} ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } After years of trying desperately to get one of our microtomes } working, I'm thinking of changing the company that provides } service on our instruments (2 Reichert Ultracut E and 2 Leica } UCT/FCS). Does anyone in the NorthEast know of someone } capable of providing service in Connecticut? I'm especially } interested in someone with good experience with the } Leica instruments. } Thank you } } Marc } -- } Marc Pypaert } Department of Cell Biology } Center for Cell and Molecular Imaging } Ludwig Institute for Cancer Research } Yale University School of Medicine } 333 Cedar Street, PO Box 208002 } New Haven, CT 06520-8002 } TEL 203-785 3681 } FAX 203-785 7446 } }
-- -- ********************************************** 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 **********************************************
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 1 08:52:21 2004
I am also interested in getting a repairperson for my Ultracut E. It really needs an overhaul.
Peggy
Peggy Sherwood Lab Associate, Photopathology Wellman Center for Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
-----Original Message----- } From: Marc Pypaert [mailto:marc.pypaert-at-yale.edu] Sent: Thursday, September 30, 2004 6:58 PM To: 'Microscopy-at-MSA.Microscopy.Com'
After years of trying desperately to get one of our microtomes working, I'm thinking of changing the company that provides service on our instruments (2 Reichert Ultracut E and 2 Leica UCT/FCS). Does anyone in the NorthEast know of someone capable of providing service in Connecticut? I'm especially interested in someone with good experience with the Leica instruments. Thank you
Marc -- Marc Pypaert Department of Cell Biology Center for Cell and Molecular Imaging Ludwig Institute for Cancer Research Yale University School of Medicine 333 Cedar Street, PO Box 208002 New Haven, CT 06520-8002 TEL 203-785 3681 FAX 203-785 7446
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 1 09:03:25 2004
I use Microscopical Optical Consulting, out of Valley Cottage NY. You can reach them at 845-268-6450.
Ann Lehman Electron Microscopy Facility Mailstop: LSC314 Trinity College 300 Summit Street Hartford, CT 06106 v. 860-297-4289 f. 860-297-2538 e. ann.lehman-at-trincoll.edu w. http://www.trincoll.edu/~alehman
-----Original Message----- } From: Sherwood, Margaret [mailto:MSHERWOOD-at-PARTNERS.ORG] Sent: Friday, October 01, 2004 9:47 AM To: 'Marc Pypaert'; 'Microscopy-at-MSA.Microscopy.Com'
Marc, et al,
I am also interested in getting a repairperson for my Ultracut E. It really needs an overhaul.
Peggy
Peggy Sherwood Lab Associate, Photopathology Wellman Center for Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
-----Original Message----- } From: Marc Pypaert [mailto:marc.pypaert-at-yale.edu] Sent: Thursday, September 30, 2004 6:58 PM To: 'Microscopy-at-MSA.Microscopy.Com'
After years of trying desperately to get one of our microtomes working, I'm thinking of changing the company that provides service on our instruments (2 Reichert Ultracut E and 2 Leica UCT/FCS). Does anyone in the NorthEast know of someone capable of providing service in Connecticut? I'm especially interested in someone with good experience with the Leica instruments. Thank you
Marc -- Marc Pypaert Department of Cell Biology Center for Cell and Molecular Imaging Ludwig Institute for Cancer Research Yale University School of Medicine 333 Cedar Street, PO Box 208002 New Haven, CT 06520-8002 TEL 203-785 3681 FAX 203-785 7446
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 1 15:02:20 2004
We just installed a new TEM and it was suggested that we use a 'demand regulator' to control the flow of nitrogen during specimen exchange and/or venting the column etc. The installing engineer didn't have the details for ordering etc.
Ok, so tell me about demand regulators and where to get one.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 1 18:15:35 2004
Our Mini-Workshop Series on "Cryoultramicrotomy for Materials Science" is coming to Akron, Ohio!
**When**: Thursday, October 14 - Friday, October 15, 2004, 9:00am-4:00pm
**Where**: University of Akron Department of Polymer Engineering Polymer Engineering Academic Center (PEAC) Room 307 250 South Forge Street Akron, OH 44325
**What**: A two-day mini-workshop with presentations, demonstrations and hands-on sessions for cryoultramicrotomy, including toolmaking (wire loops and hair probes), glass knife making, evaluation of glass knives, care and cleaning of diamond knives and operation of the cryoultramicrotome.
Lectures, demonstrations and an introduction to the cryoultramicrotome will be given on the first day; open lab sessions and hands-on use of the instrumentation in small groups will be the focus of the second day.
Demonstrations will be given using well-characterized specimens which are provided by the instructors. There is a limited amount of time to work with specimens provided by attendees. If you would like to bring your own specimens, please provide details on the materials when you RSVP and reserve a place in the workshop.
**Background**: These techniques are of special interest to anyone working with polymers or other materials which could benefit from sectioning or surface "polishing"at low temperatures (no embedding required). The sections may be used for TEM, optical microscopy, IR spectroscopy and FTIR. The polished surface of the bulk material is suitable for AFM, SEM, X-ray microanalysis and elemental mapping.
**Important Info**:
1. There is no charge for this workshop.
2. Meals and refreshments will be served!
3. Attendance is open to everyone for the presentations and demonstrations on the first day. However, attendance is limited for the cryoultramicrotomy hands-on sessions on the second day.
**RSVPs and Reservations**: To RSVP and to reserve a spot for the hands-on sessions, please contact Kim Megaw at Boeckeler Instruments, Inc. ( {kim-at-boeckeler.com} , 800.552.2262).
**Sponsors and Organizers**: University of Akron Department of Polymer Engineering RMC Products Group, Boeckeler Instruments, Inc.
See you in Akron!
Bob Chiovetti Senior Product Specialist Boeckeler Instruments, Inc.
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 1 20:41:20 2004
Check your local scuba shop. I used some PVC (Tygon) tubing, a few hose clamps, and a few copper plumbing fittings to adapt a scuba regulator for my dry nitrogen backfill.
I hose-clamped a piece of large diameter Tygon over the mouthpiece then downsized with the Cu fittings to a 1/4" hose barb which hooked to the hose to the scope. On the supply side, I cut the original fitting off the high pressure hose and used a brass hose barb to connect it to my nitrogen gas regulator.
Have fun explaining to your financial people why you needed to buy a scuba regulator!
Cheers, Henk
At 04:03 PM 10/1/2004, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hendrik O. Colijn colijn.1-at-osu.edu OSU Campus Electron Optics Facility www.ceof.ohio-state.edu 040 Fontana Labs, 116 W. 19th Ave
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 2 03:31:27 2004
Why is this stuff not being supplied by the vendor, I wonder.
Chris Jeffree University of Edinburgh
----- Original Message ----- From: "Jon Krupp" {jmkrupp-at-cats.ucsc.edu} To: {Microscopy-at-microscopy.com} Sent: Friday, October 01, 2004 9:03 PM Subject: [Microscopy] Demand regulator
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hi: } } We just installed a new TEM and it was suggested that we use a } 'demand } regulator' to control the flow of nitrogen during specimen exchange } and/or } venting the column etc. The installing engineer didn't have the } details for } ordering etc. } } Ok, so tell me about demand regulators and where to get one. } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } } }
From MicroscopyL-request-at-ns.microscopy.com Sun Oct 3 09:40:29 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ghina_24-at-yahoo.com) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Sunday, October 3, 2004 at 00:06:00 ---------------------------------------------------------------------------
Tina, I recently fixed yeast cells for the first time and also found that it is hard to see the membranes. I followed a protocol suggested by Mark Winey which referenced a paper by Byers (Byers, B. and Goetsch, L. (1991) Preparation of yeast cells for thin section electron microscopy. Methods Enzymol. 194: 602-608.) This procedure involves spheroplasting which removes the cell wall, thus allowing good infiltration. The cells turned out looking great but the people I did the work for wanted to see more membranes.
Another good reference for fixing yeast is a paper by Robin Wright (Wright,R. (2000) Transmission Electron Microscopy of Yeast. Microsc. Res. Tech. 51:496-510). She gives alot of helpful hints for fixing yeast. She gives a detailed procedure in the paper using potassium permanganate as a post fixative. If you use potassium permanganate as a post fixative you don't have to spheroplast the cells (according to the paper, I have not tried it yet). Of course we all know that potassium permanganate extracts much of the cytoplasm, which is perfect for viewing membranes. I hope this will be helpful.
Janice Pennington Senior Electron Microscopist Nephrology/Anatomy and Cell Biology Indiana University School of Medicine 635 Barnhill Drive, MS 5065 Indianapolis, IN 46202 Phone (317) 274-8730 Fax (317) 278-2040
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 4 09:10:21 2004
I would like to open a discussion about the use of Biosafety Level 2 regulated cells, tissues and organisms in a multi-user microscopy facility not designed to house BSL2 regulated agents (no once-through air or biosafety cabinet).
Is it customary to have researchers use the microscopy facility if BSL2 regulated agents are alive but completely contained in unbreakable and well-sealed containers? While such conditions are easily accomplished for transport of materials, they are less so for the actual viewing. How is this commonly resolved?
Is it customary for users to write a protocol for microscope use, spelling out what to do in an emergency situation?
Is expected from microscopy facility personnel to know exactly what to do in case of an emergency situation (e.g. if somebody works on BSL2 regulated bacteria in a flow cell and one of the connections breaks during microscope operation, spilling the content over microscope equipment), or is this the responsibility of the investigator?
In case of spillage, a broken slide, etc., how does one deal with the resulting contaminated equipment?
Is it customary to have every user fill out and sign a form explaining what organisms they plan to use, how it is regulated and how they plan to use those organisms in the microscopy facility?
Thank you for your input.
All the best, Judith.
Judith Wopereis Microscopy Facility Manager and Instructor of Laboratories in Biological Sciences Smith College Northampton, MA 01063 (413) 585 3829 (voice mail) (413) 585 3786 (fax) jwoperei-at-email.smith.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 4 09:12:36 2004
We purchased ours through our local compressed gas vendor.
Hendrik O. Colijn wrote:
} } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } Jon, } } Check your local scuba shop. I used some PVC (Tygon) tubing, a few } hose clamps, and a few copper plumbing fittings to adapt a scuba } regulator for my dry nitrogen backfill. } } I hose-clamped a piece of large diameter Tygon over the mouthpiece } then downsized with the Cu fittings to a 1/4" hose barb which hooked } to the hose to the scope. On the supply side, I cut the original } fitting off the high pressure hose and used a brass hose barb to } connect it to my nitrogen gas regulator. } } Have fun explaining to your financial people why you needed to buy a } scuba regulator! } } Cheers, } Henk } } } At 04:03 PM 10/1/2004, you wrote: } } } } ------------------------------------------------------------------------------ } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------------------------- } } } } } } Hi: } } } } We just installed a new TEM and it was suggested that we use a 'demand } } regulator' to control the flow of nitrogen during specimen exchange } } and/or } } venting the column etc. The installing engineer didn't have the } } details for } } ordering etc. } } } } Ok, so tell me about demand regulators and where to get one. } } } } Thanks } } } } Jonathan Krupp } } Microscopy & Imaging Lab } } University of California } } Santa Cruz, CA 95064 } } (831) 459-2477 } } jmkrupp-at-cats.ucsc.edu } } } Hendrik O. Colijn colijn.1-at-osu.edu } OSU Campus Electron Optics Facility www.ceof.ohio-state.edu } 040 Fontana Labs, 116 W. 19th Ave } } }
-- -- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 4 10:14:05 2004
I would like to know if there is a mixture of epoxy and carbon powders for ion- thinning in market. A researcher from Hungary made a low angle ion-thinner (IV3?), and also he confirmed that if you use C powder with epoxy, you can ion- thin fine grain materials like powder. I do not remember the reference now, but you can get a lower thinning rate of the C and epoxy mixture, according to the paper. I used a kit for ion-thinning in Japan. Probably, it was imported from Hungary. If you use similar technique, please advise where I can get the powdered epoxy and C.
Thank you, Hiromi Konishi, Ph.D. Indiana University
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 4 10:29:26 2004
We have also used these on several instruments at our facility. Alex Greene set them up. The only difference was he filled the mouthpiece with RTV sealant and used a secondary output on the regulator to connect to the scope. I am not sure if all SCUBA regulators have this output, or if the ones he found were special.
One problem you might find is if the demand is too high or the supply too low, the small silicone diaphragm used for exhaling can invert allowing room air into the system. I have also always worried (however with no proof) that the line between the regulator and the instrument, being at zero pressure WRT the room, can become contaminated with room air.
On our latest TEM and SEM installs, we have not used these since the manufactures have included doors or popup valves that open when the pressure difference becomes zero WRT the room.
-Ray
Ray D. Twesten, PhD. Center for Microanalysis of Materials Seitz Materials Research Laboratory 104 S. Goodwin Ave., Urbana, IL 61801 +1 217 244-6177 (Fax -2278)
-----Original Message----- } From: colijn-at-er6s1.ecr6.ohio-state.edu [mailto:colijn-at-er6s1.ecr6.ohio-state.edu]On Behalf Of Hendrik O. Colijn Sent: Friday, October 01, 2004 8:47 PM To: Microscopy-at-microscopy.com
Jon,
Check your local scuba shop. I used some PVC (Tygon) tubing, a few hose clamps, and a few copper plumbing fittings to adapt a scuba regulator for my dry nitrogen backfill.
I hose-clamped a piece of large diameter Tygon over the mouthpiece then downsized with the Cu fittings to a 1/4" hose barb which hooked to the hose to the scope. On the supply side, I cut the original fitting off the high pressure hose and used a brass hose barb to connect it to my nitrogen gas regulator.
Have fun explaining to your financial people why you needed to buy a scuba regulator!
Cheers, Henk
At 04:03 PM 10/1/2004, you wrote:
} --------------------------------------------------------------------------- --- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hendrik O. Colijn colijn.1-at-osu.edu OSU Campus Electron Optics Facility www.ceof.ohio-state.edu 040 Fontana Labs, 116 W. 19th Ave
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 4 11:44:37 2004
First of all, I can vouch for the Wright protocol for preparing yeast for TEM mentioned by Janice (1. below). Works great! Membranes stand right out there, resins infiltrate through wall completely into cell (I used Embed812).
In your query (2. below) you mention that the ultimate goal of your study is to look for copper in mitochondria in yeast, using EELS or whatever analytical technique. For elemental analysis in sections, given all the prep they usually go through, there is always the danger of leaching out the element you are interested in looking for. Any elements/compounds that are water soluble - they're gone! So even if you succeed in getting good membrane and mitochondrial fixation, may not be any copper left. Do you know for sure if the copper is bound in the mitochondria?
I have heard that freeze-substitution does a better job of preserving elements in place for elemental analysis - you'd just have to try it and see. Or try freeze-drying small masses of sample, then vacuum infiltrate with resin.
Good luck! Gib -- Gib Ahlstrand, Scientist Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)624-2785 FAX, ahlst007-at-tc.umn.edu http://www.cbs.umn.edu/ic/
"You can learn a lot by observation - just by lookin'!" - Yogi Berra -------------
1. --------- Tina, I recently fixed yeast cells for the first time and also found that it is hard to see the membranes. I followed a protocol suggested by Mark Winey which referenced a paper by Byers (Byers, B. and Goetsch, L. (1991) Preparation of yeast cells for thin section electron microscopy. Methods Enzymol. 194: 602-608.) This procedure involves spheroplasting which removes the cell wall, thus allowing good infiltration. The cells turned out looking great but the people I did the work for wanted to see more membranes.
Another good reference for fixing yeast is a paper by Robin Wright (Wright,R. (2000) Transmission Electron Microscopy of Yeast. Microsc. Res. Tech. 51:496-510). She gives alot of helpful hints for fixing yeast. She gives a detailed procedure in the paper using potassium permanganate as a post fixative. If you use potassium permanganate as a post fixative you don't have to spheroplast the cells (according to the paper, I have not tried it yet). Of course we all know that potassium permanganate extracts much of the cytoplasm, which is perfect for viewing membranes. I hope this will be helpful.
Janice Pennington Senior Electron Microscopist Nephrology/Anatomy and Cell Biology Indiana University School of Medicine 635 Barnhill Drive, MS 5065 Indianapolis, IN 46202 Phone (317) 274-8730 Fax (317) 278-2040
2. ----------------- Hello, All-
Last year a couple of students from BYU Hawaii were interested in looking for copper in the mitochondria of yeast by EELS and/or ESI, using our LEO 912 EFTEM. The first (but by no means the only) stumbling block was getting decent ultrastructure, or at least good enough that they could identify the mitochondria, and good enough infiltration that 25-30 nm sections could be obtained.
I do not have any experience with yeast and so I am open to any and all hints, tips, and suggestions!
If this works out, I invite you all to come to see the results at M&M 2005 in Honolulu next August.
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* ****************************************************************************
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 4 12:28:39 2004
The Low Angle Ion Thinner you are referring to is the IV3 from Technoorg-Linda in Budapest. Actually, they make the IV3 with high energy guns, low energy guns as well as a focussed ion gun. They also produce the Gentle Mill which is a dedicated low energy ion polisher.
As the distributor for Technoorg-Linda products in North America, we do offer the carbon/epoxy product that you reference. I will contact you offline with the part number and pricing.
DISCLAIMER: South Bay Technology produces equipment and supplies as described above and, therefore, has a vested interest in promoting their use.
Best regards-
David
hkonishi-at-indiana.edu wrote:
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From MicroscopyL-request-at-ns.microscopy.com Mon Oct 4 13:25:49 2004
Hi Tina, Yeast cells freeze quite nicely. I think the closest high pressure freezer to you is located in Kent McDonald's lab at Berkeley .
Beth
On Thursday, September 30, 2004, at 05:53 PM, Tina Carvalho wrote:
} } } ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } Hello, All- } } Last year a couple of students from BYU Hawaii were interested in } looking } for copper in the mitochondria of yeast by EELS and/or ESI, using our } LEO } 912 EFTEM. The first (but by no means the only) stumbling block was } getting decent ultrastructure, or at least good enough that they could } identify the mitochondria, and good enough infiltration that 25-30 nm } sections could be obtained. } } I do not have any experience with yeast and so I am open to any and all } hints, tips, and suggestions! } } If this works out, I invite you all to come to see the results at M&M } 2005 } in Honolulu next August. } } 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* } *********************************************************************** } ***** } } } ********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) *******************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
But how does one know which marker pens are OK and which are not?
cheers
rtch
} } One thing of note.... } } DO NOT use sharpie markers or any other acid base pen to label cd's or } dvd's. The acid in these pens etches through the top of the disk } destroying your data. } } Dave Crone B.E. (Mechanical) } Engineer-in-Training } Department Assistant } Metallurgical Lab } Mechanical Engineering } College of Engineering } University of Saskatchewan } 57 Campus Drive } Saskatoon, SK } S7N 5A9 } Phone: (306) 966-5461 } Fax: (306) 966-5427
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 4 18:24:47 2004
It is a safe bet that if they are sold on the premise that they are cd safe they should be good
Dave Crone B.E. (Mechanical) Engineer-in-Training Department Assistant Metallurgical Lab Mechanical Engineering College of Engineering University of Saskatchewan 57 Campus Drive Saskatoon, SK S7N 5A9 Phone: (306) 966-5461 Fax: (306) 966-5427 E-mail: dgc132-at-mail.usask.ca dgc132-at-gmail.com -----Original Message----- } From: Ritchie Sims [mailto:r.sims-at-auckland.ac.nz] Sent: Monday, October 04, 2004 3:06 PM To: Dave Crone; 'MICROSCOPY'
But how does one know which marker pens are OK and which are not?
cheers
rtch
} } One thing of note.... } } DO NOT use sharpie markers or any other acid base pen to label cd's or } dvd's. The acid in these pens etches through the top of the disk } destroying your data. } } Dave Crone B.E. (Mechanical) } Engineer-in-Training } Department Assistant } Metallurgical Lab } Mechanical Engineering } College of Engineering } University of Saskatchewan } 57 Campus Drive } Saskatoon, SK } S7N 5A9 } Phone: (306) 966-5461 } Fax: (306) 966-5427
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 4 19:10:31 2004
I have used Sharpies many times to mark disks, and have never seen a problem yet. I should add that I always use top-coated disks.
John Mardinly Intel
-----Original Message----- } From: Ritchie Sims [mailto:r.sims-at-auckland.ac.nz] Sent: Monday, October 04, 2004 2:06 PM To: Dave Crone; 'MICROSCOPY'
But how does one know which marker pens are OK and which are not?
cheers
rtch
} } One thing of note.... } } DO NOT use sharpie markers or any other acid base pen to label cd's or } dvd's. The acid in these pens etches through the top of the disk } destroying your data. } } Dave Crone B.E. (Mechanical) } Engineer-in-Training } Department Assistant } Metallurgical Lab } Mechanical Engineering } College of Engineering } University of Saskatchewan } 57 Campus Drive } Saskatoon, SK } S7N 5A9 } Phone: (306) 966-5461 } Fax: (306) 966-5427
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 4 19:36:57 2004
I agree with Henk and Mike. I believe CDRs ARE archival grade if handled properly. Even those thousand year old clay tablets, suggested in jest, are almost useless if broken or turned to dust from improper handling.
I did not read the whole article but I what to address the Sharpie® acidic ink problem. In the interest of having all this CDR stuff in one posting, I offer this web information:
I used Sharpie pens for years on CDRs without a problem. The acidic pen posting raised these questions: Were my Sharpie® pens the acidic types? Was my first official US flag research CD going to disintegrate? So, I looked to SandfordCorp.com to see what they said about acidic pens.
Quoting: "There are two criteria, which identify "Acid Free" Products: 1. The product must contain no added acid 2. The product must contain an ink with a pH specification of 7 or above, or the product has no measurable pH because it contains a solvent - based ink or it is a solid. (The pH cannot be measured if the ink does not contain water)
Product Color (snip other types of markers that are not permanent)
Permanent Markers 7000 Marker All Colors 7007 Marker All Colors Deluxe Marker - All Colors Not Black King Size Marker - All Colors Not Black Liquid Tip Marker All Colors Magnum 44 - All Colors Not Black Mean Streak All Colors Rub-A-Dub Black Sharpie Autograph Black Label Pen Black Penguin Freezer Wrap Marker Black T.E.C. Markers Black (Trace Element free pens) Sharpie Industrial Black " End quote.
It should be obvious what to use. I used black Industrial and Label pens without any problems, IMO.
Disclaimer: I don't work for Sanford Corp but I like Sharpie® pens.
My thanks to Hendrik O. Colijn at The Ohio State University for providing the original reference.
Paul Beauregard Senior Research Associate
----------------------------------------------------------------------- } Original Sharpie Pen post: } One thing of note.... } DO NOT use sharpie markers or any other acid base pen to label cd's or } dvd's. The acid in these pens etches through the top of the disk destroying } your data. } } Dave Crone B.E. (Mechanical) } End Beauregard postings. ------------------------------------------- At 10:28 AM 9/27/04 -0700, Michael O'Keefe wrote: } } Henk, } } That's an extremely informative and useful article. Thank you! } } It was interesting to learn that CD RW and DVD RW (bits encoded in metal structure } by a phase change from crystalline to amorphous) are less stable over time than CD } R and DVD R (bits encoded in degradable organic dyes). And that RW discs that are } "cycled" a lot (many re-writes) will degrade faster. } } The best advice seems to be to use R (write once) discs (CD or DVD) and store them } vertically in a cool, low-light environment. Of course, transferring to fresh } media every year or so couldn't hurt... } } Mike } } "Hendrik O. Colijn" wrote: } } } ------------------------------------------------------------------------------ } } } } Hi all, } } } } I just ran across this reference in our local newspaper about the care and } } storage of CDs and DVDs. It is copublished by CLIR (Council on Library and } } Information Resources) and NIST, so it should be reliable info. } } } } http://www.clir.org/pubs/reports/pub121/contents.html } } } } From my quick run through of the article, it appears that DVDs may be } } inherently more stable than CDs. } } } } Cheers, } } Henk } } } } Hendrik O. Colijn colijn.1-at-osu.edu } } Campus Electron Optics Facility Ohio State University } } (614) 292-0674 http://www.ceof.ohio-state.edu } } Time is that quality of nature which keeps events from happening all at } } once. Lately it doesn't seem to be working.
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 4 19:58:26 2004
According to the article I read, the issue seems to be damage to the lacquer on the top surface of the CD. The author of the article recommends water based markers or perhaps alcohol based, but not other solvent based. For example, I've got a marker which appears to be xylene-based (sniff test!) and probably damaging to the protective lacquer. I certainly found it interesting that the top-surface is more delicate than the bottom!
For details, see below... http://www.clir.org/pubs/reports/pub121/contents.html
Cheers, Henk
At 05:06 PM 10/4/2004, Ritchie Sims wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hendrik O. Colijn colijn.1-at-osu.edu OSU Campus Electron Optics Facility www.ceof.ohio-state.edu 040 Fontana Labs, 116 W. 19th Ave
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 4 21:07:25 2004
Optical Elements Corporation (OPELCO) has current openings for an Image Analysis Sales Specialist and a Confocal Microscopy Sales Specialist in the Mid-Atlantic region. If interested, please click on the following link or contact me directly.
http://www.opelco.com/employmentcontact.htm
Best Regards,
Cliff Glier COO OPELCO 105 Executive Drive Suite 100 Dulles, VA 20166 703.471.0080 x230 703.904.9432 (fax) cglier-at-opelco.com www.opelco.com
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 5 10:29:47 2004
Thanks for your info. I went to the Sanford site and noticed that they provide MSDS sheets for their markers. Interestingly, Sharpies do not all use the same solvents. I'm not an organic chemist, so I would appreciate if someone would correct me... If I remember correctly, n-propanol and n-butanol are alcohols and should not affect the lacquer. I don't know about diacetone alcohol or about the ethers. Based on my uneducated guess, I would probably avoid the naptha solvents in the "Sharpie Professional" series.
Can anyone provide additional info about lacquers and solvents?
Sharpie Fine Point Permanent, Sharpie Twin-tip, Super Sharpie, Super Sharpie Twin-tip n-propanol, n-butanol, diacetone alcohol
Sharpie Ultra Fine Point n-propanol, n-butanol, diacetone alcohol, isopropyl alcohol, ethyl alcohol
Sharpie Industrial Fine Point ethylene glycol monobutyl ether, ethylene glycol monoethyl ether, ethyl alcohol
Sharpie Professional nitroparaffin solvent, solvent naptha
Cheers, Henk Colijn
At 08:41 PM 10/04/04, Beauregard wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://www.ceof.ohio-state.edu Time is that quality of nature which keeps events from happening all at once. Lately it doesn't seem to be working.
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 5 12:41:49 2004
I've recently started taking birefringence photographs of Congo Red stained tissue. I've noticed that over time, the very same field doesn't appear the same, and that the labeled surface varies up to 25%.
Everything being very stable (all screws firmly screwed, polarizer glued to its base), I'm wondering if anyone has performed a test to check the warm-up period of a halogen 6 volt lamp. The company who sells the lamp says 5 to 10 minutes, but could it be longer?
Thank you!
Marie-Claude Belanger
_________________________________________________________________ Des mécanismes de contrôle parental puissants permettent à votre enfant de découvrir tout ce qu’Internet a à offrir. http://join.msn.com/?pgmarket=fr-ca&page=features/parental&ST=1&xAPID=1983&DI=2043 Commencez dès maintenant à profiter de tous les avantages de MSN Premium et obtenez les deux premiers mois GRATUITS*.
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 5 12:57:22 2004
Does anyone have a good reciepe for TEM fixation of ebryonic cell cultures? (ie 293 kidney). Are these cell's osmolarities generally higher or lower than other cell types. Thanks.
Mike Delannoy
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 5 16:38:06 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (monica.iliescu-at-polymtl.ca) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, October 5, 2004 at 10:47:16 ---------------------------------------------------------------------------
Question: Does some know how prepare the biological samples for ESEM observations? We have an Quanta FEG 200, FEI Company, and we want to prepare for observation a sample from chitosan solution, to see the chitosan nanoparticles. Thank you!
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (aiqbal-at-email.arizona.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, October 5, 2004 at 13:21:33 ---------------------------------------------------------------------------
Question: Please if some one could help me with this. I would like to know if there is any software available to measure thickness and composition of the sample using SEM/EDS. Where can I get it from?
I am looking for help imaging active drug particles in an aqueous pharmaceutical suspension. I am trying to locate someone who can view frozen sections of this material in the 100 angstrom size range. I believe this will require an environmental SEM with a cryogenic stage. I would appreciate any help. Thank you.
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 5 17:05:09 2004
Hi all, I have not used NIH Image or Scion Image and a student in my department has the following question:
} I'm looking at leaf morphology in a number of tree species, and I've } got 100's of flatbed leaf scans that I'm trying to analyze using Scion } Image (a PC spin-off of NIH Image) and ImageJ. The problem I'm having } is that the program is recognizing (and measuring) the background as } well as the leaf in its area calculations. Do you have any advice or } could you help in any way in dealing with this problem?
Any help would be greatly appreciated! thanks in advance, Beth
********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) *******************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
} } I believe CDRs ARE archival grade if handled } properly.
That (handled properly) is indeed the problem with this class of media.
As was mentioned in the report referenced by an earlier post, (if you read between the lines), this media CAN be very easily damaged.
As an IT Manager, I would suggest you think about what you mean by "Archive" class media.
If you mean something that you can hide away in a controlled environment, and carefully access at some point in the future, then maybe this class of media is appropriate.
If you mean something that you can use for routine "archive" access (as opposed to "on-line" access) I would strongly suggest that the media CAN be too sensitive to physical damage (more so on the label side than the "bottom" side).
If you mean something that will absolutely, positively be there when you need it for "disaster recovery", I would suggest that this isn't the media you want for that.
I don't mean to say the media isn't durable, or won't last. Just that there is a chance that a single disk may be easily damaged, either by natural causes (thermal cycling, humidity, etc.) or accidentally by routine handling. My friend Murphy says that this will happen on the most important area of the most important disk at the most inopportune time.
I have lived the nightmare of backup systems not working for a number of reasons. You don't want to go there.
John W. Raffensperger, Jr. IT Manager Apache Stainless Equipment
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 5 17:35:18 2004
I would like to know the best way to prepare ion-thinning samples of small particles. The size is several 10 um. I know that FIB or microtome might work better, but I want to try ion thinning. The material is mineral like SiO2.
If you embed small grains in epoxy, the grains drop during milling in many cases because thinning rates of sample and epoxy are different. Also, rough surface that are produced by preferential etching is another problem. It is hard to make good ion-thinning sample of small grains in general.
Using Araldite (AT1) containing C or M-bond 610 can reduces the problem of preferential etching of glue (Barna 1992, Westman et at 1999, McCaffrey and Barna 1997). (SBT sells "Carbon powder & Araldite-Type AT1" for low angle ion milling). Photo-resist is "hard" in ion milling (Yoshioka at al 1995).
I do not have any other idea that may reduce the problem in preferential etching. If you have any suggetions, please advise. Also, if someone used AT1 with Gatan Dual Mill or PIPS, I would like to know if you got better result or not.
Thank you,
Hiromi Konishi, Ph.D. Indiana University www.unm.edu/~hkonishi
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 5 19:22:37 2004
.. does the student threshold and binarize the picture before to do the analysis ?
1) Threshold and choose the level he wants
2) Binarize (black & white picture, 0 or 256)
3) Analyse (and if the program take everything & not only the particles, try invert picture. I don't remember if NIH or Scion Image consider the white or the black spots as particles...).
Hope it helps
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Phone: 001-617-667-2143 Fax: 001-617-667-8210
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 6 00:37:19 2004
Friends on Darwin, a new italian journal on science and science policy, there is an article on Multiphoton microscopy "Il radioso futuro della microscopia ottica" and also the cover has been dedicated to multiphoton microscopy. I'd like to thank Kenneth Dunn for providing one of the images of the article collected and rendered by Ruben Sandoval at the Indiana Center for Biological Microscopy. Website is www.darwinweb.it. All my best ALby
------------------------------------------------------------------------ --------------------------- Alberto Diaspro, Department of Physics, University of Genoa Via Dodecaneso 33, 16146 Genova, Italy facsimile +39-010314218 - voice +39-0103536426/480/309 URL: http://www.lambs.it http://www.wiley.com/WileyCDA/WileyTitle/productCd-0471409200.html ------------------------------------------------------------------------ --------------------------
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 6 03:05:53 2004
layer thickness measurement within the Electron Microscope and with the EDX-spectrometer with electron excitation is possible, but is faced to a lot of limits. The penetration-range of electrons in specimen is strongly limited. If the layer is going to become thicker (depends from primary electron energy, typically 1 micron or less), saturation is reached, no reliable results are possible. The calculation of all interactions are complex. If there are multi-layers, the electrons have to travel through the top layers... Therefore layer-measurements are only possible with very thin layers.
With an X-ray excitation the entire situation is more friendly and therefore more reliable. Thicknesses of 1 nm up to 50 microns are possible to deal with. Because of the larger range of X-ray penetration in specimen, multi-layers are not a problem. An X-ray excitation in SEM is available now and proper software for calculation thickness and concentrations with X-ray excitation, as well: see: http://www.ifg-adlershof.de/sem.htm
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (aiqbal-at-email.arizona.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, October 5, 2004 at 13:21:33 ---------------------------------------------------------------------------
Question: Please if some one could help me with this. I would like to know if there is any software available to measure thickness and composition of the sample using SEM/EDS. Where can I get it from?
On Tue, 2004-10-05 at 15:23, Chiphead wrote: } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } } } I believe CDRs ARE archival grade if handled } } properly. } } That (handled properly) is indeed the problem with this class of media. } } As was mentioned in the report referenced by an earlier post, (if you } read between the lines), this media CAN be very easily damaged. } } As an IT Manager, I would suggest you think about what you mean by } "Archive" class media. } } If you mean something that you can hide away in a controlled } environment, and carefully access at some point in the future, then } maybe this class of media is appropriate. } } If you mean something that you can use for routine "archive" access (as } opposed to "on-line" access) I would strongly suggest that the media CAN } be too sensitive to physical damage (more so on the label side than the } "bottom" side). } } If you mean something that will absolutely, positively be there when you } need it for "disaster recovery", I would suggest that this isn't the } media you want for that. } } I don't mean to say the media isn't durable, or won't last. Just that } there is a chance that a single disk may be easily damaged, either by } natural causes (thermal cycling, humidity, etc.) or accidentally by } routine handling. My friend Murphy says that this will happen on the } most important area of the most important disk at the most inopportune } time. } } I have lived the nightmare of backup systems not working for a number of } reasons. You don't want to go there. } } John W. Raffensperger, Jr. } IT Manager } Apache Stainless Equipment } } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 6 08:43:55 2004
Already a couple of years ago, former colleagues did automated calcium-ratio measurements with CCD-camera's on individual (cardio)myocytes in cell cultures. They used a modified CCD-camera which allowed them to capture an image of an individual (cardio)myocyte every 10 milliseconds to monitor intracellular calcium.
I am wondering which type of camera could now be used to give the same time-resolution (10 msec. frame time) for individual cells (the hardware they used is now no longer available) ?
Reference:
Cornelissen, F., Wouters, L., Ver Donck, L., Verellen, G., Geerts, H. Automatic quantification of the effect of cardioprotective drugs in isolated myocytes. Bioimaging, 1993, 1, pp. 197-206.
Regards,
Peter Van Osta
---------------------------------------------- Peter Van Osta
In my opinion (for what it may be worth) tape has proven to be, and remains, the best "archival" media. It has proven to be the most durable, and most likely to be available when needed.
Raid would come into play not so much for archive, but for "availability". If you can live with the data being unavailable for a period of time while a new disk drive is put in place (if required) and the backup restored, then raid is not necessarily required. If you NEED to have the data available NOW, then raid provides some fault tolerance, and the ability to keep the data available while the recovery is in process.
Tape is not perfect, it can be destroyed, but it is less likely to have the types of single use failure that CD/DVD media are susceptible to. They can take an awful lot of environmental and physical abuse, and still be readable.
Don't get me wrong, CD/DVD have their place. I use them all the time. Just not for archive, and I never use them to hold the only copy of something (even on multiple disks) that needs to be kept. For cases where the data could not be kept on the hard drives, I would make a tape backup, then a set of CD/DVDs for the users.
John Raffensperger
--- Robb Westby {robbw-at-mxim.com} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } John, } Then what would a "good" method of data archive? } Hardware RAID and tape? } } --- } Robb Westby } Senior SEM/FIB Technician } Maxim Integrated Products } }
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 6 15:14:26 2004
Hello, Would using hepes as a buffer for reducing glutaraldehyde with Nh4Cl be a problem? I know hepes isn't compatible with osmium, but what about other IEM chemicals say tannic acid? thanks Mike D.
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 6 17:35:15 2004
We use HEPES for paraformaldehyde/glutaraldehyde in the primary fix, with 50 mM glycine in the blocking step, and with 1% osmium. why do you think it is incompatible with osmium?
At 03:18 PM 10/06/04, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (bwareham-at-utah.gov) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, October 6, 2004 at 16:06:52 ---------------------------------------------------------------------------
Question: Hi all, I'm looking for a protocol to prepare some gel samples for SEM. They are referred to in papers as hydrogels or organogels. Any help with this is greatly appreciated.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (njt3-at-u.washington.edu) from http://msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, October 6, 2004 at 15:14:41 ---------------------------------------------------------------------------
Email: njt3-at-u.washington.edu Name: Jeanie Taylor
Organization: Univ of Wash
Education: Graduate College
Location: Seattle, Wa
Question: I am trying to calibrate the scale on an Olympus BH-2 compound scope. How do I check the measurement of the sacle in the eyepiece?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mahtab977-at-yahoo.com) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, October 6, 2004 at 16:48:16 ---------------------------------------------------------------------------
Question: I was wondering if anyone has had any experience using Si-chips (from Ted Pella) for growing cells. My purpose is to visualize attachment appendages of gram-negative bacteria, so a smooth surface is key. I'm not sure how much better Si-chips are than glass coverslips, and I also don't know how how to sterilize them (autoclave or ethanol?).
Furthermore, I want to thank you all for your responses to a past question of mine about HMDS drying- the procedure/tips I recieved were very useful.
On Oct 6, 2004, at 4:22 PM, by way of MicroscopyListserver wrote:
} I'm looking for a protocol to prepare some gel samples for SEM. They } are referred to in papers as hydrogels or organogels. Any help with } this is greatly appreciated. } Dear Beverly, Robert Apkarian has worked with these substances a lot, so check his recent papers (if you have not already done so). Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 6 21:14:30 2004
Another media which may be considered "archival quality" is magneto-optical (MO) disks. They are mounted in the case and have a shelf life expectation of 100+ years. As far as I know most of the government agencies used this media to store our records at FBI and INS. MOs may be destroyed by physical force (hammer?) or extreme temperature (fire). Tapes are great too and more practical than MOs. The greatest disadvantage of MOs is that they are slow and capacity is not very impressive (a couple of Gigs, which is nothing nowadays). Modern CD/DVDs may not be considered as a good storage solution. Meantime, we are using them in everyday life (with some risk). The best known to me storage media is ceramic tablets. Sergey
At 08:12 AM 10/6/2004, 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 10833 Le Conte Ave, Room 33-080 Los Angeles, CA 90095
Bioptechs does not make a habit of this type of announcement in this forum. However, in this case I, think it is appropriate.
For the past 12 years we have been asked , by many users of this list, for a micro-environmental cell culture chamber system for upright microscopes. I am pleased to announce that we have finally completed it! It is called the FCS3. It is based on the popular FCS2 technology. This system will enable any upright microscope equipped with a digital camera to acquire long term ,live-cell , time-lapse images of living organisms including mammalian specimens. Now, those of you without an inverted microscope can easily accommodate live-cell experiments with the same ease as inverted scope users. Ideal for multi user facilities! Additional information is available at: http://www.bioptechs.com/Products/FCS3/fcs3.html (This link will take you directly to the appropriate page)
If you are offended by this posting please respond directly to me with your admonishment.
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 7 00:30:19 2004
I have been following the tread since we also store data. (as do all EM units!) I did not see the mentioning of hardisks as a option. I have thrown away (beginning of last year) my vintage 20meg hard disk that was purchased in 1989. My Pentium 1 was scrapped. It was still working perfectly with all the data intact. Hot-swapping allows fast and reliable backup easy. The cost of HD has dropped dramatically. Re installing is fast (hot swapping) Just a thought.
-----Original Message----- } From: Sergey Ryazantsev [mailto:sryazant-at-ucla.edu] Sent: Thursday, October 07, 2004 4:19 AM To: Microscopy-at-microscopy.com
Another media which may be considered "archival quality" is magneto-optical (MO) disks. They are mounted in the case and have a shelf life expectation of 100+ years. As far as I know most of the government agencies used this media to store our records at FBI and INS. MOs may be destroyed by physical force (hammer?) or extreme temperature (fire). Tapes are great too and more practical than MOs. The greatest disadvantage of MOs is that they are slow and capacity is not very impressive (a couple of Gigs, which is nothing nowadays). Modern CD/DVDs may not be considered as a good storage solution. Meantime, we are using them in everyday life (with some risk). The best known to me storage media is ceramic tablets. Sergey
At 08:12 AM 10/6/2004, 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 10833 Le Conte Ave, Room 33-080 Los Angeles, CA 90095
We had a quick look at hydogels by putting them wet in an ESEM.
Dave
On Wed, 06 Oct 2004 18:22:35 -0500 by way of MicroscopyListserver {bwareham-at-utah.gov} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } 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 (bwareham-at-utah.gov) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, October 6, 2004 at 16:06:52 } --------------------------------------------------------------------------- } } Email: bwareham-at-utah.gov } Name: Beverly Wareham } } Organization: Utah Veterinary Diagnostic Laboratory } } Title-Subject: [Microscopy] [Filtered] hydrogels/organogels } } Question: Hi all, } I'm looking for a protocol to prepare some gel samples for SEM. They are referred to in papers as hydrogels or organogels. Any help with this is greatly appreciated. } } Thanks, } Beverly Wareham } Utah Veterinary Diagnostic Laboratory } Logan, Utah } } --------------------------------------------------------------------------- } } } } This incoming email to UWE has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
This email has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 7 08:00:02 2004
Mahtab Shahkarami wrote: ============================================================================ = Question: I was wondering if anyone has had any experience using Si-chips (from Ted Pella) for growing cells. My purpose is to visualize attachment appendages of gram-negative bacteria, so a smooth surface is key. I'm not sure how much better Si-chips are than glass coverslips, and I also don't know how how to sterilize them (autoclave or ethanol?). ============================================================================ I have been lead to believe that at least some are using our own gold coated Si-Chips for the growing of cells, see URL http://www.2spi.com/catalog/submat/gold-coated-silicon-chips.shtml
The description of the uncoated silicon chips, also used for cell growth, is given on URL http://www.2spi.com/catalog/standards/silicon-chip-sample.shtml The smoothness of the silicon surface is comparable to that of a glass cover slip. The silicon used for their production is standard electronics industry grade silicon wafers.
The uncoated silicon chips can certainly be autoclaved, however, we don't have information on the survivability of the gold coated substrates under autoclave conditions. We would appreciate knowing if anyone has such information since we are asked that question from time to time.
Disclaimer: SPI Supplies manufactures both uncoated and gold coated silicon chip substrates for cell growth applications.
Chuck =================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 7 08:51:01 2004
My company is considering obtaining an Hitachi 806 cold field emitter SEM. The organization that if offering it has not been pleased with its performance, but is offering a really good deal. I was wondering if anyone has experience with this tool? If so I would appreciate your comments.
Thanks,
Gerhard -- Gerhard S. Schoenthal, PhD Director of Laboratories Virginia Diodes, Inc. 321 West Main Street Charlottesville, VA 22903
W: 434.297.3257 M: 434.409.7760 F: 530.884.5710
W: Schoenthal-at-vadiodes.com
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 7 08:59:21 2004
My company is considering obtaining an Hitachi 806 cold field emitter SEM. The organization that if offering it has not been pleased with its performance, but is offering a really good deal. I was wondering if anyone has experience with this tool? If so I would appreciate your comments.
Thanks,
Gerhard -- Gerhard S. Schoenthal, PhD Director of Laboratories Virginia Diodes, Inc. 321 West Main Street Charlottesville, VA 22903
W: 434.297.3257 M: 434.409.7760 F: 530.884.5710
W: Schoenthal-at-vadiodes.com
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 7 09:10:02 2004
Tom, I once read on this listserver that organic buffers shouldn't be used with osmium because they could react (oxidized?). Although I will admit that I have used reduced osmium with hepes with no noticeable artifacts (maybe due to the lower oxidation state). I want to know if Hepes can be used as the only buffer in a post-embed IEM protocol up to uranyl acetate.
thanks Mike Delannoy
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 7 09:39:09 2004
To answer your question about gold surviving autoclave on Si substrate, you must state what the base metal is, e.g. Ti, Ti-Pt? I don't think you put Au directly on the wafer without a base metal.
Peter Tomic Agere Systems Allentown, PA
-----Original Message----- } From: Garber, Charles A. [mailto:cgarber-at-2spi.com] Sent: Thursday, October 07, 2004 10:04 AM To: MICROSCOPY BB
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Mahtab Shahkarami wrote: ======================================================================== ==== = Question: I was wondering if anyone has had any experience using Si-chips (from Ted Pella) for growing cells. My purpose is to visualize attachment appendages of gram-negative bacteria, so a smooth surface is key. I'm not sure how much better Si-chips are than glass coverslips, and I also don't know how how to sterilize them (autoclave or ethanol?).
======================================================================== ==== I have been lead to believe that at least some are using our own gold coated Si-Chips for the growing of cells, see URL http://www.2spi.com/catalog/submat/gold-coated-silicon-chips.shtml
The description of the uncoated silicon chips, also used for cell growth, is given on URL http://www.2spi.com/catalog/standards/silicon-chip-sample.shtml The smoothness of the silicon surface is comparable to that of a glass cover slip. The silicon used for their production is standard electronics industry grade silicon wafers.
The uncoated silicon chips can certainly be autoclaved, however, we don't have information on the survivability of the gold coated substrates under autoclave conditions. We would appreciate knowing if anyone has such information since we are asked that question from time to time.
Disclaimer: SPI Supplies manufactures both uncoated and gold coated silicon chip substrates for cell growth applications.
Chuck =================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 7 09:44:27 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (schoenthal-at-vadiodes.com) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, October 7, 2004 at 09:03:40 ---------------------------------------------------------------------------
Email: schoenthal-at-vadiodes.com Name: Gerhard Schoenthal
Organization: Virginia Diodes, Inc.
Title-Subject: [Microscopy] [Filtered] Opinions on Hitachi 806 SEM
Question: Hi,
My company is considering obtaining an Hitachi 806 cold field emitter SEM. The organization that if offering it has not been pleased with its performance, but is offering a really good deal. I was wondering if anyone has experience with this tool? If so I would appreciate your comments.
Thanks,
Gerhard -- Gerhard S. Schoenthal, PhD Director of Laboratories Virginia Diodes, Inc. 321 West Main Street Charlottesville, VA 22903
I have seen HEPES react with KMnO4 (it literally turns the mix into a gel that then breaks down into time into a precipitate). Never had a problem with either 1% osmium or 1% reduced osmium. After osmium, we usually rinse with dH2O before ethanol dehydration since by this time the tissue is no longer osmotically sensitive. We have done uranyl acetate in ethanol, or sodium acetate buffer, or water at this stage. I haven't seen a lot of difference but I think others are strong advocates of uranyl in the ethanol or sodium acetate. If by post-embedding IEM you mean doing immunostaining of osmicated, embedded tissues, you have a low probability of success with most antigens in my experience. I have had a few antigens survive osmication but generally only ones in high abundance to start. Most immuno EM work using staining of sections would use aldehyde fixation (formaldehyde only, or with low glutaraldehyde ~0.2% or sometimes a standard 2% PF + 2.5% glut) followed by buffer rinses and then maybe a uranyl acetate en bloc staining and dehydration before embedding in an immuno friendly plastic like K4M, HM20, LRW, or LRG. LR Gold is my current favorite but we find the choice is dependent on the antigen and we need to re-invent the wheel each time and try them all. During our immunostaining of sections, we use a HEPES buffer (70 mM NaCl, 30 mM HEPES, 2 mM CaCl2, pH 7.4) for the antibodies (usually with 0.1% BSA). good luck.
At 09:12 AM 10/07/04, you wrote: } Tom, } I once read on this listserver that organic } buffers shouldn't be used with osmium because they } could react (oxidized?). Although I will admit that } I have used reduced osmium with hepes with no noticeable artifacts (maybe } due to the lower oxidation } state). I want to know if Hepes can be used as the } only buffer in a post-embed IEM protocol up to } uranyl acetate. } } thanks } Mike Delannoy
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
} Date: Thu, 07 Oct 2004 09:58:33 -0500 } To: MICHAEL J DELANNOY {delannoy-at-jhmi.edu} } From: Tom Phillips {phillipst-at-missouri.edu} } Subject: [Microscopy] Re: Re: re: Hepes and reduction } Cc: microscopy } } I have seen HEPES react with KMnO4 (it literally turns the mix into a gel } that then breaks down into time into a precipitate). Never had a problem } with either 1% osmium or 1% reduced osmium. After osmium, we usually } rinse with dH2O before ethanol dehydration since by this time the tissue } is no longer osmotically sensitive. We have done uranyl acetate in } ethanol, or sodium acetate buffer, or water at this stage. I haven't seen } a lot of difference but I think others are strong advocates of uranyl in } the ethanol or sodium acetate. If by post-embedding IEM you mean doing } immunostaining of osmicated, embedded tissues, you have a low probability } of success with most antigens in my experience. I have had a few antigens } survive osmication but generally only ones in high abundance to } start. Most immuno EM work using staining of sections would use aldehyde } fixation (formaldehyde only, or with low glutaraldehyde ~0.2% or sometimes } a standard 2% PF + 2.5% glut) followed by buffer rinses and then maybe a } uranyl acetate en bloc staining and dehydration before embedding in an } immuno friendly plastic like K4M, HM20, LRW, or LRG. LR Gold is my } current favorite but we find the choice is dependent on the antigen and we } need to re-invent the wheel each time and try them all. During our } immunostaining of sections, we use a HEPES buffer (70 mM NaCl, 30 mM } HEPES, 2 mM CaCl2, pH 7.4) for the antibodies (usually with 0.1% } BSA). good luck. } } } } At 09:12 AM 10/07/04, you wrote: } } Tom, } } I once read on this listserver that organic } } buffers shouldn't be used with osmium because they } } could react (oxidized?). Although I will admit that } } I have used reduced osmium with hepes with no noticeable artifacts (maybe } } due to the lower oxidation } } state). I want to know if Hepes can be used as the } } only buffer in a post-embed IEM protocol up to } } uranyl acetate. } } } } thanks } } Mike Delannoy } } Thomas E. Phillips, PhD } Professor of Biological Sciences } Director, Molecular Cytology Core } 3 Tucker Hall } University of Missouri } Columbia, MO 65211-7400 } } 573-882-4712 (office) } 573-882-0123 (fax) } PhillipsT-at-missouri.edu
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Silicon wafers behave pretty much like glass. They are ultra-smooth, cells can be grown on them without prior coating with gold, and they are rsistant to most reagents you are likely to use. An advantage of silicon as a substrate for SEM of cell monlayers is that the substrate has some conductivity, the silicon background charges rather less than glass for a given accelerating voltage and beam current, and therefore thinner sputtered coatings of gold/palladium, platinum or chromium can be employed. A disadvantage is they are more expensive than coverslips, and thicker. Chris
----- Original Message ----- } From: "Garber, Charles A." {cgarber-at-2spi.com} To: "MICROSCOPY BB" {Microscopy-at-MSA.Microscopy.com} Sent: Thursday, October 07, 2004 3:04 PM
Continuing the thread, Some have cited acid-based markers as a potential problem. Paul Beauregard offered a site at Sanford listing their acid-free markers. It was not clear whether Sharpie Fine Point markers were included or not, so I posed the question to their customer service department asking for clarification. I received the following two responses.
First, they stated that their markers are not acid-free. Next, they stated that their markers have been tested on CDs without problems.
I guess that simply means that no problem has been observed yet. Since it is a case of trying to prove a negative, I will go ahead and continue to use them unless someone can definitely show that the Fine Point markers have caused a problem. But I suppose I will still pick up a 'certified' CD marker the next time I go to the store.
Warren
} From: {consumer.service-at-sanfordcorp.com} } To: {wesaia-at-iastate.edu} } Subject: [Microscopy] Re: 000356732A, reply from Sanford, Paper Mate, Sharpie, } uni-ball, Eldon Office Solutions or Foohy.com web-site. } Date: Tue, 5 Oct 2004 19:00:25 -0400 } } Hello Warren, } } We received your recent inquiry regarding Sharpie® Fine Point Markers. } } The Sharpie® Fine Point Markers are not acid free. } } Thank you for e-mailing us and for your support of Sanford products. } } Sanford Consumer Affairs } } 000356732A
} From: {consumer.service-at-sanfordcorp.com} } To: {wesaia-at-iastate.edu} } Subject: [Microscopy] Re: 000356732B, reply from Sanford, Paper Mate, Sharpie, } uni-ball, Eldon Office Solutions or Foohy.com web-site. } Date: Thu, 7 Oct 2004 11:00:05 -0400 } } Hello Warren, } } Although the Sharpie Markers are not acid free, they have been found safe } for use on CD's and DVD’s by our lab. Our lab has conducted extensive } testing on these markers, and found since they are alcohol based, they are } safe for that application. } } Thank you for e-mailing us and for your support of Sanford products. } } Sanford Consumer Affairs } } 000356732B ------------- } Date: Mon, 04 Oct 2004 20:41:11 -0400 } To: Microscopy-at-microscopy.com } From: Beauregard {beaurega-at-westol.com} } Subject: [Microscopy] Re: Re: CD/DVD as archival media revisited } } Hi, } } I agree with Henk and Mike. I believe CDRs ARE archival grade if handled } properly. Even those thousand year old clay tablets, suggested in jest, } are almost useless if broken or turned to dust from improper handling. } } I did not read the whole article but I what to address the Sharpie® acidic } ink problem. In the interest of having all this CDR stuff in one posting, } I offer this web information: } } I used Sharpie pens for years on CDRs without a problem. } The acidic pen posting raised these questions: } Were my Sharpie® pens the acidic types? } Was my first official US flag research CD going to disintegrate? } So, I looked to SandfordCorp.com to see what they said about acidic pens. } } http://www.sanfordcorp.com/sanford/consumer/jhtml/help/sanford_help_922.jhtml } } Paul Beauregard } Senior Research Associate
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 46 Town Engineering Ames IA, 50011-3232
A colleague is interested in using a centrifuge to spin cells down on to glass slides. We would like the cells to be in multiple discrete spots on the slide. Anybody have tips on using this approach or places to buy such a device. Thanks, Tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
I have a question regarding the ultrastructural differences between Aspergillus and a gram positive bacteria. Does anyone out there know any differences that could be documented by the electron microscope? We have done all the special stains on this pathology specimen, but we are hoping the EM will help to further identify the organism we have in question.
Thanks for any help!
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
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 7 13:33:44 2004
Up to two 6-mm diameter cell spots per cytofunnel. I authored the user's' manual in 1982 or so.
Gary Gill
-----Original Message----- } From: Tom Phillips [mailto:phillipst-at-missouri.edu] Sent: Thursday, October 07, 2004 12:29 PM To: Microscopy-at-msa.microscopy.com
NESM Fall 2004 Meeting and Workshop on Cryo Techniques
The 2004/2005 meetings of the New England Society for Microscopy kick off this Fall with a pair of events on Cryo Techniques for specimen preparation. These techniques are useful for processing soft, high-volatile specimens for both SEM and TEM investigations. Soft, wet specimens are common in biological and human medical investigations. Perhaps less-appreciated are the relatively dry, soft polymers investigated by materials scientists and food technologists. The introductory event consists of three lectures on Cryo Techniques in Microscopy. It will be held on October 19th in the JEOL conference room in Peabody, Massachusetts. Registration starts at 5:30 PM, then a buffet dinner, and then the presentations. Those especially interested in learning Cryo skills should also plan to attend the two-day Workshop on Cryo-Ultramicrotomy on November 3-4. This workshop, sponsored or supported by NESM, RMC/Boeckeler, Harvard, and MIT, will be held at Harvard's Center for Imaging and Mesoscale Structures. The details of the meeting and workshop, including registration information, can be found on NESM's website http://prism.mit.edu:8083 under "current newsletter".
Registration forms/information for both events can be sent to: NESM Treasurer, Paul Bain c/o NESM, Harvard Medical School, Countway 212, 10 Shattuck Street, Boston, MA 02115 email: Paul_bain-at-hms.harvard.edu. The October 19th meeting costs $5 for members and $20 for non-members (you receive a one-year NESM membership) and the registration deadline is Friday, October 15th. The November 3-4 Workshop has a $50 professional registration and a $25 student registration. The registration deadline for the Workshop is Friday, October 22nd. (Register early for the workshop--space is limited!)
Peggy Sherwood Lab Associate, Photopathology Wellman Center for Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 7 16:40:44 2004
while you are aware of this, we should all remind ourselves that EM diagnostics of micro-organisms can be quite inexact. while negative stain of viruses will usually give good information concerning the family, even there we cannot go beyond the fact that the virus is a herpes, not a poxvirus, not that it is cytomegalovirus, or herpes zoster, or any other herpes, for that matter. we need the information of the infectious diseases and/or microbiology staff to be able to go from EM to specific diagnosis.
having said that, there are clear differences between the bacterial and fungal cells. most significant is the presence of internal membrane organelles with the eukaryotic cell, such as aspergillis, and the presence of the nucleoid in the prokaryotic cell. a second difference is that the fungal cell is significantly larger than the bacterial cell. by this, i mean 15-30microns as opposed to 1-5 microns. thirdly, the yeast and fungal cell walls and external structures are thicker (up to 500nm) than those seen with the gram positive bacteria. fourthly, with yeast, and some fungi, you will see budding scars. their orientation may give some idea as to the specific nature of the micro-organism you are seeing.
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-954 Cell:204-781-1502 Fax:204-789-3926
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 8 01:12:32 2004
We are looking at upgrading our facility here for photographing resected organs etc at the histopathology department. The new system will be based on digital cameras so that we can link the photos to the patient information and diagnosis in our database.
What I would like to know is what people are using these days in terms of macro lenses, ring flashes (if they exist for the newer cameras) and lighting systems, lamps, multiple swan-neck lighting.
Any tips would be appreciated from users and from manufacturers etc.
Gareth Morgan MPhil MSc FIBMS, Department of Laboratory Medicine (Labmed), Karolinska Institute, Karolinska University Hospital at Huddinge, F46 SE 141 86 Stockholm Sweden
OBS! Besöksadress: F-Huset, Forskningsgatan 2 F52, Rum 2.10. Laboratoriet för klinisk patologi och cytologi.
NB! Visiting address: Building F, Research Corridor 2 F52, Room 2.10. Clinical Histo- and Cytopathology Laboratory.
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 8 03:25:57 2004
Thank you all for sending information and suggestions about calcium ratio mesurements. The main issue was/is to find a camera-solution to achieve at least the the same framerate as was possible with the old hardware (now obsolete). Commonly used PAL video-cameras, have a framerate of about 25 frames per second (FPS, or 33 in NTSC countries) or lower. Popular digital cameras have advantages over "traditional" video-standard cameras, but not always achieve a framerate of 25/33 FPS.
What was done in the old system, was to put a "temporal" framesplitter on the camera, so it allowed to capture quarter frames at 1/4 of the frametime, which lead to a temporal sampling of 10 msec instead of 40 msec.
Chosing a modern camera which surpasses the old days solution is not a simple exercise. FireWire interfaces (800 Mbits/sec.) do not deliver the high bandwidth of CameraLink interfaces (base 2380 Mbit/sec. up to 7140 Mbit/sec.) and their performance goes down when using full-frame transfer. FireWire cameras are widely available, while the faster CameraLink cameras are not so popular. Every computer nowadays comes with a FireWire interface, but a CameraLink is not standard.
Already a couple of years ago, former colleagues did automated calcium-ratio measurements with CCD-camera's on individual (cardio)myocytes in cell cultures. They used a modified CCD-camera which allowed them to capture an image of an individual (cardio)myocyte every 10 milliseconds to monitor intracellular calcium.
I am wondering which type of camera could now be used to give the same time-resolution (10 msec. frame time) for individual cells (the hardware they used is now no longer available) ?
Reference:
Cornelissen, F., Wouters, L., Ver Donck, L., Verellen, G., Geerts, H. Automatic quantification of the effect of cardioprotective drugs in isolated myocytes. Bioimaging, 1993, 1, pp. 197-206.
Regards,
Peter Van Osta
---------------------------------------------- Peter Van Osta
Both Nikon and Olympus manufacture professional digital SLRs with top-quality macro lenses and flash systems. I always used to rate Olympus 35mm systems (based on OM2n, OM4ti) highly for their macro lenses, and TTL ringflash and macro-flash capability, and had always dreamed of the day when they would issue their updated system. Alas that was not to be, but an interesting new digital system has been launched by Olympus called E1 which uses the four thirds system. Macro lenses and a ringflash are available for this camera, I see, so this may turn into the digital equivalent of the OM system if it is equally well received in the market place.
Depending on how critical you are of issues like camera versatility, performance etc. you might get a long way towards a solution with a simpler camera like the Nikon Coolpix 4500. It has remarkable macro capability for the price, focussing to about 1 inch from front of lens, and there is a so-called ringflash accessory for it too, actually a ring of high-intensity white LEDs. I use one of these myself, and know many other scientists who swear by them for field-recording of specimens ranging from fluorescent gels to plants and fungi, even aurora borealis displays are within its capabilities.
I have nothing to gain commercially from these comments Hope this helps Chris
----- Original Message ----- } From: "Gareth Morgan" {Gareth.Morgan-at-labmed.ki.se} To: {Microscopy-at-msa.microscopy.com} Sent: Friday, October 08, 2004 7:17 AM
I don't know about gold coating, but titanium-coated silicon wafers are OK in the autoclave. Another way to sterilise them is to glow-discharge them.
Lesley Weston.
on 07/10/2004 7:04 AM, Garber, Charles A. at cgarber-at-2spi.com wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- --} - } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Mahtab Shahkarami wrote: } ============================================================================ } = } Question: I was wondering if anyone has had any experience using Si-chips } (from Ted Pella) for growing cells. My purpose is to visualize attachment } appendages of gram-negative bacteria, so a smooth surface is key. I'm not } sure how much better Si-chips are than glass coverslips, and I also don't } know how how to sterilize them (autoclave or ethanol?). } ============================================================================ } I have been lead to believe that at least some are using our own gold coated } Si-Chips for the growing of cells, see URL } http://www.2spi.com/catalog/submat/gold-coated-silicon-chips.shtml } } The description of the uncoated silicon chips, also used for cell growth, is } given on URL } http://www.2spi.com/catalog/standards/silicon-chip-sample.shtml } The smoothness of the silicon surface is comparable to that of a glass cover } slip. The silicon used for their production is standard electronics } industry grade silicon wafers. } } The uncoated silicon chips can certainly be autoclaved, however, we don't } have information on the survivability of the gold coated substrates under } autoclave conditions. We would appreciate knowing if anyone has such } information since we are asked that question from time to time. } } Disclaimer: SPI Supplies manufactures both uncoated and gold coated silicon } chip substrates for cell growth applications. } } Chuck } =================================================== } Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 } President 1-(800)-2424-SPI } SPI SUPPLIES FAX: 1-(610)-436-5755 } PO BOX 656 e-mail: cgarber-at-2spi.com } West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com } } } Look for us! } ############################ } WWW: http://www.2spi.com } ############################ } ================================================== } }
From MicroscopyL-request-at-ns.microscopy.com Sun Oct 10 15:42:36 2004
} } Depending on how critical you are of issues like camera versatility, } performance etc. you might get a long way towards a solution with a } simpler camera like the Nikon Coolpix 4500. It has remarkable macro } capability for the price, focussing to about 1 inch from front of } lens, and there is a so-called ringflash accessory for it too, } actually a ring of high-intensity white LEDs. I use one of these } myself, and know many other scientists who swear by them for } field-recording of specimens ranging from fluorescent gels to plants } and fungi, even aurora borealis displays are within its capabilities. }
I endorse the comments about the 4500. I have one, and it's great.
Unfortunately, the Coolpix 4500 seems to have been discontinued, at least in this country.
Does anyone know why, or whether Nikon is planning to launch a successor?
Anyone from Nikon care to reply?
cheers
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Sun Oct 10 16:18:07 2004
Curious! It is still listed on the Nikon Europe site. http://www.europe-nikon.com/details.aspx?countryid=20&languageid=22&prodId=117&catId=76 I presume any replacement would be introduced into US first. With 8megapixels perhaps. Chris
Dr. Chris Jeffree University of Edinburgh
----- Original Message ----- } From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} To: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} ; {microscopy-at-msa.microscopy.com} Sent: Sunday, October 10, 2004 9:45 PM
Hello everyone,
I am currently involved in a research project that is using many different imaging modalities to look at objects of a variety of sizes. I am interested in using texture parameters to try to classify this material and was wondering if there was any existing computer programs or relevant papers written that could help my with my project.
Please feel free to email me if you think you can help. Thank you all for your time.
Thomas Sadowski Southern Connecticut State University
_________________________________________________________________ Don’t just search. Find. Check out the new MSN Search! http://search.msn.click-url.com/go/onm00200636ave/direct/01/
From MicroscopyL-request-at-ns.microscopy.com Sun Oct 10 18:00:56 2004
In response to my posting about an hour ago regarding the unavailability of the Coolpix 4500, I have so far received Out of Office Autoreplies from Ian Kitajama, Robert Hull, Jonathon Dunlap, Tim Maitland, Pei Zou, and John Czernawski, plus an interesting one apparently generated by antispam software at the office of videbula-at-uol.com.br, which invited me to respond so that it would not classify my posting as spam!
Guys, can't you understand that by using auto reply you are threatening the continued existence of this forum?
If everyone who makes a posting knows that their inboxes are going to fill up with autoreplies, people are going to think twice before posting.
Please have the courtesy to either unsubscribe for the duration of your absences, or to susbscribe to the list using an alternative email address.
thanks
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Sun Oct 10 19:06:32 2004
My local Nikon agent says that production of this great model has now ceased, so when stocks run out, that's it.
So if you're contemplating buying one and if they're still available in your country, buy it now.
cheers
rtch
} From: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} To: "Ritchie Sims" {r.sims-at-auckland.ac.nz} Copies to: {microscopy-at-msa.microscopy.com}
A solution might be the Nikon CoolPix 8700. It has the following technical data:
NIKON COOLPIX 8700 DIGITAL CAMERA with 8.0 effective megapixels and an 8x Zoom-Nikkor ED lens Large vari-angle 270° highly transmissible advanced TFT-LCD monitor for greater visibility - even in daylight
We are offering high quality adapters for digital cameras, also for the Nikon 8700, so that you can connect it to a microscope (eyepiece or phototube). If you would like to know more about the adapters or if you wish to receive a list of adapters we offer, please contact me.
RS} ------------------------------------------------------------------------------ RS} The Microscopy ListServer -- Sponsor: The Microscopy Society of America RS} To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver RS} On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html RS} -------------------------------------------------------------------------------
} } } } Depending on how critical you are of issues like camera versatility, } } performance etc. you might get a long way towards a solution with a } } simpler camera like the Nikon Coolpix 4500. It has remarkable macro } } capability for the price, focussing to about 1 inch from front of } } lens, and there is a so-called ringflash accessory for it too, } } actually a ring of high-intensity white LEDs. I use one of these } } myself, and know many other scientists who swear by them for } } field-recording of specimens ranging from fluorescent gels to plants } } and fungi, even aurora borealis displays are within its capabilities. } }
RS} I endorse the comments about the 4500. I have one, and it's great.
RS} Unfortunately, the Coolpix 4500 seems to have been discontinued, at least in this RS} country.
RS} Does anyone know why, or whether Nikon is planning to launch a successor?
RS} Anyone from Nikon care to reply?
RS} cheers
RS} rtch
RS} -- RS} Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 RS} Microanalyst Fax : 64 9 3737435 RS} Department of Geology email : r.sims-at-auckland.ac.nz RS} The University of Auckland RS} Private Bag 92019 RS} Auckland RS} New Zealand
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 11 07:45:26 2004
A post-doctoral position is available January 2005. This position requires extensive hands-on experience in transmission/analytical electron microscopy, scanning electron microscopy, x-ray diffraction, and other analytical characterization techniques. Additional experience in materials synthesis and/or devices is highly desirable. Please contact: Prof. Nitin Padture {nitin.padture-at-uconn.edu} .
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Nitin P. Padture, Ph.D. Professor Department of Materials Science and Engineering Institute of Materials Science 97 N. Eagleville Road University of Connecticut, Storrs, CT 06269-3136, USA Phone: (860)486-4206; FAX: (860)486-4745 Email: nitin.padture-at-uconn.edu Website: http://www.ims.uconn.edu/metal/faculty/padture.htm ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 11 13:35:43 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dmwilliams-at-dow.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, October 11, 2004 at 13:25:07 ---------------------------------------------------------------------------
Email: dmwilliams-at-dow.com Name: David M. Williams
Question: Does anyone have a fixation procedure for the whole body perfusion of rabbits? (adult New Zealand White). (We only have experience with rats).
I would like to hear from anyone who has bought a diamond knife in the past couple of years. I would like to know your experiences with the knife's quality. I would like feedback about the following brands; Microstar, Diatome, and Delaware Diamond. Please reply to me privately.
Tom Bargar Univ. Nebraska Medical Center Core Electron Microscopy Research Facility tbargar-at-unmc.edu 402-559-7347
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 12 08:33:49 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jwyffel-at-mindspring.com) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, October 12, 2004 at 08:31:03 ---------------------------------------------------------------------------
Question: I am searching for a stain/technique to use on shark eggs. There is a carbohydrate jelly found within the egg case and surrounding the ovum much like albumen in chicken eggs. It varies in density or viscosity with location. I would like to image the differences using a carbohydrate stain that would hopefully stain the regions differentially. I will bisect the egg case before staining. Any ideas or suggestions are welcome. Thanks Jennifer
Does anyone out there know what kinds of microscopy are used in evaluating animal feed for contaminants or pathogens? It seems to me that optical microscopy would be as valuable a tool as TEM/SEM but I am unsure what role EDX plays in analysis of animal feed.
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 12 14:44:43 2004
Does anyone have contact information for Consolidated Vacuum Corp., or know that they no longer exist? I cannot find contact information for them, although searching the web did provide a spring 2000 dividend statement and lots of used CVC equipment. I have an ancient CVC diffusion pump for which I need information. The odd thing about this pump is that the heater is internal and sits within the bottom of the tree, in direct contact with the oil. No normal external pancake heater. If anyone has information about such a diff pump, I would appreciate hearing about it. Thanks. -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 12 21:09:43 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mburnham-at-kentdenver.org) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, October 12, 2004 at 14:28:01 ---------------------------------------------------------------------------
Email: mburnham-at-kentdenver.org Name: Michael Burnham
Organization: Kent Denver School
Education: 9-12th Grade High School
Location: Denver, Colorado
Question: We have an old but fully functional Zeiss EM9 transmission electron microscope, which has developed a "flicker" or pulse in the beam. This intermittent pulse momentarily increases the intensity of the beam which ruptures the specimen. I've tried various remedies---replace the filament, checking contacts and tubes, etc. but to no avail. I am about to replace the gun with an old-stock spare; however, I'd truly appreciate suggestions (including if anyone knows of a technician in the Denver area who still might work on old teaching TEM's).
Thank you, Michael Burnahm Science Chair Kent Denver School Englewood, CO 80113 303-770-7660 ex. 203
In the past few years I have used this mailing lists to discuss numerous microscopy issues and I have learned a lot from the people who are a member of this community.
This mailing list is mostly used by people who work in microscopy and who have a background in using a microscope in a "traditional" way. Most people know about Koehler illumination, the meaning of N.A. for resolution when doing microscopy with the naked eye. For digital microscopy, most people have a basic understanding of Nyquist sampling and the spatial requirements for image quantification. Young scientist always get help from the community on these issues.
Traditional people using a microscope in biology had a background in biology, medicine, etc. and some basic understanding of morphology and shape. Microscopy and morphology were a bit out of focus due to the emphasis on genomics and proteomics, but HCS brought it back. However with the advent of High Content Screening (HCS), biochemists started using devices based on optical microscopes without any basic training in (digital) optical microscopy or morphology. Studying a cell or tissue requires some basic knowledge of morphology and the interaction of light with matter and the optics of a microscope.
Modern software design sort of "uncouples" the HCS world from the optical reality of the optical instrument, but basicaly these machines are optical microscopes and adhere to the same optical principles as a "traditional" microscope. In my opinion using such an instrument without any basic understanding op digital optical microscopy makes no sense.
My question is, how do microscopists who are trained in "classical" microscopy explain the basics of microscopy to biochemists ? Are there textbooks or educational books written on biological optical microscopy with the non-morphologist in mind ?
Regards,
Peter
P.S I do not want to offend anyone, but it is something I experienced personally and I am at a loss how to deal with it.
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 13 07:32:42 2004
I have a Zeiss EM900 and have/had a similar problem. To eliminate the flicker I changed the objective aperture to the next larger one (50um ?). Our Zeiss repair person thinks the smaller aperture was being contaminated too quickly and causing a charge build up. The high voltage was checked and no variations were noted. Since the larger aperture, the flicker has not reappeared.
Best of luck,
Ed
Edward Calomeni Director EM Lab Ohio State University - Pathology M018 Starling Loving Hall 320 W. 10th Ave. Columbus, OH 43210-1240 614-293-5580 (office) 614-293-8806 (lab) calomeni-1-at-medctr.osu.edu
} } } by way of Ask-A-Microscopist {mburnham-at-kentdenver.org} 10/12/2004 10:13:13 PM } } }
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mburnham-at-kentdenver.org) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, October 12, 2004 at 14:28:01 ---------------------------------------------------------------------------
Email: mburnham-at-kentdenver.org Name: Michael Burnham
Organization: Kent Denver School
Education: 9-12th Grade High School
Location: Denver, Colorado
Question: We have an old but fully functional Zeiss EM9 transmission electron microscope, which has developed a "flicker" or pulse in the beam. This intermittent pulse momentarily increases the intensity of the beam which ruptures the specimen. I've tried various remedies---replace the filament, checking contacts and tubes, etc. but to no avail. I am about to replace the gun with an old-stock spare; however, I'd truly appreciate suggestions (including if anyone knows of a technician in the Denver area who still might work on old teaching TEM's).
Thank you, Michael Burnahm Science Chair Kent Denver School Englewood, CO 80113 303-770-7660 ex. 203
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rangari0-at-yahoo.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, October 12, 2004 at 22:51:23 ---------------------------------------------------------------------------
You should use diamond abrasive paper to ground the sample to about 40microns, then ion-mill the sample to electron transparent!
Changhui
by way of MicroscopyListserver wrote:
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From MicroscopyL-request-at-ns.microscopy.com Wed Oct 13 16:06:43 2004
You don't mention your substrate. I assume that this is a diamond film on a substrate such as silicon.
If your film is on a Si substrate, then I highly recommend the small angle cleavage technique or Microcleave technique. Check the South Bay Technology website for their microcleave kit and associated PDF files by John McCaffrey and myself.
If you must do ion milling, then you should be aware that carbon films do not ion mill very well with Ar beams. However, if you have a low angle mill, then things are alleviated quite a lot. One of the things that people have done to overcome the low milling rate of carbon is to add a little O2 into the Ar. Try 20%.) This is essentially a reactive etch for you. I have a paper in the MRS number four series on TEM sample prep that shows the relative differences of ion milling rates with Ar, Ne, Ar+O2, and Ne+O2 gases in a Gatan Duomill at 10 degrees. We used AFM to measure the relative heights of a DLC film on a TiC film on a silicon substrate. The idea was to try to match the mass of both the C and Si with the gas as close as possible. The best results were obtained using Ne+O2. Ne did a good job also.
-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)
-----Original Message----- } From: rangari0-at-yahoo.com [mailto:rangari0-at-yahoo.com] Sent: Wednesday, October 13, 2004 9:44 AM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rangari0-at-yahoo.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, October 12, 2004 at 22:51:23 ---------------------------------------------------------------------------
Is there anyone that can point me in the direction of some papers that show the modeling of EELS spectral lines from the density of states calculations for crystalline oxides. I am particularly interested in TiO2. If you know of any programs that are available, I would also be interested in that.
-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)
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 14 09:21:47 2004
Well, here I am pulling my hair out again, trying to get things to work as I would like.. I am currently trying to get TEM images in 16-bit Digital Micrograph 3 format into 8-bit tiff format - which I need to do since the images are held centrally on a server, and need to be read by lots of internal customers using their own PCs. I have at my disposal PhotoShop 4.0 and 5.0 with the Reindeer Games image processing toolkit, Thumbs Plus 4.5, as well as ImageJ and Irfan view.
The problem - only a small amount of the 16-bit dynamic range is present in the image, and the levels used vary from image to image. 'Auto levels' in PhotoShop doesn't work as well as I thought it would, since it goes too far and saturates the bright and dark pixels. I can adjust things as I would like manually using PhotoShop(minimum used level = 0, maximum used value = 65535, then convert to 8-bit), but this is pretty tedious to do for every image. Nothing else seems to be able to do what I want. I can't be the first person to have this problem - so does anyone know of a solution (preferably a free one, of course)?
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From MicroscopyL-request-at-ns.microscopy.com Thu Oct 14 10:31:09 2004
In more recent versions of Photoshop, and in the Reindeer Graphics Optipix plugins, you can set the amount of "tail clipping" that occurs - the fraction of the pixels that are set to black and white by adjusting the limits on your image. That feature was not present in Photoshop 4 or 5. Probably the best (fast, relatively cheap) approach would be to upgrade to the latest Photoshop, and then create a simple action that opens an image, auto-adjusts the levels with a small tail clip (e.g., 0.01% at each end), converts to 8 bits, and saves the image into a new folder. That can be run as a batch while you do something else, and will produce a new set of images for your server without replacing the originals,just in case someone needs the full 16 bit dynamic range or the actual original grey scale values later on.
John Russ ====== In a message dated 10/14/04 11:22:52 AM, richard.beanland-at-bookham.com writes:
} ... {snip} I am currently trying to get TEM images in 16-bit Digital } } Micrograph 3 format into 8-bit tiff format - which I need to do since the } } images are held centrally on a server, and need to be read by lots of } } internal customers using their own PCs. I have at my disposal PhotoShop } 4.0 } } and 5.0 with the Reindeer Games image processing toolkit, Thumbs Plus 4.5, } } as well as ImageJ and Irfan view. }
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 14 10:52:26 2004
I think I might have the record for a problem solved by the ListServer - 7 minutes from posting to solution! Many thanks indeed John, and unabashed adoration for Nestor for running this great resource..
-----Original Message----- } From: DrJohnRuss-at-aol.com [mailto:DrJohnRuss-at-aol.com] Sent: 14 October 2004 16:35 To: richard.beanland-at-bookham.com; microscopy-at-msa.microscopy.com
I would like to know if anyone has experience in getting a Digital Single Lens Reflex camera (DSLR) to provide a "live" preview for display on a Computer.
We purchased a Nikon D-70 and Nikon Image Capture 4.1 software which allows remote control of the camera through a PC. After initial tests it seems that it is impossible to acquire even a low resolution "live" preview with this camera.
Capturing an image through the computer interface of Nikon Capture 4.0 is simple but the lack of a preview (even low-res) is quite inconvenient.
In theory it should be possible for the camera to be controlled such that the mirror is held in the open position and the CCD continuously dumps the data to the PC. This method doesn't seem like something that is supported by standard OEM provided solutions or is as easy as we'd first hoped.
Our intermediate solution is to place a crosshair occular on the scope which will be used to make the image and camera parfocal before simply taking a picture. While it works we'd really like to get a live preview and make it ridiculously simple to use (which was the reason for picking this camera initially.)
If anyone has any suggestions or has experienced this problem I'd greatly appreciate your help.
Thanks, Dustin Grzesik
Research Technician Analytical Imaging Facility Albert Einstein College of Medicine
-- "Strong, light, cheap. pick two." Keith Bontrager
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 14 14:22:02 2004
Richard Auto-level function in the Photoshop works fine to me for most DM images. I also use macros (action) function to convert many files altogether as Jon Russ suggested. Auto-level function does not work well if you have very bright or dark area on your picture, then you have to adjust levels manually (and I don't think you could use automatization there). I also find that DM did good job choosing B&W balance when convert images into 8-bit using export-} tiff option. So, I usually create two sets of images (16 and 8-bit) at the level of DM, not Photoshop. The problem with DM is that I have difficulties to automate the process. DM has quite extended macros language and it supposed to be quite easy to write macros for automatic conversion 16 into 8-bit and save into another directory. But, perhaps, I am too lazy to do it well, so it does not work in my hands. If somebody already crossed that border and has functioning macros and willing to share, I would be grateful to have it. Have a good day, Sergey.
At 03:22 PM 10/14/2004 +0100, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Thu Oct 14 16:05:01 2004
Although it may not be too helpful for PC users, the incredibly low-cost "Graphics Converter" program which I run on my mac does a good job reading the DM files for me. It also has a batch function for converting them all at once.
Stuart
On Oct 14, 2004, at 2:27 PM, Sergey Ryazantsev wrote:
} } } ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } Richard } Auto-level function in the Photoshop works fine to me for most DM } images. I also use macros (action) function to convert many files } altogether as Jon Russ suggested. Auto-level function does not work } well if you have very bright or dark area on your picture, then you } have to adjust levels manually (and I don't think you could use } automatization there). I also find that DM did good job choosing B&W } balance when convert images into 8-bit using export-} tiff option. So, } I usually create two sets of images (16 and 8-bit) at the level of DM, } not Photoshop. The problem with DM is that I have difficulties to } automate the process. DM has quite extended macros language and it } supposed to be quite easy to write macros for automatic conversion 16 } into 8-bit and save into another directory. But, perhaps, I am too } lazy to do it well, so it does not work in my hands. If somebody } already crossed that border and has functioning macros and willing to } share, I would be grateful to have it. Have a good day, Sergey. } } At 03:22 PM 10/14/2004 +0100, you wrote: } } } } ---------------------------------------------------------------------- } } -------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------- } } --------- } } } } Well, here I am pulling my hair out again, trying to get things to } } work as I } } would like.. I am currently trying to get TEM images in 16-bit } } Digital } } Micrograph 3 format into 8-bit tiff format - which I need to do since } } the } } images are held centrally on a server, and need to be read by lots of } } internal customers using their own PCs. I have at my disposal } } PhotoShop 4.0 } } and 5.0 with the Reindeer Games image processing toolkit, Thumbs Plus } } 4.5, } } as well as ImageJ and Irfan view. } } } } The problem - only a small amount of the 16-bit dynamic range is } } present } } in the image, and the levels used vary from image to image. 'Auto } } levels' } } in PhotoShop doesn't work as well as I thought it would, since it } } goes too } } far and saturates the bright and dark pixels. I can adjust things as } } I } } would like manually using PhotoShop(minimum used level = 0, maximum } } used } } value = 65535, then convert to 8-bit), but this is pretty tedious to } } do for } } every image. Nothing else seems to be able to do what I want. I } } can't be } } the first person to have this problem - so does anyone know of a } } solution } } (preferably a free one, of course)? } } } } Many thanks in advance } } } } Richard } } } } _______________________________ } } Richard Beanland } } Analytical Services } } Bookham Inc } } Caswell, } } Towcester, } } Northamptonshire NN12 8EQ } } UK } } Tel: +44 (0) 1327 356362 } } Fax: +44 (0) 1327 356775 } } 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. } } ====================================================================== } } = } }
Prof. Stuart McKernan IT Characterization Facility, University of Minnesota E-mail : stuartm-at-umn.edu 12 Shepherd Labs, Office: (612) 624-6009 100 Union Street S. E., Minneapolis, MN 55455 Lab: (612) 626-7594
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 14 16:34:48 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (monica.iliescu-at-polymtl.ca) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, October 14, 2004 at 16:30:39 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] QX-102 capsules for ESEM
Question: Dear colleagues,
I would like to learn if someone of you use QX-102 capsules for ESEM imaging of wet samples (namely, biological) at atmospheric pressure. QuantomiX Inc manufactured like capsules in disposable package. My question is if these capsules can be used again after one experiment, i.e. if they can be used subsequently for many samples when sterile conditions are not imposed.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (breath999-at-yahoo.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, October 13, 2004 at 14:47:24 ---------------------------------------------------------------------------
Email: breath999-at-yahoo.com Name: Peter Bradley
Organization: UCLA
Title-Subject: [Microscopy] [Filtered] MListserver: Formaldehyde/PBS - old works better?
Question: I'm hoping someone here can help me out with storage of formaldehyde in PBS. I found out a person here was using fixative (3.7% formaldehyde in PBS) stored in the fridge for about 2-3 weeks. I told him to toss it out and only use fresh. It turns out that staining (IFA) with one antibody is beautiful in 2 week old but destroyed in fresh fix. We have recently tried various titrations of % formald, time, etc. but none give the nice staining seen in 2 week old formald (1 week is not long enough). I have seen a bit about polymerization, etc, but I am concerned that an artifact could be created by the old fix.
Any help would be greatly appreciated. Thanks in advance, Peter
} From: Day, Jeff Sent: Wednesday, October 13, 2004 10:29 AM To: 'Microscopy-at-MSA.Microscopy.Com'
Monica, They are single use items at about $40 a capsule. You don't need ESEM to use them, just a good backscatter detector. Gianni Torraca Analytical Sciences AMGEN
-----Original Message----- } From: monica.iliescu-at-polymtl.ca [mailto:monica.iliescu-at-polymtl.ca] Sent: Thursday, October 14, 2004 2:39 PM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (monica.iliescu-at-polymtl.ca) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, October 14, 2004 at 16:30:39 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] QX-102 capsules for ESEM
Question: Dear colleagues,
I would like to learn if someone of you use QX-102 capsules for ESEM imaging of wet samples (namely, biological) at atmospheric pressure. QuantomiX Inc manufactured like capsules in disposable package. My question is if these capsules can be used again after one experiment, i.e. if they can be used subsequently for many samples when sterile conditions are not imposed.
Hello Marie-Claude, I have some chemists doing spectroscopy using an inverted microscope base in our facility. Ours is a 12V halogen lamp, but thier tests indicate a 4 hour warm up period for maximum stability. Having the lamp power supply rheostat adjusted to provide nearly highest output (highest color temperture) aids in warm up. Also, a new (high-quality) bulb might help in your case. Regards, Karl
Marie-Claude Bélanger wrote:
} } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } Dear All, } } I've recently started taking birefringence photographs of Congo Red } stained tissue. I've noticed that over time, the very same field } doesn't appear the same, and that the labeled surface varies up to 25%. } } Everything being very stable (all screws firmly screwed, polarizer } glued to its base), I'm wondering if anyone has performed a test to } check the warm-up period of a halogen 6 volt lamp. The company who } sells the lamp says 5 to 10 minutes, but could it be longer? } } Thank you! } } Marie-Claude Belanger } } _________________________________________________________________ } Des mécanismes de contrôle parental puissants permettent à votre } enfant de découvrir tout ce qu’Internet a à offrir. } http://join.msn.com/?pgmarket=fr-ca&page=features/parental&ST=1&xAPID=1983&DI=2043 } Commencez dès maintenant à profiter de tous les avantages de MSN } Premium et obtenez les deux premiers mois GRATUITS*. }
-- Karl Garsha Light Microscopy Specialist Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana-Champaign 405 North Mathews Avenue Urbana, IL 61801 Office: B650J Phone: 217.244.6292 Fax: 217.244.6219 Mobile: 217.390.1874 www.itg.uiuc.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 15 12:03:59 2004
I am one of those stubborn Mac users and I need an advise in dealing with Gatan DigitalMicrograph images.
We use DigitalMicrograph v.3.7 on a Windows PC to acquire TEM images. I have DigitalMicrograph v.3.4 on a Mac and I want to use my Mac to work with the image files.
The problem is that the Mac DM3.4 does not recognize the .dm3 file format from the PC. Usually I save two versions of each file one in .dm3 format and one in TIF, which I can use on the Mac. The problem is that this is a time consuming procedure when dealing with a large number of files.
I have other software such as ImageJ and Graphic Converter on my Mac and I can open the .dm3 files with them. The problem is that the annotation is lost during the transfer.
The question: Is there a way to convert multiple files as batch process from dm3 into TIF format without opening and saving each file individually, where the original annotation in the files and the file names will remain unchanged?
Thanks,
Krassimir.
P.S. Switch to PC is not regarded as a valid answer.
--------------------------------------------------------- Krassimir N. Bozhilov, PhD EM Scientist and Manager Central Facility for Advanced Microscopy and Microanalysis University of California Riverside, CA 92521
Tel 951 827 2998 Fax 951 827 2489 --------------------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 15 13:32:39 2004
I don't think formalin in PBS would 'go bad' or lose much activity in 2 weeks in the 'frig. Even formaldehyde made fresh from paraformaldehyde -which I was always taught must be made fresh- lasts for several weeks in the cold. That was published in the last few years but I can't find the reference, if anyone has it please send it to me. There are several variables we need to know before we can make an informed decision. How long was the tissue fixed (formalin works slowly)? How old was the formaldehyde used to make the fix-PBS mixture? Was the stock bottle already weakened by polymerization of formalin? Fixed in the cold or room temp? What kind of tissue? Hoiw about the rest of the processing? What antibody and detection system? How many trials did you do with each fix? It sounds like your sample size for the 'old, cold' fix is one, hardly sufficient to draw a valid conclusion.
Geoff
by way of MicroscopyListserver wrote:
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-- -- ********************************************** 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 **********************************************
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 15 13:53:51 2004
On Oct 15, 2004, at 10:08 AM, K.N. Bozhilov wrote:
} I am one of those stubborn Mac users and I need an advise in dealing } with } Gatan DigitalMicrograph images. } } We use DigitalMicrograph v.3.7 on a Windows PC to acquire TEM images. } I have DigitalMicrograph v.3.4 on a Mac and I want to use my Mac to } work } with the image files. } } The problem is that the Mac DM3.4 does not recognize the .dm3 file } format } from the PC. Usually I save two versions of each file one in .dm3 } format and } one in TIF, which I can use on the Mac. The problem is that this is a } time } consuming procedure when dealing with a large number of files. } } I have other software such as ImageJ and Graphic Converter on my Mac } and I } can open the .dm3 files with them. The problem is that the annotation } is } lost during the transfer. } } The question: Is there a way to convert multiple files as batch } process from } dm3 into TIF format without opening and saving each file individually, } where the original annotation in the files and the file names will } remain } unchanged? } Dear Krassimir, Could you please post the responses you get so us other stubborn Mac users can convert our files? If this is done using a GATAN script in DM, could it also be used to convert the data type to unsigned, 2-byte? TIA. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 15 15:37:09 2004
We have found that many of the standard power supplies provided by the microscope manufacturers are noticeably unstable as measured with tube or CCD camera time lapse imaging of cells by phase contrast, at least with the power supply from the wall of 102 to 118 VAC here in NYC.
On the microscope stations where we have needed to rectify this, we have purchased special stable power sources (AC to 12V DC conversions) instead of using the ones supplied in the bases of the microscopes.
-Michael
On Fri, 15 Oct 2004, Karl Garsha wrote:
} Hello Marie-Claude, } I have some chemists doing spectroscopy using an inverted microscope } base in our facility. Ours is a 12V halogen lamp, but thier tests } indicate a 4 hour warm up period for maximum stability. Having the lamp } power supply rheostat adjusted to provide nearly highest output (highest } color temperture) aids in warm up. Also, a new (high-quality) bulb might } help in your case. } Regards, } Karl } } Marie-Claude Bélanger wrote: } } } Dear All, } } } } I've recently started taking birefringence photographs of Congo Red } } stained tissue. I've noticed that over time, the very same field } } doesn't appear the same, and that the labeled surface varies up to 25%. } } } } Everything being very stable (all screws firmly screwed, polarizer } } glued to its base), I'm wondering if anyone has performed a test to } } check the warm-up period of a halogen 6 volt lamp. The company who } } sells the lamp says 5 to 10 minutes, but could it be longer? } } } } Thank you! } } } } Marie-Claude Belanger } } } } _________________________________________________________________ } } Des mécanismes de contrôle parental puissants permettent à votre } } enfant de découvrir tout ce qu’Internet a à offrir. } } http://join.msn.com/?pgmarket=fr-ca&page=features/parental&ST=1&xAPID=1983&DI=2043 } } Commencez dès maintenant à profiter de tous les avantages de MSN } } Premium et obtenez les deux premiers mois GRATUITS*. } } } } -- } Karl Garsha } Light Microscopy Specialist } Imaging Technology Group } Beckman Institute for Advanced Science and Technology } University of Illinois at Urbana-Champaign } 405 North Mathews Avenue } Urbana, IL 61801 } Office: B650J } Phone: 217.244.6292 } Fax: 217.244.6219 } Mobile: 217.390.1874 } www.itg.uiuc.edu } } } }
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 15 18:31:32 2004
We just got a spiffy new inverted light microscope. It is in a multiple user lab and most will want to use a 63X oil objective.
Most of my experience is with standard upright scopes where adding the oil is pretty up front and easy, slide stays on the stage, swing objective out, add oil, swing objective back in. This inverted thing is a little more complicated.
Some users get the objective in position, remove their slide, add oil, then replace the slide. Works OK, but seems like slide might not land back in exactly the same spot and a once in a lifetime location might be lost.
So, any advice or clever techniques. I have thought of a long probe to reach under the slide, but could be a mess. Have read some advice about rotating objective to the side, adding oil, then rotating back quickly before oil slips off.
Not a big deal, but seems like there should be a better way.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 15 18:35:50 2004
Dear Krassimir and scripters, I have the same problem with the DM for Mac. I have found that converting to DM2 format will allow me to open my DM3-PC files on the Mac. Also, for full 16-bit data, I have found the gfx format to be quite portable. Gatan is constantly changing how it saves 16-bit TIF files that you never know what you are going to get. The Gatan script below will convert a directory full of DM3 files to DM2 format. It does so by using the "ImageDisplayExportToFile" command. By changing a few variables (see script) it will save in any format Gatan supports.
I have found that these script files do not cut and paste well from email. You may need to clean up the line breaks a bit to get it to work correctly. If necessary, I can send you the script as an attachment.
Hope this helps, Ray
/* BatchDirFileConvert.s
The basic idea is to scan through a directory for DM files and convert them to a new format The script uses the "void ImageDisplayExportToFile( ImageDisplay imgDisp, String format, String file_name ) " script command. Here you give the export format in the string "format". Gatan changes how this works from version to version so use at your own risk.
Copyright 10/15/2004 Ray D. Twesten - University of Illinois
*/
//****************************** //Modify these strings to change format. // See SaveAs or Export menu item for correct strings //****************************** string format = "Gatan 2.5 Format" // Good for sending to Mac version of DM string extension = ".dm2" string saveDir = "DM2"
// // string format = "Fixed Format" // Good all around 16-bit format // string extension = ".gfx" // string saveDir = "GFX"
// // Get image path and create tag group to hold file names: // string fileName = "" if(!OkCancelDialog("Choose Any file in Dir to Open")) exit(0) if(!OpenDialog(fileName)) exit(0) string dirName = PathExtractDirectory( fileName, 0 )
Taggroup dir = GetFilesInDirectory( dirName, 1 ) number numfiles = TagGroupCountTags( dir )
// End BatchDirFileConv.s //**********************************************************8
Ray D. Twesten, PhD. Center for Microanalysis of Materials Seitz Materials Research Laboratory 104 S. Goodwin Ave., Urbana, IL 61801 +1 217 244-6177 (Fax -2278)
-----Original Message----- } From: K.N. Bozhilov [mailto:bozhilov-at-ucr.edu] Sent: Friday, October 15, 2004 12:08 PM To: microscopy-at-msa.microscopy.com
Hi,
I am one of those stubborn Mac users and I need an advise in dealing with Gatan DigitalMicrograph images.
We use DigitalMicrograph v.3.7 on a Windows PC to acquire TEM images. I have DigitalMicrograph v.3.4 on a Mac and I want to use my Mac to work with the image files.
The problem is that the Mac DM3.4 does not recognize the .dm3 file format from the PC. Usually I save two versions of each file one in .dm3 format and one in TIF, which I can use on the Mac. The problem is that this is a time consuming procedure when dealing with a large number of files.
I have other software such as ImageJ and Graphic Converter on my Mac and I can open the .dm3 files with them. The problem is that the annotation is lost during the transfer.
The question: Is there a way to convert multiple files as batch process from dm3 into TIF format without opening and saving each file individually, where the original annotation in the files and the file names will remain unchanged?
Thanks,
Krassimir.
P.S. Switch to PC is not regarded as a valid answer.
--------------------------------------------------------- Krassimir N. Bozhilov, PhD EM Scientist and Manager Central Facility for Advanced Microscopy and Microanalysis University of California Riverside, CA 92521
Tel 951 827 2998 Fax 951 827 2489 --------------------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 15 19:51:17 2004
I am using an inverted fluorescent microscope and I do not seem to have this kind of problems... I usually put the oil before I place the slide.
I however have enough space around the objective to swing it out just a little bit, add the oil with no risk of slipping off and than swing it back in. To make my life easier, I also have a sort of opening on the stage that is made to add the oil.
I can send pictures of my stage if you want.
Hope that helps,
Anne-Laure
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Le Ny Anne-Laure Grad Student Chemical engineering University of Southern California PCE 310 Tel: 213-740-1320 Cell: 626-840-5456 ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
On Fri, 15 Oct 2004, Jon Krupp wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Greetings: } } We just got a spiffy new inverted light microscope. It is in a multiple } user lab and most will want to use a 63X oil objective. } } Most of my experience is with standard upright scopes where adding the oil } is pretty up front and easy, slide stays on the stage, swing objective out, } add oil, swing objective back in. This inverted thing is a little more } complicated. } } Some users get the objective in position, remove their slide, add oil, then } replace the slide. Works OK, but seems like slide might not land back in } exactly the same spot and a once in a lifetime location might be lost. } } So, any advice or clever techniques. I have thought of a long probe to } reach under the slide, but could be a mess. Have read some advice about } rotating objective to the side, adding oil, then rotating back quickly } before oil slips off. } } Not a big deal, but seems like there should be a better way. } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } } }
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 16 02:32:45 2004
: : We have found that many of the standard power supplies provided by the : microscope manufacturers are noticeably unstable as measured with tube or : CCD camera time lapse imaging of cells by phase contrast, at least with : the power supply from the wall of 102 to 118 VAC here in NYC. : : On the microscope stations where we have needed to rectify this, we have : purchased special stable power sources (AC to 12V DC conversions) instead : of using the ones supplied in the bases of the microscopes. : : -Michael : : On Fri, 15 Oct 2004, Karl Garsha wrote: : : } Hello Marie-Claude, : } I have some chemists doing spectroscopy using an inverted microscope : } base in our facility. Ours is a 12V halogen lamp, but thier tests : } indicate a 4 hour warm up period for maximum stability. Having the lamp : } power supply rheostat adjusted to provide nearly highest output (highest : } color temperture) aids in warm up. Also, a new (high-quality) bulb might : } help in your case. : } Regards, : } Karl : } For low voltage bulbs I have solved this problem in the past with a lead acid auto battery on a trickle charger with a resistor to reduce the voltage to what ever I need on the lamp. Then you only have to wait until the resistor heats up until the light is stable. It also gets rid of the very slight 120 Hz flicker on the bulb if you are using AC.
The flicker is not normally a problem as no camera will pick it up and your eyes will never see it but it can be in some imaging if the capture is fast enough.
For 110 VAC I have generaly found the unconditioned power better than that going thourgh so called power conditioners. Put some surge protectors accorss the line and try it as is.
Gordon Gordon Couger
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others. Microscope Manual at www.science-info.org
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 16 12:51:57 2004
I've found the "move the objective" to be the best way to oil lens on an inverted 'scope. But, something else you might want to do: go to someplace like WalMart or the like and buy a bag of little hair scrunchies. They just fit the lenses, and even come color-coded. This will keep the oil from running down the lenses into threads, the turret, and so forth.
Phil
} Greetings: } } We just got a spiffy new inverted light microscope. It is in a multiple } user lab and most will want to use a 63X oil objective. } } Most of my experience is with standard upright scopes where adding the oil } is pretty up front and easy, slide stays on the stage, swing objective out, } add oil, swing objective back in. This inverted thing is a little more } complicated. } } Some users get the objective in position, remove their slide, add oil, then } replace the slide. Works OK, but seems like slide might not land back in } exactly the same spot and a once in a lifetime location might be lost. } } So, any advice or clever techniques. I have thought of a long probe to } reach under the slide, but could be a mess. Have read some advice about } rotating objective to the side, adding oil, then rotating back quickly } before oil slips off. } } Not a big deal, but seems like there should be a better way. } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.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)
From MicroscopyL-request-at-ns.microscopy.com Sun Oct 17 11:42:41 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (diller-at-stefan-diller.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, October 17, 2004 at 07:48:38 ---------------------------------------------------------------------------
Email: diller-at-stefan-diller.com Name: Stefan Diller
Question: Is there anyone out there who can give me advice in using an EDAX 9100 system on a Philips EM 420? Especially I am looking for tips how to do linescans or element mapping... My manual is not so clear in this point.
The system has a digital ratemeter 9201 which - I understand - es necessary to aquire linescans and element-maps.
Does anybody know what the latest available software version for this system is and if there is any possibility to use "modern" printers for printing spectra and other data?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (innap-at-savion.huji.ac.il) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, October 17, 2004 at 11:23:10 ---------------------------------------------------------------------------
Question: Dear All, I would appreciate any help and recommendations in preparation of metallographic sample from the extruded polymer wire of 0.3-0.5 mm dia. The polymer wire is a composite material: polyethylene matrix strenghthened by ultra high molecular weight polyethylene fibers. The purpose of the study is to see the interface between the fibers and the matrix. Do you have any idea of how to polish cross sectional sample (I need them in two orthogonal directions) from this kind of material? Thank you in advance, Inna
The new version of Digital Micrograph 3.9 (GMS1.4) can do batch conversion to tif, jpg, and bmp formats.
There is also a plug-in for older versions of DM which can be used to convert to the above formats and also to dm2 format.
Problems still remaining:
Batch conversion to dm2 deletes all annotations. Scale bar is not the issue since the information about the pixel size is still preserved.
Conversion to non-Gatan format burns the annotations in the image data and annotations cannot be separated or moved if desired. Also such conversions result in certain loss of data in general.
Saving directly to dm2 is similar to converting to tif or jpeg, with the advantage that no data loss occurs, but magnification information is lost and all annotations become permanent part of the image data.
Conclusion: there is no perfect alternative to using Digital Micrograph on a Windows PC and working with dm3 files.
Thank you for all inputs.
Krassimir.
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 18 16:17:01 2004
Our lab has routinely processed samples according to the methods posted at http://www.micro.wisc.edu/smith in the past without artifacts, but recently we have encountered an unknown contaminant seen as an electron dense (opaque) amorphic precipitate. Please visit the web-site listed above for examples of the artifact and detailed processing methods. We would like your opinions as to the source of this contamination so we might avoid it in future experiments. Your comments are much appreciated.
Ben August U.W. Electron Microscopy Facility Medical School University of Wisconsin - Madison bkaugust-at-facstaff.wisc.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 18 16:19:22 2004
Could someone provide the digital equivalency (in terms of resolution) of 3.25 x 4 inch TEM film.
I have read that a 6 megapixel digital file has the resolution capability of a 35 mm grayscale negative. If this is true, then an 18 megapixel digital file would be equivalent to a TEM grayscale negative in terms of resolution capability.
I was thinking (based on darkroom enlargement capabilities of TEM films) that a 180 MP digital file would be more likely to exhibit the resolution one would see in a TEM negative.
Thanks for the information.
JB -- ############################################################## 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 Email: bozzola-at-siu.edu Web: http://www.siu.edu/~image/ ##############################################################
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 18 18:20:41 2004
Film with a fine grain size and lots of silver (e.g., TEM film) can resolve the equivalent of about 4000 points per inch ("pixels" if you like). Good film scanners can digitize film with that resolution. That means your 3.25x4 inch film would represent 13000 x 16000 pixels or about 2 x 10^8 pixels. But it would take twice that many bytes to hold the data because the dynamic range of film greatly exceeds 8 bits. Film with a lot of silver in it can easily produce 12 bits (1 part in 4000) and even a bit more. A MaxD of 4.2, which is possible with X-ray film, corresponds to about 14 bits. So you would have to store the data as two bytes per pixel. Pay no attention to the myth that a 6 megapixel camera delivers the performance of film - it isn't close in either dynamic range or spatial resolution.
John Russ ====== In a message dated 10/18/04 6:21:15 PM, bozzola-at-siu.edu writes:
} Could someone provide the digital equivalency (in terms of } resolution) of 3.25 x 4 inch TEM film. } } I have read that a 6 megapixel digital file has the resolution } capability of a 35 mm grayscale negative. If this is true, then an 18 } megapixel digital file would be equivalent to a TEM grayscale } negative in terms of resolution capability. } } I was thinking (based on darkroom enlargement capabilities of TEM } films) that a 180 MP digital file would be more likely to exhibit the } resolution one would see in a TEM negative. } } Thanks for the information. } } JB
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 18 19:17:48 2004
John - Assuming 7um silver grains that's about right - but, of course, it assumes that the point spread in the film is less than that and that each electron makes a single developable grain - neither of which is likely tom be achieved in reality. CCD and CMOS Cameras and imaging plates have ~15um pixels with higher QE than film. Optically coupled, as opposed to fiber optically coupled, CCDs offer very good MTF over a wide field of view. However, FO coupled CCDs have higher sensitivity - as low as single electrons. Imaging plates, on the other hand, provide extremely high dynamic range - far exceeding film or CCDs - while providing a wide field of view.
Bill Miller
At 05:26 PM 10/18/2004, John J. Bozzola wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 18 19:55:33 2004
On Oct 18, 2004, at 2:26 PM, John J. Bozzola wrote:
} Could someone provide the digital equivalency (in terms of resolution) } of 3.25 x 4 inch TEM film. } } I have read that a 6 megapixel digital file has the resolution } capability of a 35 mm grayscale negative. If this is true, then an 18 } megapixel digital file would be equivalent to a TEM grayscale negative } in terms of resolution capability. } } I was thinking (based on darkroom enlargement capabilities of TEM } films) that a 180 MP digital file would be more likely to exhibit the } resolution one would see in a TEM negative. } } Thanks for the information. } Dear John, Digital equivalency depends on, first, the grain size, and, second, the scanner pixel size. For most film, the grain size is small compared to the pixel size; however, for pixel size less than ~5 um, grain size may be important, and, as pixel size decreases below 5 um, grain size becomes increasingly important. A 3.25 x 4 inch film is roughly 16,000 x 20,000 5 um x 5 um pixels for a size of ~335 Mpixel, so, if you have a good scanner, 18 Mpixel does not contain nearly as much info as there is on a scan of a negative, and, furthermore, the negative itself has about 1.5 orders of magnitude more information than is retrieved by a good scanner (and even more, if you take into account that the ODs in the film may not be scanned quantitatively). High-resolution film, such as 4489, has even better resolution than these figures would indicate due to its extremely fine grain and relatively good OD range; SO163 is pretty much in line with these figures; LoDose X-ray film is somewhat poorer. The drawback to 4489 is that it takes more beam to get an image, and LoDose requires much less beam, so, since radiation damage is often limiting, it is sometimes best to try for less information with less damage--there is no free lunch. The equivalence of 6 Mpixel with 35 mm film may have been derived from assuming a certain print size and using the highest spatial frequency seen by the human eye. In other words, comparing a print from a 6 Mpixel file with one made from film, one would see no difference. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 19 00:46:53 2004
John If my arithmetic is correct, 3.25x4" EM negative contains approximately 3 Gb equivalent of digital information: standard film has 100-200 lines/mm resolution; technical films: 350-600 lines/mm. 600 lines/mm gives us 600*25=15000 dpi. 1 in^2=225*10^6 *3.25*4~3 Gb right? Not bad. Human eye produced 120 Mb gray (rods only) image and resolution is about 6000 dpi.
Color 35 mm film (24x36 mm) has resolution about 150 lines/mm, so: 24x36mm=864mm^2*150^2=19440000 ~19 Mpix
When you are trying to compare "analog" and digital photography one need to remember that we still don't have much instrumentation to handle that giant amount of information. I am not talking about computer recourses. See, for instance human eye may recognize only 200 shades of gray, so we actually could not see a difference between 8 and 16-bit gray images. Our monitors has normally 72 dpi resolution and this is what you are analyzing on the screen. Similarly, the published images has 72 dpi as well and barely reproduces 10-15 shades of gray... Sergey
At 02:26 PM 10/18/2004, 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 10833 Le Conte Ave, Room 33-080 Los Angeles, CA 90095
John Your estimate of 180mpixels is close to mine. Fine-grained b&w film can resolve ~80 lines per mm. Actually line pairs per mm, since a line is not a line unless seen against a background of different intensity. to resolve 80 lines you need 2 pixels per line: With 80 pixels per mm, 80 lines per mm merge into solid grey, so *a minimum* 160 pixels per mm required, assuming the spacing and position of the lines exactly corresponds with the pitch of the pixels. If not, again all you see is solid grey.
for 3.25x4" film = 13208 x 16256 pixels = 214.7 mega pixels for 35mm film = 5760x3840 = 22.1 mega pixels
Some high-performance films like Kodak Technical Pan can resolve better than 80 line pairs per mm
Note that for TEM film in particular you may need more than 8 bits to record the dynamic range of the film. Which is paradoxical, because at limiting resolution, film is a binary medium. A silver grain is there, or it isn't.
Chris
Dr. Chris Jeffree University of Edinburgh
----- Original Message ----- } From: "John J. Bozzola" {bozzola-at-siu.edu} To: {Microscopy-at-msa.microscopy.com} Sent: Monday, October 18, 2004 10:26 PM
The other issue to consider is the resolution of your microscope. It may be no worth scanning a negative to see individual silver grains, if your electron beam is printing at 100 grains wide.
Martin
At 05:58 AM 10/19/2004, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 19 08:18:45 2004
We are planning to buy a tissue processor for electron microscopy called "Leica EM TP" with its light microscopy processing attachment. Does anybody has experience with this stuff? Do you recommend using it? As you know, these are very expensive equipment. We want to learn advantages and disadvantages of working with it. For example is it cost effective comparing with manual method?
Thanks in advance...
Dr. Necat Yžlmaz Mersin Üniversitesi Tžp Fakültesi Histoloji ve Embriyoloji Anabilim Dalž 0 324 3413066
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 19 08:19:58 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (seversong-at-dpg.army.mil) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, October 18, 2004 at 17:26:52 ---------------------------------------------------------------------------
Email: seversong-at-dpg.army.mil Name: Grant Severson
Organization: U.S. Army Dugway Proving Ground
Education: Graduate College
Location: Dugway, UT
Question: I was wondering if you could recommend a good course in fluorescent microscopy techniques.
Benjamin, It looks like you had a Glut/OsO4/PO4 reaction. I would not osmicate in phosphate, in fact switch to 0.1 M cacodylate either before osmium or from primary fix. Also I would use(0.8% K4FeCN6) reduced (1%) osmium, you will get better myelin structure. I also advocate en-bloc 2% uranyl acetate before ethanol. Good Luck
Michael Delannoy Associate Director JHSM Microscope Facility Baltimore Md
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 19 11:51:12 2004
I think, all of these calculations come to approximately the same numbers for the total information content of a negative, something of the order of one to several hundred MBytes. I think, you are also right in that the calculations yield MBit instead of MByte due to the binary nature of the silver grains.
However, what we are talking of is the total information content of a negative. In the overwhelming number of cases, people don't need or don't even want this amount of information. Typical cases are Pathologists, who want to see the information on the entire negative, but don't care so much about the details embedded in the film at the grain level, or people who do high-resolution TEM. They are mostly interested in the highest resolution they can get, but not across the entire negative.
In my opinion (as a vendor of digital systems), the digital cameras have already surpassed film in terms of usability and other parameters. The total information content of film is higher, but to me that's like a newspaper: Some people want to read the comics, others the stock listings, a third person might prefer the political or foreign news. On a day to day basis, nobody really needs all the information that is in the newspaper.
And in the rare case that someone actually does need the full information content, you can always mosaic several TEM images into a large one.
Michael Bode
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: Chris Jeffree [mailto:c.jeffree-at-ed.ac.uk] Sent: Tuesday, October 19, 2004 02:59 To: John J. Bozzola Cc: microscopy-at-msa.microscopy.com
John Your estimate of 180mpixels is close to mine. Fine-grained b&w film can resolve ~80 lines per mm. Actually line pairs per mm, since a line is not a line unless seen against a background of different intensity. to resolve 80 lines you need 2 pixels per line: With 80 pixels per mm, 80 lines per mm merge into solid grey, so *a minimum* 160 pixels per mm required, assuming the spacing and position of the lines exactly corresponds with the pitch of the pixels. If not, again all you see is solid grey.
for 3.25x4" film = 13208 x 16256 pixels = 214.7 mega pixels for 35mm film = 5760x3840 = 22.1 mega pixels
Some high-performance films like Kodak Technical Pan can resolve better than 80 line pairs per mm
Note that for TEM film in particular you may need more than 8 bits to record the dynamic range of the film. Which is paradoxical, because at limiting resolution, film is a binary medium. A silver grain is there, or it isn't.
Chris
Dr. Chris Jeffree University of Edinburgh
----- Original Message ----- } From: "John J. Bozzola" {bozzola-at-siu.edu} To: {Microscopy-at-msa.microscopy.com} Sent: Monday, October 18, 2004 10:26 PM
Mike Yes, it is horses for courses. The astronomers up the road from here at Edinburgh Royal Observatory routinely scan 8" x 10" negatives at 8µm resolution, which gives huge digital images. (No way is the scanner made by Epson). I agree we don't all need that. But in a TEM, silver film is still the only straightforward way to have both widefield and high resolution, without stitching digital images together. Because the size and resolution of digital sensors is more limited one still has to choose whether to put the sensor in the 35mm port for widefield and lower resolution, or in the sheet-film position (or below) for high resoution with tunnel vision. Beyond that, I am not going to argue with you about the advantages of digital cameras. They're great, convenient, sometimes extremely sensitive, you get the image straight way, and much more. I use them myself (though not on the EM, alas). It is just that the performance envelope of the one does not yet coincide with the other.
I think you slightly mis-read my point about the binary nature of the silver image. Like newsprint, it is composed of black bits and white bits. My point is that as soon as we start talking of requiring high bit-depth to contain the grey-scale data we must be sampling at resolution that encompasses whole populations of grains, rather than single grains close to the spatial resolution limit of the film.
Chris Jeffree
----- Original Message ----- } From: "Mike Bode" {mb-at-soft-imaging.com} To: {microscopy-at-msa.microscopy.com} Cc: "'Chris Jeffree'" {c.jeffree-at-ed.ac.uk} Sent: Tuesday, October 19, 2004 5:56 PM
Jeff,
I think you expressed nicely what I wanted to say: "It is just that the performance envelope of the one does not yet coincide with the other." My point was that most people don't even need the film's high resolution AND field of view at the same time. You can trade off what you don't need in film for other parameters like linearity or convenience.
About the newspaper analogy: I only wanted to say that there is more information in a newspaper than you usually need on a daily basis. Period.
Regarding the binary nature of the film grains: I agree with you. If you only look at single grains, you talk about bits (1 or 0) of information, not bytes. To get to bytes you have to look at several grains, as you suggested.
Looks like we're saying the same thing ;-)
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: Chris Jeffree [mailto:c.jeffree-at-ed.ac.uk] Sent: Tuesday, October 19, 2004 12:08 To: Mike Bode Cc: microscopy-at-msa.microscopy.com
Hi, All-
A zoologist here wants to measure the depth of a couple of types of coral calices. I told him I wasn't enthusiastic about sectioning them and proposed breaking the coral up and looking for some fortuitous breaks for which he could measure the depth with the light microscope (they are appropriately sized for a stereo scope) or the SEM if he desired. But he is now enamored of the idea of something that can give the topography "like how they map the ocean floor". Would this be acoustic microscopy? Some other type of microscopy? Any other ideas? I have TEMs, SEMs, light and confocal microscopes at my disposal, but he would be happy to send it somewhere for this analysis.
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* ****************************************************************************
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 19 14:43:15 2004
I want to move slightly off the subject of the digital size equivalency of film. The point I want to make is about the number of pixels necessary to obtain specific resolution.
The concept of resolution defined as the ability to resolve two points (respectively two lines) is not very useful practical approach. Two pixels per line is hardly enough to properly image the size and shape of an object with an arbitrary shape.
Something in the range of 5 to 10 pixel per line is an adequate measure to obtained good resolution and true shape and size of objects.
This requirement is close to a lower limit of 5 for film where the crystal grains are randomly distributed. For a square net of digital pixel system a value close to 10 pixel per line would be necessary for proper imaging.
With film capable of resolving about 160 lines per mm we can get about 15 to 30 lines per mm PROPERLY RESOLVED on film.
Krassimir.
--------------------------------------------------------- Krassimir N. Bozhilov, PhD EM Scientist and Manager Central Facility for Advanced Microscopy and Microanalysis University of California Riverside, CA 92521
Tel 951 827 2998 Fax 951 827 2489 --------------------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 19 15:52:39 2004
I have no notion what a "coral calice" may be but I do use acoustic microscopes. There are two commercially available that I have used. The manufacturers are Sonoscan, Inc. and Sonix. Acoustic microscopy is somewhat different than optical or electron, etc. Depending on the spatial resolution you require this may be a good method of imaging. It also allows for sub-surface imaging similar to sonar in naval applications.
Some constraints may be the mechanical compliance of the material you are imaging. Mechanically stiffer materials image much better than say "squishy" biological specimens that are very compliant. You probably could get a great deal of help from the eqpt. vendors and they have some reasonably good application notes. If I recall, there may have been some articles in Microscopy Today on bio imaging using acoustics.
Hope this is of some help.
Regards,
Peter Tomic Agere Systems Allentown, PA
-----Original Message----- } From: Tina Carvalho [mailto:tina-at-pbrc.hawaii.edu] Sent: Tuesday, October 19, 2004 2:53 PM To: Microscopy Listserver
Hi, All-
A zoologist here wants to measure the depth of a couple of types of coral calices. I told him I wasn't enthusiastic about sectioning them and proposed breaking the coral up and looking for some fortuitous breaks for which he could measure the depth with the light microscope (they are appropriately sized for a stereo scope) or the SEM if he desired. But he is now enamored of the idea of something that can give the topography "like how they map the ocean floor". Would this be acoustic microscopy? Some other type of microscopy? Any other ideas? I have TEMs, SEMs, light and confocal microscopes at my disposal, but he would be happy to send it somewhere for this analysis.
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* ************************************************************************ ****
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 19 16:18:50 2004
I am an undergraduate student of Materials Science at Oxford University. In a Team Design Project which is part of our course, my team has the task to design a cyclic loading stage for use in a SEM (in situ) to enable observation of samples under load and the dynamic evolution of damage during cycling. The project does not only include the actual design but also a market evaluation. In order to get a feeling for the demand of such a stage I would like to ask for your expert advice on a few issues. I have designed a little questionnaire which should take less than five minutes to fill out. This would greatly benefit our Design Project. Needless to say we would be very grateful for your feedback.
On behalf of my team I would like to thank you very much in advance for helping us in estimating the demand for such a cyclic loading stage.
Best regards, Markus Mittermaier
Questionnaire: A Cyclic Loading Stage For Use in a Scanning Electron Microscope
1) What is your primary research area:
2) Have you ever heard of cyclic loading stages being used in SEM? If yes, please specify:
3) Do you encounter situations where a cyclic loading stage for a SEM would benefit your research? If yes, please specify:
4) Can you imagine using a cyclic loading stage of small dimensions for applications other than in a SEM? If yes, how?
5) What market price do you expect for such a cyclic loading stage which would have to be tailor-made (to a certain extent) or include an adapter to fit inside your microscope. Please comment on your expectation about the price (regardless of whether you would be willing to pay that amount).
A: 15000-20000 US Dollars B: 20000-25000 US Dollars C: 30000-35000 US Dollars D: more than 35000 US Dollars E: other:
6) Which actuation mechanism to supply the load to the sample in the SEM would you prefer. a) Hydraulic actuation b) Piezoelectric actuation c) Other:
7) What is your estimate about the demand for small scale cyclic loading stages in academia and industry worldwide?
Depending on the dimensions involved and the shape, you might want to try a light section microscope and do it non-destructively. These are used in industry to measure small height differences. The method is also contact-less, since it involves measuring the parallax offset of a tightly collimated line of light projected obliquely onto the surface. It can be used on wet paint or ink, for example. In principle, it could even be used on the soft tissue of live coral.
By using a step-repeat-step-repeat process one could also record a series of contour lines across the sample.
John Twilley
Tina Carvalho wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Hi, All- } } A zoologist here wants to measure the depth of a couple of types of coral } calices. I told him I wasn't enthusiastic about sectioning them and } proposed breaking the coral up and looking for some fortuitous breaks for } which he could measure the depth with the light microscope (they are } appropriately sized for a stereo scope) or the SEM if he desired. But he } is now enamored of the idea of something that can give the topography } "like how they map the ocean floor". Would this be acoustic microscopy? } Some other type of microscopy? Any other ideas? I have TEMs, SEMs, light } and confocal microscopes at my disposal, but he would be happy to send it } somewhere for this analysis. } } 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* } ****************************************************************************
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 19 17:54:31 2004
When comparing resolution of film with the resolution of a digital camera, you need to realize two things: 1) That film is not digital. It is analog, and its frequency response is described by a modulation transfer function. Here, for example, is the URL for the curve for Kodak T-Max black and white film: http://www.kodak.com/global/en/professional/support/techPubs/f4016/f002_ 0542ac.gif 2) 2) Pixel counts are not resolution. Resolution is the ability to resolve two features represented by intensity peaks. Generally, one pixel for bright and one pixel for dark (the Nyquist criterion-you all know that). Comparing digital to analog requires choosing the threshold for the analog resolution, and I am not sure what the critical criteria are for making that choice. For example, is T-max resolution 160 line pairs per millimeter (35% Response) or is it 60 line pairs per millimeter (90% response), or is it something in between?
John Mardinly Intel
-----Original Message----- } From: John J. Bozzola [mailto:bozzola-at-siu.edu] Sent: Monday, October 18, 2004 2:26 PM To: Microscopy-at-msa.microscopy.com
Could someone provide the digital equivalency (in terms of resolution) of 3.25 x 4 inch TEM film.
I have read that a 6 megapixel digital file has the resolution capability of a 35 mm grayscale negative. If this is true, then an 18 megapixel digital file would be equivalent to a TEM grayscale negative in terms of resolution capability.
I was thinking (based on darkroom enlargement capabilities of TEM films) that a 180 MP digital file would be more likely to exhibit the resolution one would see in a TEM negative.
Thanks for the information.
JB -- ############################################################## 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 Email: bozzola-at-siu.edu Web: http://www.siu.edu/~image/ ##############################################################
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 19 18:11:14 2004
You are correct in that a certain oversampling is necessary to be able to fully reconstruct a given signal. This was explored in detail by Shannon and Nyquist (I can't give references here, but a web search of Nyquist will result in many hits). In essence the result is that a given signal must be oversampled by a factor of 2 to allow a perfect reconstruction of the original signal. In other words: If your phosphor resolution is 5 microns (5 micron grains), you need to sample it with a pixel size of 2.5 microns. An oversampling of less than that might lead to aliasing (convolution of signals that cannot be corrected for), oversampling of much more than a factor of 2 will not increase the quality significantly.
However, the final result is also influenced by the Modulation Transfer Function of any lens or other optical element you might have between the phosphor and the CCD. So, the number of pixels is an important number, but if you couple a high-quality CCD with a mediocre lens, the result will be mediocre at best.
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: K.N. Bozhilov [mailto:bozhilov-at-ucr.edu] Sent: Tuesday, October 19, 2004 13:50 To: microscopy-at-msa.microscopy.com
Chris and all,
I want to move slightly off the subject of the digital size equivalency of film. The point I want to make is about the number of pixels necessary to obtain specific resolution.
The concept of resolution defined as the ability to resolve two points (respectively two lines) is not very useful practical approach. Two pixels per line is hardly enough to properly image the size and shape of an object with an arbitrary shape.
Something in the range of 5 to 10 pixel per line is an adequate measure to obtained good resolution and true shape and size of objects.
This requirement is close to a lower limit of 5 for film where the crystal grains are randomly distributed. For a square net of digital pixel system a value close to 10 pixel per line would be necessary for proper imaging.
With film capable of resolving about 160 lines per mm we can get about 15 to 30 lines per mm PROPERLY RESOLVED on film.
Krassimir.
--------------------------------------------------------- Krassimir N. Bozhilov, PhD EM Scientist and Manager Central Facility for Advanced Microscopy and Microanalysis University of California Riverside, CA 92521
Tel 951 827 2998 Fax 951 827 2489 --------------------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 19 18:25:30 2004
On Oct 19, 2004, at 12:50 PM, K.N. Bozhilov wrote:
} I want to move slightly off the subject of the digital size } equivalency of } film. The point I want to make is about the number of pixels necessary } to } obtain specific resolution. } } The concept of resolution defined as the ability to resolve two points } (respectively two lines) is not very useful practical approach. } Two pixels per line is hardly enough to properly image the size and } shape of } an object with an arbitrary shape. } } Something in the range of 5 to 10 pixel per line is an adequate } measure to } obtained good resolution and true shape and size of objects. } } This requirement is close to a lower limit of 5 for film where the } crystal } grains are randomly distributed. For a square net of digital pixel } system a } value close to 10 pixel per line would be necessary for proper imaging. } } With film capable of resolving about 160 lines per mm we can get about } 15 to } 30 lines per mm PROPERLY RESOLVED on film. } Dear Krassimir, If one considers the situation in reciprocal space, one has all the information out to a limiting spatial frequency, which is determined by the sampling; i.e., the pixel size. It is true that the Fourier transform of an image with continuous sampling--equivalent to unlimited spatial frequency--which is cut off at a particular resolution, is not exactly the same as the FFT of a pixelated image with a pixel size corresponding to the same resolution (twice the pixel size, as previously stated), but as one looks at features at a somewhat larger resolution, say 2/3 of the Nyquist frequency (or three pixels), the magnitudes and phases of the FFT are pretty close to those in the continuous-sampling case. 5 to 10 pixels per line is OK when you are trying to define the line, but higher-frequency information is available from fewer pixels, and, in particular, objects smaller than 5 pixels can be determined to be individual objects. I am confident that I can determine the structures present in a micrograph to a resolution of 2/3 Nyquist in the case that the micrograph has sufficient contrast and S/N ratio, both of which play a big role in the resolution one can obtain. If, for example, one has a pattern of infinitely dark lines separated by regions of 100% transmission, one can describe them with sub-pixel accuracy, since a pixel at 0.3 OD (=50% transmission) is half black, so the edge of the line must bisect the pixel. (Of course, the line must have finite thickness to be seen at all, and I am assuming that it is thicker than one pixel.) On the other hand, an image where the lines are at 1 OD and the spaces are at 0.95 OD, and where there is noise--a typical condition for frozen-hydrated biological specimens--I would expect it to be difficult to define lines with 5 to 10 pixels. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 19 18:59:41 2004
Thanks to everyone who sent responses to my question regarding the comparative resolution of TEM film versus digital imaging. Since I was giving a lecture on electron micrography and darkroom methods in TEM, I thought it would be informative for students to understand the advantages and disadvantages of film versus digital imaging. Resolution is always a concern, so it is informative to be able to compare the two technologies with data. We use both methods of imaging in our TEMs, depending on the needs of the research project.
-- ############################################################## 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 Email: bozzola-at-siu.edu Web: http://www.siu.edu/~image/ ##############################################################
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 19 22:09:21 2004
Wow! Lots of interesting replies to my query on measuring the depth of coral calices. Most of them asking what a coral calyx is and about how big it would be! For the non-invertebrate-zoology types on the List, the soft-bodied coral polyp (kind of upside-down jellyfish-like) secretes and lives in a calcium carbonate "skeleton" (lots of which make up the world's coral reefs). He only wants to measure the depth of some of the cups that the polyps live in; nothing about the tissue itself. Each cup (calyx) is probably about 0.5 - 1.5 mm deep, and about 2 mm across. I don't have an image handy, but I found one on the Web at http://www.uga.edu/caur/coral.jpg - it's a picture of a coral with a bunch of calices. Try to topomap that!
Those of you who attend Microscopy & Microanalysis 2005 here next year can put on a mask and snorkel and see for yourselves...
I have another project coming up where someone wants me to section deep-sea soft corals and see if there are bacteria inside. I already have found out they are not soft at all, but are armored to the max, inside and out! But that I can deal with. I hope.
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* ****************************************************************************
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 19 22:17:51 2004
I have been following this discussion with great interest. The cryo-TEM community has been interested in these questions for some time. The most recent comparison of which I am aware was published by Zhang et al., "Automated image acquisition and processing using a new generation of 4K x 4K CCD cameras for cryo-electron microscopic studies of macromolecular assemblies," Journal of Structural Biology, 143, 135-144 (2003). From the abstract: "We demonstrate that at 120 kV, and at a nominal magnification of 67,000X, power spectra and signal-to-noise ratios for the new 4K CCD camera are comparable to values obtained for film images scanned using a Zeiss scanner to resolutions as high as ~ 1/6.5 A." In the article, the authors noted that this camera still only had approximately one third of the area of the TEM negative. A close examination of Figure 4 in the article showed that the signal to noise ratio of film was noticeably better than the CCD at higher spatial frequencies.
As one who has used both film and 1K x 1K CCD cameras for TEM imaging for several years, I really appreciate the "instant gratification" that accompanies the CCD camera (especially for quantitative image analysis). I also appreciate the high resolution, large field of view available in a 3.25" x 4" TEM negative. Happily, I can choose whichever sensor (or both) that I need to solve a particular problem.
Standard disclaimer: my employer manufactures film, solid state image sensors, and scanners.
John Minter jrminter-at-rochester.rr.com
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 20 06:20:38 2004
take a look at the micro X-ray tomography instrument from Skyscan - http://www.skyscan.be/next/home.htm
At 08:32 PM 10/19/2004, Tina Carvalho wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 20 07:12:01 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (bwareham-at-utah.gov) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, October 18, 2004 at 17:01:52 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] Live cell stain
Question: Hi all, An associate of mine is looking for a live cell stain for cell culture. If anyone has any ideas, we would appreciate hearing them. Thank you, Beverly Wareham
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (lynda-at-biotech.ufl.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, October 19, 2004 at 07:56:09 ---------------------------------------------------------------------------
I am in need of learning if there is a way of lifting a paraffin stained section from a glass slide for EM. I have a PI that has a case he would like to have worked up by EM but the best material he has is an H&E stained section on a slide. Any ideas?
Mike, As a non-expert, doesn't Nyquist say the over-sampling must be greater than 2? I believe 2 can give considerable aliasing but the problem can be overcome at 2.1. Is this correct or did I miss something?
Ken Converse
owner QUALITY IMAGES Servicing Scanning Electron Microscopes Since 1981 16 Creek Rd. Delta, PA 17314 717-456-5491 Fax 717-456-7996 kenconverse-at-qualityimages.biz qualityimages.biz
-----Original Message----- } From: Mike Bode [mailto:mb-at-soft-imaging.com] Sent: Tuesday, October 19, 2004 7:17 PM To: 'K.N. Bozhilov' Cc: microscopy-at-msa.microscopy.com
Krassimir,
You are correct in that a certain oversampling is necessary to be able to fully reconstruct a given signal. This was explored in detail by Shannon and Nyquist (I can't give references here, but a web search of Nyquist will result in many hits). In essence the result is that a given signal must be oversampled by a factor of 2 to allow a perfect reconstruction of the original signal. In other words: If your phosphor resolution is 5 microns (5 micron grains), you need to sample it with a pixel size of 2.5 microns. An oversampling of less than that might lead to aliasing (convolution of signals that cannot be corrected for), oversampling of much more than a factor of 2 will not increase the quality significantly.
However, the final result is also influenced by the Modulation Transfer Function of any lens or other optical element you might have between the phosphor and the CCD. So, the number of pixels is an important number, but if you couple a high-quality CCD with a mediocre lens, the result will be mediocre at best.
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: K.N. Bozhilov [mailto:bozhilov-at-ucr.edu] Sent: Tuesday, October 19, 2004 13:50 To: microscopy-at-msa.microscopy.com
Chris and all,
I want to move slightly off the subject of the digital size equivalency of film. The point I want to make is about the number of pixels necessary to obtain specific resolution.
The concept of resolution defined as the ability to resolve two points (respectively two lines) is not very useful practical approach. Two pixels per line is hardly enough to properly image the size and shape of an object with an arbitrary shape.
Something in the range of 5 to 10 pixel per line is an adequate measure to obtained good resolution and true shape and size of objects.
This requirement is close to a lower limit of 5 for film where the crystal grains are randomly distributed. For a square net of digital pixel system a value close to 10 pixel per line would be necessary for proper imaging.
With film capable of resolving about 160 lines per mm we can get about 15 to 30 lines per mm PROPERLY RESOLVED on film.
Krassimir.
--------------------------------------------------------- Krassimir N. Bozhilov, PhD EM Scientist and Manager Central Facility for Advanced Microscopy and Microanalysis University of California Riverside, CA 92521
Tel 951 827 2998 Fax 951 827 2489 --------------------------------------------------------
___________________________________________________________ $0 Web Hosting with up to 120MB web space, 1000 MB Transfer 10 Personalized POP and Web E-mail Accounts, and much more. Signup at www.doteasy.com
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 20 08:40:01 2004
What you need to do is invert a beem capsule with epon on top of the section, polymerize, and then dip in liquid nitrogen - the block will pop off with section embedded. (Of course, remove the coverslip first!)
Peggy Sherwood Lab Associate, Photopathology Wellman Center for Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
-----Original Message----- } From: lynda-at-biotech.ufl.edu [mailto:lynda-at-biotech.ufl.edu] Sent: Wednesday, October 20, 2004 8:20 AM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (lynda-at-biotech.ufl.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, October 19, 2004 at 07:56:09 ---------------------------------------------------------------------------
I am in need of learning if there is a way of lifting a paraffin stained section from a glass slide for EM. I have a PI that has a case he would like to have worked up by EM but the best material he has is an H&E stained section on a slide. Any ideas?
I agree with Michael that the phosphate buffer is the factor contributing to the contamination you have observed. Next time use cacodylate buffer. Maunsbach and Afzelius book: "Biomedical Electron Microscopy, Illustrated methods and interpretations" comments on pages 76 & 77 on phosphate buffer precipitate which is not generally well understood. They later comment on the vulnerablity of lipid rich tissue (yours had myelin) to this artifact and how this only occurs with phosphate buffers. They reference an article from Stain Technology, 1979 by Ellis, E.A. & Anthony, D.W. vol. 54:282-285 which discusses how to remove this precipitate.
Hope this helps!
Karen Bentley
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
-----Original Message----- } From: BENJAMIN K AUGUST [mailto:bkaugust-at-facstaff.wisc.edu] Sent: Monday, October 18, 2004 5:24 PM To: microscopy-at-msa.microscopy.com; microscopy-at-msa.microscopy.com
Our lab has routinely processed samples according to the methods posted at http://www.micro.wisc.edu/smith in the past without artifacts, but recently we have encountered an unknown contaminant seen as an electron dense (opaque) amorphic precipitate. Please visit the web-site listed above for examples of the artifact and detailed processing methods. We would like your opinions as to the source of this contamination so we might avoid it in future experiments. Your comments are much appreciated.
Ben August U.W. Electron Microscopy Facility Medical School University of Wisconsin - Madison bkaugust-at-facstaff.wisc.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 20 09:06:21 2004
It's been a while since I dove into the details of the Nyquist theorem. The theorem is usually stated like this:
Nyquist's theorem: A theorem, developed by H. Nyquist, which states that an analog signal waveform may be uniquely reconstructed, without error, from samples taken at equal time intervals. The sampling rate must be equal to, or greater than, twice the highest frequency component in the analog signal.
The "equal to" would imply that a factor of 2 is sufficient.
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: Ken Converse [mailto:kenconverse-at-qualityimages.biz] Sent: Wednesday, October 20, 2004 07:00 To: 'Mike Bode' Cc: MSA Listserver
Krassimir,
You are correct in that a certain oversampling is necessary to be able to fully reconstruct a given signal. This was explored in detail by Shannon and Nyquist (I can't give references here, but a web search of Nyquist will result in many hits). In essence the result is that a given signal must be oversampled by a factor of 2 to allow a perfect reconstruction of the original signal. In other words: If your phosphor resolution is 5 microns (5 micron grains), you need to sample it with a pixel size of 2.5 microns. An oversampling of less than that might lead to aliasing (convolution of signals that cannot be corrected for), oversampling of much more than a factor of 2 will not increase the quality significantly.
However, the final result is also influenced by the Modulation Transfer Function of any lens or other optical element you might have between the phosphor and the CCD. So, the number of pixels is an important number, but if you couple a high-quality CCD with a mediocre lens, the result will be mediocre at best.
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: K.N. Bozhilov [mailto:bozhilov-at-ucr.edu] Sent: Tuesday, October 19, 2004 13:50 To: microscopy-at-msa.microscopy.com
Chris and all,
I want to move slightly off the subject of the digital size equivalency of film. The point I want to make is about the number of pixels necessary to obtain specific resolution.
The concept of resolution defined as the ability to resolve two points (respectively two lines) is not very useful practical approach. Two pixels per line is hardly enough to properly image the size and shape of an object with an arbitrary shape.
Something in the range of 5 to 10 pixel per line is an adequate measure to obtained good resolution and true shape and size of objects.
This requirement is close to a lower limit of 5 for film where the crystal grains are randomly distributed. For a square net of digital pixel system a value close to 10 pixel per line would be necessary for proper imaging.
With film capable of resolving about 160 lines per mm we can get about 15 to 30 lines per mm PROPERLY RESOLVED on film.
Krassimir.
--------------------------------------------------------- Krassimir N. Bozhilov, PhD EM Scientist and Manager Central Facility for Advanced Microscopy and Microanalysis University of California Riverside, CA 92521
Tel 951 827 2998 Fax 951 827 2489 --------------------------------------------------------
___________________________________________________________ $0 Web Hosting with up to 120MB web space, 1000 MB Transfer 10 Personalized POP and Web E-mail Accounts, and much more. Signup at www.doteasy.com
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 20 09:38:45 2004
I am trying to find information on physical cell sizes, such as diameter (volume) of the nucleus, etc., for various human cell types (cell lines) ? Are there on-line resources where this kind of general information is available ? I did not find this information on the ATCC website.
Regards,
Peter Van Osta
---------------------------------------------- Peter Van Osta, MD
} As a non-expert, doesn't Nyquist say the over-sampling must be } greater than 2? } I believe 2 can give considerable aliasing but the problem can be } overcome at 2.1.
2 is the Nyquist *limit*. Anything less than that and you are guaranteed to alias. At exactly 2 samples per cycle, you might get lucky or not get lucky. Consider sampling a sine wave at two samples per cycle. If you are lucky, you will pick the peaks of the sine wave and you will be able to reconstruct the wave perfectly (along with the a priori knowledge that it is a sine). But if you are unlucky, you will pick the zero crossings and you will reconstruct a zero frequency, zero amplitude wave.
If you sample at a higher frequency, then you can reconstruct the proper wave but, again, you must also add the a priori knowledge that it is a sine/cosine wave. The implication of the a priori knowledge is that if you have a wave that is more complex than a sine or cosine, you would need to do a Fourier decomposition of the wave to determine the highest non-negligible frequency component. Let's call that frequency F. You would then have to sample the wave at a frequency of at least 2F, preferably a little higher, to reconstruct the original wave properly.
Bruce Girrell
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 20 09:47:13 2004
One reason for (slight) oversampling is that if you sample at exactly 2x the maximum spatial frequency, you might by accident sample at the moment where the signal goes through all zero's.
---------------------------------------------- Peter Van Osta, MD
Director Imaging MAIA SCIENTIFIC Cipalstraat 3 B-2440 Geel, Belgium Tel.: +32 (0)14 570 620 Mobile: +32 (0)497 228 725 Fax.: +32 (0)14 570 621 Email: pvosta-at-maia-scientific.com Website: www.maia-scientific.com A Harvard Bioscience Company ---------------------------------------------- ======================================================== Ken Converse wrote: } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Mike, } As a non-expert, doesn't Nyquist say the over-sampling must be greater than } 2? I believe 2 can give considerable aliasing but the problem can be } overcome at 2.1. Is this correct or did I miss something? } } Ken Converse } } owner } QUALITY IMAGES } Servicing Scanning Electron Microscopes } Since 1981 } 16 Creek Rd. } Delta, PA 17314 } 717-456-5491 } Fax 717-456-7996 } kenconverse-at-qualityimages.biz } qualityimages.biz } }
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 20 10:07:37 2004
} Email: lynda-at-biotech.ufl.edu } Name: Lynda Schneider} } Organization: University of Florida } Question: } I am in need of learning if there is a way of lifting a paraffin stained } section from a glass slide for EM. I have a PI that has a case he would like } to have worked up by EM but the best material he has is an H&E stained } section on a slide. Any ideas? } } Thanks in advance. } Lynda Schneider --------------------------------------------------------------- Lynda, I have a protocol from the late 60's but never needed to try it. I do not know if it was published but the info. I have is as follows: Harvey Blank, MD and Carol Collins Dept. of Dermatology, Univ. of Miami
1. Clean slide and remove all stickers etc. 2. Slide into xylol several days to remove coverslip. 3. Slide coverslip without force off the slide. 4. Slide in fresh xylol to assure removal of all mounting medium. 5. Dip slide into 50% xylol + 50% propylene oxide (no time given) 6. Dip slide into 100% prophylene oxide 7. Dip slide into 50% prophylene oxide + 50% embedding medium (Epon type) 8. Invert slide, sections down, over a small dish filled with embedding medium (I would suggest using a mold or polypropylene plastic instead of glass and
make a few blank Beem Capsules to be used in step 13.) 9. Put into 60 degree C. oven over night to cure. 10.Remove slide from cured plastic while still warm. (They slide a fresh razor blade between the slide and the plastic. Some use liquid nitrogen to pop it off. Other comments welcome!) 11.Determine area of interest (LM) and scribe a box around it. 12.Cut out the area of interest. (Jeweler's saw works well or new razor blade) 13.Mount tissue side UP on a blank (with some reserved epoxy and put back into the oven 'til the next morning.)
I have found that if I put black Sharpie marker ink on the top of the blank and let it dry a bit before mounting the embedded tissue, it makes it much easier to align the block for sectioning.
I have heard that the result should be better than LM but not near as good as if the specimen was originally run up for TEM.
Lots of Luck, Pat Connelly Univ. of Pennsyvania psconnel-at-sas.upenn.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 20 10:17:58 2004
Well I have published several "pop-off" technique papers and will give you the references:
de Mesy Jensen, K.L.: "Pop-off" technique for FNA smears for diagnostic electron microscopy. Tech. Sample CY-1, American Society of Clinical Pathologists, 1987
de Mesy Jensen, K.L.: "Pop-off" technique for FNA smears for diagnostic electron microscopy. Tech. Sample CY-1, American Society of Clinical Pathologists, 1987
de Mesy Jensen, K.L., di Sant'Agnese, P.A.: Large block embedding and "pop-off" technique for immunoelectron microscopy with applications to prostatic endocrine-paracrine cells. Ultrastructural Pathology 16:51, 1992.
Here's the summary of what you should do:
You can look at an H & E section by "popping off" the section. All you have to do is remove the coverslip, rehydrate the slide from xylene to H2O and post fix in 1% OsO4 for 20 min., then dehydrate the slide (I use a plastic (5) slide mailer for processing) to 100% x 3 and infiltrate with epoxy resin. I always use Spurr resin for "popping off" a section. After the infiltration steps (usually 4 hrs. or overnight) you take a BEEM capsule with the cap removed, fill with liquid resin (up to top of capsule and slightly concave), quickly invert and place over the area you want to "pop-off" on the slide. Place the slide in the oven and polymerize overnight. The next day you can break the surface tension between the polymerized capsule and the glass by dipping the slide in liquid nitrogen (dip several times without submersing the capsule, just the base of the capsule attached to the slide) Wiggle the capsule and it should "pop-off", if it won't, then dip for several seconds more and try again.
Keep in mind the appearance of the "popped off" tissue will not look to good, since it was processed and stained onto a slide. Sometimes, it is better to take the paraffin block and core out the area of interest matched with the area of interest on the slide. Then I deparaffinize the tissue and post-fix in glut, OsO4 and process as if it were a normal EM specimen. Usually the tissue looks better than what you see on EM from a "popped off" tissue section.
Karen L. de Mesy Bentley, M.S.(previously Jensen) Associate Scientist & Project Manager Electron Microscope Research Core University of Rochester Medical Center Rochester, NY 14642 585-275-1954
-----Original Message----- } From: lynda-at-biotech.ufl.edu [mailto:lynda-at-biotech.ufl.edu] Sent: Wednesday, October 20, 2004 8:20 AM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (lynda-at-biotech.ufl.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, October 19, 2004 at 07:56:09 ---------------------------------------------------------------------------
I am in need of learning if there is a way of lifting a paraffin stained section from a glass slide for EM. I have a PI that has a case he would like to have worked up by EM but the best material he has is an H&E stained section on a slide. Any ideas?
sell several live fluorescent stains. I have used some of them on various cell types in culture, and they worked very well.
Lesley Weston.
on 20/10/2004 5:19 AM, by way of MicroscopyListserver at bwareham-at-utah.gov wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } 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 (bwareham-at-utah.gov) from } http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, } October 18, 2004 at 17:01:52 } --------------------------------------------------------------------------- } } Email: bwareham-at-utah.gov } Name: Beverly Wareham } } Organization: Utah Veterinary Diagnostic Laboratory } } Title-Subject: [Microscopy] [Filtered] Live cell stain } } Question: Hi all, } An associate of mine is looking for a live cell stain for cell culture. If } anyone has any ideas, we would appreciate hearing them. } Thank you, } Beverly Wareham } } --------------------------------------------------------------------------- }
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 20 11:03:45 2004
Be careful of any data you find. Cell of a given type vary in diameter, and thickness, depending on how they're prepared. Cultured cells spread out very thinly and so will be wider in diameter, and thinner, then, for example, cells collected in preservative, which will "round up" and be smaller in diameter. How much smaller depends on the type and concentration of preservative, and what happens to the cells next (e.g., applied to a slide [clear or dirty makes a difference]) and fixed, or applied to a slide and allowed to air-dry. Such details should be specified in conjunction with any cell size measurements.
For example, there is a 6-fold difference in area between mesothelial cells collected in 50% ethanol that are applied to a slide and wet-fixed, and fresh mesothelial cells that are applied to a slide and air-dried.
Gary Gill
-----Original Message----- } From: Peter Van Osta [mailto:pvosta-at-maia-scientific.com] Sent: Wednesday, October 20, 2004 9:46 AM To: MSA
Ken you are right, 2 is not enough - think about two pixels the same intensity next to each other and you'll see immediately that you can not distinguish a single two pixel by one pixel object from two adjacent single pixel objects. I believe that the number is something like 2.3 over sampling.
Bill
At 09:00 AM 10/20/2004, Ken Converse wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 20 12:46:27 2004
Greetings, A lower cost alternative to the designer probes if a simple viability test is needed is to use fluorescien diacetate and propidium iodide (available from Sigma). Live cells = green because an enzyme in the cell cleaves the membrane permeable fluorescien diacetate and dead cells exhibit red nuclei (after plasma membrane is compromised). See:
Jones and Senft. (1985). "An Improved Method to Determine Cell Viability by Simultaneous Staining with Fluorescien Diacetate-Propidium Iodide." J. Histochem. Cytochem. 33, 1, 77-79.
Regards, Karl
by way of MicroscopyListserver wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 20 13:10:11 2004
I have a glass-ceramic material, where glass and crystallites have the same composition. I want to perform the EBSD on crystallites of this material. I can not coat the material because I have to do diffraction. I used SE mode with 0.3 torr water pressure but the image quality is not good enough for 5 micron size crystallites which have the same composition as the glass and which are sticking out only few nanometer out of the glass surface (50-60 nm, i can make it 100-200 nm but etching but still having difficulty in finding out crystallites). I used the gas detector with 1 torr pressure (i can't go below that) and image quality improved but the diffraction pattern quality degraded to a level that I can not map it properly. Is there a simple way to do EBSD of these materials?
Sincerely Pradyumna Lehigh University
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 20 13:59:11 2004
We follow a similar procedure for lifting a paraffin section( unstained would produce better results). However,the procedure we use for processing tissue out of paraffin and sections does not reguire hydrating down to OsO4. We make up a 2%Osmium/xylene and start the processing at that point. It actually is shorter than starting with wet tissue,since the next step is into propelyne oxide,then resin mixtures.
Reference :
Kai Chien,R.L.Van de Velde and R.C. Heusser A One-Step Method for Re-embedding Paraffin Embedded Specimens for Electron Microscopy. EMSA 1982 pp356-357
Connie Gillies
-----Original Message----- } From: Bentley, Karen [mailto:Karen_Jensen-at-URMC.Rochester.edu] Sent: Wednesday, October 20, 2004 11:26 AM To: 'microscopy-at-msa.microscopy.com'
Lynda:
Well I have published several "pop-off" technique papers and will give you the references:
de Mesy Jensen, K.L.: "Pop-off" technique for FNA smears for diagnostic electron microscopy. Tech. Sample CY-1, American Society of Clinical Pathologists, 1987
de Mesy Jensen, K.L.: "Pop-off" technique for FNA smears for diagnostic electron microscopy. Tech. Sample CY-1, American Society of Clinical Pathologists, 1987
de Mesy Jensen, K.L., di Sant'Agnese, P.A.: Large block embedding and "pop-off" technique for immunoelectron microscopy with applications to prostatic endocrine-paracrine cells. Ultrastructural Pathology 16:51, 1992.
Here's the summary of what you should do:
You can look at an H & E section by "popping off" the section. All you have to do is remove the coverslip, rehydrate the slide from xylene to H2O and post fix in 1% OsO4 for 20 min., then dehydrate the slide (I use a plastic (5) slide mailer for processing) to 100% x 3 and infiltrate with epoxy resin. I always use Spurr resin for "popping off" a section. After the infiltration steps (usually 4 hrs. or overnight) you take a BEEM capsule with the cap removed, fill with liquid resin (up to top of capsule and slightly concave), quickly invert and place over the area you want to "pop-off" on the slide. Place the slide in the oven and polymerize overnight. The next day you can break the surface tension between the polymerized capsule and the glass by dipping the slide in liquid nitrogen (dip several times without submersing the capsule, just the base of the capsule attached to the slide) Wiggle the capsule and it should "pop-off", if it won't, then dip for several seconds more and try again.
Keep in mind the appearance of the "popped off" tissue will not look to good, since it was processed and stained onto a slide. Sometimes, it is better to take the paraffin block and core out the area of interest matched with the area of interest on the slide. Then I deparaffinize the tissue and post-fix in glut, OsO4 and process as if it were a normal EM specimen. Usually the tissue looks better than what you see on EM from a "popped off" tissue section.
Karen L. de Mesy Bentley, M.S.(previously Jensen) Associate Scientist & Project Manager Electron Microscope Research Core University of Rochester Medical Center Rochester, NY 14642 585-275-1954
-----Original Message----- } From: lynda-at-biotech.ufl.edu [mailto:lynda-at-biotech.ufl.edu] Sent: Wednesday, October 20, 2004 8:20 AM To: microscopy-at-microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (lynda-at-biotech.ufl.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, October 19, 2004 at 07:56:09 ------------------------------------------------------------------------ ---
I am in need of learning if there is a way of lifting a paraffin stained section from a glass slide for EM. I have a PI that has a case he would like to have worked up by EM but the best material he has is an H&E stained section on a slide. Any ideas?
An Anthropology student came into our EM facility with a sample I don't know how to handle. She wants to study the rate and extent of bacterial penetration into buried animal bones. She has a few previous papers showing methods for doing this, but the pictures are poor and methods are not always complete. Some people mention SEM, but don't show pictures. More often they seem to be using light microscopy of sections (30 microns) of resin embedded material.
We could do SEM but it isn't clear that this is the best approach, nor do we know which prep method to use if it is. Some groups critical point dry and use an SE detector. Others embed and use a backscatter detector. For LM we don't have any microtomes that will cut 30 micron sections and the histology group in another department has only cut decalcified material. The geology department has a diamond saw, but I don't know if this can be used on embedded material or whether we would get too much distortion if we try to cut or polish unembedded material.
Before we waste a lot of time trying all these options out, do any of you have experience with this type of work? Would she need to decalcify the bone? Should it be embedded? Would it be better to grind the surface and do SEM or cut sections and do light microscopy?
Thanks in advance for any suggestions you can offer.
Marie
Dr. Marie E. Cantino Director, Electron Microscopy Laboratory Associate Professor of Physiology and Neurobiology University of Connecticut Unit 3242 Storrs, CT 06269-3242 Phone: 860-486-3588 Fax: 860-486-6369
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 21 07:20:48 2004
I am looking for some information on the interaction of some popular cell culture compounds on image formation in digital (brightfield) microscopy.
Is there any information available on optical density (O.D.) spectroscopical analysis and turbity of Fetal Bovine Serum (FBS) and Fetal Calf serum (FCS) ? FCS causes a cell culture medium to become red and only partially transluscent instead of clear.
Phenol red (phenolsulfonphthalein, pH range 6.8 - 8.4) is a popular reagent to indicate pH changes in a cell culture, but it changes color from red to yellow with decreasing pH.
Are there measurements available to relate the concentration of phenol red to its capacity to absorb light (O.D.) and its influence on the overall color of cell cultures (spectroscopical analysis) ?
Regards,
Peter Van Osta
---------------------------------------------- Peter Van Osta, MD
I have a collaborator who is interested in looking at some plant material using the LM. He is interested in the staining of lignin and pectin using conventional techniques ( staining these with coloured stains where the components would stain different colours and could be distinguished from each other). My background is EM, and I am not set up for paraffin embedding, so it will have to be some sort of plastic embedding.
I know that there has been some discussion before on the use of LM stains on plastic sections. I'm wondering if the stains (such as Triarch quadruple stain) normally used for lignin and pectin and other plant materials would work if the plastic sections were etched or the plastic removed altogether. Or if there are other stains which I could use.
I hope to do some immuno work on these samples eventually, and maybe some TEM, but the LM has to be done first for us to decide whether to do the other techniques.
So, my questions are: 1. Can the specific botanical stains be used on plastic sections 2. Is there a preferred embedding plastic which I could use (LR white, methacrylate, other???)
Right now the samples are sitting in 2.5% glutaraldehyde in 0.05 M cacodylate buffer.
Thanks for any help with this.
Kind regards,
Paula.
Paula M. Allan-Wojtas Research Scientist - Food Microstructure/chercheur scientique - microstructure des aliments Food Safety and Quality team/ Salubrité et qualité des aliments Agriculture and Agri-Food Canada/Agriculture et Agroalimentaire Canada Telephone/Téléphone: 902-679-5566 Facsimile/Télécopieur: 902-679-2311 32 Main Street/ 32, rue Main Kentville, Nova Scotia/ Kentville (Nouvelle-Écosse) B4N 1J5
allanwojtasp-at-agr.gc.ca
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 21 08:20:19 2004
Assuming these bones aren't museum or type specimens that must remain unaltered ... We've done SEM of sub-perisosteal bacteria on bone, and superficial bacteria using the usual sort of glut fix/EtOH dehydration/CPD, and this works fine. For bacteria that penetrate bone, though ... if I had to do SEM, I'd use the standard ("standard") procedures, adding EtOH cryofracture in LN2 after the last 100% EtOH step to expose the interior. And, I'd try not doing this, but rather the usual methods and cracking the bone open after CPD. And then I won't accept it as definitive or the answer to the question, merely more information. You mostly likely will need to section decalcified bone to look for the bacteria, and your best resource is the collection of boneheads on histonet: List-Post: {mailto:histonet-at-lists.utsouthwestern.edu} List-Subscribe: {http://lists.utsouthwestern.edu/mailman/listinfo/histonet} , {mailto:histonet-request-at-lists.utsouthwestern.edu?subject=subscribe} List-Unsubscribe: {http://lists.utsouthwestern.edu/mailman/listinfo/histonet} , {mailto:histonet-request-at-lists.utsouthwestern.edu?subject=unsubscribe} List-Archive: {http://lists.utsouthwestern.edu/pipermail/histonet} List-Help: {mailto:histonet-request-at-lists.utsouthwestern.edu?subject=help}
There are several very good people well versed with bone histotechnique. I'd ask them.
Phil
} An Anthropology student came into our EM facility with a sample I } don't know how to handle. She wants to study the rate and extent of } bacterial penetration into buried animal bones. She has a few } previous papers showing methods for doing this, but the pictures are } poor and methods are not always complete. Some people mention SEM, } but don't show pictures. More often they seem to be using light } microscopy of sections (30 microns) of resin embedded material. } } We could do SEM but it isn't clear that this is the best approach, } nor do we know which prep method to use if it is. Some groups } critical point dry and use an SE detector. Others embed and use a } backscatter detector. For LM we don't have any microtomes that will } cut 30 micron sections and the histology group in another department } has only cut decalcified material. The geology department has a } diamond saw, but I don't know if this can be used on embedded } material or whether we would get too much distortion if we try to } cut or polish unembedded material. } } Before we waste a lot of time trying all these options out, do any } of you have experience with this type of work? Would she need to } decalcify the bone? Should it be embedded? Would it be better to } grind the surface and do SEM or cut sections and do light microscopy? } } Thanks in advance for any suggestions you can offer. } } Marie } } } Dr. Marie E. Cantino } Director, Electron Microscopy Laboratory } Associate Professor of Physiology and Neurobiology } University of Connecticut Unit 3242 } Storrs, CT 06269-3242 } Phone: 860-486-3588 } Fax: 860-486-6369
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 21 09:56:13 2004
Try Unicryl. It is a reliable dual-use EM/LM resin. LR White may work for you too.
usual disclaimers apply (to both). Chris
----- Original Message ----- } From: "Allan-Wojtas, Paula" {AllanWojtasP-at-AGR.GC.CA} To: {microscopy-at-msa.microscopy.com} Sent: Thursday, October 21, 2004 1:53 PM
Paula, Generally, stains that 'work' in paraffin also work in plastic. There is not even any need to etch. THe stains are small molecules and they penetrate easily.
To get information about specific stains, check the book by Steve Ruzin called Plant Micrototechnique and Microscopy, or Botanical Microtechnique and Cytochem by Berlyn and Miksche. Or if you can find it the book by O'Brian and McCully.
Having said that, if contrast is the only issue, its fine. But if you really want to know that the color is pectin, it is probably better to use antibody staining. There are quite a few good ones of those available. I think the dye stains for lignin are reliable, and I don't know of ab's for that.
For plant material and immuno, I have had very nice results with butyl-methyl-methacrylate, and I'd be happy to send you a protocol if you like (mainly for light level work).
As ever, Tobias
} } Hi, all, } } I have a collaborator who is interested in looking at some plant } material using the LM. He is interested in the staining of lignin } and pectin using conventional techniques ( staining these with } coloured stains where the components would stain different colours } and could be distinguished from each other). My background is EM, } and I am not set up for paraffin embedding, so it will have to be } some sort of plastic embedding. } } I know that there has been some discussion before on the use of LM } stains on plastic sections. I'm wondering if the stains (such as } Triarch quadruple stain) normally used for lignin and pectin and } other plant materials would work if the plastic sections were etched } or the plastic removed altogether. Or if there are other stains } which I could use. } } I hope to do some immuno work on these samples eventually, and maybe } some TEM, but the LM has to be done first for us to decide whether } to do the other techniques. } } So, my questions are: } 1. Can the specific botanical stains be used on plastic sections } 2. Is there a preferred embedding plastic which I could use (LR } white, methacrylate, other???) } } Right now the samples are sitting in 2.5% glutaraldehyde in 0.05 M } cacodylate buffer. } } Thanks for any help with this. } } Kind regards, } } Paula. } } Paula M. Allan-Wojtas } Research Scientist - Food Microstructure/chercheur scientique - } microstructure des aliments } Food Safety and Quality team/ Salubrité et qualité des aliments } Agriculture and Agri-Food Canada/Agriculture et Agroalimentaire Canada } Telephone/Téléphone: 902-679-5566 } Facsimile/Télécopieur: 902-679-2311 } 32 Main Street/ 32, rue Main } Kentville, Nova Scotia/ Kentville (Nouvelle-Écosse) } B4N 1J5 } } allanwojtasp-at-agr.gc.ca } } }
Hello: I am embedding small-grained materials using M-bond 610. I put the materials in Small capsule with M-bond, keeping ~70 C degree in an oven. However, it seems that M-bond is evaporated. I appreciate any suggestions you might have. The reason I am using M-bond is that thinning rate is low during ion-milling.
Thank you,
Hiromi Konishi Indiana University
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 21 12:32:04 2004
When doing EBSD with non-conductive small geologic particulates I've actually had the best using an extremely thin carbon coat and high vac rather than low vacuum. Never having measured it, my best guess is the coating is 5- 10nm - or as absolutely thin as you can get it, using a good vacuum (10-7 mbar range).
Just a suggestion.
} } I have following question: } } I have a glass-ceramic material, where glass and crystallites have the } same composition. I want to perform the EBSD on crystallites of this } material. I can not coat the material because I have to do diffraction. } I used SE mode with 0.3 torr water pressure but the image quality is not } good enough for 5 micron size crystallites which have the same } composition as the glass and which are sticking out only few nanometer } out of the glass surface (50-60 nm, i can make it 100-200 nm but etching } but still having difficulty in finding out crystallites). I used the gas } detector with 1 torr pressure (i can't go below that) and image quality } improved but the diffraction pattern quality degraded to a level that I } can not map it properly. Is there a simple way to do EBSD of these } materials? } } Sincerely } Pradyumna } Lehigh University } }
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Director 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
"RAM disk is NOT an installation procedure."
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 21 15:04:54 2004
Dear Pradymna, I would not etch the material at all, EBSD requires a very flat surface. The best is to go down all the steps of grinding and polishing to one micron diamond, then long polishing with 0.05 micron silica colloid (Syton). You may well find that the conditions for good EBSD are the opposite of the conditions for good imaging, you just change the conditions for each one. You will find that BSE imaging is best and works at any pressure. A colleague who did minerals said he polished, then did a very thin carbon coat, then polished again, then coated again, etc. This filled in any tiny cracks. A very thin carbon coat will not harm x-ray diffraction studies, it is invisible to the x-rays. Regards, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA Tel: 604-822-5648 Fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "Pradyumna Gupta" {png2-at-lehigh.edu} To: {Microscopy-at-MSA.Microscopy.Com} Sent: Wednesday, October 20, 2004 11:17 AM
Dear Hiromi, I understand that M-610 needs 100 to 120 degrees C to harden. It is usually used as a thin glue to stick pieces together for cross-sectioning, I do not know if it can be used as a mounting material. Good luck, Mary Mager Electron Microscopist Department of Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA Tel: 604-822-5648 Fax: 604-822-3619 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: {hkonishi-at-indiana.edu} To: {Microscopy-at-microscopy.com} Sent: Thursday, October 21, 2004 10:10 AM
Hello,
There is a possibility I might need to work with large gelatin capsules for inclusion/polymerization of hydrophilic resins (LR White or Unicryl).
However I will need a flat suppor for anatomic orientation.
I thought in cutting ACLAR film to provide flat support, as suggested here in a previous question about LR White and large samples embedding.
However I have some doubts, and since I have no experience I decided to post my doubts and ask for your advices.
-If I use large gelatin capsules as thos from Electron Microscopy Sciences (13 up to 22 mm diameter), I know I can cut a corresponding size ACLAR round fragment. I can then fill a little resin (to avoid oxygen bubble), put my ACLAR inside, then my sample and then fill with more resin, and then go to polymerization. Is this right?
-May I, perhaps cut one rounded ending of the gelatin capsule and glue it in an ACLAR sheet and then cut around, and then embed and polymerize? If this is factible, which glue can be used which seals properly and support either heat or cold UV polymerization? Cyanoacrilate(Super-Bonder, Loctite®)?
Thanks, Silvia
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From MicroscopyL-request-at-ns.microscopy.com Thu Oct 21 19:41:40 2004
As an avid collector/user of Bausch & Lomb metallographs at home and in my little business, I'd like to tell a story about amazing good luck ... and ask for a little help in finding a few things.
Years ago I happened upon some Leitz immersion objectives at a flea market. Many years later, I bumped into a buddy who was very interested in those objectives, but rather than give them away or try to fleece him, I suggested that he find and trade a B&L accessory that I had seen touted in the user's manual of the B&L Balphot I metallograph, the work horse of production metallography in the period during and after WWII. The accessory permits use of phase contrast in viewing opaque objects. My buddy searched for two years to no avail. I eventually traded for something else so he could acquire the Leitz objectives that he wanted. Fast forward another five years or so. My excellent microscope service person, John Sowers of Claymont, DE, came to my office for a regular service call with, to my astonishment, exactly one of those phase contrast accessories for the Balphot I. Fits perfectly, works like a charm. Now fast forward another five years or so. A B&L Research I metallograph appears on eBay, but in several disjointed pieces. I was lucky enough to get all the pieces plus a couple of giveaway parts from the seller. At about the same time, I found and purchased _another_ phase contrast accessory by B&L, very similar to the firat one but intended to fit a different 'scope. When it arrives, I discover to my amazement that it's the version that was intended to fit the Research metallograph. What luck ! But wait, that's not all. The Research I is missing the binocular eyepiece module, and I become resolved to making/adapting/fudging in order to gain easier access to those brilliant images. So I grab a pair of dimly illustrated binocular modules from eBay and discover to my delight that one is the missing Research I module and the other fits my Balphot I exactly as well. So I'm almost "there."
What's still missing is a 20X objective - the type with a tall, thin barrel and a wide base to accommodate the dark field illumination - and the parabolic reflectors that complete the dark field light path. Has anyone got surplus items in this category ? I'm not begging; I've been purchasing the other items out in the open market, so don't be bashful. I have all the other objectives that I'll ever need.
Word has it that those phase contrast accessories were never well accepted in the metallographic community, so extremely few were ever made. Anyone seen one in serious use out there ?
One more thing - the Research I metallograph had a weak method of attaching the two major portions of the main microscope body to each other - four tiny screws that easily strip their threads. I made and used successfully a jig to bore out the damaged threads, retap them, fit plugs with tapped holes the same size as the original screws, and machine flush the plugs. If anyone is in a similar dilemma, I'll gladly lend you the jig and associated tools. Nothing more than an eggbeater hand drill and a tap wrench are needed to complete the repair, once the jig is put to use.
Thanks, George Langford amenex-at-amenex.com
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 22 07:58:51 2004
There is an easier way if an eppendorf type tube is large enough for you. Check out this web page: http://www.biotech.ufl.edu/EM/data/disk.html
At 08:17 PM 10/21/2004 -0300, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program Scientific Director, Electron Microscopy P.O. Box 118525 217 Carr Hall University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295 fax- 352-846-0251
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 22 08:37:36 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rachel-wolf-at-uiowa.edu) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, October 21, 2004 at 14:54:23 ---------------------------------------------------------------------------
Question: Does anyone know of a seminar of some kind that gives training on Confocal microscopes?? We just got one at our lab and I will be the one using it, but I would like to have more education and background in using one before I jump all the way in. Thanks.
June 5-17, 2005 Lehigh University Bethlehem, PA USA
Courses include:
Introduction to SEM and EDS for the New SEM Operator Scanning Electron Microscopy and X-ray Microanalysis Problem Solving with Scanning Electron Microscopy Quantitative X-ray Microanalysis of Bulk Specimens and Particles Analytical Electron Microscopy Focused Ion Beam Instrumentation and Applications Particle and Fiber Characterization Scanning Probe Microscopy: from Fundamentals to Advanced Applications
For additional information please contact: -- ******************************** Sharon L. Coe Events Coordinator Lehigh Microscopy School 5 East Packer Avenue Bethlehem, PA 18015 610-758-5133 (phone) 610-758-4244 (fax) slc6-at-lehigh.ed www.lehigh.edu/microscopy
********************************
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 22 11:59:47 2004
I was wondering if anyone out there in ListServer land has had any experience with black and white laser printers used for printing electron micrographs.
Thank you
Ronald L. Austin Research Associate Dept. of Pathology LSU Medical Ct. Shreveport, LA. 71130 rausti-at-lsuhsc.edu 318-675-4557
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 22 18:08:53 2004
I was shown how RIP software can be used with the Epson 2200, 4000 and up model printers to print really great B&W prints. Anybody do that on this list? What are your thoughts and experience.
Damian Neuberger
Hello
I was wondering if anyone out there in ListServer land has had any experience with black and white laser printers used for printing electron micrographs.
Thank you
Ronald L. Austin Research Associate Dept. of Pathology LSU Medical Ct. Shreveport, LA. 71130 rausti-at-lsuhsc.edu 318-675-4557
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 23 07:46:25 2004
If you are interested in printing Ansel Adams quality EM prints a dedicated ink jet printer with Quad Black inks is the way to go - to get it right though requires RIP software..
Bill Miller
At 07:15 PM 10/22/2004, Damian Neuberger wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 23 09:20:24 2004
At 10:26 AM 10/23/2004, Chris Jeffree wrote: } Bill, Damian } } What is RIP software? } Chris } } Dr. Chris Jeffree } University of Edinburgh } } ----- Original Message ----- From: "Bill Miller" {microbill-at-mohawk.net} } To: "Damian Neuberger" {neuberger1234-at-comcast.net} ; "Austin, Ronald" } {RAusti-at-lsuhsc.edu} ; {microscopy-at-msa.microscopy.com} } Sent: Saturday, October 23, 2004 1:52 PM } Subject: [Microscopy] Re: RE: Laser printers } } } } } } } } ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 23 11:14:19 2004
We have a Zeiss 109 TEM that was very lightly used and needs to be moved. This microscope blew it's turbo pump a few years ago and has not been under vacuum since. So it is not working and I would not guarantee that it could be gotten up and running. However, it is a gold mine as to parts as it probably only has about 100-150 beam hours on it.
Free to academic home but you must arrange for transfer within 2 weeks. Small negotiable charge for commercial use. (i.e. Where parts are going to be resold).
Contact soon as it will be out the door in two weeks.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 23 12:21:55 2004
StudioPrint at http://www.ergosoftus.com/index.php
ImagePrint at http://www.colorbytesoftware.com/
} What is RIP software? } Chris } } Dr. Chris Jeffree } University of Edinburgh } } ----- Original Message ----- } } From: "Bill Miller" {microbill-at-mohawk.net} } To: "Damian Neuberger" {neuberger1234-at-comcast.net} ; "Austin, Ronald" } {RAusti-at-lsuhsc.edu} ; {microscopy-at-msa.microscopy.com} } Sent: Saturday, October 23, 2004 1:52 PM } Subject: [Microscopy] Re: RE: Laser printers } } } } } } } } ---------------------------------------------------------------------------- -} } - } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } ----------------------------------------------------------------------------- } } -- } } } } If you are interested in printing Ansel Adams quality EM prints a } } dedicated ink jet printer with Quad Black inks is the way to go - } } to get it right though requires RIP software.. } } } } Bill Miller } } } } At 07:15 PM 10/22/2004, Damian Neuberger wrote: } } } } } } } ---------------------------------------------------------------------------- } } } -- } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } } America } } } To Subscribe/Unsubscribe -- } } } http://www.msa.microscopy.com/MicroscopyListserver } } } On-Line Help } } } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } ---------------------------------------------------------------------------- } } } --- } } } } } } I was shown how RIP software can be used with the Epson 2200, 4000 } } } and up } } } model printers to print really great B&W prints. Anybody do that on } } } this } } } list? What are your thoughts and experience. } } } } } } Damian Neuberger } } } } } } Hello } } } } } } I was wondering if anyone out there in ListServer land has had any } } } experience with black and white laser printers used for printing } } } electron } } } micrographs. } } } } } } Thank you } } } } } } Ronald L. Austin } } } Research Associate } } } Dept. of Pathology } } } LSU Medical Ct. Shreveport, LA. 71130 } } } rausti-at-lsuhsc.edu } } } 318-675-4557 } } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 23 12:28:46 2004
Nope, RIP is not a "Mac thing" but a more general combination of hardware and software which a printer uses to convert vectorized objects text, lines, objects, etc... into the bitmaps (i.e. array of dots) which are printable using " printing" technologies that a modern printers employ.
All high end printers have RIP's built into their processors. The low end printers require your local CPU to convert objects into bitmaps which are then transferred to the printhead.
Postscript fonts and vector graphics for example have to be rasterized to be printed.
While images (BMP, TIFF, GIF, JPEG) are essentially already bitmaps and do not have to be processed (as much) to be output.
Nestor Your Friendly Neighborhood SysOp.
} X-Authentication-Warning: ns.microscopy.com: mail set sender to MicroscopyL-request-at-ns.microscopy.com using -f } From: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} } To: "Bill Miller" {microbill-at-mohawk.net} } Cc: {microscopy-at-msa.microscopy.com} } Subject: [Microscopy] Re: Re: RE: Laser printers } Date: Sat, 23 Oct 2004 17:20:28 +0100 } X-Priority: 3 } Status: } } } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 23 12:44:27 2004
-----Original Message----- } From: Chris Jeffree [mailto:c.jeffree-at-ed.ac.uk] Sent: Saturday, October 23, 2004 11:20 AM To: Bill Miller Cc: microscopy-at-msa.microscopy.com
OK, sounds like this may be a MAC thing. Chris
----- Original Message ----- } From: "Bill Miller" {microbill-at-mohawk.net} To: "Chris Jeffree" {c.jeffree-at-ed.ac.uk} Cc: {microscopy-at-msa.microscopy.com} Sent: Saturday, October 23, 2004 4:04 PM
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 23 13:36:05 2004
When I first took over Microscopy Today and sent my Adobe InDesign files that contained the "camera ready" version of the current MT issue to the printer, the printer prepress contractor said they would RIP it. I was upset naturally, given all the work I had put into it.
I since learned that RIP is a hardware-software combination that converts a vector image into a bit-mapped image. All PostScript printers contain a RIP that converts the PostScript commands into bit-mapped pages that the printer can output. The RIP is at the heart of the digital imaging process and does all the work. The ultimate result is a printed or imaged page, panel, sheet of film, or four half-tone screens for each of the printing press print stations (CMYK). In MT's case it is the four screens that are loaded into the printing press the prints MT. Hence the quotes around "camera ready", above. There is no camera in this century.
Ron Anderson
-----Original Message----- } From: Bill Miller [mailto:microbill-at-mohawk.net] Sent: Saturday, October 23, 2004 11:04 AM To: Chris Jeffree Cc: microscopy-at-msa.microscopy.com
RIPs (Raster Image Processors)
At 10:26 AM 10/23/2004, Chris Jeffree wrote: } Bill, Damian } } What is RIP software? } Chris } } Dr. Chris Jeffree } University of Edinburgh } } ----- Original Message ----- From: "Bill Miller" {microbill-at-mohawk.net} } To: "Damian Neuberger" {neuberger1234-at-comcast.net} ; "Austin, Ronald" } {RAusti-at-lsuhsc.edu} ; {microscopy-at-msa.microscopy.com} } Sent: Saturday, October 23, 2004 1:52 PM } Subject: [Microscopy] Re: RE: Laser printers } } } } } } } } -------------------------------------------------------------------------- ---- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 23 15:23:06 2004
Nestor OK, I'll admit I was just being gratuitously provocative, but we're still no nearer having an answer to the question arising from Bill & Damian's comments about RIPs: namely if (as implied) there is an advantage in using RIP software compared with printing the image direct from Adobe Photoshop using the standard Epson printer driver then why does that arise?
Best wishes Chris Jeffree
----- Original Message ----- } From: "Nestor J. Zaluzec" {zaluzec-at-microscopy.com} To: {microscopy-at-microscopy.com} Sent: Saturday, October 23, 2004 6:36 PM
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 23 16:22:05 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jmo12505-at-yahoo.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Saturday, October 23, 2004 at 15:06:00 ---------------------------------------------------------------------------
Email: jmo12505-at-yahoo.com Name: Judy
Organization: Saint Louis University
Title-Subject: [Microscopy] [Filtered] Advice on plastic options
Question: I have been using Epon Araldite plastic for many years for both EM and LM (1 micron) sectioning. I am gearing up for a new project that will involve embedding and 1 micron sectioning of a fairly large number of mouse brains. I will not need to do any EM on this tissue. I also have several undergraduates who are interested in participating in the project but have no prior histology experience. Are any of the newer plastics easier to work with for embedding and sectioning? Thanks in advance, -Judy
SEM WORKSHOP Presented by: Midwest Microscopy and Microanalysis Society
Affiliate of the Microscopy Society of America and the Microbeam Analysis Society of America
November 19th, 2004 8:00AM - 5:00PM Baxter Corporate Headquarters Deerfield, IL Directions and map at http://www.microscopy.org/MSALAS/MMMS
Registration Fees: MMMS members: before Nov. 5th - $ 25.00 after Nov. 5th - $ 35.00 Non-members: before Nov. 5th - $35.00, after Nov 5th - $ 45.00 (MMMS membership included in fee) Students: before Nov. 5th - $15.00, after Nov 5th - $ 25.00 (MMMS Student membership $5)
Vendors: We welcome vendors, tables for literature & exhibits available. Contact us for details.
8:00AM - 8:45AM Setup and registration
8:45 - 10:15AM - James Pawley, University of Wisconsin, What FE does for SEM?
10:15 - 10:45AM Break
10:45AM - 12:15PM - John Mackenzie, North Carolina State University, Work flow for digital imaging in a modern microscopy laboratory II
12:15PM - 1:15PM LUNCH - Included
1:15PM - 2:45PM - Alwyn Eades, Lehigh University, Crystallographic analysis in the Scanning Electron Microscope
2:45 - 3:15PM Break
3:15 - 4:45PM - Dennis Ward, Federal Bureau of Investigation, Microanalysis - A New Tool in Combating Terrorism
Please RSVP via email, phone, or fax to: Arvid Casler - MMMS Program Coordinator 847-566-7716 Voice Mail and Fax arvid_casler-at-fmo.com
NOTE: RSVPs will be responsible for registration fees
Robert Mierzwa Midwest Regional Sales Manager JEOL USA, Inc. 3906 Lisa Ave. Sheboygan WI 53083 TEL (920) 803-8945 FAX (920) 803-8946 Email: mierzwa-at-jeol.com
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 25 09:56:21 2004
Hi Debby, I sent your note to a friend who works on Zeiss scopes and has parts for the older orange-column scopes. He says he can have your 109 resurrected for a few thousand dollars. Are you sure you want to get rid of this scope? I will be happy to share service info with you. best, Beth
On Saturday, October 23, 2004, at 12:38 PM, Debby Sherman wrote:
} } } ----------------------------------------------------------------------- } ------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } -------- } } We have a Zeiss 109 TEM that was very lightly used and needs to be } moved. } This microscope blew it's turbo pump a few years ago and has not been } under } vacuum since. So it is not working and I would not guarantee that it } could } be gotten up and running. However, it is a gold mine as to parts as it } probably only has about 100-150 beam hours on it. } } Free to academic home but you must arrange for transfer within 2 weeks. } Small negotiable charge for commercial use. (i.e. Where parts are } going to } be resold). } } Contact soon as it will be out the door in two weeks. } } Debby } } Debby Sherman, Manager Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } S-052 Whistler Building } 170 S. University Street } West Lafayette, IN 47907 } http://www3.agriculture.purdue.edu/microscopy } } } } ********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) *******************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
I have a question about how to prepare SEM sample of automobile catalyst with metallic substrate. For the catalyst on ceramic substrate we drill a core, embed it in Epoxy and polish it. It is very hard to get a core from metallic substrate but still keep the original washcoat distribution without shaking it off by the strong vibration during cutting. Is there any special tool for cutting? After getting a core do I need to embed it in Epoxy and polish it?
Could you please kindly share your experience with me?
Note: The information contained in this message may be privileged and confidential and thus protected from disclosure. If the reader of this message is not the intended recipient, or an employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any dissemination, distribution or copying of this communication is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer. Thank you.
Wise ones in netland, This is a little off straight up microscopy but not all that far. My colleague Louis Cardenes asks:
I want to know how to make good microinjection needles with a Flaming/Brown micropipette Puller from Sutter Instrument. The needles are desired to be around 0.2 micrometer (internal diameter) and will be used for plant cell injections. What I need is a good protocol and the right platinum filament that should be used.
Can anyone help? Even to the point of suggesting an electrophys email listserver? Please include Louis' address (luisc-at-ibt.unam.mx) in your reply because he doesn't subscribe to this list.
Thanks all, Tobias -- _ ____ __ ____ / \ / / \ / \ \ Tobias I. Baskin / / / / \ \ \ Biology Department /_ / __ /__ \ \ \__ 611 N. Pleasant St. / / / \ \ \ University of Massachusetts / / / \ \ \ Amherst, MA, 01003 / / ___ / \ \__/ \ ____ http://www.bio.umass.edu/biology/baskin/ Voice: 413 - 545 - 1533 Fax: 413 - 545 - 3243
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 25 17:20:01 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (m.serracino-at-igag.cnr.it) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, October 25, 2004 at 08:55:29 ---------------------------------------------------------------------------
Question: Our Link eXL computer graphics board c1907 has died and we are scrambling for a quick replacement. Does anyone have a graphics board c1907 or have any not very expensive suggestions? Thanks to all who responded to the crisis of repairing or replacing the board.
Marcello Serracino m.serracino-at-igag.cnr.it Sx50 microprobe operator Institute of Environmental Geology and Geoengineering
I did read some note about the external connection of JEOL Sem 840 with PC.
There is a person from Alcoa Technical center proposed to use the MACADios card. It is able to interface the SEM and PC to grab SEM images. Does anyone has the contact and fax of the GW instruments who selling MAC ADios IIJr ? Let me know in your next email.
Thanks Kok Swee
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 26 07:20:05 2004
Thomas, depending on what kind of resolution you need and types of samples, you might try Profilometry. Zygo Corp. may be able to help you and they're right here in CT. The link is: www.zygo.com. Contact them of feel free to contact myself if you have any questions.
Ed Komarnicki Sr. Analytical Chemist MacDermid Inc. (203) 575-5750
Disclaimer: I have no capital interest in this company.
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello everyone,
I am currently involved in a research project that is using many different imaging modalities to look at objects of a variety of sizes. I am interested in using texture parameters to try to classify this material and was wondering if there was any existing computer programs or relevant papers written that could help my with my project.
Please feel free to email me if you think you can help. Thank you all for your time.
Thomas Sadowski Southern Connecticut State University
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 26 09:17:31 2004
A research associate is sought to serve as principle investigator and primary operator of the Digital Instruments Scanning Probe Microscope which is operated as a University user and outreach facility. The individual in this position will conduct research on materials characterization and behavior with the SPM and interact with faculty and students as well as industrial and governmental personnel. Materials and processing research involving composite materials, metals, ceramics, polymers, cement, asphalt, food, packaging materials, soils, and plants is being conducted with the SPM. In particular this unit is equipped with stages capable of varying temperature, ambient environment, tapping mode capabilities, scanning tunneling head, and indentation modules as part of the research studies on these materials. The person occupying this position will be responsible for the operation and maintenance of the unit, provide training of faculty, staff, graduate students and off-campus users and conduct research with this unit. In addition this person will be expected to develop contacts with potential users from off campus in the local and state-wide community to assist in generating research funds as part of the outreach activity of the SPM facility.
Qualifications: A Ph.D. in Materials Science, Physics, Chemistry, or Engineering is required. Experience with scanning probe microscopy required, with preferred experience with Digital Instruments Nanoscope series of controllers. The applicant must exhibit very high levels of English oral and written communication skills.
Applications: Position is open until filled. Salary is dependent on qualifications and experience. Send a complete curriculum vitae, graduate level transcript and three references to Michael J. Rich, Michigan State University, Composite Materials and Structures Center, 2100 Engineering Building, East Lansing, MI 48824-1226, ; Email: RICH-at-EGR.MSU.EDU
Additional information describing the Composite Materials and Structures Center facility is available at: http://www.egr.msu.edu/cmsc
MSU is an affirmative action, equal opportunity institution.
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 26 10:03:31 2004
I cannot unfortunately help Marcello Serracino with his eXL graphics board woes but I have the following eXL PC boards available (FREE) to whomever could use them from a circa 1991 Oxford/Link eXL microanalysis system which I decommissioned only yesterday.
PC2300 M.D.I (Mk.2) BD PC1789 MUX BD PC2733 GSP BD PC2391 Electron Signal Cond. BD PC1585 M.D.I Bd PC 1171-V1 W.D. Interface BD.
Nestor Your Friendly Neighborhood SysOp -- =========================================== Dr. Nestor J. Zaluzec Argonne National Lab Materials Science Division/Bldg 212 9700 S. Cass Ave Argonne, Illinois 60439 USA Tel: 630-252-7901, Fax: 630-252-4798 Email: Zaluzec-at-aaem.amc.anl.gov =========================================== TPMLab: http://tpm.amc.anl.gov NEESLab: http://neestpm.mcs.anl.gov MMSite: http://www.amc.anl.gov ===========================================
The box said ... "This program requires Win 95/98/NT or better..." So I bought a G3 Mac !
===========================================
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 26 10:14:09 2004
Marcello, in the another old box of electronic components hidden away in a corner I found a complete eXL GDP PC 1907 graphics board. !
I had forgotten that I replaced the eXL video system with a more modern display board which is VGA compatible. The 1907 board only works with the old style eXL monitors.
It hasn't been used in years, however I will mail it to you, if you still need it. Just contact me off line.
Nestor
Your Friendly Neighborhood SysOp
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (m.serracino-at-igag.cnr.it) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, October 25, 2004 at 08:55:29 ---------------------------------------------------------------------------
Question: Our Link eXL computer graphics board c1907 has died and we are scrambling for a quick replacement. Does anyone have a graphics board c1907 or have any not very expensive suggestions? Thanks to all who responded to the crisis of repairing or replacing the board.
Marcello Serracino m.serracino-at-igag.cnr.it Sx50 microprobe operator Institute of Environmental Geology and Geoengineering
--------------------------------------------------------------------------- -- =========================================== Dr. Nestor J. Zaluzec Argonne National Lab Materials Science Division/Bldg 212 9700 S. Cass Ave Argonne, Illinois 60439 USA Tel: 630-252-7901, Fax: 630-252-4798 Email: Zaluzec-at-aaem.amc.anl.gov =========================================== TPMLab: http://tpm.amc.anl.gov NEESLab: http://neestpm.mcs.anl.gov MMSite: http://www.amc.anl.gov ===========================================
The box said ... "This program requires Win 95/98/NT or better..." So I bought a G3 Mac !
===========================================
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 26 12:21:13 2004
Are single wall and multi-wall nanotube probes readily and commercially available for AFM? for STM?
-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)
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 26 15:40:11 2004
We are in need of Si (111) TEM sample (3mm disk) for our JEM-2010F CBED pattern test urgently. If you happen to have it, could you please mail me one sample as soon as possible? If you still need this sample, I will return it after our test. We appreciate your timely help.
Following is my mailing address: Qi Zhang 164 Phillips Hall, CB#3255 Department of Physics and Astronomy the University of North Carolina at Chapel Hill Chapel Hill, NC 27599-3255.
Thank you very much!
Regards, ************************************************** Qi Zhang, Ph.D 164 Phillips Hall, CB#3255 Department of Physics and Astronomy the University of North Carolina at Chapel Hill Chapel Hill, NC 27599 Tel: 919-843-2407 (Office) 919-843-0974 (EM Lab) Fax: 919-962-0480 E-mail: qizhang-at-physics.unc.edu
__________________________________ Do you Yahoo!? Yahoo! Mail - Helps protect you from nasty viruses. http://promotions.yahoo.com/new_mail
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 26 16:46:02 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (qizhang-at-physics.unc.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, October 26, 2004 at 16:34:14 ---------------------------------------------------------------------------
Email: qizhang-at-physics.unc.edu Name: Qi Zhang
Title-Subject: [Microscopy] [Filtered] MListserver: need Si [111] TEM sample (3mm disk)
Question: Dear colleagues,
We are in great need of Si [111] TEM sample (3mm disk) for our JEM-2010F CBED pattern test urgently. If you happen to have it, could you please mail me one sample as soon as possible? If you still need this sample, I will return it after our test. We appreciate your timely help.
Following is my mailing address: Qi Zhang 164 Phillips Hall, CB#3255 Department of Physics and Astronomy the University of North Carolina at Chapel Hill Chapel Hill, NC 27599-3255.
Thank you very much!
Regards, ************************************************** Qi Zhang, Ph.D 164 Phillips Hall, CB#3255 Department of Physics and Astronomy the University of North Carolina at Chapel Hill Chapel Hill, NC 27599 Tel: 919-843-2407 (Office) 919-843-0974 (EM Lab) Fax: 919-962-0480 E-mail: qizhang-at-physics.unc.edu
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (spradhan-at-siu.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, October 26, 2004 at 14:55:46 ---------------------------------------------------------------------------
Email: spradhan-at-siu.edu Name: sailesh
Organization: Southern Illinois University
Title-Subject: [Microscopy] [Filtered] MListserver: A used stage for the SEM
Question: hello everyone!
i am looking to see if there are any used loading stages for the hitachi S-2460N SEM. i am working on a project with a professor and we want to take some images of a thin-film under loading using the SEM(we want to work on speckle pattern interferometry with electron microscopy). what we have in mind is a stage(a door and a pass-through) that is designed for such purposes and can enable us to perform the tests on the SEM without having to go through the entire process of building it from scratch. so it would be nice to know of any such stages that are available. i would appreciate it very much if you could even give me a rough estimate of how much anything like it would cost. thank you...i shall look forward to hearing from you soon.
When I called to order chemicals for our AGFA Rapidoprint Processor (AGFA Rapidoprint Activator G182B and AGFA Rapidoprint Fixer QSO4A), I was told by my supplier that AGFA is no longer manufacturing them. I called the AGFA technical support line, but they could not suggest direct replacements nor would they share the formulas for the discontinued products with me so that I can make them myself. The ingredients in the two are common photo chemicals (for the activator water, ammonium thiosulfate and sodium sulfite; for the fixer water, sodium hydroxide and sodium sulfite).
Has anyone already grappled with this problem and found suitable replacements? I could find nothing on the web and the compositions of the products are not given in the Morgan & Morgan Photo Lab Index - at least not in the vintage copy in our library.
Least you think we're in the dark ages here, we do have a film scanner. There are just some old fashioned microscopists here (mainly me!) who still prefer stacks of study prints for data analysis over stacks of CDs.
Thanks for any forthcoming solutions ;)
Heather Owen
Dr. Heather A. Owen, Director Electron Microscope Laboratory Department of Biological Sciences University of Wisconsin - Milwaukee Lapham Hall, P.O. Box 413 Milwaukee, WI 53210 USA
Phone: (414)229-6816
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 27 01:17:13 2004
Afraid I haven't found the specific formulas, and not terribly familiar with machine processing, but have some thoughts that I hope will be helpful.
First, to mention that whoever told you of the chemistry you described has things reversed. Sodium hydroxide is a common high-energy alkalai "accelerator" used in some developers, while ammonium thiocyanate has long been a standard replacement for sodium thiosulfate in rapid fixers. I think rapidoprint is a paper that has the developing agent as well as restrainer built-in, and so the "developer" itself may be no more than the NaOH-sulfite solution, and if so it might just be a question of playing around with solution percentages. And in a very old copy of P.L.I. (1946) I find the following: "A 15-20% solution of ammonium thiosulfate is capable of more rapid fixation than a 35-40% solution of sodium hydroxide."
My own guess is that the one risk with experimenting with chemistry in the machine would be related to burning of the seals and possibly filters, but I find it hard to imagine any other ingredients than those you mentioned could be more corrosive than the hydroxide.
There's a site on the web called the Analog Photography Users Group, which has a forum dealing with traditional processes, and some of the members might be familiar with machine chemistry. Give it a try:
http://www.apug.org/forums/home.php
Finally, me being an old and "old fashioned" photographer, I'd encourage you: never, NEVER, apologize for using silver processes!!!!! (ad infinitum !!!!!'s)
Cheers, hope this helps, Larry Weissmann Tweed, Ontario, Canada Tel: (613) 478-4743
********************************** HEATHER OWEN WROTE
Hi All,
When I called to order chemicals for our AGFA Rapidoprint Processor (AGFA Rapidoprint Activator G182B and AGFA Rapidoprint Fixer QSO4A), I was told by my supplier that AGFA is no longer manufacturing them. I called the AGFA technical support line, but they could not suggest direct replacements nor would they share the formulas for the discontinued products with me so that I can make them myself. The ingredients in the two are common photo chemicals (for the activator water, ammonium thiosulfate and sodium sulfite; for the fixer water, sodium hydroxide and sodium sulfite).
Has anyone already grappled with this problem and found suitable replacements? I could find nothing on the web and the compositions of the products are not given in the Morgan & Morgan Photo Lab Index - at least not in the vintage copy in our library.
Least you think we're in the dark ages here, we do have a film scanner. There are just some old fashioned microscopists here (mainly me!) who still prefer stacks of study prints for data analysis over stacks of CDs.
Thanks for any forthcoming solutions ;)
Heather Owen
Dr. Heather A. Owen, Director Electron Microscope Laboratory Department of Biological Sciences University of Wisconsin - Milwaukee Lapham Hall, P.O. Box 413 Milwaukee, WI 53210 USA Phone: (414)229-6816
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 27 01:32:06 2004
I see that MACO Photo Products list processor chemistry, but I think this could be a conventional developer formulation and not a Rapidoprint-compatible activator. Anyway, it could be worth asking them your question?? Also, check out the other silver photography products they supply: http://www.mahn.net/Frameset.htm MACO is a division of Hans O. Mahn & Co. KG, Brookstieg 4, 22145 Stapelfeld, Germany Hotline: 040-237 008-88 von 9:00 - 16:00Uhr, e-mail: PHOTO-at-mahn.net
Agfa applied for a patent 1997 to extend the Rapidoprint principle (crudely put, the developer chemistry is built-in to the emulsion) to radiographic emulsions. In this application (which is available to view on the internet) they quote activator and fixer formulae as follows:
Composition of the activation liquid (per litre).
potassium hydroxide 30 g potassium sulphite 50 g potassium bromide 2 g ethylene diamine tetra acetic acid, sodium salt 1.5 g
Composition of the fixing liquid (per litre).
ammonium thiosulphate 100 g sodium sulphite 17 g sodium acetate 15 g citric acid 2.5 g acetic acid 13 ml
The rinsing liquid was distilled water.
I would experiment with 3% KOH to activate and your regular ammonium thiosulphate fixer at appropriate dilution to fix.
Chris Jeffree University of Edinburgh
----- Original Message ----- } From: "Heather A Owen" {owenha-at-csd.uwm.edu} To: {microscopy-at-msa.microscopy.com} Sent: Wednesday, October 27, 2004 12:17 AM
Dear All,
does anyone know the bethe range and Kanaya Okayama range of carbon from 20kev to 30kev. Let me know.
Many thanks Ks
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 27 08:07:54 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ajenning-at-cyllene.uwa.edu.au) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, October 26, 2004 at 22:58:40 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] IHC and TEM of zinc-fixed tissue
Question: I was wondering if anyone has used zinc fixative (based on Zinc salts in TBS and avaailable commercially for LM-level fixation) for EM preparation. It is supposed to retain many antigens which are masked by formalin, para or glutaraldehyde, also reducing the need for antigen retrieval. So far I haven't seen any reference including ultrastructural performance, nor has there been any satisfactory explanation of how the fixative actually works. As well as 3micron paraffin immunohistochemistry, I am interested in 1 micron araldite immunostaining. Many thanks Alison
Qi, A very simple way to make a Si(111) sample is to take a small piece of Si wafer and crush it in a mortar and pestle with a bit a ethanol. Then place a drop of the slurry on a holey carbon grid (you may have to play with the concentration a bit). You will need to hunt around a bit to get the correct zone axis, but you should be able to find nearly zone with out tilting too far. Look for small thin flakes that look nearly invisible with the TEM in focus.
Good luck, Ray
Ray D. Twesten, PhD. Center for Microanalysis of Materials Seitz Materials Research Laboratory 104 S. Goodwin Ave., Urbana, IL 61801 +1 217 244-6177 (Fax -2278)
-----Original Message----- } From: Zhang Qi [mailto:qzhangtj-at-yahoo.com] Sent: Tuesday, October 26, 2004 3:49 PM To: MSA listserver Cc: Lu-Chang Qin; Gary T Griego
Dear colleagues,
We are in need of Si (111) TEM sample (3mm disk) for our JEM-2010F CBED pattern test urgently. If you happen to have it, could you please mail me one sample as soon as possible? If you still need this sample, I will return it after our test. We appreciate your timely help.
Following is my mailing address: Qi Zhang 164 Phillips Hall, CB#3255 Department of Physics and Astronomy the University of North Carolina at Chapel Hill Chapel Hill, NC 27599-3255.
Thank you very much!
Regards, ************************************************** Qi Zhang, Ph.D 164 Phillips Hall, CB#3255 Department of Physics and Astronomy the University of North Carolina at Chapel Hill Chapel Hill, NC 27599 Tel: 919-843-2407 (Office) 919-843-0974 (EM Lab) Fax: 919-962-0480 E-mail: qizhang-at-physics.unc.edu
__________________________________ Do you Yahoo!? Yahoo! Mail - Helps protect you from nasty viruses. http://promotions.yahoo.com/new_mail
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 27 16:35:31 2004
In the book "Scanning Electron Microscopy and X-Ray Microanalysis" (by J. I. Goldstein et al., 2nd edition), you will find equations for calculating Bethe's or Kanaya Okayama range (p. 88 and 89). Here are the values for carbon as given in Table 3.2 of this book (p.89):
Hi. I am a new user of Philips scope CM300. I have used JEOL scopes. I found some difference in alignment procedures, and I have tubules with that.
1. I have troubles in understanding the relation between setting eucentric height and objective lens current in Philips scope. If I set eucentric height, it is not focus position. After I get eucentric height and I get focus position using focus nob, I can get an objective current. Objective lens current should be constant, but the obtaining objective lens current is not the same.
When I use JEOL scopes, I do not touch coarse focus nob, but change height (Z) to keep the same objective lens current. Generally, how HRTEM people control objective lens current and height in Philips scope?
2. Do HRTEM people adjust Voltage center and Coma free on CM300? Just do you do Current center and Coma free?
Thank you, Hiromi Konishi Indiana University
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 27 18:11:11 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (nbiswas-at-chem.uga.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, October 27, 2004 at 12:24:50 ---------------------------------------------------------------------------
Question: We have an Olympus BX60 microscope (upright and confocal).I need to do some temperature dependent studies of samples on glass slides for which I need a temperature range of ~20-60C. I was looking at the TS-4 and TS-4ER stages from Physitemp. I would appreciate some information on them.How good are they?If there are some other porducts please let me know.
The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------------------------
Heather -
What paper are you using in the processor. If I recall Agfa stopped making that Rapidoprint paper years ago. You could possibly use whatever chemistry for the paper you are using.
ML
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
When I called to order chemicals for our AGFA Rapidoprint Processor (AGFA Rapidoprint Activator G182B and AGFA Rapidoprint Fixer QSO4A), I was told by my supplier that AGFA is no longer manufacturing them. I called the AGFA technical support line, but they could not suggest direct replacements nor would they share the formulas for the discontinued products with me so that I can make them myself. The ingredients in the two are common photo chemicals (for the activator water, ammonium thiosulfate and sodium sulfite; for the fixer water, sodium hydroxide and sodium sulfite).
Has anyone already grappled with this problem and found suitable replacements? I could find nothing on the web and the compositions of the products are not given in the Morgan & Morgan Photo Lab Index - at least not in the vintage copy in our library.
Least you think we're in the dark ages here, we do have a film scanner. There are just some old fashioned microscopists here (mainly me!) who still prefer stacks of study prints for data analysis over stacks of CDs.
Thanks for any forthcoming solutions ;)
Heather Owen
Dr. Heather A. Owen, Director Electron Microscope Laboratory Department of Biological Sciences University of Wisconsin - Milwaukee Lapham Hall, P.O. Box 413 Milwaukee, WI 53210 USA Phone: (414)229-6816
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 28 02:31:47 2004
There should be no real difference between a JEOL TEM and FEI TEM. On a side entry machine the engineer should set the eucentric height of the stage at the optimum objective lens focus. If this is not the case I suggest you ask your engineer to check it out.
If you are saying that the focus current changes even though the eucentric height is set, this certainly should not happen and should be checked out by your engineer. A change in objective lens focus current at eucentric height could indicate a stage, lens or HT problem.
Good luck Ron
On Wed, 27 Oct 2004 17:24:31 -0500 hkonishi-at-indiana.edu wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } Hi. } I am a new user of Philips scope CM300. I have used JEOL scopes. I found some } difference in alignment procedures, and I have tubules with that. } } 1. I have troubles in understanding the relation between setting eucentric } height and objective lens current in Philips scope. If I set eucentric height, } it is not focus position. After I get eucentric height and I get focus position } using focus nob, I can get an objective current. Objective lens current should } be constant, but the obtaining objective lens current is not the same. } } When I use JEOL scopes, I do not touch coarse focus nob, but change height (Z) } to keep the same objective lens current. Generally, how HRTEM people control } objective lens current and height in Philips scope? } } 2. Do HRTEM people adjust Voltage center and Coma free on CM300? Just do you do } Current center and Coma free? } } Thank you, } Hiromi Konishi } Indiana University } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk http://www-em.materials.ox.ac.uk/ *********************************
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 28 04:29:18 2004
Is there a shelf life for probes for contact and tapping modes in AFM, How do we know that the probes used for AFM are no longer good if they are old and unused what are the signs. shashi singh CCMB Hyderabad
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From MicroscopyL-request-at-ns.microscopy.com Thu Oct 28 09:41:33 2004
To be held in conjunction with the March 24,2005 meeting of the Midwest Microscopy and Microanalysis Society
Affiliate of the Microscopy Society of America and the Microbeam Analysis Society of America
ABSTRACT DEADLINE: Abstracts must be received in electronic format by 5PM, Wednesday, December 15, 2004.
The first quarter meeting of the Midwest Microscopy and Microanalysis Society will be held on Thursday, March 24, 2005, in the Pancoe Life Sciences Building, Northwestern University, Evanston, Illinois. A student poster competition open to undergraduate and graduate students will be held in conjunction with the meeting. Posters should illustrate utilization of microscopy for either biological or materials science study. Prizes will be awarded as follows:
$300 for 1st place $200 for 2nd place $100 for 3rd place
Abstracts must be received in electronic format by 5PM on Wednesday, December 15, 2004. To be eligible for a prize, you must be first author on the poster, and you must be present at the meeting. You are encouraged to submit your entry as early as possible, as space may be limited. Further details and an explanation of judging criteria will be provided upon acceptance of your submission.
Please send the following information to the email address below, and attach your abstract as a Word document:
Name Affiliation Department Phone number Email address
Send to:
Elaine Schumacher MMMS Materials Science Director McCrone Associates, Inc. 850 Pasquinelli Drive Westmont, IL 60559-5539 Tel: 630-887-7100 Fax: 630-887-7417 Email: eschumacher-at-mccrone.com
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 28 10:03:51 2004
Dear Listers, Some time back there was a discussion about the utility of an FEG ESEM. Specifically that the resolution under ESEM conditions was not all that good and that one would do just as well with a tungsten filament ESEM. Is this the prevailing opinion of FEG ESEM users? OR are there too few instruments in service to form a consensus? Second, in terms of viewing hydrated biological samples and beam sensitive polymers and other materials, would one have better results with a cryo stage on a high vac. FEG SEM ? Respond privately or to the list.
Thanks much, Greg Erdos
Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program Scientific Director, Electron Microscopy P.O. Box 118525 217 Carr Hall University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295 fax- 352-846-0251
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 28 10:06:07 2004
Dear TEMers, We would like to try using membrane inserts to grow cultured cells and process them in situ for thin section TEM. What membrane materials have any of you used successfully?
Thanks, Greg
Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program Scientific Director, Electron Microscopy P.O. Box 118525 217 Carr Hall University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295 fax- 352-846-0251
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 28 10:30:02 2004
Agfa replaced Rapidoprint paper with Brovira-Speed paper. We use Agfa Brovira-Speed 310 Glossy, Resin Coated paper, which comes in grades 2,3, and 4 (for contrast.) I get it from Calumet (CalumetPhoto.com). We have a Rapidoprint processor and we use a developer made by Solutek Corp. (Boston.617-445-5335. Go Sox!),# RA 2201 Ready-to-use Developer, Catalog No. 536-40. Same company makes Uni-Fix Premium Fixer, Ready-to-use, Catalog No. 519-40. I found a vendor for these chemicals - J. P. Walker & Co Inc.,160 Oak Street, Glastonbury, CT, 06033, 860-633-8327. I believe they get it from a Rhode Island supplier. I realize this is not near you, but maybe they can help you.
Julie Gross Research Assistant Dept. of Neuroscience University of CT Health Center Farmington, CT. 06030 jgross-at-neuron.uchc.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 28 12:25:28 2004
Does anyone have the manuals for the LaB6 gun the DSM-962? Zeiss says they don't have any back copies. I know the SEM came with the capability to use tungsten or LaB6 and I have the right wehnalt and maybe the right anode...just don't have the manuals. Can anybody provide copies of the manuals, please?
Chuck Butterick Analytical Laboratory Supervisor Poco Graphite, Inc. 300 Old Greenwood Road Decatur, TX 76234 (940) 393-4287 (Phone) (940) 393-8383 (Fax) CButterick-at-poco.com
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 28 13:41:02 2004
-- Dr. Elaine Humphrey Director, BioImaging Facility President, Microscopy Society of Canada University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-interchange.ubc.ca website: www.emlab.ubc.ca
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 28 13:55:11 2004
Greg, I've done a LOT of samples of cell grown on polycarbonate filters. The ones from Costar work especially well (no financial interest). What's really neat is that if you cut the filters from their holders at some point during dehydration and then put them into prop. oxide after the last 100% ethanol, they roll up like a jelly roll. I usually cut them in half or thirds so that I end up with multiple blocks from one insert. I put them into flat embedding molds, let them sit in the resin for an hour of so then put them in the oven. When you trim the blocks you get a lovely spiral with many more cells/section than you would have if you'd cut up little bits of flat filter. 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
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 28 14:05:12 2004
Resolution in ESEM mode is close to resolution in high vacuum mode for "good" specimens (like Au particles). The problem is that resolution for not coated specimens is very specimen dependant. I can get good pictures of Au particles at magnification of x100k with FEG ESEM, but for hydrated hard tissue, as bone or dentin, only at x20k. I did not use magnifications above x30k for any specimens in ESEM mode, including minerals and ceramics. Hydrated soft tissues are usually covered with a thin layer of water which hides small details, so for them useful magnifications are considerably lower. As for beam sensitive specimens I think the best results could be obtained with coating or in low voltage mode (and for low voltage FEG is better).
Vladimir
} } Dear Listers, } Some time back there was a discussion about the } utility of an FEG } ESEM. Specifically that the resolution under ESEM conditions } was not all } that good and that one would do just as well with a tungsten filament } ESEM. Is this the prevailing opinion of FEG ESEM users? OR } are there too } few instruments in service to form a consensus? } Second, in terms of viewing hydrated biological } samples and beam } sensitive polymers and other materials, would one have better } results with } a cryo stage on a high vac. FEG SEM ? } Respond privately or to the list. } } Thanks much, Greg Erdos } } } Gregory W. Erdos Ph.D. } Assistant Director, Biotechnology Program } Scientific Director, Electron Microscopy } P.O. Box 118525 } 217 Carr Hall } University of Florida } Gainesville, FL 32611 } gwe-at-ufl.edu } 352-392-1295 } fax- 352-846-0251 } } }
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 28 14:33:21 2004
} Dear Listers, } Some time back there was a discussion about the utility } of an FEG ESEM. } Specifically that the resolution under ESEM conditions was } not all that good and that one would do just as well with a } tungsten filament ESEM. ...
Correct me if I am wrong, but I believe the context for that previous consensus was not for "ESEM" (i.e., 2500 Pa), but rather for "VP-SEM" (i.e, 250 Pa) ... and that FEG was somewhat useful in the the VP range of pressures(?) That is, you'll almost always find some utility in using a brighter gun .. unless high pressure unduely contaminates the FEG source.
cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 28 14:42:21 2004
} } Dear Listers, } Some time back there was a discussion about the utility of an } FEG } ESEM. Specifically that the resolution under ESEM conditions was not } all that good and that one would do just as well with a tungsten } filament ESEM. Is this the prevailing opinion of FEG ESEM users? OR } are there too few instruments in service to form a consensus? } Second, in terms of viewing hydrated biological samples and } beam } sensitive polymers and other materials, would one have better results } with a cryo stage on a high vac. FEG SEM ? } Respond privately or to the list. }
Oooooh, I hope the responses come on the list!
This is a pretty interesting topic.
Seems to me that responses should by default be on the list rather than privately, unless there are privacy/legal issues.
cheers
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 28 18:14:39 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (gtwang-at-sandia.gov) from http://www.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, October 28, 2004 at 10:33:51 ---------------------------------------------------------------------------
Email: gtwang-at-sandia.gov Name: George Wang
Organization: Sandia National Laboratories
Title-Subject: [Microscopy] [Filtered] MListserver: Opinions on FEI Quanta 3D vs JEOL 6480LV dual beam systems
Question: Hello, I am designing a system which will have as one of its modules a dual beam FIB/SEM with LV/VP/ESEM capability. Two possibilities are the FEI Quanta 3D and the JEOL JSM-6480LV w/Orsay Canion gun and gas injector system. Does anyone have any experience with or opinions about either of the two systems? Ideally, I would like to have the field emission rather than the tungsten filament column, but I doubt my budget would allow this. I'm concerned about the SEM resolution in the tungsten filament versions. Thanks, George
George T. Wang, Ph.D Senior Member of Technical Staff Chemical Processing Science Department 01126 Sandia National Laboratories P.O. Box 5800 MS 0601 Albuquerque, NM 87185 ph. 505.284.9212 fax 505.844.3211 gtwang-at-sandia.gov __________________________________________
Greg, I cannot comment on a FEG ESEM since we do not have one at present. I have worked with a W-ESEM and an SEM with cryo-stage and came to the following conclusion based on samples I have used:
1) A great strength of ESEM is the ability to do DYNAMIC experiments. In this case resolution is usually secondary. However, FEG should give better resolution than W- with APPROPRIATE samples.
2) either ESEM or low-vac can be helpful for minimizing charging when coating is not desired but this usually compromises resolution when compared to high vac imaging.
3) Hydrated samples are difficult to work with under the best of circumstances and ESEM has it's own challenges.
4) Cryo-SEM is the way to go for static hydrated imaging whenever possible. You can work with fractured samples, sublimate to remove unbound water, and work with delicate hydrated samples such as young plants with minimum problems. Resolution is normally acceptable for these types of samples with preservation far superior to the samples prepped for High Vac imaging. Charging is easily handled using coating under cryo conditions.
5) Cryo permits easy imaging of heat sensitive polymers and other delicate materials. They can be coated at low temperature if charging is a problem and resolution is not seriously compromised over standard high vacuum imaging.
6) Ultimately resolution depends much more on the sample than on the capabilities of the instrument. However, there is a greater potential to work at higher magnifications while retaining good resolution with the FEG and resulting increased beam coherence, smaller beam diameter, good signal:noise ratio, and ability to work at lower kV.
My opinion is to go with FEG whenever possible providing you can afford the increased cost of the original instrument and the service contracts. An ESEM can still benefit from FEG in high-vac mode even if there may not be a great difference in ESEM mode. Although it might not be needed for many samples now, it will be helpful for some. You never know about future needs and most of us cannot justify replacing instruments on a frequent basis.
Remember that cryo can be added to any SEM so you are not limiting that option with either ESEM or FESEM.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
On 10/28/04 10:13 AM, "Greg Erdos" {gwe-at-biotech.ufl.edu} wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------} - } } Dear Listers, } Some time back there was a discussion about the utility of an FEG } ESEM. Specifically that the resolution under ESEM conditions was not all } that good and that one would do just as well with a tungsten filament } ESEM. Is this the prevailing opinion of FEG ESEM users? OR are there too } few instruments in service to form a consensus? } Second, in terms of viewing hydrated biological samples and beam } sensitive polymers and other materials, would one have better results with } a cryo stage on a high vac. FEG SEM ? } Respond privately or to the list. } } Thanks much, Greg Erdos } } } Gregory W. Erdos Ph.D. } Assistant Director, Biotechnology Program } Scientific Director, Electron Microscopy } P.O. Box 118525 } 217 Carr Hall } University of Florida } Gainesville, FL 32611 } gwe-at-ufl.edu } 352-392-1295 } fax- 352-846-0251 } }
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 29 01:18:14 2004
The 9th European Congress on Stereology and Image Analysis will be coupled with the 7th International Conference on Stereology and Image Analysis in Materials Science STERMAT in Zakopane - Poland 2005 .
Link http://ptst.polsl.katowice.pl/9ECS/index9ECS.htm
The main topics of the meeting will be:
Applications of image analysis and stereology Recent advances in stereology New image analysis tools Young stereologists competition
Special emhasize will be devoted to:
3D data acquisition, sampling and processing Image analysis of 3D objects Medical applications of image analysis Image analysis and stereology in materials science Application of image analysis in motion control, security, automation, etc. Trainig in stereology and image analysis
The organizers keep in mind that the leading edge is currently the interface between technical and biomedical sciences. Our wish is to prepare a competent forum for exchange of ideas in the field of widely understood image analysis and stereology.
Special courses on stereology principles and basic as well as advanced skills in image analysis will accompany the meeting.
Authors of the best contributions, selected by the international advisory board, will be invited to prepare extended versions of their papers for publication in Image Analysis and Stereology or Materials Characterization.
Krzysztof Hübner Foundry Research Institute Kraków Poland
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 29 07:36:41 2004
=========================================================== The 9th European Congress on Stereology and Image Analysis will be coupled with the 7th International Conference on Stereology and Image Analysis in Materials Science STERMAT in Zakopane - Poland 2005 .
Link http://ptst.polsl.katowice.pl/9ECS/index9ECS.htm
The main topics of the meeting will be:
Applications of image analysis and stereology Recent advances in stereology New image analysis tools Young stereologists competition
Special emhasize will be devoted to:
3D data acquisition, sampling and processing Image analysis of 3D objects Medical applications of image analysis Image analysis and stereology in materials science Application of image analysis in motion control, security, automation, etc. Trainig in stereology and image analysis
The organizers keep in mind that the leading edge is currently the interface between technical and biomedical sciences. Our wish is to prepare a competent forum for exchange of ideas in the field of widely understood image analysis and stereology.
Special courses on stereology principles and basic as well as advanced skills in image analysis will accompany the meeting.
Authors of the best contributions, selected by the international advisory board, will be invited to prepare extended versions of their papers for publication in Image Analysis and Stereology or Materials Characterization.
Krzysztof H¸bner Foundry Research Institute KrakÛw Poland
{/x-charset}
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 29 14:19:02 2004
All, I have two populations of liposomes which I am examining by cryo TEM. I would like to be able to identify which liposomes are which when mixed together. I am thinking about incorporating a marker into one population of liposomes - this can be done when the liposomes are made. So far my thoughts are 1) colloidal gold 2) an electron dense stain e.g. ammonium molybdate.
Has anyone sucessfully incorporated anything into liposomes which could help with this problem?
Many thanks as always
Chris
Christopher J. Gilpin Ph.D. Assistant Professor Director, Molecular and Cellular Imaging Facility K1.246 Department of Cell Biology University of Texas Southwestern Medical Center 5323 Harry Hines Boulevard Dallas, Texas 75390-9039 +1 214 648 2827 Phone +1 214 648 6408 Fax christopher.gilpin-at-utsouthwestern.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 29 16:38:52 2004
We have a client looking at various incarnations of carbon (nanotubes, etc.) in the TEM and he is very curious about a phenomenon he is seeing. This involves dark bands traveling rapidly down lengths of carbon during viewing and is very similar to an effect I have seen commonly when looking at crystalline structures. I bet most, if not all, TEM users have noted something similar at some point, such as when looking at negatively stained specimens when part of the stain has crystallized. When the beam hits a new area, there appears to be almost a liquid flow within the crystal which stabilizes in a few seconds.
I have never given this much thought, assuming that this is just electron flow within the particles somehow affecting the beam to give this visual result. Can someone describe the physics, and possibly significance, of this?
I know I have probably described this effect very poorly, but if anyone wants to take a stab at it, let me know and I will forward a couple images to you showing the movement of these bands.
Thanks for any help. Our client seriously wants to understand this, and I don't have a clue where to start researching it (except to ask you guys!).
Happy Halloween, Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 29 18:34:10 2004
I would like to hear what solutions people have come up with for imaging cells for long times while maintaining 37C and 5-10% CO2. We have managed to get away with using CO2 Independent medium (Invitrogen) and a Bioptechs temp controller for everything up until now. We now have a situation where the cells die in this medium but are happy in RPMI in a CO2 incubator. I presume that means finding a way to maintain the CO2 on the stage. I have seen some plexiglass boxes with controllers for outrageous $$$ which can do this. Is that the best solution? Any suggestions for one to go on a Zeiss Axiovert 200? Thanks- Dave --
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 91 N. Eagleville Rd. U-3125 Storrs, CT 06269-3125 knecht-at-uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 29 19:38:07 2004
I guess it is as follows: e-beam exposure -} structure/shape change -} beam reflection shift -} change of diffraction contrast (just like moving of bend contours) -CNi
-----Original Message----- } From: Tindall, Randy D. [mailto:TindallR-at-missouri.edu] Sent: Friday, October 29, 2004 5:49 PM To: microscopy-at-microscopy.com
Hi Listers,
We have a client looking at various incarnations of carbon (nanotubes, etc.) in the TEM and he is very curious about a phenomenon he is seeing. This involves dark bands traveling rapidly down lengths of carbon during viewing and is very similar to an effect I have seen commonly when looking at crystalline structures. I bet most, if not all, TEM users have noted something similar at some point, such as when looking at negatively stained specimens when part of the stain has crystallized. When the beam hits a new area, there appears to be almost a liquid flow within the crystal which stabilizes in a few seconds.
I have never given this much thought, assuming that this is just electron flow within the particles somehow affecting the beam to give this visual result. Can someone describe the physics, and possibly significance, of this?
I know I have probably described this effect very poorly, but if anyone wants to take a stab at it, let me know and I will forward a couple images to you showing the movement of these bands.
Thanks for any help. Our client seriously wants to understand this, and I don't have a clue where to start researching it (except to ask you guys!).
Happy Halloween, Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 30 07:49:18 2004
Bioptechs and other companies (Harvard?) sell chambers for the microscope stage that are closed on the top & bottom but allow for perfusion during imaging. Changing the media this way would work.
However, we simply use a WPI heated stage or a completely enclosed forced air heat box around the micrscope for 37 degrees.
For CO2, we rent tanks of premixed 95% air / 5% CO2. We take a large petri dish and melt a hole in the side. Using thin tubing, we force the gas through two closed tubes of water to hydrate it and then into the small holein the side of the petri dish turned upside-down on top of the cells, usually in MatTek dishes, on the microscope stage. Different people in the lab have made their own personal modifications to improve the system, such as cutting a disk in a sponge and placeing the wet sponge around the dish of cells to improve hydration.
Sorry we don't have photos of this on the web yet.
-Michael
On Fri, 29 Oct 2004, David Knecht wrote:
} } } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } I would like to hear what solutions people have come up with for } imaging cells for long times while maintaining 37C and 5-10% CO2. We } have managed to get away with using CO2 Independent medium } (Invitrogen) and a Bioptechs temp controller for everything up until } now. We now have a situation where the cells die in this medium but } are happy in RPMI in a CO2 incubator. I presume that means finding a } way to maintain the CO2 on the stage. I have seen some plexiglass } boxes with controllers for outrageous $$$ which can do this. Is that } the best solution? Any suggestions for one to go on a Zeiss Axiovert } 200? Thanks- Dave } -- } } Dr. David Knecht } Department of Molecular and Cell Biology } University of Connecticut } 91 N. Eagleville Rd. U-3125 } Storrs, CT 06269-3125 } knecht-at-uconn.edu } 860-486-2200 860-486-4331 (fax) } home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html } } }
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 30 09:13:04 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (lieberw-at-mpip-mainz.mpg.de) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, October 29, 2004 at 11:16:39 ---------------------------------------------------------------------------
Organization: Max-Planck Institute for Polymer Research
Title-Subject: [Microscopy] [Filtered] MListserver: TEM data for historical microscopes covering the last 50 years
Question: Dear Listers,
i have an unusual question. I am preparing an overview on the development of TEM over the last five decades for a talk I would like to give. But i really can¥t find any data on historical TEM¥s, like resolution power, acc voltage and price. Does any of you have an idea where I can find such data?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (monica.iliescu-at-polymtl.ca) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, October 29, 2004 at 15:09:43 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] [Filtered] SC12 and SC20 Magnetic Field Cancelling Systems
Question: Hello, We must install an ESEM Quanta FEG 200, but we have a problem with the magnetic field. Right now we have a room that does not meet the specifications for magnetic field before installation (we need below 3 mG and we have 6 mG peak to peak, in the y axis, at 1 foot, 3 foot and 6 foot high). We are currently investigating different options in order to make the room meet these specifications. We heard about the SC12 and SC20 Magnetic Field Cancelling Systems (via web search) and we would like to know if somebody already utilised this kind of system. If yes (I hope) how sensitive is the system? Thank you very much! Monica
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (pakdeet2-at-uwm.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, October 29, 2004 at 16:27:26 ---------------------------------------------------------------------------
Question: I wonder if epoxy/clay nanocomposites can be cut by such regular glass knives to prepare specimens for TEM observation instead of the pricey diamond knives and what is the size and dimension of specimen should it be,thank you
Question: I wonder if epoxy/clay nanocomposites can be cut by such regular glass knives to prepare specimens for TEM observation instead of the pricey diamond knives and what is the size and dimension of specimen should it be, thank you ============================================== The clay particles will just "laugh" at the glass knife when it sees it coming. You can't do it that way.
Now for a word about "pricey". If you ever have the opportunity to go through a diamond knife production facility, you would probably be wondering why they don't cost more.
But if you do want to save money, SPI Supplies as well as some of the other suppliers of diamond knives offer "materials science" diamond knives which are on average, about 35% lower in cost than a so-called "life science" diamond knife. Different vendors define their "materials science" knives differently, but in the case of SPI, the only difference is that the knife edge has a few slight striations that would cause it to flunk as a life science knife but for a materials science sample they are just fine. The reason is that on the first pass with the knife on your sample, you are putting in striations far more profound than the few that are present when the knife is new.
Disclaimer: SPI Supplies was the first firm in the industry (1976) to offer commercially a materials science diamond knife and we would like to see more people using them!
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 30 16:49:27 2004
Monica: I'm using as SC12 Field Canceling system right now on my JEOL 6600FXV. It's canceling my fields to below 0.15 mG. I don't have the specs handy right now but it should be able to handle your fields. My system was installed by Vibration Engineering Consultants (http://www.vibeng.com/), who sell and service Spicer units in North America. Now I'm just waiting for my active vibration canceling system to get rid of the 15 Hz shake in my building.
by way of MicroscopyListserver wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-648-8743 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 30 17:12:18 2004
} Email: pakdeet2-at-uwm.edu } Name: Choakchai Pakdeethai } } Organization: University of Wisconsin-Milwaukee } Education: Graduate College } Location: Milwaukee , WI USA } } Question: I wonder if epoxy/clay nanocomposites can be cut by such } regular glass knives to prepare specimens for TEM observation } instead of the pricey diamond knives and what is the size and } dimension of specimen should it be,thank you } } --------------------------------------------------------------------------- Choakchai:
I agree with Chuck Garber's comments about the need for a diamond, but the rest of your question leaves me wondering about both your sample and your level of knowledge of microtomy. Is your sample already embedded? If so, did you follow a protocol specific for clay? It's easy to change its ultrastructure with the wrong prep method. If you're a novice at microtomy, please don't try to teach yourself; get help from an experienced microtomist in Milwaukee, or attend Boeckeler-RMC's materials microtomy workshop in Tucson next March.
Caroline -- Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 30 20:21:56 2004
Can anyone point me toward a good reference on insect (primarily caterpillar) anatomy and morphology? I am particularly interested in gut morphology.
Thanks, Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
From MicroscopyL-request-at-ns.microscopy.com Sun Oct 31 16:00:15 2004
Here is the November 2004 Microscopy Today table of contents. This issue ships with the Call for Papers and poster for the M&M-2005 meeting.
I will close the subscription list for this issue on Friday, 5 November 2004. Microscopists in North America and MSA members anywhere can subscribe for free. Anyone else may subscribe for US$35 per year (to PARTIALLY cover postage). All subscriptions at http://www.microscopy-today.com Thank you.
Finding the First Fires with Microscopes Stephen W. Carmichael, Mayo Clinic
Combining Imaging and Spectroscopy: Solving Problems with Near Infrared Chemical Imaging E. N. Lewis, E. Lee, and L. H. Kidder, Spectral Dimensions, Inc.
Automated Functions in Electron Microscopy Bill Tivol, California Institute of Technology
Variable Magnification Electron Holography for 2‑D Mapping of Semiconductor Devices Y.Y. Wang, M. Gribelyuk, A. Domenicucci, J. Bruley, J. Gaudiello, and M. Kawasaki,* IBM Microelectronic Division, *JEOL USA
A FIB Micro-Sampling Technique for Three-Dimensional Characterization of a Site-Specific Defect T. Yaguchi1, Y. Kuroda1, M. Konno1, T. Kamino1, T. Ohnishi2, T. Hashimoto2, K. Umemura2, K. Asayama3; 1Hitachi Science Systems, 2Hitachi High-Technologies Corp., 3 Renesas Technology Corp., Japan
Color Matching to Ink Jet Printers from a Computer Screen, Part 1 Jerry Sedgewick, University of Minnesota
FIB Lift-Out and Milling of Cylindrical Specimens for Electron Tomography (or Atom Probe Field Ion Microscopy) Lucille A. Giannuzzi, FEI Company
A New Improved Silicon Multi-Cathode Detector (SMCD) for Microanalysis and X-Ray Mapping Applications S. Barkan*, V. D. Saveliev*, J. S. Iwanczyk*, L. Feng*, C. R. Tull*, B. E. Patt*, D. E. Newbury**, J. A. Small**, and N. J. Zaluzec;*** *Radiant Detector Technologies, **National Institute of Standards and Technology, ***Electron Microscopy Center, Argonne National Lab
Broadband CARS Microscopy Marcus T. Cicerone and Tak W. Kee, National Institute of Standards and Technology
Microwave Processing of Drosophila Tissues for Electron Microscopy JoAnn Buchanan, Stanford University School of Medicine
Mechanical Polishing Methods for Metal Samples for EBSD S. Roberts*, D. Flatoff*, B. True;** *South Bay Technology, Inc., **EDAX / TSL
High Pressure Freezing User Group Meeting at M & M 2004
The Microscopy Society Of America Project MICRO Caroline Schooley, Project MICRO Coordinator
Industry News
MSA 2005 Undergraduate Research Scholarships
NetNotes
Ron Anderson, Editor Microscopy Today
From MicroscopyL-request-at-ns.microscopy.com Sun Oct 31 23:10:01 2004
I would like to embed soil in place in the field to compare the disposition of organic matter, roots, microbes, etc in the soil among various cropping systems with as little disruption to the soil as possible so it can be studied on both macro and micro levels.
In western Oklahoma I can find the soil when it is extremely dry of free water nearly every summer. Of course that is a long way from free of water but I understand there are some embedding resins that are very tolerant of water.
Does anyone have a protocol that will work for this. I don't expect to get very much penetration into the face of the soil. A very few mm would be great.
Thanks
Gordon
Gordon Couger (retried) Biosystems & Agricultural Engineering Oklahoma State University 624 Cheyenne Ave gcc (at) couger.com Stillwater, OK 74075 405 624 2855 cell 405 269 3588 After 3:00 PM Central Time.
From MicroscopyL-request-at-ns.microscopy.com Fri Nov 5 08:05:15 2004
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jose.maria.manero-at-upc.es) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, November 5, 2004 at 07:34:24 ---------------------------------------------------------------------------
Email: jose.maria.manero-at-upc.es Name: Jose M Manero
Organization: UPC
Title-Subject: [Microscopy] [Filtered] How to prepare samples to see barcteria?
Question: Dear Colleagues
I have samples of tooth decay. My goal would be to have a look on the existing bacteria in the caries through an electron microscopy. Samples were observed by me by means of an ESEM (without any kind of preparation), as well as I have observed them by a SEM (just gold coated samples) and I have never been able to see them. Has anyone experience in how to prepare samples to see the bacteria? Do I have to use conventional methods: deshydratation, fixation, critical points, conductive coatingsÖ?
Than you in advance for your co-operation!
Dr. Jose M Manero Electron Microscopy Laboratory Departament of Materials Science UPC. Av Diagonal 647. Barcelona. Spain
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (steven.obenour-at-unison.ae.ge.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, November 5, 2004 at 07:19:48 ---------------------------------------------------------------------------
Email: steven.obenour-at-unison.ae.ge.com Name: Steven Obenour