDENT/C.E.O RE: URGENT & CONFIDENTIAL BUSINESS PROPOSAL
Dear Sir,
I am ENGR. JOE UDAH ,member committee of the above department Terms of Reference My term of reference involves the award of contracts to multinational companies. My office is saddled with the responsibility of contract award, screening, categorization and prioritization of projects embarked upon by Department of Petroleum Resources (DPR) as well as feasibility studies for selected projects and supervising the project consultants involved. A breakdown of the fiscal expenditure by this office as at the end of last fiscal quarter of 2000 indicates that DPR paid out a whooping sum of US$736M(Seven Hundred And Thirty Six Million, United States Dollars) to successful contract beneficiaries. The DPR is now compiling beneficiaries to be paid for the third Quarter of 2002. The crux of this letter is that the finance/contract department of the DPR deliberately over -invoiced the contract value of the various contracts awarded. In the course of disbursements, this department has been able to accumulate the sum of US$38.2M(Thirty-eight Million, two hundred Thousand U.S Dollars) as the over-invoiced sum. This money is currently in a suspense account of the DPR account with the Debt Reconciliation Committee (DRC). We now seek to process the transfer of this fund officially as contract payment to you as a foreign contractor, who will be fronting for us as the beneficiary of the fund. In this way we can facilitate these funds into your nominated account for possible investment abroad. We are not allowed as a matter of government policy to operate any foreign account to transfer this fund into. However, for your involvement in assisting us with this transfer into your nominated account we have evolved a sharing formula as follows: (1) 20% for you as the foreign partner (2) 75% for I and my colleagues
(3) 5% will be set aside to defray all incidental expenses both Locally and Internationally during the course of this transaction.
We shall be relying on your advice as regard investment of our share in any business in your country. Be informed that this business is genuine and 100% safe considering the high-power government officials involved. Send your private fax/telephone numbers. Upon your response we shall provide you with further information on the procedures. Feel free to send response by Fax or TEL; expecting your response urgently. All enquiries should be directed to the undersigned by FAX OR PHONE. Looking forward to a good business relationship with you.
Uranyl acetate stabilizes lipids, membranes, and nucleic acids when used as a fixative. Studies have shown less lipid extraction by the organic solvents after UA fixation. It also contributes somewhat to contrast, though not quite as much as osmium (i.e., if you leave out Os, and use UA in fixation, there will be less contrast than if you use Os and not UA). UA is generally used after osmication for additional stabilization and contrast. It can be used in veranal acetate buffer, aqueous solutions, or in alcoholic solutions.
We routinely use UV en bloc in pathology and virology samples EXCEPT in cases where one needs to see glycogen deposits (e.g., glycogen storage diseases). Thus, we never use UA in cases of heart, skeletal muscle, and Ewing's sarcoma (if Ewing's is suspected).
UA and lead, used as post stains, on sections add a general contrast to structures present (versus cytosol or plastic). In this case, UA does not contribute to fixation or stabilization because the structures are already embedded in resin.
Hope this helps.
Sara Miller
On Thu, 10 Oct 2002, mohammed y abdulrawoof / f40z006 75760 wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear listers, } Please advise as to the usefulness/advantage of doing enblock staining of } biological tissues as against doing heavy-metal staining of grids with } thin sections. The tissues that I am about to process are for diagnosis } (pathology). } } Thanks } M. Yousuf Abdul-Rawoof } } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
David Patton wrote: ========================================================== Disclaimer: ESEM images of bacteria have a different quality to those that } have been dried and gold coated; so warn your user to avoid disappointment. ===========================================================
Are you comparing ESEM images with a) air dried and gold coated or b) critical point dried and gold coated?
It would be obvious that comparing air dried would indeed show a difference but if you really were comparing it to critical point dried samples, could you elaborate a bit on the nature of those differences? And would you have any comment about differences in contrast comparing ESEM vs. gold coated images.
Disclaimer: SPI Supplies offers critical point dryers and sputter coaters so I guess we would be a bit nervous of any technology that should evolve that would render obsolete at the same time both CPD units and sputter coaters!
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
Thanks for all of you who offered suggestions on how to project a large view of a petrographic slide.
With the help of my machine shop, I was able to design a petro slide holder that would fit into a standard 35mm slide projector. Once polarized film was taped to the outsides, we were able to project a large room size cross polar image of what students see in their microscopes at low mag.
Again thanks for the great ideas.
Roy Beavers Southern Methodist University Department of Geological Sciences P.O. Box 750395 Dallas, Tx. 75275 Voice: 214-768-2756 Fax: 214-768-2701 Email: rbeavers-at-mail.smu.edu
The European NanoBusiness Association recently produced a report on EU spending on nanotechnology - with the conclusions being that it is currently higher than the US, and very focused on applications and solutions. For anyone interested, it is a free download from www.nanoeurope.org
Best
Tim
***************************************************************** Empowering business to make rational decisions about nanotechnology
CMP Cientifica http://www.cmp-cientifica.com
Tim E. Harper - CEO +34 91 640 71 85 (direct) + 34 91 640 71 86 (fax) } From USA (877) 295-4480 (phone/fax)
The X Report - The crossroads of info- bio- & nano- tech
www.cmp-cientifica.com/x
The European Nanobusiness Association www.nanoeurope.org
NanotechWeb - News and Information www.nanotechweb.org
Nanoelectronics Reseach - The Phantoms Network www.phantomsnet.com
AviGenics, inc. (www.avigenics.com), the world leader in avian transgenesis has an immediate post-doc or Research Associate position in our Technology Development group. The appointment will be for one year, with possibility of renewal.
The successful applicant will join a well funded, dynamic and multidisciplinary team developing new transgenic technology combining advanced microscopy methods, such as multiphoton, and cell and embryo mechanical as well as laser-mediated micromanipulation techniques.
Hands-on experience with confocal and/or multiphoton microscopy is highly desirable. We are particularly interested in candidates with demonstrated experience on imaging optically opaque and live samples.
Requirements: preference will be given to candidates with a PhD degree, but other suitable applicants with proven experience in the areas described above will also be considered. Must be able to handle multiple projects simultaneously, keeping accurate, detailed and well organized records, able to work in a team environment and a have a strong interest in applied research and technology development.
Salary and benefits including stock options will be commensurate with experience
For information and submitting applications please contact:
The institution I just quit downgraded all of the lab staff positions. A microbiologist with no em experience replaced me with 20 years of em experience only because she lost her job due to downsizing and bumped me out of my position. The management had recently changed (lowered) all of the standards/qualifications so this event could take place. The union was never informed of this move, but that is another story.
A supervisors position should not be part of the union. A smart manager should realize that you cannot hire good help in this field into a supervisor position without at least 7-10 years of EM experience.
The minimum education should be at least a BS in a biological field with at least 5 years experience.
Cheers,
Ed Calomeni
----- Original Message ----- } From: "Dorota Wadowska" {wadowska-at-upei.ca} To: {microscopy-at-sparc5.microscopy.com} Sent: Tuesday, October 08, 2002 10:09 AM
You'll find an entertainingly relevant article at http://www.theatlantic.com/issues/2002/10/working.htm
Libby Shaw
**************************************************************** Elisabeth L. Shaw, Facility Coordinator Surface and Spectroscopy Labs Analytical Shared Experimental Facilities MIT Center for Materials Science and Engineering
Address: MIT Room 13-4149 Tel: 617-253-5045 77 Massachusetts Avenue Email: elshaw-at-mit.edu Cambridge, MA 02139 Fax: 617-258-6478 http://web.mit.edu/cmse/www/ ****************************************************************
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (msandova-at-nd.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, October 10, 2002 at 17:40:13 ---------------------------------------------------------------------------
Email: msandova-at-nd.edu Name: Mayra
Organization: U of Notre Dame
Education: Graduate College
Location: Notre Dame, IN
Question: What is and how does one use a Whipple Hauser optical piece? Thnx!
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sundeep.bhandari-at-nikon.co.uk) from on Thursday, October 10, 2002 at 12:16:30 ---------------------------------------------------------------------------
Question: I am trying to find information about a technique i heard about a few days ago, but can't find any information anywhere on the web. The technique i am researching is 'polarised epi-fluorescence'. I believe you can use it to determine the surface bound orientation of fluorophores; i think the excitation source is linearly polarised and one monitors the fluorescence intensity as a function of polarisation angle.
Any information or pointers in the right direction would be greatly appreciated.
Does anyone have attempted to polish /seal / ? carbon 1/2 " carbon SEM mounts to obtain very smooth surface for HR SEM and be willing to share the procedure? Ultrafine-grain sand paper , carbon colloid, and card-board did not generate a surface which is smooth enough.
The Microscopy Society of America has recently approved the formation of a focused ion beam (FIB) focus interest group (FIG). The function of the FIB FIG will be to organize a symposium for the Microscopy and Microanalysis annual meeting, to hold an annual business meeting of the FIG, and at the discretion of the members, hold a workshop or other FIB sponsored event throughout the year. Individual membership is open to all MSA members at an additional nominal cost of $10 per year above the annual MSA dues commencing with the 2003 yearly MSA dues. A healthy membership within the FIB FIG will ensure that FIB and related topics are an integral part of the M&M meeting. In addition, members of the FIB FIG will have access to workshops and programs designed to further the development and applications of FIB. Vendors are welcome and encouraged to participate in the FIB FIG. Vendors may join the FIB FIG at a nominal cost of $50 per year.
The first FIB FIG workshop is tentatively set for Tuesday March 18, 2003, at the University of Central Florida in Orlando during the joint annual meeting of the FL AVS and Florida Society for Microscopy.
Our goal is to get 50 people to join the FIB FIG within the first year. If you are interested in becoming a member of the FIB FIG and/or interested in attending the 1st FIB FIG Workshop in March, 2003 please contact either:
Joe Michael, Vice Chair, FIB FIG jrmicha-at-sandia.gov
or
Barry Carter, Secretary/Treas, FIB FIG carter-at-cems.umn.edu
******************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor, Mechanical, Materials, and Aerospace Engineering Director, UCF/Cirent Materials Characterization Facility University of Central Florida 12443 Research Parkway, Suite 305 Orlando, FL 32826 email lag-at-mail.ucf.edu phone (407) 882-1500 fax (407) 275-4321 -------------------------------------------------------------------- "Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain ********************************************************************
Th. Bill: What was ~ temp. which you were using? Is that stuff conductive or you have to re-coat it? Marek.
At 07:37 AM 10/11/02 -0400, you wrote: } Apiezon W - a high temp vacuum wax, can be melted on to the surface and } it will be perfectly smooth. } } Bill Miller } } } } At 06:20 PM 10/10/2002 -0700, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Th. Wharton: Please let me know after you try polishing it. We have all possible mills and grinders here, but the pins , which we have, are really tough. Marek.
At 07:22 AM 10/11/02 -0500, you wrote:
} Marek, } } Is this graphitic carbon? If so, you might look into vitreous carbon. This } stuff is really hard, and shiny like a silicon wafer. I was looking for a } really featureless, low Z mount, but not toxic (beryllium), and stumbled on } this. I've just started using it so I haven't tried polishing it yet but I } think it should be considerably easier to polish than graphitic carbon. } } Here's where I got it (sorry, not an EM supplier so you won't get standard } mount geometries here). } } www.atomergic.com } } Regards, } Wharton } } ************************************************************* } Wharton Sinkler, PhD } UOP LLC } 25 E. Algonquin Rd. } Des Plaines, IL 6017-5017 } 847-391-3878 } } } } -----Original Message----- } } From: Marek Malecki, M.D., Ph.D., Professor } } [SMTP:mmalecki-at-facstaff.wisc.edu] } } Sent: Thursday, October 10, 2002 8:21 PM } } To: microscopy-at-sparc5.microscopy.com } } Subject: polishing 1/2 " Cambridge carbon SEM mounts } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Does anyone have attempted to polish /seal / ? carbon 1/2 " carbon SEM } } mounts to obtain very smooth surface for HR SEM and be willing to share } } the } } procedure? Ultrafine-grain sand paper , carbon colloid, and card-board did } } } } not generate a surface which is smooth enough.
I agree with Dr. Sara Miller's point that UA en block staining in some extent helps in preservation of ultrastructure as well as enhancement of the contrast. However, it also has disadvantages such as one has to deal with UA, a biohazard reagent, and sometimes background from crystal precipitation, especially when ethanolic UA is used. More importantly, it is time-consuming because it takes hours for aqueous UA solution to penetrate a sizable tissue block. Therefore, I would say it all depends your tissue and purpose of your analysis.
In my lab, we routinely do en block with samples of blood, tissue culture, virus, bacteria and other microorganisms, which are believed more vulnerable to solvent, but not with solid tissue. We have a lot of samples from kidney biopsy and we didn't find en block made any difference from staining thin sections whereas taking your time to have a sufficient osmication is much more helpful to your structure.
Greg Ning
mohammed y abdulrawoof / f40z006 75760 wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear listers, } Please advise as to the usefulness/advantage of doing enblock staining of } biological tissues as against doing heavy-metal staining of grids with } thin sections. The tissues that I am about to process are for diagnosis } (pathology). } } Thanks } M. Yousuf Abdul-Rawoof
-- Gang Ning, M.D., Ph.D. Director Electron Microscopy Facility Medical College of Wisconsin 8701 Watertown Plank Road Milwaukee, Wisconsin 53226 (414) 456-8344 e-mail: gning-at-mcw.edu http://www.mcw.edu/cellbio/emfacility.html
I have recently acquired a Wild M400 Photomakroskop equipped for Polaroid photography only. I would like to set it up for 35mm photography. I have obtained an appropriate camera and controller, but the necessary relay lens assembly (Wild part # 376 734) is no longer available. So far, I have been unable to find anybody willing to part with one. Has anybody out there had any success improvising a 35mm photography setup on the M400 using other than the Wild original equipment? Any advice would be appreciated.
Ralph Common Michigan State University Division of Human Pathology ralph.common-at-ht.msu.edu
} Dear Mr. Udah; } } I will refer your request to a high powered government official in the } Federal Prosecutor's Office in Newark, New Jersey. } } Sincerely, } } P. Tomic } } -----Original Message----- } } From: ENGR. JOE UDAH [mailto:joeudah123-at-dpr-nig.com] } Sent: Wednesday, October 09, 2002 11:49 AM } To: microscopy-at-sparc5.microscopy.com } Subject: DPR } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
snips fascinating but irrelevant
} further information on the procedures. Feel free to send response by Fax or } TEL; expecting your response urgently. All enquiries should be directed to } the undersigned by FAX OR PHONE. } Looking forward to a good business relationship with you. } } Sincerely, } ENGR. JOE UDAH.
Too late - the Enron management have already got it all stitched up :-)) -- Larry Stoter
We are considering purchasing the latest cryoworkstation (Leica CPC) to plunge freeze samples into liquid propane in preparation for freeze substitution. On our quote there was an automatic filling device listed as optional and we decided not to purchase it to save money (over $6000). Now we are told that it is not optional but required because this unit is not able to be filled manually.
Do any of you use this equipment and know if it can be filled manually or needs an automatic filling device?
See CAPS interspersed below for distinction from the original.
Sara Miller
On Fri, 11 Oct 2002, Gang Ning wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi } } I agree with Dr. Sara Miller's point that UA en block staining in some extent } helps in preservation of ultrastructure as well as enhancement of the } contrast. However, it also has disadvantages such as one has to deal with UA, } a biohazard reagent,
A BIOHAZARD IS A BIOLOGICAL AGENT THAT IS HAZARDOUS (INFECTIOUS). UA IS RADIOACTIVE AND IS A HEAVY METAL. IT, LIKE GLUTARALDEHYDE, OSMIUM, LEAD, RESINS, PROPYLENE OXIDE, ETC., SHOULD BE HANDLED WITH CARE. HANDLED PROPERLY, IT IS NOT ANY MORE DANGEROUS THAN THE OTHER NASTY CHEMICALS ELECTRON MICROSCOPISTS HAVE TO USE. IT IS NOT A BIOLOGICAL AGENT.
and sometimes background from crystal precipitation, } especially when ethanolic UA is used.
WE NEVER SEE PRECIPITATION. PERHAPS TRY DILUTING THE STAIN OR WASHING THOROUGHLY BEFORE USING IT WILL ELIMINATE THE PRECIPITATE. UA IS NOT SOLUBLE IN PHOSPHATE OR CACODYLATE BUFFER; IF YOU DON'T WASH OUT THESE BUFFERS BEFORE ADDING UA IN VERANAL ACETATE BUFFER, UA IN WATER, OR UA IN ETHANOL OR METHANOL, YOU CAN GET PRECIPATE.
More importantly, it is time-consuming } because it takes hours for aqueous UA solution to penetrate a sizable tissue } block.
WE USE 1 MM TISSUE BLOCKS AND STAIN FOR AN HOUR 1% UA IN 0.11M VERONAL ACETATE BUFFER. IF THE TISSUE OR CELL PELLET IS SMALLER, YOU CAN GET AWAY WITH 30 MIN IN UA.
Therefore, I would say it all depends your tissue and purpose of your } analysis. } I AGREE. IT ALL DEPENDS ON YOUR PURPOSE. IF YOU WANT BEAUTIFUL ULTRASTRUCTURE AND YOU'RE NOT LOOKING SPECIFICALLY FOR GLYCOGEN, USE UA EN BLOCK. IF YOU'RE IN A HURRY OR YOU CARE TO SEE ALL THE GLYGOGEN, DON'T USE IT.
} In my lab, we routinely do en block with samples of blood, tissue culture, } virus, bacteria and other microorganisms, which are believed more vulnerable } to solvent, but not with solid tissue. We have a lot of samples from kidney } biopsy and we didn't find en block made any difference from staining thin } sections whereas taking your time to have a sufficient osmication is much } more helpful to your structure. } } Greg Ning } } } } mohammed y abdulrawoof / f40z006 75760 wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Dear listers, } } Please advise as to the usefulness/advantage of doing enblock staining of } } biological tissues as against doing heavy-metal staining of grids with } } thin sections. The tissues that I am about to process are for diagnosis } } (pathology). } } } } Thanks } } M. Yousuf Abdul-Rawoof } } -- } Gang Ning, M.D., Ph.D. } Director } Electron Microscopy Facility } Medical College of Wisconsin } 8701 Watertown Plank Road } Milwaukee, Wisconsin 53226 } (414) 456-8344 } e-mail: gning-at-mcw.edu } http://www.mcw.edu/cellbio/emfacility.html } } } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
} Does anyone have attempted to polish /seal / ? carbon 1/2 " carbon SEM } mounts to obtain very smooth surface for HR SEM and be willing to share the } procedure? Ultrafine-grain sand paper , carbon colloid, and card-board did } not generate a surface which is smooth enough.
We have a user who gets excellent results by gluing a small piece of aluminum foil onto the stub and burnishing it before mounting her micromolluscs. The background is amazingly smooth. I've not tried to duplicate her technique, so I can't claim it will work all the time and with all brands of foil. I have seen machine marks on some foils. I'm also not sure if she uses regular thickness or heavy-duty. I will email her for details when she returns from the field (Tahiti)!
Bug me if you don't hear from me.
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Thanks Tina: Looking forward to the details. We were attempting to polish Al stubs, but they came out not flat enough. Marek.
PS How is your 912 working?
At 03:27 PM 10/11/02 -1000, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello EDX Experts; Would you be willing to share your experience with various EDX mapping programs - in particular deconvolving elemental distribution with corrections for binding and emission energies in alloys or particles embedded in a matrix (C)? Marek.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ggm-at-servidor.unam.mx) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, October 11, 2002 at 11:39:52 ---------------------------------------------------------------------------
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (gbarclay-at-trinidad.net) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, October 11, 2002 at 23:40:22 ---------------------------------------------------------------------------
Question: Light microscope terminology: We all know what a stereomicroscope is and what a compound microscope is, but are they not technically both the same thing? Should not the distinction be between the "simple microscope," basically a highly corrected doublet or triplet lens, forming the Dowland type of microscope, and any kind of microscope with a separate eyepiece and objective lens? If so, then both the stereo and binocular (or monocular) microscopes are compond microscopes. I have been referring to "stereomicroscopes and binocular microscopes" in our Life Sciences Dept, and having some arguments with those who refer "stereomicroscopes and compound microscopes," over what is really what.
Marek Malecki wrote: ================================================================ Does anyone have attempted to polish /seal / ? carbon 1/2 " carbon SEM mounts to obtain very smooth surface for HR SEM and be willing to share the procedure? Ultrafine-grain sand paper , carbon colloid, and card-board did not generate a surface which is smooth enough. ================================================================ Highly Ordered Pyrolytic Graphite (HOPG) which can be freshly cleaved like mica, will cleave to an atomically smooth surface, mirror reflective. The size of the atomically smooth areas depend on the grade of HOPG that is procured. While the cost at first might seem high, remember one can get a number of cleavings out of one HOPG starting "block". This is the substrate often times used by those doing SPM work because of its ultra smooth surface relative to other ways to make smooth nonpolar surfaces in carbon or graphite.
In addition, it is of high purity and other wise behaves like any other form of carbon in terms of its chemical resistance and stability.
More information about HOPG can be found on URL http://www.2spi.com/catalog/new/hopgsub.shtml
Disclaimer: SPI Supplies is a leading supplier of HOPG to microscopy laboratories worldwide so we have a vested interest in seeing more of it used in this kind of application.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
It seems to me that there jas always been one other difference between the two. Most truly binocular (stereo) microscopes do not have an inverted image, whereas most "compound" microscopes have an inverted image.
Ken Converse owner Quality Images third party SEM service Delta, PA
by way of Ask-A-Microscopist wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (gbarclay-at-trinidad.net) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, } October 11, 2002 at 23:40:22 } --------------------------------------------------------------------------- } } } Email: gbarclay-at-trinidad.net } Name: Greg Barclay } } Organization: University of the West Indies } } Education: Graduate College } } Location: St. Augustine, Trinidad, West Indies } } Question: Light microscope terminology: We all know what a } stereomicroscope is and what a compound microscope is, but are they } not technically both the same thing? Should not the distinction be } between the "simple microscope," basically a highly corrected doublet } or triplet lens, forming the Dowland type of microscope, and any kind } of microscope with a separate eyepiece and objective lens? If so, then } both the stereo and binocular (or monocular) microscopes are compond } microscopes. I have been referring to "stereomicroscopes and binocular } microscopes" in our Life Sciences Dept, and having some arguments with } those who refer "stereomicroscopes and compound microscopes," over } what is really what. } } --------------------------------------------------------------------------- } } }
charset="iso-8859-1" Content-Transfer-Encoding: 8bit X-Priority: 3 (Normal) X-MSMail-Priority: Normal X-Mailer: Microsoft Outlook, Build 10.0.2616 Importance: Normal
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id PAA24924 for dist-Microscopy; Sun, 13 Oct 2002 15:21:56 -0500 (CDT) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id PAA24920 for "MicroscopyFilteredEmail2-at-msa.microscopy.com"; Sun, 13 Oct 2002 15:21:26 -0500 (CDT) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id PAA24913 for {Microscopy-at-sparc5.microscopy.com} ; Sun, 13 Oct 2002 15:21:14 -0500 (CDT) Received: from godzilla6.acpub.duke.edu (godzilla6.acpub.duke.edu [152.3.102.8]) by gibson.acpub.duke.edu (8.11.5/8.11.3/Duke-5.0.0) with ESMTP id g9DKFAO11358; Sun, 13 Oct 2002 16:15:10 -0400 (EDT) Received: (from saram-at-localhost) by godzilla6.acpub.duke.edu (8.9.3/8.9.3) id QAA02772; Sun, 13 Oct 2002 16:15:09 -0400 (EDT)
Again, see CAPS below. (CAPS ARE USED TO DIFFERENTIATE COMMENTS, NOT TO CONVEY HOSTILITY.)
On Sun, 13 Oct 2002, gang ning wrote:
} Thanks for the clear definition of biohazard. But it is obvious that } 1) UA is a nasty/hazard chemical. Everyone wants to avoid it if possible,
UA CERTAINLY CAN BE CALLED NASTY. IT DOES REQUIRE SPECIAL HANDLING PROCEDURES, AS DO OTHER CHEMICALS THAT WE USE AND CAN BE HANDLED SAFELY UNDER THE PROPER CONDITIONS. NOT EVERYONE WANTS TO AVOID IT. FOLKS MUST MAKE THEIR OWN JUDGEMENT AS TO WHETHER THE EXTRA PROCESSING TIME AND CARE OF HANDLING ANOTHER CHEMICAL IS WORTH IT TO THEM. NOT USING UA IS FINE. HOWEVER, DO NOT MAKE A GENERALIZATION ABOUT *EVERYONE* JUST BECAUSE YOU CHOOSE NOT TO USE IT.
} 2) it does take at least 1 hr + time for washing thoroughly (how long?) } for a 1 mm3 block .... it is clearly time-consuming procedure
IT DOES REQUIRE EXTRA TIME (1-2 HOURS). ONE CAN STAIN A 1 mm3 BLOCK FOR 30-60 MIN AND WASH FOR 30-60 MIN.
unless your turn around time is not an issue and working overtime is not a } matter with your hospital policy,
TIME IS VERY MUCH AN ISSUE FOR US IN THE HOSPITAL SETTING. OUR TURN AROUND TIME IS *ROUTINELY* NEXT DAY FOR SAMPLES FIXED IN GLUT, OS, UA. TISSUE COMES IN IN THE AM, IS PROCESSED THAT DAY, BAKED OVERNIGHT, CUT AND VIEWED THE NEXT DAY, AND PATHOLOGISTS HAVE MICROGRAPHS *IN HAND* THAT AFTERNOON--ALMOST ALWAYS. MY TECHS RARELY HAVE TO WORK OVERTIME, AND WHEN THEY DO, IT'S USUALLY BECAUSE OF EXTRA CUTTING OF HAVING TO RETRIEVE SPECIMENS OUT OF PARAFFIN BLOCKS OR OFF SLIDES, NOT BECAUSE THEY PUT SOMETHING INTO UA.
YOU ARE CLEARLY VERY NEGATIVE TOWARD THE USE OF UA. FINE. DON'T USE IT. BUT DON'T BE SARCASTIC ABOUT SPECIMEN TURN AROUND TIME AND TECHNICIAN OVERTIME, BECAUSE IT JUST ISN'T AN ISSUE IF SPECIMENS AND WORKLOADS ARE MANAGED PROPERLY.
} 3) en block staining has adversary effect on showing glycogen so is not } suitable for muscle, heart, liver and so on; there is not significant } meaning for kidney .. then what else left? LOTS:
BRAIN, CILIA, TUMORS, VIRUSES, MUSCLE DISEASES OTHER THAN GLYCOGEN STORAGE DISEASES, RESEARCH SAMPLES, ETC.
It is overkill for solid tissue blocks, IT IS NOT OVERKILL WHEN THE BEST PRESERVATION, RATHER THAN DECREASING PROCESSING TIME IS OF IMPORTANCE.
and 4) ethanolic/methanolic UA of higher concentration penetrates tissue } much faster than other aqueous UA solutions and gives fresher image and } better contrast but sometimes leave precipitation around cell member no } matter how one washes before staining.
AS I SAID BEFORE, IF THE CORRECT BUFFER IS USED SO THAT UA DOES NOT PRECIPTATE WITH IT, THERE IS NO PRECIPITATION. PERHAPS YOUR *HIGHER CONCENTRATION* IS A CAUSE OF DIFFICULTY IN WASHING. SINCE UA IS SOLUBLE IN ALCOHOL, UNBOUND, UNPRECIPITATED UA IS EASILY WASHED OUT BY THE DEHYDRATING AGENT(S). IF YOU HAVE PRECIPITATE, IT IS MOST LIKELY DUE TO SOME REACTION WITH THE SOLUTION LEFT IN THE CELL BEFORE YOU ADD THE UA, OR PERHAPS THE *HIGHER* CONCENTRATION YOU USE ISN'T WASHED OUT???
} For EM work, the simpler and better, as long as it works.
YES, THE KEY IS, *AS LONG AS IT WORKS* FOR THE PURPOSE YOU HAVE. IF THE PURPOSE IS TO PRODUCE THE BEST ULTRASTRUCTURE, AND IF MEMBRANES AND OR LIPIDS ARE PART OF THE QUESTION, USE OF UA IS GOOD.
Overkill is always a problem. YES, NO ONE WANTS TO SPEND MORE TIME THAN IS NECESSARY OR USE CHEMICALS THAT CAN BE HAZARDOUS IF NOT HANDLED PROPERLY, BUT UA IS NOT ALWAYS OVERKILL.
IT DEPENDS ON YOUR APPLICATION. EACH TO HIS OWN. } } } } saram-at-duke.edu {mailto:saram-at-duke.edu} wrote: } } } See CAPS interspersed below for distinction from the original. } } } } Sara Miller } } } } On Fri, 11 Oct 2002, Gang Ning wrote: } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com {mailto:ListServer-at-MSA.Microscopy.Com} } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } Hi } } } } } } I agree with Dr. Sara Miller's point that UA en block staining in some extent } } } helps in preservation of ultrastructure as well as enhancement of the } } } contrast. However, it also has disadvantages such as one has to deal with UA, } } } a biohazard reagent, } } } } } } } A BIOHAZARD IS A BIOLOGICAL AGENT THAT IS HAZARDOUS (INFECTIOUS). UA IS } } RADIOACTIVE AND IS A HEAVY METAL. IT, LIKE GLUTARALDEHYDE, OSMIUM, LEAD, } } RESINS, PROPYLENE OXIDE, ETC., SHOULD BE HANDLED WITH CARE. HANDLED } } PROPERLY, IT IS NOT ANY MORE DANGEROUS THAN THE OTHER NASTY CHEMICALS } } ELECTRON MICROSCOPISTS HAVE TO USE. IT IS NOT A BIOLOGICAL AGENT. } } } } and sometimes background from crystal precipitation, } } } } } especially when ethanolic UA is used. } } } } } } } WE NEVER SEE PRECIPITATION. PERHAPS TRY DILUTING THE STAIN OR WASHING } } THOROUGHLY BEFORE USING IT WILL ELIMINATE THE PRECIPITATE. UA IS NOT } } SOLUBLE IN PHOSPHATE OR CACODYLATE BUFFER; IF YOU DON'T WASH OUT THESE } } BUFFERS BEFORE ADDING UA IN VERANAL ACETATE BUFFER, UA IN WATER, OR UA IN } } ETHANOL OR METHANOL, YOU CAN GET PRECIPATE. } } } } More importantly, it is time-consuming } } } } } because it takes hours for aqueous UA solution to penetrate a sizable tissue } } } block. } } } } } } } WE USE 1 MM TISSUE BLOCKS AND STAIN FOR AN HOUR 1% UA IN 0.11M VERONAL } } ACETATE BUFFER. IF THE TISSUE OR CELL PELLET IS SMALLER, YOU CAN GET AWAY } } WITH 30 MIN IN UA. } } } } Therefore, I would say it all depends your tissue and purpose of your } } } } } analysis. } } } } } I AGREE. IT ALL DEPENDS ON YOUR PURPOSE. IF YOU WANT BEAUTIFUL } } ULTRASTRUCTURE AND YOU'RE NOT LOOKING SPECIFICALLY FOR GLYCOGEN, USE UA EN } } BLOCK. IF YOU'RE IN A HURRY OR YOU CARE TO SEE ALL THE GLYGOGEN, DON'T } } USE IT. } } } } } In my lab, we routinely do en block with samples of blood, tissue culture, } } } virus, bacteria and other microorganisms, which are believed more vulnerable } } } to solvent, but not with solid tissue. We have a lot of samples from kidney } } } biopsy and we didn't find en block made any difference from staining thin } } } sections whereas taking your time to have a sufficient osmication is much } } } more helpful to your structure. } } } } } } Greg Ning } } } } } } } } } } } } mohammed y abdulrawoof / f40z006 75760 wrote: } } } } } } } ------------------------------------------------------------------------ } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com {mailto:ListServer-at-MSA.Microscopy.Com} } } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } -----------------------------------------------------------------------. } } } } } } } } Dear listers, } } } } Please advise as to the usefulness/advantage of doing enblock staining of } } } } biological tissues as against doing heavy-metal staining of grids with } } } } thin sections. The tissues that I am about to process are for diagnosis } } } } (pathology). } } } } } } } } Thanks } } } } M. Yousuf Abdul-Rawoof } } } } } } } -- } } } Gang Ning, M.D., Ph.D. } } } Director } } } Electron Microscopy Facility } } } Medical College of Wisconsin } } } 8701 Watertown Plank Road } } } Milwaukee, Wisconsin 53226 } } } (414) 456-8344 } } } e-mail: gning-at-mcw.edu {mailto:gning-at-mcw.edu} } } } http://www.mcw.edu/cellbio/emfacility.html } } } } } } } } } } } } } } } } } } } Sara E. Miller, Ph. D. } } P. O. Box 3712 } } Duke University Medical Center } } Durham, NC 27710 } } Ph: 919 684-3452 } } FAX: 919 684-3265 } } } } } } -- } } Gang Ning, M.D., Ph.D. } } Director } } Electron Microscopy Facility } } Medical College of Wisconsin } } 8701 Watertown Plank Road } } Milwaukee, WI 53226 } } (414) 456-8141 } } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Another option is to glue a small round glass coverslip onto the stub with carbon paste, then coat with gold, then attach specimens, re-coat.
} Hi, Marek- } } } Does anyone have attempted to polish /seal / ? carbon 1/2 " carbon SEM } } mounts to obtain very smooth surface for HR SEM and be willing to share the } } procedure? Ultrafine-grain sand paper , carbon colloid, and card-board did } } not generate a surface which is smooth enough. } } We have a user who gets excellent results by gluing a small piece of } aluminum foil onto the stub and burnishing it before mounting her } micromolluscs. The background is amazingly smooth. I've not tried to } duplicate her technique, so I can't claim it will work all the time and } with all brands of foil. I have seen machine marks on some foils. I'm also } not sure if she uses regular thickness or heavy-duty. I will email her for } details when she returns from the field (Tahiti)! } } Bug me if you don't hear from me. } } Aloha, } Tina } } } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } ****************************************************************************
We have a preference for mounting a coated 0.5" coverslip onto the stub with colloidal dag. The cover slip is coated with poly-L-lysine either before or after mounting on the stub. This holds the sample well and gives a smooth black background after sputter coating. Regards JVN
Tina Carvalho wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, Marek- } } } } Does anyone have attempted to polish /seal / ? carbon 1/2 " carbon SEM } } mounts to obtain very smooth surface for HR SEM and be willing to share the } } procedure? Ultrafine-grain sand paper , carbon colloid, and card-board did } } not generate a surface which is smooth enough. } } } } We have a user who gets excellent results by gluing a small piece of } aluminum foil onto the stub and burnishing it before mounting her } micromolluscs. The background is amazingly smooth. I've not tried to } duplicate her technique, so I can't claim it will work all the time and } with all brands of foil. I have seen machine marks on some foils. I'm also } not sure if she uses regular thickness or heavy-duty. I will email her for } details when she returns from the field (Tahiti)! } } Bug me if you don't hear from me. } } Aloha, } Tina } } } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } **************************************************************************** } } } } }
-- John V Nailon Executive Officer and Operations Manager The Centre for Microscopy and Microanlaysis The University of Queensland St Lucia QLD 4072 Tel: +61-7-33654214 Fax: +61-7-33654422 WWW: http://www.uq.edu.au/nanoworld
Dear Listers, I´m searching for a smooth muscle anti-Actin antibody (recognizing mouse smooth muscle) which also works on formaldehyde fixed tissue sections. Any hints or recommendations are welcome.
I must jump in on this one. A real "stereo" microscope has two parallel optical paths. That is, two objective lenses. No microscope with one objective lens can be "stereo." Albeit there may be two eyepieces but without two separate optical paths the images in both eyepieces must necessarily be identical. It's like listening to stereo sound but if the left and right channels are identical, hence no depth or direction to the sound.
Inverted or non-inverted images do not define a compound microscope since many are made with an "image erector" which is simply an additional lens to reverse the image so you view the sample as it is with the unaided eye. Most "micromanipulator" systems have image erectors on them for that purpose. It helps one to not have to re-wire one's neurons to interpret left as right when having to use hand manipulation under the microscope. I have found that wiring and re-wiring neurons problematic.
Peter Anadigics, Inc.
-----Original Message----- } From: qualityimages [mailto:qualityimages-at-netrax.net] Sent: Saturday, October 12, 2002 6:25 PM To: by way of Ask-A-Microscopist; Microscopy
It seems to me that there jas always been one other difference between the two. Most truly binocular (stereo) microscopes do not have an inverted image, whereas most "compound" microscopes have an inverted image.
Ken Converse owner Quality Images third party SEM service Delta, PA
by way of Ask-A-Microscopist wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (gbarclay-at-trinidad.net) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, } October 11, 2002 at 23:40:22 } --------------------------------------------------------------------------- } } } Email: gbarclay-at-trinidad.net } Name: Greg Barclay } } Organization: University of the West Indies } } Education: Graduate College } } Location: St. Augustine, Trinidad, West Indies } } Question: Light microscope terminology: We all know what a } stereomicroscope is and what a compound microscope is, but are they } not technically both the same thing? Should not the distinction be } between the "simple microscope," basically a highly corrected doublet } or triplet lens, forming the Dowland type of microscope, and any kind } of microscope with a separate eyepiece and objective lens? If so, then } both the stereo and binocular (or monocular) microscopes are compond } microscopes. I have been referring to "stereomicroscopes and binocular } microscopes" in our Life Sciences Dept, and having some arguments with } those who refer "stereomicroscopes and compound microscopes," over } what is really what. } } --------------------------------------------------------------------------- } } }
} } } } } Hi, } } } } I usually don't weigh in on this type of thing but, as a past negotiator } } for the teachers in my old school system and as an ardent advocate of the } } importance of experience in microscopy, here is my two cents: } } 1. As a negotiator - I worked very closely with both the Mass Teachers } } Association (MTA) and the National Teachers Association. Ironically, our } } "supervisors" (curriculum supervisors, principals and vice principals) } } needed the protection of our association (MTA and NEA preferred not to be } } called unions but they filled the role of protectors and negotiators), so, } } to offer them that support, we opened membership to them but, for } } negotiation purposes, separated them into their own negotiating } group. The } } position of lab director } } } } 2. As for your 20 years' experience: I earn my living training } } microscopists. I head a group of consultants who specialize in } } training. I am also a contributing editor to two key publications. Our } } company is THE primary source of market research in the microscopy and } } imaging industry. With well over 20 year's experience in the field as a } } microscopist, a trainer, and now, a recognized industry watcher, I will } } tell you the following: } } The general rule of thumb is that it takes, on average, at least a year } for } } a microscopist to become proficient. Why? } } a.Unlike any other analytical discipline, microscopy demands a high level } } of eye-brain-hand coordination. } } b. Unlike most other disciplines, we don't have one or two major pieces of } } instrumentation to master; we have multiple instrument systems. We } have to } } know how to prepare our samples, which type of microscopy to use, how to } } interpret the myriad types of images each type of microscopy can produce, } } and, finally, how to integrate a complex electronic system including } } cameras, computers, and software, to collect, archive, process, and } measure } } our images. } } c. Unlike most other disciplines, there is no replacement for the mental } } library of images which a microscopist collects with experience. Yes, a } } spectroscopist will learn the nuances of an upfield or downfield shift } } characteristic of a specific spectrum, but no other discipline requires } the } } mental collection and correlation of image information like microscopy. } } } } As a master teacher and contributing editor, I know that I must weigh my } } words carefully. However, as you can tell, I feel very strongly about the } } erosion of microscopy experience which inevitably accompanies budget } } cuts. Please feel free to quote me to your management on any or all of } } these comments. } } } } Best regards... and strong support.... } } Barbara Foster } } Microscopy/Microscopy Education } } 125 Paridon Street, Suite 102 } } Springfield, MA 01118 } } PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com } } } } } } } } } } At 06:43 PM 10/10/02 -0400, Ed Calomeni wrote: } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } Hi Dorota, } } } } } } Fight this with all you have. } } } } } } The institution I just quit downgraded all of the lab staff positions. A } } } microbiologist with no em experience replaced me with 20 years of em } } } experience only because she lost her job due to downsizing and bumped } me out } } } of my position. The management had recently changed (lowered) all of the } } } standards/qualifications so this event could take place. The union was } } } never informed of this move, but that is another story. } } } } } } A supervisors position should not be part of the union. A smart manager } } } should realize that you cannot hire good help in this field into a } } } supervisor position without at least 7-10 years of EM experience. } } } } } } The minimum education should be at least a BS in a biological field } with at } } } least 5 years experience. } } } } } } Cheers, } } } } } } Ed Calomeni } } } } } } } } } } } } ----- Original Message ----- } } } } From: "Dorota Wadowska" {wadowska-at-upei.ca} } } } To: {microscopy-at-sparc5.microscopy.com} } } } Sent: Tuesday, October 08, 2002 10:09 AM } } } Subject: EM job classificcation } } } } } } } } } } } ------------------------------------------------------------------------ } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } -----------------------------------------------------------------------. } } } } } } } } } } } } Hi Listservers! } } } } I have a few questions that I would like to ask you If you feel you } } } } know the answer please reply. My university together with local } } } } union reclassified staff positions according to Aiken plan. Needless } } } } to say my position was downgraded. } } } } My questions are: } } } } 1. Can EM supervisory position be compared to any on campus } } } } staff positions (university has Department of Sciences and } } } } Veterinary Medecine)? I supervise small multi--user lab, we have 2 } } } } TEM, and we do research, diagnostics, teaching (under- and } } } } graduate level) in the field of biological sciences. No fancy staff } like } } } } X-ray analysis or cryo. } } } } 2. What in your opinion is a necessary minimum experience } } } } required to perform this job ? I am talking about accepting the } } } } position and immediately getting into the stream of work. } } } } 3. What in your opinion is the minimum level of education that is } } } } required to perform this job? } } } } } } } } You can reply directly to me. Thanks in advance } } } } Frustrated } } } } Dorota } } } } } }
Just because a microscope has two eyepieces, does not mean that it is a "stereo" microscope.
A stereo microscope has not only two eyepieces, but also two objectives, and two separate light paths. Because of having two slightly offset images, you see a three dimensional image. Usually these are lower powered systems, but still compound microscopes (multiple lens systems).
On a high power (relatively) microscope with a binocular head, you only have one objective, and one light path from the specimen to the head. The prism(s) in the head divide the light path to provide multiple images, but they are all (three in the case of an extra camera port) identical. This does not provide a 3-D image, it just makes it easier on the eyes and brain to process the image because one eye is not distracted.
I am not a scientist, and would be interested in any corrections to the above.
John W. Raffensperger, Jr. IS Manager Helwig Carbon Products, Inc.
Question: Light microscope terminology: We all know what a stereomicroscope is and what a compound microscope is, but are they not technically both the same thing?
I need to prepare TEM cross-section samples of alternate SiO2 and TiO2 layers on a Si substrate. For that purpose I would appreciate suggestions on glues, suitable for TEM work, that have a sputtering rate not too different from my non homogeneous layers. The final sputtering will be on an Ion Mill (4keV) using Ar+ ions. All suggestions are welcome (including vendors) either to the list or to me directly. Thank you,
Isabel
Isabel Nogueira Instituto Superior Técnico Dep. Materiais Avenida Rovisco Pais 1049-001 Lisboa Portugal tel.: +351 218418123 fax: +351 218418120 email: isabeln-at-popsrv.ist.utl.pt
} From: Barbara Foster {bfoster-at-mme1.com} } Subject: Re: EM job classification } } } } } } } } } Hi, } } } } } } I usually don't weigh in on this type of thing but, as a past negotiator } } } for the teachers in my old school system and as an ardent advocate of the } } } importance of experience in microscopy, here is my two cents: } } } 1. As a negotiator - I worked very closely with both the Mass Teachers } } } Association (MTA) and the National Teachers Association. Ironically, our } } } "supervisors" (curriculum supervisors, principals and vice principals) } } } needed the protection of our association (MTA and NEA preferred not to be } } } called unions but they filled the role of protectors and negotiators), } } so, } } } to offer them that support, we opened membership to them but, for } } } negotiation purposes, separated them into their own negotiating } } group. The } } } position of lab director } } } } } } 2. As for your 20 years' experience: I earn my living training } } } microscopists. I head a group of consultants who specialize in } } } training. I am also a contributing editor to two key publications. Our } } } company is THE primary source of market research in the microscopy and } } } imaging industry. With well over 20 year's experience in the field as a } } } microscopist, a trainer, and now, a recognized industry watcher, I will } } } tell you the following: } } } The general rule of thumb is that it takes, on average, at least a } } year for } } } a microscopist to become proficient. Why? } } } a.Unlike any other analytical discipline, microscopy demands a high level } } } of eye-brain-hand coordination. } } } b. Unlike most other disciplines, we don't have one or two major } } pieces of } } } instrumentation to master; we have multiple instrument systems. We } } have to } } } know how to prepare our samples, which type of microscopy to use, how to } } } interpret the myriad types of images each type of microscopy can produce, } } } and, finally, how to integrate a complex electronic system including } } } cameras, computers, and software, to collect, archive, process, and } } measure } } } our images. } } } c. Unlike most other disciplines, there is no replacement for the mental } } } library of images which a microscopist collects with experience. Yes, a } } } spectroscopist will learn the nuances of an upfield or downfield shift } } } characteristic of a specific spectrum, but no other discipline } } requires the } } } mental collection and correlation of image information like microscopy. } } } } } } As a master teacher and contributing editor, I know that I must weigh my } } } words carefully. However, as you can tell, I feel very strongly about } } the } } } erosion of microscopy experience which inevitably accompanies budget } } } cuts. Please feel free to quote me to your management on any or all of } } } these comments. } } } } } } Best regards... and strong support.... } } } Barbara Foster } } } Microscopy/Microscopy Education } } } 125 Paridon Street, Suite 102 } } } Springfield, MA 01118 } } } PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com } } } } } } } } } } } } } } } At 06:43 PM 10/10/02 -0400, Ed Calomeni wrote: } } } } ------------------------------------------------------------------------ } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } -----------------------------------------------------------------------. } } } } } } } } } } } } Hi Dorota, } } } } } } } } Fight this with all you have. } } } } } } } } The institution I just quit downgraded all of the lab staff positions. A } } } } microbiologist with no em experience replaced me with 20 years of em } } } } experience only because she lost her job due to downsizing and bumped } } me out } } } } of my position. The management had recently changed (lowered) all of the } } } } standards/qualifications so this event could take place. The union was } } } } never informed of this move, but that is another story. } } } } } } } } A supervisors position should not be part of the union. A smart manager } } } } should realize that you cannot hire good help in this field into a } } } } supervisor position without at least 7-10 years of EM experience. } } } } } } } } The minimum education should be at least a BS in a biological field } } with at } } } } least 5 years experience. } } } } } } } } Cheers, } } } } } } } } Ed Calomeni } } } } } } } } } } } } } } } } ----- Original Message ----- } } } } } From: "Dorota Wadowska" {wadowska-at-upei.ca} } } } } To: {microscopy-at-sparc5.microscopy.com} } } } } Sent: Tuesday, October 08, 2002 10:09 AM } } } } Subject: EM job classificcation } } } } } } } } } } } } } } } ------------------------------------------------------------------------ } } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America } } } } } To Subscribe/Unsubscribe -- Send Email to } } ListServer-at-MSA.Microscopy.Com } } } } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } } -----------------------------------------------------------------------. } } } } } } } } } } } } } } } Hi Listservers! } } } } } I have a few questions that I would like to ask you If you feel you } } } } } know the answer please reply. My university together with local } } } } } union reclassified staff positions according to Aiken plan. Needless } } } } } to say my position was downgraded. } } } } } My questions are: } } } } } 1. Can EM supervisory position be compared to any on campus } } } } } staff positions (university has Department of Sciences and } } } } } Veterinary Medecine)? I supervise small multi--user lab, we have 2 } } } } } TEM, and we do research, diagnostics, teaching (under- and } } } } } graduate level) in the field of biological sciences. No fancy } } staff like } } } } } X-ray analysis or cryo. } } } } } 2. What in your opinion is a necessary minimum experience } } } } } required to perform this job ? I am talking about accepting the } } } } } position and immediately getting into the stream of work. } } } } } 3. What in your opinion is the minimum level of education that is } } } } } required to perform this job? } } } } } } } } } } You can reply directly to me. Thanks in advance } } } } } Frustrated } } } } } Dorota } } } } } } } }
I think, Peter and you are right with the statement that different light paths are necessary to achive a true stereo effect. I am not so sure, that 2 separate objectives are really required. I know of at least one company (I have no affiliation with or interest in that company) that makes a 3D microscope and I think they are using only one objective lens (http://www.edge-3d.com).
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: John W. Raffensperger, Jr. [mailto:johnr-at-helwigcp.com] Sent: Monday, October 14, 2002 8:59 AM To: 'by way of Ask-A-Microscopist'; Microscopy-at-sparc5.microscopy.com
Just because a microscope has two eyepieces, does not mean that it is a "stereo" microscope.
A stereo microscope has not only two eyepieces, but also two objectives, and two separate light paths. Because of having two slightly offset images, you see a three dimensional image. Usually these are lower powered systems, but still compound microscopes (multiple lens systems).
On a high power (relatively) microscope with a binocular head, you only have one objective, and one light path from the specimen to the head. The prism(s) in the head divide the light path to provide multiple images, but they are all (three in the case of an extra camera port) identical. This does not provide a 3-D image, it just makes it easier on the eyes and brain to process the image because one eye is not distracted.
I am not a scientist, and would be interested in any corrections to the above.
John W. Raffensperger, Jr. IS Manager Helwig Carbon Products, Inc.
Question: Light microscope terminology: We all know what a stereomicroscope is and what a compound microscope is, but are they not technically both the same thing?
1) Swedish Biopharma at Large - What Next? 2) New Report: 'The Next Generation Antifungals: Triazoles vs. Peptides' 3) Recently published BioSeeker Reports 4) Swedish Biotech Packages 5) Seek And You Will Find 6) Coming reports: HCV Therapy / Winning the Osteoporosis Therapy Race 7) Subscription 8) BioSeeker Online Store
1) SWEDISH BIOPHARMA INDUSTRY AT LARGE - WHAT NEXT?
The Swedish biopharma industry has more than doubled in the last six years, growing from 25 companies in 1996 to the current 52. Next to the medical device industry the biopharma industry is the largest biotech industry segment in Sweden. The predominant disease groups for R&D in Swedish biopharma are: Inflammation / autoimmune, metabolic, cancer, infectious, and neurological diseases. However, the pipeline of the industry is in its infancy.
Read the full version of this free, analytical BioNewsletter article at http://www.bioseeker.com/docs/topics/topic_swepharma.asp
2) THE NEXT GENERATION ANTIFUNGALS : TRIAZOLES VS PEPTIDES - A New Report from BioSeeker Group
According to the Antifungal Work Group at NIH there is a "critical need for the development of antifungal agents". There will be tight competition for market shares when PfizerÕs patent on fluconazole (the active ingredient in Diflucan(TM)) expires in 2004. Both Bristol-Myers Squibb and Schering-Plough will try to compete for market shares. In this Highlight Report, we present new data on activites in the field of triazole and peptide antifungals. Topics considered in this report are :
- Research (spectrums, activities, side-effects, resistance and drug interactions) - Patents - Clinical trials (indications, status and activities) - Partnerships - Expected market launches and sales
For more information about this report, please visit http://www.bioseeker.com/index.asp?report=antifungal
a) Order the new report 'Swedish Biopharma Industry' together with the report 'Swedish Biotechnology Industry' (published Jan. 2001) for $1050 (save $200).
b) Obtain five free company profiles from the Swedish Biotech Corporate Index with any purchase of 'Swedish Biopharma Industry' (price $850 US).
c) Order 'Swedish Biotechnology Industry' together with the address list 'Complete Contact Data to Swedish Biotechnology Companies', updated continously and containing contact details to more than 320 Swedish Biotech companies, for $699 (save $350).
When ordering one of these packages, please place your order by fax and use reference codes SWEPACK-A, B or C. (Download orderform at www.bioseeker.com/docs/orderForm.pdf)
Identify and locate your potential customers, competitors, and partners by using BioSeeker Group's world-wide company search service. This service is based on BioSeeker's in-house company database built up during four years of operation and containing more than 10,000 life science companies globally.
We offer you this service in 3 steps:
Step 1. Search & Identify (399 USD)
Step 2. Locate & Validate (699 USD)
Step 3. Select & Build (individual price). Choose the companies of interest and BSG builds extensive profiles of those.
For more information about this service, please contact us at bioinfo-at-bioseeker.com
Treatment of HCV (Hepatitis C Virus) is a market with unmet medical needs, with relatively young people infected and a high risk for chronic disease. The coming decade will see exciting developments in HCV therapy thanks to breakthroughs in overcoming a major barrier to development: the lack of dedicated models suitable for validating candidate compounds in a low-cost, highthroughput format. BioSeeker Group has identified at least 115 companies with interest in various stages in the field of HCV therapy. Out of these 100, approximately 10 of the 'big pharma' are represented. Our coming report "Molecular Approaches Towards New HCV Therapy" analyses the field of new HCV therapeutics and assess possible outcomes for drugs and companies in this sector.
Please contact us at bioinfo-at-bioseeker.com for more information about this report.For further reading about HCV, please visit our free analytical article on this topic (http://www.bioseeker.com/docs/topics/topic_hcv.asp)
- Winning the Osteoporosis Therapy Race
Osteoporosis, deterioration of bone tissue and an increased risk of fractures, is considered by the World Health Organization to be second only to cardiovascular disease as a leading healthcare problem. According to other studies, sales of osteoporosis drugs in these markets totaled $3.4 billion in 2000, and are expected to reach $6.5 billion by 2005. A new class of osteoporosis therapies, designed to build bone mass in contrast to currently available anti-osteoporosis medicines are presently under study. These anabolic therapies consist of parathyroid hormone (FortÎoª and ALX-11) or fluoride to stimulate bone formation, but their safety and efficacy remain to be established. The first of these drugs to reach the market will probably be FortÎoª, known generically as teriparatide (recombinant parathyroid hormone).
Please contact us for more information about this report (bioinfo-at-bioseeker.com). For further reading about osteoporosis, please visit our free analytical article on this topic (http://www.bioseeker.com/docs/topics/topic_osteoporosis.asp)
ADVERTISING Do you have a message that you would like the biotech community to know? BioSeeker Group's Newsletter targets managers, scientists, venture capitalists, consultants and many more world wide. Contact us at sales-at-bioseeker.com for further information on how to advertise in BSG BioNewsletter.
COURTESY
If you would like to comment on our newsletter please send us an email to feedback-at-bioseeker.com
If you would like to be removed from our newsletter list, please reply to unsubscribe-at-bioseeker.com
Isabel, There are two epoxy glues that work well. Epo-Tek 353ND from Epoxy Technology is one. Gatan's G1 is equivalent to this. The other is M-bond 610. I like the Epo-Tek 353ND; mostly because it has a longer shelf life, but also because I believe that it is stronger. I believe that you can get thinner glue joints with the M-bond 610, though.
BTW, Your system is ideal for using the small angle cleavage technique. You can forgo glueing, dimpling, and ion milling altogether and make about 10 samples per hour. Check out the South Bay Technology web site and look at the Microcleave kit and examples that they have. A detailed description of the technique has been written up by John McCaffrey and myself in the number 4 proceedings book on TEM sample preparation from the MRS. John also has the original publication in the number 3 book from MRS. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
-----Original Message----- } From: Isabel Nogueira [mailto:isabeln-at-popsrv.ist.utl.pt] Sent: Monday, October 14, 2002 12:16 PM To: Microscopy-at-sparc5.microscopy.com
Hello,
I need to prepare TEM cross-section samples of alternate SiO2 and TiO2 layers on a Si substrate. For that purpose I would appreciate suggestions on glues, suitable for TEM work, that have a sputtering rate not too different from my non homogeneous layers. The final sputtering will be on an Ion Mill (4keV) using Ar+ ions. All suggestions are welcome (including vendors) either to the list or to me directly. Thank you,
Isabel
Isabel Nogueira Instituto Superior Técnico Dep. Materiais Avenida Rovisco Pais 1049-001 Lisboa Portugal tel.: +351 218418123 fax: +351 218418120 email: isabeln-at-popsrv.ist.utl.pt
Isabel Nogueira wrote: ========================================================= I need to prepare TEM cross-section samples of alternate SiO2 and TiO2 layers on a Si substrate. For that purpose I would appreciate suggestions on glues, suitable for TEM work, that have a sputtering rate not too different from my non homogeneous layers. The final sputtering will be on an Ion Mill (4keV) using Ar+ ions. All suggestions are welcome (including vendors) either to the list or to me directly. =========================================================== Several different glues have been mentioned on the listserver, one being M- Bond™ 610, which is described on URL http://www.2spi.com/catalog/spec_prep/glue.shtml
Disclaimer: SPI Supplies is a worldwide distributor for the M-Bond 610 product.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 USA
TEL: +1-949-492-2600 Toll-free in the USA: +1-800-728-2233 FAX: +1-949-492-1499
email: henriks-at-southbaytech.com
Celebrating 38 years of providing Materials Processing Solutions for Metallogaphy, Crystallography and Electron Microscopy.
Please visit us online at www.southbaytech.com. Isabel Nogueira wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello, } } I need to prepare TEM cross-section samples of alternate SiO2 and TiO2 } layers on a Si substrate. For that purpose I would appreciate suggestions on } glues, suitable for TEM work, that have a sputtering rate not too different } from my non homogeneous layers. The final sputtering will be on an Ion Mill } (4keV) using Ar+ ions. } All suggestions are welcome (including vendors) either to the list or to me } directly. } Thank you, } } Isabel } } Isabel Nogueira } Instituto Superior Técnico } Dep. Materiais } Avenida Rovisco Pais } 1049-001 Lisboa } Portugal } tel.: +351 218418123 } fax: +351 218418120 } email: isabeln-at-popsrv.ist.utl.pt
The information contained in this message and any attachments is privileged and confidential. This message is intended for the individual or entity addressed. If you are not the intended recipient, please do not read, copy or disclose this communication. Notify the sender of the mistake by calling +1-949-492-2600 and delete this message from your system.
Almost all modern stereo microscopes use a single objective lens that have two light paths of varying complexity viewing the specimen at different view points through a single objective.
The light paths are not parallel but view the subject from a slightly different angle and present them to the eye at an angle compatible with the brains processing methods to make it appear as a 3D image. How the light paths are handled between the subject and the eye may have them at any relation to each other but when they reach the eye they are at an angle that make them appear to be 3D images.
Viewing a subject at an angle though a common objective introduces some distortion but less than viewing the same subject through two completely separate microscopes that are an angle to the subject.
If you have doubts about this find a AO Cycloptic with misaligned lenses in the cylinder that change the magnification and see some of the weird ways an image can behave when it is presented to the eyes incorrectly.
Gordon
Gordon Couger Stillwater, OK www.couger.com/gcouger
} From: "Peter Tomic" {PTomic-at-anadigics.com} : : I must jump in on this one. A real "stereo" microscope has two parallel : optical paths. That is, two objective lenses. No microscope with one : objective lens can be "stereo." Albeit there may be two eyepieces but : without two separate optical paths the images in both eyepieces must : necessarily be identical. It's like listening to stereo sound but if the : left and right channels are identical, hence no depth or direction to the : sound. : : Inverted or non-inverted images do not define a compound microscope since : many are made with an "image erector" which is simply an additional lens to : reverse the image so you view the sample as it is with the unaided eye. : Most "micromanipulator" systems have image erectors on them for that : purpose. It helps one to not have to re-wire one's neurons to interpret : left as right when having to use hand manipulation under the microscope. I : have found that wiring and re-wiring neurons problematic. : : Peter : Anadigics, Inc. : : -----Original Message----- : } From: qualityimages [mailto:qualityimages-at-netrax.net] : Sent: Saturday, October 12, 2002 6:25 PM : To: by way of Ask-A-Microscopist; Microscopy : Subject: Re: Ask-A-Microscopist:LM terminology : : : ------------------------------------------------------------------------ : The Microscopy ListServer -- Sponsor: The Microscopy Society of America : To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com : On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html : -----------------------------------------------------------------------. : : : It seems to me that there jas always been one other difference between : the two. Most truly binocular (stereo) microscopes do not have an : inverted image, whereas most "compound" microscopes have an inverted image. : : Ken Converse : owner : Quality Images : third party SEM service : Delta, PA : : by way of Ask-A-Microscopist wrote: : : } ------------------------------------------------------------------------ : } The Microscopy ListServer -- Sponsor: The Microscopy Society of America : } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com : } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html : } -----------------------------------------------------------------------. : } : } : } Below is the result of your feedback form (NJZFM-ultra-55). It was : } submitted by (gbarclay-at-trinidad.net) from : } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, : } October 11, 2002 at 23:40:22 : } : -------------------------------------------------------------------------- - : } : } : } Email: gbarclay-at-trinidad.net : } Name: Greg Barclay : } : } Organization: University of the West Indies : } : } Education: Graduate College : } : } Location: St. Augustine, Trinidad, West Indies : } : } Question: Light microscope terminology: We all know what a : } stereomicroscope is and what a compound microscope is, but are they : } not technically both the same thing? Should not the distinction be : } between the "simple microscope," basically a highly corrected doublet : } or triplet lens, forming the Dowland type of microscope, and any kind : } of microscope with a separate eyepiece and objective lens? If so, then : } both the stereo and binocular (or monocular) microscopes are compond : } microscopes. I have been referring to "stereomicroscopes and binocular : } microscopes" in our Life Sciences Dept, and having some arguments with : } those who refer "stereomicroscopes and compound microscopes," over : } what is really what. : } : } : -------------------------------------------------------------------------- - : } : } : } : : : : : :
I agree, my Lynx is stereo with only one objective.
http://www.visioneng.com/
Jim
} From Microscopy-request-at-sparc5.microscopy.com Tue Oct 15 03:54:02 2002 } From: Mike Bode {mb-at-Soft-Imaging.com} } To: "'Microscopy-at-MSA.Microscopy.Com'" {Microscopy-at-sparc5.microscopy.com} } Subject: RE: Ask-A-Microscopist:LM terminology } Date: Mon, 14 Oct 2002 14:35:58 -0600 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I think, Peter and you are right with the statement that different light } paths are necessary to achive a true stereo effect. I am not so sure, that 2 } separate objectives are really required. I know of at least one company (I } have no affiliation with or interest in that company) that makes a 3D } microscope and I think they are using only one objective lens } (http://www.edge-3d.com). } } mike } } } Michael Bode, Ph.D. } Soft Imaging System Corp. } 12596 West Bayaud Avenue } Suite 300 } Lakewood, CO 80228 } =================================== } phone: (888) FIND SIS } (303) 234-9270 } fax: (303) 234-9271 } email: mailto:info-at-soft-imaging.com } web: http://www.soft-imaging.com } =================================== } } } } -----Original Message----- } } From: John W. Raffensperger, Jr. [mailto:johnr-at-helwigcp.com] } Sent: Monday, October 14, 2002 8:59 AM } To: 'by way of Ask-A-Microscopist'; Microscopy-at-sparc5.microscopy.com } Subject: RE: Ask-A-Microscopist:LM terminology } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Just because a microscope has two eyepieces, does not mean that it is a } "stereo" microscope. } } A stereo microscope has not only two eyepieces, but also two objectives, } and two separate light paths. Because of having two slightly offset } images, you see a three dimensional image. Usually these are lower } powered systems, but still compound microscopes (multiple lens systems). } } On a high power (relatively) microscope with a binocular head, you only } have one objective, and one light path from the specimen to the head. } The prism(s) in the head divide the light path to provide multiple } images, but they are all (three in the case of an extra camera port) } identical. This does not provide a 3-D image, it just makes it easier } on the eyes and brain to process the image because one eye is not } distracted. } } I am not a scientist, and would be interested in any corrections to the } above. } } John W. Raffensperger, Jr. } IS Manager } Helwig Carbon Products, Inc. } } } Email: gbarclay-at-trinidad.net } Name: Greg Barclay } } Organization: University of the West Indies } } Education: Graduate College } } Location: St. Augustine, Trinidad, West Indies } } Question: Light microscope terminology: We all know what a } stereomicroscope is and what a compound microscope is, but are they } not technically both the same thing? } }
To enhance contrast a bit we kept stock bottles of 50% and 70% etoh with 1-2% UA adding contrast while dehydrating. As long as tissue pieces were small this worked well. Solution was drawn from the top or filtered to prevent the precip that collected in the bottom. Time was doubled in these two steps. Obviously if extraction or glycogen was important then this was not done.
Scott Whittaker Laboratories of Analytical Biology Smithsonian Institution National Museum of Natural History PO Box 37012 MRC104 Washington DC 20013-7012 202-357-1651
} } } mohammed y abdulrawoof / f40z006 75760 {mdyousuf-at-KSU.EDU.SA} 10/10/02 12:35PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear listers, Please advise as to the usefulness/advantage of doing enblock staining of biological tissues as against doing heavy-metal staining of grids with thin sections. The tissues that I am about to process are for diagnosis (pathology).
ESEM images of bacteria are different from gold coated images of either air dried or CPD bacteria.
Firstly many bacteria produce slime. In air dried samples this collapses. In CPD samples the solvents probably remove it or condense it. In ESEM the slime is retained so that one often does not see naked bacteria at all. This conflict with expectation can lead to disappointment!
Secondly, gold gives a good SE yield from the surface. In ESEM (tungsten filament) bacteria yield less SE and from a greater depth. This gives a "softer" image of bacteria than one is used to.
Don't worry about coating units being outmoded. We showed school visitors 7 samples today. All were gold coated! The samples are easier to image.
With many dry materials samples it is easier to gold coat and then image in ESEM. I know this is cheating ....
We are still using CPD for projects comparing preparation methods.
Dave
On Thu, 10 Oct 2002 12:40:14 -0500 "Garber, Charles A." {cgarber-at-2spi.com} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } David Patton wrote: } ========================================================== } Disclaimer: ESEM images of bacteria have a different quality to those that } } have been dried and gold coated; so warn your user to avoid } disappointment. } =========================================================== } } Are you comparing ESEM images with a) air dried and gold coated or b) } critical point dried and gold coated? } } It would be obvious that comparing air dried would indeed show a difference } but if you really were comparing it to critical point dried samples, could } you elaborate a bit on the nature of those differences? And would you } have any comment about differences in contrast comparing ESEM vs. gold } coated images. } } } Disclaimer: SPI Supplies offers critical point dryers and sputter coaters so } I guess we would be a bit nervous of any technology that should evolve that } would render obsolete at the same time both CPD units and sputter coaters! } } Chuck } ============================================ } } Charles A. Garber, Ph. D. Ph: 1-610-436-5400 } President 1-800-2424-SPI } SPI SUPPLIES FAX: 1-610-436-5755 } PO BOX 656 e-mail:cgarber-at-2spi.com } West Chester, PA 19381-0656 USA } Cust.Service: spi2spi-at-2spi.com } } Look for us! } ######################## } WWW: http://www.2spi.com } ######################## } ============================================ }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
I need to do some high temperature TEM studies and am looking for a suitable mounting resin. I will be embedding and microtoming powder materials and would like to go as high in temperature as possible. The materials themselves can withstand the temperatures but I doubt my conventional epoxy would survive.
Is there any silicone epoxy or other suitable material designed to withstand high temperatures (maybe as high as 600-800C) for a short time for embedding?
Any suggestions you can offer are greatly appreciated.
Thanks in advance!
George R. Munzing Jr. Engelhard Corporation Strategic Technologies Group 25 Middlesex/Essex Turnpike Iselin, NJ 08830 Tel: 732-205-7030 FAX: 732-205-5300
There is another 'glue' that allows you to tailor the ion milling rate. The product is called Araldit or LEIT-C. It is a carbon based glue that starts as a powder, transitions into a viscous liquid, then polymerizes shortly thereafter. The advantage is the glue mills at a slightly lower rate than normal epoxies, and you can increase the carbon content to further reduce the milling rate. South Bay Technology does supply this type of product. Please let me know if you would like further information.
Best Regards, ---------------------------------- Shane Roberts Applications Engineer South Bay Technology,Inc. 'Materials Processing Solutions' www.southbaytech.com phone: 949.492.2600 fax: 949.492.1499 email: roberts-at-southbaytech.com -----------------------------------
-----Original Message----- } From: Isabel Nogueira [mailto:isabeln-at-popsrv.ist.utl.pt] Sent: Monday, October 14, 2002 9:16 AM To: Microscopy-at-sparc5.microscopy.com
Hello,
I need to prepare TEM cross-section samples of alternate SiO2 and TiO2 layers on a Si substrate. For that purpose I would appreciate suggestions on glues, suitable for TEM work, that have a sputtering rate not too different from my non homogeneous layers. The final sputtering will be on an Ion Mill (4keV) using Ar+ ions. All suggestions are welcome (including vendors) either to the list or to me directly. Thank you,
Isabel
Isabel Nogueira Instituto Superior Técnico Dep. Materiais Avenida Rovisco Pais 1049-001 Lisboa Portugal tel.: +351 218418123 fax: +351 218418120 email: isabeln-at-popsrv.ist.utl.pt
Dear All who replied, Using both en block staining for the tissues followed by double staining of grids with UA and Lead citrate seems to be a good idea. Anyway, to get a personal insight and experience, I will try fixation with and without en-block staining. Thanks again. M. Yousuf.
I saw the first post this weekend and am jut now getting to it - doesn't look like the answer is really been quite addressed yet... and yes even at the risk of starting something . . . . ;)
What a wonderful question. Another semantic, logical, debatable but ultimately one answer question!
Compound microscopes are as you say anything with a compound series of lenses. Yes a 'Stereo' microscope should be called a compound stereo microscope (or some variation). The distinction is that (I feel) the "compound" microscope should be call a transmitted (or epi-illuminated) light microscope. But then that would demonstrate an inclination of an electron microscope bias.
There is nothing wrong with the term stereomicroscope, but the issue with changing the compound microscope's name is more difficult than worth it. It is much like trying to get the rest of the US to start using the metric system. The terminology is too entrenched, but it works, so why fix it?
The term binocular then becomes similarly confusing as Stereo microscopes are also 'binocular' the fixed definition isn't much better than the 'compound' in that it has the same issues. But then as addressed before binocular compound 'transmitted' microscopes are not 'stereo' microscopes.
Greg, call 'em what you want - as long as your definition is made clear to your audience and the distinctions made in the discussion on the list server is kept in mind.
Geoff Williams Microscopy Facility Supervisor
} -----Original Message----- } From: by way of Ask-A-Microscopist [mailto:gbarclay-at-trinidad.net] } Sent: Saturday, October 12, 2002 2:48 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Ask-A-Microscopist:LM terminology } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (gbarclay-at-trinidad.net) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, } October 11, 2002 at 23:40:22 } ------------------------------------------------------------------------ -- } - } } Email: gbarclay-at-trinidad.net } Name: Greg Barclay } } Organization: University of the West Indies } } Education: Graduate College } } Location: St. Augustine, Trinidad, West Indies } } Question: Light microscope terminology: We all know what a } stereomicroscope is and what a compound microscope is, but are they } not technically both the same thing? Should not the distinction be } between the "simple microscope," basically a highly corrected doublet } or triplet lens, forming the Dowland type of microscope, and any kind } of microscope with a separate eyepiece and objective lens? If so, } then both the stereo and binocular (or monocular) microscopes are } compond microscopes. I have been referring to "stereomicroscopes and } binocular microscopes" in our Life Sciences Dept, and having some } arguments with those who refer "stereomicroscopes and compound } microscopes," over what is really what. } } ------------------------------------------------------------------------ -- } -
Hello everyone, although this is strictly not a microscopy question, I'm posting this in the hope that someone has been through this process already and can offer some advice.
Our LEO 360FE SEM computer saves images on a Panasonic LF-7300 WORM drive. It doesn't seem feasible to connect the computer to our intranet directly (too old/difficult!) but a relatively straighforward way to get at the images is to install an identical WORM drive on a PC which is networked. However I imagine that some kind of software is needed to make the drive functional under Windows NT. My internet searches for drivers have drawn blanks so far. Has anyone done this and can give me some advice? We are asking LEO for some support on this but independent advice is always very valuable.
======================================================================= This e-mail is intended for the person it is addressed to only. The information contained in it may be confidential and/or protected by law. If you are not the intended recipient of this message, you must not make any use of this information, or copy or show it to any person. Please contact us immediately to tell us that you have received this e-mail, and return the original to us. Any use, forwarding, printing or copying of this message is strictly prohibited.
No part of this message can be considered a request for goods or services. ======================================================================= Any questions about Bookham's E-Mail service should be directed to postmaster-at-bookham.com.
Here is a colleague's method for using aluminum foil on SEM stubs for a smooth background:
1. "Reynolds" seems smoothest and "regular" seems easiest to work with.
2. Burnishing. I've tried various things and they usually end up scratching the foil. To make sure it is absolutely smooth flat, I carefully cut a strip that is a bit wider and longer than a stub. Place it on a clean surface, cover it with something very smooth (weighing paper works well) and work over it with a finger tip to get it absolutely as flat as possible. Picking it up by the very edge with tweezers helps avoid wrinkles.
3. The smooth, flat piece is then transferred to a piece of double-stick scotch tape and once more pressed to the tape to get a very flat surface with no bubbles between foil and tape. This should be done on glass or a surface from which tape plus foil can then be carefully lifted.
4. The tape plus foil can then be applied to the stub - if it still looks nice, proceed to step five. If wrinkles or bumps have appeared, pull it up and try again. (You can put the tape directly on the stub and then apply the foil - However, I have found that it works best for me to have a nicely bonded duo of smooth tape and foil first)
5. Using weighing paper, again, press tape plus foil gently to the stub.
6. Finally, use a single-edge razor blade to trim off the excess foil and tape around the edge of the stub. You can apply a couple of teeny dabs of silver paste at several edge points if you want to make absolutely certain the foil is well connected to the stub. I have gotten into the habit of making the strip of foil plus tape slightly narrower than stub diameter so that there is a small crescent of stub above and below the strip, simply as a way of telling top and bottom at a glance.
7. Additional note. For mounting an object with a convex (like my little larval shells) or irregular lower surface, aluminum foil can be gouged with a minuten insect pin before applying silver paste or paint to help achieve better contact and bonding.
As with everything, it just takes a bit of experimentation. . . .
Greetings Sundeep, If you try entering "polarized fluorescence microscopy" into the Google search engine you will get some hits. Using a polarized light source in conjunction with a crossed polarizing filter which can be rotated (analyzing filter) is a technique used in different approaches to fluorescence microscopy to observe molecular rotations in protein folding or in TIRF applications. It can be used in conjunction with FRET type experiments. I believe the book entitled "Fluorescence Microscopy" published by Bios Scientific may have a short section which discusses polarized fluorescence microscopy. Regards, Karl G.
At 06:11 PM 10/10/2002 -0500, sundeep.bhandari-at-nikon.co.uk wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_______________________________________________ Karl Garsha Light Microscopy Specialist Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illiniois at Urbana-Champaign 405 North Mathews Avenue Urbana, IL 61801 Room B650J Tel: (217) 244-6292 Fax: (217) 244-6219 www.itg.uiuc.edu
We also have an Olympus Model SZH10 that has only one "objective" but it's not really an objective lens [1X to 80X total mag. for microscope]. It's simply a mag. doubler in front of the pair of objective lenses. If one turns the microscope upside down and removes this mag. doubler, they would see two objectives and there are two optical paths all the way up to the eyepieces. Ain't no way to have stereo with one optical path. I've never seen a "metallurgical" "compound" microscope that has mags. up to 1 KX. with two distinct optical paths.
Peter
-----Original Message----- } From: Mike Bode [mailto:mb-at-Soft-Imaging.com] Sent: Monday, October 14, 2002 4:36 PM To: 'Microscopy-at-MSA.Microscopy.Com'
I think, Peter and you are right with the statement that different light paths are necessary to achive a true stereo effect. I am not so sure, that 2 separate objectives are really required. I know of at least one company (I have no affiliation with or interest in that company) that makes a 3D microscope and I think they are using only one objective lens (http://www.edge-3d.com).
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: John W. Raffensperger, Jr. [mailto:johnr-at-helwigcp.com] Sent: Monday, October 14, 2002 8:59 AM To: 'by way of Ask-A-Microscopist'; Microscopy-at-sparc5.microscopy.com
Just because a microscope has two eyepieces, does not mean that it is a "stereo" microscope.
A stereo microscope has not only two eyepieces, but also two objectives, and two separate light paths. Because of having two slightly offset images, you see a three dimensional image. Usually these are lower powered systems, but still compound microscopes (multiple lens systems).
On a high power (relatively) microscope with a binocular head, you only have one objective, and one light path from the specimen to the head. The prism(s) in the head divide the light path to provide multiple images, but they are all (three in the case of an extra camera port) identical. This does not provide a 3-D image, it just makes it easier on the eyes and brain to process the image because one eye is not distracted.
I am not a scientist, and would be interested in any corrections to the above.
John W. Raffensperger, Jr. IS Manager Helwig Carbon Products, Inc.
Question: Light microscope terminology: We all know what a stereomicroscope is and what a compound microscope is, but are they not technically both the same thing?
In the history of microscopy, the term "compound microscope" was used in contrast to a "simple microscope." A simple microscope had one lens, while a compound microscope had two or more lenses (for example, an eyepiece). Thus in the 1600s, Leeuwenhoek used a simple microscope (one lens of rather high magnification), while Hooke did his work with a compound microscope (which had an eyepiece). Compound microscopes were much more convenient to use and were easier on the eyes. On the other hand, simple microscopes were less affected by chromatic abberation, which allowed Leeuwenhoek to see smaller objects (bacterial, sperm, and so forth) than could be seen with the compound microscopes of the time. Not until the 1820s, with the advent of achromatic lenses for compound microscopes, could microscopists improve much on the detail of Leeuwenhoek's observations.
Kent
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ A. Kent Christensen, Professor Emeritus Department of Cell and Developmental Biology, Medical Science II Building University of Michigan Medical School, Ann Arbor, MI 48109-0616 Tel (work) (734) 763-1287, Fax (work) (734) 763-1166 akc-at-umich.edu http://www.umich.edu/~akc/ ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
--On Tuesday, October 15, 2002 4:14 PM -0400 Geoff Williams {willi1gl-at-cmich.edu} wrote:
} I saw the first post this weekend and am jut now getting to it - doesn't } look like the answer is really been quite addressed yet... and yes even } at the risk of starting something . . . . ;) } } What a wonderful question. Another semantic, logical, debatable but } ultimately one answer question! } } Compound microscopes are as you say anything with a compound series of } lenses. Yes a 'Stereo' microscope should be called a compound stereo } microscope (or some variation). The distinction is that (I feel) the } "compound" microscope should be call a transmitted (or epi-illuminated) } light microscope. But then that would demonstrate an inclination of an } electron microscope bias. } } There is nothing wrong with the term stereomicroscope, but the issue } with changing the compound microscope's name is more difficult than } worth it. It is much like trying to get the rest of the US to start } using the metric system. The terminology is too entrenched, but it } works, so why fix it? } } The term binocular then becomes similarly confusing as Stereo } microscopes are also 'binocular' the fixed definition isn't much better } than the 'compound' in that it has the same issues. But then as } addressed before binocular compound 'transmitted' microscopes are not } 'stereo' microscopes. } } Greg, call 'em what you want - as long as your definition is made clear } to your audience and the distinctions made in the discussion on the list } server is kept in mind. } } Geoff Williams } Microscopy Facility Supervisor } } } } -----Original Message----- } } From: by way of Ask-A-Microscopist [mailto:gbarclay-at-trinidad.net] } } Sent: Saturday, October 12, 2002 2:48 PM } } To: Microscopy-at-sparc5.microscopy.com } } Subject: Ask-A-Microscopist:LM terminology } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } } submitted by (gbarclay-at-trinidad.net) from } } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, } } October 11, 2002 at 23:40:22 } } } ------------------------------------------------------------------------ } -- } } - } } } } Email: gbarclay-at-trinidad.net } } Name: Greg Barclay } } } } Organization: University of the West Indies } } } } Education: Graduate College } } } } Location: St. Augustine, Trinidad, West Indies } } } } Question: Light microscope terminology: We all know what a } } stereomicroscope is and what a compound microscope is, but are they } } not technically both the same thing? Should not the distinction be } } between the "simple microscope," basically a highly corrected doublet } } or triplet lens, forming the Dowland type of microscope, and any kind } } of microscope with a separate eyepiece and objective lens? If so, } } then both the stereo and binocular (or monocular) microscopes are } } compond microscopes. I have been referring to "stereomicroscopes and } } binocular microscopes" in our Life Sciences Dept, and having some } } arguments with those who refer "stereomicroscopes and compound } } microscopes," over what is really what. } } } } } ------------------------------------------------------------------------ } -- } } - }
Does anyone have a recipe for mouse brain perfusion and fixation? I am trying to get the best brain preservation in mice. I trying to prevent neurons from swelling and I need to look at the Golgi apparatus. How about calcium chloride as an additive?
Thanks!
Karen
Karen L. Bentley, M.S.(previously Jensen) Associate Scientist & Project Manager Electron Microscope Research Core University of Rochester Medical Center Rochester, NY 14642 585-275-1954
If your drive is the old WORM technology, good luck finding one! I haven't seen one for sale in years.
What computer does the 360 use? I have forgotten... Even if it is a DEC (perhaps especially) it would be easier to send the data to a newer PC for storage on hard drives and CD-Rs.
Regards, Woody
Woody White McDermott Technology Inc. McD: http://www.mtiresearch.com/ Mine: http://woody.white.home.att.net
-----Original Message----- } From: Richard Beanland [mailto:richard.beanland-at-bookham.com] Sent: Wednesday, October 16, 2002 9:12 AM To: 'Microscopy-at-MSA.Microscopy.Com'
Hello everyone, although this is strictly not a microscopy question, I'm posting this in the hope that someone has been through this process already and can offer some advice.
Our LEO 360FE SEM computer saves images on a Panasonic LF-7300 WORM drive. It doesn't seem feasible to connect the computer to our intranet directly (too old/difficult!) but a relatively straighforward way to get at the images is to install an identical WORM drive on a PC which is networked. However I imagine that some kind of software is needed to make the drive functional under Windows NT. My internet searches for drivers have drawn blanks so far. Has anyone done this and can give me some advice? We are asking LEO for some support on this but independent advice is always very valuable.
======================================================================= This e-mail is intended for the person it is addressed to only. The information contained in it may be confidential and/or protected by law. If you are not the intended recipient of this message, you must not make any use of this information, or copy or show it to any person. Please contact us immediately to tell us that you have received this e-mail, and return the original to us. Any use, forwarding, printing or copying of this message is strictly prohibited.
No part of this message can be considered a request for goods or services. ======================================================================= Any questions about Bookham's E-Mail service should be directed to postmaster-at-bookham.com.
from my experience with mouse (CNS; spinal chord, muscle, testis and other tissue) I would strongly recommend a mixture of 2% paraformaldehyde / 4% glutaraldehyde (made from "the good one" prepared after"Anderson")and a short (!) pre-perfuse (as you name it) to flush out all blood cells, which otherwise regularly clog and occlude the fine capillaries. However, though simple saline flush can be sufficient, I routinely use a mixture of Heparin (anti-coagulant), Procain (keeps the blood vessels "open"), PVP (polyvinylpyrrolidon, MW-class 30 K; for osmotic pressure) in aqua dest. and pH-adjustment. Method published originally in a wonderful perfusion-technique paper by Forssmann, WG et al (1977) in Anat. Rec. 188: p 307- 314. Just follow their technical procedure exactly (except that in the small mouse you have to use a 26 gauge butterfly teflon-needle) and you`ll get excellent fixation. good luck! peter ********************************** peter.heimann-at-uni-bielefeld.de Dr. Peter Heimann Developmental Biology & Molecular Pathology; W7-107 University of Bielefeld D 33501 Bielefeld / Germany phone: xx49(0)521-106-5628 / 5627 FAX : " " - 5654 www.uni-bielefeld.de/biologie/Entwicklungsbiologie/ www.uni-bielefeld.de/SFB549 ***********************************
I perfuse mouse with 1x PBS (20 mM Na-phosphate, 150 mM NaCl, PH 7.4) 15 ml. 2 ml/min, room temp, then same amount/speed 0.1% GA + 2% formaldhyde in 1x PBS, room temp. Brain as quick as possible removed from the scull on ice and sliced 0.5 mm thick. Slices individually fixed in 1% GA/2% formaldehyde/1x PBS at +4oC overnight.
I did not check Golgi but mitochondria and other stuff is OK. Sergey
P.S. I did not try Ca+2, but it's good combination with formaldehyde for brain.
P.P.S. All chemicals are EM grade and fixers prepared just before use. Slicing is critical. Speed of preparation is VERY critical. The way you kill animal is also critical. Godd luck!
At 03:56 PM 10/16/02 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Just passing this along for Matt who is not on the listserver. Please email him directly with any ideas.
Many Thanks!
Al Coritz Electron Microscopy Sciences
I was wondering if you knew any good protocols for decalcifying the mouse skull. We haven't done too much of it, but when we have, we've used 3%HCl and 15%NaCl. As always, we're looking for a way to keep shrinkage of the brain to a minimum. Do you have any ideas? Any help would be appreciated. Thanks a lot. Matt
Matt McElwee Research Specialist Department of Surgery K4/617 Clinical Science Center 600 Highland Avenue Madison, WI 53792 mcelwee-at-surgery.wisc.edu
Karen, Shrinkage or swelling of course is related to the osmolarity of the fix (and vehicle). I like to run a variety of fixes ie 2.5% GA vs 2% PF + 1.25% GA, etc. I think a good buffer would be 0.1 M Hepes and definitely add 3 mM CaCl2 for membrane tonicity. When osmicating it is important to use KFECN6 (0.8%) reduced osmium (1%), this keep the myelin sheaths from blebing (on ice). I also keep the CaCl2 in the osmium step. I also like long en-bloc staining (2% filtered UA for 2 hrs). This gives good contrast and helps stabilize membranes during dehydration. Remember even though you perfused to cut small pieces (3 mm cubed) for infiltration of subsequnent solutions. If you want to see actin or neurofilaments use 0.15% tannic acid after the osmium (about 5 min only)-Kent McDonald- With tissue I like to go 1:1 Propylene Oxide to Epon overnight. Good Luck.
Mike Delannoy JHMI Microscopy Facility
"Jensen, Karen" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Listers: } } Does anyone have a recipe for mouse brain perfusion and fixation? I am } trying to get the best brain preservation in mice. I trying to prevent } neurons from swelling and I need to look at the Golgi apparatus. How about } calcium chloride as an additive? } } Thanks! } } Karen } } Karen L. Bentley, M.S.(previously Jensen) } Associate Scientist & Project Manager } Electron Microscope Research Core } University of Rochester Medical Center } Rochester, NY 14642 } 585-275-1954
You might also check this page http://sourceforge.net/
Pavel
----- Original Message ----- } From: "Pijpers,Frank" {fpijpers-at-nl.feico.com} To: "Microscopy-at-MSA.Microscopy.Com" {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, October 17, 2002 9:04 AM
we have an agfa duoscan T2500. we would like to better automate our scanning. does anyone know of a method of auto scaning negs on the duoscan. thank s john
we have an agfa duoscan T2500. we would like to better automate our scanning. does anyone know of a method of auto scaning negs on the duoscan. thank s john
We are going to move our EM lab to a new place (in the same building) and are planning to put an anti-vibration platform under the column of our FEG-ESEM XL30. Do we have to place a HV transformer on the same platform (the transformer is right behind the column)?
Any additional advise will be very helpful also, especially experience with anti-vibration platforms of different manufacturers.
Right now I can routinely make micrographs at magnifications only up to 50,000. After hours, when vibrations are lower, I can (sometimes) go up to 100,000-150,000. Can platform improve performance of microscope in this range of magnifications?
Thank you,
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} } } FotoLook has to be run as an independent application (not from within } Photoshop). } Batches can be defined here. } To repeat the same batch, drag the contents of the Done folder to the ToDO } folder and run the batch again. } } At 11:22 AM 10/17/2002 -0400, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/
Hello all - anybody have spare parts for the Technics MIM-IV ion mill, or know where I can find them? The anode tips in our ion guns (TSA 1-aperture) are pretty hopelessly worn out. Any help would be greatly appreciated!
Rick
{} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} Rick Hugo, Ph.D. Research Associate, Geomicrobiology and Electron Microscopy Lab Department of Geology, Portland State University Portland, OR 97207-0751 {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {}
At 07:50 AM 10/17/02 -0400, Everett Ramer {eramer-at-cellomics.com} wrote: } I am looking for a collection of highly optimized source code (in C) for the } basic image processing functions.
Optimized for what?
**Please note new department name**
Kevin Frischmann, Laboratory Manager Microscopy & Imaging Facility American Museum of Natural History Central Park West at 79th Street New York, NY 10024-5192 USA
UA is soluble in veranol acetate buffer at pH 4.5 and below.
However, UA is INSOLUBLE in cacodylate buffer at 4.5 and 3.5.
UA is soluble in cacodylate buffer at pH 0 and 0.5.
I don't know what happens between 0.5 and 3.5, but who wants to use a buffer at that pH anyway? Somebody may have publised this info, but I really don't need to know enough to chase it down. If one wanted to stain at that low pH, cacodylate doesn't have much buffering capacity down there.
Don't quote me on the exact pH's. I just tried this with pH paper and tiny drops, not a pH meter.
I still recommend washing out the caco with the buffer that the UA is in (or water) before using adding UA---that is if you choose to do en bloc staining.
Sara Miller
On Tue, 15 Oct 2002, gang ning wrote:
} One thing I want to point out is people usually don't use UA in } cacodylate buffer is NOT for the reason that UA is NOT SOLUBLE in it, } but for the reason that UA works better in buffers of low pH. } } saram-at-duke.edu wrote: } } } Again, see CAPS below. } } (CAPS ARE USED TO DIFFERENTIATE COMMENTS, NOT TO CONVEY HOSTILITY.) } } } } } } On Sun, 13 Oct 2002, gang ning wrote: } } } } } Thanks for the clear definition of biohazard. But it is obvious that } } } 1) UA is a nasty/hazard chemical. Everyone wants to avoid it if possible, } } } } } } } UA CERTAINLY CAN BE CALLED NASTY. IT DOES REQUIRE SPECIAL HANDLING } } PROCEDURES, AS DO OTHER CHEMICALS THAT WE USE AND CAN BE HANDLED SAFELY } } UNDER THE PROPER CONDITIONS. NOT EVERYONE WANTS TO AVOID IT. FOLKS MUST } } MAKE THEIR OWN JUDGEMENT AS TO WHETHER THE EXTRA PROCESSING TIME AND CARE } } OF HANDLING ANOTHER CHEMICAL IS WORTH IT TO THEM. NOT USING UA IS FINE. } } HOWEVER, DO NOT MAKE A GENERALIZATION ABOUT *EVERYONE* JUST BECAUSE YOU } } CHOOSE NOT TO USE IT. } } } } } 2) it does take at least 1 hr + time for washing thoroughly (how long?) } } } for a 1 mm3 block .... it is clearly time-consuming procedure } } } } } } } IT DOES REQUIRE EXTRA TIME (1-2 HOURS). ONE CAN STAIN A 1 mm3 BLOCK FOR } } 30-60 MIN AND WASH FOR 30-60 MIN. } } } } unless your turn around time is not an issue and working overtime is not a } } } } } matter with your hospital policy, } } } } } } } TIME IS VERY MUCH AN ISSUE FOR US IN THE HOSPITAL SETTING. OUR TURN } } AROUND TIME IS *ROUTINELY* NEXT DAY FOR SAMPLES FIXED IN GLUT, OS, UA. } } TISSUE COMES IN IN THE AM, IS PROCESSED THAT DAY, BAKED OVERNIGHT, CUT AND } } VIEWED THE NEXT DAY, AND PATHOLOGISTS HAVE MICROGRAPHS *IN HAND* THAT } } AFTERNOON--ALMOST ALWAYS. MY TECHS RARELY HAVE TO WORK OVERTIME, AND WHEN } } THEY DO, IT'S USUALLY BECAUSE OF EXTRA CUTTING OF HAVING TO RETRIEVE } } SPECIMENS OUT OF PARAFFIN BLOCKS OR OFF SLIDES, NOT BECAUSE THEY PUT } } SOMETHING INTO UA. } } } } YOU ARE CLEARLY VERY NEGATIVE TOWARD THE USE OF UA. FINE. DON'T USE IT. } } BUT DON'T BE SARCASTIC ABOUT SPECIMEN TURN AROUND TIME AND } } TECHNICIAN OVERTIME, BECAUSE IT JUST ISN'T AN ISSUE IF SPECIMENS AND } } WORKLOADS ARE MANAGED PROPERLY. } } } } } 3) en block staining has adversary effect on showing glycogen so is not } } } suitable for muscle, heart, liver and so on; there is not significant } } } meaning for kidney .. then what else left? LOTS: } } } } } } } BRAIN, CILIA, TUMORS, VIRUSES, MUSCLE DISEASES OTHER THAN GLYCOGEN STORAGE } } DISEASES, RESEARCH SAMPLES, ETC. } } } } } } It is overkill for solid tissue blocks, IT IS NOT OVERKILL WHEN THE } } BEST PRESERVATION, RATHER THAN DECREASING PROCESSING TIME IS OF } } IMPORTANCE. } } } } and 4) ethanolic/methanolic UA of higher concentration penetrates tissue } } } } } much faster than other aqueous UA solutions and gives fresher image and } } } better contrast but sometimes leave precipitation around cell member no } } } matter how one washes before staining. } } } } } } } AS I SAID BEFORE, IF THE CORRECT BUFFER IS USED SO THAT UA DOES NOT } } PRECIPTATE WITH IT, THERE IS NO PRECIPITATION. PERHAPS YOUR *HIGHER } } CONCENTRATION* IS A CAUSE OF DIFFICULTY IN WASHING. SINCE UA IS SOLUBLE } } IN ALCOHOL, UNBOUND, UNPRECIPITATED UA IS EASILY WASHED OUT BY THE } } DEHYDRATING AGENT(S). IF YOU HAVE PRECIPITATE, IT IS MOST LIKELY DUE TO } } SOME REACTION WITH THE SOLUTION LEFT IN THE CELL BEFORE YOU ADD THE UA, OR } } PERHAPS THE *HIGHER* CONCENTRATION YOU USE ISN'T WASHED OUT??? } } } } } For EM work, the simpler and better, as long as it works. } } } } } } } YES, THE KEY IS, *AS LONG AS IT WORKS* FOR THE PURPOSE YOU HAVE. IF THE } } PURPOSE IS TO PRODUCE THE BEST ULTRASTRUCTURE, AND IF MEMBRANES AND OR } } LIPIDS ARE PART OF THE QUESTION, USE OF UA IS GOOD. } } } } Overkill is always a problem. YES, NO ONE WANTS TO SPEND MORE TIME THAN } } IS NECESSARY OR USE CHEMICALS THAT CAN BE HAZARDOUS IF NOT HANDLED } } PROPERLY, BUT UA IS NOT ALWAYS OVERKILL. } } } } IT DEPENDS ON YOUR APPLICATION. EACH TO HIS OWN. } } } } } } } } } } } saram-at-duke.edu {mailto:saram-at-duke.edu} wrote: } } } } } } } See CAPS interspersed below for distinction from the original. } } } } } } } } Sara Miller } } } } } } } } On Fri, 11 Oct 2002, Gang Ning wrote: } } } } } } } } } ------------------------------------------------------------------------ } } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com {mailto:ListServer-at-MSA.Microscopy.Com} } } } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } -----------------------------------------------------------------------. } } } } } } } } } } } } } } } Hi } } } } } } } } } } I agree with Dr. Sara Miller's point that UA en block staining in some extent } } } } } helps in preservation of ultrastructure as well as enhancement of the } } } } } contrast. However, it also has disadvantages such as one has to deal with UA, } } } } } a biohazard reagent, } } } } } } } } } A BIOHAZARD IS A BIOLOGICAL AGENT THAT IS HAZARDOUS (INFECTIOUS). UA IS } } } } RADIOACTIVE AND IS A HEAVY METAL. IT, LIKE GLUTARALDEHYDE, OSMIUM, LEAD, } } } } RESINS, PROPYLENE OXIDE, ETC., SHOULD BE HANDLED WITH CARE. HANDLED } } } } PROPERLY, IT IS NOT ANY MORE DANGEROUS THAN THE OTHER NASTY CHEMICALS } } } } ELECTRON MICROSCOPISTS HAVE TO USE. IT IS NOT A BIOLOGICAL AGENT. } } } } } } } } and sometimes background from crystal precipitation, } } } } } } } } } especially when ethanolic UA is used. } } } } } } } } } WE NEVER SEE PRECIPITATION. PERHAPS TRY DILUTING THE STAIN OR WASHING } } } } THOROUGHLY BEFORE USING IT WILL ELIMINATE THE PRECIPITATE. UA IS NOT } } } } SOLUBLE IN PHOSPHATE OR CACODYLATE BUFFER; IF YOU DON'T WASH OUT THESE } } } } BUFFERS BEFORE ADDING UA IN VERANAL ACETATE BUFFER, UA IN WATER, OR UA IN } } } } ETHANOL OR METHANOL, YOU CAN GET PRECIPATE. } } } } } } } } More importantly, it is time-consuming } } } } } } } } } because it takes hours for aqueous UA solution to penetrate a sizable tissue } } } } } block. } } } } } } } } } WE USE 1 MM TISSUE BLOCKS AND STAIN FOR AN HOUR 1% UA IN 0.11M VERONAL } } } } ACETATE BUFFER. IF THE TISSUE OR CELL PELLET IS SMALLER, YOU CAN GET AWAY } } } } WITH 30 MIN IN UA. } } } } } } } } Therefore, I would say it all depends your tissue and purpose of your } } } } } } } } } analysis. } } } } } } } } } I AGREE. IT ALL DEPENDS ON YOUR PURPOSE. IF YOU WANT BEAUTIFUL } } } } ULTRASTRUCTURE AND YOU'RE NOT LOOKING SPECIFICALLY FOR GLYCOGEN, USE UA EN } } } } BLOCK. IF YOU'RE IN A HURRY OR YOU CARE TO SEE ALL THE GLYGOGEN, DON'T } } } } USE IT. } } } } } } } } } In my lab, we routinely do en block with samples of blood, tissue culture, } } } } } virus, bacteria and other microorganisms, which are believed more vulnerable } } } } } to solvent, but not with solid tissue. We have a lot of samples from kidney } } } } } biopsy and we didn't find en block made any difference from staining thin } } } } } sections whereas taking your time to have a sufficient osmication is much } } } } } more helpful to your structure. } } } } } } } } } } Greg Ning } } } } } } } } } } } } } } } } } } } } mohammed y abdulrawoof / f40z006 75760 wrote: } } } } } } } } } } } ------------------------------------------------------------------------ } } } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com {mailto:ListServer-at-MSA.Microscopy.Com} } } } } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } -----------------------------------------------------------------------. } } } } } } } } } } } } Dear listers, } } } } } } Please advise as to the usefulness/advantage of doing enblock staining of } } } } } } biological tissues as against doing heavy-metal staining of grids with } } } } } } thin sections. The tissues that I am about to process are for diagnosis } } } } } } (pathology). } } } } } } } } } } } } Thanks } } } } } } M. Yousuf Abdul-Rawoof } } } } } } } } } } } -- } } } } } Gang Ning, M.D., Ph.D. } } } } } Director } } } } } Electron Microscopy Facility } } } } } Medical College of Wisconsin } } } } } 8701 Watertown Plank Road } } } } } Milwaukee, Wisconsin 53226 } } } } } (414) 456-8344 } } } } } e-mail: gning-at-mcw.edu {mailto:gning-at-mcw.edu} } } } } } http://www.mcw.edu/cellbio/emfacility.html } } } } } } } } } } } } } } } } } } } } } } } } } } } } } Sara E. Miller, Ph. D. } } } } P. O. Box 3712 } } } } Duke University Medical Center } } } } Durham, NC 27710 } } } } Ph: 919 684-3452 } } } } FAX: 919 684-3265 } } } } } } } } } } } -- } } } } } } Gang Ning, M.D., Ph.D. } } } } } } Director } } } } } } Electron Microscopy Facility } } } } } } Medical College of Wisconsin } } } } } } 8701 Watertown Plank Road } } } } } } Milwaukee, WI 53226 } } } } } } (414) 456-8141 } } } } } } } } } } } } } } } } Sara E. Miller, Ph. D. } } P. O. Box 3712 } } Duke University Medical Center } } Durham, NC 27710 } } Ph: 919 684-3452 } } FAX: 919 684-3265 } } } } } } -- } } Gang Ning, M.D., Ph.D. } } Director } } Electron Microscopy Facility } } Medical College of Wisconsin } } 8701 Watertown Plank Road } } Milwaukee, WI 53226 } } (414) 456-8141 } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
I have put together an extensive collection of links to all kinds of microscopy and imaging resources on the WWW and there are several links to software and source code repositories:
============================================== Everett Ramer wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I am looking for a collection of highly optimized source code (in C) for the } basic image processing functions. } } Everett Ramer } Cellomics, Inc. } Pittsburgh, PA
13.45-14.30 "Creating, Viewing and Storing Large Image Data Fields" by Alan Entwistle, (Ludwig Institute, University of London)
14.30-14.45 "Development of an Image Analysis Program for Measuring Bone Resorption in vitro" by Rob van't Hof and Miep Helfrich (University of Aberdeen Medical School)
14.45-15.00 "Microscopy of Tick Salivary Glands" Barry Stewart (Zoology, University of Aberdeen)
15.00-15.30 TEA
15.30-16.15 "Microscopy and variant Creutzfeldt-Jakob (vCJD) disease" by James Ironside, (CJD Surveillance Unit, Edinburgh)
16.15-16.30 'Iso-butanol/water: a new staining technique for light and electron microscopy by Ian Roberts', (Scottish Crop Research Institute, Dundee)
16.30 Presentation of Prizes
Electron Microscope unit School of Biological Sciences University of Aberdeen Aberdeen AB24 2TZ
Tel 01224-272847 Fax 01224-272396 ------------ Kevin Mackenzie k.s.mackenzie-at-abdn.ac.uk
I have the task of preparing TEM foils from 1 mm. O.D. 18K and 14K Gold wire. I was successful in ion milling dimpled 50um section/foils in PIPS at 4Kv., 4 degree angle. We are looking at the strain fields about 100nm. precipitates. The ion milling caused damaged to the structure as indicated by the ripple patterns in the material as viewed in the TEM. Obviously this may/can alter our examination.
Next, I would like to try electropolishing the material if I can make suitable foils for the polisher. We cold rolled the wire, and punched out 3 mm. disks to work with. Electropolishing is the next step to try.
What I am asking, does anyone know of an electropolishing solution for Au, suitable for a Tenupol 5 or a Fischione machine?
Thanks in advance
Fred Pearson
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
Listers, I'm forwarding this answer to the list because of our recent discussion on UA and the question posed by Phil about UA inactivation of prions.
The question at a recent microscopy meeting concerned whether osmium "killed" (inactivated) prions. It arose because of a question about how specimens suspected of containing prion material (Creutzfeld-Jacob Disease and other spongiform encephalapathies) should be handled in the EM lab.
The discussion centered around osmium inactivation because it is a proven fact that aldehydes DO NOT inactivate prions. Osmium is usually the next step, and most EM folks use it; some do not use UA. The osmium inactivation question was also visited on this net some years ago.
My answer was that some oxidizing agents (peracetic acid; Vet J 159:10, 2000) reduce activity of prions in brain, and thus, one might suspect that osmium could act similarly. This article does say that many oxidizing agents tried have no effect, and that the effect is related to whether the prions are aggretated (the more aggregated, as in homogenized specimens, the less inactivated). Peracetic acid works better on brain than on homogenized tissue. However, I pointed out that in a personal communication with both Drs. D. M. Taylor and Stanley Prusiner, these experts simply said that the experiment of osmium inactivation hasn't been done. We simply don't know. Also, they pointed out that we don't even know whether binding up the prions in resin inactivates them. They recommended working with gloves and collecting the "chips" from trimming for incineration.
Phil has raised the question of whether UA would inactivate prions. His reasoning has merrit. However, I would maintain that, like osmium, UA has not been tested, and that some things that one would think would inactivate prions don't (see ref above). Just because they are stained or bound up doesn't necessarily mean they are inactive.
Radioactivity, even very strong doses, does not inactivate prions. As a matter of fact, this is one reason why folks feel that prions don't contain nucleic acids.
If histology labs have to handle brain suspected of containing prions, they inactivate the tissue with 97% formic acid after formaldehyde fixation (Neurology 40:887, 1990), which doesn't seem too detrimental to structure.
Sara Miller
On Fri, 18 Oct 2002, Philip Oshel wrote:
} Sara, } } I've been mulling over your "does OsO4 oxidize prions?" question that } you mentioned in your talk at the meeting here in Madison this week. } Why OsO4? It's a poor protein stain, so although it's a strong } oxidizer, it wouldn't occur to me to suspect it of degrading prions. } Why not uranyl acetate or (better?) uranyl nitrate? These are good } protein stains, and so should bind better to prions. Further, } although their radioactivity is low, it is present, and as alpha } particles. Nice and heavy, and they do lots of damage at short range. } I should think UAc or UNO3 would be useful for breaking down prions. } (I refuse to say "kill", as I don't think prions have any of the } attributes of life.) } } Mind, I'm not going to do the experiment either. 8-) } } Nice talk! } } Phil } -- } Philip Oshel } Supervisor, BBPIC microscopy facility } Department of Animal Sciences } University of Wisconsin } 1675 Observatory Drive } Madison, WI 53706 - 1284 } voice: (608) 263-4162 } fax: (608) 262-5157 (dept. fax) } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
I need to hear back from those who have said they have software for tracking cells and particle movement. Please let me know if you are ready to bring in demo version.
Regards,
Joe Goodhouse Confocal / EM Core Laboratory Department of Molecular Biology Princeton University 609-258-5432
Visit us at http://www.molbio.princeton.edu/facility/confocal/index.html
Win NT directly supports MO drives. I just went and pulled my old LF- 7303 off the shelf, checked the SCSI ID wasn't in conflict, plugged it into a "Newly rebuilt" NT box, rebooted, and Ta-Da there it was. The SCSI interface itself handles everything.
} If your drive is the old WORM technology, good luck finding one! I haven't seen } one for sale in years. } } What computer does the 360 use? I have forgotten... Even if it is a DEC } (perhaps especially) it would be easier to send the data to a newer PC for } storage on hard drives and CD-Rs. } } Regards, Woody } } Woody White } McDermott Technology Inc. } McD: http://www.mtiresearch.com/ } Mine: http://woody.white.home.att.net } } } -----Original Message----- } } From: Richard Beanland [mailto:richard.beanland-at-bookham.com] } Sent: Wednesday, October 16, 2002 9:12 AM } To: 'Microscopy-at-MSA.Microscopy.Com' } Subject: WORM drives } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello everyone, } although this is strictly not a microscopy question, I'm posting } this in the hope that someone has been through this process already and can } offer some advice. } } Our LEO 360FE SEM computer saves images on a Panasonic LF-7300 WORM drive. } It doesn't seem feasible to connect the computer to our intranet directly } (too old/difficult!) but a relatively straighforward way to get at the } images is to install an identical WORM drive on a PC which is networked. } However I imagine that some kind of software is needed to make the drive } functional under Windows NT. My internet searches for drivers have drawn } blanks so far. Has anyone done this and can give me some advice? } We are asking LEO for some support on this but independent advice is } always very valuable. } } Many thanks, } } Richard } } } _______________________________ } Richard Beanland } Analytical Services } Bookham Technology plc } Caswell, } Towcester, } Northamptonshire NN12 8EQ } UK } Tel: +44 (0) 1327 356362 } Fax: +44 (0) 1327 356398 } http://www.bookham.com } } } } ======================================================================= } This e-mail is intended for the person it is addressed to only. The } information contained in it may be confidential and/or protected by } law. If you are not the intended recipient of this message, you must } not make any use of this information, or copy or show it to any } person. Please contact us immediately to tell us that you have } received this e-mail, and return the original to us. Any use, } forwarding, printing or copying of this message is strictly prohibited. } } No part of this message can be considered a request for goods or } services. } ======================================================================= } Any questions about Bookham's E-Mail service should be directed to } postmaster-at-bookham.com. } } }
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 350 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
Our analytical chemist stated that aluminum foil has a coating of a lubricant on it to facilitate separation from the roll. I was wondering if this has caused anyone any issues with EDX.
Peter Tomic Anadigics, Inc.
-----Original Message----- } From: Tina Carvalho [mailto:tina-at-pbrc.hawaii.edu] Sent: Wednesday, October 16, 2002 1:54 PM To: Microscopy Listserver
Hi, all-
Here is a colleague's method for using aluminum foil on SEM stubs for a smooth background:
1. "Reynolds" seems smoothest and "regular" seems easiest to work with.
2. Burnishing. I've tried various things and they usually end up scratching the foil. To make sure it is absolutely smooth flat, I carefully cut a strip that is a bit wider and longer than a stub. Place it on a clean surface, cover it with something very smooth (weighing paper works well) and work over it with a finger tip to get it absolutely as flat as possible. Picking it up by the very edge with tweezers helps avoid wrinkles.
3. The smooth, flat piece is then transferred to a piece of double-stick scotch tape and once more pressed to the tape to get a very flat surface with no bubbles between foil and tape. This should be done on glass or a surface from which tape plus foil can then be carefully lifted.
4. The tape plus foil can then be applied to the stub - if it still looks nice, proceed to step five. If wrinkles or bumps have appeared, pull it up and try again. (You can put the tape directly on the stub and then apply the foil - However, I have found that it works best for me to have a nicely bonded duo of smooth tape and foil first)
5. Using weighing paper, again, press tape plus foil gently to the stub.
6. Finally, use a single-edge razor blade to trim off the excess foil and tape around the edge of the stub. You can apply a couple of teeny dabs of silver paste at several edge points if you want to make absolutely certain the foil is well connected to the stub. I have gotten into the habit of making the strip of foil plus tape slightly narrower than stub diameter so that there is a small crescent of stub above and below the strip, simply as a way of telling top and bottom at a glance.
7. Additional note. For mounting an object with a convex (like my little larval shells) or irregular lower surface, aluminum foil can be gouged with a minuten insect pin before applying silver paste or paint to help achieve better contact and bonding.
As with everything, it just takes a bit of experimentation. . . .
Hi All: We are looking for a highly qualified, skilled individual to service several TEM's, SEM's and other related electronic equipment across our campus. The position is full time and permanent. The qualified individual should be able to troubleshoot and repair many brands of electronic equipment down to the component level. If interested, please forward a resume to me. Thanks, Michael Coviello Lab Manager, The University of Texas at Arlington, Arlington, TX
I have 4 books ready for review. If you would like to review any of these titles and write your review for the journal please let me know ASAP. Reviewers will be chosen on a first come first serve basis based on your interests. I would also like to know if you would review other books as they come available if your selection has already been sent out.
List:
1) Electron Backscatter Diffraction in Materials Science, Edited by Adam J. Schwartz, Mukul Kumar, and Brent L. Adams
2) Scanning Tunneling Microscopy and its Applications, By C.Bai
3) PCR/RT-PCR in situ, light and electron microscopy, By Gerard Morel and Mireille Raccurt
4) Quick Photoshop for research, a guide to digital imaging for photoshop 4x,5x,6x,7x, by Jerry Sedgewick
If interested please e-mail me along with a mailing address for you. Thank you very much! JoAn
JoAn Hudson, PhD. Institute of Neuroscience 222 Huestis Hall University of Oregon, Eugene, OR 97403
I will apologize in advance in that my recipe is for our Model 550D Single Vertical Jet ElectroPolisher rather than the other systems you mentioned. Seems like there is always a solution with one of our products! ;-)
In any case, I do have a paper from Bernie Kestel titled:
"Jet Thinning of YBa2Cu3Ox High Tc Superconductor and Gold for TEM with a Non-Acid Electrolyte"
This is Technical Report #14 on our online list. Unfortunately, it is not in electronic format, but the reference is Ultramicroscopy 25 (1988) 351-354. I can also send you a photocopy of the paper if that would be easier for you.
While some of our recipes appear to be interchangeable with the twin jet systems. I believe that his BK-2 solution (non-acid electrolyte) is too viscous to be pumped through the twin jet units. However, it may give you a good starting point.
DISCLAIMER: South Bay Technology, Inc. produces equipment and supplies as described above and, therefore, has a vested interest in promoting their use.
The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html
I have the task of preparing TEM foils from 1 mm. O.D. 18K and 14K Gold wire. I was successful in ion milling dimpled 50um section/foils in PIPS at 4Kv., 4 degree angle. We are looking at the strain fields about 100nm. precipitates. The ion milling caused damaged to the structure as indicated by the ripple patterns in the material as viewed in the TEM. Obviously
this may/can alter our examination.
Next, I would like to try electropolishing the material if I can make suitable foils for the polisher. We cold rolled the wire, and punched out 3 mm. disks to work with. Electropolishing is the next step to try.
What I am asking, does anyone know of an electropolishing solution for Au, suitable for a Tenupol 5 or a Fischione machine?
Thanks in advance
Fred Pearson
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 USA
TEL: +1-949-492-2600 Toll-free in the USA: +1-800-728-2233 FAX: +1-949-492-1499
email: henriks-at-southbaytech.com
Celebrating 38 years of providing Materials Processing Solutions for Metallogaphy, Crystallography and Electron Microscopy.
Please visit us online at www.southbaytech.com.
The information contained in this message and any attachments is privileged and confidential. This message is intended for the individual or entity addressed. If you are not the intended recipient, please do not read, copy or disclose this communication. Notify the sender of the mistake by calling +1-949-492-2600 and delete this message from your system.
Thanks to eveyone who vetured to reply to my only half tongue in cheek posting about which microscopes are really compound. I learned that some stereomicroscopes have only one objective lens (!?) which must call some interesting optical designs.
I have received an used Leitz 1512 microtome a few days ago and are not very experienced with it. Everything works perfectly but I can not manage it to set the THICKNESS to MORE THAN 11 my. I can rotate the knob only from 0-11. Is the microtome broken? I do not have a manual for the microtome.
This depends a great deal on the nature and size of the plant tissues. We do a lot of small, herbaceous specimens here, mostly Arabidopsis, but I've done other plant tissues as well, such as roots. The trick is to ignore the directions in the Tousimis manual -- for any biological specimens. What you need to do instead is to let the specimens soak in the liquid CO2 for a given amount of time after first purging the chamber of the absEtOH that was in it when the samples were loaded. Then purge the chamber to replace the lq CO2 with fresh lq CO2, soak, and repeat N times. This is analogous to the dehydration process where the water in the tissues is replaced with EtOH. Now, the EtOH in the tissues is being replaced with lq CO2. Once that is done, then and only then can the samples be dried. Otherwise, they are being dried from EtOH. The length of the soaks and number of soaks is rather empirical and highly dependent on tissue type and size. For the small (a few mm), herbaceous Arabidopsis specimens, we generally use 4 soaks for 10 minutes each. For 1 cm^2 bone, I've used 5 soaks of 30 - 60 minutes each. For fetal rat urogential sinus, 3 soaks of 2 to 3 minutes each works. For small (meaning a cm to mm) crustaceans and their appendages, which have relatively impermeable cuticles, I used 5 soaks of 5 or 10 minutes each. There can be some extraction of materials by the lq CO2 and possibly some artifacts from too long in lq CO2, but it is a much worse sin to run the specimens through CO2's critical point while they still retain EtOH. It is imperative to make certain that *all* of the EtOH has been replaced by lq CO2. This means you'll have to try different times and number of soaks. The larger and denser or woodier your samples, the more soaks for longer times you'll need. Some of the times I've given could perhaps be shortened. Given that too long likely won't produce significant artifact (note the "likely") and will only take more time, but that too little time will certainly produce ruined specimens which wastes a *lot* more time, specimens, and more, I'd rather spend the (possibly) extra time on the EtOH/lq CO2 exchanges.
Phil
} I am seeking advice on the correct way to operate a Tousimis } semiautomatic CPD to minimize if not eliminate } plant tissue shrinkage. } } I would appreciate any suggestions. } } Thank you. } } Greg Barclay
-- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
"There can be some extraction of materials by the lq CO2 and possibly some artifacts from too long in lq CO2...."
We are doing a student project here into SEM preparation methods for pollen. I wonder if you could let me know if you have any references for the above comment?
Thanks
Dave
On Sun, 20 Oct 2002 19:55:16 -0500 Philip Oshel {peoshel-at-facstaff.wisc.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Greg, } } This depends a great deal on the nature and size of the plant } tissues. We do a lot of small, herbaceous specimens here, mostly } Arabidopsis, but I've done other plant tissues as well, such as roots. } The trick is to ignore the directions in the Tousimis manual -- for } any biological specimens. What you need to do instead is to let the } specimens soak in the liquid CO2 for a given amount of time after } first purging the chamber of the absEtOH that was in it when the } samples were loaded. Then purge the chamber to replace the lq CO2 } with fresh lq CO2, soak, and repeat N times. This is analogous to the } dehydration process where the water in the tissues is replaced with } EtOH. Now, the EtOH in the tissues is being replaced with lq CO2. } Once that is done, then and only then can the samples be dried. } Otherwise, they are being dried from EtOH. } The length of the soaks and number of soaks is rather empirical and } highly dependent on tissue type and size. For the small (a few mm), } herbaceous Arabidopsis specimens, we generally use 4 soaks for 10 } minutes each. For 1 cm^2 bone, I've used 5 soaks of 30 - 60 minutes } each. For fetal rat urogential sinus, 3 soaks of 2 to 3 minutes each } works. For small (meaning a cm to mm) crustaceans and their } appendages, which have relatively impermeable cuticles, I used 5 } soaks of 5 or 10 minutes each. } There can be some extraction of materials by the lq CO2 and possibly } some artifacts from too long in lq CO2, but it is a much worse sin to } run the specimens through CO2's critical point while they still } retain EtOH. It is imperative to make certain that *all* of the EtOH } has been replaced by lq CO2. } This means you'll have to try different times and number of soaks. } The larger and denser or woodier your samples, the more soaks for } longer times you'll need. } Some of the times I've given could perhaps be shortened. Given that } too long likely won't produce significant artifact (note the } "likely") and will only take more time, but that too little time will } certainly produce ruined specimens which wastes a *lot* more time, } specimens, and more, I'd rather spend the (possibly) extra time on } the EtOH/lq CO2 exchanges. } } Phil } } } I am seeking advice on the correct way to operate a Tousimis } } semiautomatic CPD to minimize if not eliminate } } plant tissue shrinkage. } } } } I would appreciate any suggestions. } } } } Thank you. } } } } Greg Barclay } } -- } }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ } Philip Oshel } Supervisor, AMFSC and BBPIC microscopy facilities } Department of Animal Sciences } University of Wisconsin } 1675 Observatory Drive } Madison, WI 53706 - 1284 } voice: (608) 263-4162 } fax: (608) 262-5157 (dept. fax) }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
I had several very useful replies, thankyou to everyone who took the time to help. Although I am now able to connect and have the drive working on a WinNT PC, it appears that the disks are unreadable. The LEO 'scope has an operating system which pre-dates Win95 (even Win3.1, I think!), and the file allocation tables used on the MO disk can't be deciphered by the modern PC. So, it looks like I'll have to explore other routes. At the moment this looks like getting a CD-RW drive (if possible, I suspect there will be problems along this route too). I feel it should be possible to put a cable between adjacent boxes of electronics and send signals from one to another without the need for a physical medium inbetween, but maybe not...
======================================================================= This e-mail is intended for the person it is addressed to only. The information contained in it may be confidential and/or protected by law. If you are not the intended recipient of this message, you must not make any use of this information, or copy or show it to any person. Please contact us immediately to tell us that you have received this e-mail, and return the original to us. Any use, forwarding, printing or copying of this message is strictly prohibited.
No part of this message can be considered a request for goods or services. ======================================================================= Any questions about Bookham's E-Mail service should be directed to postmaster-at-bookham.com.
Le jeudi, 17 oct 2002, à 18:10 Europe/Paris, Dusevich, Vladimir a écrit :
} } We are going to move our EM lab to a new place } (in the same building) and are planning to put an } anti-vibration platform under the column of our } FEG-ESEM XL30. Do we have to place a HV transformer } on the same platform (the transformer is right behind } the column)?
Yes if the transformer is linked to the microscope with a rigid cable it would be betterto have it on the platform. Unless it generates by itself mechanical vibrations.
} } Any additional advise will be very helpful also, } especially experience with anti-vibration platforms } of different manufacturers.
We have recently built a platform for a new microscope. It is composed of a concrete block. I am sending you also pdfs of recent measurements. As you can see on the graphs we are rather satisfied of the results. The filtering system consists of a concrete bloc of 15 Mkg (tons) suspended with 4 springs with dampers at each corners. The company Gerb delivered the system. They made a calculation that the cutoff frequency would be around 2-3 Hz. Generally speaking such arrangement can be view as to spring in series (on for the block and one for the microscope). You must take care that there is no resonance between the to oscillators. In my case I made a quick estimate, which showed that this is not a problem. In fact as a rule of thumb the weight of the microscope should be negligible as compared to the platform. So design you platform with 10 times the weight of the microscope. The shape is also important. The vertical motion is normally the more important mode. But, if for example the shape of the concrete block is tall rather than flat, you may be annoyed by rotations around X, Y axis. This might excite flexion modes of the column, especially if their is some attachments like X-ray detector.. In our case we also have a preexisting concrete floor that is probably very thick but unfortunately connected to the building (old building). This probably why we can see the vibrations coming from the air conditioning which is located on the roof. If you make a new building you should consider building a concrete floor *de-connected* to the building and the your platform on dampers. Also, I have to mention that the operator is NOT on the platform but on a wooden floor above it. Finally, last advice and not the least: check EVERYTHING YOURSELF. Generally, the workers don't understand why you ask specific things and if they don't take care they can ruin your beautiful design. We had a mason who accidentally poured liquid concrete that you use to flatten the floors. Some of it went below the platform and re-connected it with the building!!! Fortunately if was not very difficult to remove. But what would happen if we haven't noticed?
Note about the graphs: please note that the vertical scales are correct. These measurements have been performed by our "vibration” department at Onera. Usually , they study vibrations modes of Aibuses and other flying machines. They had to build a special low noise amplifier for those measurements. This is the reason why we can see these very small contributions. I guess that with commercial equipments you would not be able to see these peaks above the noise. Best, Gilles
Our renal pathologist is moving to Columbus, Ohio and is taking a position at Ohio State. He is looking for a trained EM tech who is experienced in renal pathology EM. Contact me off list directly to my email address if anyone is interested.
karen_bentley-at-urmc.rochester.edu
Karen
Karen L. Bentley, M.S.(previously Jensen) Associate Scientist & Project Manager Electron Microscope Research Core University of Rochester Medical Center Rochester, NY 14642 585-275-1954
Dear all, What are the alternatives to C-support films, for supporting powder particles. We have a situation where we want to determine the C content of an inorganic phase and don't want a C film, which would mess up the quantification.
Hello again. Sorry to hear about the difficulties.
The first order of business will be to find out what system is running the LEO. After that is done, you can probably explore options. They may not all be that much fun. There is a possibility that there will not only be an operating system incompatibility, but also a file format compatibility.
We were running a Kevex EDS system when the Internet started coming on big. We much wanted to make files available over the net - me especially because I had my office in another building. The Kevex operated around a DEC PDP-11 computer with the TSX+ operating system. (Those aren't exactly common.) The first order of business was to connect that to the ethernet. We were able to do it but it required a special card costing several hundred dollars (now you can buy $10 ethernet cards for PCs) and TCP-IP software costing many hundreds more (compared to the built in support in modern OSs). Then we had to convert the proprietary Kevex file format over to a portable format (TIF). Once we had crossed those two not-so-small hurdles, we were up and running just fine.
The other approach, like you have been trying, was to find a common media format so that we could transfer removable media between two computers. Someone had written a utility to read PDP-11 disks on a PC (getting around the different allocation and directory structures), but I think it may have been limited to floppy disks. We were using 10 MB Bernoulli cartridges from Iomega. I vaguely recall that there were other issues trying to access those disks. Even if we had succeeded, there would still have been the issue of converting the file formats.
I would like to think that it will not be so bad for you. I would like to think that the LEO is built around a PC. But LEO or another LEO user would probably have to verify that. They should also be able to tell which OS and what disk system are being used. Then you could move on from there.
Warren
At 11:06 AM 10/21/02 +0100, you wrote:
} I had several very useful replies, thankyou to everyone who took the time to } help. } Although I am now able to connect and have the drive working on a WinNT } PC, it appears that the disks are unreadable. The LEO 'scope has an } operating system which pre-dates Win95 (even Win3.1, I think!), and the file } allocation tables used on the MO disk can't be deciphered by the modern PC. } So, it looks like I'll have to explore other routes. At the moment this } looks like getting a CD-RW drive (if possible, I suspect there will be } problems along this route too). I feel it should be possible to put a cable } between adjacent boxes of electronics and send signals from one to another } without the need for a physical medium inbetween, but maybe not... } } Thanks again, } } Richard
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
5116-207GE M:1516 E 0065 + G:15/3 (reduction ratio 900:1) and 5116-211GE M:1516 E 0065 + G:15/3 (reduction ratio 1670:1)
They are need for positioning and rotation of specimen table in BALZERS Universal Ion Beam Etching Device IEU 100.
With kind regards, Dmitri Lobanov.
Institute for physics of microstructures RAS 603950, N.Novgorod, GSP-105, Russia phone: +7(8 312)675037 fax: +7(8 312)609111 E-mail: dima-at-ipm.sci-nnov.ru
I am looking for a lab that has a tensile stage mounted in an ESEM that is willing to run a couple of samples. I would prefer a service lab, however if there is a commercial or university lab that is willing to take on the task, that would be fine as well. The samples would be polymer films and I would be interested in images corresponding to the various force data. If you have this capability, please contact me off list.
Thanks in advance,
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108 David_Bell-at-Millipore.com
Does anyone know a source for a variable temperature hot plate? We want to use it for wax embedding, for pouring in situ boats, allowing us to slide em over and readjust the tissue as it solidifies.
Is anyone interested in a diagnostic EM job? You must have experience in renal pathology and tumors. Our renal pathologist is moving to Ohio and is looking for anyone in the USA who is interested and available.
Please contact me off list at: karen_bentley-at-urmc.rochester.edu
Karen L. Bentley, M.S.(previously Jensen) Associate Scientist & Project Manager Electron Microscope Research Core University of Rochester Medical Center Rochester, NY 14642 585-275-1954
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: White, Woody N. [mailto:nwwhite-at-mcdermott.com] Sent: Monday, October 21, 2002 7:59 AM To: 'Diane G. Miller'; microscopy-at-sparc5.microscopy.com
Most image editors will let you save images in a multitude of formats. I your case it would be load-resave steps. Some may allow batch processing.
Examples: IrView http://www.irfanview.com/ Or VuePrint http://www.hamrick.com/
Regards, Woody
Woody White McDermott Technology Inc. McD: http://www.mtiresearch.com/ Mine: http://woody.white.home.att.net
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello All,
I was wondering if some of you could forward me some sites for converting bitmap images to tiff or jpeg. I would appreciate it very much.
Thanks for your help. Diane
Diane G. Miller Miller Consultant Service 503-784-6444 millerd-at-coho.net
Hi, I am posting this message for Prof. Dravid. Please contact him directly. Thank you, Shuyou. ======================================================= IMMEDIATE POSTDOCTORAL POSITION AVAILABLE:
CRYO-ANALYTICAL TRANSMISSION ELECTRON MICROSCOPY OF SOFT AND HYBRID NANOSTRUCTURES
INSTITUTE FOR ENVIRONMENTAL CATALYSIS, NORTHWESTERN UNIVERSITY
Applications are invited for a full-time postdoctoral research associate within the Institute for Environmental Catalysis (IEC) at Northwestern University. The position is jointly supervised by Profs. Vinayak P. Dravid of Materials Science & Engineering, and Prof. Jean-Francois Gaillard of Civil Engineering, at Northwestern University.
We seek a dynamic and energetic individual with prior experience in advanced electron microscopy of micobiological, geochemical, or materials systems to conduct frontier research using state-of-the-art TEM instrumentation; and design novel approaches to observe aquatic environmental particles in-situ in TEM. Hands-on experience in preparatory methods such as cryo-preparation, ultramicrotomy etc. will be a strong asset, as well as knowledge of STEM-EDS, PEELS, and energy filtering.
Applications should include a vita, a brief statement of research interests, and names of at least three references including email addresses and telephone numbers.
The position is open only to those eligible for employment in the US.
Applications in PDF format should be sent by email to: v-dravid-at-northwestern.edu
Professor Vinayak P. Dravid, Materials Science & Engineering, and Institute for Environmental Catalysis 2225 N. Campus Drive, 1133 Cook Hall (formerly MLSF) Northwestern University, Evanston, IL 60208, USA Ph.: (847) 467-1363, Fax: (847) 467-6573 E-mail: v-dravid-at-northwestern.edu http://vpd.ms.northwestern.edu http://epic.ms.northwestern.edu
_____________________________ Shu-You Li, Ph.D. Electron Microscopist Electron Probe Instrumentation Center(EPIC) Northwestern University 2220 Campus Drive, 1141 Cook Hall Evanston, IL 60208, USA Ph: (847) 491-7798, Fax: (847) 491-7820 Email: syli-at-northwestern.edu; syli16-at-hotmail.com http://pubweb.northwestern.edu/~sli974
Ian MacLaren wrote: ========================================================= What are the alternatives to C-support films, for supporting powder particles. We have a situation where we want to determine the C content of an inorganic phase and don't want a C film, which would mess up the quantification. ========================================================== There are two possibilities:
a) silicon dioxide/monoxide films made via the evaporation of SiO, see URL http://www.2spi.com/catalog/spec_prep/evapor_3b.shtml or the completed films can be purchased already made from SPI Supplies (and some of our competitors), see URL http://www.2spi.com/catalog/grids/cusctgrd.html
or
b) silicon nitride membrane window TEM grids as shown on URL http://www.2spi.com/catalog/instruments/silicon-nitride.shtml
Either way, you will have a good support that will be completely free of any carbon.
Disclaimer: SPI Supplies provides all three items mentioned, namely the SiO chips, already custom coated SiO2 TEM grids, and the silicon nitride membrane window grids.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
If anyone has any information about labs that do contract micro-FTIR work in the Texas area, please forward it to me. We are looking for a source for this type of work closer than Sunnyvale, CA.
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-648-8743 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
A TEM Specimen Preparation Course Will be offered March 12-14, 2003 at the Materials Characterization Facility at the University of Central Florida, Orlando, FL USA
A review of all techniques will be given, with primary emphasis on tripod polishing, ion milling, and all latest FIB techniques
Instructors: Ron Anderson, Microscopy Today Fred Stevie, NC State Lucille Giannuzzi, UCF Brian Kempshall, UCF
For more information contact Lucille Giannuzzi, lag-at-mail.ucf.edu
******************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor, Mechanical, Materials, and Aerospace Engineering Director, UCF/Cirent Materials Characterization Facility University of Central Florida 12443 Research Parkway, Suite 305 Orlando, FL 32826 email lag-at-mail.ucf.edu phone (407) 882-1500 fax (407) 275-4321 -------------------------------------------------------------------- "Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain ********************************************************************
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (pkrsko-at-stevens-tech.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, October 21, 2002 at 13:31:19 ---------------------------------------------------------------------------
Email: pkrsko-at-stevens-tech.edu Name: Peter Krsko
Organization: Stevens Institute of Technology
Education: Graduate College
Location: Hoboken, NJ 07030
Question: Dear Sirs,
Where can I purchase some kind of calibration sample for fluorescent imaging? I would like to qauntify my results, but I am not sure what intensity corresponds to what concentration of FITC. Are there also calibration samples for reflective fluoro microscopy?
CALL FOR PAPERS FIB Focused Interest Group (FIG) WORKSHOP
Tuesday, March 18, 2003
Student Union, University of Central Florida, Orlando, FL 32816
(in conjunction with a TEM Specimen Prep Course March 12-14, 2003, and the FL AVS/Florida Society for Microscopy Meeting, March 17-18, 2003).
Please submit an abstract (limit 250 words), including authors and affiliations, by email to:
Dr. Lucille Giannuzzi lag-at-mail.ucf.edu
******************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor, Mechanical, Materials, and Aerospace Engineering Director, UCF/Cirent Materials Characterization Facility University of Central Florida 12443 Research Parkway, Suite 305 Orlando, FL 32826 email lag-at-mail.ucf.edu phone (407) 882-1500 fax (407) 275-4321 -------------------------------------------------------------------- "Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain ********************************************************************
I would appreciate recieveing any fixation protocol you know works well for mouse muscle (extensor digitorum longus (EDL) and epitrochlearus (EPI)). We use routinely immersion into 2.5% glutaraldehyde in 0.1M cacodylate pH 7.4, but have had often problems with mitochondrial swelling. Since this project is to quantify mitochondrial density, we do need perfect and reliable fixation of all mitos. Perfusion is not an option. Thanks for any advise on type of fixative, buffer and temperature.
Marc -- Marc Pypaert Department of Cell Biology, Center for Cell and Molecular Imaging and Ludwig Institute for Cancer Research, Yale University School of Medicine 333 Cedar Street New Haven CT 06520 Tel : (203) 785 3681 Fax : (203) 785 7446
Why don't you embed the samples in glass? You are at the softening temperatures of glass. We have a technique here that we bond two pieces of glass together by heating them in a plasma etching system. This technique was originally developed and published by Prof Carlos Pantano at Penn State University to profile Sn on the bottom surface of glass. What I would do is sprinkle some of your dust on one of the glass surfaces, bring the two glass surfaces together and heat them up. Then dimple and ion mill normally. I have no idea whether you particles will wet the glass or not, but they might.
Another option is to use a ceramic bonding adhesive in a similar manner to above. There is a description of a high temperature XTEM prep in the MRS TEM sample Prep book number 1 on what to use. The adhesive was from Aremco. Take two piece of Si and put your adhesive on one surface, dust the adhesive and make your sandwich. Alternatively, you could just embed them in the ceramic adhesive and prepare the disk as a plan view sample.
If you choose one of these ideas, I would love to know how it works out.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
-----Original Message----- } From: "George_Munzing-at-engelhard.com"-at-sparc5.microscopy.com [mailto:"George_Munzing-at-engelhard.com"-at-sparc5.microscopy.com] Sent: Tuesday, October 15, 2002 9:23 AM To: Microscopy-at-sparc5.microscopy.com
Hello fellow microscopists,
I need to do some high temperature TEM studies and am looking for a suitable mounting resin. I will be embedding and microtoming powder materials and would like to go as high in temperature as possible. The materials themselves can withstand the temperatures but I doubt my conventional epoxy would survive.
Is there any silicone epoxy or other suitable material designed to withstand high temperatures (maybe as high as 600-800C) for a short time for embedding?
Any suggestions you can offer are greatly appreciated.
Thanks in advance!
George R. Munzing Jr. Engelhard Corporation Strategic Technologies Group 25 Middlesex/Essex Turnpike Iselin, NJ 08830 Tel: 732-205-7030 FAX: 732-205-5300
Cole-Parmer lists temperature-controlled hot plates (made by Heisen, and probably others)--from fixed setpoint units with +/-1 degree temperature control to fancy programmables. I'm pretty sure that the other major lab supply houses (Fisher, VWR, etc.) also offer variable-temperature hot plates, but I don't have catalogs handy at the moment, so I can't verify that.
You might also consider using a Variac(TM) (or other variable autotransformer) to power a basic, old-fashioned clunker of a hot plate. If you use that approach, make sure the power capacity of the autotransfomer matches the power of the hot plate. Don't buy a Variac that would let users dial in too high a voltage, which could fry the hot plate and/or present safety hazards. And don't use the transformer approach if your hot plate has fancy extras such as stirmotors, digital displays, etc.---many devices other than plain-vanilla electrical resistance heaters can be damaged by undervoltage. Also, the autotransformer strategy would require that you invest some time to "calibrate" your setup: i.e,, to find out at what voltage the hot plate maintains a satisfactory and safe temperature for what you want to do. Changing the environment or setup (anything that increases or decreases heat losses from the hot plate) would Furthermore, autotransformers are pricey items (as are T-controlled hot plates).
So your best bet, probably, is to buy a nice temperature-controlled hot plate, as you indicated.
Hope this helps.
-------Roy
P.S. I have no connection to any of the suppliers or manufacturers mentioned, and no preference for any supplier is intended. Shop with care to make sure anything you buy meets your requirements.
====================== Roy Arrowood arrowood-at-utep.edu Metallurgical and Materials Engineering University of Texas at El Paso El Paso, TX 79968-0520 (915)747-6934 voice // (915)747-8036 FAX (915)747-5468 secretary
Does anyone have a method or protocol for synchronizing cells in tissue culture?
We are looking to find a method so that all cells in the culture are at substantially the same point in their cell cycle? It is also important that the cells are still alive and living as normally as cells do when in tissue culture.
We are most interested in achieving this goal with human fibroblasts in culture, but would be interested in any success with other cells lines of any origin, mammal, animal, or plant.
Thank you for your assistance, any information on achieving this goal will be helpful.
Tim Richardson, RTI, Phone: 905-951-7058 Fax: 905-951-7052
"There can be some extraction of materials by the lq CO2 and possibly some artifacts from too long in lq CO2...."
We are doing a student project here into SEM preparation methods for pollen. I wonder if you could let me know if you have any references for the above comment?
Thanks
Dave
On Sun, 20 Oct 2002 19:55:16 -0500 Philip Oshel {peoshel-at-facstaff.wisc.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Greg, } } This depends a great deal on the nature and size of the plant } tissues. We do a lot of small, herbaceous specimens here, mostly } Arabidopsis, but I've done other plant tissues as well, such as roots. } The trick is to ignore the directions in the Tousimis manual -- for } any biological specimens. What you need to do instead is to let the } specimens soak in the liquid CO2 for a given amount of time after } first purging the chamber of the absEtOH that was in it when the } samples were loaded. Then purge the chamber to replace the lq CO2 } with fresh lq CO2, soak, and repeat N times. This is analogous to the } dehydration process where the water in the tissues is replaced with } EtOH. Now, the EtOH in the tissues is being replaced with lq CO2. } Once that is done, then and only then can the samples be dried. } Otherwise, they are being dried from EtOH. } The length of the soaks and number of soaks is rather empirical and } highly dependent on tissue type and size. For the small (a few mm), } herbaceous Arabidopsis specimens, we generally use 4 soaks for 10 } minutes each. For 1 cm^2 bone, I've used 5 soaks of 30 - 60 minutes } each. For fetal rat urogential sinus, 3 soaks of 2 to 3 minutes each } works. For small (meaning a cm to mm) crustaceans and their } appendages, which have relatively impermeable cuticles, I used 5 } soaks of 5 or 10 minutes each. } There can be some extraction of materials by the lq CO2 and possibly } some artifacts from too long in lq CO2, but it is a much worse sin to } run the specimens through CO2's critical point while they still } retain EtOH. It is imperative to make certain that *all* of the EtOH } has been replaced by lq CO2. } This means you'll have to try different times and number of soaks. } The larger and denser or woodier your samples, the more soaks for } longer times you'll need. } Some of the times I've given could perhaps be shortened. Given that } too long likely won't produce significant artifact (note the } "likely") and will only take more time, but that too little time will } certainly produce ruined specimens which wastes a *lot* more time, } specimens, and more, I'd rather spend the (possibly) extra time on } the EtOH/lq CO2 exchanges. } } Phil } } } I am seeking advice on the correct way to operate a Tousimis } } semiautomatic CPD to minimize if not eliminate } } plant tissue shrinkage. } } } } I would appreciate any suggestions. } } } } Thank you. } } } } Greg Barclay } } -- } }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ } Philip Oshel } Supervisor, AMFSC and BBPIC microscopy facilities } Department of Animal Sciences } University of Wisconsin } 1675 Observatory Drive } Madison, WI 53706 - 1284 } voice: (608) 263-4162 } fax: (608) 262-5157 (dept. fax) }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Marc, We use to routinely fix mouse muscle by immersion but used 3.5% glut. in .1M cacodylate at 4 degrees for 1.5 hours. The pieces need to be very small. Add 4% sucrose to the buffer washes. The pH should be 7.4. Osmium should be 1% in the same buffer at 4 degrees also. Good luck, Mary Gail
At 04:48 PM 10/21/02 -0600, Marc Pypaert wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Mary Gail Engle Sr. Research Laboratory Manager Electron Microscopy & Imaging Facility Health Sciences Research Bldg. 001 University of Kentucky Lexington, KY 40536-0305
Hi Marc, I've always had goo results in muscle (skeletal and cardiac) with the following fix: 4% pfa (methanol-free, diluted from 16% stock ) 2.5% glutaraldehyde in 0.1M Na-cacodylate buffer My "recipe", making this up from 10ml stock vials of 16% pfa and 10% glut yields 40 ml of fix, to which I then add 2 ml of saturated picric acid (aqueous). (based on Ito & karnovsky, J Cell Bio, abstract 168a, 1968)
Try to hold the muscles in an extended state, either by leaving them attached to the leg bone, or using ligature to tie them to small wooden sticks. This will help prevent overly contracted sarcomeres and give a more "textbook" appearance with M-lines, A & I bands, etc. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
An associate of mine is looking for a service lab in Taiwan that would be able to pot and polish samples and examine the samples with a SEM. Any information as to company names and/or contacts would be appreciated.
There is at least one study showing that the important thing, as far a shrinkage or swelling is concerned, is the osmolarity of the buffer. This should be close to physiological (300 mOsm). Glutaraldehyde will add about 100 mOsm for each percent, thus, a 1% glutaraldehyde solution in buffer that would be 300 mOsm by itself would be ~400 mOsm. However, the tonicity added by the fixaive apparently doesn't detract from the morphology.
The 0.1 M cacodylate + 4% sucrose mentioned below will be close to the percent we used to use with good success. I think, however, our final sucrose concentration was 3.8%. We used to make up double strength buffer and double strength aqueous fix and add them together 1:1; e.g., 0.2 M cacodylate containing 6.8% sucrose and 4 % aqueous glut. Our final concentration after mixing them 1:1 was 0.1 M caco, 3.4% sucr, 2% glut. I think 3.5% glut is slight overkill, but won't hurt anything. Some folks use 5%. You could also make the buffer with an increased concentration of cacodylate, but it is expensive. 1M is sufficient buffering capacity, and the sucrose makes up the rest of the tonicity.
Cacodylate will give a finer grained ultrastructure than phosphate, but some folks don't want to use the arsenical. If you choose phosphate, 0.135 M phosphate buffer is physiological; you can also use less phosphate and make up the difference with cheap sucrose. If you use something that is ionic, like NaCl, remember that it dissociates into 2 ions and you have to use half as much. Also, the 3.4 % sucrose translates in molarity to 0.1 M, thus if you use an ionic substitute, use molarity to calculate the amount, not percent. A 1M solution of sucrose and a 0.5 M solution of NaCl will give the same osmolarity.
Additionally, a small amount (2.5 mM, i.e., 0.0025 M) of calcium is good for membranes. This is not enough to alter the osmolarity very much.
The other thing you need to do if you're interested in muscle bands (and maybe mitochondrial size???) is to fasten the muscle bundle in a muscle clamp before immersing in fix. Otherwise, it contracts.
This may be more than you wanted to know, but hope some of it helps.
Sara Miller
On Tue, 22 Oct 2002, Mary Gail Engle wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Marc, } We use to routinely fix mouse muscle by immersion but used 3.5% glut. in } .1M cacodylate at 4 degrees for 1.5 hours. The pieces need to be very } small. Add 4% sucrose to the buffer washes. The pH should be 7.4. Osmium } should be 1% in the same buffer at 4 degrees also. } Good luck, } Mary Gail } } At 04:48 PM 10/21/02 -0600, Marc Pypaert wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Dear List, } } } } I would appreciate recieveing any fixation protocol you know } } works well for mouse muscle (extensor digitorum longus (EDL) } } and epitrochlearus (EPI)). We use routinely immersion into } } 2.5% glutaraldehyde in 0.1M cacodylate pH 7.4, but have had } } often problems with mitochondrial swelling. Since this project is } } to quantify mitochondrial density, we do need perfect and } } reliable fixation of all mitos. Perfusion is not an option. Thanks } } for any advise on type of fixative, buffer and temperature. } } } } Marc } } -- } } Marc Pypaert } } Department of Cell Biology, } } Center for Cell and Molecular Imaging and } } Ludwig Institute for Cancer Research, } } Yale University School of Medicine } } 333 Cedar Street } } New Haven CT 06520 } } Tel : (203) 785 3681 } } Fax : (203) 785 7446 } } } } } } } Mary Gail Engle } Sr. Research Laboratory Manager } Electron Microscopy & Imaging Facility } Health Sciences Research Bldg. 001 } University of Kentucky } Lexington, KY 40536-0305 } } phone 859-323-6108 } fax 859-257-9700 } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
From daemon Tue Oct 22 11:00:39 2002
Contents Retrieved from Microscopy Listserver Archives