Sasaki, S., J.K. Steven & N. Bodick (1983). Serial reconstruction of microtubula array within dendrite of the cat retinal ganglion cell: the cytoskeleton of a vertebrate dendrite. Brain Res. 259, 193-206.
Stevens, J.K. & J.T. Trogadi (1984). Computer-assisted reconstruction from serial electron micrographs: a tool fo the systemati stud of neuronal form and function. Adv. Cell. Neurobiol. 5, 341-369.
Yours, -Dick Gordon
At 3:09 P -0500 9/30/03, b wa of MicroscopyListserve wrote: } ------------------------------------------------------------------------------ } The Microscop ListServe -- Sponsor: The Microscop Societ of America
-- .................................................................................................. Dr. Richard Gordon, Depts. of Radiology/Electrical & Compute Engineering, U.Manitoba, Room GA216, Health Science Centre, 820 Sherbrook Street, Winnipeg, MB R3A 1R9 Canada Adjunct Scientist: TRLabs, Mentor: IEEE/EMBS Student Chapter Scientist, Manitoba Institute of Child Health. Editorial Board: Journal of Mechanic in Medicine and Biology; Home page: http://www.umanitoba.ca/faculties/medicine/radiology/stafflist/rgordon.html Lab: (204) 789-3828, Fax: (204) 787-2080, E-mail: GordonR-at-ms.UManitoba.ca Reprint Hunter/Professo Finder: http://www.umanitoba.ca/faculties/medicine/radiology/search/universities.html
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 1 08:19:54 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rsimmons-at-gsu.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, October 1, 2003 at 07:59:29 ---------------------------------------------------------------------------
Question: You might have a look at the LR resins (LR White / LR Gold) from London Resin Co. I believe that they will work well with your fluorescence application and you can thin section them as well if you need to.
Joel, It may not be your resin that is fluorescing, but the glutaraldehyde you fixed with. For light microscopy, we quench that fluorescence with one of the following used after the post-fix rinse: 0.1 M glycine 50 mM ammonium chloride or sodium borohydride (I hate using that one, so I don't have a recipe at my finger tips) 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 Wed Oct 1 09:13:38 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jwilkinson-at-seton.org) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Wednesday, October 1, 2003 at 08:59:11 ---------------------------------------------------------------------------
Question: I am currently working in a EM lab in a hospital. I need a procedure on how to process ciliary biopsies. After we receive the brushing from surgery. Can you help me?
Hi there, I am looking for spare screens for my JEOL 2010F, they are JEOL part numbers 607071 (small screen - focussing screen) and 607072 (large screen - viewing screen) respectively. If anyone has any spare I would be happy to buy them. JEOL have none in stock and have a 45 day lead time on them and so I was wondering if anyone is sat on a spare pair. Reply off line please.
-- John Mansfield PhD MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 USA Phone: (734) 936-3352 FAX (734) 763-2282 Cell. Phone: (734) 834-3913 (Leaving a phone message at 936-3352 is preferable to 834-3913) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42° 16' 48" Long. 83° 43' 48"
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 1 09:31:01 2003
Hi there, I'm trying again to see if anyone is interested in starting a JEOL 2010F user group. We have some questions on how other users configure things on their microscope and also have some tips that we would like to share with other users. I tried doing this before but we were tied up in the installation of the scope and I never pursued it. Let me know if you are interested offline and I will see if I can get a mailing list together at least. Thanks in advance.
-- John Mansfield PhD MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 USA Phone: (734) 936-3352 FAX (734) 763-2282 Cell. Phone: (734) 834-3913 (Leaving a phone message at 936-3352 is preferable to 834-3913) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42° 16' 48" Long. 83° 43' 48"
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 1 09:34:36 2003
} Subject: [Microscopy] IASTED Modelling and Simulation Newsletter - Sept. 2003 } } IASTED International Newsletter on Modelling and Simulation } September 30, 2003 } } RECENTLY ANNOUNCED MODELLING AND SIMULATION CONFERENCES } 1. The 15th IASTED International Conference on Modelling and Simulation - MS 2004 } March 1-3, 2004, Marina Del Rey, CA, USA } } Important Deadlines: } Submissions Due: Oct. 15, 2003 } Notification of Acceptance: Nov. 15, 2003 } Registration Deadline: Dec. 15, 2003 } } To submit a paper, tutorial or special session or for registration information visit our website at http://www.iasted.com/conferences/2004/marina/ms.htm } } 2. The 4th IASTED International Conference on Modelling, Simulation, and Optimization - MSO 2004 } August 16-18, 2004, Kauai, Hawaii, USA } Important Deadlines: } Submissions Due: Feb. 15, 2004 } Notification of Acceptance: Apr. 15, 2004 } Registration Deadline: June 1, 2004 } To submit a paper, tutorial or special session or for registration information visit our website at http://www.iasted.com/conferences/2004/hawaii/mso.htm } } UPCOMING CONFERENCE DEADLINES } The submission deadline for the 23rd IASTED International Conference on Modelling, Identification and Control - MIC 2004 is Oct. 15, 2003. MIC 2004 is being held Feb. 23-25, 2004 in Grindelwald, Switzerland. Delegates without papers are also welcome to register until Dec. 15, 2003. http://www.iasted.com/conferences/2004/switzerland/mic.htm } } } FUTURE CONFERENCES } Mark your calendars! The IASTED International Conference on Environmental Modelling and Simulation will be held Nov. 22-24, 2004 in St. Thomas, Virgin Islands, USA. } } } MODELLING AND SIMULATION JOURNALS FROM ACTA PRESS } The International Journal of Modelling and Simulation - First published in 1981, this journal covers all aspects of modelling, simulation, languages, software, hardware, methodology, numerical and graphical methods, virtual reality, statistical techniques, tutorials, surveys, and applications. It al so includes book reviews, conference notices, call for papers, and new publications. } Editor-in-Chief: Prof. A. Houshyar } Frequency: 4 issues per year } 2003 Rate: US$256.00 } Postage & Handling: US$25.00 } ISSN: 0228-6203 (205) } http://www.actapress.com/journals/journals.htm#Modelling } } } CONFERENCE PROCEEDINGS AVAILABLE } Past conference proceedings in the area of modelling and simulation are available for purchase from ACTA Press - http://www.actapress.com/proceedings/proceedings.htm } } For more information, to be removed from our mailing list, or to join one of the following mail groups: Power, Telecommunication, Bio-Medicine, Signal and Image Processing, Artificial Intelligence, Business, Software Engineering, Education, Databases and Knowledge Engineering, Internet and Applications, Parallel and Distributed Computing, please contact: } IASTED } #80, 4500 - 16th Avenue N.W. } Calgary, Alberta } Canada T3B 0M6 } Tel: 403-288-1195 } Fax: 403-247-6851 } E-mail: calgary-at-iasted.com } Web site: http://www.iasted.org }
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 1 09:36:34 2003
The Fall Meeting of the Michigan Microscopy and Microanalysis Society will be held October 17, 2003 at the Kellogg Hotel and Conference Center in Lansing, Michigan.
Oral and poster sessions will be held in this one day conference. Abstract submission deadline was September 19 but late submissions are welcome. You are encouraged to attend or submit a paper. For further information or to submit an abstract, contact:
Ginam Kim President Michigan Microscopy and Microanalysis Society (MMM) Tel. 989-496-5077 or e-mail to g.kim-at-dowcorning.com
Kevin Battjes MMM Secretary Impact Analytical Voice 989-832-5555, ext 556 Michigan Molecular Institute Fax 989-832-5560 1910 W. St Andrews Road e-mail: battjes-at-mmi.org Midland MI 48640 battjes-at-impactanalytical.com
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 1 09:37:42 2003
Sasaki, S., J.K. Stevens & N. Bodick (1983). Serial reconstruction of microtubular arrays within dendrites of the cat retinal ganglion cell: the cytoskeleton of a vertebrate dendrite. Brain Res. 259, 193-206.
Stevens, J.K. & J.T. Trogadis (1984). Computer-assisted reconstruction from serial electron micrographs: a tool for the systematic study of neuronal form and function. Adv. Cell. Neurobiol. 5, 341-369.
Yours, -Dick Gordon
At 3:09 PM -0500 9/30/03, by way of MicroscopyListserver wrote: } ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- .................................................................................................. Dr. Richard Gordon, Depts. of Radiology/Electrical & Computer Engineering, U.Manitoba, Room GA216, Health Sciences Centre, 820 Sherbrook Street, Winnipeg, MB R3A 1R9 Canada Adjunct Scientist: TRLabs, Mentor: IEEE/EMBS Student Chapter Scientist, Manitoba Institute of Child Health. Editorial Board: Journal of Mechanics in Medicine and Biology; Home page: http://www.umanitoba.ca/faculties/medicine/radiology/stafflist/rgordon.html Lab: (204) 789-3828, Fax: (204) 787-2080, E-mail: GordonR-at-ms.UManitoba.ca Reprint Hunter/Professor Finder: http://www.umanitoba.ca/faculties/medicine/radiology/search/universities.html
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 1 10:07:33 2003
I didn't see, or prematurely deleted, the original post but if glut is being used, that will cause problems with fluorescence. We use paraformaldehyde for all of our tissue fixation/fluorescent applications with no trouble.
Cheers
----------------------------------------------------------------------- Christopher S. Zurenko Research Assistant II Kresge Hearing Research Institute, Otopathology The University of Michigan Medical School MSRB 3, Room 9303 1150 W. Medical Center Dr. Ann Arbor, MI 48109-0648 Lab Phone: 734.763.9680 Fax: 734.615.8111 czurenko-at-umich.edu http://www.khri.med.umich.edu/research/raphael_lab/index.htm
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 1 10:34:54 2003
The way that I process nasal brushing is to pellet the cells in microcentrifuge tubes, (many if necessary), overlay with 3% glutaraldehyde for a hour or so, gently unstick the pellet and process as a tissue block. You will continually lose some cells, but the majority will stay together. The trouble with this method is that ciliated cells will be far and few between, thus many grids will be necessary to obtain an adequate number of cells.
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 {jwilkinson-at-seton.org} 10/01/03 10:12AM } } }
------------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jwilkinson-at-seton.org) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Wednesday, October 1, 2003 at 08:59:11 ---------------------------------------------------------------------------
Question: I am currently working in a EM lab in a hospital. I need a procedure on how to process ciliary biopsies. After we receive the brushing from surgery. Can you help me?
Thanks for all the replies about the autofluorescence. I did quench glutaraldehyde with sodium borohydride. Any other suggestions? Thanks
jmancuso wrote: Hi all, I am looking for a non-autoflourescent resin. I am labeling specific cells that are not synchronized. The cells grow on ACLAR, and they are fixed with glut, and OsO4, embedded on a piece of ACLAR with the cells facing down into the resin. This creates a thin layer of resin with cells close or directly on the surface. I'm using DAPI to stain the nucleus / chromosomes to identify cells of interest. I can see cells (although it is had to focus) using a light microscope, but the resin seems to be auto-fluorescent. Currently i have only tried Epon-812. Has anybody tried this before? Thanks in advance.
Joel Mancuso Department of Molecular and Cell Biology 345 LSA University of California Berkeley, CA 94720-3200
p - 510.643.8277 email - jmancuso-at-uclink.berkeley.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 1 11:18:54 2003
Dear Joel, We've been doing something similar with Spurr's epoxy for quite a while with paraformaldehyde fixed tissue, avoiding the use of glut. Both plastics seem to have as little autofluorescence as any others I've tried. I've gotten around the autofluorescence problem in two ways. One, collect a z-series through your sample by widefield fluorescence and use deconvolution. A nearest neighbors can be sufficient for this purpose, as well as the most time efficient algorithm. the other approach is confocal, provided you can get access to a system with UV or a 405 nm laser diode. Even excitation at 351 nm didn't present much autofluorescence by confocal.
The glut. will add problems with autofluorescence that are worst when exciting in the UV or blue wavelengths. perhaps you could use a nuclear stain excitable in the visible range.
We have a paper accepted by Brain Research Protocols related to this, but don't yet have a publication date.
Regards, Glen
On Tuesday, September 30, 2003, at 10:43 AM, jmancuso 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, } I am looking for a non-autoflourescent resin. I am labeling specific } cells } that are not synchronized. The cells grow on ACLAR, and they are fixed } with } glut, and OsO4, embedded on a piece of ACLAR with the cells facing } down into } the resin. This creates a thin layer of resin with cells close or } directly on } the surface. I'm using DAPI to stain the nucleus / chromosomes to } identify } cells of interest. } I can see cells (although it is had to focus) using a light } microscope, but } the resin seems to be auto-fluorescent. Currently i have only tried } Epon-812. } Has anybody tried this before? } Thanks in advance. } } Joel Mancuso } Department of Molecular and Cell Biology } 345 LSA } University of California } Berkeley, CA 94720-3200 } } p - 510.643.8277 } email - jmancuso-at-uclink.berkeley.edu } } } Glen MacDonald Core for Communication Research Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 USA (206) 616-4156 glenmac-at-u.washington.edu
************************************************************************ ****** The box said "Requires Windows 95 or better", so I bought a Macintosh. ************************************************************************ ******
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 1 11:42:25 2003
Joyce, we also use the centrifugation method, then mix the pellet with a drop of warm 2% agar, let harden and cut into pieces for osmication and embedding. You may find of value my "cilia principles" when scoping:
IMMOTILE CILIA SYNDROME (ICS)
Key Info for diagnosis (Dec 2001)
Based on my reading of Mierau et al 1992 (MIE92) and other pertinent articles which point out the controversies and problems, I have reached the following conclusions re making a diagnosis one way or the other on a cilia specimen where ICS may be suspected:
1. For a diagnosis of ICS, the issues of "Universality and Permanence" must be met i.e. several sites and a repeat bx are required since there are cases where dynein arms have been absent one time but present another.
2. For a diagnosis of "complete dynein arm absence" (ICS according to some authors), negative findings i.e. absence of all arms on 50-100 cilia examined in perfect cross section allow only for a "consistent with" type of statement since some pts may have normal axonemes on repeat bx (MIE92)
3. For a "rule out" diagnosis i.e. reporting the occurrence of normal cilia, several well preserved cilia with some inner and outer arms must be seen. Inner arms are usually blurred and less distinct than outer arms. (DIC00). Rarely are all arms seen in a given cross section since successive arms within (i.e. through the depth of) the thin section must be superimposed to be seen (MIE92). Under the best circumstances, only about 20% of inner arms are seen. Dickersin (Dic00) p771 contends that the normal average number of outer/inner arms per axoneme is two or more. But, keep in mind that Ghadially (GHA88) reports that in ICS pts, occasional cilia can show some arms, usually short. Therefore, I conclude that it is best to find some arms in all or most of a dozen or more cilia examined in perfect cross section.
REFERENCES
MIE92: Mierau GW, et al 1992. The role of electron microscopy in evaluating ciliary dysfunction: report of a workshop. Ultrastructural Pathology 16:245-254.
DIC00: Dickersin GR, 2000. Diagnostic Electron Microscopy 2nd ed, Springer
GHA88: Ghadially F, 1988 Ultrastructural Pathology of the Cell and Matrix. erd ed, p1200, Butterworths
Tom Christensen Director EM Lab Pathology Boston U Med Center
} --------------------------------------------------------------------------- } } Email: jwilkinson-at-seton.org } Name: Joyce Wilkinson } } Organization: Brackenridge Hospital } } Education: Undergraduate College } } Location: Austin, Texas USA } } Question: I am currently working in a EM lab in a hospital. } I need a procedure on how to process ciliary biopsies. After we receive the brushing from surgery. Can you help me? } } } ---------------------------------------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 1 12:02:48 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (william.treasurer-at-exeloncorp.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, October 1, 2003 at 10:50:37 ---------------------------------------------------------------------------
Email: william.treasurer-at-exeloncorp.com Name: William Treasurer
Organization: Exelon PowerLabs
Title-Subject: [Microscopy] MListserver: HNU EDS System
Question: We have a HNU Model 5000, Ser # AX2001 EDS System. Is there any companies that still provide service on these systems?
We are located in Willington, IL. My phone number is 815-458-7654.
I gather from your note that you are embedding with the ACLAR. Is it possible that this is the source of your autofluorescence?
Many years ago, when growing cells on polystyrene culture dishes, I experimented with fixing the cells, and then dissolving the plastic with ethylene dichloride. The wisps of cells were repelleted and embedded. It might work for you.
} } } ---------------------------------------------------------------------- } -------- 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 again, } } Thanks for all the replies about the autofluorescence. I did quench } glutaraldehyde with sodium borohydride. Any other suggestions? Thanks } } } jmancuso wrote: } Hi all, } I am looking for a non-autoflourescent resin. I am labeling specific } cells that are not synchronized. The cells grow on ACLAR, and they are } fixed with glut, and OsO4, embedded on a piece of ACLAR with the cells } facing down into the resin. This creates a thin layer of resin with } cells close or directly on the surface. I'm using DAPI to stain the } nucleus / chromosomes to identify cells of interest. I can see cells } (although it is had to focus) using a light microscope, but the resin } seems to be auto-fluorescent. Currently i have only tried Epon-812. } Has anybody tried this before? } Thanks in advance. } } Joel Mancuso } Department of Molecular and Cell Biology } 345 LSA } University of California } Berkeley, CA 94720-3200 } } p - 510.643.8277 } email - jmancuso-at-uclink.berkeley.edu } }
Joel B. Sheffield, Ph.D. Biology Department, Temple University 1900 North 12th Street Philadelphia, PA 19122 jbs-at-temple.edu (215) 204 8839, fax (215) 204 0486 http://astro.temple.edu/~jbs
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 1 15:20:02 2003
.. dusting off a very old Email message out of the archives....
} } --------------------------------------------- } } Date: Fri, 1 Oct 1993 15:18:05 -0500 (CDT) } } From: ZALUZEC-at-ANLEMC.MSD.ANL.GOV (Nestor J. Zaluzec (708)-252-5075, -4964) } } Message-Id: {931001151805.20201c44-at-ANLEMC.MSD.ANL.GOV} } } Subject: [Microscopy] Open for Business } } To: Microscopy-at-anlemc.msd.anl.gov } } X-Vmsmail-To: MICROSCOPY } } } } } } The Microscopy Listserver/Mailreflector is } } now running you may begin posting messages } } } } Cheers- Nestor } } } } ---------------------------------------------
Well after a decade of (very nearly) continuous operation, we've survived/outlived a number of servers (the original VAX-785, several Sun's, and now a PC running Linux), a few domain names and address changes, and had several different sponsors (MSA has obviously been the longest running), and I'll have to admit alot of headaches.
Many of you will recall that we delivered our 100 Millionth Email message about two years ago, and the searchable archives occupy ~ 120 Mbytes of on-line storage. Lately have been averaging ~ 18 Gbits/month of Microscopy Email (and that is not counting the junk mail!).
There was no junk/spam mail back in '93, today the ratio is around over 10:1, fortunately junk is for the most part blocked, albeit at the expense of occassionally rejecting some real messages in error.
A number of the original subscribers from back in Oct of 93 are still members of this list. I glad this means I'm not the only die hard in group. I unfortunately did not keep a complete copy of that initial list of subscribers, at least not on any media that I can still read and process here at home. I'll have to go back into my old files at ANL and see if I can resurrect a very old tape drive for that job. It would be interesting to see how many of the original members are still subscribed. I certainly have grown to recognize alot of the Email addresses on the list, and I'll have to admit having met alot of you over the years.
I do, however, remember that Arthur Day from ANSTO (ard-at-ansto.gov.au) was the first subscriber, while john chandler (chandler-at-lamar.ColoState.EDU) replied to the first question on the list. Want to guess who posted that first question ? ;-)
I'll have to admit that back in '93 I would have never predicted that I would still be running this server 10 years later.
In any event, as I hoist a virtual pint in the air ... onto the next decade..
Cheers....
Nestor Your (suddenly feeling tired) Friendly Neighborhood SysOp
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 1 15:25:35 2003
I have done astrocytes stained with GFAP+fluorescine and embedded in EMBed 812, no problems. Fixation was with 4% paraformaldehyde though, not glut.
Geoff
Joel Mancuso wrote:
} Hello again, } } Thanks for all the replies about the autofluorescence. I did quench } glutaraldehyde with sodium borohydride. Any other suggestions? Thanks } } } jmancuso wrote: } Hi all, } I am looking for a non-autoflourescent resin. I am labeling specific cells } that are not synchronized. The cells grow on ACLAR, and they are fixed with } glut, and OsO4, embedded on a piece of ACLAR with the cells facing down into } the resin. This creates a thin layer of resin with cells close or directly on } the surface. I'm using DAPI to stain the nucleus / chromosomes to identify } cells of interest. } I can see cells (although it is had to focus) using a light microscope, but } the resin seems to be auto-fluorescent. Currently i have only tried Epon-812. } Has anybody tried this before? } Thanks in advance. } } Joel Mancuso } Department of Molecular and Cell Biology } 345 LSA } University of California } Berkeley, CA 94720-3200 } } p - 510.643.8277 } email - jmancuso-at-uclink.berkeley.edu } } } }
-- -- ********************************************** 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 Wed Oct 1 15:39:18 2003
On Friday, September 26, 2003, at 08:32 AM, Ephram Shizgal wrote:
} any suggestions of TEM spec prep for free flowing polymers ? } Dear Ephram, I don't know what the viscosity of your polymers might be, but cryo-TEM may be possible if you can blot enough of the polymer to leave a sufficiently thin layer. Alternatively, you could cool the polymer until it is solid, then thin it or microtome it as one would with other solid polymers. Yours, Bill Tivol 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 1 15:50:05 2003
Nestor, Looking back over the last 10 years with all the long hours, headaches and God knows what else I'd like to suggest that you hoist not a virtual pint but a real one. If you haven't already done so I'm sure you're planning it!
Cheers, Paul
Paul J. Gerroir Microscopy Materials Characterization Xerox Research Centre of Canada 2660 Speakman Drive Mississauga, Ontario L5K 2L1
-----Original Message----- } From: Nestor J. Zaluzec [mailto:zaluzec-at-microscopy.com] Sent: Wednesday, October 01, 2003 4:19 PM To: microscopy-at-ns.microscopy.com
Nestor: I think I can speak for most all of us subscribers in saying that's it's been a decade of extremely GREAT service by our Friendly Neighborhood SysOp.
"Nestor J. Zaluzec" 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 } ------------------------------------------------------------------------------- } } .. dusting off a very old Email message out of the archives.... } } } } } --------------------------------------------- } } } Date: Fri, 1 Oct 1993 15:18:05 -0500 (CDT) } } } From: ZALUZEC-at-ANLEMC.MSD.ANL.GOV (Nestor J. Zaluzec (708)-252-5075, -4964) } } } Message-Id: {931001151805.20201c44-at-ANLEMC.MSD.ANL.GOV} } } } Subject: [Microscopy] Open for Business } } } To: Microscopy-at-anlemc.msd.anl.gov } } } X-Vmsmail-To: MICROSCOPY } } } } } } } } } The Microscopy Listserver/Mailreflector is } } } now running you may begin posting messages } } } } } } Cheers- Nestor } } } } } } --------------------------------------------- } } Well after a decade of (very nearly) continuous operation, we've } survived/outlived a number of servers (the original VAX-785, } several Sun's, and now a PC running Linux), a few domain } names and address changes, and had several different sponsors } (MSA has obviously been the longest running), } and I'll have to admit alot of headaches. } } Many of you will recall that we delivered our 100 Millionth Email } message about two years ago, and the searchable archives occupy } ~ 120 Mbytes of on-line storage. Lately have been averaging } ~ 18 Gbits/month of Microscopy Email (and that is not counting the } junk mail!). } } There was no junk/spam mail back in '93, today the ratio is around } over 10:1, fortunately junk is for the most part blocked, albeit at } the expense of occassionally rejecting some real messages in error. } } A number of the original subscribers from back in Oct of 93 } are still members of this list. I glad this means I'm not the only } die hard in group. I unfortunately did not keep a complete copy of that } initial list of subscribers, at least not on any media that I can still read } and process here at home. I'll have to go back into my old files at ANL } and see if I can resurrect a very old tape drive for that job. } It would be interesting to see how many of the original members } are still subscribed. I certainly have grown to recognize alot of the Email } addresses on the list, and I'll have to admit having met alot of you over } the years. } } I do, however, remember that Arthur Day from ANSTO (ard-at-ansto.gov.au) } was the first subscriber, while john chandler (chandler-at-lamar.ColoState.EDU) } replied to the first question on the list. Want to guess who posted that } first question ? ;-) } } I'll have to admit that back in '93 I would have never predicted that I } would still be running this server 10 years later. } } In any event, as I hoist a virtual pint in the air ... onto the next decade.. } } Cheers.... } } Nestor } Your (suddenly feeling tired) Friendly Neighborhood SysOp } }
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 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 Wed Oct 1 16:28:23 2003
So come Savannah, let's each of us buy Nestor a pint. Phil
} .. dusting off a very old Email message out of the archives.... } } } } } --------------------------------------------- } } } Date: Fri, 1 Oct 1993 15:18:05 -0500 (CDT) } } } From: ZALUZEC-at-ANLEMC.MSD.ANL.GOV (Nestor J. Zaluzec } } } (708)-252-5075, -4964) } } } Message-Id: {931001151805.20201c44-at-ANLEMC.MSD.ANL.GOV} } } } Subject: [Microscopy] Open for Business } } } To: Microscopy-at-anlemc.msd.anl.gov } } } X-Vmsmail-To: MICROSCOPY } } } } } } } } } The Microscopy Listserver/Mailreflector is } } } now running you may begin posting messages } } } } } } Cheers- Nestor } } } } } } --------------------------------------------- } } Well after a decade of (very nearly) continuous operation, we've } survived/outlived a number of servers (the original VAX-785, } several Sun's, and now a PC running Linux), a few domain } names and address changes, and had several different sponsors } (MSA has obviously been the longest running), } and I'll have to admit alot of headaches. } } Many of you will recall that we delivered our 100 Millionth Email } message about two years ago, and the searchable archives occupy } ~ 120 Mbytes of on-line storage. Lately have been averaging } ~ 18 Gbits/month of Microscopy Email (and that is not counting the } junk mail!). } } There was no junk/spam mail back in '93, today the ratio is around } over 10:1, fortunately junk is for the most part blocked, albeit at } the expense of occassionally rejecting some real messages in error. } } A number of the original subscribers from back in Oct of 93 } are still members of this list. I glad this means I'm not the only } die hard in group. I unfortunately did not keep a complete copy of that } initial list of subscribers, at least not on any media that I can still read } and process here at home. I'll have to go back into my old files at ANL } and see if I can resurrect a very old tape drive for that job. } It would be interesting to see how many of the original members } are still subscribed. I certainly have grown to recognize alot of the Email } addresses on the list, and I'll have to admit having met alot of you over } the years. } } I do, however, remember that Arthur Day from ANSTO (ard-at-ansto.gov.au) } was the first subscriber, while john chandler (chandler-at-lamar.ColoState.EDU) } replied to the first question on the list. Want to guess who posted that } first question ? ;-) } } I'll have to admit that back in '93 I would have never predicted that I } would still be running this server 10 years later. } } In any event, as I hoist a virtual pint in the air ... onto the } next decade.. } } Cheers.... } } Nestor } Your (suddenly feeling tired) Friendly Neighborhood SysOp } } }
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 1 16:36:51 2003
Are you a classroom volunteer? Or might you try being one, if you knew a bit about how to really stimulate youngsters' interest? Project MICRO's "Microscopic Explorations" is one approach, and another very effective one is "Private Eye", which introduces observation of the microworld with cheap jeweler's loupes. If you live in the Los Angeles area, you can attend a "Private Eye" workshop at the California State Teachers Association meeting at the Long Beach Convention Center - Friday October 10, 12-1, room 301, or Saturday October 11, 10:30-11, room Seaside A; you don't need to register for the meeting.
The "Private Eye" workshops are excellent, and there will be future opportunities in other cities. If you're interested, contact me, or "Private Eye" directly at dmelody-at-theprivateeye.com. -- 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/ Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 1 16:36:19 2003
An editor of one of the National Science Teacher's Association magazines has just contacted me with a request for a short article; I know that several of you listers enjoy writing short essays, so how about it? She has an unreasonably short deadline - Oct. 15. Please remember that the readers will be elementary school teachers! Contact the editor directly {valynda_m-at-nsta.org} . ----------------------------------- At 11:55 AM -0400 10/1/03, Valynda Mayes wrote: } I am writing from the National Science Teachers Association in search of a } scientist to write a short piece for our journal for elementary teachers, } Science and Children. "Science 101" explains a topic in a question and } answer format and is written by a specialist working in the chosen field. } } In our next issue, we are publishing an article about an upper elementary } classroom using a digital microscope to observe the reproductive parts of } plants. The question we'd like to address in "Science 101" is "How do } microscopes work?" We are looking for a brief explanation appropriate for } the elementary science classroom, with a few interesting tidbits thrown in, } such as examples of ways microscopes are used in science. In research on } this topic, I came across the Project MICRO website and thought you might be } able to assist us in finding someone interested in writing this piece. } } I usually try to locate scientists with an interest in children and } education who have an affinity for this sort of thing. The piece is short } (about 400 words) and we offer an honorarium. It is our hope that "Science } 101" will introduce professions to our readers and their students in } addition to providing interesting background content. We include a short bio } and photograph of the scientist at the end of each article. If you can be of } help, please be in touch. } } Thank you for your time, } } Valynda Mayes } Assistant Editor } Science and Children
-- 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/ Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 1 17:02:03 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (lesleykelley11-at-hotmail.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, October 1, 2003 at 15:34:52 ---------------------------------------------------------------------------
Email: lesleykelley11-at-hotmail.com Name: Lesley
Organization: ExxonMobil Chemical
Title-Subject: [Microscopy] MListserver:
Question: We are looking at purchasing a new FESEM next year. Any input on current models would be greatly appeciated.
We are currently looking at SEMs from LEO, JEOL and Hitachi.
Please respond to lesley.a.kelley-at-exxonmobil.com
Good job Nestor. Many thanks from all of us. If you will be at UCLA I'll provide beer or vodka (your choice). Sergey.
At 02:27 PM 10/1/2003, 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 33-089 Los Angeles, CA 90095
Joel Sheffield wrote: ====================================================== I gather from your note that you are embedding with the ACLAR. Is it possible that this is the source of your autofluorescence?
Many years ago, when growing cells on polystyrene culture dishes, I experimented with fixing the cells, and then dissolving the plastic with ethylene dichloride. The wisps of cells were repelleted and embedded. It might work for you. ================================================================ We have been selling ACLAR® cut sheets (as have been others as well) for some number of years and a substantial number of those using it find one of its main advantage to be its virtual freedom from any autofluorescence. This is described on our URL http://www.2spi.com/catalog/photo/aclar-film.shtml
Obviously there could in theory be some low level autofluorescence from the ACLAR film, but it is apparently too low to be measured (at least by most users of the film).
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 2 03:51:02 2003
I haven't used the Brunel version but it looks like a generic Cambridge rocking microtome only grey instead of black. If it is built to the Cambridge standard then it will probably last forever. The only thing that I've ever had to replace on our 15 or so Cambridge units is the occasional spring but that seems to be after about 15 or more years of student use. Our models also have a cord to attach the end of the arm to the specimen feed ratchet mechanism and this can wear out after many years but we found that we could get a replacement locally.
These microtomes are very simple and will happily cut between 4 and 10+ um wax (and I assume plastic) sections. Specimen orientation is rudimentary you rotate or move the specimen backwards or forwards to line up by releasing the front clamp. I found that a disposable blade system was more convenient than the solid steel blade but that might depend on your facilities for sharpening microtome knives.
If this microtome is for student use, research or irregular use then it will be ideal but if you need to cut hundreds of sections in long 'ribbons' then a rotary microtome would be better but much more expensive.
One further issue is that because the knife is exposed at the front it would be advisable to get into a strict routine of removing it from the microtome when not in use or fashioning a cover to prevent accidents.
Please again note that my comments are about Cambridge rocking microtomes (many of ours must be 30+ years old by now). But these are easily repaired and maintained because of their simple mechanisms.
Malcolm
Malcolm Haswell e.m. unit School of Health, Natural and Social Sciences University of Sunderland Tyne & Wear SR1 3SD UK e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- } From: "max_gra-at-libero.it" {max_gra-at-libero.it}
We are in the process of sorting out the safety of the lab. This was a question I could not answer. A good website I used was: http//www.healthcouncil.nl
Thanks
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 2 05:03:51 2003
I am a new user in EM and I am having a difficult time getting my Epoxy sections to stick to the copper grids. I have tried Acetic acid washes, sonicators, HCl wash etc. I have not tried BSA, I am waiting for the order to come in. Does anyone have any more suggestions?
Thank you in Advance. Sue
-- Sue Tyler Biologist Cooperative Oxford Laboratory Center for Coastal Environmental Health& Biomolecular Research at Charleston ( CCHEBR) USDOC/NOAA/NOS/NCCOS 904 S. Morris St. Oxford, Maryland 21654-9724 410-226-5193 Fax: 410- 226-5925 Sue.Tyler-at-noaa.gov
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 2 08:31:41 2003
I realize that bone is bone, but I'm trying to differentiate between new bone and old bone in paraffin sections of bone with implants. I'm looking for a method of determining which areas are "old bone" and which are "new" (the new being those areas where bone has started to grow from the insertion of the implant). I have tried basic H&E, 0.5% aq. toluidine blue, and a Van Gieson's method. I thought somebody might have a suggestion, but if not, it was worth a try!
Thanks so much,
Elke Pravda Biostructure Core Facility Forsyth Institute Boston, MA 02115
epravda-at-forsyth.org
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 2 11:23:57 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (slevinso-at-ic.sunysb.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, October 2, 2003 at 10:14:53 ---------------------------------------------------------------------------
Email: slevinso-at-ic.sunysb.edu Name: Sarah Zimov
Organization: Stony Brook University
Title-Subject: [Microscopy] MListserver: Ultrastructural localization of Ca
Question: Hello, I was just wondering if anyone was familiar with the oxalate precipitation method or the osmium/potassium-bichromate method for visualizing calcium- at the EM level. Particularly is it important to fix in Glutaraldehyde? I am trying to visualize calcium in an intracellular store in retinal/nervous tissue. If anyone has any suggestions on the best method to use and/or a good working protocol it would be greatly appreciated!!!!!
Sarah Zimov Neurobiology and Behavior Stony Brook University Stony Brook, NY 11794
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (opto-at-klughammer.de) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, October 2, 2003 at 10:07:15 ---------------------------------------------------------------------------
Central States Microscopy and Microanalysis Society Fall Meeting: October 24, 2003
The fall meeting of the Central States Microscopy and Microanalysis Society will be held at the Donald Danforth Plant Science Center in St. Louis, on Friday, October 24, 2003. Our keynote speaker will be Dr. James Pawley of the University of Wisconsin Department of Zoology, speaking on "The 3-methods of 3D microscopy: which one to choose". Dr. Pawley's presentation will be sponsored by the Microscopy Society of America Tour Speaker Program and by CSMMS.
We are also soliciting presentations on all aspects of microscopy and microanalysis, including electron microscopy, confocal and fluorescence microscopy, x-ray microanalysis, materials, geological, and biological microscopy applications, immunocytochemistry, and related topics. Students, faculty, and staff are all welcome to make short presentations of about 10-30 minutes in length, and we are planning a competition with cash prizes for best student presentation.
Details on the meeting schedule, as well as directions to the Danforth Plant Science Center and links to information about the Center, can be found at: http://www.danforthcenter.org/imf/csmms/index.htm. As the schedule firms up, more information will be added.
Many thanks to Howard Berg of the Danforth Center and to Heather Ford, previously of the Center, for their efforts at organizing the meeting at this great venue. A tour of the Center will be offered at the meeting. Dr. Berg may be contacted at: RHBerg-at-danforthcenter.org.
Randy Tindall, President Central States Microscopy and Microanalysis Society
Electron Microscopy Core 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.biotech.missouri.edu/emc/
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 2 13:51:39 2003
On Thursday, October 2, 2003, at 02:55 AM, Coetzee, Mr S. H Physics Science wrote:
} We are in the process of sorting out the safety of the lab. This was } a question I could not answer. A good website I used was: } http//www.healthcouncil.nl } Dear Mr. Coetzee, OsO4 is very toxic as a vapor, so heating it would, presumably, increase the concentration of vapor, hence increase toxicity. If, however, the fire is in a reducing condition, i.e., there is so much flammable material that oxygen content is very low, then the OsO4 could be reduced to a less toxic compound. I would not base safety considerations on speculation about the state of the fire, but I'd treat heated OsO4 as a very serious safety hazard. Yours, Bill Tivol 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 Thu Oct 2 14:17:28 2003
I posed a similar question recently. I too have had section adhesion issues.
I didn't have a EMS grid coating pen (which I have since purchased) so I tried dissolving different types of tape and adhesive in chloroform (I didn't have any benzene or ethylene dichloride).
My adhesives are untested as for actual section adhesion (maybe next week), but I did view naked grids with a dried drop of adhesive solution under a brightfield/light scope to see how well the grid was coated. I felt that 3-4 Avery Spot-O-Glue tabs transferred to something else (I used a SEM stub) then dissolved in 10 mL chloroform produced the most clean, uniform coating of adhesive.
The pen is a $33 item and a few people suggest it but I think I'm still going to try my home brew to see if it works.
Regards,
----------------------------------------------------------------------- Christopher S. Zurenko Research Assistant II Kresge Hearing Research Institute, Otopathology The University of Michigan Medical School MSRB 3, Room 9303 1150 W. Medical Center Dr. Ann Arbor, MI 48109-0648 Lab Phone: 734.763.9680 Fax: 734.615.8111 czurenko-at-umich.edu http://www.khri.med.umich.edu/research/raphael_lab/index.htm
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 2 15:12:15 2003
Sue, I have found that after picking up sections if I put the grids in the 50-60 degree oven for several hours to overnight, the sections always stick. It isn't necessary to clean them first either. (I use the thin bar copper grids from EMS) Good luck, Mary Gail Engle
Mary Gail Engle Sr. Research Laboratory Manager Electron Microscopy & Imaging Facility Health Sciences Research Bldg. 001 University of Kentucky Lexington, KY 40536-0305
phone 859-323-6108 fax 859-257-9700
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 2 15:12:59 2003
I don't use this type of equipment, so I can't comment on it, but Gatan sent me some literature a while back with SEM hot stages that I still happen to have around. 3 models: H1001, H1002, H1005 Max temp.: 500, 750, 1500 (deg. C), respectively Hope this helps ------------------------------------------------ Kevin Frischmann, Laboratory Manager Microscopy & Imaging Facility American Museum of Natural History Central Park West at 79th Street New York, NY 10024-5192 USA
At 11:21 AM 10/2/03 -0500, opto-at-klughammer.de wrote:
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From MicroscopyL-request-at-ns.microscopy.com Thu Oct 2 15:20:54 2003
Krzysztof Herman wrote: ========================================================== Does somebody know direct contact to manufacturer of good dry boxes for instruments like cameras, lenses, prec, mechanics....
But this is probably from fifth hand.... ====================================================== The Secador line of desiccating cabinets represents an economical yet very highly effective means of storage of delicate optical parts be they parts for LM or even EM instrumentation or cameras. They can be found on URL http://www.2spi.com/catalog/supp/desiccator-cabinets-secador.html
The larger models come with an innovative electrically driven desiccant regeneration system so that one never needs to worry about desiccant replacement.
One of these cabinets should work quite nicely in your intended application depending on how much capacity you require. And how much automation you need. The 220v units are CE certified.
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 2 15:50:48 2003
The Fall meeting of the New England Society for Microsocpy (NESM) will be held on Tuesday, October 14th at the headquarters of Boston Scientific Corporation in Natick, MA.
The meeting will feature dinner and two technical presentations in materials science. Dr. Francesco Stellacci will speak on "The Use of the Scanned Probe Microscope as a Lithographic Tool" and Steve Stokowski will speak on "Black Granite from Hell". Preregistration is strongly encouraged.
For complete program, registration deadline, etc. re: this meeting, please go to NESM's website: http://prism.mit.edu:8083/Newsletters/September2003Newsletter.pdf
Peggy Sherwood Corresponding Secretary, NESM
Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of 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 2 16:13:56 2003
You are invited to participate to a Workshop on Advanced Techniques in Scanning Electron Microscopy & Microanalysis for materials Characterization. That workshop will be held from May 17 to 21, 2004 at McGill University, Montreal, Canada.
The lecturers are:
Raynald Gauvin, McGill University, Canada
Pierre Hovington, Hydro-Québec Research Center, Canada David C. Joy, University of Tennessee and Oak Ridge National Laboratory, USA Marin Lagacé, Hydro-Québec Research Center, Canada Eric Lifshin, State University of New York at Albany, USA
For more information:
http://www.minmet.mcgill.ca/ebeamworkshop
Since there is a limit in the number of participants, please register early if you are interested.
Very best regards
Raynald GAUVIN ************************************************************ Professor Raynald Gauvin Department Mining, Metals and Materials Engineering McGill University M.H. Wong Building 3610 University Street Montreal, H3A 2B2 Canada Tel: (514) 398-8951 FAX: (514) 398-4492 http://www.minmet.mcgill.ca EMAIL:Raynald.Gauvin-at-McGill.ca ************************************************************
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 2 22:27:16 2003
Hello all I am interested in following the fate of our biodegradable microparticles over time, when incubated in aqueous solutions such as plasma. We have a laser diffraction particle sizer, but I would like to be able to look at structural changes in wet mounts by microscopy, ideally monitoring particle diameter with good enough statistics to correlate the LM result with the laser diffraction result. I am interested not just in average size, but the distribution of sizes. Thus ideally I'd like to automate the measurements of particle diameter. Our particles are ~ 15 microns diameter on average, encapsulating aqueous solutions, and the particles (being denser than the suspending medium, which must be aqueous and isotonic) sink to the bottom of a wet mount. Just like latex beads they act as lenses in brightfield, so becke lines (bright & dark rings) are visible and vary with the plane of focus, which makes it hard to define the actual particle's edge. Using a low NA objective & closing the iris to maximize depth of field doesn't seem to eliminate the becke lines. There can be a range of sizes of particles present simultaneously in a field, so (at least when using higher NA optics) when I am focused on the equator of a 30 micron particle I am far away from the equator of a 10 micron particle, and vice versa. I may be able to increase the refractive index of the suspending medium to minimize the becke lines, so the bigger problem in my mind is the variation in height (above the surface of the slide) of the particles present.
Because of these considerations, I wonder if anyone has devised a way to accurately size a _mixture_ of, say, 10 um & 30 um calibrated latex beads in water, settled on the slide, by microscopy. It seems tricky to do; I can't imagine doing it with a single image (containing many particles) from a single focal plane. I can imagine 3 ways: 1) autofocus on each bead, find the plane of the equator & image that; 2) take a z-series of a field, for each particle find the slice with greatest diameter and take measurements for that particle only from that slice; or 3) full-on 3D reconstruction & characterization. I would need to accumulate data from many hundreds of particles, but with many particles per field it may not require that many fields. It doesn't seem that MetaMorph, Image-Pro Plus, or ImageJ is up to this level of automation or analysis but I could certainly be mistaken; I'm not familiar with any of these. I would appreciate suggestions for how to approach this, using either conventional or confocal microscopy.
Thanks! Richard
Richard Thrift SkyePharma, Inc 10450 Science Center Drive San Diego, CA, 92121 USA (858) 625-2424
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 3 05:35:06 2003
A simple way to do this is to give tetracycline to the animals in the period which you think the new bone is being formed. The tetracycline will be deposed in the new bone and will be fluorescente in green (FITC filter) under a fluorescent microscopy and you will be able to differentiate the new bone from the old bone.
I wish you good luck. Sincerely Prof. Dr. Francisco Javier Hernandez Blazquez Universidade de São Paulo Fac. de Medicina Veterinária e Zootecnia Departamento de Cirurgia - Setor de Anatomia Av. Prof. Dr. Orlando Marques de Paiva, 87 05508-000 - São Paulo (SP) - Brasil Tel..55 (11) 3091 1374 Fax 55 (11) 3091 7805 email: fjhblazq-at-usp.br
----- Original Message ----- } From: "Pravda, Elke" {epravda-at-forsyth.org} To: {Microscopy-at-msa.microscopy.com} Sent: Thursday, October 02, 2003 10:30 AM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (karla.schuster-at-delta.com) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Friday, October 3, 2003 at 13:14:27 ---------------------------------------------------------------------------
Email: karla.schuster-at-delta.com Name: karla schuster
Education: Graduate College
Location: Marietta, GA, USA
Question: I'm thinking of a microscope as a birthday present for my daughter (8 yrs). Can you recommend 2-3 good microscopes?
I've read several sites on that tell you what to look for, but I'm not familiar with good manufacturers.
Since it is pretty clear that that you will not be measuring the same particle every time, is there some reason why you cannot take a "representative" sampling of the particles remove them from the liquid and then image them dry?
Once they are dry then you have a variety of approaches (possibly even low voltage SEM) that might improve things.
Nestor Your Friendly Neighborhood SysOp
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 4 00:27:21 2003
It isn't a must do now, fast track, immediate deal, but I need to begin assembling a list (particularly in the NE, USA) of qualified confocal service companies, individuals, technicians, and/or contract people capable of entering into a service agreement with a major university on a Zeiss confocal META LSM-510 for a long term contract.
If anyone has any suggestions, referrals, or underpaid or out of work Zeiss confocal tekkies please let me know
Please forward them or refer them to me.
Gratefully,
Bruce Grosso www.AssetRecovery.Net, Inc. 67 County Rd. 274 Iuka, MS 38852 662-423-1757 Ph. 662-423-1299 Fx bgrosso-at-AssetRecovery.Net Equipment Recovery Specialists
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 4 06:30:41 2003
I received an answer at my questions about the Brunel microtome. Unlikely I deleted that mail before I read it completely. I remember that he was telling me that he had bought the microtome but he had not jet used it because he have had to finish the wax inclusion. I apologize but could who answer me to re-mail his answer. Thank you, Best Regards,
Massimo T.
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 4 18:05:35 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (florea-lupu-at-omrf.ouhsc.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Saturday, October 4, 2003 at 09:07:29 ---------------------------------------------------------------------------
Organization: Oklahoma Medical Research Foundation
Title-Subject: [Microscopy] Techni G2 12TWEEN vs Hitachi H7600
Question: We are interested to buy a new electron microscope and we have short listed two instruments: Techni G2 12TWEEN and Hitachi H7600. I would appreciate opinions about the reliability and performances of these microscopes from colleagues who used both instruments (not sales representatives!) A swift response would be very helpful,
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kolb-at-uni-mainz.de) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, October 5, 2003 at 13:18:52 ---------------------------------------------------------------------------
Dear Sarah, Oxalate precipitation method is quite good for visualizing calcium. You have to fix the tissue no matter what you use- glut or paraformaldehyde. You can only avoid osmium. The protocol is-- Infuse or perfuse your tissue with oxalate sol and fix it with GLu or paraformaldehyde. Dehydrate and embedd in resin. cut section and scan grids without staining with UA and L Citrate. with regards Shashi CCMB, Hyderabad INDIA
--- by way of MicroscopyListserver {slevinso-at-ic.sunysb.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 } ------------------------------------------------------------------------------- } } Below is the result of your feedback form } (NJZFM-ultra-55). It was submitted by } (slevinso-at-ic.sunysb.edu) from } http://microscopy.com/MicroscopyListserver/MLFormMail.html } on Thursday, October 2, 2003 at 10:14:53 } --------------------------------------------------------------------------- } } Email: slevinso-at-ic.sunysb.edu } Name: Sarah Zimov } } Organization: Stony Brook University } } Title-Subject: [Microscopy] MListserver: } Ultrastructural localization of Ca } } Question: Hello, } I was just wondering if anyone was familiar with the } oxalate precipitation method or the } osmium/potassium-bichromate method for visualizing } calcium- at the EM level. Particularly is it } important to fix in Glutaraldehyde? I am trying to } visualize calcium in an intracellular store in } retinal/nervous tissue. If anyone has any } suggestions on the best method to use and/or a good } working protocol it would be greatly } appreciated!!!!! } } Sarah Zimov } Neurobiology and Behavior } Stony Brook University } Stony Brook, NY 11794 } } --------------------------------------------------------------------------- }
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From MicroscopyL-request-at-ns.microscopy.com Mon Oct 6 08:32:23 2003
Hello everyone, I've recently inherited a LKB glass knife maker. I've used these beast before, but not the Type 7801B. If anyone has a set of instructions for this unit, would you please contact me and fax me a copy?
Thanks!!!!
Frank Karl Degussa Corp. frank.karl-at-degussa.com Fax 330-668-3846
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 6 09:42:17 2003
At 01:53 PM 10/5/2003 -0500, you wrote: } Email: kolb-at-uni-mainz.de } Name: Ute Kolb } } Organization: University Mainz Germany } } Title-Subject: [Microscopy] MListserver: HAADF image quantification } } Question: Dear Electronmicroscopists, } } I wonder if anyone has tried to quantify HAADF images and if there is } anything published on this subject.
There certainly is. The earliest papers on ADF-STEM by Albert Crewe (Science 168, 1338 (1970)) contain attempts at quantification. More recently, M. M. J. Treacy and Rice have quantified the contrast of atoms and small particles on a support (J. Microscopy 156, 211 (1989)). And in a series of several papers, the Silcox group at Cornell has studied quantification of high-resolution HAADF images of zone axis crystals. Here's a more or less random collection of papers:
Hillyard, S. E. & Silcox, J. (1995). Detector geometry, thermal diffuse scattering and strain effects in ADF STEM imaging. Ultramicroscopy 58, 6-17.
Kirkland, E. J., Loane, R. F. & Silcox, J. (1987). Simulation of annular dark field STEM images using a modified multislice method. Ultramicroscopy 23, 77-96.
Loane, R. F., Kirkland, E. J. & Silcox, J. (1988). Visibility of single heavy atoms on thin crystalline silicon in simulated annular dark field. Acta. Cryst. A44, 912-927.
Loane, R. F., Xu, P. & Silcox, J. (1991). Thermal vibrations in convergent-beam electron diffraction. Acta. Cryst. A47, 267-278.
Muller, D. A., Edwards, B., Kirkland, E. J. & Silcox, J. (2001). Simulation of thermal diffuse scattering including a detailed phonon dispersion curve. Ultramicroscopy 86, 371-380.
My own recent efforts in that direction (Voyles, Muller, & Kirkland, Ultramicroscopy 96, 251 (2003) and to appear in the December issue of Microscopy & Microanalysis) are a direct extension of the Silcox results.
Best wishes, Paul Voyles
Paul Voyles Assistant Professor Materials Science and Engineering Department University of Wisconsin - Madison 1509 University Ave. Madison, WI 53706-1595 Voice: (608) 265-6740 Fax: (608) 262-8353 voyles-at-engr.wisc.edu www.engr.wisc.edu/mse/faculty/voyles_paul.html
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 6 09:44:16 2003
The oxalate precipitation method does show calcium deposits but one has no way of assessing their physiological relevance. Oxalate doesn't precipitate the calcium until it is inside the cell and it doesn't get inside the cell until it is fixed by the aldehydes (which takes many seconds to many minutes depending on the tissue, fixative and cellular location). This allows massive re-distribution of calcium. The only valid way to look at calcium distribution in fixed tissues is in quick-frozen (high pressure freezing or metal mirror slam freezing) followed by freeze-drying. Freeze-substitution might be an acceptable alternative depending on the solvent and processing conditions. Even with this rigorous approach, redistribution can occur if you cut your thin sections on a water medium; sectioning on glycerol can help (see Dudek & Boyne 1986 Am J Anat 175:217). It is not a trivial problem to localize physiological ions in fixed cells.
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Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
I came across a problem concerning some specific sample preparation for TEM. The samples include (1) Precipitated Silica, (2) Silica sol and (3) Silica dispersed in polystyrene. As I am more familiar with only biological sample preparation, I would really appreciated your help in this regards.
Thanking you in advance.
Jos Nongkynrih
Sophisticated Analytical Instrumentation Facility
North Eastern Hills University
Bijni complex
Shillong 793003
Meghalaya, India
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From MicroscopyL-request-at-ns.microscopy.com Mon Oct 6 13:47:08 2003
Hi, I've just been informed by JOEL that the JOEL field service engineer in our area may be tied up with a major new installation for the next 2 months. I have a JOEL6300 SEM that I'm hoping to have installed in a newly renovated lab beginning in a about two or three weeks. Are there any independent SEM service engineers out there that are qualified to install JOEL SEM's, particularly the 6300/6400 series?
TIA Mike -- ******************************************************************** Michael M. Cheatham 312 Heroy Geology Laboratory Phone (315)-443-1261 Syracuse University Fax (315)-443-3363 Syracuse, NY 13244-1070
owner of PLASMACHEM-L: http://listserv.syr.edu/archives/plasmachem-l.html owner of XRF-L: http://listserv.syr.edu/archives/xrf-l.html owner of TIMS-L: http://listserv.syr.edu/archives/tims-l.html owner of SIRIS-L: http://listserv.syr.edu/archives/siris-l.html ********************************************************************
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 6 15:45:15 2003
On Thu, 2 Oct 2003, Mary Gail Engle wrote: } I have found that after picking up sections if I put the grids in the 50-60 } degree oven for several hours to overnight, the sections always stick. It } isn't necessary to clean them first either. (I use the thin bar copper } grids from EMS)
I use the same grids and find that 10 minutes in my 60-65 degree oven is long enough to do the trick to help in adhesion.
Karen Bovard Creighton University Medical Center Department of Pathology Omaha, Nebraska
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 6 18:50:38 2003
Another factor that affects section adhesion is the method in which the sections are picked up. If the sections are picked up by immersing the grid and coming from underneath, or picked up in a loop and placed on top of the grid, they adhere much better than if the grid is pushed down on them from above.
Ralph Common Electron Microscopist Michigan State University Division of Human Pathology A608 East Fee Hall East Lansing, MI 48824 517-355-7558; fax 517-432-1053 ralph.common-at-ht.msu.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 6 20:06:21 2003
PLEASE CONTACT ME IMMEDIATELY IF YOU HAVE EXPERIENCE SERVICING THE
ZESS LSM-510 META CONFOCAL MICROSCOPE.
Bruce Grosso www.AssetRecovery.Net, Inc. 67 County Rd. 274 Iuka, MS 38852 662-423-1757 Ph. 662-423-1299 Fx bgrosso-at-AssetRecovery.Net Equipment Recovery Specialists
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 6 21:26:30 2003
Our clients, a college of medicine and a Zeiss LSM service and repair facility have asked us to screen potential candidates to work in the Columbia, Maryland area as a service and repair technician for the Zeiss LSM-510 META Microscope being placed in a college of medicine in the greater New York City area.
We are looking for, on behalf of our client, a fully trained and qualified service and repair technician familiar with all aspects of the maintenance, repair, software, installation and upkeep of the LSM-510.
The screening is being done by AssetRecovery.Net, Inc. due to the fact that we are providing a; what is a, now fully functional, scope to our buyer and as a part of that agreement we need to locate with the intention of recruiting an LSM-510 level service and repair technician to the service company being asked to deliver a service contract to the School for this scope.
All qualified applicants will be immediately referred to the service contractor for final evaluation.
Duties include operation, maintenance, installation and upgrading of light microscope instrumentation; utilization of associated digital imaging system; specimen preparation; and methods development.
This individual will manage a centralized LSM (currently, a new Zeiss LSM 510) located in the New York area, and will provide confocal services, consultation, training, and access for user operation to researchers and other Zeiss qualified LSM service technicians.
Please submit resume to:
Bruce Grosso www.AssetRecovery.Net, Inc. 67 County Rd. 274 Iuka, MS 38852 662-423-1757 Ph. 662-423-1299 Fx bgrosso-at-AssetRecovery.Net Equipment Recovery Specialists
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 6 21:27:34 2003
Our clients, a college of medicine and a Zeiss LSM service and repair facility have asked us to screen potential candidates to work in the Columbia, Maryland area as a service and repair technician for the Zeiss LSM-510 META Microscope being placed in a college of medicine in the greater New York City area.
We are looking for, on behalf of our client, a fully trained and qualified service and repair technician familiar with all aspects of the maintenance, repair, software, installation and upkeep of the LSM-510.
The screening is being done by AssetRecovery.Net, Inc. due to the fact that we are providing a; what is a, now fully functional, scope to our buyer and as a part of that agreement we need to locate with the intention of recruiting an LSM-510 level service and repair technician to the service company being asked to deliver a service contract to the School for this scope.
All qualified applicants will be immediately referred to the service contractor for final evaluation.
Duties include operation, maintenance, installation and upgrading of light microscope instrumentation; utilization of associated digital imaging system; specimen preparation; and methods development.
This individual will manage a centralized LSM (currently, a new Zeiss LSM 510) located in the New York area, and will provide confocal services, consultation, training, and access for user operation to researchers and other Zeiss qualified LSM service technicians.
Please submit resume to:
Bruce Grosso www.AssetRecovery.Net, Inc. 67 County Rd. 274 Iuka, MS 38852 662-423-1757 Ph. 662-423-1299 Fx bgrosso-at-AssetRecovery.Net Equipment Recovery Specialists
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 7 08:05:45 2003
Dear TEM friends, Thank you so much for all of the grid adhesion suggestions I can now write a book! I will give them all a try and let you know what works for me. Thank you, Sue -- Sue Tyler Biologist Cooperative Oxford Laboratory Center for Coastal Environmental Health& Biomolecular Research at Charleston ( CCHEBR) USDOC/NOAA/NOS/NCCOS 904 S. Morris St. Oxford, Maryland 21654-9724 410-226-5193 Fax: 410- 226-5925 Sue.Tyler-at-noaa.gov
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 7 08:59:55 2003
I have the following position available in St. Louis MO. It is a one year contract position. Position is with a major Bio-Tech firm.
The contract researcher will be responsible for the in-house transmission electron microscopy experiments on nanophase materials/heterogeneous catalysts, including acquiring proper TEM images, diffraction data, and spectra as well as the interpretation of these experimental results in relation to catalyst structure, performance, and synthesis parameters/methodologies. The selected candidate will interact with multifunctional groups of scientists working on various materials/catalysts projects. This is a post-doctoral position.
Thank you,
Dennis Madden 314-432-5833 800-351-5369 Q & C International, Inc. 11330 Olive Boulevard Suite 218 St. Louis, MO 63141 dmadden-at-qcintl.com http://www.qcintl.com
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 7 10:05:54 2003
The University of Wisconsin Materials Science Center is looking for a staff member to see to the care and feeding of our TEMs and TEM users. UW is a great place to work, and Madison is a great place to live (if you don't mind cold), so I encourage qualified people to apply. The official ad with all the details is given below.
If you would like to apply, DO NOT reply to this email. Instead, contact Rick Noll (noll-at-engr.wisc.edu, or the information given below).
Sincerely, Paul Voyles --------------
The Materials Science Center at the University of Wisconsin invites applications for an electron microscopist to oversee its transmission electron microscopy facility. The facility includes high resolution and energy filtered imaging, STEM, EDS, EELS, and sample preparation. It supports a broad range of research by students, staff, and faculty with interests in materials characterization. The candidate's responsibilities will include user training and assistance, instrument maintenance, and instrument development.
Candidates must have a strong background in the theory and practice of TEM microcharacterization of materials with a minimum of 2 years post-graduate experience. The candidate should also have a strong background in vacuum technology, electronics, and computers. Strong interpersonal and communications skills are essential. Candidates should have an MS (PhD preferred) in a physical science or engineering discipline. Candidates with a BS in physical science or engineering with professional experience and knowledge in the required field equivalent to an MS will also be considered. Candidates are encouraged to view additional information at http://www.ohr.wisc.edu/pvl/pv_045842.html.
Interested candidates should send a resume and cover letter referring to Position Vacancy Listing #45842 to Richard Noll, 1509 University Ave., Madison, WI 53706, or email to noll-at-engr.wisc.edu.
To ensure full consideration, all information should be received by November 30, 2003. Note that unless confidentiality is requested in writing, information regarding the names of applicants must be released upon request. Finalists cannot be guaranteed confidentiality.
UW-Madison is an equal opportunity/ affirmative action employer. We promote excellence through diversity and encourage all qualified individuals to apply.
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 7 10:10:30 2003
Hello all, A friend recently acquired a dinosaur of a cryostat and is interested in trying to find a manual or someone with knowledge of the instrument. Any help would be appreciated. All relevant info is below and responses may be addressed directly to him, or I will forward if posted on the list. Thanks much, Jay
} International Equipment Company (IEC) } Needham Heights, Mass. } Model CTD - International - Harris - Cryostat } } Model CTD serial No. 69835M-5 } H.P. 1/5 Volts 115 AC 60 cycles } 4 amps } Refrigerant R-12 10 oz. } } International minot custom microtome } No. 69835M - 5 } } Please see what information you can find on this equipment and if you } know someone that we could hire to come in here and help us set it up. } } Thanks, } } Steven Hahn } Materials Engineer } Kelch Corporation } A Bemis Manufacturing Company } N85 W12545 Westbrook Crossing } P.O. Box 9025 } Menomonee Falls, WI 53052-9025 } Phone: 262.257.7029 } Fax: 262.257.7128 } email: stevehahn-at-kelch.com
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 7 10:35:17 2003
Does anyone have an Olympus BX51 with a phototube? We don't around here, and the Olympus information is unclear as to: mount, C-mount or bayonet? at the top, and if there is a projection/transfer lens within the tube, and if so (there must be), where exactly and what power? 1? 0.32? etc. Thanks.
Phil
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 7 11:03:17 2003
We need a double tilt holder for our JEM-100CXII. If you want to sell your one or give it away, or if you know the website on which I can look for the used TEM stuffs, please let me know. I remembered that someone mentioned the website for second-hands TEM Stuffs, but I forgot to keep it. Many thanks.
Regards, Qi
*************************************************** 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 Fax: 919-962-0480 E-mail: qizhang-at-physics.unc.edu qizhang-at-email.unc.edu
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From MicroscopyL-request-at-ns.microscopy.com Tue Oct 7 12:46:08 2003
Howdy. I recently began a new job in which I am the scientist responsible for our TA Instruments micro-TA 2990 AFM. I have never used an AFM before, let alone this particular one (my background is in STEM/TEM), so I don't know how or where to begin. I have been instructed by the person previously in this position (at a new company now) not to "play" with the AFM until I have been trained, which won't be until first quarter 2004. We will need to use this equipment in the next three months, so it is imperitive that I get up to speed on this technique.
Does someone have experience with this particular AFM that might be able to help me get started? Alternatively, I could arrange to visit a relatively local lab (I am in Union Beach, NJ, generally in the NYC metro area) that has a micro-TA for at least rudimentary instruction on aligning the system and such. As a third option, I could buy lunch for a user able to come here to get me started.
Anyone who can assist, I look forward to hearing from you. Thanks.
Kirk
Brian (Kirk) Kirkmeyer, Ph.D. Materials Characterization Lab International Flavors and Fragrances 1515 State Highway 36 Union Beach, NJ 07735-3542 732-335-2426 732-335-2350 FAX brian.kirkmeyer-at-iff.com
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 7 16:49:23 2003
The BX51 with a trinoc head has a female dovetail port for photos. Depending on what type of camera you want to use, that will determine the camera port adapter that you would need.
If you use a SLR bayonet (Nikon, etc.) camera, then there is no photo eyepiece (phototube). If you use the old or new PM-series film camera systems, they mount directly and require a photo eyepiece. These are from 2.5, 3.3, 4 and 5X. 3.3 is the normal mag photo eyepiece. If your camera is C-mount and is a typical 1/2" CCD, then get the Olympus U-TVO.5XC adapter. It allows setting parfocality. Other types of adapters for different cameras may also require a photo eyepiece.
What type of camera are you going to use?
gary g.
At 08:33 AM 10/7/2003, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Tue Oct 7 18:14:12 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (evdargelis-at-esi-il.com) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, October 7, 2003 at 16:48:50 ---------------------------------------------------------------------------
Email: evdargelis-at-esi-il.com Name: Ed Dargelis
Organization: Engingeering Systems, Inc., 3851 Exchange Ave., Aurora, IL 60504
Title-Subject: [Microscopy] MListserver: SEM Impression Materials
Question: Hello Listers, I'm interested to know if anyone has experience using dental impression materials (vinyl polysiloxanes) or mold/impression brands such as Struers or GE for replicating surfaces for SEM examination. Thanks kindly for your time and info.
Ed Dargelis evdargelis-at-esi-il.com
Engineering Systems, Inc. 3851 Exchange Ave. Aurora, IL 60504
At last I have finished to build my hand microtome. It works quite well, although the slices are not so thin as I expected. I think I have to recover the clearance between the cylinder, moved by a micrometer screw, and its seat. For cutting I´m using an USA razor blade. But all that concerns the improvement of the device. For testing it, I tried to cut a little wax block without speciemen included. The trouble is that the thin wax film rolls up when the blade goes forward in the cutting direction. The microtome worked at room temperature, about 20°C. The lukewarm water doesn't stretch the wax roll, and if I try to open it with needles the wax crumbles. So I wonder if someone could give me some suggestion to overcame this problem. Thank you. Best Regards,
Massimo T.
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 8 10:06:01 2003
I have used the polymer molding materials extensively for making replicas of surfaces for microscopic examination. The Struers material is very easy to use and makes an excellent reproduction down to submicron features. You can even make a good positive image by making a replica of the replica with this material.
I have also used a product called ReproRubber, which is marketed to tool makers. This material is much cheaper than the Struers product, so may be preferable for replicating larger areas/volumes. I have done comparisons of the surface features on the replicas from this material to the original surface using SEM and quantitative surface profiling with interferometry. Comparisons were good at least to the micron level, but I did not evaluate smaller features. Hope this helps.
One disadvantage of these silicone materials is vacuum compatibility. Even moderately large replicas may outgas excessively for observation in high vacuum instruments (FESEM at 10-7 Torr), but I have not had a problem at standard SEM vacuum (10-5 Torr).
--
Larry D. Hanke, P.E. Materials Evaluation and Engineering, Inc. Practical Solutions Through Technology and Innovation http://www.mee-inc.com (763) 449-8870
} Question: Hello Listers, } I'm interested to know if anyone has experience using dental impression materials (vinyl polysiloxanes) or mold/impression brands such as Struers or GE for replicating surfaces for SEM examination. } Thanks kindly for your time and info. } } Ed Dargelis } evdargelis-at-esi-il.com } } Engineering Systems, Inc. } 3851 Exchange Ave. } Aurora, IL 60504
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 8 10:57:31 2003
Yes, I have a great deal of experience in this. What did you want to know?
Lesley Weston.
n 07/10/2003 4:12 PM, by way of MicroscopyListserver at evdargelis-at-esi-il.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 } ---------------------------------------------------------------------------- --} - } } Below is the result of your feedback form (NJZFM-ultra-55). It was submitted } by (evdargelis-at-esi-il.com) from } http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, October } 7, 2003 at 16:48:50 } --------------------------------------------------------------------------- } } Email: evdargelis-at-esi-il.com } Name: Ed Dargelis } } Organization: Engingeering Systems, Inc., 3851 Exchange Ave., Aurora, IL } 60504 } } Title-Subject: [Microscopy] MListserver: SEM Impression Materials } } Question: Hello Listers, } I'm interested to know if anyone has experience using dental impression } materials (vinyl polysiloxanes) or mold/impression brands such as Struers or } GE for replicating surfaces for SEM examination. } Thanks kindly for your time and info. } } Ed Dargelis } evdargelis-at-esi-il.com } } Engineering Systems, Inc. } 3851 Exchange Ave. } Aurora, IL 60504 } } --------------------------------------------------------------------------- }
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 8 13:35:20 2003
} I am a new user in EM and I am having a difficult time getting my Epoxy } sections to stick to the copper grids. I have tried Acetic acid washes, } sonicators, HCl wash etc. I have not tried BSA, I am waiting for the } order to come in. Does anyone have any more suggestions? } } Thank you in Advance. } Sue
Here's my methodology for collecting epoxy sections onto mesh grids and getting them to stay there. Works very well 99% of the time:
I always clean, by sonication for 30-60 seconds in dedicated clean 25ml glass beaker, my uncoated copper mesh grids (no support films, like Formvar) in the following solution:
10% concentrated HCl 20% acetone 70% distilled water
Note: Be careful mixing this up. Add HCl slowly to water, then add acetone. Store in clean capped glass bottle. pH paper tests gives pH =1.0.
After sonication in cleaning solution, rinse 2x in 99% acetone, air dry, invert beaker over clean filter paper and they will fall down as acetone rinse drys.
Always clean grids the day you section. Even after overnight, I find that some sections might tend to loosen up a bit on copper grids during staining procedures, I guess due to new oxidation layer forming within a day or two (rust never sleeps!).
I think the HCl removes oxidation layers from the grids, acetone breaks the surface tension of the aqueous solution so grids don't float, and also cleans, and facilitates quick air drying.
Also, during staining, minimize the turbulance of stains and rinses over section surfaces on grids. For that reason, I like to use the SynapTek(tm) GridStick(tm) pipet method from Ted Pella (I have no financial interest in Pella, just a satisfied user of GridStick system).
I prefer to pick up sections from the knife's water filled trough from above, rather than to slide grid underneath sections (underwater) and lift up. For me, collecting from above allows me to place a group of sections right where I want them on the grid, without them sliding off to one side or wrapping around the grid edge. Also, just as I place grid over sections, I press very slightly down, but not nearly enough to break surface of water, which resultant slight water pressure seems to push the sections onto grid bars so they stay put. Then after lifting grid off water surface, I blot the edge of the grid with a filter paper point to remove that drop of water that is hanging from the grid, air dry, or dry over slide warmer surface, but not too hot. Be sure to give adequate drying before you move on to the staining.
But I think the crucial step is the cleaning of the grids using above solution same day as they are used to collect sections. I've been using this method for many years with consistent success. Hope this helps you, and 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 } -- } Sue Tyler } Biologist } Cooperative Oxford Laboratory } Center for Coastal Environmental Health& } Biomolecular Research at Charleston ( CCHEBR) } USDOC/NOAA/NOS/NCCOS } 904 S. Morris St. } Oxford, Maryland 21654-9724 } 410-226-5193 Fax: 410- 226-5925 } Sue.Tyler-at-noaa.gov } I am a new user in EM and I am having a difficult time getting my Epoxy } sections to stick to the copper grids. I have tried Acetic acid washes, } sonicators, HCl wash etc. I have not tried BSA, I am waiting for the } order to come in. Does anyone have any more suggestions? } } Thank you in Advance. } Sue
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 8 15:41:40 2003
We are setting up to develop Kodak SO163 film for low dose cryoEM. We are push processing it, so we are developing it for 12 minutes in straight D-19, we're using a nitrogen gas burst, for 1 second every 8 seconds.
I was wondering what the life of the chemistry might be. Kodak said that we could develop 60 8x10 negatives per gallon. This works out to about 360 3 1/4 x 4 negatives. This seems like a lot of negatives to us, so we are wondering what other people are using for an exhaustion time, both in negative numbers and days in the tank.
Thanks for the help. Leslie Cummins
Leslie Gunther Cummins Analytical Imaging Facility Albert Einstein College of Medicine 1300 Morris Park Ave. Bronx, NY 10461 718-430-3547
http://www.aecom.yu.edu/aif/
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 8 19:07:02 2003
Massimo- We put a water droplet on the slide. There is a device that heats the slide up to about 45 degrees C. When the wax roll is put onto the rounded-up surface of the droplet, it gradually unfolds and stretches out to occupy the greatest possible area on the droplet. Carol Heckman (Bowling Green State University)
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From MicroscopyL-request-at-ns.microscopy.com Wed Oct 8 21:17:15 2003
On Wednesday, October 8, 2003, at 10:55 AM, Leslie Cummins wrote:
} We are setting up to develop Kodak SO163 film for low dose cryoEM. We } are push processing it, so we are developing it for 12 minutes in } straight D-19, we're using a nitrogen gas burst, for 1 second every 8 } seconds. } } I was wondering what the life of the chemistry might be. Kodak said } that we could develop 60 8x10 negatives per gallon. This works out to } about 360 3 1/4 x 4 negatives. This seems like a lot of negatives to } us, so we are wondering what other people are using for an exhaustion } time, both in negative numbers and days in the tank. } } Thanks for the help. } Dear Leslie, We've done 250 in the past, and that is our plan, if we have as many as 250 films in a short period--a week or two. If the solution is older that two weeks, we'll pitch it. You can also check by color; light brown is fresh, reddish orange is OK, and when the color intensifies to dark reddish brown, it's bad. We have covered tanks, and the developer is pretty well nitrogenated after use, but, if your tanks allow oxygen to dissolve in the developer over time, YMMV. Yours, Bill Tivol 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 8 23:43:41 2003
You can check out our website for the various Olympus trinocular ports: http://www.mvia.com/Coolpix/clpxadpt.htm#Olympus
As far as the mount that goes on top, it would be dictated by the camera you are using, but we have C-mount F-mount, bayonet mount, digital camera mounts, etc. that will all fit onto your scope.
Feel free to give me a call or email me if you need any further info.
Thanks! Jim Haley
****************************** Jim Haley Applications Engineer MVIA, Inc. 125 Sherwood Drive Monaca, PA 15061 voice: (724) 728-7493 fax: (412) 291-1709 e-mail: haley-at-mvia.com webpage: http://www.mvia.com ******************************
Philip Oshel 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 } ------------------------------------------------------------------------------- } } Micromavens, } } Does anyone have an Olympus BX51 with a phototube? We don't around } here, and the Olympus information is unclear as to: mount, C-mount or } bayonet? at the top, and if there is a projection/transfer lens } within the tube, and if so (there must be), where exactly and what } power? 1? 0.32? etc. } Thanks. } } Phil } } -- } Philip Oshel } Supervisor, BBPIC microscopy facility } Department of Animal Sciences } University of Wisconsin } 1675 Observatory Drive } Madison, WI 53706 - 1284 } voice: (608) 263-4162 } fax: (608) 262-5157 (dept. fax)
--
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 9 04:21:26 2003
Ed A couple of artefacts are worth looking out for. Delicate surface structures can sometimes be stripped from the specimen surface when the silicone replica is removed. For example, silicone replicas will strip the epicuticular wax layer from plant surfaces. When this happens, direct examination of the uncleaned negative replica in the SEM may falsely suggest that the replica has failed to record the finest structures. Similarly, an epoxy resin casting will record the fracture surface at the base of the stripped structures, and not the original outer surface structure, again suggesting poor replica performance. If this happens to you, consider the chemistry of the stripped material and consider whether it can be removed using a solvent. in the case of epicuticular waxes, I have found that fidelity of the positive replica is improved if a hot-melt method is used, because the wax embedded in the silicone is melted and displaced by hot plastic. Polycarbonate plastic (melting point ~200oC) is good for this.
A second problem can arise if the specimen surface is pitted or very rough, especially if subsurface pits open to the surface via restricted apertures smaller than the pits themselves. An example would be the open stomatal apertures of plants, but a percentage of the cavities in foamed materials might also fit this model. In this case, the silicone compound may freely enter the cavities while fluid, but once polymerized will form a solid plug that may break off at the narrowest point of the neck when the replica is stripped. This may destroy evidence of the aperture, leading to underestimates of porosity or surface roughness.
Chris
} } Email: evdargelis-at-esi-il.com } Name: Ed Dargelis } } Organization: Engingeering Systems, Inc., 3851 Exchange Ave., Aurora, IL } 60504 } } Title-Subject: [Microscopy] MListserver: SEM Impression Materials } } Question: Hello Listers, } I'm interested to know if anyone has experience using dental impression } materials (vinyl polysiloxanes) or mold/impression brands such as Struers or } GE for replicating surfaces for SEM examination. } Thanks kindly for your time and info. } Ed Dargelis } evdargelis-at-esi-il.com } } Engineering Systems, Inc. } 3851 Exchange Ave. } Aurora, IL 60504 --------------------------------------------------------------------------
========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 (0) 131 650 5554 FAX. #44 (0) 131 650 5392 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 9 07:17:36 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (nicholas.pearson-at-alcoa.com.au) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, October 9, 2003 at 00:24:31 ---------------------------------------------------------------------------
Email: nicholas.pearson-at-alcoa.com.au Name: Nick Pearson
Organization: Alcoa World Alumina
Title-Subject: [Microscopy] Video output to data projector
Question: I am trying to take the video output from a JEOL JSM 6400 and project it using a data projector (for a community display). The service engineer indicates that the signal is NTSC format, but having tried 4 projectors that are capable of detecting NTSC have not had any luck. 3 of the projectors indicate video signal not detected, and the other displays an unstable image. Has anybody else tried this on a JEOL instrument? Any suggestions?
(Another observation - Feeding the video signal to a monitor capable of displaying NTSC, the image displays momentarily (very small fraction of a second), then goes blank).
12 minutes in straight D-19 sounds like a huge push to me! On the other hand, I have no experience with cryo EM so ... Have you talked to Kodak about this? Long times in developer can lead to fogging. As to life of the chemistry in a tank. If you have a floating lid (that excludes most air) the chemistry will keep better than if the lid allows free access to air. Again, a call to Kodak is in order. As for the number of films I always stay on the conservative side, maybe 2/3 of the recommendation. D-19 is the cheapest part of the equation (animals, prep time, etc) so trying to squeez a few extra films out of a pot of developer is not the way to go. Also, D-19 does not last more than 2 years in the package on the shelf. I don't do much TEM anymore so I mix my D-19 from scratch using the individual chemicals, not the prepackaged Kodak mix. The recipe is widely published, Kodak will give it to you if you ask.
Geoff
Leslie Cummins wrote:
} Hello Listers } } We are setting up to develop Kodak SO163 film for low dose cryoEM. We } are push processing it, so we are developing it for 12 minutes in } straight D-19, we're using a nitrogen gas burst, for 1 second every 8 } seconds. } } I was wondering what the life of the chemistry might be. Kodak said } that we could develop 60 8x10 negatives per gallon. This works out to } about 360 3 1/4 x 4 negatives. This seems like a lot of negatives to } us, so we are wondering what other people are using for an exhaustion } time, both in negative numbers and days in the tank. } } Thanks for the help. } Leslie Cummins } } } Leslie Gunther Cummins } Analytical Imaging Facility } Albert Einstein College of Medicine } 1300 Morris Park Ave. } Bronx, NY 10461 } 718-430-3547 } } http://www.aecom.yu.edu/aif/ } }
-- -- ********************************************** 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 Thu Oct 9 08:33:18 2003
I've never used SO163, but with the Kodak 4489 film (yes, I still have some left over!), our rule of thumb: 100 negatives and/or 2 weeks whichever comes first. Then we discard the D-19 in tank and refresh with new. (When we make it up, it usually makes more than a tank holds, so we store the rest in a brown (light-free) bottle. For use: mix 1 part D-19:2parts H20. Hope this helps. Note: the stock solution should be good until it gets brown--it should be relatively "clear".
Peggy
Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of 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: Leslie Cummins [mailto:gunther-at-aecom.yu.edu] Sent: Wednesday, October 08, 2003 1:55 PM To: Microscopy-at-MSA.Microscopy.Com
Hello Listers
We are setting up to develop Kodak SO163 film for low dose cryoEM. We are push processing it, so we are developing it for 12 minutes in straight D-19, we're using a nitrogen gas burst, for 1 second every 8 seconds.
I was wondering what the life of the chemistry might be. Kodak said that we could develop 60 8x10 negatives per gallon. This works out to about 360 3 1/4 x 4 negatives. This seems like a lot of negatives to us, so we are wondering what other people are using for an exhaustion time, both in negative numbers and days in the tank.
Thanks for the help. Leslie Cummins
Leslie Gunther Cummins Analytical Imaging Facility Albert Einstein College of Medicine 1300 Morris Park Ave. Bronx, NY 10461 718-430-3547
http://www.aecom.yu.edu/aif/
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 9 08:52:24 2003
We are currently changing the D19 in our low-dose darkroom after 280 films are processed and this usually happens within 3 weeks of preparation.
On a side note...has anyone ever come across a batch of D19 with brown crystals? It appeared very dark once prepared and was thrown out without attempting to use it.
cheers, paul
} } } } On Wednesday, October 8, 2003, at 10:55 AM, Leslie Cummins wrote: } } } We are setting up to develop Kodak SO163 film for low dose cryoEM. } } We are push processing it, so we are developing it for 12 minutes } } in straight D-19, we're using a nitrogen gas burst, for 1 second } } every 8 seconds. } } } } I was wondering what the life of the chemistry might be. Kodak } } said that we could develop 60 8x10 negatives per gallon. This } } works out to about 360 3 1/4 x 4 negatives. This seems like a lot } } of negatives to us, so we are wondering what other people are using } } for an exhaustion time, both in negative numbers and days in the } } tank. } } } } Thanks for the help. } } } Dear Leslie, } We've done 250 in the past, and that is our plan, if we have } as many as 250 films in a short period--a week or two. If the } solution is older that two weeks, we'll pitch it. You can also } check by color; light brown is fresh, reddish orange is OK, and when } the color intensifies to dark reddish brown, it's bad. We have } covered tanks, and the developer is pretty well nitrogenated after } use, but, if your tanks allow oxygen to dissolve in the developer } over time, YMMV. } Yours, } Bill Tivol } 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
-- Paul Chipman Electron Microscopy Facility Manager Dept. of Biology, Purdue University Lilly Hall, Rm. B216 Phone: 765-494-1487 Fax:765-496-1189
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 9 08:56:20 2003
} I am a new user in EM and I am having a difficult time getting my Epoxy } sections to stick to the copper grids. I have tried Acetic acid washes, } sonicators, HCl wash etc. I have not tried BSA, I am waiting for the } order to come in. Does anyone have any more suggestions? } } Thank you in Advance. } Sue
Hi Sue, With the technique that we use, no known force in the universe will be able to remove the sections from your grids.
First of all, use clean grids. There are several ways to clean grids, but the way that I like, because it is fast, is to hold the grid in concentrated NaOH while I count to 6, and then briefly dip it repeatedly in water to rinse the grid. I do this just before I pick up the sections.
Then, most importantly, we dry down our grids in a drying oven. The oven is just hot enough to dry glassware, but not really superhot. In other words, I can put my hand on the bottom of the oven without burning it, but I really wouldn't want to keep my hand there for more than a second or so. And the temperature isn't hot enough to warp our plastic petri dishes. If it starts to warp the petri dishes, it's too hot, though it still probably won't hurt your sections. [we know that from experience] You can dry your sections there for about 5-10 minutes. But if you even forget about them overnight, it won't hurt them at all.
Then, I can assure you that your sections will NEVER NEVER NEVER wash off your grids no matter how vigorously you wash them during staining.
Garry
Garry Burgess Charge Technologist Electron Microscopy Department of Pathology Health Sciences Centre Winnipeg, Canada
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 9 08:59:43 2003
Forgive me if you've covered this possibility already but we once had some difficulty getting video out of our 6400 under similar circumstances. We eventually realised that it was because we had it synced to the line frequency (to null out the wobbles from room fields etc) and not the "standard" rate. Try toggling this feature on and off and see if it makes any difference. That is, at the bottom of the FIS-1 Functions menu (normally brought up with the F3 key), you will see the VIDO on/off item. With the scan going at TV rate scroll down and toggle the VIDO setting on and off and see if one of the settings (presumably the one that gives video operating at the conventional sync rate) results in a signal that your data projector can understand.
Good luck.
} Email: nicholas.pearson-at-alcoa.com.au } Name: Nick Pearson } } Organization: Alcoa World Alumina } } Title-Subject: [Microscopy] Video output to data projector } } Question: I am trying to take the video output from a JEOL JSM 6400 } and project it using a data projector (for a community display). } The service engineer indicates that the signal is NTSC format, but } having tried 4 projectors that are capable of detecting NTSC have } not had any luck. 3 of the projectors indicate video signal not } detected, and the other displays an unstable image. } Has anybody else tried this on a JEOL instrument? } Any suggestions? } } (Another observation - Feeding the video signal to a monitor capable } of displaying NTSC, the image displays momentarily (very small } fraction of a second), then goes blank). } } ---------------------------------------------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 9 09:05:44 2003
Forgive me if you've covered this possibility already but we once had some difficulty getting video out of our 6400 under similar circumstances. We eventually realised that it was because we had it synced to the line frequency (to null out the wobbles from room fields etc) and not the "standard" rate. Try toggling this feature on and off and see if it makes any difference. That is, at the bottom of the FIS-1 Functions menu (normally brought up with the F3 key), you will see the VIDO on/off item. With the scan going at TV rate scroll down and toggle the VIDO setting on and off and see if one of the settings (presumably the one that gives video operating at the conventional sync rate) results in a signal that your data projector can understand.
Good luck.
} Email: nicholas.pearson-at-alcoa.com.au } Name: Nick Pearson } } Organization: Alcoa World Alumina } } Title-Subject: [Microscopy] Video output to data projector } } Question: I am trying to take the video output from a JEOL JSM 6400 } and project it using a data projector (for a community display). } The service engineer indicates that the signal is NTSC format, but } having tried 4 projectors that are capable of detecting NTSC have } not had any luck. 3 of the projectors indicate video signal not } detected, and the other displays an unstable image. } Has anybody else tried this on a JEOL instrument? } Any suggestions? } } (Another observation - Feeding the video signal to a monitor capable } of displaying NTSC, the image displays momentarily (very small } fraction of a second), then goes blank). } } ---------------------------------------------------------------------------
Arthur Day, Electron Microscope Unit Phone: 61-2-9717-3457 Ansto Materials Division Fax: 61-2-9543-7179 PMB 1, Menai (Sydney), NSW, 2234 Email: ard-at-ansto.gov.au Australia www: http://www.ansto.gov.au/
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 9 11:12:54 2003
} I've recently inherited a LKB glass knife maker. I've used these beast } before, but not the Type 7801B. If anyone has a set of instructions for } this unit, would you please contact me and fax me a copy? } } Frank Karl } Degussa Corp. } frank.karl-at-degussa.com } Fax 330-668-3846 ----- Frank, I have a copy of the instructions and will fax them shortly.
Patricia Stranen Connelly The University of Pennsylvania Department of Biology Philadelphia, PA 91904-6018 215-898-7145 psconnel-at-sas.upenn.edu
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 9 11:40:42 2003
We use 2 parts water, 1 part full strength D-19 and develop using 1 second nitrogen burst every 8 seconds for 10 minutes. With this recipe we get about 400 micrographs per gal. Incidentally, we have more problems with cross contamination of solutions than chemical exhaustion. Therefore we test the developer solution before each run using a test sheet of kodabromide print paper by immersing for 2 minutes to check the paper then lights on and back into the developer. The paper should turn black relatively quickly if the solution is good.
Fran Laabs
Ames Lab, ISU
On Wednesday, October 8, 2003, at 10:55 AM, Leslie Cummins wrote:
We are setting up to develop Kodak SO163 film for low dose cryoEM. We are push processing it, so we are developing it for 12 minutes in straight D-19, we're using a nitrogen gas burst, for 1 second every 8 seconds.
I was wondering what the life of the chemistry might be. Kodak said that we could develop 60 8x10 negatives per gallon. This works out to about 360 3 1/4 x 4 negatives. This seems like a lot of negatives to us, so we are wondering what other people are using for an exhaustion time, both in negative numbers and days in the tank.
Thanks for the help.
Dear Leslie, We've done 250 in the past, and that is our plan, if we have as many as 250 films in a short period--a week or two. If the solution is older that two weeks, we'll pitch it. You can also check by color; light brown is fresh, reddish orange is OK, and when the color intensifies to dark reddish brown, it's bad. We have covered tanks, and the developer is pretty well nitrogenated after use, but, if your tanks allow oxygen to dissolve in the developer over time, YMMV. Yours, Bill Tivol 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 Thu Oct 9 11:39:43 2003
I have used brown developer made with brown crystals before in my home darkroom (not in the lab!) and have had satisfactory results. As I recall, the developers were Microdol-X for film and Dektol for paper. This doesn't mean I would ever risk anyone else's films in it, but for the replaceable stuff I was doing at the time, it was worth the experiment.
Randy
Randy Tindall EM Specialist Electron Microscopy Core---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.biotech.missouri.edu/emc/
-----Original Message----- } From: Paul Chipman [mailto:paulrc-at-bilbo.bio.purdue.edu] Sent: Thursday, October 09, 2003 8:51 AM To: microscopy-at-msa.microscopy.com
Hi Leslie,
We are currently changing the D19 in our low-dose darkroom after 280 films are processed and this usually happens within 3 weeks of preparation.
On a side note...has anyone ever come across a batch of D19 with brown crystals? It appeared very dark once prepared and was thrown out without attempting to use it.
cheers, paul
} } } } On Wednesday, October 8, 2003, at 10:55 AM, Leslie Cummins wrote: } } } We are setting up to develop Kodak SO163 film for low dose cryoEM. } } We are push processing it, so we are developing it for 12 minutes } } in straight D-19, we're using a nitrogen gas burst, for 1 second } } every 8 seconds. } } } } I was wondering what the life of the chemistry might be. Kodak } } said that we could develop 60 8x10 negatives per gallon. This } } works out to about 360 3 1/4 x 4 negatives. This seems like a lot } } of negatives to us, so we are wondering what other people are using } } for an exhaustion time, both in negative numbers and days in the } } tank. } } } } Thanks for the help. } } } Dear Leslie, } We've done 250 in the past, and that is our plan, if we have } as many as 250 films in a short period--a week or two. If the } solution is older that two weeks, we'll pitch it. You can also } check by color; light brown is fresh, reddish orange is OK, and when } the color intensifies to dark reddish brown, it's bad. We have } covered tanks, and the developer is pretty well nitrogenated after } use, but, if your tanks allow oxygen to dissolve in the developer } over time, YMMV. } Yours, } Bill Tivol } 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
-- Paul Chipman Electron Microscopy Facility Manager Dept. of Biology, Purdue University Lilly Hall, Rm. B216 Phone: 765-494-1487 Fax:765-496-1189
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 9 12:56:05 2003
I just wash my grids in 100% Acetone and then water to rinse and let them dry off over a hot plate.... has worked for me for the past 4 years.. no problems...
Eric
================================================= } Hi Sue, } } You wrote: } } } I am a new user in EM and I am having a difficult time getting my Epoxy } } sections to stick to the copper grids. I have tried Acetic acid washes, } } sonicators, HCl wash etc. I have not tried BSA, I am waiting for the } } order to come in. Does anyone have any more suggestions?
} Hi Sue, } With the technique that we use, no known force in the universe will be able } to remove the sections from your grids. } } First of all, use clean grids. There are several ways to clean grids, but } the way that I like, because it is fast, is to hold the grid in concentrated } NaOH while I count to 6, and then briefly dip it repeatedly in water to } rinse the grid. I do this just before I pick up the sections. } } Then, most importantly, we dry down our grids in a drying oven. The oven is } just hot enough to dry glassware, but not really superhot. In other words, } I can put my hand on the bottom of the oven without burning it, but I really } wouldn't want to keep my hand there for more than a second or so. And the } temperature isn't hot enough to warp our plastic petri dishes. If it starts } to warp the petri dishes, it's too hot, though it still probably won't hurt } your sections. [we know that from experience] You can dry your sections } there for about 5-10 minutes. But if you even forget about them overnight, } it won't hurt them at all. } } Then, I can assure you that your sections will NEVER NEVER NEVER wash off } your grids no matter how vigorously you wash them during staining. } } Garry } } Garry Burgess } Charge Technologist } Electron Microscopy } Department of Pathology } Health Sciences Centre } Winnipeg, Canada }
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 9 18:49:52 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dsd-at-ogpnet.com; d_scott_davis-at-yahoo.com) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Thursday, October 9, 2003 at 13:57:46 ---------------------------------------------------------------------------
Email: dsd-at-ogpnet.com; d_scott_davis-at-yahoo.com Name: Scott Davis
Organization: Optical Gaging Products, Inc.
Education: Graduate College
Location: Rochester, NY 14621 USA
Question: Where would you suggest that I look for information on the current state of the art in X-ray Photoelectron Spectroscopy for use in semiconductor dielectric thickness and nitrogen doping concentration measurement? Is NIST current or lagging? How about SEMATECH? Or is this technology really in the hands of metrology companies and therefore proprietary information?
Basically, the life of unused (even diluted) developer is quite long (without air). The clock starts ticking as soon as you develop even one piece of the film (similar but less dramatic for paper developers). The amount of film company recommends to develop usually mean that you supposed to develop all those films in relatively short period of time. Classical example is quite popular in amateur photography D-76 developer. In 1 liter of the fresh developer you could develop about 5 rolls of 35 mm film, but developer will die in about 4-5 hours does not matter how many films you manage to develop, so hurry up! Similarly, it's true for D-19 especially non-diluted. It will not die in few hours, but perhaps two weeks. The exposure to air is very critical here also. So, the bottom line here is: if you need to develop a lot of films in quite short period of time - you may do it to extend company's recommendation. As soon as you start use developer, it will gradually lost capacity in the matter of few weeks and will die even if you just developed 10 films. Note: this does not applied to the fixer - as long as you do not contaminate it with developer, it will works to the end of capacity, does not matter how long it will take (6 mo for sure and even longer). Fixer will die soon if you contaminate it with developer. By the way - if you are using those test drops to check fixer - they are too sensitive, you may use fixer much longer and for more films without any harm to the film. Experimentally I determined that when those "drops" indicate, the fixer is bad, it actually used about 50% of capacity. I hope it helps. Sergey
At 06:11 AM 10/9/2003, 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 33-089 Los Angeles, CA 90095
While I don't use SO163 film or compatible developers, I disagree in general about the life of film developer mix. The film developer is the most important step in processing silver halide media. At least, this is how I see it.
I use (or would have used) fresh developer for each batch of film that I developed. And Kodak explicitly details (as do others) how many rolls or sheets of film can be processed with a fresh batch before refreshing or dumping a pot of developer. I never refreshed developer. The results are just not the same as with a fresh batch.
It may be that this TEM film is pathetically ignorant of basic photo chemistry. However, for such a small cost of chemicals, why take the chance? If the shots are worth shooting, they are worth proper processing. That means new chemicals. And, these chemicals must be at the correct temperature to be consistent and effective. I've seen too many instances of no temperature control. Consequently, the results vary wildly. Duh.
Furthermore, I dumped stop bath and fixer routinely. Why cut corners after so much work on the instrument?
gary g.
At 05:12 PM 10/9/2003, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 9 22:51:18 2003
Garry You are right and aren't. Kodak spent a lot of time (and perhaps money) to develop chemical formulations, which are stable over some period of time being in use. It, actually, made a revolution in the photographic process: all C-41 process machines in our drug-stores used the solutions, which stable for quite a while. It dramatically reduces the cost of developing and maintenance. Similarly they invented "long-play" solutions to develop X-rays films in the hospitals (we do have such machine - technician changed solutions ones a month). Because in TEM it's just impossible to use fresh developer every time you need to develop a few films and Kodak suggest to use "used" developer over some period of time, I suspect D-19 has "extended life" formulation as well. Based on my personal experience, it's clear to me that diluted solution of D-19 (used or non-used) deliver about the same results for at least 3 weeks if properly stored without exposure to air. Usually we develop about 100 films in 1.5 liter. I think, the decent quality of the TEM film developed in used developer is also because of special film formulation (bye-bye old 4489!). It's called "electron microscopy film" and I assume it separates this film from other type of the films. As for price for chemicals - please count your technician time to make fresh solutions 3 times a day and utilize used ones, it's not cheap as you think. On the positive side, I do agree with you that it's good idea in general to use fresh components everywhere: in photography, biochemistry, cooking etc. Sergey
P.S. You don't need to change Stop-bath and fixer so often. I do believe, that Kodak suggests that stop-bath solution is good for 1000+ films... The things about fixer - it could not spoil your film as long as you careful do not mix it with developer. You may left film in the fixer for hours and it does not harm film. You need to wash it after all by the way. The bottom line here is that you may extend fixation time if your fixer start working slowly saving money and doing good for environment (less chemical waste).
At 07:46 PM 10/9/2003, you wrote: } While I don't use SO163 film or compatible developers, } I disagree in general about the life of film developer mix. } The film developer is the most important step in } processing silver halide media. At least, this is } how I see it. } } I use (or would have used) fresh developer for each } batch of film that I developed. And Kodak explicitly } details (as do others) how many rolls or sheets of film } can be processed with a fresh batch before refreshing } or dumping a pot of developer. I never refreshed } developer. The results are just not the same as with } a fresh batch. } } It may be that this TEM film is pathetically ignorant } of basic photo chemistry. However, for such a small } cost of chemicals, why take the chance? If the shots } are worth shooting, they are worth proper processing. } That means new chemicals. And, these chemicals must } be at the correct temperature to be consistent and } effective. I've seen too many instances of no temperature } control. Consequently, the results vary wildly. Duh. } } Furthermore, I dumped stop bath and fixer routinely. } Why cut corners after so much work on the instrument? } } gary g. } } } } At 05:12 PM 10/9/2003, 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 33-089 Los Angeles, CA 90095
The FocusOnMicroscopy FOM2004 international conference, announced to take place in Singapore, will now be held in Philadelphia, PA, USA, April 4 to April 7. Various organizational issues and travel concerns made this move necessary.
The 2004 meeting will be hosted by Drexel University, School of Biomedical Engineering, Science and Health Systems. The conference and exhibition will be located at the Sheraton University City Hotel, central on the Philadelphia campus. As the next in a series of unique interdisciplinary meetings on advanced multidimensional light microscopy and image processing, the 2004 meeting will pay special attention to the conjunction of multidimensional microscopies with the areas of bioinformatics, bio-nanotechnology and bioengineering.
Deadline for the submission of abstracts will be January 30, 2004. Further information on location, registration, and abstract submission will soon be made available at the Focus on Microscopy webpage: www.focusonmicroscopy.org. We invite you to participate in this conference on behalf the FOM 2004 organizing committee: A. Kriete, Pittsburgh; F. Brakenhoff, Amsterdam; P.C. Cheng, Buffalo; Banu Onaral, Philadelphia. -- +-----------------------------------------+ Prof. Dr. G.J. Brakenhoff University of Amsterdam Institute for Molecular Cell Biology Section Molecular Cytology Kruislaan 316 1098 SM Amsterdam, The Netherlands
Sergey Sodium thiosulphate has a mild solvent effect on silver, and this is accelerated at low pH in acid and hardening fixer formulations. The consequences are negligible during standard fixing times for film or paper, but prolonged over-fixing will produce noticeable bleaching, reducing overall negative or print density and bleaching out low density areas so that shadow detail in negatives and highlight detail in prints are lost.
An old rule of thumb to determine the required fix time for EM film in old or partially-used fixer is twice the time taken to clear the film. Another rule of thumb is to discard the fixer when the clearing time is twice that for a fresh solution. Development is stopped very quickly in a fix bath unless it is completely exhausted, and it is perfectly safe to inspect the negatives as they clear in safelight or even in subdued white light. The most serious downside of stretching fixer life to the limit is that it increases the risk of silver staining of the negatives, and may reduce their archival permanence.
I gave a formula for D19 on this list some months back. Here it is again, derived from British Journal of Photography almanac.
However, other sources quote markedly different proportions of the ingredients, e.g. this one from Leica Users group http://mejac.palo-alto.ca.us/leica-users/v03/msg13172.html D19: water 500 cc Metol 2.0 grams sodium sulfite, desc. 90.0 grams Hydroquinone 8.0 grams sodium carbonate, monohyd. 52.5 grams potassium bromide 5 grams cold water to make 1.0 liter and this one from http://astro.umsystem.edu/apml/ARCHIVES/MAR01/msg01118.html You can make D19 yourself by using its formula: Metol 2 g } } Sodium Sulfite 90 g } } Hydroquinone 8 g } } Sodium Carbonate (anhydrous) 45 g } } Potassium Bromide 5 g } } Water 1 liter
What is the official formula of Kodak D19 as currently marketed. Anyone have authoritative information?
Dr. Chris Jeffree University of Edinburgh Biological Sciences EM Faciility
----- Original Message ----- } From: "Sergey Ryazantsev" {sryazant-at-ucla.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Friday, October 10, 2003 4:49 AM
The Analytical Instrumentation Facility at NC State University has an opening for a staff member. Please respond directly to Prof. Phil Russell. Do not respond to this email.
NC State University Analytical Instrumentation Facility SEM Laboratory Supervisor
The North Carolina State University Analytical Instrumentation Facility (AIF) is seeking a qualified person to join AIF's professional team of microscopists and microanalysts as the Laboratory Supervisor of the Variable Pressure SEM laboratory. Variable Pressure SEM Laboratory Supervisor is a full time permanent research staff position. Duties and responsibilities include: Management of AIF's Variable Pressure SEM/EDS laboratory; operation and maintenance of Variable Pressure SEM instrumentation and related sample preparation and analysis equipment; scheduling of access to and oversight of the above instrumentation; user training and assistance; assistance with the teaching of electron microscopy laboratory classes; and analysis of a wide variety of samples including development of new analytical techniques when applicable. Duties also include responsibility for providing back up support for the FESEM and SPM laboratories. Qualifications must include a BS or higher in a Materials Science related discipline. Required skills must include: one or more years HANDS ON experience with SEM and related techniques and accessories (e.g. EDS, specimen prerparation and other associated analytical tools) and a record of relavent technical publications and presentations. Preferred qualifications include: MS or higher degree or equivalent, teaching of SEM and SEM user training; familiarity with modern electronics; experience with vacuum systems and some experience with SPM and/or surface analytical techniques. Opportunities for professional growth include opportunities to learn new techniques including STEM, FIB, surface analysis techniques and advanced sample preparation techniques.
Please send resume and three letters of reference to: Phillip E. Russell, Director; Analytical Instrumentation Facility; North Carolina State University; Box 7531, Room 318A EGRC; 2410 Campus Shore Drive; Raleigh, NC 27695-7531 or email prussell-at-ncsu.edu.
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From MicroscopyL-request-at-ns.microscopy.com Fri Oct 10 15:04:16 2003
From Kodak publication J-1, "Processing Chemicals and Formulas for Black and White Photography"
D-19
Water at 50 degrees C/ 125 degrees F 500 ml Elon (Kodak's name for Metol*) 2.0 grams Sodium sulfite, anhydrous 90.0 grams Hydroquinone 8.0 grams Sodium carbonate, monohydrated 52.5 grams Potassium bromide, anhydrous 5.0 grams water to make one liter.
* metol is p-methylamino-phenol sulfate, C.A.S 55-55-0
Geoff
-- -- ********************************************** 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 10 18:38:38 2003
On Thursday, October 9, 2003, at 06:50 AM, Paul Chipman wrote:
} On a side note...has anyone ever come across a batch of D19 with brown } crystals? It appeared very dark once prepared and was thrown out } without attempting to use it.
Dear Paul, Yes, the xtals will turn yellow, then brown. They have been oxidized, and discarding the package is best. As Geoff McAuliffe said, D-19 is the cheapest item in the procedure. If, however, you have no other D-19 available, and the solution is not dark brown when mixed, you can safely develop some film with it in a pinch. Yours, Bill Tivol 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 10 22:40:05 2003
Chris Thanks for your message. It forced me to refresh my memory in chemistry. The process of dissolving of metal silver is reduction. Metal Ag in the film is already reduced in compare with silver halide. The conditions in the fixer are again reducing, so there is no chemical direct way to reduce metal silver in the fixer. It meant that directly sodium thiosulphate could not dissolve the metal silver in the film but silver halide. There are couple of things you need to keep in mind: if for some reason silver is oxidized, then you could reduce it with sodium thiosulphate and therefore "dissolve it". Because general reducing conditions in the fixer, oxygen from air may not be effective. You need to add something like bleach into the fixer to oxidize silver and then you may reduce it with sodium thiosulphate. Another thing comes to my mind is chlor gas. So, you may keep film in the fixer for quite while and silver will not dissolved (if only you will not add the bleach). Professional photographers usually do not recommend to keep film in fixer for so long for another reason: sodium thiosulphate is slowly decomposing with creation of elementary sulphur, which colored film in yellowish color. If you will keep your film in the fixer for couple of days, elementary sulfur will be deposited in the gelatin layer, which made film yellowish irreversibly. Similar thing is happening when you do not wash film well after fixer - the residue of sodium thiosulphate will decompose with sulphur creation. So, it's good practice to ensure your film is washed well after fixer. Have a great wekend, Sergey
At 03:08 AM 10/10/2003, 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 33-089 Los Angeles, CA 90095
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Lithpingthnake-at-aol.com) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Saturday, October 11, 2003 at 08:43:43 ---------------------------------------------------------------------------
Email: Lithpingthnake-at-aol.com Name: Henry Behrens
Organization: N.C.C. Long Island N.Y.
Education: Undergraduate College
Location: Garden City,N.Y. Nassau
Question: I am looking for a resource that will help me understand and get supplies related to.Can you help? Thankyou for your attention.
We had though quite good results with CD31 and vWF in thick sections and whole mounts, as sec. antibody one of the Alexa's from Probes. But what thickness are you talking about? How about inserting eGFP?
Sven _______________________________________________________________ Sven Terclavers LM/CLSM Research Assistent Center for Transgene Technology & Gene Therapy Campus Gasthuisberg O&N Herestraat 49 3000 Leuven Belgium Tel.: +32 (0)16 34 63 71 Fax: +32 (0)16 34 59 90 Email: Sven.Terclavers-at-med.kuleuven.ac.be Web: http://www.kuleuven.ac.be/mcm & http://browse.to/microscopy (only intern) _______________________________________________________________
-------Original Message-------
} From: Michael Herron
I would like suggestions for a reasonably bright marker for mouse endothelium viewed in thick section confocal.
We have tried CD31, Ulex and von Willebrand's factor, and they were all fairly worthless.
Suggestions?!
Thanks, Mike
---
Michael J. Herron, U of MN, Dept. of Entomology herro001-at-umn.edu 612-624-3688 (office) 612-625-5299 (FAX)
From MicroscopyL-request-at-ns.microscopy.com Sun Oct 12 16:59:26 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (gilbert.scheffer-at-wanadoo.fr) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Sunday, October 12, 2003 at 10:47:53 ---------------------------------------------------------------------------
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sergey-at-seas.ucla.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Friday, October 10, 2003 at 20:16:22 ---------------------------------------------------------------------------
Dear all, What books are you using in the undergrad teaching of electron microscopy? I am in a materials department so we are most concerned with EM of inorganic materials, although polymers might be worth covering too. Also, I do mostly TEM, but I do also talk more generally about SEM and other electron beam instruments too. Does anyone have particular recommendations for textbooks for the undergraduates in this area, they will be in the fifth semester (3rd year, 1st term)?
Much as I like Williams and Carter, this is still a bit big and detailed for the target audience. Mike Loretto's book used to be quite good for this sort of thing, but times have moved on. Maybe, there are newer books on the market that cover the target audience better, with some covereage of recent developments.
Thanks in advance for any useful advice you can give.
-- Ian MacLaren Technische Universität Darmstadt Material- und Geowissenschaften Petersenstr. 23 64287 Darmstadt Germany http://www.tu-darmstadt.de/fb/ms/fg/sf/projekte/maclaren-Dateien/maclaren.html
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 13 04:16:53 2003
Sergey I don't pretend to be able to account for this effect, but it happens anyway. Try this simple test: stand an unwanted negative on edge with lower half immersed in a bath of your favourite acid or rapid fixer overnight. Wash and dry. Compare density of immersed and non-immersed areas.
See quotes below from Grant Haist (1979) Modern Photographic Processing; John Wiley & Sons, Inc.; New York; Vol. 1; pps. 564-565 “Another danger of long fixing bath immersions is the direct attack on the silver image by the combination of oxygen and the acid fixing bath. It is believed that oxygen from the air dissolves in the fixing bath and attacks the very finely divided silver particles of the image. Oxygen converts these metallic silver particles to silver ions by removing electrons. The silver ions are then complexed by the thiosulfate and removed, resulting in the loss of image silver. Fine-grained films and paper images are especially susceptible to image reduction by acid thiosulfate solutions.” “The extent of the reducing effect of fixing baths on the silver image during the process of fixation is greater than has been generally supposed. This attack does not occur in alkaline thiosulfate solutions" Best wishes Chris
On 10 Oct 03, at 20:38, 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 } ---------------------------------------------------------------------- } --------- } } Chris } Thanks for your message. It forced me to refresh my memory in } chemistry. The process of dissolving of metal silver is reduction. } Metal Ag in the film is already reduced in compare with silver halide. } The conditions in the fixer are again reducing, so there is no } chemical direct way to reduce metal silver in the fixer. It meant that } directly sodium thiosulphate could not dissolve the metal silver in } the film but silver halide. There are couple of things you need to } keep in mind: if for some reason silver is oxidized, then you could } reduce it with sodium thiosulphate and therefore "dissolve it". } Because general reducing conditions in the fixer, oxygen from air may } not be effective. You need to add something like bleach into the } fixer to oxidize silver and then you may reduce it with sodium } thiosulphate. Another thing comes to my mind is chlor gas. So, you } may keep film in the fixer for quite while and silver will not } dissolved (if only you will not add the bleach). Professional } photographers usually do not recommend to keep film in fixer for so } long for another reason: sodium thiosulphate is slowly decomposing } with creation of elementary sulphur, which colored film in yellowish } color. If you will keep your film in the fixer for couple of days, } elementary sulfur will be deposited in the gelatin layer, which made } film yellowish irreversibly. Similar thing is happening when you do } not wash film well after fixer - the residue of sodium thiosulphate } will decompose with sulphur creation. So, it's good practice to ensure } your film is washed well after fixer. Have a great wekend, Sergey } } At 03:08 AM 10/10/2003, 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 } } --------------------------------------------------------------------- } } ---------- } } } } Sergey } } Sodium thiosulphate has a mild solvent effect on silver, and this is } } accelerated at low pH in acid and hardening fixer formulations. The } } consequences are negligible during standard fixing times for film or } } paper, but prolonged over-fixing will produce noticeable bleaching, } } reducing overall negative or print density and bleaching out low } } density areas so that shadow detail in negatives and highlight detail } } in prints are lost. } } } } An old rule of thumb to determine the required fix time for EM film } } in old or partially-used fixer is twice the time taken to clear the } } film. Another rule of thumb is to discard the fixer when the clearing } } time is twice that for a fresh solution. Development is stopped very } } quickly in a fix bath unless it is completely exhausted, and it is } } perfectly safe to inspect the negatives as they clear in safelight or } } even in subdued white light. The most serious downside of stretching } } fixer life to the limit is that it increases the risk of silver } } staining of the negatives, and may reduce their archival permanence. } } } } I gave a formula for D19 on this list some months back. Here it is } } again, derived from British Journal of Photography almanac. } } } } Metol 2.2g, Hydroquinone 8.8g, Sodium sulphite 144g, Sodium carbonate } } 130g Potassium bromide 4g, H2O to 1litre } } } } However, other sources quote markedly different proportions of the } } ingredients, e.g. this one from Leica Users group } } http://mejac.palo-alto.ca.us/leica-users/v03/msg13172.html D19: water } } 500 cc Metol 2.0 grams sodium sulfite, desc. 90.0 grams Hydroquinone } } 8.0 grams sodium carbonate, monohyd. 52.5 grams potassium bromide 5 } } grams cold water to make 1.0 liter and this one from } } http://astro.umsystem.edu/apml/ARCHIVES/MAR01/msg01118.html You can } } make D19 yourself by using its formula: Metol 2 g } } Sodium Sulfite } } 90 g } } Hydroquinone 8 g } } Sodium Carbonate (anhydrous) 45 g } } } } Potassium Bromide 5 g } } Water 1 liter } } } } What is the official formula of Kodak D19 as currently marketed. } } Anyone have authoritative information? } } } } } } Dr. Chris Jeffree } } University of Edinburgh } } Biological Sciences EM Faciility } } } } ----- Original Message ----- } } } From: "Sergey Ryazantsev" {sryazant-at-ucla.edu} } } To: {Microscopy-at-sparc5.microscopy.com} } } Sent: Friday, October 10, 2003 4:49 AM } } Subject: [Microscopy] Re: SO163 Development } } } } } } } } } } } } } ------------------------------------------------------------------ } } } -- } } ---------- } } } 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 } } } ------------------------------------------------------------------ } } } -- } } ----------- } } } } } } Garry } } } You are right and aren't. Kodak spent a lot of time (and perhaps } } money) to } } } develop chemical formulations, which are stable over some period } } } of } } time } } } being in use. It, actually, made a revolution in the photographic } } process: } } } all C-41 process machines in our drug-stores used the solutions, } } which } } } stable for quite a while. It dramatically reduces the cost of } } developing } } } and maintenance. Similarly they invented "long-play" solutions to } } develop } } } X-rays films in the hospitals (we do have such machine - } } } technician } } changed } } } solutions ones a month). Because in TEM it's just impossible to } } } use } } fresh } } } developer every time you need to develop a few films and Kodak } } suggest to } } } use "used" developer over some period of time, I suspect D-19 has } } "extended } } } life" formulation as well. Based on my personal experience, it's } } clear to } } } me that diluted solution of D-19 (used or non-used) deliver about } } the same } } } results for at least 3 weeks if properly stored without exposure } } } to air. Usually we develop about 100 films in 1.5 liter. I } } } think, the } } decent } } } quality of the TEM film developed in used developer is also } } } because } } of } } } special film formulation (bye-bye old 4489!). It's called } } } "electron microscopy film" and I assume it separates this film } } } from other type } } of the } } } films. As for price for chemicals - please count your technician } } time to } } } make fresh solutions 3 times a day and utilize used ones, it's not } } cheap as } } } you think. On the positive side, I do agree with you that it's } } good idea } } } in general to use fresh components everywhere: in photography, } } } biochemistry, cooking etc. Sergey } } } } } } P.S. You don't need to change Stop-bath and fixer so often. I do } } believe, } } } that Kodak suggests that stop-bath solution is good for 1000+ } } films... The } } } things about fixer - it could not spoil your film as long as you } } careful do } } } not mix it with developer. You may left film in the fixer for } } } hours } } and it } } } does not harm film. You need to wash it after all by the way. } } } The } } bottom } } } line here is that you may extend fixation time if your fixer } } } start } } working } } } slowly saving money and doing good for environment (less chemical } } waste). } } } } } } } } } At 07:46 PM 10/9/2003, you wrote: } } } } While I don't use SO163 film or compatible developers, } } } } I disagree in general about the life of film developer mix. } } } } The film developer is the most important step in } } } } processing silver halide media. At least, this is } } } } how I see it. } } } } } } } } I use (or would have used) fresh developer for each } } } } batch of film that I developed. And Kodak explicitly } } } } details (as do others) how many rolls or sheets of film } } } } can be processed with a fresh batch before refreshing } } } } or dumping a pot of developer. I never refreshed } } } } developer. The results are just not the same as with } } } } a fresh batch. } } } } } } } } It may be that this TEM film is pathetically ignorant } } } } of basic photo chemistry. However, for such a small } } } } cost of chemicals, why take the chance? If the shots } } } } are worth shooting, they are worth proper processing. } } } } That means new chemicals. And, these chemicals must } } } } be at the correct temperature to be consistent and } } } } effective. I've seen too many instances of no temperature } } } } control. Consequently, the results vary wildly. Duh. } } } } } } } } Furthermore, I dumped stop bath and fixer routinely. } } } } Why cut corners after so much work on the instrument? } } } } } } } } gary g. } } } } } } } } } } } } } } } } At 05:12 PM 10/9/2003, 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 } } } } } } } ------------------------------------------------------------------ } } } } -- } } ----------- } } } } } } } } } } Basically, the life of unused (even diluted) developer is quite } } long } } } } } (without air). The clock starts ticking as soon as you develop } } even one } } } } } piece of the film (similar but less dramatic for paper } } developers). The } } } } } amount of film company recommends to develop usually mean that } } } } } you supposed to develop all those films in relatively short } } } } } period of time. Classical example is quite popular in amateur } } } } } photography } } D-76 } } } } } developer. In 1 liter of the fresh developer you could develop } } about 5 } } } } } rolls of 35 mm film, but developer will die in about 4-5 hours } } does not } } } } } matter how many films you manage to develop, so hurry up! } } Similarly, } } } } } it's true for D-19 especially non-diluted. It will not die in } } } } } few } } hours, } } } } } but perhaps two weeks. The exposure to air is very critical } } } } } here also. So, the bottom line here is: if you need to develop } } } } } a lot } } of films } } } } } in quite short period of time - you may do it to extend } } } } } company's recommendation. As soon as you start use developer, it } } } } } will } } gradually } } } } } lost capacity in the matter of few weeks and will die even if } } } } } you } } just } } } } } developed 10 films. Note: this does not applied to the fixer - } } } } } as } } long } } } } } as you do not contaminate it with developer, it will works to } } } } } the } } end of } } } } } capacity, does not matter how long it will take (6 mo for sure } } } } } and } } even } } } } } longer). Fixer will die soon if you contaminate it with } } developer. By } } } } } the way - if you are using those test drops to check fixer - } } } } } they } } are too } } } } } sensitive, you may use fixer much longer and for more films } } without any } } } } } harm to the film. Experimentally I determined that when those } } "drops" } } } } } indicate, the fixer is bad, it actually used about 50% of } } capacity. I } } } } } hope it helps. Sergey } } } } } } } } } } } } } } } At 06:11 AM 10/9/2003, 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 Leslie: } } } } } } } } } } } } 12 minutes in straight D-19 sounds like a huge push to me! } } } } } } On } } the } } } } } } other hand, I have no experience with cryo EM so ... Have you } } talked to } } } } } } Kodak about this? Long times in developer can lead to fogging. } } } } } } As to life of the chemistry in a tank. If you have a } } } } } } floating } } lid } } } } } } (that excludes most air) the chemistry will keep better than } } } } } } if } } the } } } } } } lid allows free access to air. Again, a call to Kodak is in } } order. As } } } } } } for the number of films I always stay on the conservative } } } } } } side, } } maybe } } } } } } 2/3 of the recommendation. } } } } } } D-19 is the cheapest part of the equation (animals, prep } } time, etc) } } } } } } so trying to squeez a few extra films out of a pot of } } } } } } developer } } is not } } } } } } the way to go. Also, D-19 does not last more than 2 years in } } } } } } the package on the shelf. I don't do much TEM anymore so I mix } } } } } } my } } D-19 from } } } } } } scratch using the individual chemicals, not the prepackaged } } Kodak mix. } } } } } } The recipe is widely published, Kodak will give it to you if } } } } } } you } } ask. } } } } } } } } } } } } Geoff } } } } } } } } } } } } Leslie Cummins wrote: } } } } } } } } } } } } } Hello Listers } } } } } } } } } } } } } } We are setting up to develop Kodak SO163 film for low dose } } cryoEM. We } } } } } } } are push processing it, so we are developing it for 12 minutes } } in } } } } } } } straight D-19, we're using a nitrogen gas burst, for 1 second } } every 8 seconds. } } } } } } } } } } } } } } I was wondering what the life of the chemistry might be. } } } } } } } Kodak } } said } } } } } } } that we could develop 60 8x10 negatives per gallon. This } } } } } } } works } } out to } } } } } } } about 360 3 1/4 x 4 negatives. This seems like a lot of } } negatives to } } } } } } } us, so we are wondering what other people are using for an } } exhaustion } } } } } } } time, both in negative numbers and days in the tank. } } } } } } } } } } } } } } Thanks for the help. } } } } } } } Leslie Cummins } } } } } } } } } } } } } } } } } } } } } Leslie Gunther Cummins } } } } } } } Analytical Imaging Facility } } } } } } } Albert Einstein College of Medicine } } } } } } } 1300 Morris Park Ave. } } } } } } } Bronx, NY 10461 } } } } } } } 718-430-3547 } } } } } } } } } } } } } } http://www.aecom.yu.edu/aif/ } } } } } } } } } } } } -- } } } } } } -- } } } } } } ********************************************** } } } } } } 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 } } } } } } ********************************************** } } } } } } } } } } } } } } } } _____________________________________ } } } } } } } } } } Sergey Ryazantsev Ph. D. } } } } } Electron Microscopy } } } } } UCLA School of Medicine } } } } } Department of Biological Chemistry } } } } } 10833 Le Conte Ave, Room 33-089 } } } } } Los Angeles, CA 90095 } } } } } } } } } } Phone: (310) 825-1144 (office) } } } } } (310) 206-1029 (Lab) } } } } } FAX (departmental): (310) 206-5272 } } } } } mailto:sryazant-at-ucla.edu } } } } } } } } } } } } } } } } } } } } _____________________________________ } } } } } } Sergey Ryazantsev Ph. D. } } } Electron Microscopy } } } UCLA School of Medicine } } } Department of Biological Chemistry } } } 10833 Le Conte Ave, Room 33-089 } } } Los Angeles, CA 90095 } } } } } } Phone: (310) 825-1144 (office) } } } (310) 206-1029 (Lab) } } } FAX (departmental): (310) 206-5272 } } } mailto:sryazant-at-ucla.edu } } } } } } } } } } } } } } } } ------- End of forwarded message ------- } } ========================================== } } Dr. Chris Jeffree } } University of Edinburgh } } BIOSEM - Biological Sciences Electron Microscope Facility } } Institute of Cell and Molecular Biology } } Daniel Rutherford Building } } King's Buildings, Mayfield Road } } EDINBURGH, EH9 3JH, Scotland, UK } } Tel. #44 (0) 131 650 5554 } } FAX. #44 (0) 131 650 5392 } } Mobile 07710 585 401 } } email c.jeffree-at-ed.ac.uk } } ========================================= } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } 10833 Le Conte Ave, Room 33-089 } Los Angeles, CA 90095 } } Phone: (310) 825-1144 (office) } (310) 206-1029 (Lab) } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu } } } }
========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 (0) 131 650 5554 FAX. #44 (0) 131 650 5392 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 13 09:51:00 2003
I'm seeking a "lamp housing mirror" for a Zeiss Axiomat microscope or information on someone who may have one. Please contact me off line if you can help.
Thanks,
Owen Mills
Owen P. Mills Electron Optics Engineer Materials Science & Engineering Michigan Technological University Rm 512 M&M Bldg. Houghton, MI 49931 PH 906-369-1875 FAX 906-487-2934 mailto:opmills-at-mtu.edu http://www.mm.mtu.edu/~opmills
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 13 12:45:18 2003
There is an older book by Ian Watt. I don't know whether it is still in print. Carol Heckman
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From MicroscopyL-request-at-ns.microscopy.com Mon Oct 13 15:37:57 2003
I am attempting to ultracryosection a cell monolayer grown on a filter. I find the filter resists infiltration with sucrose/PVP, and shatters when I try sectioning. I have tried varying the cutting temperature and speed to no avail. I wonder if anyone in the field has successfully sectioned cells grown on a filter for cryo-immunoEM, and if so whether you would be willing to share your techniques (type of filter, cutting speed/temp, etc.) with me? Thanks so much. Angela Welford, EMT Dept of Pathology UNM-HSC Albuquerque, NM, 87131
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 13 17:18:33 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (scott.watson-at-granite.k12.ut.us) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Monday, October 13, 2003 at 17:11:52 ---------------------------------------------------------------------------
Email: scott.watson-at-granite.k12.ut.us Name: Scott Watson
Organization: Hunter High School
Education: 9-12th Grade High School
Location: West Valley City, Utah
Question: Hello. I am the electronics, engineering, and scanning electron microscopy instructor at Hunter High School in West Valley City, Utah. Here at Hunter we have an old Amray 1200C2 SEM which has been extensively modified since Amray went out of business. The microscope has had 99.9 percent of electronic circuitry replaced with new/redesigned circuitry which transformed the SEM into an ESEM. Recently several of the universal swivel joints used for adjusting the X,Y,Z, and T axises of the specimen chamber have broken. I was wondering if you knew of a source for these joints? I only need to find a source for joints - they do not have to be original manufacturer? Do you know if another SEM manufacturers specimen chambers were similar in design to the Amray 1200 and could have their specimen axis adjustment joints placed into the Amray?
Ian M. Watt, The principles and practice of Electron Microscopy, 2.ed. Cambridge University Press 1997, ISBN 0521 43591 9
very low on theory!
Philip
----- Original Message ----- } From: "Carol Heckman" {heckman-at-bgnet.bgsu.edu} To: "Microscopy Listserver" {Microscopy-at-sparc5.microscopy.com} Sent: Monday, October 13, 2003 10:46 PM
Scott, a long while back I had a conversation with a microscope designer. He mentioned that the trend is to go away from mechanical stage manipulation and use manipulation by electric motors. One conspicuous benefit was fewer mechanical intrusions into the vacuum chamber that needed sealing and maintenance. The other was overall lower cost. Should you reach a dead end with your search for a swivel joint replacement, you may want to consider an upgrade to electrical stage manipulation.
Stu Smalinskas, P.E. SKF USA Plymouth, Michigan (734) 414-6862
Scott Watson wrote:
Question: Hello. I am the electronics, engineering, and scanning electron microscopy instructor at Hunter High School in West Valley City, Utah. Here at Hunter we have an old Amray 1200C2 SEM which has been extensively modified since Amray went out of business. The microscope has had 99.9 percent of electronic circuitry replaced with new/redesigned circuitry which transformed the SEM into an ESEM. Recently several of the universal swivel joints used for adjusting the X,Y,Z, and T axises of the specimen chamber have broken. I was wondering if you knew of a source for these joints? I only need to find a source for joints - they do not have to be original manufacturer? Do you know if another SEM manufacturers specimen chambers were similar in design to the Amray 1200 and could have their specimen axis adjustment joints placed into the Amray?
Sincerely
Scott S. Watson
Email: scott.watson-at-granite.k12.ut.us Name: Scott Watson Organization: Hunter High School Education: 9-12th Grade High School Location: West Valley City, Utah
__________________________________ Do you Yahoo!? The New Yahoo! Shopping - with improved product search http://shopping.yahoo.com
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 14 08:46:10 2003
We did a survey on available textbooks about five years ago for an undergraduate course on SEM, TEM and analytical methods and we decided to use "Electron Microscopy and Analysis" by Goodhew and Humphreys. It is now published in a 3rd edition with Beanland as coauthor:
P. J. Goodhew, J. Humphreys and R. Beanland: "Electron Microscopy and Analysis" Taylor & Francis.
-----Original Message----- } From: Ian MacLaren [mailto:maclaren-at-tu-darmstadt.de] Sent: 13. oktober 2003 10:09 To: Microscopy Listserver
Dear all, What books are you using in the undergrad teaching of electron microscopy? I am in a materials department so we are most concerned with EM of inorganic materials, although polymers might be worth covering too. Also, I do mostly TEM, but I do also talk more generally about SEM and other electron beam instruments too. Does anyone have particular recommendations for textbooks for the undergraduates in this area, they will be in the fifth semester (3rd year, 1st term)?
Much as I like Williams and Carter, this is still a bit big and detailed for the target audience. Mike Loretto's book used to be quite good for this sort of thing, but times have moved on. Maybe, there are newer books on the market that cover the target audience better, with some covereage of recent developments.
Thanks in advance for any useful advice you can give.
-- Ian MacLaren Technische Universität Darmstadt Material- und Geowissenschaften Petersenstr. 23 64287 Darmstadt Germany http://www.tu-darmstadt.de/fb/ms/fg/sf/projekte/maclaren-Dateien/maclaren.html
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 14 09:23:38 2003
Postdoctoral position: Gold labels will be developed to target genetic tags on expressed proteins for identification of subunits and alignment for image processing of cryoelectron micrographs. Three areas will be coordinated: 1) Chemical design and synthesis of specialized gold nanoparticles, 2) genetic expression of tagged proteins, 3) cryoEM using JEOL 2010F 200keV field emission with 2k CCD and STEM attachment. Position will involve all areas, but the major emphasis will be in using the EM and performing reconstructions. Contact Dr. James Hainfeld, Brookhaven National Lab, Biology Dept., Long Island, NY, hainfeld-at-bnl.gov.
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 14 15:06:27 2003
Dear All: When I use Lowicryl HM20, it sometimes turns to light pink color after polymerization. Has anyone else experienced this? Does anyone know why it changes color? Thank you much?
Hong
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 14 16:07:59 2003
I have recently collected some large (13.8M) 16 bit grayscale tiff images from my SEM. From the SEM software they look very good and print well to my inkjet printer. The problem I am having is reading the files into any 3rd party imaging software (ie Photoshop, Photo Impact) for resizing and saving in additional formats such as jpeg. Is there software out there that can handle such files or am I missing something about the tiff file format?
Thanks
Roy Beavers Southern Methodist University Department of Geological Sciences P.O. Box 750395 Dallas, Tx. 75275 Voice: 214-768-2756 Fax: 214-768-2701 Email: rbeavers-at-mail.smu.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 14 17:43:01 2003
Have you tried the good old IrfanView (free from www.irfanview.com)?
cheers
rtch
Is there anybody out there successfully sectioning large Immunobed or JB-4 blocks with Ralph knives or Tungsten Carbide knives? What is the practical limit to the specimen width?
Ralph Common Electron Microscopist Michigan State University Division of Human Pathology A608 East Fee Hall East Lansing, MI 48824 517-355-7558; fax 517-432-1053 ralph.common-at-ht.msu.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 14 19:33:34 2003
I've occasionally encountered problems opening tif files in Photoshop, perhaps because of header info that did not conform in some way to what was expected. In those cases I was able to open them in UTHSCSA Image Tool (U. of Texas) or Irfanview and immediately re-save them from those applications. Once saved from one of those applications I could open them in Photoshop without further problems. I never pursued the issue further.
John Twilley
Beavers, Roy 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 } ------------------------------------------------------------------------------- } } Group, } } I have recently collected some large (13.8M) 16 bit grayscale tiff images from my SEM. From the SEM software they look very good and print well to my inkjet printer. The problem I am having is reading the files into any 3rd party imaging software (ie Photoshop, Photo Impact) for resizing and saving in additional formats such as jpeg. Is there software out there that can handle such files or am I missing something about the tiff file format? } } Thanks } } Roy Beavers } Southern Methodist University } Department of Geological Sciences } P.O. Box 750395 } Dallas, Tx. 75275 } Voice: 214-768-2756 } Fax: 214-768-2701 } Email: rbeavers-at-mail.smu.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 14 19:42:40 2003
ImageJ, the free program from NIH, should have no trouble reading the file, but it depends on the available memory in your system. You can get ImageJ here: http://rsb.info.nih.gov/ij/
When you install it, read the section on memory management. You will have to tweak the command line in the shortcut to make all of your machine's memory available.
Good luck
Joel
Hi Chris I think you are right. When I immersed half film in the fixer - after a few hours I did see noticeable bleached line on the border between fixer and air. This line is about 1 mm thick by the way. It clearly indicates for me that air is involved (most noticeable on the water-air interface). Except that interface line, the effect of bleaching was about 3% from untouched film for 1.5 h exposure to the fixer. My guess is that at fixer-air interface the metal Ag is more available for the oxidation and then bleaching by the sodium thiosulphate as it correctly cited in your last posting. 3% bleaching effect inside the solution is not big deal: variation in temperature/time perhaps will benefit more pronounced effect on the film. But still, I am impressed: this is a pure example how real life interfere with our basic knowledge without any respect to the "theory"... Anyway, thanks for pointing out this problem. Sergey.
At 02:16 AM 10/13/2003, you wrote:
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_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
14MB is not all that large of a TIFF file. If it is 16-bits, then it is actually 7MB as 8-bit.
Load the file into PS and do a Image/Mode change from 16-bit to 8-bit and you can do most anything you want to. Make sure it is not indexed RGB but pure and simple grey scale.
gary g.
At 02:09 PM 10/14/2003, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Tue Oct 14 23:01:55 2003
The price is right but the performance is not. I get too many "out of memory" errors.
I suppose that 2GB RAM is not enough. IJ is OK for small files but not for big or larger ones, IMOH.
gary g.
At 05:47 PM 10/14/2003, you wrote:
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From MicroscopyL-request-at-ns.microscopy.com Wed Oct 15 03:05:56 2003
I am trying to prepare some samples for TEM analysis, lattice-fringes images. I have different suspensions which include Wyoming montmorillonite and iron oxides. I tried to obtain oriented aggregates by filtration, but I did not managed. I also tried the sedimentation way, but it needs time and at the end, the mineral phases are not the same (the iron oxides may transform in time and may also interact with the clay mineral). I am trying now to use the supercentrifuge, but I do not know if I will manage to take the oriented aggregate without disturbing them. I should need an advice. I should need at the end to use the L.R.White resin to keep the montmorillonite interlayer hydrated. Thank you
----------------------------------------------------------------------------------- Prof. Elena BELLUSO - Ph.D. Mineralogy and Crystallography Dipartimento di Scienze Mineralogiche e Petrologiche Università degli Studi di Torino Via Valperga Caluso, 35 I-10125 TORINO - ITALIA tel: (39) 011 670 71 35 - fax: (39) 011 670 71 28 e-mail: elena.belluso-at-unito.it {http://www.dsmp.unito.it/} http://www.dsmp.unito.it ----------------------------------------------------------------------------------- "I've... seen things you people wouldn't believe. Attack ships on fire off the shoulder of Orion. I watched C-beams... glitter in the dark near the Tanhauser Gate. All those... moments will be lost... in time..., like... tears... in... rain." Blade Runner
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From MicroscopyL-request-at-ns.microscopy.com Wed Oct 15 05:43:33 2003
Hello Hong, For HM20 it is just the normal color whe block is polymerized. The K4M is jelowish and the HM20 reddish when exposed to air Best regards Danièle
-----Message d'origine----- De : Hong Yi [mailto:hyi-at-emory.edu] Envoyé : mardi 14 octobre 2003 22:07 À : Microscopy-at-msa.microscopy.com Objet : Lowicryl HM20
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Dear All: When I use Lowicryl HM20, it sometimes turns to light pink color after polymerization. Has anyone else experienced this? Does anyone know why it changes color? Thank you much?
Hong
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 15 06:37:02 2003
Is it possible to explain how you would like to keep the hydraded stucture in TEM? From my experience the resin embedding will modify the structure. Unless you need to do cryo-sectioning and cryo-TEM.
My suggestion is that if your suspension is not very dilute, you could try a direct collection on a Cu-grid with a carbon film. For TEM application this would be perfectly sufficient and you will not induce any interaction of the two constituents. After collection of the sample, if you obtain a very dense distribution you can embed directly and immediatedly with the grid.
The other alternative is to do cryo-TEM using the suspension directly on the grid followed by rapid cooling and cryo-TEM. For that you need specialised laboratories and if you were interested I could inform you.
Hope this helps, Sousan
Dr. Sousan Abolhassani Laboratory for Materials Behaviour Paul Scherrer Institut 5232 Villigen PSI Switzerland
Elena Belluso 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 am trying to prepare some samples for TEM analysis, lattice-fringes } images. I have different suspensions which include Wyoming montmorillonite } and iron oxides. } I tried to obtain oriented aggregates by filtration, but I did not managed. } I also tried the sedimentation way, but it needs time and at the end, the } mineral phases are not the same (the iron oxides may transform in time and } may also interact with the clay mineral). } I am trying now to use the supercentrifuge, but I do not know if I will } manage to take the oriented aggregate without disturbing them. } I should need an advice. I should need at the end to use the L.R.White } resin to keep the montmorillonite interlayer hydrated. } Thank you } } ----------------------------------------------------------------------------------- } Prof. Elena BELLUSO - Ph.D. Mineralogy and Crystallography } Dipartimento di Scienze Mineralogiche e Petrologiche } Università degli Studi di Torino } Via Valperga Caluso, 35 } I-10125 TORINO - ITALIA } tel: (39) 011 670 71 35 - fax: (39) 011 670 71 28 } e-mail: elena.belluso-at-unito.it } {http://www.dsmp.unito.it/} http://www.dsmp.unito.it } ----------------------------------------------------------------------------------- } "I've... seen things you people wouldn't believe. } Attack ships on fire off the shoulder of Orion. } I watched C-beams... glitter in the dark near the Tanhauser Gate. } All those... moments will be lost... in time..., } like... tears... in... rain." } Blade Runner } } _____________________________________________________________________ } For your security, this mail has been scanned and protected by Inflex
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 15 06:47:05 2003
Laboratory Manager position opening in NYC in multi-user microscopy lab.
Responsibilities include assisting and training staff in scanning electron microscopy (SEM), x-ray microanalysis (EDS), confocal laser scanning microscopy (CLSM), and cathodoluminescence spectroscopy (CLS). Additional responsiblities include assisting and training staff with image processing, and printing for publication and presentation. General lab duties include PC/MAC hardware and software maintenance/upgrades, ordering consumables, and scheduling equipment maintenance.
The ideal candidate will have a B.S. in a natural science, 2-3 yrs experience with SEM theory and operation, and experience with computer hardware/software. Good interpersonal skills are essential.
Contact Dr. Angela V. Klaus via email at avklaus-at-amnh.org. EOE.
----------------------------------------- Angela V. Klaus, Ph.D. Director, Microscopy and Imaging Facility American Museum of Natural History Central Park West at 79th Street New York, NY 10024 USA Email: avklaus-at-amnh.org Tel: 212-769-5977 Fax: 212-496-3480 -----------------------------------------
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 15 07:04:03 2003
} I have recently collected some large (13.8M) 16 bit } grayscale tiff images from my SEM. From the SEM software } they look very good and print well to my inkjet printer. } The problem I am having is reading the files into any 3rd } party imaging software (ie Photoshop, Photo Impact) for resizing } and saving in additional formats such as jpeg. } Is there software out there that can handle such files or am } I missing something about the tiff file format?
I am surprised Photoshop couldn't open them! ... although there is always that possibility, but I also imagine other softwares would have a similar problem with an inaccurate TIFF format. If you know the size of the image, then try opening them "as raw", and then filling in the appropriate info ... e.g., 1024 pixels horizontal, 807 pixels vertical, 16bits deep, and have PS guess at the header size.
If the header size is wrong, then you'll see an image with a border down the middle, and the left & right parts of the image opposite. You can then reopen and guess at the header again, and the correct guess should be accurate for all other images. There is a more systematic approach to guessing the header size, and you can e-mail me as I need to find it ... but the clue is in the row of pixels at the top of the image.
I'll admit that I'm frustrated with my TEM vendor (out of MA). I have an "older" 200KV TEM, but have faithfully kept it under service contract for over 10 years. Given its age it is experiencing some failures, as one would expect. My frustration is that the vendor seems to be trying to fix the same problem over and over without coming to a conclusion. This is causing a huge amount of downtime (since the summer i've not had more than a week of contiguous uptime). When it fails usually it takes 3-4 days or a week to get someone on-site, although they often give me some things to check or "try". The net result is that my time is chewed up and still I have no operating TEM. To be honest, my financial situation requires that I purchase a time-and-parts limited contract at discount, but it seems that the current situation would be unacceptable even with a full contract. With the current uptime percentage I can't even break even on reimbursement for the limited contract from my userbase. Using automobile service as a comparison, I'd be very upset if it was in the shop or awaiting service (for the same or related failures) as much as this TEM is. My question to the group is: what can reasonably be expected for a generation-or-two old TEM for uptime, and how can i work with the vendor to maximize the impact of service calls? Thanks! Brian
_________________________________________________ Brian McIntyre Univ. of Rochester The Institute of Optics River Campus EMLab 585-275-3058/4875 585-244-4936 (fax)
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 15 10:31:13 2003
Thank you for your answer. I do not know if I can obtain my sample using your first proposed method. I am trying to explain you better what I need. I must prepare some systems composed by iron oxides and Wy-montmorillonite, using different ratios iron oxide/montmorillonite in systems with 10 g solid/1 l suspension. I intend to study the mineral phases modifications in time. By TEM, I hope to observe the behavior of d(001) in different moments of the reaction. My intention was to separate the solid, obtaining first an oriented aggregate. After I would want to embed this aggregate with L.R. White resin, following the procedure described by Kim J.-W., Peacor D.R., Tessier D., Elsass F. (1995) A technique for maintaining texture and permanent expansion of smectite interlayer for TEM observations, Clays Clay Miner., 43, 51-57. This procedure allows to preserve the d spacing of montmorillonite unmodified. Thus, after the embedding, I would use a microtome to obtain the TEM samples from slides perpendicular on the 001 planes. Do you think that using your first method I could arrive at this? I already tried to use the sedimentation on glass. I managed obtain a small disc, but very, very thin in time. I put some drops, after I waited to dry, I put another drops etc. I tried also to put directly a greater quantity, limiting my disc with a vertical tube, but an important quantity of solid (the finest) remained on the walls. Anyway, to obtain a disc of 3 mm thickness I would need much time and the initial suspension will not be the same at the end. I need to separate the solid quickly to preserve this composition. I am trying now the supercentrifuge method. I hope to recover the solid, cutting the plastic tube. I read about the rapid cooling, but using these technique I will have crystals randomly oriented. I hope that now I managed to explain better my problem.
Thank you again
----------------------------------------------------------------------------------- Prof. Elena BELLUSO - Ph.D. Mineralogy and Crystallography Dipartimento di Scienze Mineralogiche e Petrologiche Università degli Studi di Torino Via Valperga Caluso, 35 I-10125 TORINO - ITALIA tel: (39) 011 670 71 35 - fax: (39) 011 670 71 28 e-mail: elena.belluso-at-unito.it {http://www.dsmp.unito.it/} http://www.dsmp.unito.it ----------------------------------------------------------------------------------- "I've... seen things you people wouldn't believe. Attack ships on fire off the shoulder of Orion. I watched C-beams... glitter in the dark near the Tanhauser Gate. All those... moments will be lost... in time..., like... tears... in... rain." Blade Runner
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From MicroscopyL-request-at-ns.microscopy.com Wed Oct 15 10:33:24 2003
I got pink color with HM 20 after polymeration too. The color fades in a few days. I asked around about the reason and didn't get the answer yet.
Shanling
-----Message d'origine----- De : Hong Yi [mailto:hyi-at-emory.edu] Envoyé : mardi 14 octobre 2003 22:07 À : Microscopy-at-msa.microscopy.com Objet : Lowicryl HM20
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Dear All: When I use Lowicryl HM20, it sometimes turns to light pink color after polymerization. Has anyone else experienced this? Does anyone know why it changes color? Thank you much?
Hong
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 15 10:53:20 2003
If you're using a Mac, there's an excellent shareware programme called Graphic Converter, which can read anything and translate it into anything else.
http://www.lemkesoft.com/
Hope this helps.
Lesley Weston.
on 14/10/2003 3:44 PM, Ritchie Sims at r.sims-at-auckland.ac.nz 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 } ---------------------------------------------------------------------------- --} - } } } Have you tried the good old IrfanView (free from www.irfanview.com)? } } cheers } } rtch } } } Subject: [Microscopy] Image reader } Date sent: Tue, 14 Oct 2003 16:09:02 -0500 } } From: "Beavers, Roy" {rbeavers-at-mail.smu.edu} } To: "Microscopy Listserver" {Microscopy-at-sparc5.microscopy.com} } } } } } } } ---------------------------------------------------------------------- } } -------- 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 } } ---------------------------------------------------------------------- } } --------- } } } } Group, } } } } } } I have recently collected some large (13.8M) 16 bit grayscale tiff } } images from my SEM. From the SEM software they look very good and } } print well to my inkjet printer. The problem I am having is reading } } the files into any 3rd party imaging software (ie Photoshop, Photo } } Impact) for resizing and saving in additional formats such as jpeg. } } Is there software out there that can handle such files or am I } } missing something about the tiff file format? } } } } Thanks } } } } Roy Beavers } } Southern Methodist University } } Department of Geological Sciences } } P.O. Box 750395 } } Dallas, Tx. 75275 } } Voice: 214-768-2756 } } Fax: 214-768-2701 } } Email: rbeavers-at-mail.smu.edu } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 15 11:19:49 2003
I'm looking for a vacuum gauge power supply for an Hitachi HU125E similar to the HU11E series built in the late 60s early 70s (you remember, back when music was good). The supply was typically housed in a rack along with the beam deflector/stigmator module.
I'm also interested in other parts from the HU125E series, all going to support undergraduate EM training.
Contact me off list if you can lend a hand.
Thanks!
Robert -- Robert Fitton Teaching Associate/Director of Laboratories Luther College Department of Biology 700 College Drive Decorah, IA 52101 fittonro-at-luther.edu Voice 563-387-1559 FAX 563-387-1080
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 15 13:07:25 2003
For grayscale image, most standard image format can support only 256 greyness scale(that means the image depth can only reach 8 bits). Tiff is a flexible format and it can be extended very easily. Many people just extend it to 16 bits or 32 bits. However, this extension can not be recognized by most image processing software.
I think your SEM manufactuer should provide you some ways to change it to other file formats or standard tiff format. If they didn't do it, they were very irresponsible.
If there were no other ways to do it. I suggest you write some programs yourself. You don't need to start from scratch. There are many tiff libraries which can be downloaded from web. Besides, in the book "Advanced computation in Electron Microscopy", Earl Kirkland gave some source code to cope with 32 bits extended tiff file format. I guess you can just revise it to cope with 16 bits.
By the way, opening tiff file is not related to the memory of your computer, even though some software told you that your system lacks memory.
} } } ------------------------------------------------------------------------------ } 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 } ------------------------------------------------------------------------------ - } } Group, } } } I have recently collected some large (13.8M) 16 bit grayscale tiff images } from my SEM. From the SEM software they look very good and print well to my } inkjet printer. The problem I am having is reading the files into any 3rd } party imaging software (ie Photoshop, Photo Impact) for resizing and saving } in additional formats such as jpeg. Is there software out there that can } handle such files or am I missing something about the tiff file format? } } Thanks } } Roy Beavers } Southern Methodist University } Department of Geological Sciences } P.O. Box 750395 } Dallas, Tx. 75275 } Voice: 214-768-2756 } Fax: 214-768-2701 } Email: rbeavers-at-mail.smu.edu } } }
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From MicroscopyL-request-at-ns.microscopy.com Wed Oct 15 13:36:43 2003
Most imaging applications do not support 16-bit TIFF files, including Adobe Photoshop (which is strange, because Adobe maintains the TIFF standard). You could use our software, Image-Pro Plus version 5.0. It supports 16- and 48-bit TIFF. You can convert them to 12-, 8-, 24-, 36-bit images as well as JPEG, TGA (Targa), BMP, PICT, CUT (the Media Cybernetics standard from 20 years ago- still popular), PCX, EPS, flat file, and AVI (for multi-TIFF files).
Best Regards,
Dean Sequera V.P. Marketing & Product Development Media Cybernetics, Inc. 8484 Georgia Ave. Suite 200 Silver Spring, MD 20910 (301) 495-3305 x 262 (301) 495-5964 FAX http://www.mediacy.com
} Subject: [Microscopy] Image reader } Date sent: Tue, 14 Oct 2003 16:09:02 -0500 } From: "Beavers, Roy" {rbeavers-at-mail.smu.edu} } To: "Microscopy Listserver" {Microscopy-at-sparc5.microscopy.com} } } } } ---------------------------------------------------------------------- } -------- 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 } ---------------------------------------------------------------------- } --------- } } Group, } } } I have recently collected some large (13.8M) 16 bit grayscale tiff } images from my SEM. From the SEM software they look very good and } print well to my inkjet printer. The problem I am having is reading } the files into any 3rd party imaging software (ie Photoshop, Photo } Impact) for resizing and saving in additional formats such as jpeg. } Is there software out there that can handle such files or am I } missing something about the tiff file format? } } Thanks } } Roy Beavers } Southern Methodist University } Department of Geological Sciences } P.O. Box 750395 } Dallas, Tx. 75275 } Voice: 214-768-2756 } Fax: 214-768-2701 } Email: rbeavers-at-mail.smu.edu
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 15 14:47:39 2003
Paul--I am using a Falcon cell culture insert with an 0.4micron pore size. It incorporates polyethylene terephthalate (PET) into the membrane. Can you recommend a more preferable one from your experience?
Thanks, Angela Welford
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 15 15:13:05 2003
I would love to locate an additional wehnelt cylinder for our JSM-840. Please keep me in mind if you have an additional one you could part with or are planning to mothball or replace this model SEM.
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
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 15 18:06:57 2003
Hi Chris I think, you guys have some special oxygen, more aggressive, I guess. So, your butter should be softer and brain healthier. You right, it's 7 levels of grey. I don't understand what you meant talking about accuracy? Of coarse I did average for the area about 2cm^2 in both control and treated with fixer. But still, I did not expect to have even such effect. Thanks for the good point. Sergey.
At 12:10 AM 10/15/2003, you wrote: } Sergey } I estimated the effect as a bit stronger than that, but maybe the } oxygen concentration is greater in this part of the world. That could } account for all sorts of stuff - the sheer quality of Scottish } science, the frequency of Glasgow house fires... } To be serious for a moment, 3-5% reduction may not always be important } pictorially if it takes place uniformly, but that is unlikely to } occur in a neglected fixer bath. Also converting the effect of 3-5% } reduction into grey levels you might expect a density change of } about -7 to -13 which won't contribute any accuracy to your } densitometry. } Chris } } Dr. Chris Jeffree } Edinburgh University } Biological Sciences EM Facility } } ----- Original Message ----- } From: "Sergey Ryazantsev" {sryazant-at-ucla.edu} } To: {Microscopy-at-sparc5.microscopy.com} } Sent: Wednesday, October 15, 2003 3:29 AM } Subject: [Microscopy] Re: Re: Fixer life SO163 Development } } } } } } } } -------------------------------------------------------------------- } ---------- } } 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 Chris } } I think you are right. When I immersed half film in the fixer - } after a } } few hours I did see noticeable bleached line on the border between } fixer } } and air. This line is about 1 mm thick by the way. It clearly } indicates } } for me that air is involved (most noticeable on the water-air } } interface). Except that interface line, the effect of bleaching was } about } } 3% from untouched film for 1.5 h exposure to the fixer. My guess is } that } } at fixer-air interface the metal Ag is more available for the } oxidation } } and then bleaching by the sodium thiosulphate as it correctly cited } in your } } last posting. 3% bleaching effect inside the solution is not big } deal: } } variation in temperature/time perhaps will benefit more pronounced } effect } } on the film. But still, I am impressed: this is a pure example how } real } } life interfere with our basic knowledge without any respect to the } } "theory"... Anyway, thanks for pointing out this problem. Sergey. } } } } At 02:16 AM 10/13/2003, 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 } } } } --------------------------------------------------------------------- } ---------- } } } } } } Sergey } } } I don't pretend to be able to account for this effect, but it } happens } } } anyway. Try } } } this simple test: stand an unwanted negative on edge with lower } half immersed } } } in a bath of your favourite acid or rapid fixer overnight. Wash and } dry. } } } Compare } } } density of immersed and non-immersed areas. } } } } } } See quotes below from } } } Grant Haist (1979) Modern Photographic Processing; John Wiley & } Sons, } } } Inc.; New York; Vol. 1; pps. 564-565 } } } "Another danger of long fixing bath immersions is the direct attack } on the } } } silver image by the combination of oxygen and the acid fixing bath. } It is } } } believed that oxygen from the air dissolves in the fixing bath and } attacks } } } the } } } very finely divided silver particles of the image. Oxygen converts } these } } } metallic silver particles to silver ions by removing electrons. The } silver } } } ions } } } are then complexed by the thiosulfate and removed, resulting in the } loss of } } } image silver. Fine-grained films and paper images are especially } susceptible } } } to image reduction by acid thiosulfate solutions." } } } "The extent of the reducing effect of fixing baths on the silver } image during } } } the process of fixation is greater than has been generally } supposed. This } } } attack does not occur in alkaline thiosulfate solutions" } } } Best wishes } } } Chris } } } } } } } } } On 10 Oct 03, at 20:38, 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 } } } } } -------------------------------------------------------------------- } -- } } } } --------- } } } } } } } } Chris } } } } Thanks for your message. It forced me to refresh my memory in } } } } chemistry. The process of dissolving of metal silver is } reduction. } } } } Metal Ag in the film is already reduced in compare with silver } halide. } } } } The conditions in the fixer are again reducing, so there is no } } } } chemical direct way to reduce metal silver in the fixer. It } meant that } } } } directly sodium thiosulphate could not dissolve the metal silver } in } } } } the film but silver halide. There are couple of things you need } to } } } } keep in mind: if for some reason silver is oxidized, then you } could } } } } reduce it with sodium thiosulphate and therefore "dissolve it". } } } } Because general reducing conditions in the fixer, oxygen from } air may } } } } not be effective. You need to add something like bleach into } the } } } } fixer to oxidize silver and then you may reduce it with sodium } } } } thiosulphate. Another thing comes to my mind is chlor gas. So, } you } } } } may keep film in the fixer for quite while and silver will not } } } } dissolved (if only you will not add the bleach). Professional } } } } photographers usually do not recommend to keep film in fixer for } so } } } } long for another reason: sodium thiosulphate is slowly } decomposing } } } } with creation of elementary sulphur, which colored film in } yellowish } } } } color. If you will keep your film in the fixer for couple of } days, } } } } elementary sulfur will be deposited in the gelatin layer, which } made } } } } film yellowish irreversibly. Similar thing is happening when you } do } } } } not wash film well after fixer - the residue of sodium } thiosulphate } } } } will decompose with sulphur creation. So, it's good practice to } ensure } } } } your film is washed well after fixer. Have a great wekend, } Sergey } } } } } } } } At 03:08 AM 10/10/2003, 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 } } } } } } --------------------------------------------------------------------- } } } } } ---------- } } } } } } } } } } Sergey } } } } } Sodium thiosulphate has a mild solvent effect on silver, and } this is } } } } } accelerated at low pH in acid and hardening fixer formulations. } The } } } } } consequences are negligible during standard fixing times for } film or } } } } } paper, but prolonged over-fixing will produce noticeable } bleaching, } } } } } reducing overall negative or print density and bleaching out } low } } } } } density areas so that shadow detail in negatives and highlight } detail } } } } } in prints are lost. } } } } } } } } } } An old rule of thumb to determine the required fix time for EM } film } } } } } in old or partially-used fixer is twice the time taken to clear } the } } } } } film. Another rule of thumb is to discard the fixer when the } clearing } } } } } time is twice that for a fresh solution. Development is stopped } very } } } } } quickly in a fix bath unless it is completely exhausted, and it } is } } } } } perfectly safe to inspect the negatives as they clear in } safelight or } } } } } even in subdued white light. The most serious downside of } stretching } } } } } fixer life to the limit is that it increases the risk of silver } } } } } staining of the negatives, and may reduce their archival } permanence. } } } } } } } } } } I gave a formula for D19 on this list some months back. Here it } is } } } } } again, derived from British Journal of Photography almanac. } } } } } } } } } } Metol 2.2g, Hydroquinone 8.8g, Sodium sulphite 144g, Sodium } carbonate } } } } } 130g Potassium bromide 4g, H2O to 1litre } } } } } } } } } } However, other sources quote markedly different proportions of } the } } } } } ingredients, e.g. this one from Leica Users group } } } } } http://mejac.palo-alto.ca.us/leica-users/v03/msg13172.html D19: } water } } } } } 500 cc Metol 2.0 grams sodium sulfite, desc. 90.0 grams } Hydroquinone } } } } } 8.0 grams sodium carbonate, monohyd. 52.5 grams potassium } bromide 5 } } } } } grams cold water to make 1.0 liter and this one from } } } } } http://astro.umsystem.edu/apml/ARCHIVES/MAR01/msg01118.html You } can } } } } } make D19 yourself by using its formula: Metol 2 g } } Sodium } Sulfite } } } } } 90 g } } Hydroquinone 8 g } } Sodium Carbonate (anhydrous) 45 g } } } } } } } Potassium Bromide 5 g } } Water 1 liter } } } } } } } } } } What is the official formula of Kodak D19 as currently } marketed. } } } } } Anyone have authoritative information? } } } } } } } } } } } } } } } Dr. Chris Jeffree } } } } } University of Edinburgh } } } } } Biological Sciences EM Faciility } } } } } } } } } } ----- Original Message ----- } } } } } } From: "Sergey Ryazantsev" {sryazant-at-ucla.edu} } } } } } To: {Microscopy-at-sparc5.microscopy.com} } } } } } Sent: Friday, October 10, 2003 4:49 AM } } } } } Subject: [Microscopy] Re: SO163 Development } } } } } } } } } } } } } } } } } } } } } } } } } } } } } ------------------------------------------------------------------ } } } } } } -- } } } } } ---------- } } } } } } 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 } } } } } } } ------------------------------------------------------------------ } } } } } } -- } } } } } ----------- } } } } } } } } } } } } Garry } } } } } } You are right and aren't. Kodak spent a lot of time (and } perhaps } } } } } money) to } } } } } } develop chemical formulations, which are stable over some } period } } } } } } of } } } } } time } } } } } } being in use. It, actually, made a revolution in the } photographic } } } } } process: } } } } } } all C-41 process machines in our drug-stores used the } solutions, } } } } } which } } } } } } stable for quite a while. It dramatically reduces the cost } of } } } } } developing } } } } } } and maintenance. Similarly they invented "long-play" } solutions to } } } } } develop } } } } } } X-rays films in the hospitals (we do have such machine - } } } } } } technician } } } } } changed } } } } } } solutions ones a month). Because in TEM it's just } impossible to } } } } } } use } } } } } fresh } } } } } } developer every time you need to develop a few films and } Kodak } } } } } suggest to } } } } } } use "used" developer over some period of time, I suspect } D-19 has } } } } } "extended } } } } } } life" formulation as well. Based on my personal experience, } it's } } } } } clear to } } } } } } me that diluted solution of D-19 (used or non-used) deliver } about } } } } } the same } } } } } } results for at least 3 weeks if properly stored without } exposure } } } } } } to air. Usually we develop about 100 films in 1.5 liter. I } } } } } } think, the } } } } } decent } } } } } } quality of the TEM film developed in used developer is also } } } } } } because } } } } } of } } } } } } special film formulation (bye-bye old 4489!). It's called } } } } } } "electron microscopy film" and I assume it separates this } film } } } } } } from other type } } } } } of the } } } } } } films. As for price for chemicals - please count your } technician } } } } } time to } } } } } } make fresh solutions 3 times a day and utilize used ones, } it's not } } } } } cheap as } } } } } } you think. On the positive side, I do agree with you that } it's } } } } } good idea } } } } } } in general to use fresh components everywhere: in } photography, } } } } } } biochemistry, cooking etc. Sergey } } } } } } } } } } } } P.S. You don't need to change Stop-bath and fixer so often. } I do } } } } } believe, } } } } } } that Kodak suggests that stop-bath solution is good for } 1000+ } } } } } films... The } } } } } } things about fixer - it could not spoil your film as long as } you } } } } } careful do } } } } } } not mix it with developer. You may left film in the fixer } for } } } } } } hours } } } } } and it } } } } } } does not harm film. You need to wash it after all by the } way. } } } } } } The } } } } } bottom } } } } } } line here is that you may extend fixation time if your } fixer } } } } } } start } } } } } working } } } } } } slowly saving money and doing good for environment (less } chemical } } } } } waste). } } } } } } } } } } } } } } } } } } At 07:46 PM 10/9/2003, you wrote: } } } } } } } While I don't use SO163 film or compatible developers, } } } } } } } I disagree in general about the life of film developer mix. } } } } } } } The film developer is the most important step in } } } } } } } processing silver halide media. At least, this is } } } } } } } how I see it. } } } } } } } } } } } } } } I use (or would have used) fresh developer for each } } } } } } } batch of film that I developed. And Kodak explicitly } } } } } } } details (as do others) how many rolls or sheets of film } } } } } } } can be processed with a fresh batch before refreshing } } } } } } } or dumping a pot of developer. I never refreshed } } } } } } } developer. The results are just not the same as with } } } } } } } a fresh batch. } } } } } } } } } } } } } } It may be that this TEM film is pathetically ignorant } } } } } } } of basic photo chemistry. However, for such a small } } } } } } } cost of chemicals, why take the chance? If the shots } } } } } } } are worth shooting, they are worth proper processing. } } } } } } } That means new chemicals. And, these chemicals must } } } } } } } be at the correct temperature to be consistent and } } } } } } } effective. I've seen too many instances of no temperature } } } } } } } control. Consequently, the results vary wildly. Duh. } } } } } } } } } } } } } } Furthermore, I dumped stop bath and fixer routinely. } } } } } } } Why cut corners after so much work on the instrument? } } } } } } } } } } } } } } gary g. } } } } } } } } } } } } } } } } } } } } } } } } } } } } At 05:12 PM 10/9/2003, 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 } } } } } } } } } } } } } } ------------------------------------------------------------------ } } } } } } } -- } } } } } ----------- } } } } } } } } } } } } } } } } Basically, the life of unused (even diluted) developer is } quite } } } } } long } } } } } } } } (without air). The clock starts ticking as soon as you } develop } } } } } even one } } } } } } } } piece of the film (similar but less dramatic for paper } } } } } developers). The } } } } } } } } amount of film company recommends to develop usually mean } that } } } } } } } } you supposed to develop all those films in relatively } short } } } } } } } } period of time. Classical example is quite popular in } amateur } } } } } } } } photography } } } } } D-76 } } } } } } } } developer. In 1 liter of the fresh developer you could } develop } } } } } about 5 } } } } } } } } rolls of 35 mm film, but developer will die in about 4-5 } hours } } } } } does not } } } } } } } } matter how many films you manage to develop, so hurry up! } } } } } Similarly, } } } } } } } } it's true for D-19 especially non-diluted. It will not } die in } } } } } } } } few } } } } } hours, } } } } } } } } but perhaps two weeks. The exposure to air is very } critical } } } } } } } } here also. So, the bottom line here is: if you need to } develop } } } } } } } } a lot } } } } } of films } } } } } } } } in quite short period of time - you may do it to extend } } } } } } } } company's recommendation. As soon as you start use } developer, it } } } } } } } } will } } } } } gradually } } } } } } } } lost capacity in the matter of few weeks and will die even } if } } } } } } } } you } } } } } just } } } } } } } } developed 10 films. Note: this does not applied to the } fixer - } } } } } } } } as } } } } } long } } } } } } } } as you do not contaminate it with developer, it will works } to } } } } } } } } the } } } } } end of } } } } } } } } capacity, does not matter how long it will take (6 mo for } sure } } } } } } } } and } } } } } even } } } } } } } } longer). Fixer will die soon if you contaminate it with } } } } } developer. By } } } } } } } } the way - if you are using those test drops to check } fixer - } } } } } } } } they } } } } } are too } } } } } } } } sensitive, you may use fixer much longer and for more } films } } } } } without any } } } } } } } } harm to the film. Experimentally I determined that when } those } } } } } "drops" } } } } } } } } indicate, the fixer is bad, it actually used about 50% of } } } } } capacity. I } } } } } } } } hope it helps. Sergey } } } } } } } } } } } } } } } } } } } } } } } } At 06:11 AM 10/9/2003, 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 Leslie: } } } } } } } } } } } } } } } } } } 12 minutes in straight D-19 sounds like a huge push } to me! } } } } } } } } } On } } } } } the } } } } } } } } } other hand, I have no experience with cryo EM so ... } Have you } } } } } talked to } } } } } } } } } Kodak about this? Long times in developer can lead to } fogging. } } } } } } } } } As to life of the chemistry in a tank. If you have a } } } } } } } } } floating } } } } } lid } } } } } } } } } (that excludes most air) the chemistry will keep better } than } } } } } } } } } if } } } } } the } } } } } } } } } lid allows free access to air. Again, a call to Kodak } is in } } } } } order. As } } } } } } } } } for the number of films I always stay on the } conservative } } } } } } } } } side, } } } } } maybe } } } } } } } } } 2/3 of the recommendation. } } } } } } } } } D-19 is the cheapest part of the equation (animals, } prep } } } } } time, etc) } } } } } } } } } so trying to squeez a few extra films out of a pot of } } } } } } } } } developer } } } } } is not } } } } } } } } } the way to go. Also, D-19 does not last more than 2 } years in } } } } } } } } } the package on the shelf. I don't do much TEM anymore so } I mix } } } } } } } } } my } } } } } D-19 from } } } } } } } } } scratch using the individual chemicals, not the } prepackaged } } } } } Kodak mix. } } } } } } } } } The recipe is widely published, Kodak will give it to } you if } } } } } } } } } you } } } } } ask. } } } } } } } } } } } } } } } } } } Geoff } } } } } } } } } } } } } } } } } } Leslie Cummins wrote: } } } } } } } } } } } } } } } } } } } Hello Listers } } } } } } } } } } } } } } } } } } } } We are setting up to develop Kodak SO163 film for low } dose } } } } } cryoEM. We } } } } } } } } } } are push processing it, so we are developing it for 12 } minutes } } } } } in } } } } } } } } } } straight D-19, we're using a nitrogen gas burst, for 1 } second } } } } } every 8 seconds. } } } } } } } } } } } } } } } } } } } } I was wondering what the life of the chemistry might be. } } } } } } } } } } Kodak } } } } } said } } } } } } } } } } that we could develop 60 8x10 negatives per gallon. } This } } } } } } } } } } works } } } } } out to } } } } } } } } } } about 360 3 1/4 x 4 negatives. This seems like a lot of } } } } } negatives to } } } } } } } } } } us, so we are wondering what other people are using for } an } } } } } exhaustion } } } } } } } } } } time, both in negative numbers and days in the tank. } } } } } } } } } } } } } } } } } } } } Thanks for the help. } } } } } } } } } } Leslie Cummins } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } Leslie Gunther Cummins } } } } } } } } } } Analytical Imaging Facility } } } } } } } } } } Albert Einstein College of Medicine } } } } } } } } } } 1300 Morris Park Ave. } } } } } } } } } } Bronx, NY 10461 } } } } } } } } } } 718-430-3547 } } } } } } } } } } } } } } } } } } } } http://www.aecom.yu.edu/aif/ } } } } } } } } } } } } } } } } } } -- } } } } } } } } } -- } } } } } } } } } ********************************************** } } } } } } } } } 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 } } } } } } } } } ********************************************** } } } } } } } } } } } } } } } } } } } } } } } } } _____________________________________ } } } } } } } } } } } } } } } } Sergey Ryazantsev Ph. D. } } } } } } } } Electron Microscopy } } } } } } } } UCLA School of Medicine } } } } } } } } Department of Biological Chemistry } } } } } } } } 10833 Le Conte Ave, Room 33-089 } } } } } } } } Los Angeles, CA 90095 } } } } } } } } } } } } } } } } Phone: (310) 825-1144 (office) } } } } } } } } (310) 206-1029 (Lab) } } } } } } } } FAX (departmental): (310) 206-5272 } } } } } } } } mailto:sryazant-at-ucla.edu } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } _____________________________________ } } } } } } } } } } } } Sergey Ryazantsev Ph. D. } } } } } } Electron Microscopy } } } } } } UCLA School of Medicine } } } } } } Department of Biological Chemistry } } } } } } 10833 Le Conte Ave, Room 33-089 } } } } } } Los Angeles, CA 90095 } } } } } } } } } } } } Phone: (310) 825-1144 (office) } } } } } } (310) 206-1029 (Lab) } } } } } } FAX (departmental): (310) 206-5272 } } } } } } mailto:sryazant-at-ucla.edu } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } ------- End of forwarded message ------- } } } } } ========================================== } } } } } Dr. Chris Jeffree } } } } } University of Edinburgh } } } } } BIOSEM - Biological Sciences Electron Microscope Facility } } } } } Institute of Cell and Molecular Biology } } } } } Daniel Rutherford Building } } } } } King's Buildings, Mayfield Road } } } } } EDINBURGH, EH9 3JH, Scotland, UK } } } } } Tel. #44 (0) 131 650 5554 } } } } } FAX. #44 (0) 131 650 5392 } } } } } Mobile 07710 585 401 } } } } } email c.jeffree-at-ed.ac.uk } } } } } ========================================= } } } } } } } } _____________________________________ } } } } } } } } Sergey Ryazantsev Ph. D. } } } } Electron Microscopy } } } } UCLA School of Medicine } } } } Department of Biological Chemistry } } } } 10833 Le Conte Ave, Room 33-089 } } } } Los Angeles, CA 90095 } } } } } } } } Phone: (310) 825-1144 (office) } } } } (310) 206-1029 (Lab) } } } } FAX (departmental): (310) 206-5272 } } } } mailto:sryazant-at-ucla.edu } } } } } } } } } } } } } } } } } } } } } } } } } ========================================== } } } Dr. Chris Jeffree } } } University of Edinburgh } } } BIOSEM - Biological Sciences Electron Microscope Facility } } } Institute of Cell and Molecular Biology } } } Daniel Rutherford Building } } } King's Buildings, Mayfield Road } } } EDINBURGH, EH9 3JH, Scotland, UK } } } Tel. #44 (0) 131 650 5554 } } } FAX. #44 (0) 131 650 5392 } } } Mobile 07710 585 401 } } } email c.jeffree-at-ed.ac.uk } } } ========================================= } } } } _____________________________________ } } } } Sergey Ryazantsev Ph. D. } } Electron Microscopy } } UCLA School of Medicine } } Department of Biological Chemistry } } 10833 Le Conte Ave, Room 33-089 } } Los Angeles, CA 90095 } } } } Phone: (310) 825-1144 (office) } } (310) 206-1029 (Lab) } } FAX (departmental): (310) 206-5272 } } mailto:sryazant-at-ucla.edu } } } } } } } }
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
We are replacing our working Philips 410. It has been under continuous service contract since 1985. If interested in acquiring it, please contact me ASAP.
Thanks in advance
Steve ************************************************** * Join us for our annual * * * * INNERSPACE/OUTERSPACE OPEN HOUSE * * * * SATURDAY NOVEMBER 15, 2003 * * * * 4-8 PM , FREE ADMISSION * * * * www.sci.sdsu.edu/EM_Facility/ioflyer.html * * * ************************************************** Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619) 594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu http://www.sci.sdsu.edu/emfacility
Chairman, Educational Outreach subcommittee promoting microscopy instruction and increased access to microscopes Microscopy Society of America http://www.msa.microscopy.com/
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 16 01:18:11 2003
We also use Graphic converter as well as Corel Draw. Alby On Mercoledì, ott 15, 2003, at 17:55 Europe/Rome, Lesley Weston 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 } ----------------------------------------------------------------------- } -------- } } If you're using a Mac, there's an excellent shareware programme called } Graphic Converter, which can read anything and translate it into } anything } else. } } http://www.lemkesoft.com/ } } Hope this helps. } } Lesley Weston. } } } on 14/10/2003 3:44 PM, Ritchie Sims at r.sims-at-auckland.ac.nz 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 } } } ----------------------------------------------------------------------- } ----- } --} - } } } } } } Have you tried the good old IrfanView (free from www.irfanview.com)? } } } } cheers } } } } rtch } } } } } } Subject: [Microscopy] Image reader } } Date sent: Tue, 14 Oct 2003 16:09:02 -0500 } } } From: "Beavers, Roy" {rbeavers-at-mail.smu.edu} } } To: "Microscopy Listserver" } } {Microscopy-at-sparc5.microscopy.com} } } } } } } } } } } } --------------------------------------------------------------------- } } } - } } } -------- 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 } } } --------------------------------------------------------------------- } } } - } } } --------- } } } } } } Group, } } } } } } } } } I have recently collected some large (13.8M) 16 bit grayscale tiff } } } images from my SEM. From the SEM software they look very good and } } } print well to my inkjet printer. The problem I am having is reading } } } the files into any 3rd party imaging software (ie Photoshop, Photo } } } Impact) for resizing and saving in additional formats such as jpeg. } } } Is there software out there that can handle such files or am I } } } missing something about the tiff file format? } } } } } } Thanks } } } } } } Roy Beavers } } } Southern Methodist University } } } Department of Geological Sciences } } } P.O. Box 750395 } } } Dallas, Tx. 75275 } } } Voice: 214-768-2756 } } } Fax: 214-768-2701 } } } Email: rbeavers-at-mail.smu.edu } } } } } } } } } } } } } } } } } } } ------------------------------------------------------------------------ -------------------------- Alberto Diaspro Department of Physics, University of Genoa Via Dodecaneso 33, 16146 Genoa Italy voice +390103536426/480, facsimile +39010314218 visit http://www.lambs.it; http://www.wiley.com/WileyCDA/WileyTitle/productCd-0471409200.html ------------------------------------------------------------------------ -------------------------- RESISTER! RESISTERE! RESISTERE! ------------------------------------------------------------------------ --------------------------
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 16 03:12:31 2003
I was wondering if anybody of you know, where I could get a gold coated insect! It would be very nice to have such a toy when presenting our SEMs to freshers or pupils.
Regards Dirk Kirch
-- begin:vcard n:Kirch;Dirk tel;fax:+49(0)241-8022301 tel;work:+49(0)241-8026861 x-mozilla-html:FALSE url:www.imm.rwth-aachen.de org:Institute of Physical Metallurgy and Metal Physics;University of Aachen version:2.1 email;internet:kirch-at-imm.rwth-aachen.de title:Dipl.- Phys. Dirk Kirch adr;quoted-printable:;;Kopernikusstra=DFe 14=0D=0A;52056 Aachen;Germany;; end:vcard
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 16 03:40:28 2003
Sergey I was thinking about accuracy in densitometry applied to quantitative estimation of something, e.g. in cytochemistry, where density of the negative is calibrated to be equivalent to some amount of material. In quantitative autoradiography for example one would expect to have to control the exposure time and the development conditions - chemistry, concentration, temperature, time, agitation. But you should also standardise the conditions under which the negatives are fixed, and not leave them in the fix while you go off to the pub. Chris
ps round here most of the aggressive oxygen seems to be bound up in C2H5OH
} Hi Chris } I think, you guys have some special oxygen, more aggressive, I guess. } So, your butter should be softer and brain healthier. You right, it's } 7 levels of grey. I don't understand what you meant talking about } accuracy? Of coarse I did average for the area about 2cm^2 in both } control and treated with fixer. But still, I did not expect to have } even such effect. Thanks for the good point. Sergey. } } At 12:10 AM 10/15/2003, you wrote: } } Sergey } } I estimated the effect as a bit stronger than that, but maybe the } } oxygen concentration is greater in this part of the world. That could } } account for all sorts of stuff - the sheer quality of Scottish } } science, the frequency of Glasgow house fires... To be serious for a } } moment, 3-5% reduction may not always be important pictorially if it } } takes place uniformly, but that is unlikely to occur in a neglected } } fixer bath. Also converting the effect of 3-5% reduction into grey } } levels you might expect a density change of about -7 to -13 which } } won't contribute any accuracy to your densitometry. Chris } } } } Dr. Chris Jeffree } } Edinburgh University } } Biological Sciences EM Facility } } } } ----- Original Message ----- } } From: "Sergey Ryazantsev" {sryazant-at-ucla.edu} } } To: {Microscopy-at-sparc5.microscopy.com} } } Sent: Wednesday, October 15, 2003 3:29 AM } } Subject: [Microscopy] Re: Re: Fixer life SO163 Development } } } } } } } } } } } } } ------------------------------------------------------------------ } } } -- } } ---------- } } } 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 Chris } } } I think you are right. When I immersed half film in the fixer - } } after a } } } few hours I did see noticeable bleached line on the border between } } fixer } } } and air. This line is about 1 mm thick by the way. It clearly } } indicates } } } for me that air is involved (most noticeable on the water-air } } } interface). Except that interface line, the effect of bleaching } } } was } } about } } } 3% from untouched film for 1.5 h exposure to the fixer. My guess } } } is } } that } } } at fixer-air interface the metal Ag is more available for the } } oxidation } } } and then bleaching by the sodium thiosulphate as it correctly } } } cited } } in your } } } last posting. 3% bleaching effect inside the solution is not big } } deal: } } } variation in temperature/time perhaps will benefit more pronounced } } effect } } } on the film. But still, I am impressed: this is a pure example } } } how } } real } } } life interfere with our basic knowledge without any respect to } } } the "theory"... Anyway, thanks for pointing out this problem. } } } Sergey. } } } } } } At 02:16 AM 10/13/2003, 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 } } } } } } ------------------------------------------------------------------- } } } -- } } ---------- } } } } } } } } Sergey } } } } I don't pretend to be able to account for this effect, but it } } happens } } } } anyway. Try } } } } this simple test: stand an unwanted negative on edge with lower } } half immersed } } } } in a bath of your favourite acid or rapid fixer overnight. Wash } } } } and } } dry. } } } } Compare } } } } density of immersed and non-immersed areas. } } } } } } } } See quotes below from } } } } Grant Haist (1979) Modern Photographic Processing; John Wiley & } } Sons, } } } } Inc.; New York; Vol. 1; pps. 564-565 } } } } "Another danger of long fixing bath immersions is the direct } } } } attack } } on the } } } } silver image by the combination of oxygen and the acid fixing } } } } bath. } } It is } } } } believed that oxygen from the air dissolves in the fixing bath } } } } and } } attacks } } } } the } } } } very finely divided silver particles of the image. Oxygen } } } } converts } } these } } } } metallic silver particles to silver ions by removing electrons. } } } } The } } silver } } } } ions } } } } are then complexed by the thiosulfate and removed, resulting in } } } } the } } loss of } } } } image silver. Fine-grained films and paper images are especially } } susceptible } } } } to image reduction by acid thiosulfate solutions." } } } } "The extent of the reducing effect of fixing baths on the silver } } image during } } } } the process of fixation is greater than has been generally } } supposed. This } } } } attack does not occur in alkaline thiosulfate solutions" } } } } Best wishes } } } } Chris } } } } } } } } } } } } On 10 Oct 03, at 20:38, 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 } } } } } } } ------------------------------------------------------------------ } } } -- } } -- } } } } } --------- } } } } } } } } } } Chris } } } } } Thanks for your message. It forced me to refresh my memory in } } } } } chemistry. The process of dissolving of metal silver is } } reduction. } } } } } Metal Ag in the film is already reduced in compare with silver } } halide. } } } } } The conditions in the fixer are again reducing, so there is no } } } } } chemical direct way to reduce metal silver in the fixer. It } } meant that } } } } } directly sodium thiosulphate could not dissolve the metal } } } } } silver } } in } } } } } the film but silver halide. There are couple of things you } } } } } need } } to } } } } } keep in mind: if for some reason silver is oxidized, then you } } could } } } } } reduce it with sodium thiosulphate and therefore "dissolve } } } } } it". Because general reducing conditions in the fixer, oxygen } } } } } from } } air may } } } } } not be effective. You need to add something like bleach into } } the } } } } } fixer to oxidize silver and then you may reduce it with sodium } } } } } thiosulphate. Another thing comes to my mind is chlor gas. } } } } } So, } } you } } } } } may keep film in the fixer for quite while and silver will not } } } } } dissolved (if only you will not add the bleach). Professional } } } } } photographers usually do not recommend to keep film in fixer } } } } } for } } so } } } } } long for another reason: sodium thiosulphate is slowly } } decomposing } } } } } with creation of elementary sulphur, which colored film in } } yellowish } } } } } color. If you will keep your film in the fixer for couple of } } days, } } } } } elementary sulfur will be deposited in the gelatin layer, } } } } } which } } made } } } } } film yellowish irreversibly. Similar thing is happening when } } } } } you } } do } } } } } not wash film well after fixer - the residue of sodium } } thiosulphate } } } } } will decompose with sulphur creation. So, it's good practice } } } } } to } } ensure } } } } } your film is washed well after fixer. Have a great wekend, } } Sergey } } } } } } } } } } At 03:08 AM 10/10/2003, 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 } } } } } } } } ------------------------------------------------------------------- } } } -- } } } } } } ---------- } } } } } } } } } } } } Sergey } } } } } } Sodium thiosulphate has a mild solvent effect on silver, and } } this is } } } } } } accelerated at low pH in acid and hardening fixer } } } } } } formulations. } } The } } } } } } consequences are negligible during standard fixing times for } } film or } } } } } } paper, but prolonged over-fixing will produce noticeable } } bleaching, } } } } } } reducing overall negative or print density and bleaching out } } low } } } } } } density areas so that shadow detail in negatives and } } } } } } highlight } } detail } } } } } } in prints are lost. } } } } } } } } } } } } An old rule of thumb to determine the required fix time for } } } } } } EM } } film } } } } } } in old or partially-used fixer is twice the time taken to } } } } } } clear } } the } } } } } } film. Another rule of thumb is to discard the fixer when the } } clearing } } } } } } time is twice that for a fresh solution. Development is } } } } } } stopped } } very } } } } } } quickly in a fix bath unless it is completely exhausted, and } } } } } } it } } is } } } } } } perfectly safe to inspect the negatives as they clear in } } safelight or } } } } } } even in subdued white light. The most serious downside of } } stretching } } } } } } fixer life to the limit is that it increases the risk of } } } } } } silver staining of the negatives, and may reduce their } } } } } } archival } } permanence. } } } } } } } } } } } } I gave a formula for D19 on this list some months back. Here } } } } } } it } } is } } } } } } again, derived from British Journal of Photography almanac. } } } } } } } } } } } } Metol 2.2g, Hydroquinone 8.8g, Sodium sulphite 144g, Sodium } } carbonate } } } } } } 130g Potassium bromide 4g, H2O to 1litre } } } } } } } } } } } } However, other sources quote markedly different proportions } } } } } } of } } the } } } } } } ingredients, e.g. this one from Leica Users group } } } } } } http://mejac.palo-alto.ca.us/leica-users/v03/msg13172.html } } } } } } D19: } } water } } } } } } 500 cc Metol 2.0 grams sodium sulfite, desc. 90.0 grams } } Hydroquinone } } } } } } 8.0 grams sodium carbonate, monohyd. 52.5 grams potassium } } bromide 5 } } } } } } grams cold water to make 1.0 liter and this one from } } } } } } http://astro.umsystem.edu/apml/ARCHIVES/MAR01/msg01118.html } } } } } } You } } can } } } } } } make D19 yourself by using its formula: Metol 2 g } } Sodium } } Sulfite } } } } } } 90 g } } Hydroquinone 8 g } } Sodium Carbonate (anhydrous) 45 } } } } } } g } } } } } } } } Potassium Bromide 5 g } } Water 1 liter } } } } } } } } } } } } What is the official formula of Kodak D19 as currently } } marketed. } } } } } } Anyone have authoritative information? } } } } } } } } } } } } } } } } } } Dr. Chris Jeffree } } } } } } University of Edinburgh } } } } } } Biological Sciences EM Faciility } } } } } } } } } } } } ----- Original Message ----- } } } } } } } From: "Sergey Ryazantsev" {sryazant-at-ucla.edu} } } } } } } To: {Microscopy-at-sparc5.microscopy.com} } } } } } } Sent: Friday, October 10, 2003 4:49 AM } } } } } } Subject: [Microscopy] Re: SO163 Development } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } ------------------------------------------------------------------ } } } } } } } -- } } } } } } ---------- } } } } } } } 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 } } } } } } } } } ------------------------------------------------------------------ } } } } } } } -- } } } } } } ----------- } } } } } } } } } } } } } } Garry } } } } } } } You are right and aren't. Kodak spent a lot of time (and } } perhaps } } } } } } money) to } } } } } } } develop chemical formulations, which are stable over some } } period } } } } } } } of } } } } } } time } } } } } } } being in use. It, actually, made a revolution in the } } photographic } } } } } } process: } } } } } } } all C-41 process machines in our drug-stores used the } } solutions, } } } } } } which } } } } } } } stable for quite a while. It dramatically reduces the } } } } } } } cost } } of } } } } } } developing } } } } } } } and maintenance. Similarly they invented "long-play" } } solutions to } } } } } } develop } } } } } } } X-rays films in the hospitals (we do have such machine - } } } } } } } technician } } } } } } changed } } } } } } } solutions ones a month). Because in TEM it's just } } impossible to } } } } } } } use } } } } } } fresh } } } } } } } developer every time you need to develop a few films and } } Kodak } } } } } } suggest to } } } } } } } use "used" developer over some period of time, I suspect } } D-19 has } } } } } } "extended } } } } } } } life" formulation as well. Based on my personal } } } } } } } experience, } } it's } } } } } } clear to } } } } } } } me that diluted solution of D-19 (used or non-used) } } } } } } } deliver } } about } } } } } } the same } } } } } } } results for at least 3 weeks if properly stored without } } exposure } } } } } } } to air. Usually we develop about 100 films in 1.5 liter. } } } } } } } I think, the } } } } } } decent } } } } } } } quality of the TEM film developed in used developer is } } } } } } } also because } } } } } } of } } } } } } } special film formulation (bye-bye old 4489!). It's called } } } } } } } "electron microscopy film" and I assume it separates this } } film } } } } } } } from other type } } } } } } of the } } } } } } } films. As for price for chemicals - please count your } } technician } } } } } } time to } } } } } } } make fresh solutions 3 times a day and utilize used ones, } } it's not } } } } } } cheap as } } } } } } } you think. On the positive side, I do agree with you } } } } } } } that } } it's } } } } } } good idea } } } } } } } in general to use fresh components everywhere: in } } photography, } } } } } } } biochemistry, cooking etc. Sergey } } } } } } } } } } } } } } P.S. You don't need to change Stop-bath and fixer so } } } } } } } often. } } I do } } } } } } believe, } } } } } } } that Kodak suggests that stop-bath solution is good for } } 1000+ } } } } } } films... The } } } } } } } things about fixer - it could not spoil your film as long } } } } } } } as } } you } } } } } } careful do } } } } } } } not mix it with developer. You may left film in the fixer } } for } } } } } } } hours } } } } } } and it } } } } } } } does not harm film. You need to wash it after all by the } } way. } } } } } } } The } } } } } } bottom } } } } } } } line here is that you may extend fixation time if your } } fixer } } } } } } } start } } } } } } working } } } } } } } slowly saving money and doing good for environment (less } } chemical } } } } } } waste). } } } } } } } } } } } } } } } } } } } } } At 07:46 PM 10/9/2003, you wrote: } } } } } } } } While I don't use SO163 film or compatible developers, I } } } } } } } } disagree in general about the life of film developer mix. } } } } } } } } The film developer is the most important step in } } } } } } } } processing silver halide media. At least, this is how I } } } } } } } } see it. } } } } } } } } } } } } } } } } I use (or would have used) fresh developer for each } } } } } } } } batch of film that I developed. And Kodak explicitly } } } } } } } } details (as do others) how many rolls or sheets of film } } } } } } } } can be processed with a fresh batch before refreshing or } } } } } } } } dumping a pot of developer. I never refreshed developer. } } } } } } } } The results are just not the same as with a fresh batch. } } } } } } } } } } } } } } } } It may be that this TEM film is pathetically ignorant of } } } } } } } } basic photo chemistry. However, for such a small cost of } } } } } } } } chemicals, why take the chance? If the shots are worth } } } } } } } } shooting, they are worth proper processing. That means } } } } } } } } new chemicals. And, these chemicals must be at the } } } } } } } } correct temperature to be consistent and effective. I've } } } } } } } } seen too many instances of no temperature control. } } } } } } } } Consequently, the results vary wildly. Duh. } } } } } } } } } } } } } } } } Furthermore, I dumped stop bath and fixer routinely. Why } } } } } } } } cut corners after so much work on the instrument? } } } } } } } } } } } } } } } } gary g. } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } At 05:12 PM 10/9/2003, 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 } } } } } } } } } } } } } } } } } ------------------------------------------------------------------ } } } } } } } } -- } } } } } } ----------- } } } } } } } } } } } } } } } } } } Basically, the life of unused (even diluted) developer } } } } } } } } } is } } quite } } } } } } long } } } } } } } } } (without air). The clock starts ticking as soon as you } } develop } } } } } } even one } } } } } } } } } piece of the film (similar but less dramatic for paper } } } } } } developers). The } } } } } } } } } amount of film company recommends to develop usually } } } } } } } } } mean } } that } } } } } } } } } you supposed to develop all those films in relatively } } short } } } } } } } } } period of time. Classical example is quite popular in } } amateur } } } } } } } } } photography } } } } } } D-76 } } } } } } } } } developer. In 1 liter of the fresh developer you could } } develop } } } } } } about 5 } } } } } } } } } rolls of 35 mm film, but developer will die in about 4-5 } } hours } } } } } } does not } } } } } } } } } matter how many films you manage to develop, so hurry } } } } } } } } } up! } } } } } } Similarly, } } } } } } } } } it's true for D-19 especially non-diluted. It will not } } die in } } } } } } } } } few } } } } } } hours, } } } } } } } } } but perhaps two weeks. The exposure to air is very } } critical } } } } } } } } } here also. So, the bottom line here is: if you need to } } develop } } } } } } } } } a lot } } } } } } of films } } } } } } } } } in quite short period of time - you may do it to extend } } } } } } } } } company's recommendation. As soon as you start use } } developer, it } } } } } } } } } will } } } } } } gradually } } } } } } } } } lost capacity in the matter of few weeks and will die } } } } } } } } } even } } if } } } } } } } } } you } } } } } } just } } } } } } } } } developed 10 films. Note: this does not applied to the } } fixer - } } } } } } } } } as } } } } } } long } } } } } } } } } as you do not contaminate it with developer, it will } } } } } } } } } works } } to } } } } } } } } } the } } } } } } end of } } } } } } } } } capacity, does not matter how long it will take (6 mo } } } } } } } } } for } } sure } } } } } } } } } and } } } } } } even } } } } } } } } } longer). Fixer will die soon if you contaminate it with } } } } } } developer. By } } } } } } } } } the way - if you are using those test drops to check } } fixer - } } } } } } } } } they } } } } } } are too } } } } } } } } } sensitive, you may use fixer much longer and for more } } films } } } } } } without any } } } } } } } } } harm to the film. Experimentally I determined that when } } those } } } } } } "drops" } } } } } } } } } indicate, the fixer is bad, it actually used about 50% } } } } } } } } } of } } } } } } capacity. I } } } } } } } } } hope it helps. Sergey } } } } } } } } } } } } } } } } } } } } } } } } } } } At 06:11 AM 10/9/2003, 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 Leslie: } } } } } } } } } } } } } } } } } } } } 12 minutes in straight D-19 sounds like a huge push } } to me! } } } } } } } } } } On } } } } } } the } } } } } } } } } } other hand, I have no experience with cryo EM so ... } } Have you } } } } } } talked to } } } } } } } } } } Kodak about this? Long times in developer can lead to } } fogging. } } } } } } } } } } As to life of the chemistry in a tank. If you have } } } } } } } } } } a floating } } } } } } lid } } } } } } } } } } (that excludes most air) the chemistry will keep } } } } } } } } } } better } } than } } } } } } } } } } if } } } } } } the } } } } } } } } } } lid allows free access to air. Again, a call to Kodak } } is in } } } } } } order. As } } } } } } } } } } for the number of films I always stay on the } } conservative } } } } } } } } } } side, } } } } } } maybe } } } } } } } } } } 2/3 of the recommendation. } } } } } } } } } } D-19 is the cheapest part of the equation (animals, } } prep } } } } } } time, etc) } } } } } } } } } } so trying to squeez a few extra films out of a pot of } } } } } } } } } } developer } } } } } } is not } } } } } } } } } } the way to go. Also, D-19 does not last more than 2 } } years in } } } } } } } } } } the package on the shelf. I don't do much TEM anymore } } } } } } } } } } so } } I mix } } } } } } } } } } my } } } } } } D-19 from } } } } } } } } } } scratch using the individual chemicals, not the } } prepackaged } } } } } } Kodak mix. } } } } } } } } } } The recipe is widely published, Kodak will give it to } } you if } } } } } } } } } } you } } } } } } ask. } } } } } } } } } } } } } } } } } } } } Geoff } } } } } } } } } } } } } } } } } } } } Leslie Cummins wrote: } } } } } } } } } } } } } } } } } } } } } Hello Listers } } } } } } } } } } } } } } } } } } } } } } We are setting up to develop Kodak SO163 film for low } } dose } } } } } } cryoEM. We } } } } } } } } } } } are push processing it, so we are developing it for 12 } } minutes } } } } } } in } } } } } } } } } } } straight D-19, we're using a nitrogen gas burst, for 1 } } second } } } } } } every 8 seconds. } } } } } } } } } } } } } } } } } } } } } } I was wondering what the life of the chemistry might } } } } } } } } } } } be. Kodak } } } } } } said } } } } } } } } } } } that we could develop 60 8x10 negatives per gallon. } } This } } } } } } } } } } } works } } } } } } out to } } } } } } } } } } } about 360 3 1/4 x 4 negatives. This seems like a lot } } } } } } } } } } } of } } } } } } negatives to } } } } } } } } } } } us, so we are wondering what other people are using } } } } } } } } } } } for } } an } } } } } } exhaustion } } } } } } } } } } } time, both in negative numbers and days in the tank. } } } } } } } } } } } } } } } } } } } } } } Thanks for the help. } } } } } } } } } } } Leslie Cummins } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } Leslie Gunther Cummins } } } } } } } } } } } Analytical Imaging Facility } } } } } } } } } } } Albert Einstein College of Medicine } } } } } } } } } } } 1300 Morris Park Ave. } } } } } } } } } } } Bronx, NY 10461 } } } } } } } } } } } 718-430-3547 } } } } } } } } } } } } } } } } } } } } } } http://www.aecom.yu.edu/aif/ } } } } } } } } } } } } } } } } } } } } -- } } } } } } } } } } -- } } } } } } } } } } ********************************************** } } } } } } } } } } 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 } } } } } } } } } } ********************************************** } } } } } } } } } } } } } } } } } } } } } } } } } } } } _____________________________________ } } } } } } } } } } } } } } } } } } Sergey Ryazantsev Ph. D. } } } } } } } } } Electron Microscopy } } } } } } } } } UCLA School of Medicine } } } } } } } } } Department of Biological Chemistry } } } } } } } } } 10833 Le Conte Ave, Room 33-089 } } } } } } } } } Los Angeles, CA 90095 } } } } } } } } } } } } } } } } } } Phone: (310) 825-1144 (office) } } } } } } } } } (310) 206-1029 (Lab) } } } } } } } } } FAX (departmental): (310) 206-5272 } } } } } } } } } mailto:sryazant-at-ucla.edu } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } _____________________________________ } } } } } } } } } } } } } } Sergey Ryazantsev Ph. D. } } } } } } } Electron Microscopy } } } } } } } UCLA School of Medicine } } } } } } } Department of Biological Chemistry } } } } } } } 10833 Le Conte Ave, Room 33-089 } } } } } } } Los Angeles, CA 90095 } } } } } } } } } } } } } } Phone: (310) 825-1144 (office) } } } } } } } (310) 206-1029 (Lab) } } } } } } } FAX (departmental): (310) 206-5272 } } } } } } } mailto:sryazant-at-ucla.edu } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } ------- End of forwarded message ------- } } } } } } ========================================== } } } } } } Dr. Chris Jeffree } } } } } } University of Edinburgh } } } } } } BIOSEM - Biological Sciences Electron Microscope Facility } } } } } } Institute of Cell and Molecular Biology Daniel Rutherford } } } } } } Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, } } } } } } Scotland, UK Tel. #44 (0) 131 650 5554 FAX. #44 (0) 131 650 } } } } } } 5392 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk } } } } } } ========================================= } } } } } } } } } } _____________________________________ } } } } } } } } } } Sergey Ryazantsev Ph. D. } } } } } Electron Microscopy } } } } } UCLA School of Medicine } } } } } Department of Biological Chemistry } } } } } 10833 Le Conte Ave, Room 33-089 } } } } } Los Angeles, CA 90095 } } } } } } } } } } Phone: (310) 825-1144 (office) } } } } } (310) 206-1029 (Lab) } } } } } FAX (departmental): (310) 206-5272 } } } } } mailto:sryazant-at-ucla.edu } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } ========================================== } } } } Dr. Chris Jeffree } } } } University of Edinburgh } } } } BIOSEM - Biological Sciences Electron Microscope Facility } } } } Institute of Cell and Molecular Biology } } } } Daniel Rutherford Building } } } } King's Buildings, Mayfield Road } } } } EDINBURGH, EH9 3JH, Scotland, UK } } } } Tel. #44 (0) 131 650 5554 } } } } FAX. #44 (0) 131 650 5392 } } } } Mobile 07710 585 401 } } } } email c.jeffree-at-ed.ac.uk } } } } ========================================= } } } } } } _____________________________________ } } } } } } Sergey Ryazantsev Ph. D. } } } Electron Microscopy } } } UCLA School of Medicine } } } Department of Biological Chemistry } } } 10833 Le Conte Ave, Room 33-089 } } } Los Angeles, CA 90095 } } } } } } Phone: (310) 825-1144 (office) } } } (310) 206-1029 (Lab) } } } FAX (departmental): (310) 206-5272 } } } mailto:sryazant-at-ucla.edu } } } } } } } } } } } } } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } 10833 Le Conte Ave, Room 33-089 } Los Angeles, CA 90095 } } Phone: (310) 825-1144 (office) } (310) 206-1029 (Lab) } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu } } } }
========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 (0) 131 650 5554 FAX. #44 (0) 131 650 5392 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 16 04:45:03 2003
Thank you for your answer. I try to explain what I want. Firstly, I have not hematite in my systems. I intend to prepare ferrihydrite in the presence of smectite. Some researches showed the Fe (and Al also) may be enter in different forms in the smectite interlayer, leading to as called hydroxy-Fe interlayered smectites. Thus, the reactions between disordered Fe oxides and smectites are complex, because these latter may be adsorbed also on the external surface of smectite. The XRD measurements may indicate the d spacing variation, but it would be possible that these Fe complexes enter into the smectite interlayer as islands, not as continuous layer. XRD can indicate in this case some peak modifications, but not details. I need TEM to see how it is happening. I would need to have an image of the interaction extension. I also try to have a chemical information from these smectite crystals. I intend to use L.R.White resin to maintain the original interlayer spacing to compare the crystals between them. I do not know if now I was clear enough. Maybe I expect too much from TEM; but I would wish to try. I already know that the centrifugation gives a differential settling of the phases. For this reason I wanted to try another method, but for moment I have not choice. The filtration would be the best, but it was impossible to obtain something. Maybe the pomp is not strong enough or the clay expansion and impermeability stops the process. Now I am trying to use the centrifugation, but I put each rotation time a small quantity to obtain 2-3 solid beds. Thus I am trying to keep in the sample all the phases. I know that this process will disturb also the relations between the mineral phases. For this reason I asked help. If I will discover at the end that my intention is utopical, I will have to change my idea. Now I am still waiting, hopping to find something.
Thank you again
----------------------------------------------------------------------------------- Prof. Elena BELLUSO - Ph.D. Mineralogy and Crystallography Dipartimento di Scienze Mineralogiche e Petrologiche Università degli Studi di Torino Via Valperga Caluso, 35 I-10125 TORINO - ITALIA tel: (39) 011 670 71 35 - fax: (39) 011 670 71 28 e-mail: elena.belluso-at-unito.it {http://www.dsmp.unito.it/} http://www.dsmp.unito.it
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From MicroscopyL-request-at-ns.microscopy.com Thu Oct 16 05:33:34 2003
} Most imaging applications do not support 16-bit TIFF files, including } Adobe Photoshop ...
Not true ... PS has been supporting 16bit (and 48bit) TIFFs since version 4. Granted, not every tool is available, but all tonal adjustments and noise removal is supported.
} You could use our software,
Of course! ... {TIC} ...
cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 16 06:46:04 2003
My mistake. The current version of Photoshop supports reading 16-bit and 48-bit TIFF files. Maybe the application you have uses some special tags in the header that Photoshop doesn't recognize. Often times special lookup tables are used and embedded. Great liberties are taken with TIFF (the TIFF specification allows for great flexibility) and there is no organization that certifies that you write a genuine TIFF file.
Dean Sequera
-----Original Message----- } From: Dean Sequera [mailto:DSequera-at-mediacy.com] Sent: Wednesday, October 15, 2003 2:37 PM To: 'Microscopy Listserver'
I agree with Michael, Photoshop does support 16-bit TIF images. However, we ran into this problem a while ago: A user of our software could open a TIF image acquired with analySIS in PS, but it was all black. I don't know if this is the same problem, but the solution to the problem was fairly easy. A bit of background:
There are very few b/w cameras that actually provide 16 bit of data. Most cameras are 12 bit or 14 bit. As there is no 12 or 14 bit data format for storing (they come in multiples of 1 byte=8bit), a 12 bit image has to be stored as a 16 bit file. 16 bit span the range of 0 to 65,536, but the data only span 0 to 4,096. If PS does not do an automatic adjustment, the 12-bit data is displayed as very low intensity and the image appears black.
The solution to the problem was to simply have PS do a "histogram stretch" (I think that was the term PS uses, but I am not sure. I don't really use PS) and the image appeared in all its glory.
There is of course also a possibility that there is something wrong with the TIF file. If the original poster is still listening: If you want to send me the file I could try to open it in our software and convert it for you. Please contact me offline for instructions (mb-at-soft-imaging.com).
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: michael shaffer [mailto:michael-at-shaffer.net] Sent: Thursday, October 16, 2003 4:33 AM To: Dean Sequera Cc: Microscopy Listserver
Dean Sequera (of MediaCybernetics) writes ...
} Most imaging applications do not support 16-bit TIFF files, including } Adobe Photoshop ...
Not true ... PS has been supporting 16bit (and 48bit) TIFFs since version 4. Granted, not every tool is available, but all tonal adjustments and noise removal is supported.
} You could use our software,
Of course! ... {TIC} ...
cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 16 12:02:35 2003
Hi, 1) Does anyone know the name of the company that makes the OV60 crystal and might you have a phone number, email address, or url for them? 2) What is the typical delivery pressure for P-10 gas to WDS detectors?
TIA Mike -- ******************************************************************** Michael M. Cheatham 312 Heroy Geology Laboratory Phone (315)-443-1261 Syracuse University Fax (315)-443-3363 Syracuse, NY 13244-1070
owner of PLASMACHEM-L: http://listserv.syr.edu/archives/plasmachem-l.html owner of XRF-L: http://listserv.syr.edu/archives/xrf-l.html owner of TIMS-L: http://listserv.syr.edu/archives/tims-l.html owner of SIRIS-L: http://listserv.syr.edu/archives/siris-l.html ********************************************************************
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 16 12:17:38 2003
Dear Dirk, Most people who run SEM labs have a gold sputtering machine to coat non-conductive samples with a thin layer of gold or gold alloy. Samples that go into a conventional SEM must be electrically conductive. If I want to view an insect, I first dry it for two or three months, so that all the moisture inside is gone (otherwise the insect may explode when you try to pump it in a vacuum), then I cut it and mount it on an SEM stub so the eyes are up, then gold coat it several times. I have several that I use for school tours and have had them for years. I like to show the students the difference between insects and spiders. Regards,
----- Original Message ----- } From: "Dirk Kirch" {kirch-at-imm.rwth-aachen.de} To: "Microscopy" {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, October 16, 2003 1:14 AM
Yes, Photoshop can open 16-bit images, but they will appear black. To get initial control of them, go to Image - Adjust - Auto Levels, then you can work with the histogram from there. You may have to change them to 8-bit for many Photoshop functions. We have had no problems with Photoshop handling the 16-bit (14-bit) images generated with our analySIS software on our EFTEM.
However, the images generated from our analySIS ADDA II digital image acquisition system on our FESEM (changed to 8-bit) do not not open with some other programs, such as LViewPro, because of header issues. It was easy to have our user who used LViewPro switch to another progam (Irfanview) that could handle the header. The advantage of these programs is that they open the image much faster than Photoshop if you just want to take a look.
The header issue and the 16-bit issue are two different things. I think.
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 Thu Oct 16 14:50:11 2003
You might try SemTech Solutions of Billerca, MA. TN is (978) 663-9822 and on the web at www.semtechsolutions.com. They are an SEM service company that specializes in Amray scopes. If they cannot help you themselves they will probably know what direction to point you in.
Good luck,
Michael R. Nesta General Manager Energy Beam Sciences, Inc. Tel: 413 786-9322 Fax: 413 789-2786 mnesta-at-ebsciences.com www.ebsciences.com "Adding Brilliance to Your Vision"
-----Original Message----- } From: by way of Ask-A-Microscopist [mailto:scott.watson-at-granite.k12.ut.us] Sent: Monday, October 13, 2003 6:20 PM To: microscopy-at-ns.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (scott.watson-at-granite.k12.ut.us) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Monday, October 13, 2003 at 17:11:52 ---------------------------------------------------------------------------
Email: scott.watson-at-granite.k12.ut.us Name: Scott Watson
Organization: Hunter High School
Education: 9-12th Grade High School
Location: West Valley City, Utah
Question: Hello. I am the electronics, engineering, and scanning electron microscopy instructor at Hunter High School in West Valley City, Utah. Here at Hunter we have an old Amray 1200C2 SEM which has been extensively modified since Amray went out of business. The microscope has had 99.9 percent of electronic circuitry replaced with new/redesigned circuitry which transformed the SEM into an ESEM. Recently several of the universal swivel joints used for adjusting the X,Y,Z, and T axises of the specimen chamber have broken. I was wondering if you knew of a source for these joints? I only need to find a source for joints - they do not have to be original manufacturer? Do you know if another SEM manufacturers specimen chambers were similar in design to the Amray 1200 and could have their specimen axis adjustment joints placed into the Amray?
I have not had problems with Adobe Photoshop (R) handling 16 bit images in tif format - it works quite well even with images over 100MB. The problems I've seen arise when people attempt to save 16 bit images in jpg format.
When you first open your image in Photoshop (R), go into "Image" "Mode" and change your image from 16 bit to 8 bit. You should now be able to save your grayscale image in jpg format with no problem. It is also worth checking to be sure you really have a grayscale image - my SEM digitizing software defaults to an indexed color image which must be converted to grayscale, RGB, or CMYK before it can be saved as a jpg.
If Photoshop (R) complains about space on your computer, one of the first things to try is defragmenting the hard drive. Also check how you allocated space for Photoshop (R) when you set up the program.
You may also want to check that your files really are in tif format. I came across one program that "converted" tif to jpg by changing the extension to ".jpg" but did nothing else - needless to say the file size didn't change and other programs would not open the renamed file!
Good luck!
- Louise
(no ties to Adobe - just a happy user)
Louise Harner Research Microscopist Albany International Research Co. 777 West Street, P.O. Box 9114 Mansfield, MA 02048 508-337-9529 phone 508-339-4996 fax Louise_Harner-at-albint.com
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 16 16:25:31 2003
All, I am interested in your suggestions for vendors of phase separators for application in LN2 manual fill stations.
The last two products we used failed prematurely. In case of Cu sponge pressed on a SS fitting, use for about a month lead to Cu sponge increasing in diameter to fall of the fitting. In case of SS sponge welded to SS fitting, the weld heat affected zone in the sponge fails due to brittleness of the material during the first few fills (we might be contributing to it by dropping the hose from time to time). I am at the end of my rope. EHS requires us to have such devices installed.
Warm regards from Texas.
Jerzy
****************************************************** Jerzy Gazda, Ph.D. Advanced Micro Devices Supervising Engineer 5204 E. Ben White Blvd. - MS 512 PCAL - AIM Section Austin, TX 78741 TEL: 1-800-538-8450, Ext. 51453 jerzy.gazda-at-amd.com ******************************************************
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 16 18:26:20 2003
Does anyone know where I can lay my hands on an anode assembly for an S360? I have a friends who needs one. It must be cheap.
-- John Mansfield PhD MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 USA Phone: (734) 936-3352 FAX (734) 763-2282 Cell. Phone: (734) 834-3913 (Leaving a phone message at 936-3352 is preferable to 834-3913) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42! 16' 48" Long. 83! 43' 48"
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 16 18:48:41 2003
We've done this routinely for our visiting students and when we do the school tours. The drying time can actually be much quicker by placing the insect, etc. into a vial (although often we can find them already deceased)..
I've observed many of these creatures without the coating as well..simply using a low voltage can often work very well. For the higher magnifications, however, coating does result in better resolution.
Dirk, if you'd like some examples of the images, I can forward them over to you (let me know your preferred format).
Carol Jean Hirt Vice President Materials Research Laboratories, Inc. mrllab.com
} From: "Mary Mager" {mager-at-interchange.ubc.ca} } Date: Thu, 16 Oct 2003 10:15:59 -0700 } To: "Dirk Kirch" {kirch-at-imm.rwth-aachen.de} } Cc: "Microscopy" {microscopy-at-MSA.microscopy.com} } Subject: [Microscopy] Re: coated insects } } } } ------------------------------------------------------------------------------ } 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 Dirk, } Most people who run SEM labs have a gold sputtering machine to coat } non-conductive samples with a thin layer of gold or gold alloy. Samples that } go } into a conventional SEM must be electrically conductive. If I want to view an } insect, I first dry it for two or three months, so that all the moisture } inside } is gone (otherwise the insect may explode when you try to pump it in a } vacuum), } then I cut it and mount it on an SEM stub so the eyes are up, then gold coat } it } several times. I have several that I use for school tours and have had them } for } years. I like to show the students the difference between insects and spiders. } Regards, } } ----- Original Message ----- } } From: "Dirk Kirch" {kirch-at-imm.rwth-aachen.de} } To: "Microscopy" {Microscopy-at-sparc5.microscopy.com} } Sent: Thursday, October 16, 2003 1:14 AM } Subject: [Microscopy] coated insects } } } } } } } } ---------------------------------------------------------------------------- -} } - } } 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 Everybody! } } } } I was wondering if anybody of you know, where I could get } } a gold coated insect! It would be very nice to have such } } a toy when presenting our SEMs to freshers or pupils. } } } } Regards Dirk Kirch } } } } -- } } begin:vcard } } n:Kirch;Dirk } } tel;fax:+49(0)241-8022301 } } tel;work:+49(0)241-8026861 } } x-mozilla-html:FALSE } } url:www.imm.rwth-aachen.de } } org:Institute of Physical Metallurgy and Metal Physics;University of Aachen } } version:2.1 } } email;internet:kirch-at-imm.rwth-aachen.de } } title:Dipl.- Phys. Dirk Kirch } } adr;quoted-printable:;;Kopernikusstra=DFe 14=0D=0A;52056 Aachen;Germany;; } } end:vcard } } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 16 19:11:16 2003
I have an Amray 1700 Turbo that probably has the same stage mechanism. I have replaced several of the universal joints with ones purchased from Small Parts, Inc. www.smallparts.com. They offer delrin u-joints that fit the splines in my Amray at about $18. ea. They also sell the telescoping joints that appear identical to those on my stage.
Disclaimer: Edison Labs is a private analytical laboratory and has no interest in Small Parts other than as a satisfied customer.
James Carnahan
Edison Analytical Laboratories, Inc. 301 Nott Street Schenectady, NY 12305 (518) 393-2112
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 16 20:54:21 2003
I'm just getting into using my WDS system on my JXA-840A, setting up to look for low-level Zr in garnets, and I'm a bit disappointed with the apparent detection limits.
I'm using the Zr La, PET, 15kv, 15 nA beam current, 30 seconds on background, 100 seconds on peak, and 10 replicate analyses of a reputedly zero-Zr almandine garnet gave me mean 0.10% ZrO2, Excel STDEV 0.07.
If we take detection limit (whatever that means) as 3SDs, this is 0.21% ZrO2. I had hoped for better.
Other zero-concentartion elements gave (30 sec on peak, 10 sec on backgrounds) STDEVs of
These values are not a heck of an improvement on what I can get with EDS for 100- second counts at 1.5nA.
Is this about par for the WDS course?
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 16 21:58:32 2003
Can anyone supply me with useful information regarding the processing and sectioning of aortic stents embedded in plastic. I have tried several different resins (LX112 and LR White Grade Hard) as well as glass and diamond knifes with no success.
Thank you for your assistance. Sincerely,
Ken _______________________________________ Kenneth L. Tiekotter, Adjunct Professor The University of Portland Department of Biology 5000 N Willamette Blvd. Portland, OR 97203 USA
Director, MicroImaging Dx Center Legacy Portland Hospitals Legacy Holladay Park Medical Center 1225 NE 2nd Avenue Portland, OR 97232 USA
Tel.: 503.413.5391
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 17 06:39:43 2003
} ... A user of our software could open a TIF } image acquired with analySIS in PS, but it was all black. } I don't know if this is the same problem, but the solution } to the problem was fairly easy. ...
Yes ... maybe its because because I work with x-ray counts, but this should be expected. I'll always acquire x-ray counts with, and write to, 16bits, but many times I'll get less than 500 counts max. This will be a very dark 16bit TIFF, and could be 'equalized' for presentation, but it would no longer be 'real' data.
In defense of Cybernetics & Dean Sequera, Image Pro Plus definitely provides some advantages. It can equalize for presentation without changing the data, and as Dean made me aware (personal conversation), Cybernetics has investigated many of the irregular TIFF tags and headers, and can open them properly. Clearly, some TIFFs cannot be opened properly with Photoshop at all ... you may get to see the data, but you may not be able to interpret it.
SEM images are normal grayscale images however, and irregular TIFF tags and headers are a real problem. Whenever we detect a problem we should insist the manufacturer fix it. An example would be to include the definition of magnification without also including the print size. Manufacturers vary, but I have worked with their post-acquisition software and found it totally inadequate for ending up with presentation quality prints ... they really should acknowledge many of us would prefer to use something different ... and write TIFF files we can use.
I would like to prepare some Giemsa stained blood smears and preserve them by mounting with a coverslip. I am told that when using mounting media (such as Canada balsam, per mount, poly-mount, poly-mount-Aqua etc.) there is a chance that the stain may leach into the medium, or the medium bleach out the stain. Does anyone have a suggestion or experience with this stain? Is there a way other than empirically testing, to determine the best mounting medium to use? Thank you
W. Roy Prescott HydasR Inc. PO Box 420 Hershey, PA 17033 FAX - +1-717-533-5548 Voice - +1-717-533-5583 Mobile - +1-717-554-2572 email(s) Roy-at-Hydas.com Info-at-Hydas.com
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 17 09:59:07 2003
I found this out and fixed it easily by opening the TIFF file in Photoshop and then performing an Image/Auto Levels or Image/Auto Contrast. Auto Levels fixes the dynamic range of the smaller number of bits to match full 16-bits. Image then looks just like the original captured image.
gary g.
At 10:00 AM 10/16/2003, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 17 10:12:56 2003
Scott, There are small u-joints available through Berg www.wmberg.com and Reid Tool http://reidtool.com
You can also make your own with brass stock found in hobby shops. Square tubing stock is easier, although I believe AMRAY may have used round. The end can be ground to leave two ears sticking up about 3/16” on opposite sides. Drill a hole in each one. Cut a short (1/16” to 3/32”) piece of the next smaller size solid brass stock. Drill 2 holes all the way through, large enough to either accept small brass brads or tapped for 0-80 screws. Drill a hole (up to 1/8”) through the center where the previous holes intersect. If you use brass brads, put them each through one of the ears and into one of the holes in the “spider”. Trim the length of each so that they all meet in the center, where you drilled the larger hole, and solder them together. If you opted for screws, then trim four 0-80 brass or stainless screws so they will meet in the center and jam each other.
One of the chief advantages of this is that they can be repaired in the future, particularly easily if you opted for the screws.
Ken Converse owner QUALITY IMAGES 16 CREEK RD DELTA PA 17314 717-456-5491 FAX 717-456-7996 qualityimages-at-netrax.net
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 17 10:47:48 2003
you are now getting to the heart of the matter: TIF, as flexible as it may be, was not developed with microscopists in mind. Rather, the standard tags refer to parameters that a printer requires for a good rendering of the image. One tag you mention in your response:
"An example would be to include the definition of magnification without also including the print size."
First of all, magnification is not a very useful number. Let's say you print something at 100x and write the mag on the image. Next you take it to a copier and reduce it's size to 50%. The copy will still say 100x, but the image is only half the size as before, so it can't be 100x. Calibration is a much better way to do this. If you have a calibration bar, it scales with the image. So, magnification without specifying the image size is dangerous. Also, as far as I know, there is no public tag in the TIF format for magnification. You would have to convince whoever is responsible for the TIF format to put that in. That takes us to Calibration. There is actually a tag for that in TIF, but again, it is used differently than you'd expect. We had used that tag in the beginning to store image calibration (for example 1nm/pixel). That worked fine, until we tried to import the images into something like Word or other software. There the calibration value was interpreted as print size, so a 1kx1k image with a calibration value of 1nm/pixel was then interpreted to be printed with a size of 1 micron x 1 micron! Not very easy to see!!
What I want to say is, that even with the TIF format, as flexible as it is, we have to use non-standard headers in order to be able to use it.
Having said that, the TIF format does anticipate this and says, that "unknown" tags will be ignored by the standard. So, everybody can modify the header without interfering with the ability of other software to read the file, if done correctly. But it depends on both the writing software AND the reading software to adhere to the TIF formats. If they don't, all bets are off.
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 ===================================
An example would be to include the definition of magnification without also including the print size.
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 17 10:50:28 2003
On Thursday, October 16, 2003, at 02:25 PM, jerzy.gazda-at-amd.com wrote:
} I am interested in your suggestions for vendors of phase separators } for application in LN2 manual fill stations. } Dear Jerzy, I am not a vendor, but perhaps a simple solution would be to suspend a funnel from the fill line so the LN2 passes through the funnel while the gas does not. This worked well in many instances. Be sure to use a material that does not get brittle at 77 K, such as teflon. Good luck satisfying EHS. Yours, Bill Tivol 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 17 14:46:39 2003
I have used the same brass phase separator for over 14 years. I bought it at Liquid Carbonics, 1600 Perry Dr. SW, Canton, Ohio. The phone number then was 766-6900. The area code is probably 330 now. A lady named Glenda gave me part number LX0369; phase separator brass; 1-1/4" OD; 3/8" female NPT Nut. It looks like a pressed and sintered brass frit device. At the bottom of the order it says, "453-9904 Placed w/ plant but Canton will invoice".
The cost on April 3, 1989 was $38.50 but I am sure they cost more now. I measured the total length and it is 3.25 inches including the welded nut. The frit is a right circular cylinder 3 inches long.
This thing was real hard to get after someone broke my first one in 1989. For some reason Carbonics didn't like to bother with ordering them. I am sure Liquid Carbonics is another company by now. That's your problem.
A previous separator only failed when people dropped the thing onto concrete and cracked the bottom of the frit. I now take it with me--every time.
As for a funnel, I am using the same two funnels I bought from Fisher Scientific. They are polypropylene, Fisher number 10-371-F. None has ever exploded as was stated on the list awhile back. PE may be a problem but PP has worked for me for over 15 years. No warranty given.
Hope this helps you out and you find a source.
Sincerely,
Paul Beauregard Senior Research Associate PPG Industries Monroeville Technical Center 440 College Park Drive Monroeville, PA 15146
-----Original Message----- } From: jerzy.gazda-at-amd.com [mailto:jerzy.gazda-at-amd.com] Sent: Thursday, October 16, 2003 5:26 PM To: Microscopy-at-MSA.Microscopy.Com
All, I am interested in your suggestions for vendors of phase separators for application in LN2 manual fill stations.
The last two products we used failed prematurely. In case of Cu sponge pressed on a SS fitting, use for about a month lead to Cu sponge increasing in diameter to fall of the fitting. In case of SS sponge welded to SS fitting, the weld heat affected zone in the sponge fails due to brittleness of the material during the first few fills (we might be contributing to it by dropping the hose from time to time). I am at the end of my rope. EHS requires us to have such devices installed.
Warm regards from Texas.
Jerzy
****************************************************** Jerzy Gazda, Ph.D. Advanced Micro Devices Supervising Engineer 5204 E. Ben White Blvd. - MS 512 PCAL - AIM Section Austin, TX 78741 TEL: 1-800-538-8450, Ext. 51453 jerzy.gazda-at-amd.com ******************************************************
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 17 15:57:17 2003
Dear Ritchie, The formula I use for detection limit on my WDX system is: C1 = Cst x(3x(square root B)/P-B) where C1 is the detection limit in mass % Cst is the mass concentration of the analysed element in the standard (100% in Zr metal standard = 1, less if you use a compound standard) B is the background counts P is the peak counts. Make sure you normalize the peak and background counts to the same number of seconds or convert them to counts per second. As soon as you take your standard, you can calculate this. I don't know if this formula will give you a better or worse detection limit, but the WDX detection limits vary a lot more than EDX, because of the position of the different elements around the Roland Circle and how close to the sample the detector is for that particular element. You can then experiment with kV and nA settings to try to optimize the detection limit. Also, the other elements you tested were K lines and the Zr is an L line; this will also reduce the height of the peak and therefore the detection limit. Good luck, Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} To: {Microscopy-at-MSA.Microscopy.Com} Sent: Thursday, October 16, 2003 6:56 PM
Jerry, I can't tell you where to get a good phase separator, but there are also good economic reasons for having one: you will get far more of the liquid into your container with one than without. It pretty much eliminates the atomization that occurs without one.
In my experience, a sintered bronze separator should be available and work well for years.
Ken Converse owner Quality Images third party SEM service Delta, PA
Bill Tivol 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 } ------------------------------------------------------------------------------- } } } } On Thursday, October 16, 2003, at 02:25 PM, jerzy.gazda-at-amd.com wrote: } } } I am interested in your suggestions for vendors of phase separators } } for application in LN2 manual fill stations. } } } Dear Jerzy, } I am not a vendor, but perhaps a simple solution would be to } suspend a funnel from the fill line so the LN2 passes through the } funnel while the gas does not. This worked well in many instances. } Be sure to use a material that does not get brittle at 77 K, such as } teflon. Good luck satisfying EHS. } Yours, } Bill Tivol } 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 17 17:01:28 2003
We are in great need of a negative film scanner used to scan HRTEM films with the size of 3.25x4 inches. Although we have a Nikon super coolscan 8000 film scanner, the film holder is kind of small. It is annoying to cut the films every time when we scan them. If you use scanner to scan the electron microscopy films, please give me your suggestion for the vendor and model. Actually, this question has been discussed before, so any new comment? Thanks.
Regards, Qi Zhang
*************************************************** 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 Fax: 919-962-0480 E-mail: qizhang-at-physics.unc.edu qizhang-at-email.unc.edu
__________________________________ Do you Yahoo!? The New Yahoo! Shopping - with improved product search http://shopping.yahoo.com
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 17 17:09:41 2003
I know about the theoretical, but my question was really about 'in-practice' detection limits ie what standard deviation do you really get from replicate measurements on zero-concentration standards. The latter includes influences like beam current instability and measurement imprecision, spectrometer irregularities, etc, as well as the counting stats (as well, of course, as sample inhomogeneity).
cheers
rtch
} From: "Mary Mager" {mager-at-interchange.ubc.ca} To: "Ritchie Sims" {r.sims-at-auckland.ac.nz} Copies to: "Microscopy" {microscopy-at-MSA.microscopy.com}
If you are really that upset about cutting TEM negs for the LS 8000, well, I doubt that something else will suffice. You have the most awesome scanner available today. Cutting negs is a small chore for super high quality scans. At least I think so. Just slice them and put them in the glass carrier with 6x7 mask.
you are not going to find a 4,000dpi scanner with this kind of D range for film without going to PMT drum scanners. This can be done. but bring along about $25,000 at least. And...some require coating the negs with oil. So, would you rather soak or cut--and/or have lower resolution and lower D range?
gary g.
At 03:02 PM 10/17/2003, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 17 21:36:31 2003
Ted Clarke is requesting assistance with bloom in a Colorado lake.
"I would appreciate help from the group in identifying the species of organism causing a red film on our lake shore right after the ice melted in 1999. Our lake is considered non-polluted, but is a eutrophic lake. It has a marl bottom with a PH of 8 and a ferrous iron content of 1 ppm in the ground water. I have attached two photos showing the red film and the causing organism. The organism is unusual because it is birefringent, as shown in the images taken with crossed polars and a first order red compensator. The shots taken in the summer were from a bulk sample and no surface film was present. I think I found the same organism," Ted Clarke.
I made a full resoluion photo album at http://www.couger.com/microscope/Ted-Clarke/album/ I will make a better presentation when I am less pressed for time but the images are full resolution and all the information that I have is present. I will add more in as the discussion proceeds [gcc]
Ted can be reached buy email {tclarke-at-ligtel.com http://www.couger.com/microscope/Ted-Clarke/camera%20adapter/adapter.htm http://www.couger.com/microscope/Ted-Clarke/
Gordon Couger gcouger-at-couger.com
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.
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 18 02:14:23 2003
Hello all; The subject of scanning TEM negatives in "standard format" film scanners has once again raised its head. We bemoan the fact that TEM negatives do not conform with what scanner manufactures want to provide for the masses and slice and dice our negatives to conform. The solution lies in "older technology". For a number of years I have used a Leafscan 45 scanner to scan uncut TEM negatives with great success. This scanner is capable of scanning negatives from 35mm to 4x5 and all in.between. The Leaf uses a set of film holders that are the same as those used in Bezler sp? enlargers. To accommodate TEM negatives, I just had a machine shop modify a spare 35mm negative carrier I had to accommodate the TEM negative. Then I set the Leaf 45 to 4x5 setting and scan the whole negative. The scanner can be adjusted in software to scan the TEM negative area only. I get images of ~2500 real DPI. The file sizes are approximately 160mb. The scanner runs on both PC or Mac platforms. In Mac mode data is transferred via SCSI and in PC mode via a GPIB card. In Mac mode you can run the scanner from a Photoshop plug-in. Yay Mac. The downside to all this is that the LeafScan is no longer made but many are still available on EBAY or on the Leafscan listserver on YAHOO (egad). They generally go for about 600-1200 dollars depending on what options are offered at the time of sale. There is a chap in the Eastern US who sells refurbished scanners. I THINK his web address is www.leafstuff.com. He also carries a stock of all parts including the rather hard to find scanner lamp. The Leafscan 45 would be a much cheaper solution to scanning TEM negatives and the need for scissors would be eliminated. If anyone is interested in this solution and want more information, contact me off line.
Cheers; Jon McGovern
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 18 10:48:17 2003
I would like to purchase a negative scanner and have read the comments about the size problems with the negative carriers. I have not seen the Nikon 8000 but the specs certainly look great. Would it be feasible to have a machinist modify the existing carrier or make one that would fit the 3.25x4' film? Also has anyone approached NIKON about making a carrier for this size film?
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
On 10/17/03 6:39 PM, "Gary Gaugler" {gary-at-gaugler.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 } ---------------------------------------------------------------------------- --} - } } If you are really that upset about cutting TEM } negs for the LS 8000, well, I doubt that } something else will suffice. You have the } most awesome scanner available today. Cutting } negs is a small chore for super high quality } scans. At least I think so. Just slice them } and put them in the glass carrier with 6x7 mask. } } you are not going to find a 4,000dpi scanner with } this kind of D range for film without going } to PMT drum scanners. This can be done. but } bring along about $25,000 at least. And...some } require coating the negs with oil. So, would } you rather soak or cut--and/or have lower resolution } and lower D range? } } gary g. } } } At 03:02 PM 10/17/2003, 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 } } ----------------------------------------------------------------------------- } } -- } } } } Dear colleagues, } } } } We are in great need of a negative film scanner used } } to scan HRTEM films with the size of 3.25x4 inches. } } Although we have a Nikon super coolscan 8000 film } } scanner, the film holder is kind of small. It is } } annoying to cut the films every time when we scan } } them. If you use scanner to scan the electron } } microscopy films, please give me your suggestion for } } the vendor and model. Actually, this question has been } } discussed before, so any new comment? Thanks. } } } } Regards, } } Qi Zhang } } } } *************************************************** } } 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 } } Fax: 919-962-0480 } } E-mail: qizhang-at-physics.unc.edu } } qizhang-at-email.unc.edu } } } } } } __________________________________ } } Do you Yahoo!? } } The New Yahoo! Shopping - with improved product search } } http://shopping.yahoo.com } } } }
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 18 11:04:54 2003
Out of the box, I don't think that any modification could be made to allow the scanner to do a full TEM neg. The reason is that the film adapter units for large frames is masked off using keyed plastic masks. Each mask has a set of holes that mate with stubs sticking out of the film holder. Each mask has a unique hole patter to tell the scanner what frame size is in the holder--6x4.5cm, 6x6cm, 6x7cm. I wind up cutting the neg to fit the width of 6x7cm film and use the 6x7cm mask. This costs about 1/4" on each side of the neg, or 1/2" overall. The length does not have to be reduced.
I suspect that Nikon would have little interest in making a TEM neg holder due to low demand. If one does not have this scanner already, then probably a high end flat bed is a better solution if you don't want to cut the negs. Take a look at the Microtek 6800 (I think that is the model) which has the transparency adapter option. It is lower resolution and lower D than the Nikon 8000 but may do a satisfactory job. My UMAX Powerlook III is needing replacement and am considering the Microtek scanner--not for TEM negs though.
gary g.
At 08:48 AM 10/18/2003, you wrote: } I would like to purchase a negative scanner and have read the comments about } the size problems with the negative carriers. I have not seen the Nikon } 8000 but the specs certainly look great. Would it be feasible to have a } machinist modify the existing carrier or make one that would fit the 3.25x4' } film? Also has anyone approached NIKON about making a carrier for this size } film? } } 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 } } } } } On 10/17/03 6:39 PM, "Gary Gaugler" {gary-at-gaugler.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 } } } ---------------------------------------------------------------------------- } --} - } } } } If you are really that upset about cutting TEM } } negs for the LS 8000, well, I doubt that } } something else will suffice. You have the } } most awesome scanner available today. Cutting } } negs is a small chore for super high quality } } scans. At least I think so. Just slice them } } and put them in the glass carrier with 6x7 mask. } } } } you are not going to find a 4,000dpi scanner with } } this kind of D range for film without going } } to PMT drum scanners. This can be done. but } } bring along about $25,000 at least. And...some } } require coating the negs with oil. So, would } } you rather soak or cut--and/or have lower resolution } } and lower D range? } } } } gary g. } } } } } } At 03:02 PM 10/17/2003, 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 } } } } ----------------------------------------------------------------------------- } } } -- } } } } } } Dear colleagues, } } } } } } We are in great need of a negative film scanner used } } } to scan HRTEM films with the size of 3.25x4 inches. } } } Although we have a Nikon super coolscan 8000 film } } } scanner, the film holder is kind of small. It is } } } annoying to cut the films every time when we scan } } } them. If you use scanner to scan the electron } } } microscopy films, please give me your suggestion for } } } the vendor and model. Actually, this question has been } } } discussed before, so any new comment? Thanks. } } } } } } Regards, } } } Qi Zhang } } } } } } *************************************************** } } } 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 } } } Fax: 919-962-0480 } } } E-mail: qizhang-at-physics.unc.edu } } } qizhang-at-email.unc.edu } } } } } } } } } __________________________________ } } } Do you Yahoo!? } } } The New Yahoo! Shopping - with improved product search } } } http://shopping.yahoo.com } } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 18 11:09:36 2003
If this is for filling an EDS Dewar, check into a product called Safe Fill. It connects via a vacuum hose to a big LN2 Dewar or LN2 source and automatically fills the EDS Dewar when the level gets low. It is totally hands-off. I can get more info for you if you can't find the product.
gary g.
At 02:25 PM 10/16/2003, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 18 17:53:51 2003
There is a simple way to modify the large negative holder to get most of a TEM negative in one scan. Machine away the raised portions of the holder on the right side to accommodate the negative. You have to get pretty close to the hinge so be careful but you can get enough space to that the negative has a bit of play so there is no fear of warping. Given the time and space requirements I doubt if we will want to scan full negatives. The machining is a very simple job. Just two or three straight passes with an end mill. Regarding another thread: Image J opens the 60 meg TIF I produced without a problem. Just don't open too many at once. You can pan and scroll and play with brightness and contrast to your hearts content.
I will update you on my next project: I plan to modify the 35mm mounted slide holder to hold an EM negative in the "portrait" orientation. Then you will almost never have to do any stitching.
Michael
Gary Gaugler 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 } ------------------------------------------------------------------------------- } } } Out of the box, I don't think that any modification } could be made to allow the scanner to do a full TEM } neg. The reason is that the film adapter units for } large frames is masked off using keyed plastic masks. } Each mask has a set of holes that mate with stubs } sticking out of the film holder. Each mask has a } unique hole patter to tell the scanner what frame } size is in the holder--6x4.5cm, 6x6cm, 6x7cm. } I wind up cutting the neg to fit the width of 6x7cm } film and use the 6x7cm mask. This costs about 1/4" } on each side of the neg, or 1/2" overall. The length } does not have to be reduced. } } I suspect that Nikon would have little interest in } making a TEM neg holder due to low demand. If one } does not have this scanner already, then probably } a high end flat bed is a better solution if you } don't want to cut the negs. Take a look at the } Microtek 6800 (I think that is the model) which } has the transparency adapter option. It is lower } resolution and lower D than the Nikon 8000 but } may do a satisfactory job. My UMAX Powerlook III } is needing replacement and am considering the } Microtek scanner--not for TEM negs though. } } gary g. } } } } At 08:48 AM 10/18/2003, you wrote: } } } I would like to purchase a negative scanner and have read the } } comments about } } the size problems with the negative carriers. I have not seen the Nikon } } 8000 but the specs certainly look great. Would it be feasible to have a } } machinist modify the existing carrier or make one that would fit the } } 3.25x4' } } film? Also has anyone approached NIKON about making a carrier for } } this size } } film? } } } } 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 } } } } } } } } } } On 10/17/03 6:39 PM, "Gary Gaugler" {gary-at-gaugler.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 } } } } } ---------------------------------------------------------------------------- } } } } --} - } } } } } } If you are really that upset about cutting TEM } } } negs for the LS 8000, well, I doubt that } } } something else will suffice. You have the } } } most awesome scanner available today. Cutting } } } negs is a small chore for super high quality } } } scans. At least I think so. Just slice them } } } and put them in the glass carrier with 6x7 mask. } } } } } } you are not going to find a 4,000dpi scanner with } } } this kind of D range for film without going } } } to PMT drum scanners. This can be done. but } } } bring along about $25,000 at least. And...some } } } require coating the negs with oil. So, would } } } you rather soak or cut--and/or have lower resolution } } } and lower D range? } } } } } } gary g. } } } } } } } } } At 03:02 PM 10/17/2003, 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 } } } } } } ----------------------------------------------------------------------------- } } } } } } -- } } } } } } } } Dear colleagues, } } } } } } } } We are in great need of a negative film scanner used } } } } to scan HRTEM films with the size of 3.25x4 inches. } } } } Although we have a Nikon super coolscan 8000 film } } } } scanner, the film holder is kind of small. It is } } } } annoying to cut the films every time when we scan } } } } them. If you use scanner to scan the electron } } } } microscopy films, please give me your suggestion for } } } } the vendor and model. Actually, this question has been } } } } discussed before, so any new comment? Thanks. } } } } } } } } Regards, } } } } Qi Zhang } } } } } } } } *************************************************** } } } } 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 } } } } Fax: 919-962-0480 } } } } E-mail: qizhang-at-physics.unc.edu } } } } qizhang-at-email.unc.edu } } } } } } } } } } } } __________________________________ } } } } Do you Yahoo!? } } } } The New Yahoo! Shopping - with improved product search } } } } http://shopping.yahoo.com } } } } } } } } } } } } } } } }
-- _____________________________ W. Michael Schoel PhD Research Associate Department of Physiology & Biophysics Faculty of Medicine, University of Calgary 3330 Hospital Drive, N.W. Calgary, Alberta, Canada T2N 4N1 Phone: Office (403) 220-3662 Lab (403) 220-6674 E-mail: schoel-at-ucalgary.ca
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 20 05:33:04 2003
I have replied to a similar query back in September but will repeat an extract of the message below in case you haven't seen it
{SNIP} I have used an Epson Perfection 1200 which produce fairly good results but I was always suspicious of its quoted top resolution of 1200 dpi. Negatives on this type of flatbed scanner have to be placed on the glass surface of the scanner to be scanned which creates several problems: Newton's rings; dust and other marks on the glass.
I now use a Microtek Scanmaker 8700 which you can buy with or without Silversoft image management software (I think its the PRO version in the US). It also has a glass panel for normal flatbed document scanning but importantly has a separate glassless drawer which can take several types of film holder. See website below: http://www.microtekusa.com/sm8700.html So far I have been very impressed because the scan quality is good and some of our staff have even used the 35mm film and slide adaptors to digitise their colour slides (1200dpi is adequate for 35mm if you're using a digital projector or displaying on screen). This model comes with USB 1.1 but more usefully Firewire with an adaptor card. In the UK you also get Adobe Photoshop Elements which is quite handy. There is sufficient room to place two large 83 x 102mm (3 1/4 x 4 inch) in the glassless film carriers. Although I am still just using the 5 x 4 inch adaptors I suspect that it would be possible to construct inserts. {SNIP}
I believe Microtek do some more powerful dedicated film scanners that will take the format you desire and manage a greater optical density.
Malcolm
Malcolm Haswell e.m. unit School of Health, Natural and Social Sciences Fleming Building University of Sunderland Tyne & Wear SR1 3SD UK e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- } From: Zhang Qi {qzhangtj-at-yahoo.com}
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jcervantes-at-bendres.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, October 16, 2003 at 11:10:02 ---------------------------------------------------------------------------
Title-Subject: [Microscopy] MListserver: TEM Stain for LR-White
Question: Hello: I am working with a soft polymer dispersion, and embedding the sample in LR-White using the cold-cure method. Our sample prep is a little unconventional (I believe), as we want to avoid any alteration of the dispersion's "native" structure in order to observe evidence of phase separation, crystallization in nano-domains, etc. Thus, I do not use water, stains, etc, and microtome the blocks dry with a glass knife (what a pain), a technique used by Dr JoAn Hudson at the University of Oregon whom we have been collaborating with.
The difficulty arises in the TEM. I can not convince myself - or my colleagues - of the difference between the polymer in the dispersion and the embedding media itself (the polymer has extremely low contrast). I wondered if there was a way to label the resin (leaving the polymer alone), and came across a tip on the EMS website that suggested dying the LR-White with a fat-soluble dye (Sudan Black was listed as a good option). I wonder if anyone has experience with this sort of problem or any information on how to stain the media? I'd like to avoid the conventional techniques of staining the polymer for reasons already mentioned. Also, the second component in the dispersion is relatively electron dense and I am afraid I won't be able to tell the difference between the components in the dispersion.
Thanks in advance for any bones you can throw me!
Jessica Cervantes Bend Research, Inc Bend, OR 97701 (541) 382-0212 x240
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sergey-at-seas.ucla.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, October 20, 2003 at 11:49:51 ---------------------------------------------------------------------------
Does anybody have experience to get rid of Newton's rings on the scanned TEM images. I use Minolta Dimage Scan Multi Pro and I'm quite pleased with the quality of the scanned images except Newton's rings sometime appear on them.
Hi Gary, Thank you for the reply. I am not interested in auto fill, our LN2 vendor AirProducts actually does that as a part of their service. The downside is that we forget how much effort goes into that simple task and take it for granted. They do not issue a phase separator, but we need it to transfer LN2 for filling in cold traps.
To all of you who replied, Thank you for numerous tips.
Regards,
Jerzy
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Saturday, October 18, 2003 11:15 AM To: Gazda, Jerzy Cc: MSA listserver
If this is for filling an EDS Dewar, check into a product called Safe Fill. It connects via a vacuum hose to a big LN2 Dewar or LN2 source and automatically fills the EDS Dewar when the level gets low. It is totally hands-off. I can get more info for you if you can't find the product.
gary g.
At 02:25 PM 10/16/2003, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 20 16:10:56 2003
Looks like sort of algae. I remember Russians do find a lot of microorganisms buried very deep in pretty old Antarctic ice. Those creatures used to live in the water films on the borders between ice crystals. Some of them used to color Antarctic icebergs in funny colors. Those from the deep are anaerobic of coarse. Yours are looks like escaped ice trap and pretty happy enjoying the freedom. Sergey
At 07:36 PM 10/17/2003, 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 33-089 Los Angeles, CA 90095
Steve, I believe the way the phase separator works is this:
What is coming down the delivery tube is a boiling mix of gas and liquid at 40 to 200 psi. If it exits the open tube, there is a large amount of atomization and subsequent evaporation. In other words, most of what comes down the tube never makes it into the container. The phase separator is able to cause a lot of the fine mist to coalesce and gravity pulls the liquid to the bottom end. The gas is free to escape from the upper part, less its liquid load. The yield for the liquid part is much greater with it just running freely into the container (without pressure).
Your example shows how important it can be. I'm assuming that the valve was opened the same mount and the feed rate from the bulk tank was the same. That means that you used less than half as much from the bulk tank using the separator. And it saved you time in the bargain.
Ken Converse owner Quality Images third party SEM service Delta, PA
Stevewerner2001-at-aol.com wrote:
} Mr. Converse, } Being new to this career field (microscopy) and new to } science-as-a-livelyhood I was interested in your reasoning to the } outfitting of the live end of the LN line. I have experienced the same } thing here at my microscopy job: pre-frit it took about 42 minutes to } fill the first empty 35 liter dewar on a 85-87 degree day at (usual?) } pressure. Now with the frit nozzle an mere 18 minutes. The distance to } the bulk tank has not changed. Why does a nozzle, that appeares to be } an arreator, cause the fluid to remain fluid and not phase into a gas? } } I understand that you are a business operater and if you don't have } time or reason to respond, that is OK. Any reply is well appreciated. } Thank you, Steve Werner.
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 21 07:48:16 2003
I have been told by knowledgable folks that coating the (face down side of the) negative with "Kami mounting fluid" will stop the formation of Newton rings. Haven't tried it.
Geoff
by way of MicroscopyListserver wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- -- ********************************************** 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 Tue Oct 21 08:27:03 2003
My colleague is using fluorescent-labeled antibodies to stain bacteria living on agar plates. He is currently imaging these stains/bacteria by picking up the bacteria and placing them on slides. He says that he often is not getting the best images, most likely from interference and background from the agar. Is there a better way to isolate the bacteria from the agar? Should he use a water immersion objective? I've never worked with bacteria before, so I'm finding it a bit difficult to help him in this situation.
Thanks so much,
Elke Pravda Biostructure Core Facility Forsyth Institute Boston, MA epravda-at-forsyth.org
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 21 11:05:54 2003
If one is using an LN2 tank at 40 to 200 psi that is a major part of the liquid delivery problem. A tank intended to deliver liquid, as opposed to gas-phase coolant, should be a low pressure tank that maintains the liquid only under about 20 psi, enough to boost it out the nozzle and little more. Most delivery services can provide either low pressure (~20 psi) tanks or high pressure tanks. The typical 110 L size of each is very similar in appearance except for the plumbing and pressure regulation apparatus at the top. The high pressure tanks are preferred by users of temperature cycling equipment where the objective is to maximize cooling throughout a protracted discharge from the tank without delivering liquid into the test chamber. It can be virtually impossible to collect the liquid effectively from a high pressure tank.
John Twilley
qualityimages 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 } ------------------------------------------------------------------------------- } } Steve, } I believe the way the phase separator works is this: } } What is coming down the delivery tube is a boiling mix of gas and liquid } at 40 to 200 psi. If it exits the open tube, there is a large amount of } atomization and subsequent evaporation. In other words, most of what } comes down the tube never makes it into the container. The phase } separator is able to cause a lot of the fine mist to coalesce and } gravity pulls the liquid to the bottom end. The gas is free to escape } from the upper part, less its liquid load. The yield for the liquid } part is much greater with it just running freely into the container } (without pressure). } } Your example shows how important it can be. I'm assuming that the valve } was opened the same mount and the feed rate from the bulk tank was the } same. That means that you used less than half as much from the bulk } tank using the separator. And it saved you time in the bargain. } } Ken Converse } owner } Quality Images } third party SEM service } Delta, PA } } Stevewerner2001-at-aol.com wrote: } } } Mr. Converse, } } Being new to this career field (microscopy) and new to } } science-as-a-livelyhood I was interested in your reasoning to the } } outfitting of the live end of the LN line. I have experienced the same } } thing here at my microscopy job: pre-frit it took about 42 minutes to } } fill the first empty 35 liter dewar on a 85-87 degree day at (usual?) } } pressure. Now with the frit nozzle an mere 18 minutes. The distance to } } the bulk tank has not changed. Why does a nozzle, that appeares to be } } an arreator, cause the fluid to remain fluid and not phase into a gas? } } } } I understand that you are a business operater and if you don't have } } time or reason to respond, that is OK. Any reply is well appreciated. } } Thank you, Steve Werner.
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 21 14:09:31 2003
I've dug through the documentation I've got but haven't found anything.
Does anyone, have a suggestion for getting it clean inside?
Most of the use now is with ethanol suspended copper nano-particles. And there has been a bit of a build up/discoloration of some of the parts. And with a bit of glass this delicate and expensive I just want to be extra cautious.
Thanks,
Geoff Williams Microscopy Facility Supervisor
CMU Biology Department Microscopy Facility web page. http://www.cst.cmich.edu/centers/microscopy/
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 21 14:53:43 2003
We have a JEOL 1200EX that uses freon in the HV tank.
Yesterday a campus enforcer raided the lab and confiscated our supply of Freon 12 used to replenish the HV tank and just about took me to jail for having it.
I understand and sympathize with the efforts to contain this stuff, but you would think it was heroin or enriched uranium the way this guy acted. He wants it inventoried, under lock and key, and will keep it 'for safe keeping' at the central campus facilities site. If we need it, we can call him and he will arrange for a technician to come to the lab and maybe top off the tank.
No amount of explaining that we hardly ever use it or that when we need to use it we are careful or that my independent service provider doesn't carry this stuff around in his car would satisfy him. We don't have any gross leaks and have only kept the tank around because it came with the microscope and there are directions for checking the HV tank etc. You know, just in case we need it and it is a part of the instrument.
Has anyone had a similar problem? How did you resolve it? Are there any alternatives to this scenario?
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 Tue Oct 21 14:57:14 2003
A few decades ago I made many 3.25x4 inch lantern slides (pre-35mm slide presentations). To elimintate Newton Ring formation between the glass slide and the glass cover, we put a frame of paper between them. I do not know the debth of the focus plane in your scanner but if it is not necessary to have the negative in direct contact with the scanner plate, you can easily cut a rectangle out of a piece of paper or thin cardboard, slightly smaller than your negative, put the negative on top and try it out. Perhaps a top frame is also be needed. If this is true a file folder can be used. Inexpensive and not at all messy.
The scanner that I currently use can accomodate a plastic negative holder that was made for it. It must be at least 1mm from the glass plate to the negative surface and the image is still in focus.
Pat Stranen Connelly The University of Pennsylvania Department of Biology Philadelphia, PA 19104-6018 215-898-7145 psconnel-at-sas.upenn.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 21 15:22:02 2003
Dear Michael, I don't know if anyone answered your questions, so here is what I know. 1. the Ovonics company originally marketed the layered pseudocrystals with the big d-spacing for the efficient diffraction of light-element x-rays. A search of the Web found an Ovonics company, but I don't know if they are the same. You might contact Oxford Instruments, particularly Joe Carr, their WDX specialist, to see if they are still around, because everyone uses these crystals for light elements. 2. My P-10 gas is delivered with a two-stage regulator set for ten PSI. The original tank I got when the spectrometer was installed in 1987 is still going, over half full. Regards, Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "Michael Cheatham" {mmcheath-at-mailbox.syr.edu} To: {microscopy-at-ns.microscopy.com} Sent: Thursday, October 16, 2003 10:02 AM
A few years ago we had the tank of a JEOL 2000 converted from Freon to SF6 for the same reason. I don't recall the cost but it wasn't way over the top. I will check. Perhaps you will need to do this, too.
cheers, John
******** John S. Vetrano Sr. Research Scientist Materials Structure and Performance Group Pacific Northwest National Laboratory MSIN P8-16 P.O. Box 999 Richland, WA 99352 Phone: (509)372-0724 Fax: (509)376-6308 Email: mailto:john.vetrano-at-pnl.gov
} ---------- } From: Jon Krupp } Sent: Tuesday, October 21, 2003 12:53 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: [Microscopy] Freon alternatives? } } } } ------------------------------------------------------------------------------ } 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 have a JEOL 1200EX that uses freon in the HV tank. } } Yesterday a campus enforcer raided the lab and confiscated our supply of } Freon 12 used to replenish the HV tank and just about took me to jail for } having it. } } I understand and sympathize with the efforts to contain this stuff, but you } would think it was heroin or enriched uranium the way this guy acted. He } wants it inventoried, under lock and key, and will keep it 'for safe } keeping' at the central campus facilities site. If we need it, we can call } him and he will arrange for a technician to come to the lab and maybe top } off the tank. } } No amount of explaining that we hardly ever use it or that when we need to } use it we are careful or that my independent service provider doesn't carry } this stuff around in his car would satisfy him. We don't have any gross } leaks and have only kept the tank around because it came with the } microscope and there are directions for checking the HV tank etc. You know, } just in case we need it and it is a part of the instrument. } } Has anyone had a similar problem? How did you resolve it? Are there any } alternatives to this scenario? } } 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 Tue Oct 21 20:54:59 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (martin.hohmann-marriott-at-asu.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, October 21, 2003 at 18:17:41 ---------------------------------------------------------------------------
Email: martin.hohmann-marriott-at-asu.edu Name: Martin Hohmann-Marriott
Organization: Arizona State University
Title-Subject: [Microscopy] Fixation & embedding with DMSO
Question: Dear hard-working microscopists,
Unfortunatelly, alcohols and acetone dissolve a biological structure that I am interested in visualizing. Therefore I am looking for alternative solvents.
-Is it possible to fix cells in DMSO? -Does DMSO interacct with OsO4? -Does it interact with glutaraldehyde? -Is there a resin that mixes with DMSO?
Jean-Baptiste Comiti has a web page in French on scanning light photography using equipment he built. The only thing more stunning than his images is the equipment he built to get them.
Babel Fish does a passable translation http://babelfish.altavista.com/babelfish/tr enter http://comiti.com in translate a web page and chose French to English. All computer translations have some awkward phasing and part that incorrect but you should be able to follow this one OK.
Thanks to Ted Clarke for bringing this to my attention.
Gordon Couger gcouger-at-couger.com
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.
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 22 01:52:26 2003
1) Replace R-12 with SF6, if this is OK with JEOL.
2) Have any refrigeration service (large one or a single contractor) to store your R-12 for you, and refill HT tank when necessary. They charge from $30 to $80 per hour.
3) Get EPA license for handling refrigerants yourself. It did cost me half-a-day and $80 to take a test. Test is easy. Such license allows purchasing, handling, storing, and using refrigerants, including severely restricted chlorocarbons such as R-12 and others, in all USA. You will be required to have proper equipment, which is simple and inexpensive for refills (refrigeration service valves and hoses). Main rule- do not vent R-12 on purpose. Leaks are OK- same as with old household refrigerator or AC (air conditioner). Just refill alone is easy. But, ask your AC service people to pump out freon when and if HT tank needs repair- this will require more expensive equipment and more work. Make sure that your organization is still part of USA and does not have different rules. I am serious :-)
More important point is that R-12 is not manufactured for a number of years. You can still buy it (recycled), but last time I checked it was close to $1,100 per pound (eleven hundred). Perhaps SF6 is the best way to go.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax (678)467-0012 mobile www.sia-cam.com
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----- Original Message ----- } From: "Jon Krupp" {jmkrupp-at-cats.ucsc.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, October 21, 2003 3:53 PM
Jonathan,
I work on automotive air conditioning as a hobby. Perhaps the easiest way to resolve this problem is as Vitaly suggested, to certify one of the cognizant personnel in the lab for use of r12. Though I haven't certified myself, I heard it's quite quick and painless to do online and costs only $20. Click on (http://www.epatest.com/) to get started. Once you have your license, you're certified to own and use r12 refrigerant. This will take the teeth out of the enforcers. For actual service that involves removing refrigerant, you may want to contract a refrigeration specialist.
I heard the price of r12 has actually come down to $20 per 12 oz. can. This is because a lot of pre-1995 cars that use r12 are being retired from service. The used freon supply is increasing and demand for use is decreasing. Freon is available on eBay (with proof of a license).
Be careful with conversions. I'm not familiar with SF6 refrigerant. But if you convert, make sure the lubricant is compatible. If it isn't, the entire refrigeration system will need to be flushed with solvent during the conversion and recharged with the proper type and amount of lubricant. Your cheapest and least compromising option is probably to stay with r12 in your refrigerator.
A good source of information on refrigeration can be found on: http://www.aircondition.com/wwwboard/
Stu Smalinskas Senior Metallurgist SKF North American Technical Center Plymouth, Michigan (734) 414-6862 stu.smalinskas-at-skf.com
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Jonathan wrote:
We have a JEOL 1200EX that uses freon in the HV tank.
Yesterday a campus enforcer raided the lab and confiscated our supply of Freon 12 used to replenish the HV tank and just about took me to jail for having it.
I understand and sympathize with the efforts to contain this stuff, but you would think it was heroin or enriched uranium the way this guy acted. He wants it inventoried, under lock and key, and will keep it 'for safe keeping' at the central campus facilities site. If we need it, we can call him and he will arrange for a technician to come to the lab and maybe top off the tank.
No amount of explaining that we hardly ever use it or that when we need to use it we are careful or that my independent service provider doesn't carry this stuff around in his car would satisfy him. We don't have any gross leaks and have only kept the tank around because it came with the microscope and there are directions for checking the HV tank etc. You know, just in case we need it and it is a part of the instrument.
Has anyone had a similar problem? How did you resolve it? Are there any alternatives to this scenario?
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
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From MicroscopyL-request-at-ns.microscopy.com Wed Oct 22 08:36:08 2003
I was going to suggest "chemical" dehydration with 2,2 dimethoxypropane but then I found out that it yeilds methanol and acetone which you want to avoid. How about dioxane? Or could you just skip solvents and dehydrate by critical point drying or freeze drying, then embed in whatever resin you choose?
Geoff
by way of MicroscopyListServer 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 (martin.hohmann-marriott-at-asu.edu) from } http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on } Tuesday, October 21, 2003 at 18:17:41 } --------------------------------------------------------------------------- } } } Email: martin.hohmann-marriott-at-asu.edu } Name: Martin Hohmann-Marriott } } Organization: Arizona State University } } Title-Subject: [Microscopy] Fixation & embedding with DMSO } } Question: Dear hard-working microscopists, } } Unfortunatelly, alcohols and acetone dissolve a biological structure } that I am interested in visualizing. Therefore I am looking for } alternative solvents. } } -Is it possible to fix cells in DMSO? } -Does DMSO interacct with OsO4? } -Does it interact with glutaraldehyde? } -Is there a resin that mixes with DMSO? } } Thank you for your help } Cheers, } } Martin Hohmann-Marriott } } --------------------------------------------------------------------------- } }
-- -- ********************************************** 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 Wed Oct 22 10:16:13 2003
Recently I have been asked to run EDS on scraps and chips of steel alloys. The surfaces are too rough and irregular for quantification but I would like to run standards of different stainless steels to get a ball park idea of relative element responce. Is anyone aware of a source of inexpensive steel reference standards. I don't need NIST traceablity, I just need to know the ASTM alloy type (316 or what ever).
Thanks!!
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 22 11:17:19 2003
Freon is used in the tank as an insulator not as a refrigerant.
SF6 is the only (costly) alternative. On a JEOL 1200EX it is not just a case of removing the Freon and replacing it with SF6. It is necessary to get JEOL in to replace components in the HT tank in order to make it compatible with SF6.
Regards
Alan
At 06:21 AM 10/22/2003 -0700, Kestutis Smalinskas wrote:
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Alan W Nicholls, PhD Director of Research Service Facility (Electron Microscopy) Research Resources Center - East (M/C 337) Room 100 Science and Engineering South Building The University of Illinois at Chicago 845 West Taylor St Chicago, IL 60607-7058
Please contact me off line if you are interested in a 1980s vintage Philips 430 TEM with one single tilt holder. The price has been reduced considerably. The buyer must pay shipping and handling charges. Full payment is required prior to shipping from ASU.
Please contact me today (I won't be here Thursday and Friday) or next week by e-mail or phone.
If you have specific questions regarding payment, please call Patricia Chase, Surplus Property Manager at 480-965-1850 or e-mail: patricia.chase-at-asu.edu.
John C. Wheatley Lab Manager Arizona State University Center for Solid State Science PSA-213 BOX 871704 Tempe, AZ 85287-1704
Any steel supplied by a reputable metal supply company will come with a certified mill test report (CMTR) containing composition data. You might be able to work out a deal with a friendly neighborhood steel supplier to obtain small samples of several different bars and alloys with a CMTR for each little sample. They might be able to provide you with short remnants from larger bars, or if they are really friendly, let you cut off a small piece that would not diminish the value of a full bar. This could allow you to get several "standards" at minimal cost.
-- Larry D. Hanke, P.E. Materials Evaluation and Engineering, Inc. Practical Solutions Through Technology and Innovation http://www.mee-inc.com (763) 449-8870
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 22 13:14:24 2003
You should be able to get a pretty good idea of the composition even without flat pieces and probably without SS references. We have one client that is sending us samples in almost every form: chucks, pieces, shavings, drillings, scrapings, etc. They are trying to get an idea of what materials are present throughout their food processing plant so that they can narrow down the source should a piece find its way into their food stream. We find that they have a lot of stainless steel of about 18% Cr content. Sometimes we find a piece with 20% Cr, but I am reluctant to get too excited about the change in content. It might well be a real difference, but I am not sure what tolerances are allowed on alloy compositions.
Since most of the alloying elements are at moderate energy 5-10 kV, the intensities are not terribly affected by geometry. Still, I try to find a flat and level place on the sample to minimize the effects. If elements are being determined from lower energy lines (1-3 kV), then I have to be somewhat suspicious of the results. Intensities can vary significantly based on the local geometry. Fortunately, many alloys are not specified too tightly, e.g., 0.4-0.8% of an element. It may be enough to determine the presence of an element.
You can try some experiments to determine the sensitivity to geometry. You might put a rod in your SEM perpendicular to your x-ray detector axis and move the location of the analysis to see how the results change with tilt. For that matter, if you can tilt your stage both ways with respect to the detector, you could perform the same exercise with a polished, flat sample and know the geometry exactly. You might also try some shavings with roughness and change the size of the raster used to scan the sample and see what effect that has on the results.
As you develop familiarity with what a "normal" spectrum and background looks like (e.g., position of the background hump) you should be able to recognize what should be good spectra from your samples. That should help improve your results. Of course, flattening out the samples as well as you can before analysis will also help.
Warren
At 11:02 AM 10/22/2003 -0400, you wrote:
} Recently I have been asked to run EDS on scraps and chips of steel alloys. } The surfaces are too rough and irregular for quantification but I would } like to run standards of different stainless steels to get a ball park idea } of relative element responce. Is anyone aware of a source of inexpensive } steel reference standards. I don't need NIST traceablity, I just need to } know the ASTM alloy type (316 or what ever). } } Thanks!! } } Frank Karl } Degussa Corporation } Akron Technical Center } 3500 Embassy Parkway } Suite 100 } Akron, Ohio 44333 } } } 330-668-2235 Ext. 238
------------------------------------------- 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
} Unfortunatelly, alcohols and acetone dissolve a biological structure that } I am interested in visualizing. Therefore I am looking for alternative } solvents. } } -Is it possible to fix cells in DMSO?=====} DMSO will not chemically fix } -Does DMSO interact with OsO4?=====} perhaps no, check Merk Index for } compatibility. } -Does it interact with glutaraldehyde?======} see above } -Is there a resin that mixes with DMSO?====} perhaps no
Dear Martin I don't understand your point. How do you know that alcohol and acetone dissolve your structure? Did you chemically fix your sample before? If so, it quite unlikely that your "structure" will be "dissolved". I am surprise: DMSO is such good solvent, it should "dissolve" even better than ethanol or acetone. Nevertheless, you may try some water-compatible embedding media to avoid organics. Some acrylates may do the job. Durcupan also comes to my mind (old stuff). Another choice is to use "freeze-substitution" - it's shown that at low temperature acetone has much smaller "dissolving" ability... Good luck, Sergey
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 22 14:28:35 2003
I installed digital locks on all my Lab doors, so nobody without PIN could enter (including the head of Department). The idea is that EM "presence" risk to your health, so you have to be trained and certified (by me) in order to have access to that "hazardous, nasty" place... So, everyone who want to enter my Lab should be "certified" by me. To made it more "professional" they actually have to sign a special form... I also established the strict rule that everyone who want to see my rooms should set appointment with me. My previous experience with Russian bureaucracy helps me so much here in US. The bottom line here - to do everything legally and in agreement with local laws. Local policies are not a law by the way. Sergey
At 06:21 AM 10/22/2003, you wrote:
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_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
Thanks for your comments and suggestion for the negative film scanner. By comparing the specifications and prices of scanners your suggested, we decided to modify our present scanner holder. The holder type we are using for TEM films is Nikon FH-869S. We polished the two strips, which were used to fix the film size. The two strips are much easy to be milled by lathe in our machine shop. Then we got the space, which is a little bit larger than the film size. Right now, the films are fixed just by two sides’ upper holders with 4 hooks, however, the films are as stable as before. If anyone who is using this kind of holder has the similar problem as we have, please try this way to modify your holder. Thanks all again.
Regards, Qi Zhang
*************************************************** 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 Fax: 919-962-0480 E-mail: qizhang-at-physics.unc.edu qizhang-at-email.unc.edu
--- Zhang Qi {qzhangtj-at-yahoo.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 } ------------------------------------------------------------------------------- } } Dear colleagues, } } We are in great need of a negative film scanner used } to scan HRTEM films with the size of 3.25x4 inches. } Although we have a Nikon super coolscan 8000 film } scanner, the film holder is kind of small. It is } annoying to cut the films every time when we scan } them. If you use scanner to scan the electron } microscopy films, please give me your suggestion for } the vendor and model. Actually, this question has } been } discussed before, so any new comment? Thanks. } } Regards, } Qi Zhang } } *************************************************** } 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 } Fax: 919-962-0480 } E-mail: qizhang-at-physics.unc.edu } qizhang-at-email.unc.edu } } } __________________________________ } Do you Yahoo!? } The New Yahoo! Shopping - with improved product } search } http://shopping.yahoo.com }
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From MicroscopyL-request-at-ns.microscopy.com Wed Oct 22 15:09:22 2003
Dear Frank, When I am trying to identify alloys, I use the listings in the American Society of Metals Handbook. They list the composition ranges for most of the alloys known. There is a desk addition (the full Handbook is 16 or more volumes) that covers the basics. You might go to the ASM web site and see if they sell a book or CD that will give you the basic alloy compositions. Regards, Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: {Frank.Karl-at-degussa.com} To: {microscopy-at-msa.microscopy.com} Sent: Wednesday, October 22, 2003 8:02 AM
I wonder if the new replacement for R-12 will work ( I think it is 134). It is compatible with the same seals in refrigeration systems. The only problem is compatibility with the lubricants, but that is not a problem here. When I needed to stop using R-114, I called DuPont and spoke with an engineer. I explained what my system was doing with the R-114, and he helped me find an alternative in the new "SUVA" refrigerants. The specification of interest here would be the dielectric properties and the composition of the seals. There are still rules on the handling of it, but it is not looked at as if it is going cause the end of the universe, which would, of course, find us all at Milliways.
Sorry, I strayed a bit. If it were me, I would call DuPont and talk to an engineer there. If you switch to something not on the EPA hit list, the campus enforcers should be more willing to work with you. You will, of course need a qualified contractor to remove the R-12 in an approved manner. You may be allowed to do the top-offs yourself, afterwards. They sell the new refrigerant in auto parts stores for home use.
Regards, Darrell
Alan Nicholls {nicholls-at-uic.edu To: microscopy-at-sparc5.microscopy.com } cc: Subject: [Microscopy] Re: Re: Freon alternatives? 10/22/2003 12:25 PM
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Jonathon
Freon is used in the tank as an insulator not as a refrigerant.
SF6 is the only (costly) alternative. On a JEOL 1200EX it is not just a case of removing the Freon and replacing it with SF6. It is necessary to get JEOL in to replace components in the HT tank in order to make it compatible with SF6.
Regards
Alan
At 06:21 AM 10/22/2003 -0700, Kestutis Smalinskas wrote:
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } Jonathan, } } I work on automotive air conditioning as a hobby. } Perhaps the easiest way to resolve this problem is as } Vitaly suggested, to certify one of the cognizant } personnel in the lab for use of r12. Though I haven't } certified myself, I heard it's quite quick and } painless to do online and costs only $20. Click on } (http://www.epatest.com/) to get started. Once you } have your license, you're certified to own and use r12 } refrigerant. This will take the teeth out of the } enforcers. For actual service that involves removing } refrigerant, you may want to contract a refrigeration } specialist. } } I heard the price of r12 has actually come down to $20 } per 12 oz. can. This is because a lot of pre-1995 } cars that use r12 are being retired from service. The } used freon supply is increasing and demand for use is } decreasing. Freon is available on eBay (with proof of } a license). } } Be careful with conversions. I'm not familiar with } SF6 refrigerant. But if you convert, make sure the } lubricant is compatible. If it isn't, the entire } refrigeration system will need to be flushed with } solvent during the conversion and recharged with the } proper type and amount of lubricant. Your cheapest } and least compromising option is probably to stay with } r12 in your refrigerator. } } A good source of information on refrigeration can be } found on: http://www.aircondition.com/wwwboard/ } } Stu Smalinskas } Senior Metallurgist } SKF North American Technical Center } Plymouth, Michigan } (734) 414-6862 } stu.smalinskas-at-skf.com } } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } Jonathan wrote: } } We have a JEOL 1200EX that uses freon in the HV tank. } } Yesterday a campus enforcer raided the lab and } confiscated our supply of } Freon 12 used to replenish the HV tank and just about } took me to jail for } having it. } } I understand and sympathize with the efforts to } contain this stuff, but you } would think it was heroin or enriched uranium the way } this guy acted. He } wants it inventoried, under lock and key, and will } keep it 'for safe } keeping' at the central campus facilities site. If we } need it, we can call } him and he will arrange for a technician to come to } the lab and maybe top } off the tank. } } No amount of explaining that we hardly ever use it or } that when we need to } use it we are careful or that my independent service } provider doesn't carry } this stuff around in his car would satisfy him. We } don't have any gross } leaks and have only kept the tank around because it } came with the } microscope and there are directions for checking the } HV tank etc. You know, } just in case we need it and it is a part of the } instrument. } } Has anyone had a similar problem? How did you resolve } it? Are there any } alternatives to this scenario? } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } } __________________________________ } Do you Yahoo!? } The New Yahoo! Shopping - with improved product search } http://shopping.yahoo.com
Alan W Nicholls, PhD Director of Research Service Facility (Electron Microscopy) Research Resources Center - East (M/C 337) Room 100 Science and Engineering South Building The University of Illinois at Chicago 845 West Taylor St Chicago, IL 60607-7058
Robert Fowler Quality Assurance Failure Analysis Technician TDK Components USA, Inc. Multilayer Ceramic Capacitor Division 1 TDK Boulevard Peachtree City GA 30269-2051 Telephone: (770) 631-0410 Ext.315 Fax: (770) 487-1460 email: rfowler-at-tdktca.com www.component.tdk.com ----- Forwarded by Robert Fowler/TCU/TDK-US on 10/23/2003 07:44 AM -----
Robert Fowler To: Microscopy-at-sparc5.microscopy.com 10/22/2003 cc: 02:01 PM Subject: [Microscopy] Help with Nikon 990 Threaded Adapter
Listers My Nikon 990 threaded adapter (on the camera end) has seen its better days after repeated threading and unthreading (I know shame on me for not dedicating....budget). It now has no more threads to attach to the MDC/c-mount adapter. Does anyone out there have a solution to this problem? Is there a threaded adapter available from Nikon or elsewhere? Any solution is better than no solution. Thank you
BTW Sorry if this is double posted had some trouble with my subscription
Also I talked to many people at Nikon before someone informed me this was not a replacement part so ALL comments are welcome
Robert Fowler Quality Assurance Failure Analysis Technician TDK Components USA, Inc. Multilayer Ceramic Capacitor Division 1 TDK Boulevard Peachtree City GA 30269-2051 Telephone: (770) 631-0410 Ext.315 Fax: (770) 487-1460 email: rfowler-at-tdktca.com www.component.tdk.com
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From MicroscopyL-request-at-ns.microscopy.com Thu Oct 23 07:48:17 2003
DMSO isn't a fixative, but can be added to improve fixative penetration. It doesn't react with glutaraldehyde and is commonly used in combination with it. Use caution with this combination so as to not fix yourself, too. It is my understanding that DMSO does, however, react badly with osmium tetroxide forming a tar-like substance in and around your samples.
Robert Simmons
Dr Robert Simmons Program Director Biological Imaging Core Laboratory Georgia State University Atlanta, GA 30302-4010
404-651-3138 404-651-2509 FAX
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 23 13:30:37 2003
My supervisor has just requested our travel and training needs for the upcoming year. I am new to TEM and would like to attend some biological type seminars. Does anyone have any suggestions or recommendations for future meetings?
-- Sue Tyler Biologist Cooperative Oxford Laboratory Center for Coastal Environmental Health Biomolecular Research at Charleston ( CCHEBR) USDOC/NOAA/NOS/NCCOS 904 S. Morris St. Oxford, Maryland 21654-9724 410-226-5193 Fax: 410- 226-5925 Sue.Tyler-at-noaa.gov
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 23 14:18:14 2003
Dear Martin How do you know DMSO is safe? Also, if your structures are soluble in alcohol and acetone, there is a good chance they will be dissolved or damaged by the resins themselves at 20oC unless they are chemically fixed. Sergey is right - you should consider using freeze-substitution and low temperatures for the entire process - dehydration, infiltration with resin and polymerization. Power of solvents is dramatically altered by temperature. e.g. fats and waxes soluble in chloroform at +20oC become insoluble at -20oC.
But none of us can give you really good advice unless you tell us what your structure is, or at least its composition.
Chris
Dr. Chris Jeffree
} ----- Original Message ----- } From: "Sergey Ryazantsev" {sryazant-at-ucla.edu} } To: {Microscopy-at-sparc5.microscopy.com} } Sent: Wednesday, October 22, 2003 8:10 PM } Subject: [Microscopy] Re: via-WWW: Fixation & embedding with DMSO } } } } } } } } -------------------------------------------------------------------- } ---------- } } 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 } } -------------------------------------------------------------------- } ----------- } } } } } } } Unfortunatelly, alcohols and acetone dissolve a biological } structure that } } } I am interested in visualizing. Therefore I am looking for } alternative } } } solvents. } } } } } } -Is it possible to fix cells in DMSO?=====} DMSO will not } chemically fix } } } -Does DMSO interact with OsO4?=====} perhaps no, check Merk Index } for } } } compatibility. } } } -Does it interact with glutaraldehyde?======} see above } } } -Is there a resin that mixes with DMSO?====} perhaps no } } } } Dear Martin } } I don't understand your point. How do you know that alcohol and } acetone } } dissolve your structure? Did you chemically fix your sample before? } If } } so, it quite unlikely that your "structure" will be "dissolved". I } am } } surprise: DMSO is such good solvent, it should "dissolve" even } better than } } ethanol or acetone. Nevertheless, you may try some water-compatible } } embedding media to avoid organics. Some acrylates may do the job. } } Durcupan also comes to my mind (old stuff). Another choice is } to use } } "freeze-substitution" - it's shown that at low temperature acetone } has much } } smaller "dissolving" ability... Good luck, Sergey } } } } }
------- End of forwarded message ------- ========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 (0) 131 650 5554 FAX. #44 (0) 131 650 5392 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 23 14:19:58 2003
I don't know if our adapter would help you (it is a 28mm threaded adapter as well). But, if you would like to order one and give it a shot, you can do so with a 30 day no-hassle return policy.
What makes our adapters better? http://www.mvia.com/Coolpix/clpxadpt.htm#What_makes_Our_Adapters_Better
Thanks! Jim Haley
****************************** Jim Haley Applications Engineer MVIA, Inc. 125 Sherwood Drive Monaca, PA 15061 voice: (724) 728-7493 fax: (412) 291-1709 e-mail: haley-at-mvia.com webpage: http://www.mvia.com ******************************
Robert.Fowler-at-tdktca.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 } ------------------------------------------------------------------------------- } } Robert Fowler } Quality Assurance Failure Analysis Technician } TDK Components USA, Inc. } Multilayer Ceramic Capacitor Division } 1 TDK Boulevard } Peachtree City GA 30269-2051 } Telephone: (770) 631-0410 Ext.315 } Fax: (770) 487-1460 } email: rfowler-at-tdktca.com } www.component.tdk.com } ----- Forwarded by Robert Fowler/TCU/TDK-US on 10/23/2003 07:44 AM ----- } } Robert Fowler } To: Microscopy-at-sparc5.microscopy.com } 10/22/2003 cc: } 02:01 PM Subject: [Microscopy] Help with Nikon 990 Threaded Adapter } } } } Listers } My Nikon 990 threaded adapter (on the camera end) has seen its better days } after repeated threading and unthreading (I know shame on me for not } dedicating....budget). It now has no more threads to attach to the } MDC/c-mount adapter. Does anyone out there have a solution to this problem? } Is there a threaded adapter available from Nikon or elsewhere? Any } solution is better than no solution. Thank you } } BTW Sorry if this is double posted had some trouble with my subscription } } Also I talked to many people at Nikon before someone informed me this was } not a replacement part so ALL comments are welcome } } Robert Fowler } Quality Assurance Failure Analysis Technician } TDK Components USA, Inc. } Multilayer Ceramic Capacitor Division } 1 TDK Boulevard } Peachtree City GA 30269-2051 } Telephone: (770) 631-0410 Ext.315 } Fax: (770) 487-1460 } email: rfowler-at-tdktca.com } www.component.tdk.com } } THIS TRANSMISSION IS INTENDED FOR THE SOLE USE OF THE INDIVIDUAL AND ENTITY } TO WHOM IT IS ADDRESSED AND MAY CONTAIN PRIVILEGED AND/OR CONFIDENTIAL } INFORMATION. } } If you are not the intended recipient, be advised that any use, } dissemination, distribution or duplication of this transmission is strictly } prohibited. If you received this transmission in error, please notify the } sender immediately by electronic reply to this transmission or by phone } (847-803-6100). Thank you.
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From MicroscopyL-request-at-ns.microscopy.com Thu Oct 23 17:22:02 2003
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax (678)467-0012 mobile www.sia-cam.com
This message is made of 100% recycled electrons.
This address can not receive messages larger than 15 kb without prior notification.
----- Original Message ----- } From: {Robert.Fowler-at-tdktca.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, October 23, 2003 7:44 AM
Thanks for the response.
the info I seek is specific to STEM on/in an SEM. Major SEM makers offer a STEM option. Some tough the BF and DF feature. I just don't understand how a dark field STEM image is acquired on a SEM which has a STEM stage option.
gary g.
At 06:56 PM 10/23/2003, you wrote: } Gary, } } The detectors are the same, the difference is in geometry. Bright field is } a circle, and dark field is a ring around the circle. In } a properly aligned STEM all that is required to switch from BF to DF is to } switch the sensors. Both BF and DF signals are present } all time, it matters what sensor you are acquiring from. I am not sure } what you mean by SEM. This sensor arrangement will produce } results identical for both sensors in SEM. Did you mean TEM? Sensors, BTW, } are same (well, almost) as for SEM backscattered } electrons detectors. I can look up something and fax it to you if you } wish. Several pages. } } Vitaly Feingold } Scientific Instruments and Applications } 2773 Heath Lane, Duluth GA 30096 } (770)232-7785 ph. } (770)232-1791 fax } (678)467-0012 mobile } www.sia-cam.com } } This message is made of 100% recycled electrons. } } This address can not receive messages larger than 15 kb without prior } notification. } } ----- Original Message ----- } From: "Gary Gaugler" {gary-at-gaugler.com} } To: "MSA listserver" {Microscopy-at-MSA.Microscopy.Com} } Sent: Thursday, October 23, 2003 6:31 PM } Subject: [Microscopy] Darkfield STEM } } } } } } } } } ------------------------------------------------------------------------------ } } 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: } } } } Is there a reference source that explains how } } a BF/DF STEM detector works on a SEM? In } } particular, how does it do DF? } } } } tnx, } } gary g. } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 24 02:25:35 2003
http://www.microscopy-uk.net/mag/artaug01/vrcoolpix.htm and http://www.microscopy-uk.net/mag/artsep01/vrcoolpix2.htm
It's a way to mount a coolpix or any other digital camera on a microscope, using the tripod thread insted of the objectif thread. You'll need perheps to modify your microscope interface. Of coarse you need a workshop ! It works well, particulary if you can have a centring of your interface inside (or outside) the "late thred" of the camera objectif.
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
Sorry folks, but I can't seem to post the list these days, please respond directly to me: Richard Edelmann {edelmare-at-MUOHIO.EDU} and not the sender of this message
------- Forwarded message follows ------- } From: Richard Edelmann {edelmare-at-MUOHIO.EDU} To: emsabbs
Gary
The geometry, what is and how it is collected is exactly the same as STEM in TEM.
The BF/DF detectors are after the specimen and collect electrons that have been transmitted through the (thin) specimen. The BF detector collects electrons close to the optical axis which have not suffered a scattering interaction with the specimen (direct beam). The Annular DF detector, which is a ring scintillator with a hole in the center, simultaneously collects electrons that have been forward scattered from the specimen in to a particular angular range (typical 10mrad to maybe 300mrad). Close to the direct beam these are diffracted and inelastically scattered electrons, further out these are elastically scattered electrons. Assuming the dark field image collects most of the scattered electrons the image will look like the negative of the bright field image.
The technique is briefly mentioned in "Scanning Electron Microscopy and X-ray Microanalysis" by Goldstein et al. and also more extensively in "Transmission Electron Microscopy" by Williams and Carter
Alan
At 07:10 PM 10/23/2003 -0700, Gary Gaugler wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Alan W Nicholls, PhD Director of Research Service Facility (Electron Microscopy) Research Resources Center - East (M/C 337) Room 100 Science and Engineering South Building The University of Illinois at Chicago 845 West Taylor St Chicago, IL 60607-7058
There is a (relatively) cheaper substitute for Durcupan called Aquembed, available from Ladd. I used to use it routinely for processing alcohol-sensitive material into Epon and it worked very well, though it extends the processing times considerably. I never tried it after DMSO, but I was always careful to wash all traces of fixative and buffers out before going to the first dilution of Aquembed.
Lesley Weston.
on 22/10/2003 12:10 PM, Sergey Ryazantsev at sryazant-at-ucla.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 } ---------------------------------------------------------------------------- --} - } } } } Unfortunatelly, alcohols and acetone dissolve a biological structure that } } I am interested in visualizing. Therefore I am looking for alternative } } solvents. } } } } -Is it possible to fix cells in DMSO?=====} DMSO will not chemically fix } } -Does DMSO interact with OsO4?=====} perhaps no, check Merk Index for } } compatibility. } } -Does it interact with glutaraldehyde?======} see above } } -Is there a resin that mixes with DMSO?====} perhaps no } } Dear Martin } I don't understand your point. How do you know that alcohol and acetone } dissolve your structure? Did you chemically fix your sample before? If } so, it quite unlikely that your "structure" will be "dissolved". I am } surprise: DMSO is such good solvent, it should "dissolve" even better than } ethanol or acetone. Nevertheless, you may try some water-compatible } embedding media to avoid organics. Some acrylates may do the } job. Durcupan also comes to my mind (old stuff). Another choice is to use } "freeze-substitution" - it's shown that at low temperature acetone has much } smaller "dissolving" ability... Good luck, Sergey } }
From MicroscopyL-request-at-ns.microscopy.com Sat Oct 25 09:32:53 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (tpepper-at-iastate.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, October 23, 2003 at 13:29:48 ---------------------------------------------------------------------------
Question: Greetings: I had a client come by today with a bunch of slides made from paraffin sections and some cryo sections of various rat parts that had lost their hematoxylin counter-staining (Gill's formula). Any clues as to what is causing/caused this? They were stored in dark in slide boxes, some slides have sections with stain and some without. It is showing up on slides from this last year and is inconsistent. Very strange! Thanks, Tracey Pepper
Can you change the angular acceptance as you would by changing the camera length on a TEM/STEM ? On a TEM/STEM you can have ADF, HAADF and "medium" or "low" angle ADF, which can be useful (i.e., for strain imaging). I don't know if you can have the same range of options on a SEM/STEM and what the angle(s) are and if they completing eliminate diffraction contrast. Anyone know?
----- Original Message ----- } From: "Alan Nicholls" {nicholls-at-uic.edu} To: "Gary Gaugler" {gary-at-gaugler.com} Cc: "MSA listserver" {Microscopy-at-MSA.Microscopy.Com} Sent: Friday, October 24, 2003 7:47 AM
Call for Nomination of Individuals to be considered for Major Awards by the Microscopy Society of America.
Awards:
Distinguished Scientist Awards:
These Awards recognize preeminent senior scientists from both the Biological and Physical disciplines who have a long-standing record of achievement during their career in the field of microscopy or microanalysis.
Burton Medal:
The Burton Medal was initiated to honor the distinguished contributions to the field of microscopy and microanalysis of a scientist who is less than 40 years of age on January 1st of the award year.
Optical Imaging Association-MSA Outstanding Young Investigator Award:
This Award, initiated in 1999, recognizes the distinguished contributions in the field of optical microscopy made by a scientist who is less than 40 years of age on January 1st of the award year.
Outstanding Technologist Awards:
These Awards honor technologists from both the Biological and Physical Sciences who have made significant contributions such as the development of new techniques which have contributed to the advancement of microscopy and microanalysis.
Morton D. Maser Distinguished Service Award:
This Award was initiated to recognize outstanding volunteer service to the Society as exemplified by Mort Maser, who served the Society for many years with great dedication. This award is made to honor an MSA member who has provided significant volunteer service to the Society over a period of years.
Nomination Requirements:
The Distinguished Scientist, Burton Medal, OIA-MSA Outstanding Young Investigator and Outstanding Technologist Awards Nominations should include:
1) a letter from the primary MSA nominator describing the research accomplishments of the candidate with particular emphasis on the unique technical achievements in the Physical or Biological Sciences; and 2) supplemental letters of support from other members of the scientific community.
The Morton D. Maser Distinguished Service Award Nomination should include:
1) a letter from the primary MSA nominator describing the basis for the nomination; and 2) supplemental letters of support from other members of MSA.
The Deadline for receipt of Awards Nomination Packages is December 15, 2003.
Please contact the MSA Business Office for additional information.
Judy Janes, Administrative Manager Bostrom Corporation 230 E. Ohio Street, Suite 400 Chicago, IL 60611-3265 (800) 538-3672; Fax (312) 644-8557, jjanes-at-MSA.microscopy.org
Thanks,
William T. Gunning, Ph.D. Associate Professor of Pathology Medical College of Ohio Department of Pathology BHSB 140 3035 Arlington Avenue Toledo, Ohio 43614-5806 Phone: 419-383-5256 Fax: 419-383-3066 email: wgunning-at-mco.edu
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 27 07:22:04 2003
Robert, Thales-Optem has couplers for Coolpix and most other compact digital cameras. Our customers have been very successful with them. Check out http://www.thales-optem.com/dca.html versions are available to fit C-mount, eyepiece, and many photoport or phototube mountings. We can assist with configuration if needed.
George
George Laing National Graphic Supply 800.223.7130 x3109 scisales-at-ngscorp.com
} } Listers } My Nikon 990 threaded adapter (on the camera end) has seen its better days } after repeated threading and unthreading (I know shame on me for not } dedicating....budget). It now has no more threads to attach to the } MDC/c-mount adapter. Does anyone out there have a solution to this problem? } Is there a threaded adapter available from Nikon or elsewhere? Any } solution is better than no solution. Thank you }
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 27 10:08:54 2003
Most digital cmaeras I hear of in the consumer market use FireWire (IEEE 1394), but in more professional high-speed applications Cameralink seems to be a better solutions as it has more bandwidth to offer. Does anyone have experience with CameraLink cameras for microsocpy ?
It seems to me that a CameraLink camera and interface are the best choice for the highest speed imaging applications. A CameraLink interface can achieve real-time 30fps 12-bit video at 3.2 Megapixel resolution. In its base configuration, CameraLink can transfer at up to 1.6 Gb/sec. and in full configuration, using 2 cables, the rate is extended up to 4.8 Gb/sec.
With the FireWire cameras I know of you pay a serious speed penalty when capturing at full frame size without binning.
The simple answer is no. The detector is a fixed distance from the specimen and there are no lenses in between. In principle you can change the inner angle by using a different hole size, a mask in front of the detector or a segmented solid state detector made up of many rings - not sure any of these have been used in an SEM, all three have been tried in dedicated 100kV STEMs which also have no post specimen lenses.
Alan
At 08:17 AM 10/25/2003 -0700, Edward Principe wrote: } However, } } Can you change the angular acceptance as you would by changing the camera } length on a TEM/STEM ? } On a TEM/STEM you can have ADF, HAADF and "medium" or "low" angle ADF, which } can be useful (i.e., for strain imaging). } I don't know if you can have the same range of options on a SEM/STEM and } what the angle(s) are and if they completing eliminate diffraction contrast. } Anyone know? } } } ----- Original Message ----- } From: "Alan Nicholls" {nicholls-at-uic.edu} } To: "Gary Gaugler" {gary-at-gaugler.com} } Cc: "MSA listserver" {Microscopy-at-MSA.Microscopy.Com} } Sent: Friday, October 24, 2003 7:47 AM } Subject: [Microscopy] Re: Re: Darkfield STEM } } } } } } } } -------------------------------------------------------------------------- } ---- } } 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 } } -------------------------------------------------------------------------- } ----- } } } } Gary } } } } The geometry, what is and how it is collected is exactly the same as STEM } } in TEM. } } } } The BF/DF detectors are after the specimen and collect electrons that have } } been transmitted through the (thin) specimen. The BF detector collects } } electrons close to the optical axis which have not suffered a scattering } } interaction with the specimen (direct beam). The Annular DF detector, } } which is a ring scintillator with a hole in the center, simultaneously } } collects electrons that have been forward scattered from the specimen in } to } } a particular angular range (typical 10mrad to maybe 300mrad). Close to } the } } direct beam these are diffracted and inelastically scattered electrons, } } further out these are elastically scattered electrons. Assuming the dark } } field image collects most of the scattered electrons the image will look } } like the negative of the bright field image. } } } } The technique is briefly mentioned in "Scanning Electron Microscopy and } } X-ray Microanalysis" by Goldstein et al. and also more extensively in } } "Transmission Electron Microscopy" by Williams and Carter } } } } Alan } } } } At 07:10 PM 10/23/2003 -0700, Gary Gaugler 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 } } } } --------------------------------------------------------------------------- } ---- } } } } } } Thanks for the response. } } } } } } the info I seek is specific to STEM on/in } } } an SEM. Major SEM makers offer a STEM option. } } } Some tough the BF and DF feature. I just don't } } } understand how a dark field STEM image is } } } acquired on a SEM which has a STEM stage option. } } } } } } gary g. } } } } } } } } } At 06:56 PM 10/23/2003, you wrote: } } } } Gary, } } } } } } } } The detectors are the same, the difference is in geometry. Bright field } } } } is a circle, and dark field is a ring around the circle. In } } } } a properly aligned STEM all that is required to switch from BF to DF is } } } } to switch the sensors. Both BF and DF signals are present } } } } all time, it matters what sensor you are acquiring from. I am not sure } } } } what you mean by SEM. This sensor arrangement will produce } } } } results identical for both sensors in SEM. Did you mean TEM? Sensors, } } } } BTW, are same (well, almost) as for SEM backscattered } } } } electrons detectors. I can look up something and fax it to you if you } } } } wish. Several pages. } } } } } } } } Vitaly Feingold } } } } Scientific Instruments and Applications } } } } 2773 Heath Lane, Duluth GA 30096 } } } } (770)232-7785 ph. } } } } (770)232-1791 fax } } } } (678)467-0012 mobile } } } } www.sia-cam.com } } } } } } } } This message is made of 100% recycled electrons. } } } } } } } } This address can not receive messages larger than 15 kb without prior } } } } notification. } } } } } } } } ----- Original Message ----- } } } } From: "Gary Gaugler" {gary-at-gaugler.com} } } } } To: "MSA listserver" {Microscopy-at-MSA.Microscopy.Com} } } } } Sent: Thursday, October 23, 2003 6:31 PM } } } } Subject: [Microscopy] Darkfield STEM } } } } } } } } } } } } } } } } } } } } } } } } } } } } ------------------------------------------------------------------------- } ----- } } } } } 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: } } } } } } } } } } Is there a reference source that explains how } } } } } a BF/DF STEM detector works on a SEM? In } } } } } particular, how does it do DF? } } } } } } } } } } tnx, } } } } } gary g. } } } } } } } } } } } } } } } } } } } } } } } } } Alan W Nicholls, PhD } } Director of Research Service Facility (Electron Microscopy) } } Research Resources Center - East (M/C 337) } } Room 100 Science and Engineering South Building } } The University of Illinois at Chicago } } 845 West Taylor St } } Chicago, IL 60607-7058 } } } } Tel: 312 996 1227 } } Fax: 312 996 8091 } } Office: Room 110 } } } } Web site www.rrc.uic.edu } } } } } }
Alan W Nicholls, PhD Director of Research Service Facility (Electron Microscopy) Research Resources Center - East (M/C 337) Room 100 Science and Engineering South Building The University of Illinois at Chicago 845 West Taylor St Chicago, IL 60607-7058
I agree with Peter, that the Cameralink currently is the faster interface, and for applications that need this speed it is probably the better one. However, for cameralink you need to add a card to your computer or a PCMCIA card to your laptop, whereas IEEE 1394 is nowadays directly build into the computer and operating system. As far as speed is concerned, Firewire in it's present generation allows up to 400 Mbps, and we will soon see 800 and 1.6 Gbps.
At present, many cameras (especially in the TEM and fluorescence sectors) are limited not by the transfer rate of the interface, but by intensity issues. If you want to transfer 30 frames per second, you must also be able to live with exposure times of 30 msec or less. This may only be possible through larger pixels on the chip (=lower resolution) or binning (again lower resolution).
Another factor is, that for higher transfer rates you also need to read out the chip faster. The so-called "readout noise", however, is proportional to the readout speed. This again may lead to the transfer speed being of less importance.
If you have applications that require the high bandwidth (several cameras, or situations where noise and intensity is not a problem), the cameralink interface provides higher throughput. In other cases, the bandwidth may not be the most important factor. I don't think, there is a general answer for this question.
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: Peter Van Osta [mailto:pvosta-at-unionbio-eu.com] Sent: Monday, October 27, 2003 9:14 AM To: Confocal Microscopy List; MSA
Hi,
Most digital cmaeras I hear of in the consumer market use FireWire (IEEE 1394), but in more professional high-speed applications Cameralink seems to be a better solutions as it has more bandwidth to offer. Does anyone have experience with CameraLink cameras for microsocpy ?
It seems to me that a CameraLink camera and interface are the best choice for the highest speed imaging applications. A CameraLink interface can achieve real-time 30fps 12-bit video at 3.2 Megapixel resolution. In its base configuration, CameraLink can transfer at up to 1.6 Gb/sec. and in full configuration, using 2 cables, the rate is extended up to 4.8 Gb/sec.
With the FireWire cameras I know of you pay a serious speed penalty when capturing at full frame size without binning.
I've searched the list-server archives as much as I have patience for, and the 'library' here is a collection of just about every biological book on EM but none seem to document a procedure for making lacey grids.
Before I make the recommendation to purchase grids from one of the EM supply houses, I would like to make sure that making these isn't something that can be done easily and routinely.
The intended application is not for biological use. If anyone has a protocol they can share it would be much appreciated.
Thank you,
Geoff Williams Microscopy Facility Supervisor
CMU Biology Department Microscopy Facility web page. http://www.cst.cmich.edu/centers/microscopy/
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 27 17:58:05 2003
Lacey films are extremely holey films containing numerous holes so that specimens can be suspended over the thin, web-like extensions of plastic or carbon-coated plastic. We describe a method in our textbook (page 95, 2nd edition) using glycerine suspended in Formvar.
Here is another method, described by F.S. Sjostrand in 1956 (Proceedings of First Europena Regional Conference on electron Microscopy, Stockholm, p. 120). This is also described in detail on pages 294-297 of his textbook: Sjostrand, F.S. 1967. Electron Microscopy of Cells and Tissues. Academic Press, 462 pages.
1. Dip microscope slide into solution of 2% Formvar (or Butvar) in ethylene dichloride. 2. IMMEDIATELY transfer the slide to a beaker containing a saturated atmosphere of ethylene dichloride (ED) for 20 min. (Make this by lining a beaker with filter paper and soaking the paper with the ED. Keep tightly covered.) 3. After 20 min, remove cover of ED chamber and blow in a stream of air saturated with water vapor. (Do this by blowing air through a flask of water at about 70 degrees C.) 4. Float the formvar net on a water surface. About the upper 2/3 of the slide has the lacey part of the film. 5. Coat with carbon for strength, if desired.
JB
} I've searched the list-server archives as much as I have patience for, } and the 'library' here is a collection of just about every biological } book on EM but none seem to document a procedure for making lacey grids. } } Before I make the recommendation to purchase grids from one of the EM } supply houses, I would like to make sure that making these isn't } something that can be done easily and routinely. } } The intended application is not for biological use. If anyone has a } protocol they can share it would be much appreciated. } } Thank you, } } Geoff Williams } Microscopy Facility Supervisor -- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu ##############################################################
From MicroscopyL-request-at-ns.microscopy.com Mon Oct 27 21:41:09 2003
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (timothy-at-che.utexas.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, October 27, 2003 at 08:09:50 ---------------------------------------------------------------------------
Question: We are in initial stages of purchasing a cryogenic microtome and would like to gain some insight on the quality and performance of commercial microtomes. The brand of microtomes that we are currently looking at are RMC, Microstar, and Leica. Of these brands, which one would you recommend or not recommend and why? The microtome will be primarily used to cut polymeric samples.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Stefan.Gunnarsson-at-ebc.uu.se) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, October 27, 2003 at 09:19:51 ---------------------------------------------------------------------------
Email: Stefan.Gunnarsson-at-ebc.uu.se Name: Stefan Gunnarsson
We are in the process of buying a FEGSEM with EDS and are thinking about an EDS-system with an LN2-free detector. Both EDAX and Noran claim to have such detectors that compare in performance (regarding resolution and lifetime) with the LN2-cooled ones. If anyone has experience of these things, I would be very happy for a bit of input and opinions on this. (There are several reasons why we don't want LN2: safety, workload, availability etc.)
thanks for any replies,
Stefan
...................................................................................................................... Dr Stefan Gunnarsson Evolutionary Biology Centre Microscopy and Imaging Unit Norbyv”gen 18A SE75236 Uppsala, Sweden Tel & Fax: +46 - 18471 2638 ......................................................................................................................
We here in Botswana have a "in between" detector from EDAX on our ESEM. We are very happy with it. You fill it with LN2 when you need it. Wait for ~30 min to stabilize and use it. Keep LN2 for roughly two days. No forced 365 day a year forced filling even on Christmas we can be free (Was it not for the TEM!) I personally feel uncomfortable to let a crystal to room temp since the "fear of God" was put in me if it does happen. Battling to break the bond on the law. Otherwise it is great. Have no experience with LN2 free detrectors. Just worht having a look at the ressolution and light element sensitivity.
hope it helps
-----Original Message----- } From: by way of MicroscopyListserver [mailto:Stefan.Gunnarsson-at-ebc.uu.se] Sent: Tuesday, October 28, 2003 5:48 AM To: microscopy-at-ns.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Stefan.Gunnarsson-at-ebc.uu.se) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, October 27, 2003 at 09:19:51 ---------------------------------------------------------------------------
Email: Stefan.Gunnarsson-at-ebc.uu.se Name: Stefan Gunnarsson
We are in the process of buying a FEGSEM with EDS and are thinking about an EDS-system with an LN2-free detector. Both EDAX and Noran claim to have such detectors that compare in performance (regarding resolution and lifetime) with the LN2-cooled ones. If anyone has experience of these things, I would be very happy for a bit of input and opinions on this. (There are several reasons why we don't want LN2: safety, workload, availability etc.)
thanks for any replies,
Stefan
..................................................................................................................... Dr Stefan Gunnarsson Evolutionary Biology Centre Microscopy and Imaging Unit Norbyv"gen 18A SE75236 Uppsala, Sweden Tel & Fax: +46 - 18471 2638 .....................................................................................................................
Colleagues, (1) has anybody successfully adapted the NIKON COOLPIX 5400 to a microscope via a C-mount adapter? (2)by use of the c-mount adapter as available for the Coolpix 990 / 995 / 4500 ?? (3)are you content with the results ? Please respond short and informally, at best off-line, to peter.heimann-at-uni-bielefeld.de Thanks, Peter
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 02:39:59 2003
Dear all, Not quite microscopy directly, but is anyone using software to collate and organise references to the literature, which can also export to Word format or that sort of thing. Any hints would be greatly appreciated.
Thanks
-- Ian MacLaren Technische Universität Darmstadt Material- und Geowissenschaften Petersenstr. 23 64287 Darmstadt Germany http://www.tu-darmstadt.de/fb/ms/fg/sf/projekte/maclaren-Dateien/maclaren.html
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 06:24:55 2003
a general problem with digital cameras and microscope adapters is the lens diameter of the camera and the lens diameter of the microscope eye piece/c-mount. If you buy a digital camera from a microscope manufacturer with c- mount adapter, you get a modified eye piece with a c-mount thread and a holder for the camera (which places the lens of the camera in front of the eye piece); Leica sold that a few month ago for app. 2000 € with a Canon S40. For good pictures you need a camera with a small front lens, and an eye piece with a large lens; and still you have to use the zoom. Otherwise you get circular pictures; surrounded with a black frame. Before I bought the Canon, I thought of the Nikon 5400 (b/c of the wide field lens), but then decided for the Canon (because it comes with a nice software "remote capture" to operate the camera with a PC). I think the Nikon 5400 is not the best choice for taking pictures with a microscope, the Nikon 4500 is much better. The Canon G3 is as bad as the 5400, but I bought it not just for microscopy, but also for taking pictures with a ring flash e.g.
:-) Torsten
} } } ---------------------------------------------------------------------- } -------- 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 } ---------------------------------------------------------------------- } --------- } } Colleagues, } (1) has anybody successfully adapted the NIKON COOLPIX 5400 to a } microscope via a C-mount adapter? (2)by use of the c-mount adapter as } available for the Coolpix 990 / 995 / 4500 ?? (3)are you content with } the results ? Please respond short and informally, at best off-line, } to peter.heimann-at-uni-bielefeld.de Thanks, Peter } } }
I like Reference Manager, b/c it has an internal pubmed search. What I don't like is, that these programs only support MS Word directly, and not other formats (anymore), like Lotus Wordpro/Wordperfect/OpenOffice.
:-) Torsten
} --------- } } Dear all, } Not quite microscopy directly, but is anyone using software to collate } and organise references to the literature, which can also export to } Word format or that sort of thing. Any hints would be greatly } appreciated. } } Thanks } } --
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 06:43:39 2003
Dear all, Can anyone explain to me the difference between "Endnote" and "Reference Manager"? They both seem to be made by the same company, cost about the same, but I couldn't extract a comparison of what the two packages do differently from their websites.
Thanks again
-- Ian MacLaren Technische Universität Darmstadt Material- und Geowissenschaften Petersenstr. 23 64287 Darmstadt Germany http://www.tu-darmstadt.de/fb/ms/fg/sf/projekte/maclaren-Dateien/maclaren.html
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 07:47:17 2003
Or, given the amount of effort involved, you could just buy some. I am sure EBS, SPI, Fullam and Pella would be glad to supply some at reasonable cost. If you have never made them before I would have thought that buying was the almost surefire way of getting what you want.
On Monday, October 27, 2003, at 07:03 PM, John J. Bozzola 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 } ----------------------------------------------------------------------- } -------- } } Geoff, } } Lacey films are extremely holey films containing numerous holes so } that specimens can be suspended over the thin, web-like extensions of } plastic or carbon-coated plastic. We describe a method in our textbook } (page 95, 2nd edition) using glycerine suspended in Formvar. } } Here is another method, described by F.S. Sjostrand in 1956 } (Proceedings of First Europena Regional Conference on electron } Microscopy, Stockholm, p. 120). This is also described in detail on } pages 294-297 of his textbook: Sjostrand, F.S. 1967. Electron } Microscopy of Cells and Tissues. Academic Press, 462 pages. } } 1. Dip microscope slide into solution of 2% Formvar (or Butvar) in } ethylene dichloride. } 2. IMMEDIATELY transfer the slide to a beaker containing a saturated } atmosphere of ethylene dichloride (ED) for 20 min. (Make this by } lining a beaker with filter paper and soaking the paper with the ED. } Keep tightly covered.) } 3. After 20 min, remove cover of ED chamber and blow in a stream of } air saturated with water vapor. (Do this by blowing air through a } flask of water at about 70 degrees C.) } 4. Float the formvar net on a water surface. About the upper 2/3 of } the slide has the lacey part of the film. } 5. Coat with carbon for strength, if desired. } } JB } } } } } I've searched the list-server archives as much as I have patience for, } } and the 'library' here is a collection of just about every biological } } book on EM but none seem to document a procedure for making lacey } } grids. } } } } Before I make the recommendation to purchase grids from one of the EM } } supply houses, I would like to make sure that making these isn't } } something that can be done easily and routinely. } } } } The intended application is not for biological use. If anyone has a } } protocol they can share it would be much appreciated. } } } } Thank you, } } } } Geoff Williams } } Microscopy Facility Supervisor } -- } ############################################################## } John J. Bozzola, Ph.D., Director } I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) } 750 Communications Drive - MC 4402 } Southern Illinois University } Carbondale, IL 62901 U.S.A. } Phone: 618-453-3730 } Fax: 618-453-2665 } Email: bozzola-at-siu.edu } ############################################################## }
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 08:14:49 2003
} } Dear all, } Not quite microscopy directly, but is anyone using software } to collate and } organise references to the literature, which can also export to Word } format or that sort of thing. Any hints would be greatly appreciated. } } Thanks } } -- } Ian MacLaren } Technische Universität Darmstadt } Material- und Geowissenschaften } Petersenstr. 23 } 64287 Darmstadt } Germany } http://www.tu-darmstadt.de/fb/ms/fg/sf/projekte/maclaren-Datei en/maclaren.html
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 08:31:11 2003
Microscopy Today, June 1997, Bruce Wagner, "Home Made Holey Film Grids".
Phil
} Dear list, } } I've searched the list-server archives as much as I have patience for, } and the 'library' here is a collection of just about every biological } book on EM but none seem to document a procedure for making lacey grids. } } Before I make the recommendation to purchase grids from one of the EM } supply houses, I would like to make sure that making these isn't } something that can be done easily and routinely. } } The intended application is not for biological use. If anyone has a } protocol they can share it would be much appreciated. } } Thank you, } } Geoff Williams } Microscopy Facility Supervisor } } CMU Biology Department Microscopy Facility web page. } http://www.cst.cmich.edu/centers/microscopy/
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 08:44:45 2003
You can find recipes the the "Tips & Tricks" resource at http://www.biotech.ufl.edu/EM/tips/tem.html
in the section titled "holey grid recipie" (sic) The process is not too difficult especially if you don't mind a bit of variability in the hole size, etc.
Thanks to Greg Erdos, et al, for maintaining the "Tips & Tricks" resource!
Cheers, Henk Colijn
At 03:18 PM 10/27/2003 -0500, Geoff Williams wrote:
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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 28 09:17:34 2003
You might want to try the program, Endnotes. I think that it is just what you want.
-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
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Ian MacLaren [mailto:maclaren-at-tu-darmstadt.de] Sent: Tuesday, October 28, 2003 3:45 AM To: Microscopy Listserver
Dear all, Not quite microscopy directly, but is anyone using software to collate and organise references to the literature, which can also export to Word format or that sort of thing. Any hints would be greatly appreciated.
Thanks
-- Ian MacLaren Technische Universität Darmstadt Material- und Geowissenschaften Petersenstr. 23 64287 Darmstadt Germany http://www.tu-darmstadt.de/fb/ms/fg/sf/projekte/maclaren-Dateien/maclaren.html
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 09:19:36 2003
I use Biblioscape. It's cheaper than some of the others, with all the functions. A few years ago, I researched the various options and decided on Biblioscape. It includes a built-in web browser and MS Word and Wordperfect integration. There is actually a freeware version of the software (Biblioexpress). Here's the web site. http://www.biblioscape.com/ Shannan
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Dear all, Not quite microscopy directly, but is anyone using software to collate and organise references to the literature, which can also export to Word format or that sort of thing. Any hints would be greatly appreciated.
Thanks
-- Ian MacLaren Technische Universität Darmstadt Material- und Geowissenschaften Petersenstr. 23 64287 Darmstadt Germany http://www.tu-darmstadt.de/fb/ms/fg/sf/projekte/maclaren-Dateien/maclaren.html
Shannan Little Research Technician/Technicien de recherche Electron Microscopy and Image Analysis / Microscopie électronique et Analyse d'images Agriculture and Agri-Food Canada/Agriculture et Agroalimentaire Canada Telephone/Téléphone: 403-317-3446 Facsimile/Télécopieur: 403-382-3156 P.O. Box 3000 / CP 3000 Lethbridge, Alberta T1J 4B1 littlesm-at-em.agr.ca http://res2.agr.ca/lethbridge/emia/index_e.htm
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 09:33:56 2003
I can't answer your question but I will make a related comment. One of my grad students badgered me into getting Endnote. I have had similar packages in the past and not thought they were worth the effort since I don't write so many papers that it is a huge burden to enter the references in my hand. Most of the citations were in previous papers and I can cut and paste. And one can use Medline or Pubmed to search and organize the latest references on a topic so I didn't think I would use it. But I have found it very useful. I have begun writing summary notes on every paper I read and storing them in the Comments section on Endnotes. This means I can search for all the papers on a topic and avoid having to pull them and re-read them to get the important facts again. I find that I can write a shorthand summary of the key experimental details and findings in a paragraph. I can note whether I thought the data was impressive or worthless. This has proven to save me a lot of time. In addition, students can log into this file, see my views on the paper and enter their own comments. If I point out a flaw or insightful finding, the students and postdocs can have that info as they review the paper. In summary, I have found Endnote a useful tool. I have no financial interest in this software.
At 01:48 PM 10/28/2003 +0100, you wrote:
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Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Thank you for all the replies... In going through the emails this morning I realized now that maybe the question I posed should have been re-phrased: "I've made holey films before, what is different or is there a difference making lacey carbon films..."
Specifically I've taught/made useable holey films for astigmatism and focus exercises in the TEM classes, but those are small uniform and not terribly numerous and completely unlike images found in some of the references to "lacey carbon films," ie: http://www.personal.rdg.ac.uk/~scsharip/contam.pdf -or- http://www.2spi.com/catalog/grids/cusctgrd.html (i have no reason to display the SPI site other than for example purposes)
For those that are interested: To review the recommendations, there is a significant showing of "just buy them." Many comments recommend breathing on the formvar solution before it dries, or to pass the wet formvar coated slide over boiling water. The second seems to be the same way I've made regular holey films just with more suspend glycerol in the solution, or soap and water in the formvar solution. The final technique that seems promising is the Kuga and Brown method as described by Henk Colijn in the 'tips and tricks' section.
Again thank you all for your suggestions and patience.
Geoff Williams Microscopy Facility Supervisor
Central Michigan University Biology Department Microscopy Facility http://www.cst.cmich.edu/centers/microscopy/
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 10:43:00 2003
I installed a new 50W HBO Hg bulb into my Axiophot's lamp housing. It aligned and worked fine for 4 hrs but the next time I went to re-fire it, the power supply just kept clicking and the bulb wouldn't light. I assumed the bulb mounting had loosened during the first usage so I removed it and re-installed it three times with the same result. I gave up on this bulb but your comments on this secondary problem are welcome. The real problem I would like advice on is the second bulb. It fired up just fine but I have had great difficulty aligning it. I get a good typical primary filament image but can't seem to bring the mirror image into focus. If i close down the fluorescence lamp aperature all the way, I get a small dot for the mirror image and a much brighter band for the real filament image (with brighter tips at the top and bottom of the band). The real image is more like I am used to seeing but I usually get a better mirror image. If I open the fluorescence lamp aperature all the way (where I normally leave it during alignment), I get a diffuse round circle of light for the mirror image. I have tried moving the mirror alignment screws for up down and left right and they are working. I went the full distance of both without much improvement. The focus screw for the mirror goes from all clockwise to all counter clockwise with minor changes in the focus. I did the best alignment I could and get a decent illumination of my sample. But I am wondering if any one can explain why I am not getting a good mirror image or if it is dangerous not to align it better - i was taught that if the two filament images overlap it can result in overheating of the bulb. Thanks for any advice on this. Tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
I'm looking for a pdf or word-document of the following qrticle. Can anyone help me out? Thanks!
Histochemistry. 1985;82(3):205-8. Cowen T, Haven AJ, Burnstock G. Pontamine sky blue: a counterstain for background autofluorescence in fluorescence and immunofluorescence histochemistry.
Sven Terclavers
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 12:22:02 2003
Friday Harbor Labs, a marine biological station in the San Juan Islands affiliated with the University of Washington, has an immediate need for a functional transmission electron microscope for biological work.
As the labs' Philips 300 is currently nonfunctional, any TEM work must be done at the UW campus in Seattle - a ferry boat ride and two hour drive away. We have a core group who would like to pursue TEM projects year round and additional researchers who come to the labs in the summertime.
We would like to find a scope that has been maintained under a service agreement.
Please contact:
Paulette Brunner Center for Cell Dynamics Friday Harbor Labs Friday Harbor, Washington 98250 (206)616-0895 pbrunner-at-u.washington.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 12:46:53 2003
Johns Hopkins University still has an opening for an Electron Microscopy Technician in the Department of Pathology. If anyone is interested or knows anyone, pleace contact me.
Thanks
Lois Anderson Johns Hopkins University Dept. of Pathology Laboratory Manager Electron Microscopy/Immunofluorescence 600 N. Wolfe Street/Pathology 709 A Baltimore, MD 21287 (410) 955-2861/fax (410) 614-7110 landers-at-jhmi.edu
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 13:02:58 2003
Dear Geoff, Many years ago a grad student on my TEM wanted holey carbon films and decided to make his own. We make our usual carbon films by casting colldion dissolved in amyl acetate on water, adding grids, scooping up the film+grids on filter paper, then drying the paper, carbon-coating the collodion film on the grids and then dissolving the collodion for 48 hours in chloroform. To make the holey films, he mixed the collodion liquid, which is the plastic carried in amyl acetate, with one or two drops of Triton X-100, which is a surfactant. I'm not sure if he diluted the collodion with more amyl acetate. He shook the bottle with collodion/amyl acetate/ Triton X-100 until it was foamy, then cast his films from that on water. He said it worked better than the other methods that he tried. You may be able to make the films lacey by shaking harder or longer. I'm sorry I cannot be more specific, but it was long ago and I didn't make them myself. Good luck. Regards, Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "Geoff Williams" {willi1gl-at-cmich.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Monday, October 27, 2003 12:18 PM
Dear list,
I've searched the list-server archives as much as I have patience for, and the 'library' here is a collection of just about every biological book on EM but none seem to document a procedure for making lacey grids.
Before I make the recommendation to purchase grids from one of the EM supply houses, I would like to make sure that making these isn't something that can be done easily and routinely.
The intended application is not for biological use. If anyone has a protocol they can share it would be much appreciated.
Thank you,
Geoff Williams Microscopy Facility Supervisor
CMU Biology Department Microscopy Facility web page. http://www.cst.cmich.edu/centers/microscopy/
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 13:22:09 2003
Cowen T, Haven AJ, Burnstock G. Pontamine sky blue: a counterstain for background autofluorescence in fluorescence and immunofluorescence histochemistry. Histochemistry. 1985;82(3):205-8.
The stain pontamine sky blue (PSB) has been shown to reduce background autofluorescence in catecholamine fluorescence and immunofluorescence histochemical preparations. Using PSB as a counterstain on whole-mount stretch preparations of human mesenteric blood vessels, a medium dense noradrenergic nerve plexus is clearly revealed, which previously had been only partially visible because of background autofluorescence. Image analysis of nerve densities in whole-mount stretch preparations of guinea-pig arteries containing noradrenergic, substance P-, and vasoactive intestinal polypeptide (VIP)-positive nerve plexuses shows that PSB staining does not alter the specific neuronal fluorescence and that it improves image definition.
If you subscribe to Loansome Doc, you can order a pdf copy for a price.
Gary Gill
-----Original Message----- } From: Sven Terclavers [mailto:Sven.Terclavers-at-med.kuleuven.ac.be] Sent: Tuesday, October 28, 2003 11:55 AM To: Microscopy-at-msa.microscopy.com; histonet-at-lists.utsouthwestern.edu
Hi all,
I'm looking for a pdf or word-document of the following qrticle. Can anyone help me out? Thanks!
Histochemistry. 1985;82(3):205-8. Cowen T, Haven AJ, Burnstock G. Pontamine sky blue: a counterstain for background autofluorescence in fluorescence and immunofluorescence histochemistry.
Sven Terclavers
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 13:28:25 2003
Reference manager, I think they are up to version 10 or 11 by now.
Geoff
Ian MacLaren 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, } Not quite microscopy directly, but is anyone using software to collate } and organise references to the literature, which can also export to } Word format or that sort of thing. Any hints would be greatly } appreciated. } } Thanks }
-- -- ********************************************** 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 Tue Oct 28 14:21:27 2003
I want to get a string going about whom many consider the Greatest Microscopist, and research scientist of the 20th Century. After all He and Carl Zeiss worked together pre 1920. And everyone of you have utilized Dr. Rife's science even to this day.
Check this out.
http://www.rife.de/mscope/mscope2.htm
Let's have a sane discussion about why all you geniuses are not following in his footsteps.
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 15:50:55 2003
For the secondary problem, it might not be a problem with the lamp or lamp-house, but with the starter (where you switch on the light). It also happened to me once, I got the same problems, and it turned out that the electrical supply-box, the 'starter', did not function correct anymore (just like a neon-lamp, if the small white starter here doesn't work anymore, it also keeps on clicking and flashing, but will not (or just after a while) start burning.
Your first problem might be a problem of the screw for the lens in fron of the lamphouse. Even though you can turn it complete, does the lens fully moves? The same for the mirror, the screw might work, but does the mirror actually moves? you can always try adding a little drop of oil (WD-40).
And indeed, be careful, I was taught the same: if the two filaments overlap, your lamp gets heated too much and can explode.
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--
From MicroscopyL-request-at-ns.microscopy.com Tue Oct 28 20:33:25 2003
On Thursday, October 23, 2003, at 11:33 AM, Sue Tyler wrote:
} My supervisor has just requested our travel and training needs for the } upcoming year. I am new to TEM and would like to attend some biological } type seminars. Does anyone have any suggestions or recommendations for } future meetings? } Dear Sue, I have found the M&M meetings to be excellent. I'm sure that every session will have several biological TEM sections--your main problem will be choosing among them. Yours, Bill Tivol 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 29 05:12:09 2003
} Dear all, } Not quite microscopy directly, but is anyone using software to collate } and organise references to the literature, which can also export to } Word format or that sort of thing. Any hints would be greatly } appreciated. } } Thanks }
I like Reference Manager, b/c it has a nice internal pubmed search and database function. The others have it, too, but I did not test it. (But you get trialware for all of them). What I don't like is, that these programs only support MS Word directly, and not other formats (anymore), like Lotus Wordpro/Wordperfect11/OpenOffice. Refman, Procite, and Endnote are all from the same company (isiresearchsoft), I don't know why they sell different ones and don't fuse them to a single product. Probably b/c there are "traditionalists" out there, and two out of three where originally for Apple OS?
Isiresearchsoft compares the three products:
http://thomsonisiresearchsoft.com/compare/
Some other links: http://ist.uwaterloo.ca/ew/biblio/index.html http://www.cse.bris.ac.uk/~ccmjs/rmeval.htm (quite old)
I attended my first M& M meeting this summer and did not find much to offer in the way of biological applications. Of course every years program is different so I would certainly continue to consider it as a viable option. MSA however has been an invaluable asset and I recently learned of another society ( The Ultrastructural Pathology Society) that I am going to check out. Here is the website www.sup.ultrakohl.com Hope that helps.
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On Thursday, October 23, 2003, at 11:33 AM, Sue Tyler wrote:
} My supervisor has just requested our travel and training needs for the } upcoming year. I am new to TEM and would like to attend some biological } type seminars. Does anyone have any suggestions or recommendations for } future meetings? } Dear Sue, I have found the M&M meetings to be excellent. I'm sure that every session will have several biological TEM sections--your main problem will be choosing among them. Yours, Bill Tivol 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 29 10:44:44 2003
M&M is an excellent meeting for all - well most! - forms of microscopy. While TEM probably comprises about half the meeting, I'll guarantee that you will learn many useful things from the presentations and posters, not to mention the first-rate trade show of new instruments. Note that it is primarily an instrumentation/techniques oriented meeting that is useful to physical and biological participants alike.
The Ultrastructural Pathology meeting is a very good and much smaller, specialized event and is sharply focused on specific, and generally clinical, research issues. They sometimes have one or two technique papers but no trade show. It is often held in other countries.
Peter Ingram
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-- Peter Ingram Sr. Physicist Adj. Professor of Pathology, Duke University Medical Center Box 90319 LaSalle Street Extension DURHAM NC USA 27708-0319
If you want a "sane discussion" about this man's work how about providing some real evidence of what he has accomplished, that is, where his work has been published. Claims made on a website are just that, claims made on a website. When I went on to the homepage and read about how the FDA and the AMA are conspiring to keep his work out of the medical manistream I got a bit skeptical, but that's just me.
Geoff
Bruce Grosso 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 Wed Oct 29 12:33:49 2003
http://www.sumeria.net/tech/rife2.html The Smithsonian Report
http://www.navi.net/~rsc/seidel.htm#rife Journal of the Franklin Institute Volume 237(2):103-130 (1944) Click on the Rife line at the beginning
} 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 } -------------------------------------------------------------------------- ----- } } } Down here in the Southern Hemisphere it's October 30. } } It's not April 1 up there yet, is it? } } cheers } } rtch } } } } } } } From: "Bruce Grosso" {bgrosso-at-AssetRecovery.Net} } To: {Microscopy-at-msa.microscopy.com} } Subject: [Microscopy] Royal Raymond Rife } Date sent: Tue, 28 Oct 2003 14:26:28 -0800 } } } } } } } ---------------------------------------------------------------------- } } -------- 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 want to get a string going about whom many consider the Greatest } } Microscopist, and research scientist of the 20th Century. After all } } He and Carl Zeiss worked together pre 1920. And everyone of you have } } utilized Dr. Rife's science even to this day. } } } } Check this out. } } } } http://www.rife.de/mscope/mscope2.htm } } } } Let's have a sane discussion about why all you geniuses are not } } following in his footsteps. } } } } } } } } -- } 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 Wed Oct 29 14:16:58 2003
After reviewing Dr. Rife's accomplishments one observation is possible. It was probable the inability to fritter time on the internet which allowed him to accomplish these improbable accomplishments.
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238 ----- Forwarded by Frank Karl/AKR/Degussa-Huels/US on 10/29/2003 03:16 PM -----
Geoff McAuliffe To: Bruce Grosso {bgrosso-at-AssetRecovery.Net} {mcauliff-at-umd cc: Microscopy-at-msa.microscopy.com nj.edu} Subject: [Microscopy] Re: Royal Raymond Rife
The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hi Bruce:
If you want a "sane discussion" about this man's work how about providing some real evidence of what he has accomplished, that is, where his work has been published. Claims made on a website are just that, claims made on a website. When I went on to the homepage and read about how the FDA and the AMA are conspiring to keep his work out of the medical manistream I got a bit skeptical, but that's just me.
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } I want to get a string going about whom many consider the Greatest } Microscopist, and research scientist of the 20th Century. After all He and } Carl Zeiss worked together pre 1920. And everyone of you have utilized Dr. } Rife's science even to this day. } } Check this out. } } http://www.rife.de/mscope/mscope2.htm } } Let's have a sane discussion about why all you geniuses are not following in } his footsteps. } } } } }
-- -- ********************************************** 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 Wed Oct 29 14:47:01 2003
-----Original Message----- } From: Ritchie Sims [mailto:r.sims-at-auckland.ac.nz] Sent: Wednesday, October 29, 2003 1:42 PM To: Microscopy-at-msa.microscopy.com
Sorry to disappoint you all, but I really don't think M&M is the best place to go if one's interest is TEM applied to biological questions. I agree there were a few good talks at this year's meeting in that particular field, but too few to justify the expenditure and the time (5-6 days). There are some very good TEM courses organized around the country that would be more suitable for someone who is simply looking for some basic training in Electron Microscopy of biological systems. including immuno- cytochemistry. One such course is the one organized by Elaine Humphrey and Kent McDonald at the University of British Columbia in June 2004. Courses like this are frequently advertised on the listserver. You could also check the websites of the affiliated local societies listed on the MSA website. These societies organize several meetings per year, some in the form of mini-symposiums. Best,
Marc
On Wednesday, October 29, 2003, at 11:50 AM, Peter Ingram 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 } ----------------------------------------------------------------------- } -------- } } Sue: } } M&M is an excellent meeting for all - well most! - forms of } microscopy. While TEM probably comprises about half the meeting, I'll } guarantee that you will learn many useful things from the } presentations and posters, not to mention the first-rate trade show of } new instruments. Note that it is primarily an } instrumentation/techniques oriented meeting that is useful to physical } and biological participants alike. } } The Ultrastructural Pathology meeting is a very good and much } smaller, specialized event and is sharply focused on specific, and } generally clinical, research issues. They sometimes have one or two } technique papers but no trade show. It is often held in other } countries. } } Peter Ingram } } } } ---------------------------------------------------------------------- } } -------- } } 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 } } ---------------------------------------------------------------------- } } --------- } } } } } } On Thursday, October 23, 2003, at 11:33 AM, Sue Tyler wrote: } } } } } My supervisor has just requested our travel and training needs for } } } the } } } upcoming year. I am new to TEM and would like to attend some } } } biological } } } type seminars. Does anyone have any suggestions or recommendations } } } for } } } future meetings? } } } } } Dear Sue, } } I have found the M&M meetings to be excellent. I'm sure that every } } session will have several biological TEM sections--your main problem } } will be choosing among them. } } Yours, } } Bill Tivol } } 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 } } } -- } Peter Ingram } Sr. Physicist } Adj. Professor of Pathology, } Duke University Medical Center } Box 90319 } LaSalle Street Extension } DURHAM NC USA 27708-0319 } } Tel: (919) 660-2695 } Fax: (919) 660-2671 } e-mail: p.ingram-at-cellbio.duke.edu } http://152.3.54.107/AEM_LAB.html } }
-- 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 Wed Oct 29 16:32:43 2003
I guess I was warned by a friend, fairly enough, that most of you would be dismissive but also that there would be an open mind or two as to the possibility that he (Rife) was right and the dismissive are wrong. I always find that to be true quite often. You overlook the man's accomplishments, his positions he held, his papers, his patents, his relationship with Carl Zeiss, and the Army of doctors, scientists, researchers and allies he had and just sweep all those people away and focus on the smallest (if there really is any) negative and exploit that as reason enough to totally dismiss all the positive documented accomplishments this man has to his credit. and pooh pooh it all with snide commentary as if you have anything to lose by keeping an open mind for just a little while and just for giggles digging a little deeper to see if there is a probability rather than an improbability.
I am not trying to sell Rife to anyone, I was just hoping for a "sane" discussion, with real scientists, between one another of yourselves but I see with some of you that is not possible.
It is too easy to crack wisely and dismiss, than to briefly study and discuss the what if and say instead; impossible.
The one great thing about doing the impossible is there is no competition.
I am just hanging around this site, waiting to sell a microscope and thought I would try to get to know few of you. Not start WWIII.
I do know that the ones of you who are curious and are looking at the "What if it is true?" are going to be the ones who say nothing so they can ponder that rather than imperiously dismiss it as balderdash.
You know the mind is like a parachute! It only works when it is open.
Bruce Grosso www.AssetRecovery.Net, Inc. 67 County Rd. 274 Iuka, MS 38852 662-423-1757 Ph. 662-423-1299 Fx bgrosso-at-AssetRecovery.Net Equipment Recovery Specialists ----- Original Message ----- } From: {Frank.Karl-at-degussa.com} To: {microscopy-at-msa.microscopy.com} Sent: Wednesday, October 29, 2003 12:05 PM
The copy of the journal The Scientist that arrived in my mailbox this afternoon contains a comparison of the major players in the reference manager software arena (http://www.the-scientist.com/yr2003/oct/lcprofile1_031020.html); the review seems a bit cursory, but might be of use to those newly-entering the morass of citation management.
|| Aaron Barnes The Evergreen State College Olympia, WA USA
-----Original Message----- } From: Ian MacLaren [mailto:maclaren-at-tu-darmstadt.de] Sent: Tuesday, October 28, 2003 12:45 AM To: Microscopy Listserver
If that is the way you see my approach, then you have my sincerest and my most humble apologies and many times over and again in spades.
I just wanted to get to know a few of you guys and gals better as I wait to sell my scope. (for what it is worth it is a Zeiss LSM-510 META confocal).
I have to wait for the wheels of academic bureaucracy to turn and I was trying to make some new acquaintances in case I get another scope or useful piece of research equipment to sell one day.
I wish I had the vernacular to engage in a discussion about the differences between light frequencies and cellular wall mortal oscillatory rates as compared to the features of the light microscope he built and how in using it he had the intention of finding:
#1) Cancer was a viral pathogen #2) Each specimen emitted it's own distinct light signature, and this was not limited to the first specimens but after years of study he discovered it applied to virtually every pathogen and living cell. #3) That by discovering it light frequency he could emit the matching sonic (for lack of a better more educated word) frequency thus doing a Crystal glass style shattering of the cell wall and destroying the Pathogen without affecting the surrounding tissue in any way as they had different mortal oscillatory rates.
AND what if it is true? Doesn't anyone want to try it out to see if it is or not? I mean we already do it on gall stones and crystal glass. What is the harm on living tissue samples? What could it hurt to just take a look see?
Bruce Grosso www.AssetRecovery.Net, Inc. 67 County Rd. 274 Iuka, MS 38852 662-423-1757 Ph. 662-423-1299 Fx bgrosso-at-AssetRecovery.Net Equipment Recovery Specialists ----- Original Message ----- } From: "Webster, Paul" {PWebster-at-mail.hei.org} To: {microscopy-at-msa.microscopy.com} ; "'Bruce Grosso'" {bgrosso-at-AssetRecovery.Net} Sent: Wednesday, October 29, 2003 5:25 PM
Hi all,
Our Hitachi H-600 TEM is failing to provide an HV reading at any kV (ie 25, 50, 75, 100). As the microscope is 20 years old I believe the HT cable is the problem. Do you agree? What's the best plan of attack from here? Apart from the HT cable and a dirty gun chamber, are there any other potential causes of this failure? I have never attempted to isolate the HT cable. The vacuum system is fine. I've turned the microscope completely on and off a few times and have vented all compartments to air and back to high vacuum but this has had no effect.
Two days ago I reassembled the rear cosmetic guard that encases the upper vacuum system and the HT cable. Could a slight knock to the cable be enough to finish it off? The microscope was working yesterday, though. I've tried gently wiggling the cable but that gave no response.
The microscope has not been hissing and the HV has appeared quite stable. The only sign that something was not quite right was that some mornings the HV would fail to read at 25kV (but would always read at 50, 75 and 100kV).
In the light of problems encountered by Sarah Ellis (refer to archives) a few months ago I am tempted to clean the gun chamber first (however I doubt if this will fix anything).
Any advice would be gratefully accepted before I contact our institution's electrical engineers.
John Brealey EM Unit The Queen Elizabeth Hospital Institute of Medical & Veterinary Science Adelaide Australia (08) 8222 6612 john.brealey-at-imvs.sa.gov.au
From MicroscopyL-request-at-ns.microscopy.com Wed Oct 29 22:31:48 2003
Geoff Williams asked how to make lacey carbon coated grids. There may be many methods; a number of methods may be found in textbooks. Here's how we do it. The problem with instructions like these is that the devil is in the details, and times, temperatures and concentrations all vary with the season.
Take a glass slide. Clean it thoroughly. Dip it into a solution of Formvar in ethylene dichloride. Remove the slide from the Formvar solution and allow it to dry briefly; our current time is about one second, but there's a "feel" for this step. Insert the slide into the "steam chamber" for several seconds, remove the slide and allow it to dry thoroughly. "Score" the film at the edges of the slide using a fresh razor blade and float the film off the slide onto freshly filtered, deionized water.
The "steam chamber" which we use consists of a beaker of water, sitting on a hot plate and covered with a Maxwell House coffee can which has a hole cut in the side about 4 cm square. We do not know if other brands of coffee cans will work as well. The temperature of the water in the beaker is maintained so that there is visible condensation on the side of the beaker, but the water is not actively boiling.
When you have floated the film onto the water, look at it. If it looks clear, it is probably good. If it looks cloudy, or if wrinkles can be seen, you might as well discard it now. Lay a pattern of the grids of your choice onto the film, and pick the film and the grids up using a piece of filter paper. Allow the film, the grids and the filter paper to dry thoroughly. What you now have is grids which have a damaged Formvar film. The grids are made "lacey" by exposing them to the ethylene dichloride vapors above ethylene dichloride-soaked filter paper in a covered Petri dish for several seconds. The exact time of exposure is determined using a stop watch and inspecting sample grids in the TEM; there is no substitute for this inspection step and the feedback of checking sample grids as you process each batch.
The lacey grids can now be carbon coated. After carbon coating, the Formvar film is removed by placing the grids on chloroform-soaked filter paper overnight. Example grids are examined in the TEM to assure that the film is beam stable and free from artifacts. The finished grids are inspected under a low power light microscope and packaged for shipment.
At each step, it is necessary for the specific laboratory to determine the precise concentration, time, temperature, etc. which is required for each step; there is no universal recipe, and the recipe for our laboratory varies from day to day. There is no substitute for inspection of sample grids at each step of the process using a TEM; you simply can't see what you need to see by light microscopy. The cornerstone of our quality assurance program for lacey carbon and other custom coated grids is inspecting example grids from each batch in our own TEM. If the grids do not meet our quality standards, the customer never sees them. This may explain why we have virtually no returns of coated grids from customers.
And it helps to have been doing the process for several decades. This is a skilled art, and the only substitute for going through the learning curve yourself is to ask someone who has "been there" before you to make the grids you need.
Disclaimer: SPI Supplies sells all of the materials necessary to do this except the TEM itself. We also sell lacey carbon coated grids. We have an obvious interest in promoting this technology.
Andy
Andrew W. Blackwood, Ph.D. Vice President, Technical SPI Supplies P.O. Box 656 West Chester, PA 19381-0656 Ph: 1 610 436 5400 X108 FAX: 1 610 436 5755 e-mail: ablackwood-at-2spi.com WWW: http://www.2spi.com
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 30 09:02:26 2003
Thank you for sharing your instructions. We make them a bit differently here but what works for one may not always work for another. I think that sharing your instructions was particularly generous in that you obviously make these grids for sale. However, I suspect that those that typically buy them will continue to do so and those that struggle to make them themselves will just have a bit easier time of it.
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
On 10/30/03 7:43 AM, "Blackwood, Andrew" {ablackwood-at-2spi.com} wrote:
} Geoff Williams asked how to make lacey carbon coated grids. There may be } many methods; a number of methods may be found in textbooks. Here's how we } do it. The problem with instructions like these is that the devil is in the } details, and times, temperatures and concentrations all vary with the } season. } } Take a glass slide. Clean it thoroughly. Dip it into a solution of Formvar } in ethylene dichloride. Remove the slide from the Formvar solution and allow } it to dry briefly; our current time is about one second, but there's a } "feel" for this step. Insert the slide into the "steam chamber" for several } seconds, remove the slide and allow it to dry thoroughly. "Score" the film } at the edges of the slide using a fresh razor blade and float the film off } the slide onto freshly filtered, deionized water. } } The "steam chamber" which we use consists of a beaker of water, sitting on a } hot plate and covered with a Maxwell House coffee can which has a hole cut } in the side about 4 cm square. We do not know if other brands of coffee cans } will work as well. The temperature of the water in the beaker is maintained } so that there is visible condensation on the side of the beaker, but the } water is not actively boiling. } } When you have floated the film onto the water, look at it. If it looks } clear, it is probably good. If it looks cloudy, or if wrinkles can be seen, } you might as well discard it now. Lay a pattern of the grids of your choice } onto the film, and pick the film and the grids up using a piece of filter } paper. Allow the film, the grids and the filter paper to dry thoroughly. } What you now have is grids which have a damaged Formvar film. The grids are } made "lacey" by exposing them to the ethylene dichloride vapors above } ethylene dichloride-soaked filter paper in a covered Petri dish for several } seconds. The exact time of exposure is determined using a stop watch and } inspecting sample grids in the TEM; there is no substitute for this } inspection step and the feedback of checking sample grids as you process } each batch. } } The lacey grids can now be carbon coated. After carbon coating, the Formvar } film is removed by placing the grids on chloroform-soaked filter paper } overnight. Example grids are examined in the TEM to assure that the film is } beam stable and free from artifacts. The finished grids are inspected under } a low power light microscope and packaged for shipment. } } At each step, it is necessary for the specific laboratory to determine the } precise concentration, time, temperature, etc. which is required for each } step; there is no universal recipe, and the recipe for our laboratory varies } from day to day. There is no substitute for inspection of sample grids at } each step of the process using a TEM; you simply can't see what you need to } see by light microscopy. The cornerstone of our quality assurance program } for lacey carbon and other custom coated grids is inspecting example grids } from each batch in our own TEM. If the grids do not meet our quality } standards, the customer never sees them. This may explain why we have } virtually no returns of coated grids from customers. } } And it helps to have been doing the process for several decades. This is a } skilled art, and the only substitute for going through the learning curve } yourself is to ask someone who has "been there" before you to make the grids } you need. } } Disclaimer: SPI Supplies sells all of the materials necessary to do this } except the TEM itself. We also sell lacey carbon coated grids. We have an } obvious interest in promoting this technology. } } Andy } } Andrew W. Blackwood, Ph.D. } Vice President, Technical } SPI Supplies } P.O. Box 656 } West Chester, PA 19381-0656 } Ph: 1 610 436 5400 X108 } FAX: 1 610 436 5755 } e-mail: ablackwood-at-2spi.com } WWW: http://www.2spi.com }
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 30 11:39:51 2003
One of our lab members was looking for bimetallic grids: Cu on one side and Ni on the other. She couldn't remember the supplier, but remembered using them -- within the last couple of years. I've gone on the web and checked all the usual suspects to no avail. Has any one heard of such grids, and knows who supplies them? Thanks.
Phil -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 30 11:45:01 2003
It's very interesting to read all about how the SPI grids we purchase are made...takes me back to my student days! You're absolutely correct that there is an art associated with each step, which varies from lab to lab. That's why it is far more cost effective for my lab to purchase the SPI lacey film products rather than attempt to make them ourselves any more. Of course, we don't have an unlimited supply of graduate students either... ;-)
Larry
Disclaimer: I have no connection whatsoever with SPI Co., and am pleased to pay full price for any product that I purchase from them.
From MicroscopyL-request-at-ns.microscopy.com Thu Oct 30 15:13:21 2003
Dear Phil, I have seen Cu-Pd grids, which Marivac sells, and they look copper-coloured on one side and grey on the other, but I have never seen Cu-Ni.
----- Original Message ----- } From: "Philip Oshel" {peoshel-at-wisc.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, October 30, 2003 9:45 AM
Hi:
Forgive me if this is too simple. Some guys from another lab just came by and asked if I had any quartz coverslips. They are doing Raman microscopy with infrared/red light and they say the ordinary glass coverslips are killing their signal.
I volunteered to check around for them, this is my first stop since, of course, I couldn't think of anyplace else where I could tap directly into the hearts and minds of the world's greatest and most renowned microscopists.
Is there an easy solution to their problem? Inquiring minds want to know. I will check around elsewhere, but I always think of this list whenever questions like this come up.
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 Thu Oct 30 15:53:37 2003
There are a number of reasons why the HT will not switch on.
1. The Vacuum safety trip is not working preventing the HT system operating. 2. The electronics for HT generation are faulty 3. Not so sure that a duff HT cable would give such a problem.
Without a circuit diagram it is difficult for me to tell you how to overcome (1) other than to say I have shorted out the trip in the past in order to get a customer going.
Number (2) could be a lack of a standard supply but always ask "what did I do last?" i.e. prior to the fault. Did you knock one of the connections to the tank or on the microscope end of the system; check their tightness?
Regarding (3) how to check out the cable problems
1. Clean the top of the tank so that there will be no danger of dirt falling into the oil when you open it up. 2. Remove the high voltage cable from the tank and wrap it in clean aluminium foil. Cover the tank opening with foil. 3. Now switch on the HT 4. Still nothing then you have a supply or connection problem, the circuitry is at fault. 5. All is well (I doubt in your case) you have a cable problem 6. Make contact with a local x-ray or high voltage company (nothing to do with EM but will be able to handle a 125kV cable) who should be able to use your cable ends and make a new cable at a fraction of the Hitachi costs, or any one else for that matter!
Hope this helps. I am away for the next two weeks but come back to me if you need more help?
Good luck
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Mob 07711 606967 www.emcourses.com
----- Original Message ----- } From: "John Brealey" {john.brealey-at-imvs.sa.gov.au} To: "Listserver" {Microscopy-at-MSA.Microscopy.Com} Sent: Thursday, October 30, 2003 3:54 AM
Hi John
There are a number of reasons why the HT will not switch on.
1. The Vacuum safety trip is not working preventing the HT system operating. 2. The electronics for HT generation are faulty 3. Not so sure that a duff HT cable would give such a problem.
Without a circuit diagram it is difficult for me to tell you how to overcome (1) other than to say I have shorted out the trip in the past in order to get a customer going.
Number (2) could be a lack of a standard supply but always ask "what did I do last?" i.e. prior to the fault. Did you knock one of the connections to the tank or on the microscope end of the system; check their tightness?
Regarding (3) how to check out the cable problems
1. Clean the top of the tank so that there will be no danger of dirt falling into the oil when you open it up. 2. Remove the high voltage cable from the tank and wrap it in clean aluminium foil. Cover the tank opening with foil. 3. Now switch on the HT 4. Still nothing then you have a supply or connection problem, the circuitry is at fault. 5. All is well (I doubt in your case) you have a cable problem 6. Make contact with a local x-ray or high voltage company (nothing to do with EM but will be able to handle a 125kV cable) who should be able to use your cable ends and make a new cable at a fraction of the Hitachi costs, or any one else for that matter!
Hope this helps. I am away for the next two weeks but come back to me if you need more help?
Good luck
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 Mob 07711 606967 www.emcourses.com
----- Original Message ----- } From: "John Brealey" {john.brealey-at-imvs.sa.gov.au} To: "Listserver" {Microscopy-at-MSA.Microscopy.Com} Sent: Thursday, October 30, 2003 3:54 AM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (scanning-at-fams.org) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, October 30, 2003 at 14:20:26 ---------------------------------------------------------------------------
Organization: SCANNING, The Journal of Scanning Microscopies
Title-Subject: [Microscopy] SCANNING 2004 - Washington, D.C. April 27-29
Question: Dear Microscopists and Friends:
The 15th annual scientific meeting on scanning microscopies, sponsored by the Foundation for Advances in Medicine and Science (FAMS, Inc.), promises to be an exciting and informative scientific meeting on the scanning microscopies. We hope you will join us there to commemorate the first century of scanning electron microscopy and the centennial anniversary of Sir Charles Oatley, pioneer of scanning electron microscopy.
Details of the Meeting:
What: SCANNING 2004 When: Tuesday, April 27, Wednesday, April 28, Thursday, April 29 Where: Hotel Washington, Washington, D.C. Contact: (201) 818-1010 (Tel) (201) 818-0086 (Fax)
Please visit us at www.scanning.org to download the Call for Papers and for the latest information pertaining the the sessions, short courses, hotel information, and sponsorship opportunities.
If that type of bulb is elongated with a terminal at each end, reverse the polarity. i.e. Flip it around and reinsert it. The starter electrode needs to be at the + anode terminal to work. Bob.
Bob Carter 2000 Bayshore Road Lopez Island, WA 98261
----- Original Message ----- } From: "Tom Phillips" {phillipst-at-missouri.edu} } To: {Microscopy-at-msa.microscopy.com} } Sent: Tuesday, October 28, 2003 8:50 AM } Subject: [Microscopy] Hg bulb alignment } } } } } } } } -------------------------------------------------------------------------- } ---- } } 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 installed a new 50W HBO Hg bulb into my Axiophot's lamp housing. It } } aligned and worked fine for 4 hrs but the next time I went to re-fire it, } } the power supply just kept clicking and the bulb wouldn't light. I } assumed } } the bulb mounting had loosened during the first usage so I removed it and } } re-installed it three times with the same result. I gave up on this bulb } } but your comments on this secondary problem are welcome. The real problem } } I would like advice on is the second bulb. It fired up just fine but I } } have had great difficulty aligning it. I get a good typical primary } } filament image but can't seem to bring the mirror image into focus. If i } } close down the fluorescence lamp aperature all the way, I get a small dot } } for the mirror image and a much brighter band for the real filament image } } (with brighter tips at the top and bottom of the band). The real image is } } more like I am used to seeing but I usually get a better mirror image. If } I } } open the fluorescence lamp aperature all the way (where I normally leave } it } } during alignment), I get a diffuse round circle of light for the mirror } } image. I have tried moving the mirror alignment screws for up down and } } left right and they are working. I went the full distance of both without } } much improvement. The focus screw for the mirror goes from all clockwise } } to all counter clockwise with minor changes in the focus. I did the best } } alignment I could and get a decent illumination of my sample. But I am } } wondering if any one can explain why I am not getting a good mirror image } } or if it is dangerous not to align it better - i was taught that if the } two } } filament images overlap it can result in overheating of the bulb. Thanks } } for any advice on this. Tom } } } } 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 } } } } } } }
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 31 02:33:13 2003
} Hello Peter, } } you are perfectly right with your assessment of FireWire cameras. What you } have neglected is the benefit of IEEE1394 being a widespread standard } interface on Apple Macintosh as well as Microsoft Windows computer } platforms. On current computers you do not need expensive framegrabber } boards (e.g. for use with CameraLink) because they are already equipped with } FireWire. This greatly simplifies the usage of one camera in an environment } with more than one microscope wokstation. Furthermore, direct digitization } in the camera's head also minimizes noise that may be introduced to your } signal while beeing transmitted to the framegrabber. So the trade-off is } flexibility vs. speed. } } What do you think? } } Best regards, } } Achim } } } =============================================== } Achim Zimmermann, MSc (Phys) } Product Manager Microscope Cameras } } JENOPTIK Laser, Optik, Systeme GmbH } Business Unit Sensor Systems - Digital Cameras } Göschwitzer Str. 25 } D-07745 Jena } Germany } } Tel.: +49 36 41 65 - 21 39 (office) } Tel.: +49 17 33 99 35 45 (mobile) } Fax: +49 36 41 65 - 21 44 } } eMail: achim.zimmermann-at-jenoptik.com } Web: www.progres-camera.com - www.jenoptik.com } =============================================== } } } } } -----Original Message----- } } From: Peter Van Osta [mailto:pvosta-at-unionbio-eu.com] } } Sent: Monday, October 27, 2003 5:14 PM } } To: Confocal Microscopy List; MSA } } Subject: [Microscopy] Firewire or CameraLink ? } } } } } } } } } } -------------------------------------------------------------- } } ---------------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society } } of America
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 31 02:41:53 2003
We are currently testing our new system in which we use a 75 W Xenon Arc (Carl Zeiss). We have developed a remote control system through which we can control the the power (unipolar switches) from within our software.
Although we got the message that the Xenon Arc is properly shielded it frequently brings other electronic components in the system in disarray and this leads to malfunction of the entire system.
We have tried netfilters and grounding but up to now we have not been able to protect our other equipment from periodic failure due to the spikes in the electricity net and/or the RF generated by the the Xenon Arc.
We take care to schedule the powerup of the Xenon Arc before all other equipment and wait for 15 sec. before proceeding with the powerup sequence. Powerdown is in reverse order and the Xenon Arc is powered down as the last device.
Quartz slides and coverslips are available in various sizes / thicknesses and shapes from Agar Scientific http://www.agarscientific.com ========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 (0) 131 650 5554 FAX. #44 (0) 131 650 5392 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 31 07:42:59 2003
Try SPI (www.spi2.com). I know that Chuck Garber understands this problem well.
Thanks, Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
~-at-~-at-~~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at- Optimizing Light Microscopy for Biological and Clinical Labs is available in individual copies or classroom size orders. Visit www.MicroscopyEducation.com for details. ~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-~-at-
At 01:23 PM 10/30/03 -0800, Jon Krupp wrote:
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From MicroscopyL-request-at-ns.microscopy.com Fri Oct 31 09:13:32 2003
Jonathan Krupp wrote: ====================================================== Hi:
Forgive me if this is too simple. Some guys from another lab just came by and asked if I had any quartz coverslips. They are doing Raman microscopy with infrared/red light and they say the ordinary glass coverslips are killing their signal.
I volunteered to check around for them, this is my first stop since, of course, I couldn't think of anyplace else where I could tap directly into the hearts and minds of the world's greatest and most renowned microscopists
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 31 11:10:27 2003
Please forgive this attempt to contact Gordon Couger who posted a message regarding scanning light photomacrography on the 21st of October. I have tried several times to email him directly and my messages bounce - they say couger.com is not a local host, not a gateway (whatever that means). Anyway, if I can get a valid email address for Gordon, I will take this off the list. Again, my apologies for dealing with this on the list.
Regards, Bill Sharp
William P. Sharp Arizona State University School of Life Sciences, box 4501 Tempe, AZ 85287-4501 Phone - (480)-965-3210 Fax - (480)-965-6899
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 31 12:30:17 2003
Listers, I would like to get some insight on anybody who is currently using the fluorescence illumination system from the EXFO company's X-Cite 120 (Exfo.com) particularly on the following aspects. Ease of use, User friendliness, stability of the lamp intensity, life of the lamp and the liquid light guide (influence of its liquid filled model, diameter and the claim on 'less heat more light'), and the cost to efficiency ratio or any relevant information on this product. Although some of this information already I am aware of from the manufacturer, but would like to hear from somebody who is currently using it and having hands on experience. Thanks in advance Shiv
Mayandi Sivaguru, Ph.D., Associate Director Molecular Cytology Core Facility Molecular Biology Program 2, Tucker Hall University of Missouri Columbia, MO 65211-7400 sivagurum-at-missouri.edu
Voice: 1-573-882-4895 Fax: 1-573-882-0123
www.biotech.missouri.edu/mcc/
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 31 13:44:26 2003
Does anyone know the recommended floor space suitable for an electron microscopy lab? I'm interested in the requirements for both research and also hospital labs space requirements - or at least the recommended square footage requirements.
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 31 14:50:45 2003
I'm assuming you mean for the microscope itself. Sorry for the U.S. units but we have a 2010F that fits well into a lab with 366 sq. ft., and a Tecnai 30 that fits into a lab with 317 sq. ft. That includes room for sample loading areas (two areas in each case), sample storage and storage of misc. parts and extras in a cabinet.
Our sample prep rooms are a similar size to the microscope rooms.
I hope this helps.
cheers, John
******** John S. Vetrano Sr. Research Scientist Materials Structure and Performance Group Pacific Northwest National Laboratory MSIN P8-16 P.O. Box 999 Richland, WA 99352 Phone: (509)372-0724 Fax: (509)376-6308 Email: mailto:john.vetrano-at-pnl.gov
} ---------- } From: Garry Burgess } Sent: Friday, October 31, 2003 11:49 AM } To: MSA } Subject: [Microscopy] Space Requirements for EM lab } } } } ------------------------------------------------------------------------------ } 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 } ------------------------------------------------------------------------------- } } Does anyone know the recommended floor space suitable for an electron } microscopy lab? I'm interested in the requirements for both research and } also hospital labs space requirements - or at least the recommended square } footage requirements. } } }
From MicroscopyL-request-at-ns.microscopy.com Fri Oct 31 18:30:31 2003
Jonathan Krupp wrote: ====================================================== Hi:
Forgive me if this is too simple. Some guys from another lab just came by and asked if I had any quartz coverslips. They are doing Raman microscopy with infrared/red light and they say the ordinary glass coverslips are killing their signal.
I volunteered to check around for them, this is my first stop since, of course, I couldn't think of anyplace else where I could tap directly into the hearts and minds of the world's greatest and most renowned microscopists
From MicroscopyL-request-at-ns.microscopy.com Sat Nov 1 08:23:39 2003
You can not use glass for this kind of application and SPI Supplies has offered a full range of quartz slides and cover slips designed to solve this problem on URL http://www.2spi.com/catalog/ltmic/quartz.shtml
They are generally available from stock for overnight shipment worldwide.
One should note that, like mica, if you don't know the grade you are purchasing, there is no way to tell what you actually do have. So from the above page, you can see that we feature GE 124 quartz in the fabrication of our quartz slides and cover slips. And full transmittance data is disclosed. See URL http://www.2spi.com/catalog/ltmic/quartz-slides-coverslips.html
Whoever you do purchase from, if not from SPI, make sure you know what quartz you are getting and its characteristics.
One further comment: For the near infrared, MgO might be a better alternative. See URL http://www.2spi.com/catalog/submat/magnesium-oxide.shtml and to the graph showing the transmittance spectrum for MgO.
The absorption curves are given for both materials so one can match product selection with objectives in terms of what part of the spectrum they need their greatest transparency.
Disclaimer: SPI Supplies has offered a full range of quartz slides and cover slips to those working in the UV and near infrared range for some number of years. We over standard shapes and sizes but can also produce custom shapes and sizes.
Chuck
============================================
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