} We are comparing dye sub printers [...]. It is my understanding } that both produce good quality, gray-scale prints (versus } dithered, inkjet prints). ...
Inkjet printers may dither, but you cannot see the dithering at all with modern printers. For excellent color, excellent grayscale, as well as archival prints, I would suggest you take a look at the 9-ink HP Photosmart 8750. It's only downside is a thirst for ink cartridges, but considering the prices you are comparing, this printer would be a bargain.
Genuinely, Michael Shaffer :o) SEM/MLA Lab Coordinator (709) 737-6790 (Ofc) (709) 737-6790 (Lab) {www.micro-investigations.com} {www.esd.mun.ca/epma/} Inco Innovation Centre Memorial University of Newfoundland St. John's, Newfoundland
From MicroscopyL-request-at-ns.microscopy.com Fri Jul 1 07:22:43 2005
We have a Canon i9900. The prints are outstanding and in a way, sadly better than anything I could do in the darkroom.
Alan Stone ASTON
At 07:13 AM 7/1/2005, you wrote:
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Alan Stone ASTON Metallurgical Services Co., Inc. 200 Larkin Drive Ste A Wheeling, IL 60090 847/353-8100 www.astonmet.com
From MicroscopyL-request-at-ns.microscopy.com Fri Jul 1 08:08:20 2005
For a few years Pictrography was my printer of choice (average cost per print is about $2 or $3). Half a year ago I bought cheap inkjet HP Deskjet 6540 for draft prints. Surprisingly, it produces grayscale prints of very good quality, so now I practically stopped using Pictrography.
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} -----Original Message----- } From: John J. Bozzola [mailto:bozzola-at-siu.edu] } Sent: Thursday, June 30, 2005 4:14 PM } To: Microscopy-at-msa.microscopy.com } Subject: [Microscopy] dye sub versus Pictrography } } } } } -------------------------------------------------------------- } ---------------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ----------------- } } We are comparing dye sub printers versus the Fuji Pictrography } PG4500. The price differential is a staggering $22,000 versus $6,500, } respectively. It is my understanding that both produce good quality, } gray-scale prints (versus dithered, inkjet prints). Why the price } differential? I would welcome any comments, user experiences, etc. } Does anyone know the average cost per 8.5 x 11in print? } } Thank you. } -- } ############################################################## } John J. Bozzola, Ph.D., Director } I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) } 750 Communications Drive - MC 4402 } Southern Illinois University } Carbondale, IL 62901 U.S.A. } Phone: 618-453-3730 } Email: bozzola-at-siu.edu } ############################################################## } }
From MicroscopyL-request-at-ns.microscopy.com Fri Jul 1 08:37:32 2005
I have to agree with the ink jet crowd. I have a Canon i9100 for personal use which will make prints up to 19x13 inches and it also gives me much more control than I had in the darkroom (and John Bozzola knows what a switch it is to hear that from me---I used to live in darkrooms!). I simply couldn't justify paying thousands of dollars for high-end dye-sub printers or the Fuji system in our lab, when a $500-600 printer will give excellent quality prints with a life span of 100+ years, using the right inks and papers.
I honestly don't even remember when we made the last print for our users in the lab. I bought a cheaper, but good quality ink-jet a couple of years ago for people who wanted prints and the original ink cartridges are still in it. We shoot film on the TEM and scan it and our SEM takes digital images directly. We give our clients the images on disks and they're happy. Occasionally we do quick work prints on a laser printer upon request, but even this is rare.
I still have a darkroom and two enlargers in storage in my garage, but it's becoming more out of nostalgia than any real hope I'll set them up again someday. I just can't let go......
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
-----Original Message----- } From: Alan Stone [mailto:as-at-astonmet.com] Sent: Friday, July 01, 2005 7:22 AM To: microscopy-at-microscopy.com
We have a Canon i9900. The prints are outstanding and in a way, sadly better than anything I could do in the darkroom.
Alan Stone ASTON
At 07:13 AM 7/1/2005, you wrote:
} ----------------------------------------------------------------------- } ------- The Microscopy ListServer -- Sponsor: The Microscopy Society } of America To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver
} } both produce good quality, gray-scale prints (versus dithered, } } inkjet prints). ... } } Inkjet printers may dither, but you cannot see the dithering at all } with modern printers. For excellent color, excellent grayscale, as } well as archival prints, I would suggest you take a look at the 9-ink } HP Photosmart 8750. It's only downside is a thirst for ink cartridges,
} but considering the prices you are comparing, this printer would be a bargain. } } Genuinely, Michael Shaffer :o) } SEM/MLA Lab Coordinator } (709) 737-6790 (Ofc) } (709) 737-6790 (Lab) } {www.micro-investigations.com} } {www.esd.mun.ca/epma/} } Inco Innovation Centre } Memorial University of Newfoundland } St. John's, Newfoundland
Alan Stone ASTON Metallurgical Services Co., Inc. 200 Larkin Drive Ste A Wheeling, IL 60090 847/353-8100 www.astonmet.com
From MicroscopyL-request-at-ns.microscopy.com Fri Jul 1 09:02:28 2005
There is no reason to spend this kind of money to get excellent B&W prints. Many fine art photographers are making beautiful prints with ink jet printers. Check out Lyson.com or inkjetmall.com. Also, continuous inking systems are available that bypass individual ink cartridges. I will post more specifics later or early next week.
Geoff
John J. Bozzola wrote:
} ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } We are comparing dye sub printers versus the Fuji Pictrography PG4500. } The price differential is a staggering $22,000 versus $6,500, } respectively. It is my understanding that both produce good quality, } gray-scale prints (versus dithered, inkjet prints). Why the price } differential? I would welcome any comments, user experiences, etc. } Does anyone know the average cost per 8.5 x 11in print? } } Thank you.
-- -- ********************************************** 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 Jul 1 10:53:09 2005
ELECTRON MICROSCOPE TECHNICIAN - RESEARCH TRIANGLE PARK (RTP), NORTH CAROLINA AREA
We have an immediate need for a full-time Electron Microscope Technician (maintenance and operation). 3+ years TEM and SEM experience in biological, polymer, carbon/carbon composite, and semi-conductor electron microscopy required. Send resume and salary requirements to sjeffers-at-dgisrd.com.
From MicroscopyL-request-at-ns.microscopy.com Fri Jul 1 11:15:37 2005
This post is tangential to the dye sub vs ink jet discussion. When one needs to compare 2 or more images simultaneously, prints can easily be laid out on a table, but this is not so easy with on-screen display of digital images. One thing that we have done in our lab is to install a second monitor at some workstations and increase the Windows desktop size so that it spans the two monitors. This is satisfactory for AFM images where the basic pixel count is 512x512 but may not be so good with other formats. I am curious to learn what other people do.
When you give your customers digital images only, do you feel there is a risk they could miss an important comparison?
regards, Don Chernoff ================================== Advanced Surface Microscopy, Inc. E-Mail: donc-at-asmicro.com 3250 N. Post Rd., Ste. 120 Voice: 317-895-5630 INDIANAPOLIS IN 46226 USA Toll free: 800-374-8557 (in USA & Canada) web: http://www.asmicro.com Fax: 317-895-5652 [business activities: analytical services in AFM, AFM probes, consulting, training, calibration and test specimens, calibration and measurement software, used NanoScope equipment.]
From MicroscopyL-request-at-ns.microscopy.com Fri Jul 1 19:35:59 2005
Don't forget to use RIP software for excellent tonal range. I've seen photos done with and without and there is a discernible difference and improvement. Damian
-----Original Message----- } From: Geoff McAuliffe [mailto:mcauliff-at-umdnj.edu] Sent: Friday, July 01, 2005 9:01 AM To: John J. Bozzola Cc: Microscopy-at-msa.microscopy.com
There is no reason to spend this kind of money to get excellent B&W prints. Many fine art photographers are making beautiful prints with ink jet printers. Check out Lyson.com or inkjetmall.com. Also, continuous inking systems are available that bypass individual ink cartridges. I will post more specifics later or early next week.
Geoff
John J. Bozzola wrote:
} -------------------------------------------------------------------------- ---- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------------------- ----- } } } We are comparing dye sub printers versus the Fuji Pictrography PG4500. } The price differential is a staggering $22,000 versus $6,500, } respectively. It is my understanding that both produce good quality, } gray-scale prints (versus dithered, inkjet prints). Why the price } differential? I would welcome any comments, user experiences, etc. } Does anyone know the average cost per 8.5 x 11in print? } } Thank you.
-- -- ********************************************** 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 Sat Jul 2 02:54:09 2005
Don; I tried to purchase an IBM T-220 9 megapixel display, but was told by IBM that they had killed the product. This was the only display ever marketed that could display a 2K X 2K camera image showing all of the pixels on one screen. We still have two screens on our TEM, but need to either display the images at half resolution, or zoom to display only part of the image.
John Mardinly
-----Original Message----- } From: Don Chernoff at ASM [mailto:donc-at-asmicro.com] Sent: Friday, July 01, 2005 9:05 AM To: Microscopy List
This post is tangential to the dye sub vs ink jet discussion. When one needs to compare 2 or more images simultaneously, prints can easily be laid out on a table, but this is not so easy with on-screen display of digital images. One thing that we have done in our lab is to install a second monitor at some workstations and increase the Windows desktop size so that it spans the two monitors. This is satisfactory for AFM images where the basic pixel count is 512x512 but may not be so good with other formats. I am curious to learn what other people do.
When you give your customers digital images only, do you feel there is a risk they could miss an important comparison?
regards, Don Chernoff ================================== Advanced Surface Microscopy, Inc. E-Mail: donc-at-asmicro.com 3250 N. Post Rd., Ste. 120 Voice: 317-895-5630 INDIANAPOLIS IN 46226 USA Toll free: 800-374-8557 (in USA & Canada) web: http://www.asmicro.com Fax: 317-895-5652 [business activities: analytical services in AFM, AFM probes, consulting, training, calibration and test specimens, calibration and measurement software, used NanoScope equipment.]
From MicroscopyL-request-at-ns.microscopy.com Sat Jul 2 03:04:41 2005
John; We have a Fuji Pictrograph 3500, and I have to say that nothing I have ever seen prints with the resolution and vivid saturation of the pictrograph, both for color and B&W. However, since all of our conference rooms got digital projectors, we don't make prints any more. I used it to make some absolutely beautiful prints of my daughter, but that's about all. We have had multiple failures of a $1,000 circuit board, and the Fuji service department is a NIGHTMARE to deal with. Right now, the printer is inoperative. The most recent batch of paper and donor sticks to the drum, causing a fault. I don't know if spending another $400 for new rolls will cure that problem, or if there is something else wrong with the printer, but we're probably going to just push it out the door.
John Mardinly Intel
This is the opinion of the author and not of Intel Corporation.
-----Original Message----- } From: John J. Bozzola [mailto:bozzola-at-siu.edu] Sent: Thursday, June 30, 2005 2:14 PM To: Microscopy-at-msa.microscopy.com
We are comparing dye sub printers versus the Fuji Pictrography PG4500. The price differential is a staggering $22,000 versus $6,500, respectively. It is my understanding that both produce good quality, gray-scale prints (versus dithered, inkjet prints). Why the price differential? I would welcome any comments, user experiences, etc. Does anyone know the average cost per 8.5 x 11in print?
Thank you. -- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Email: bozzola-at-siu.edu ##############################################################
I have updated the security model and the software for the Listserver this morning , and need to perform a full mailing test to all subscribers to confirm functionality. Basically I believe that I have patched a few software holes that spammers have discovered.
I will be monitoring the system for problems the rest of the day. Hopefully this test will run fine and there will be minimal interruptions.
Should you encounter problems please contact me off-line (zaluzec-at-microscopy.com).
Cheers
Nestor Your Friendly Neighborhood SysOp
Sunday July 3, 2005 11:20 AM CST -----------------------------
Two similar models of this monitor were available: IBM T221 and Viewsonic VP2290B, both using IBM display panel 22" diagonal, 3840 x 2400 pixels, 204 dpi. Both discontinued. IBM had intentions of replacing T221 with new model as early as March 2005. I don't know whether they did yet.
We purchased several such monitors, IBM and Viewsonic, for TEM camera systems. Both models are similar in performance, except Viewsonic is a bit slower in full resolution mode, which makes no difference for static images. It's hard for a PC to run 9.2 MP frames at over 20 FPS refresh rate anyway. In fact, it is better to run 1600 x 1200 desktop most of the time and switch to 3840 x 2400 for high resolution viewing. 1600 x 1200 keeps fonts and icons size comfortable, and refresh rate is fast. The display in 3840 x 2400 mode is stunning- like looking through a window on a sunny day. You can take an eye loupe to the screen and see further detail in the image.
If you will be able to find refurbished or second hand T221 - IBM will honor original 3 year factory warranty as long as monitor is not physically damaged. One of our IBM monitors was refurbished, with a screen defect. IBM replaced the monitor. Consult with IBM regarding the warranty, before buying used monitor. Have monitor serial number when calling IBM. These monitors are still available, mostly on e-bay, and through some internet outlets. Could be even new in box, but not from a regular source.
Is anybody at IBM reading this message? Please comment on a future availability of equal or better display monitor.
See T221 in EM application at www-1.ibm.com/industries/healthcare/doc/content/bin/ls_t221_enhancing_electr on_1.pdf
Vitaly Feingold SIA 2773 Heath Lane Duluth, GA 30096 Ph. 770-232-7785 Fax 770-232-1791 www.sia-cam.com
----- Original Message ----- } From: "Mardinly, John" {john.mardinly-at-intel.com} To: "Don Chernoff at ASM" {donc-at-asmicro.com} ; "Microscopy List" {microscopy-at-microscopy.com} Sent: Saturday, July 02, 2005 3:53 AM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (amr2w-at-virginia.edu) from http://microscopy.com/MLFormMail.html on Thursday, June 30, 2005 at 12:58:42 ---------------------------------------------------------------------------
Email: amr2w-at-virginia.edu Name: Andrew Roelant
Organization: UVa
Title-Subject: [Microscopy] [Filtered] 3-D SEM reconstruction
Question: Hi all, I know there are software programs that do 3-D images from SEM using stereo pairs, however, what if you have a more complex shaped object? I hear that there has been new software that you can use a series of tilts like TEM tomography to obtain a better reconstruction. Does anybody know about this? Any information would be very much appreciated. Thanks! -Andrew Roelant
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (srandol3-at-utk.edu) from http://www.microscopy.com/MLFormMail.html on Sunday, July 3, 2005 at 19:26:02 ---------------------------------------------------------------------------
Email: srandol3-at-utk.edu Name: Steven Randolph
Organization: university of tennessee
Title-Subject: [Microscopy] [Filtered] MListserver: Area analysis scanning
Question: Hello all,
I have a question that is really more engineering-related than imaging. My question is regarding some of the various scanning modes on Hitachi SEMs such as the S4300, S4700, and S3500N. I need to know some of the specifics about how scanning takes place in area analysis mode and reduced screen mode. In area analysis, is the beam blanked in the region outside the box, or is the pixel size reduced to accomodate the box size? I guess what I'm trying to find out is the dwell time in area analysis and reduced screen mode. I seem to recall that there is a finite settle time associated with scanning in some modes. Anyways, any input you could provide would be much appreciated!
We have a Philips SEM 515 equipped with EDX spectrometer. The spectrometer is EDAX PV 9800 system with UTW detector (freshly repaired) and control unit with its specialized computer (both hardware and software). The computer is just dying and most probably can not be repaired. I wonder if anybody tried to "upgrade" this particular system to PC compatible version. Is it possible at reasonable cost?
Regards,
Leszek
Leszek Kepinski Institute of Low Temperature and Structure Research, Polish Academy of Sciences, P.O.Box 1410, 50-950 Wroclaw, Poland e-mail: L.Kepinski-at-int.pan.wroc.pl
Well, it was wishful thinking that I could adequitely solve the problem in the first attempt. After many hours of off-line testing (and too few pints for a holiday weekend). I need to run yet another full system delivery test. You may notice a few changes in the configuration of the Email headers in this version.
The previous test basically worked, but it took far too many hours to process & verify all the addresses. I've taken a whole different aproach with this modification (after having looked at the results over night).
No need to reply to this message. Again if you have problems posting with this new version of the filters implements, then please contact me off-line. (zaluzec-at-microscopy.com)
Nestor Your Friendly Neighborhood SysOp.
Monday - July 4th, 2005
------------------------------Original Headers------------------------------ 16, 11 -- From zaluzec-at-microscopy.com Mon Jul 4 13:54:57 2005 16, 11 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) 16, 11 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j64Ist7n013713 16, 11 -- for {microscopy-at-microscopy.com} ; Mon, 4 Jul 2005 13:54:57 -0500 16, 11 -- Mime-Version: 1.0 16, 11 -- Message-Id: {p06110449beef32f13eb5-at-[206.69.208.22]} 16, 11 -- Date: Mon, 4 Jul 2005 13:54:54 -0500 16, 11 -- To: microscopy-at-microscopy.com 16, 11 -- From: "Nestor J. Zaluzec" {zaluzec-at-microscopy.com} 16, 11 -- Subject: Administrivia: Sorry - another full system test 16, 11 -- Content-Type: text/plain; charset="us-ascii" ------------------------------------------------------------------------
The next in the FOM conference series will take place in Perth, (Western) Australia from Sunday April 9th to Wednesday April 12, 2006. Please visit for details www.FocusOnMicroscopy.org
Focus on Microscopy 2006 is the continuation of a successful conference series presenting the latest innovations in optical microscopy and its applications in biology, medicine, material science, and information storage. 3D optical imaging and related theory are important subjects for the conference. The series is as relevant now as at any time in its history as the scientific and engineering communities strive to meet the needs of a surging life sciences sector as well as respond to the sustained pressure on miniaturisation in lithography and data storage.
Also a technical exhibition will be part of the conference. In Jena well over 30 companies participated including the major microscopy companies, see FOM2005 Sponsors & Exhibitors www.focusonmicroscopy.org/2005/sponsors.html .
The 2006 meeting will be held in Perth on Australia's western seaboard. The conference will be held in the nearby port city of Fremantle, at the scenic Esplanade hotel, close to the boat harbour and Perth' famous beaches. Perth's relaxed and outdoor lifestyle should prove an ideal setting for a stimulating and enjoyable meeting - see you there!
Local organising committee: Stephen Cody Central Resource for Advanced Microscopy, Ludwig Institute for Cancer Research, Melbourne
Guy Cox Electron Microscope Unit, University of Sydney
Ewa Goldys Division of Information and Communication Sciences, Macquarie University, Sydney
Brendan Griffin Centre for Microscopy and Microanalysis, University of Western Australia, Perth
Miranda Grounds School of Anatomy and Human Biology, University of Western Australia, Perth
Ian Harper School of Biomedical Sciences, Monash University, Melbourne
David Jans Department of Biochemistry and Molecular Biology, Monash University, Melbourne
Min Gu Centre for Microphotonics, Swinburne University, Melbourne
John Kuo Centre for Microscopy and Microanalysis, University of Western Australia, Perth
Keith Nugent School of Physics, University of Melbourne
Paul Rigby Biomedical Imaging and Analysis Facility, University of Western Australia, Perth
Alpha Yap Institute for Biomolecular Science, University of Queensland, Brisbane
Conference topics include: * Confocal and multiphoton-excitation microscopies * Novel illumination and detection strategies - selectiveplane, extended depth of focus, 4pi, structured illumination * Fluorescence - new labels, fluorescent proteins, quantum dots, single molecule, excitation-emission spectroscopy * Time-resolved fluorescence - FRET, FRAP, FLIM, FCS * Coherent non-linear microscopies - SHG, THG, SFG, CARS * Scattering processes: Raman, light scattering spectroscopy, second harmonic * Multi-dimensional imaging * Sub-wavelength resolution - near field microscopy, total internal reflection * Laser manipulation, ablation and microdissection, photoactivation * Magnetic resonance and X-ray microscopy * Image processing and visualisation * Live cell and tissue imaging * Whole tissue imaging - optical coherence tomography, endoscopy, whole animal fluorescence * New tools in genomics, proteomics, phenomics, cytometry * Lithography and data storage
To stay informed on the program, registration and abstract submission for the conference please leave your E-mail address here www.focusonmicroscopy.org/forms/subscr_notification.html .
On behalf of the FocusOnMicroscopy society, * David Sampson, University of Western Australia * Fred Brakenhoff, University of Amsterdam, The Netherlands
Stephen H. Cody
Microscopy Manager Central Resource for Advanced Microscopy Ludwig Institute For Cancer Research PO Box 2008 Royal Melbourne Hospital Parkville Victoria 3050 Australia Tel: 61 3 9341 3155 Fax: 61 3 9341 3104 email: stephen.cody-at-ludwig.edu.au www.ludwig.edu.au/labs/confocal.html www.ludwig.edu.au/confocal
------------------------------Original Headers------------------------------ 23, 23 -- From Stephen.Cody-at-ludwig.edu.au Mon Jul 4 19:10:35 2005 23, 23 -- Received: from CL380EVS-1.ludwig.edu.au (cl380-1.ludwig.edu.au [128.250.250.24]) 23, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j650AXXm030562 23, 23 -- for {microscopy-at-microscopy.com} ; Mon, 4 Jul 2005 19:10:34 -0500 23, 23 -- Received: from exchange.ludwig.edu.au ([172.16.2.22]) by CL380EVS-1.ludwig.edu.au with Microsoft SMTPSVC(5.0.2195.6713); 23, 23 -- Tue, 5 Jul 2005 10:10:33 +1000 23, 23 -- Content-Class: urn:content-classes:message 23, 23 -- MIME-Version: 1.0 23, 23 -- Content-Type: text/plain; 23, 23 -- charset="iso-8859-1" 23, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 23, 23 -- Subject: FOM 2006 23, 23 -- Date: Tue, 5 Jul 2005 10:10:33 +1000 23, 23 -- Message-ID: {660936C3FAE0FB47BC49BB8B60FA14EC148BFE-at-exchange.ludwig.edu.au} 23, 23 -- X-MS-Has-Attach: 23, 23 -- X-MS-TNEF-Correlator: 23, 23 -- Thread-Topic: FOM 2006 23, 23 -- thread-index: AcWA9fXZB44rSoAySq+8wRoucYuX8Q== 23, 23 -- From: "Stephen Cody" {Stephen.Cody-at-ludwig.edu.au} 23, 23 -- To: {microscopy-at-microscopy.com} 23, 23 -- X-OriginalArrivalTime: 05 Jul 2005 00:10:33.0257 (UTC) FILETIME=[F6019990:01C580F5] 23, 23 -- Content-Transfer-Encoding: 8bit 23, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id j650AXXm030562
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Postdoctoral Position: Computer Vision Algorithms for Studying Biological Growth and Motility
A postdoctoral position is available to join a research project in the quantification of deformable motion in biology, with emphasis on growth and cell motility. The position is supported by an NIH-funded collaboration between Tobias I. Baskin (a biologist at Umass Amherst) and K. Palaniappan (a computer scientist at University of Missouri, Columbia). Baskin and Palaniappan have developed new software for quantifying the spatial distribution of velocity within a growing plant organ (a root). The software is called RootflowRT and the biological application is described by van der Weele et al (2003 Plant Physiology, 32:1138-1148). The software implements a novel algorithm for quantifying deformable motion that combines structure-tensor and robust-matching approaches. The project is to enhance and validate RootFlowRT, apply software engineering principles to the current code base, explore new computational algorithms, and extend the robust-tensor approach to other kinds of biological objects, in particular motile animal cells and embryos. The open position is at Amherst and involves imaging different kinds of biological object as well as enhancing the software. Applicants should have experience in some area of image processing, good programming skills, and, preferably, experience in biology.
Those interested in the position should contact Dr Baskin (email: baskin-at-bio.umass.edu), and can find further information from his web page: http://www.bio.umass.edu/biology/baskin/ and the page for RootflowRT http://meru.rnet.missouri.edu/mvl/bio_motion.
I encourage applications from anyone regardless of skin color, religion, sex, sexual orientation, or nationality.
Hi, Is anyone aware of a method using DiI for staining cells growing in a monolayer thickness, targeting the cell membrane, to obtain enough contrast to yield a binary image of the cells representing their shape/morphology?
I'm growing multi-potent cells in 24-well plates and would like to quantify their shape (and changes in shape) as they differentiate with time.
I've looked in the literature, and I've found lots of examples of DiI being used to label neurons in live tissue. However, these papers don't focus so much on quantitative morphological measurements, but more on what the neurons are ennervating. I'd like a simple method for cells growing or fixed in a plate or on a slide, for quantifying shape.
I am looking for companies or individuals providing repair services for TEMs and SEMs in the New England area. Does anyone know of a comprehensive list of contacts? Any recommendations?
Any providers or others with relevant information are welcome to contact me at the phone number listed below, or by e-mail. Thanks.
Marie
Dr. Marie E. Cantino Director, Electron Microscopy Laboratory Associate Professor of Physiology and Neurobiology University of Connecticut Unit 3242 Storrs, CT 06269-3242 Phone: 860-486-3588 Fax: 860-486-6369
==============================Original Headers============================== 5, 20 -- From marie.cantino-at-uconn.edu Wed Jul 6 13:39:00 2005 5, 20 -- Received: from mail2.uits.uconn.edu (mail2.uits.uconn.edu [137.99.25.204]) 5, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j66IcxI8001091 5, 20 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 6 Jul 2005 13:39:00 -0500 5, 20 -- Received: from [137.99.47.197] (d47h197.public.uconn.edu [137.99.47.197]) 5, 20 -- by mail2.uits.uconn.edu (8.11.6/8.11.6) with ESMTP id j66IcmO19281 5, 20 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 6 Jul 2005 14:38:48 -0400 5, 20 -- Mime-Version: 1.0 (Apple Message framework v622) 5, 20 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 5, 20 -- Message-Id: {84b91857580009a34dab5aecf6fd70a7-at-uconn.edu} 5, 20 -- Content-Transfer-Encoding: 7bit 5, 20 -- Reply-To: Cantino Marie {marie.cantino-at-uconn.edu} 5, 20 -- From: Marie Cantino {marie.cantino-at-uconn.edu} 5, 20 -- Subject: EM service 5, 20 -- Date: Wed, 6 Jul 2005 14:44:15 -0400 5, 20 -- To: MSA Listserver {Microscopy-at-msa.microscopy.com} 5, 20 -- X-Mailer: Apple Mail (2.622) 5, 20 -- X-UConn-MailScanner-Information: Contact UConn Help Desk 860-486-4357 for more information. 5, 20 -- X-UConn-MailScanner: Found to be clean 5, 20 -- X-UConn-MailScanner-SpamCheck: ==============================End of - Headers==============================
Ken Converse Quality Images Delta PA (717) 456-5491
He participates in this newsgroup.
Stu Smalinskas Metallurgist SKF Plymouth, Michigan
--- marie.cantino-at-uconn.edu wrote:
} } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } I am looking for companies or individuals providing } repair services for } TEMs and SEMs in the New England area. Does anyone } know of a } comprehensive list of contacts? Any } recommendations? } } Any providers or others with relevant information } are welcome to } contact me at the phone number listed below, or by } e-mail. Thanks. } } Marie } } Dr. Marie E. Cantino } Director, Electron Microscopy Laboratory } Associate Professor of Physiology and Neurobiology } University of Connecticut Unit 3242 } Storrs, CT 06269-3242 } Phone: 860-486-3588 } Fax: 860-486-6369 }
____________________________________________________ Sell on Yahoo! Auctions – no fees. Bid on great items. http://auctions.yahoo.com/
I'm interested in hearing how long people keep the Uranyl Acetate stain that they use in grid staining, both the stock and working solution. Currently we are using a 2% solution in 50% methanol.
How long do you think one could say that they remain fresh after making up? Presently our lab keeps the stock solution for about a month, but I never really had any literature to know one way or the other if I could keep it long.
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==============================Original Headers============================== 5, 19 -- From GBurgess-at-exchange.hsc.mb.ca Wed Jul 6 14:20:58 2005 5, 19 -- Received: from hscxntmx0003.hsc.mb.ca (hscxntmx0003.hsc.mb.ca [142.233.100.122]) 5, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j66JKw26016949 5, 19 -- for {Microscopy-at-microscopy.com} ; Wed, 6 Jul 2005 14:20:58 -0500 5, 19 -- Received: from mudslide.hsc.mb.ca (unverified [172.16.6.136]) by 5, 19 -- hscxntmx0003.hsc.mb.ca(Vircom SMTPRS 4.0.346.0) with ESMTP id 5, 19 -- {B0013187032-at-hscxntmx0003.hsc.mb.ca} for {Microscopy-at-microscopy.com} ;Wed, 5, 19 -- 6 Jul 2005 14:23:29 -0500 5, 19 -- Received: by mudslide.hsc.mb.ca with Internet Mail Service (5.5.2653.19)id 5, 19 -- {3F0H7VDL} ; Wed, 6 Jul 2005 14:20:52 -0500 5, 19 -- Message-ID: {00A937989100304A83A058F6C45873FF32A241-at-hscxntmx0005} 5, 19 -- Date: Wed, 6 Jul 2005 14:19:35 -0500 5, 19 -- From: Garry Burgess {GBurgess-at-exchange.hsc.mb.ca} 5, 19 -- Subject: Uranyl Acetate Shelf Life 5, 19 -- To: {Microscopy-at-microscopy.com} 5, 19 -- X-Mailer: Internet Mail Service (5.5.2653.19) 5, 19 -- MIME-Version: 1.0 5, 19 -- Content-Type: text/plain; 5, 19 -- charset="iso-8859-1" ==============================End of - Headers==============================
We've been using multiple LCD monitors lately to increase our windows desktop size. However, in doing so I have noted that Matrox does offer a high-resolution graphics card (3840x2400) { http://www.matrox.com/mga/workstation/3dws/products/special/hr25 6.cfm } and they point to three monitors for this card something refed to as 9 MP (assume 9 mega pixel ? maybe) monitors (Viewsonic, IBM T221, and Iiayam)
NVidia Quadro FX series cards also goes to 3840x2400
ATI as a few cards which will deliver 3840 x 2400 (using multiple monitors)
IBM lists a T221 as their 9.2 megapixel monitor - but I can not find its avavilablity.
A company called Planar makes a 5-megapixel greyscale monitor. Dome C5i
www.tridentmicrosystems.co.uk lists a 28.1” 2k x 2k TFT LCD monitor is designed for traffic management.
NEC lists 2048 x 1536 as their highest resolution monitor. (But does include 10-bit greyscale as well).
Viewsonic offers a number of "Thin Edge" monitors designed to used together including stands designed to hold 2,3 or 4 monitors at once.
There is a company www.9xmedia.com that offers multimonitor solutions.
(Note: I do not sell montiors, or own stock / interests in any monitor graphics company. :-)
} ------------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } Don; } I tried to purchase an IBM T-220 9 megapixel display, but was } told by IBM that they had killed the product. This was the only display } ever marketed that could display a 2K X 2K camera image showing all of } the pixels on one screen. We still have two screens on our TEM, but need } to either display the images at half resolution, or zoom to display only } part of the image. } } John Mardinly } } -----Original Message----- } } From: Don Chernoff at ASM [mailto:donc-at-asmicro.com] } Sent: Friday, July 01, 2005 9:05 AM } To: Microscopy List } Subject: [Microscopy] Image review: Hard copy vs. PC screen } } } } ------------------------------------------------------------------------ } ------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ } ------- } } This post is tangential to the dye sub vs ink jet discussion. } When one needs to compare 2 or more images simultaneously, prints can } easily } be laid out on a table, but this is not so easy with on-screen display } of } digital images. One thing that we have done in our lab is to install a } second monitor at some workstations and increase the Windows desktop } size so } that it spans the two monitors. This is satisfactory for AFM images } where } the basic pixel count is 512x512 but may not be so good with other } formats. } I am curious to learn what other people do. } } When you give your customers digital images only, do you feel there is a } risk they could miss an important comparison? } } regards, } Don Chernoff } ================================== } Advanced Surface Microscopy, Inc. E-Mail: donc-at-asmicro.com } 3250 N. Post Rd., Ste. 120 Voice: 317-895-5630 } INDIANAPOLIS IN 46226 USA Toll free: 800-374-8557 (in USA & } Canada) } web: http://www.asmicro.com Fax: 317-895-5652 } [business activities: analytical services in AFM, AFM probes, } consulting, } training, } calibration and test specimens, calibration and measurement software, } used NanoScope equipment.] } } } }
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Director 350 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
"WE ARE MICROSOFT. RESISTANCE IS FUTILE. YOU WILL BE ASSIMILATED."
==============================Original Headers============================== 21, 25 -- From edelmare-at-muohio.edu Wed Jul 6 16:05:08 2005 21, 25 -- Received: from mulnx11.mcs.muohio.edu (mulnx11.mcs.muohio.edu [134.53.6.66]) 21, 25 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j66L582x025686 21, 25 -- for {microscopy-at-Microscopy.com} ; Wed, 6 Jul 2005 16:05:08 -0500 21, 25 -- Received: from muw2k04 (muw2k04.mcs.muohio.edu [134.53.6.18]) 21, 25 -- by mulnx11.mcs.muohio.edu (Switch-3.1.6/Switch-3.1.6) with SMTP id j66L53TN032744 21, 25 -- for {microscopy-at-Microscopy.com} ; Wed, 6 Jul 2005 17:05:03 -0400 21, 25 -- Received: From mcsaix06.mcs.muohio.edu ([134.53.253.28]) by muw2k04 (WebShield SMTP v4.5 MR1a P0803.345); 21, 25 -- id 1120683626359; Wed, 6 Jul 2005 17:00:26 -0400 21, 25 -- Received: from emf03 ([134.53.14.119]) 21, 25 -- by mcsaix06.mcs.muohio.edu (Switch-3.1.6/Switch-3.1.6) with ESMTP id j66L4wMJ013798; 21, 25 -- Wed, 6 Jul 2005 17:04:58 -0400 21, 25 -- From: "Richard E. Edelmann" {edelmare-at-muohio.edu} 21, 25 -- To: "Mardinly, John" {john.mardinly-at-intel.com} , microscopy-at-Microscopy.com 21, 25 -- Date: Wed, 6 Jul 2005 17:05:02 -0400 21, 25 -- MIME-Version: 1.0 21, 25 -- Content-type: text/plain; charset=ISO-8859-1 21, 25 -- Subject: Re: [Microscopy] RE: High Res Monitors 21, 25 -- Message-ID: {42CC0F3E.21724.1D578A6-at-localhost} 21, 25 -- Priority: normal 21, 25 -- In-reply-to: {9E1ED6A623D8CB44B4866ACF20ECDB2B06C779AA-at-scsmsx403.amr.corp.intel.com} 21, 25 -- X-mailer: Pegasus Mail for Win32 (v3.12c) 21, 25 -- X-Scanned-By: MIMEDefang 2.45 21, 25 -- Content-Transfer-Encoding: 8bit 21, 25 -- X-MIME-Autoconverted: from Quoted-printable to 8bit by ns.microscopy.com id j66L582x025686 ==============================End of - Headers==============================
We have an old Zeiss EM10C, which we try to put back to the service. It come with dual channel Roentgen meter board, MOSFET. Unfortunately we do not have a circuit diagram, which will allow us to fix a problem on the board. Some one has a copy of diagram of that board?
Thanks, Vlad
==============================Original Headers============================== 5, 19 -- From uti-at-direcpc.com Thu Jul 7 06:41:13 2005 5, 19 -- Received: from a34-mta01.direcway.com (a34-mta01.direcpc.com [66.82.4.90]) 5, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j67BfD5X031659 5, 19 -- for {microscopy-at-microscopy.com} ; Thu, 7 Jul 2005 06:41:13 -0500 5, 19 -- Received: from ringo.direcpc.com (dpclt034012.direcpc.com [64.157.34.12]) 5, 19 -- by a34-mta01.direcway.com 5, 19 -- (iPlanet Messaging Server 5.2 HotFix 2.05 (built Mar 3 2005)) 5, 19 -- with ESMTP id {0IJ900DYB9S9LS-at-a34-mta01.direcway.com} for 5, 19 -- microscopy-at-microscopy.com; Thu, 07 Jul 2005 07:41:09 -0400 (EDT) 5, 19 -- Date: Thu, 07 Jul 2005 07:41:48 -0400 5, 19 -- From: UTI {uti-at-direcpc.com} 5, 19 -- Subject: Zeiss EM10C - Roentgen meter board 5, 19 -- X-Sender: uti-at-pop3.direcpc.com 5, 19 -- To: microscopy-at-microscopy.com 5, 19 -- Message-id: {5.1.0.14.2.20050707073518.01e65a30-at-pop3.direcpc.com} 5, 19 -- MIME-version: 1.0 5, 19 -- X-Mailer: QUALCOMM Windows Eudora Version 5.1 5, 19 -- Content-type: text/plain; charset=us-ascii; format=flowed 5, 19 -- Content-transfer-encoding: 7BIT ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jmjenks-at-pacbell.net) from http://microscopy.com/MLFormMail.html on Thursday, July 7, 2005 at 09:31:04 ---------------------------------------------------------------------------
Email: jmjenks-at-pacbell.net Name: Jeff Jenks
Organization: micronetpartners.com
Title-Subject: [Filtered] Senior EPMA (Electron Probe Micro Analysis) Scientist position
Silicon Valley based semiconductor equipment company seeks Senior EPMA Scientist as follows:
Mission and Duties:
- Actively participate in the development, testing and evaluation of new metrology solutions for semiconductor processing from the concept stage to prototype testing.
- Provide a broad practical and theoretical knowledge base of electron spectroscopy, X-ray optics, and electron probe micro analysis (EPMA) to assess analytical capabilities, application areas and metrology opportunities of various analysis techniques.
- Work with a team of scientists and engineers in system development, prototype testing, and design of experiments.
- Plan, perform and evaluate hardware and application feasibility studies on test setups and complete prototype systems.
Responsibilities:
- Independently perform and evaluate experiments using prototype systems
- Design experiments for EPMA concept feasibility
- Build and modify experimental test systems using hands-on skills
- Provide and maintain up-to-date knowledge of surface characterization techniques
- Model and predict electron and X-ray intensities due to particle-material interaction
- Develop, test, and evaluate measurement protocols
- Actively participate in system development and subsystem designs
Background and Education:
- MS or Ph.D. in material science, physics or related field is required; an emphasis in measurement or instrumentation is preferred
- Broad knowledge in material analysis
- Strong practical experience in electron probe analysis, preferably WDX Wavelength- Dispersive X-ray Spectroscopy or EPMA or equivalent
- Experience in semiconductor material analysis is preferred but not required
- Intimate knowledge of data reduction methods, statistical analysis and principles of particle-material interaction
- Knowledge of X-ray optics and/or X-ray optical design is desirable
- Ability to work independently and within a team of scientists and engineers
- Minimum of 2 years of practical experience in a combination of the above areas
Company offers competitive salary, stock options, benefits, and relocation assistance in a collegial work environment. Company is privately held and VC-funded with outstanding prospects for stock price appreciation in the future.
For immediate consideration please contact Jeff Jenks with your resume or CV at Jeff-at-micronetpartners.com. Or you can call Jeff at 408-850-7271 for additional details. No visa sponsorship is available. For additional openings please view www.micronetpartners.com/jobs.html.
Sorry, I seem to have scrambled the subject line of the last few postings with my latest version of the filter code. Please do not critique the person posting it was my fault.
Nestor Your Friendly Neighborhood SysOp
-- =========================================== Dr. Nestor J. Zaluzec Argonne National Lab Materials Science Division/Bldg 212 9700 S. Cass Ave Argonne, Illinois 60439 USA Tel: 630-252-7901, Fax: 630-252-4798 Email: Zaluzec-at-aaem.amc.anl.gov =========================================== TPMLab: http://tpm.amc.anl.gov NEESLab: http://neestpm.mcs.anl.gov MMSite: http://www.amc.anl.gov ===========================================
The box said ... "This program requires Win 95/98/NT or better..." So I bought a G3 Mac !
===========================================
==============================Original Headers============================== 7, 14 -- From zaluzec-at-aaem.amc.anl.gov Thu Jul 7 10:45:31 2005 7, 14 -- Received: from aaem.amc.anl.gov (aaem.amc.anl.gov [146.139.72.3]) 7, 14 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j67FjVae018382 7, 14 -- for {microscopy-at-microscopy.com} ; Thu, 7 Jul 2005 10:45:31 -0500 7, 14 -- Received: from [146.139.72.105] (aem105.amc.anl.gov [146.139.72.105]) 7, 14 -- by aaem.amc.anl.gov (8.12.11/8.12.10) with ESMTP id j67FjT3a027583 7, 14 -- for {microscopy-at-microscopy.com} ; Thu, 7 Jul 2005 10:45:31 -0500 7, 14 -- Mime-Version: 1.0 7, 14 -- Message-Id: {p0611040cbef2fe9749d0-at-[146.139.72.105]} 7, 14 -- Date: Thu, 7 Jul 2005 10:45:29 -0500 7, 14 -- To: microscopy-at-microscopy.com 7, 14 -- From: "Nestor J. Zaluzec" {zaluzec-at-aaem.amc.anl.gov} 7, 14 -- Subject: Administrivia: Subject Line scrambled...sorry 7, 14 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
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Colleagues
Sorry, I seem to have scrambled the subject line of the last few postings with my latest version of the filter code. Please do not critique the person posting it was my fault.
Nestor Your Friendly Neighborhood SysOp
-- =========================================== Dr. Nestor J. Zaluzec Argonne National Lab Materials Science Division/Bldg 212 9700 S. Cass Ave Argonne, Illinois 60439 USA Tel: 630-252-7901, Fax: 630-252-4798 Email: Zaluzec-at-aaem.amc.anl.gov =========================================== TPMLab: http://tpm.amc.anl.gov NEESLab: http://neestpm.mcs.anl.gov MMSite: http://www.amc.anl.gov ===========================================
The box said ... "This program requires Win 95/98/NT or better..." So I bought a G3 Mac !
===========================================
==============================Original Headers============================== 13, 14 -- From zaluzec-at-aaem.amc.anl.gov Thu Jul 7 10:48:26 2005 13, 14 -- Received: from aaem.amc.anl.gov (aaem.amc.anl.gov [146.139.72.3]) 13, 14 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j67FmPWm018432 13, 14 -- for {microscopy-at-microscopy.com} ; Thu, 7 Jul 2005 10:48:25 -0500 13, 14 -- Received: from [146.139.72.105] (aem105.amc.anl.gov [146.139.72.105]) 13, 14 -- by aaem.amc.anl.gov (8.12.11/8.12.10) with ESMTP id j67FmO1b027590 13, 14 -- for {microscopy-at-microscopy.com} ; Thu, 7 Jul 2005 10:48:24 -0500 13, 14 -- Mime-Version: 1.0 13, 14 -- Message-Id: {p0611040dbef2ff1a687b-at-[146.139.72.105]} 13, 14 -- Date: Thu, 7 Jul 2005 10:48:23 -0500 13, 14 -- To: microscopy-at-microscopy.com 13, 14 -- From: "Nestor J. Zaluzec" {zaluzec-at-aaem.amc.anl.gov} 13, 14 -- Subject: [Microscopy] Administrivia: Subject Line scrambled...sorry 13, 14 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jmjenks-at-pacbell.net) from http://microscopy.com/MLFormMail.html on Thursday, July 7, 2005 at 09:31:04 ---------------------------------------------------------------------------
Email: jmjenks-at-pacbell.net Name: Jeff Jenks
Organization: micronetpartners.com
Title-Subject: [Filtered] Senior EPMA (Electron Probe Micro Analysis) Scientist position
Silicon Valley based semiconductor equipment company seeks Senior EPMA Scientist as follows:
Mission and Duties:
- Actively participate in the development, testing and evaluation of new metrology solutions for semiconductor processing from the concept stage to prototype testing.
- Provide a broad practical and theoretical knowledge base of electron spectroscopy, X-ray optics, and electron probe micro analysis (EPMA) to assess analytical capabilities, application areas and metrology opportunities of various analysis techniques.
- Work with a team of scientists and engineers in system development, prototype testing, and design of experiments.
- Plan, perform and evaluate hardware and application feasibility studies on test setups and complete prototype systems.
Responsibilities:
- Independently perform and evaluate experiments using prototype systems
- Design experiments for EPMA concept feasibility
- Build and modify experimental test systems using hands-on skills
- Provide and maintain up-to-date knowledge of surface characterization techniques
- Model and predict electron and X-ray intensities due to particle-material interaction
- Develop, test, and evaluate measurement protocols
- Actively participate in system development and subsystem designs
Background and Education:
- MS or Ph.D. in material science, physics or related field is required; an emphasis in measurement or instrumentation is preferred
- Broad knowledge in material analysis
- Strong practical experience in electron probe analysis, preferably WDX Wavelength- Dispersive X-ray Spectroscopy or EPMA or equivalent
- Experience in semiconductor material analysis is preferred but not required
- Intimate knowledge of data reduction methods, statistical analysis and principles of particle-material interaction
- Knowledge of X-ray optics and/or X-ray optical design is desirable
- Ability to work independently and within a team of scientists and engineers
- Minimum of 2 years of practical experience in a combination of the above areas
Company offers competitive salary, stock options, benefits, and relocation assistance in a collegial work environment. Company is privately held and VC-funded with outstanding prospects for stock price appreciation in the future.
For immediate consideration please contact Jeff Jenks with your resume or CV at Jeff-at-micronetpartners.com. Or you can call Jeff at 408-850-7271 for additional details. No visa sponsorship is available. For additional openings please view www.micronetpartners.com/jobs.html.
Sorry, I seem to have scrambled the subject line of the last few postings with my latest version of the filter code. Please do not critique the person posting it was my fault.
I have reposted at least one of the scrambled messages.
Nestor Your Friendly Neighborhood SysOp
==============================Original Headers============================== 8, 14 -- From zaluzec-at-aaem.amc.anl.gov Thu Jul 7 10:51:16 2005 8, 14 -- Received: from aaem.amc.anl.gov (aaem.amc.anl.gov [146.139.72.3]) 8, 14 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j67FpEIB026871 8, 14 -- for {microscopy-at-microscopy.com} ; Thu, 7 Jul 2005 10:51:15 -0500 8, 14 -- Received: from [146.139.72.105] (aem105.amc.anl.gov [146.139.72.105]) 8, 14 -- by aaem.amc.anl.gov (8.12.11/8.12.10) with ESMTP id j67FpEge027604 8, 14 -- for {microscopy-at-microscopy.com} ; Thu, 7 Jul 2005 10:51:14 -0500 8, 14 -- Mime-Version: 1.0 8, 14 -- Message-Id: {p0611040ebef2ffbc8e92-at-[146.139.72.105]} 8, 14 -- Date: Thu, 7 Jul 2005 10:51:13 -0500 8, 14 -- To: microscopy-at-microscopy.com 8, 14 -- From: "Nestor J. Zaluzec" {zaluzec-at-aaem.amc.anl.gov} 8, 14 -- Subject: [Microscopy] Administrivia: Subject Line scrambled...sorry 8, 14 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
Greetings from Baltimore. I have a Zeiss 10 CA. Our past service company Pesto Inc. longer is in business. Does anyone know of a person or company that works on Zeiss TEMs in my area? -- Chere Petty, M.S. Manager, Keith R. Porter Imaging Facility Department of Biological Sciences University of Maryland Baltimore County (UMBC) 1000 Hilltop Circle Baltimore, MD 21250 Phone: 410-455-2296 Fax: 410-455-3875
==============================Original Headers============================== 1, 21 -- From cpetty1-at-umbc.edu Thu Jul 7 11:58:28 2005 1, 21 -- Received: from mx1out.umbc.edu (mx1out.umbc.edu [130.85.25.10]) 1, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j67GwRfI010236 1, 21 -- for {Microscopy-at-Microscopy.com} ; Thu, 7 Jul 2005 11:58:27 -0500 1, 21 -- Received: from [130.85.116.27] (biosci83pc-01.biosci.umbc.edu [130.85.116.27]) 1, 21 -- by mx1out.umbc.edu (8.12.10/8.12.10/UMBC-Central 1.1.2.1 mxout 1.2.2.3) with ESMTP id j67GwP7T009436 1, 21 -- for {Microscopy-at-Microscopy.com} ; Thu, 7 Jul 2005 12:58:26 -0400 (EDT) 1, 21 -- Message-ID: {42CD5F2B.1000908-at-umbc.edu} 1, 21 -- Date: Thu, 07 Jul 2005 12:58:19 -0400 1, 21 -- From: Chere Petty {cpetty1-at-umbc.edu} 1, 21 -- Reply-To: cpetty1-at-umbc.edu 1, 21 -- Organization: Biological Sciences - UMBC 1, 21 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.5) Gecko/20041217 1, 21 -- X-Accept-Language: en-us, en 1, 21 -- MIME-Version: 1.0 1, 21 -- To: "Microscopy-at-Microscopy.com" {Microscopy-at-Microscopy.com} 1, 21 -- Subject: TEM local service 1, 21 -- Content-Type: text/plain; charset=us-ascii; format=flowed 1, 21 -- Content-Transfer-Encoding: 7bit 1, 21 -- X-AvMilter-Key: 1120755807:ba5319d8d857ad1aef62579cf7f2f12f 1, 21 -- X-Avmilter: Message Skipped, too small ==============================End of - Headers==============================
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Dear group,
We have an old Zeiss EM10C, which we try to put back to the service. It come with dual channel Roentgen meter board, MOSFET. Unfortunately we do not have a circuit diagram, which will allow us to fix a problem on the board. Some one has a copy of diagram of that board?
Thanks, Vlad
==============================Original Headers============================== 9, 15 -- From zaluzec-at-microscopy.com Thu Jul 7 12:06:12 2005 9, 15 -- Received: from aaem.amc.anl.gov (aaem.amc.anl.gov [146.139.72.3]) 9, 15 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j67H6BRu018004 9, 15 -- for {microscopy-at-microscopy.com} ; Thu, 7 Jul 2005 12:06:11 -0500 9, 15 -- Received: from [146.139.72.105] (aem105.amc.anl.gov [146.139.72.105]) 9, 15 -- by aaem.amc.anl.gov (8.12.11/8.12.10) with ESMTP id j67H6Ar2027721 9, 15 -- for {microscopy-at-microscopy.com} ; Thu, 7 Jul 2005 12:06:11 -0500 9, 15 -- Mime-Version: 1.0 9, 15 -- X-Sender: zaluzec-at-aaem.amc.anl.gov (Unverified) 9, 15 -- Message-Id: {p06110411bef31161b153-at-[146.139.72.105]} 9, 15 -- Date: Thu, 7 Jul 2005 12:06:10 -0500 9, 15 -- To: microscopy-at-microscopy.com 9, 15 -- From: uti-at-direcpc.com (by way of Nestor J. Zaluzec) 9, 15 -- Subject: [Microscopy] Zeiss EM10C - Roentgen meter board 9, 15 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (vincent.metzger-at-philips.com) from http://www.microscopy.com/MLFormMail.html on Thursday, July 7, 2005 at 11:26:23 ---------------------------------------------------------------------------
Email: vincent.metzger-at-philips.com Name: Vincent Metzger
Organization: philips
Title-Subject: [Filtered] Inca energy - real K-ratio ?
Question: I'm looking for thickness measurement on a stratified sample by using HT variation on SEM and a software to modelize theoricaly the interactions in the stratified sample. I need to extract real Kratio. I made some tests by passing a standart of silicium (for instance), then my sample (containig a strat of Si) and making the ratio between the 2 intensity. This work quite well, but rather time consuming. I standardized my silicium in the inca energy software, but the K-ratio displayed was totally wrong. As an example the system uses Co for quant optimization, and when you analyse the Co, a 1.6 ratio is found:crazy.
I'm wondering how the soft works, very ergonomic but rather a black box. What coefficient is applied? How to get rid of? Is there a option that can be disabled or something else?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (yaseki-at-ucsd.edu) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, July 7, 2005 at 18:25:53 ---------------------------------------------------------------------------
Email: yaseki-at-ucsd.edu Name: Seki Yasuaki
Organization: University of Calfornia-San Diego
Education: Graduate College
Location: La Jolla, CA, USA
Question: Looking for vacuum schemtics, connection help for LEO 438VP SEM. The disassembled instrument has an Edwards Turbo backed by Edwards RV12 pump. The isolation block has an additional port (NW25 fitting) for connection to the specimen chamber. The microscope also has a solenoid operated PV25EK valve. I am looking for info on where this valve is to be located? a) between the block and turbo? or b) between the block and the specimen chamber? or c) elsewher? Any pictures would be of immense help as well.
My apologies if this posting a repeater- I had problems with my e-mail in the past week.
Two similar models of this monitor were available: IBM T221 and Viewsonic VP2290B. Both (I was told) used IBM display panel 22" diagonal, 3840 x 2400 pixels, 204 dpi. Both discontinued. I was also told that IBM had plans of replacing T221 with a new model as early as March 2005. I don't know whether they did yet.
We purchased several such monitors, IBM and Viewsonic, for TEM camera systems. Both models are similar in performance, except Viewsonic is a bit slower in full resolution mode, which makes no difference for static images. It's hard for a PC to run 9.2 MP frames at over 20 FPS refresh rate anyway. In fact, it is better to run 1600 x 1200 desktop most of the time and switch to 3840 x 2400 for high resolution viewing. 1600 x 1200 keeps fonts and icons size comfortable, and refresh rate is fast. The display in 3840 x 2400 mode is stunning- like looking through a window on a sunny day. You can take an eye loupe to the screen and see further detail in the image.
If you will be able to find refurbished or second hand T221 - IBM will honor original 3 year factory warranty as long as monitor is not physically damaged. One of our IBM monitors was refurbished, with a screen defect. IBM replaced the monitor. Consult with IBM regarding the warranty, before buying used monitor. Have monitor serial number when calling IBM. These monitors are still available, mostly on e-bay, and through some internet outlets. Could be even new in box, but not from a regular source.
Is anybody at IBM reading this message? Please comment on a future availability of equal or better display monitor.
See IBM paper on T221 in EM application at http://www-1.ibm.com/industries/healthcare/doc/content/bin/ls_t221_enhancing_electron_1.pdf
Vitaly Feingold SIA 2773 Heath Lane Duluth, GA 30096 Ph. 770-232-7785 Fax 770-232-1791 www.sia-cam.com
----- Original Message ----- X-from: "Mardinly, John" {john.mardinly-at-intel.com} To: "Don Chernoff at ASM" {donc-at-asmicro.com} ; "Microscopy List" {microscopy-at-microscopy.com} Sent: Saturday, July 02, 2005 3:53 AM
Vitally; Lucky you for getting these monitors. At $8400 for the monitor and $2500 for the video card, not too many of us can get the funds for one, but hey, compared to the price a new TEM, why don't new TEMs come standard with at least TWO of these? I have yet to even see one. IBM told me that the T220 was the original 9 megapixel monitor. It was discontinued and then revived as the T221 with a 48 hertz refresh rate and a font handling utility so that fonts could be displayed in a readable size when the monitor was in 3840x2400 mode. What I don't understand, is that over a month after they refused to sell one to me because it was discontinued, it is still listed for sale on the IBM web site! http://www-1.ibm.com/servers/intellistation/pro/t221/index.html
John Mardinly Intel
The opinions of this author do not necessarily represent the opinions on Intel Corporation.
-----Original Message----- X-from: Vitaly Feingold [mailto:vitalylazar-at-att.net] Sent: Thursday, July 07, 2005 11:40 PM To: Microscopy Listserver; Don Chernoff at ASM; Mardinly, John
Hi,
We have troubles with evaporation control unit BSV 202 of our BALZERS BAF 301 FE device. Does anybody have some experiences with it?
The current needed for carbon evaporation suddenly jumped so high that the 8 A fuse blown out happend. After replacing it, the current needed for carbon evaporation stays too high and exceeds the permitted current of safeguarding 8A fuse. There are no differences if we use evaporator 1 (transformer 1) or evaporator 2 (transformer 2). For carbon evaporation we use sharpenned carbon rods.
Thanking in advance for any suggestion. Oldrich ------------------------------------------------------------------------- Oldrich Benada Institute of Microbiology, Acad. Sci. CR Lab. EM Videnska 1083 142 20 Prague 4 Czech Republic
==============================Original Headers============================== 5, 15 -- From benada-at-biomed.cas.cz Fri Jul 8 04:23:11 2005 5, 15 -- Received: from mail.biomed.cas.cz (mail.biomed.cas.cz [147.231.40.5]) 5, 15 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j689NBE8007185 5, 15 -- for {Microscopy-at-microscopy.com} ; Fri, 8 Jul 2005 04:23:11 -0500 5, 15 -- Received: from mail (mail.biomed.cas.cz [147.231.40.5]) 5, 15 -- by mail.biomed.cas.cz (Postfix) with SMTP id 13C9C23D7D 5, 15 -- for {Microscopy-at-microscopy.com} ; Fri, 8 Jul 2005 10:54:27 +0200 (CEST) 5, 15 -- Date: Fri, 8 Jul 2005 10:54:26 +0200 (CEST) 5, 15 -- From: Oldrich Benada {benada-at-biomed.cas.cz} 5, 15 -- X-X-Sender: benada-at-localhost.localdomain 5, 15 -- To: Microscopy-at-microscopy.com 5, 15 -- Subject: Carbon evaporation troubles 5, 15 -- Message-ID: {Pine.LNX.4.44.0507081042510.8516-100000-at-localhost.localdomain} 5, 15 -- MIME-Version: 1.0 5, 15 -- Content-Type: TEXT/PLAIN; charset=US-ASCII ==============================End of - Headers==============================
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O.k., I'm a materials neophyte and I'm looking for some direction. I can ultramicrotome things, I can crush and fracture things, and I can physically polish things, but now I am facing a new sample area: How do I prepare thin films, inorganic, crystal and semi-crystaline grown on a hard smooth substraight (glass, Si, Mica, etc.)? Film thickness below 1-micrometer, mostly below 300nm. We would like to study the morphology of the film via SEM or TEM. EDS and EBSD would also be nice. So how do I get a clean cross-section of the thin film? Buying a dual-column/beam FIB/SEM might be a nice idea but seems a little over-kill, eh? Can someone point me in the right direction? Ion-beam milling? Ion- beam polishing? Sectioning glass or Si crystal just does not sound like a good idea.
Next are there any thoughts on buying equipment and doing this in house vs having samples preped off-campus (I acknowledge that there are some techniques and equipment that are just really tricky).
Thank you!
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Director 350 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
"RAM disk is NOT an installation procedure."
==============================Original Headers============================== 7, 23 -- From edelmare-at-muohio.edu Fri Jul 8 09:52:35 2005 7, 23 -- Received: from mulnx12.mcs.muohio.edu (mulnx12.mcs.muohio.edu [134.53.6.67]) 7, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j68EqYFA030065 7, 23 -- for {microscopy-at-Microscopy.com} ; Fri, 8 Jul 2005 09:52:34 -0500 7, 23 -- Received: from muw2k04 (muw2k04.mcs.muohio.edu [134.53.6.18]) 7, 23 -- by mulnx12.mcs.muohio.edu (Switch-3.1.6/Switch-3.1.6) with SMTP id j68EqWp4015970 7, 23 -- for {microscopy-at-Microscopy.com} ; Fri, 8 Jul 2005 10:52:33 -0400 7, 23 -- Received: From mcsaix06.mcs.muohio.edu ([134.53.253.28]) by muw2k04 (WebShield SMTP v4.5 MR1a P0803.345); 7, 23 -- id 1120834077984; Fri, 8 Jul 2005 10:47:57 -0400 7, 23 -- Received: from emf03 ([134.53.14.119]) 7, 23 -- by mcsaix06.mcs.muohio.edu (Switch-3.1.6/Switch-3.1.6) with ESMTP id j68EqVJu075820 7, 23 -- for {microscopy-at-Microscopy.com} ; Fri, 8 Jul 2005 10:52:31 -0400 7, 23 -- From: "Richard E. Edelmann" {edelmare-at-muohio.edu} 7, 23 -- To: microscopy-at-Microscopy.com 7, 23 -- Date: Fri, 8 Jul 2005 10:52:31 -0400 7, 23 -- MIME-Version: 1.0 7, 23 -- Content-type: text/plain; charset=US-ASCII 7, 23 -- Content-transfer-encoding: 7BIT 7, 23 -- Subject: Imaging thin films 7, 23 -- Message-ID: {42CE5AEF.1499.3CAEB37-at-localhost} 7, 23 -- Priority: normal 7, 23 -- X-mailer: Pegasus Mail for Win32 (v3.12c) 7, 23 -- X-Scanned-By: MIMEDefang 2.45 ==============================End of - Headers==============================
I have been following this thread with interest, as it affects us of course. We would like to use higher resolution monitors for our TEM cameras, but perhaps you could answer two questions for me:
1) What exactly would be the advantage of a high resolution monitor? Unless you can see the pixilation on a regular monitor, a higher resolution monitor would not show much more. To see more, the screen would also have to be much bigger. Of course, you take a magnifying glass to the monitor and see more, but that you can do with a zoom function on a regular monitor as well.
2) Is this worth around $10K or more to you?
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----- X-from: john.mardinly-at-intel.com [mailto:john.mardinly-at-intel.com] Sent: Friday, July 08, 2005 1:29 AM To: Mike Bode
Vitally; Lucky you for getting these monitors. At $8400 for the monitor and $2500 for the video card, not too many of us can get the funds for one, but hey, compared to the price a new TEM, why don't new TEMs come standard with at least TWO of these? I have yet to even see one. IBM told me that the T220 was the original 9 megapixel monitor. It was discontinued and then revived as the T221 with a 48 hertz refresh rate and a font handling utility so that fonts could be displayed in a readable size when the monitor was in 3840x2400 mode. What I don't understand, is that over a month after they refused to sell one to me because it was discontinued, it is still listed for sale on the IBM web site! http://www-1.ibm.com/servers/intellistation/pro/t221/index.html
John Mardinly Intel
The opinions of this author do not necessarily represent the opinions on Intel Corporation.
-----Original Message----- X-from: Vitaly Feingold [mailto:vitalylazar-at-att.net] Sent: Thursday, July 07, 2005 11:40 PM To: Microscopy Listserver; Don Chernoff at ASM; Mardinly, John
Hi Nestor,
As of today I am getting all postings twice. Is that a problem with my subscription or a general problem?
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 Headers============================== 6, 21 -- From Mike.Bode-at-soft-imaging.net Fri Jul 8 10:08:39 2005 6, 21 -- Received: from dr-lnx1.soft-imaging.com (67.104.115.34.ptr.us.xo.net [67.104.115.34]) 6, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j68F8dRr013305 6, 21 -- for {Microscopy-at-microscopy.com} ; Fri, 8 Jul 2005 10:08:39 -0500 6, 21 -- Received: from lakewood.soft-imaging.com (lakewood.soft-imaging.com [192.168.5.225]) 6, 21 -- by dr-lnx1.soft-imaging.com (8.11.6/8.11.6/SuSE Linux 0.5) with ESMTP id j68F8bx00071 6, 21 -- for {Microscopy-at-microscopy.com} ; Fri, 8 Jul 2005 09:08:37 -0600 6, 21 -- Received: by hq-dc2.soft-imaging.net with Internet Mail Service (5.5.2657.72) 6, 21 -- id {3GTHZ2J5} ; Fri, 8 Jul 2005 09:08:55 -0600 6, 21 -- Message-ID: {6127CE87B9BDD511B59D0001028A497D011C226A-at-hq-dc2.soft-imaging.net} 6, 21 -- From: Mike Bode {Mike.Bode-at-soft-imaging.net} 6, 21 -- To: "'Microscopy-at-microscopy.com'" {Microscopy-at-microscopy.com} 6, 21 -- Subject: For Nestor: Double posting 6, 21 -- Date: Fri, 8 Jul 2005 09:07:53 -0600 6, 21 -- Deferred-Delivery: Fri, 8 Jul 2005 09:07:00 -0600 6, 21 -- MIME-Version: 1.0 6, 21 -- X-Mailer: Internet Mail Service (5.5.2657.72) 6, 21 -- Content-Type: text/plain; 6, 21 -- charset="iso-8859-1" 6, 21 -- Content-Transfer-Encoding: 8bit 6, 21 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id j68F8dRr013305 ==============================End of - Headers==============================
SEM will produce morphology info whereas EBSD will produce grain, orientation and texture info. Totally different results and prep methods.
Depending on what the film material is, that would dictate how it is prepared for analysis. Silicon is easiest I think to use as a substrate since it can easily be cut into small pieces. Mount the piece and coat it then SEM image for morphology. Mount another piece and polish it with colloidal silica or alumina (again, depending on film material) down to .02u and this should work for EBSD.
FIB ablades the surface and does not produce good EBSD specimens. I've tried various plasmas and Gatan 682 and do not get good specimens for EBSD. There is probably some magic combination that I have yet to find. Thus far, mechanical polishing and slight etch with DI water seems to do the job. There are dedicated ion beam polishing tools that do nice jobs for EBSD. But unless you are going to do a lot of this, I don't think it pencils out.
Since the analysis is tops down, I don't see why you would need to section the specimen. If there is more to your situation, tell us more.
gary g.
At 07:55 AM 7/8/2005, you wrote:
} O.k., I'm a materials neophyte and I'm looking for some } direction. I can ultramicrotome things, I can crush and fracture } things, and I can physically polish things, but now I am facing a new } sample area: How do I prepare thin films, inorganic, crystal and } semi-crystaline grown on a hard smooth substraight (glass, Si, } Mica, etc.)? Film thickness below 1-micrometer, mostly below } 300nm. We would like to study the morphology of the film via SEM } or TEM. EDS and EBSD would also be nice. So how do I get a } clean cross-section of the thin film? Buying a dual-column/beam } FIB/SEM might be a nice idea but seems a little over-kill, eh? Can } someone point me in the right direction? Ion-beam milling? Ion- } beam polishing? Sectioning glass or Si crystal just does not sound } like a good idea. } } Next are there any thoughts on buying equipment and doing this in } house vs having samples preped off-campus (I acknowledge that } there are some techniques and equipment that are just really tricky). } } Thank you! } } } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Director } 350 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } http://www.emf.muohio.edu
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (biology-at-ucla.edu) from http://www.microscopy.org/MicroscopyListserver/MLFormMail.html on Friday, July 8, 2005 at 10:41:07 ---------------------------------------------------------------------------
Email: biology-at-ucla.edu Name: Eric A. Rosen
Organization: UCLA Medical Center
Title-Subject: I am trying to get a hold of Judy Murphy at the San Joaquin Delta
Question: I am trying to get a hold of Judy Murphy at the San Joaquin Delta College EM School I have a open EM position and would like to advertise it with her...
I am not quite sure what it means when you say that the resolution of the human eye is 300 DPI. I looked up an article on the web (http://www.madsci.org/posts/archives/may97/864446241.Ph.r.html), which specifies the angular resolution of the human eye under optimum conditions as 1/60th of a degree. If I do my math right, the resolution is then given by:
2 x distance x tan (angle/2).
For a viewing distance of about 1m, I get a resolution of 0.3mm (which corresponds to about 220 DPI at 1m distance), which is more or less the dot pitch of a normal monitor (a 21" LCD at 1600x1200 has a dot pitch of 0.294mm). So, I think the normal monitors are actually at the limits of the human eye's resolution. If your eyes were much better, you should be able to see the individual red, green, and blue dots from a distance of 1m.
As I said, the optimum resolution assumes certain parameters: 25 cm viewing distance, perfect lighting, etc. All changes will reduce the resolution.
But that's all mathematics. Has anybody actually compared a high res monitor and a regular monitor side by side?
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----- X-from: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Friday, July 08, 2005 10:04 AM To: Mike Bode
Hi all,
I have been following this thread with interest, as it affects us of course. We would like to use higher resolution monitors for our TEM cameras, but perhaps you could answer two questions for me:
1) What exactly would be the advantage of a high resolution monitor? Unless you can see the pixilation on a regular monitor, a higher resolution monitor would not show much more. To see more, the screen would also have to be much bigger. Of course, you take a magnifying glass to the monitor and see more, but that you can do with a zoom function on a regular monitor as well.
2) Is this worth around $10K or more to you?
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----- X-from: john.mardinly-at-intel.com [mailto:john.mardinly-at-intel.com] Sent: Friday, July 08, 2005 1:29 AM To: Mike Bode
Vitally; Lucky you for getting these monitors. At $8400 for the monitor and $2500 for the video card, not too many of us can get the funds for one, but hey, compared to the price a new TEM, why don't new TEMs come standard with at least TWO of these? I have yet to even see one. IBM told me that the T220 was the original 9 megapixel monitor. It was discontinued and then revived as the T221 with a 48 hertz refresh rate and a font handling utility so that fonts could be displayed in a readable size when the monitor was in 3840x2400 mode. What I don't understand, is that over a month after they refused to sell one to me because it was discontinued, it is still listed for sale on the IBM web site! http://www-1.ibm.com/servers/intellistation/pro/t221/index.html
John Mardinly Intel
The opinions of this author do not necessarily represent the opinions on Intel Corporation.
-----Original Message----- X-from: Vitaly Feingold [mailto:vitalylazar-at-att.net] Sent: Thursday, July 07, 2005 11:40 PM To: Microscopy Listserver; Don Chernoff at ASM; Mardinly, John
I am considering buying a paraffin microtome for our core facility. I don't personally use one much so if any knowledgeable users have recommendations on brands or models to buy (or avoid), i would welcome replies (probably best sent to me directly and not the listserver). I will be happy to keep your comments confidential. In addition, if there are special features you think are essential, i would be pleased to hear of them also. thanks, tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
Mike; One meter viewing distance? Sorry, my arms are no where near that long. Fortunately my presbyopia is compensated by severe myopia. Otherwise, use reading glasses. Closer up (~1 foot?), the human eye can resolve ~80-100 microns, which is 250-300 DPI, which is why probably why 300 DPI has been a target spec for printers for a long time. The IBM T221 has a pixel pitch of 0.1245 millimeters, so those pixels are half the size of the pixels of garden variety monitors that cost less, and the way that translates into 200DPI is (25 mm/in)/(0.1245 mm/pixel)=200.8 pixels/inch. BTW, I can see the vertical mask stripes of my Sony monitor with my naked eye if I get close, and everybody in our lab could see the difference between a 400DPI Fuji Pictrograph print and a monitor display.
John Mardinly Intel
The opinions of this author do not necessarily represent the opinions on Intel Corporation.
-----Original Message----- X-from: Mike.Bode-at-soft-imaging.net [mailto:Mike.Bode-at-soft-imaging.net] Sent: Friday, July 08, 2005 9:46 AM To: Mardinly, John
Hello John,
I am not quite sure what it means when you say that the resolution of the human eye is 300 DPI. I looked up an article on the web (http://www.madsci.org/posts/archives/may97/864446241.Ph.r.html), which specifies the angular resolution of the human eye under optimum conditions as 1/60th of a degree. If I do my math right, the resolution is then given by:
2 x distance x tan (angle/2).
For a viewing distance of about 1m, I get a resolution of 0.3mm (which corresponds to about 220 DPI at 1m distance), which is more or less the dot pitch of a normal monitor (a 21" LCD at 1600x1200 has a dot pitch of 0.294mm). So, I think the normal monitors are actually at the limits of the human eye's resolution. If your eyes were much better, you should be able to see the individual red, green, and blue dots from a distance of 1m.
As I said, the optimum resolution assumes certain parameters: 25 cm viewing distance, perfect lighting, etc. All changes will reduce the resolution.
But that's all mathematics. Has anybody actually compared a high res monitor and a regular monitor side by side?
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----- X-from: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Friday, July 08, 2005 10:04 AM To: Mike Bode
Hi all,
I have been following this thread with interest, as it affects us of course. We would like to use higher resolution monitors for our TEM cameras, but perhaps you could answer two questions for me:
1) What exactly would be the advantage of a high resolution monitor? Unless you can see the pixilation on a regular monitor, a higher resolution monitor would not show much more. To see more, the screen would also have to be much bigger. Of course, you take a magnifying glass to the monitor and see more, but that you can do with a zoom function on a regular monitor as well.
2) Is this worth around $10K or more to you?
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----- X-from: john.mardinly-at-intel.com [mailto:john.mardinly-at-intel.com] Sent: Friday, July 08, 2005 1:29 AM To: Mike Bode
Vitally; Lucky you for getting these monitors. At $8400 for the monitor and $2500 for the video card, not too many of us can get the funds for one, but hey, compared to the price a new TEM, why don't new TEMs come standard with at least TWO of these? I have yet to even see one. IBM told me that the T220 was the original 9 megapixel monitor. It was discontinued and then revived as the T221 with a 48 hertz refresh rate and a font handling utility so that fonts could be displayed in a readable size when the monitor was in 3840x2400 mode. What I don't understand, is that over a month after they refused to sell one to me because it was discontinued, it is still listed for sale on the IBM web site! http://www-1.ibm.com/servers/intellistation/pro/t221/index.html
John Mardinly Intel
The opinions of this author do not necessarily represent the opinions on Intel Corporation.
-----Original Message----- X-from: Vitaly Feingold [mailto:vitalylazar-at-att.net] Sent: Thursday, July 07, 2005 11:40 PM To: Microscopy Listserver; Don Chernoff at ASM; Mardinly, John
Hi John,
I was actually talking about monitors. I agree with you that you normally look at prints at a shorter distance, but I think 1 m for a monitor is a reasonable distance.
And what you are saying ("BTW, I can see the vertical mask stripes of my Sony monitor with my naked eye if I get close") basically confirms my assumption: If you are NOT up close, you can't see the stripe, so the pixel size is close to resolution of the eye (at normal viewing distances). You would not be able to see or detect a further reduction in pixel size. I don't know if the width of the vertical mask is the same as a pixel. I, for one, cannot see individual green, red and blue pixels when I look at a white spot on the monitor, unless I use a magnifying glass.
Comparing printers and monitors is difficult. One is "back-illuminated", the other shows reflected light, the color gamut is different, etc. I'd be interested to hear from someone who has compared a normal LCD monitor with a high resolution LCD monitor.
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----- X-from: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Friday, July 08, 2005 1:11 PM To: Mike Bode Cc: Listserver
Hello John,
I am not quite sure what it means when you say that the resolution of the human eye is 300 DPI. I looked up an article on the web (http://www.madsci.org/posts/archives/may97/864446241.Ph.r.html), which specifies the angular resolution of the human eye under optimum conditions as 1/60th of a degree. If I do my math right, the resolution is then given by:
2 x distance x tan (angle/2).
For a viewing distance of about 1m, I get a resolution of 0.3mm (which corresponds to about 220 DPI at 1m distance), which is more or less the dot pitch of a normal monitor (a 21" LCD at 1600x1200 has a dot pitch of 0.294mm). So, I think the normal monitors are actually at the limits of the human eye's resolution. If your eyes were much better, you should be able to see the individual red, green, and blue dots from a distance of 1m.
As I said, the optimum resolution assumes certain parameters: 25 cm viewing distance, perfect lighting, etc. All changes will reduce the resolution.
But that's all mathematics. Has anybody actually compared a high res monitor and a regular monitor side by side?
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----- X-from: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Friday, July 08, 2005 10:04 AM To: Mike Bode
Hi all,
I have been following this thread with interest, as it affects us of course. We would like to use higher resolution monitors for our TEM cameras, but perhaps you could answer two questions for me:
1) What exactly would be the advantage of a high resolution monitor? Unless you can see the pixilation on a regular monitor, a higher resolution monitor would not show much more. To see more, the screen would also have to be much bigger. Of course, you take a magnifying glass to the monitor and see more, but that you can do with a zoom function on a regular monitor as well.
2) Is this worth around $10K or more to you?
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----- X-from: john.mardinly-at-intel.com [mailto:john.mardinly-at-intel.com] Sent: Friday, July 08, 2005 1:29 AM To: Mike Bode
Vitally; Lucky you for getting these monitors. At $8400 for the monitor and $2500 for the video card, not too many of us can get the funds for one, but hey, compared to the price a new TEM, why don't new TEMs come standard with at least TWO of these? I have yet to even see one. IBM told me that the T220 was the original 9 megapixel monitor. It was discontinued and then revived as the T221 with a 48 hertz refresh rate and a font handling utility so that fonts could be displayed in a readable size when the monitor was in 3840x2400 mode. What I don't understand, is that over a month after they refused to sell one to me because it was discontinued, it is still listed for sale on the IBM web site! http://www-1.ibm.com/servers/intellistation/pro/t221/index.html
John Mardinly Intel
The opinions of this author do not necessarily represent the opinions on Intel Corporation.
-----Original Message----- X-from: Vitaly Feingold [mailto:vitalylazar-at-att.net] Sent: Thursday, July 07, 2005 11:40 PM To: Microscopy Listserver; Don Chernoff at ASM; Mardinly, John
Mike; Phil Batson reported at M&M 2004 that it was extremely useful and highly recommended. Vitaly Feingold reported this week in the Listserver that he has two. Perhaps he can also describe the experience of beholding an IBM T221.
John Mardinly Intel
The opinions of this author do not necessarily represent the opinions on Intel Corporation.
-----Original Message----- X-from: Mike.Bode-at-soft-imaging.net [mailto:Mike.Bode-at-soft-imaging.net] Sent: Friday, July 08, 2005 12:49 PM To: Mardinly, John
Hi John,
I was actually talking about monitors. I agree with you that you normally look at prints at a shorter distance, but I think 1 m for a monitor is a reasonable distance.
And what you are saying ("BTW, I can see the vertical mask stripes of my Sony monitor with my naked eye if I get close") basically confirms my assumption: If you are NOT up close, you can't see the stripe, so the pixel size is close to resolution of the eye (at normal viewing distances). You would not be able to see or detect a further reduction in pixel size. I don't know if the width of the vertical mask is the same as a pixel. I, for one, cannot see individual green, red and blue pixels when I look at a white spot on the monitor, unless I use a magnifying glass.
Comparing printers and monitors is difficult. One is "back-illuminated", the other shows reflected light, the color gamut is different, etc. I'd be interested to hear from someone who has compared a normal LCD monitor with a high resolution LCD monitor.
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----- X-from: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Friday, July 08, 2005 1:11 PM To: Mike Bode Cc: Listserver
Hello John,
I am not quite sure what it means when you say that the resolution of the human eye is 300 DPI. I looked up an article on the web (http://www.madsci.org/posts/archives/may97/864446241.Ph.r.html), which specifies the angular resolution of the human eye under optimum conditions as 1/60th of a degree. If I do my math right, the resolution is then given by:
2 x distance x tan (angle/2).
For a viewing distance of about 1m, I get a resolution of 0.3mm (which corresponds to about 220 DPI at 1m distance), which is more or less the dot pitch of a normal monitor (a 21" LCD at 1600x1200 has a dot pitch of 0.294mm). So, I think the normal monitors are actually at the limits of the human eye's resolution. If your eyes were much better, you should be able to see the individual red, green, and blue dots from a distance of 1m.
As I said, the optimum resolution assumes certain parameters: 25 cm viewing distance, perfect lighting, etc. All changes will reduce the resolution.
But that's all mathematics. Has anybody actually compared a high res monitor and a regular monitor side by side?
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----- X-from: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Friday, July 08, 2005 10:04 AM To: Mike Bode
Hi all,
I have been following this thread with interest, as it affects us of course. We would like to use higher resolution monitors for our TEM cameras, but perhaps you could answer two questions for me:
1) What exactly would be the advantage of a high resolution monitor? Unless you can see the pixilation on a regular monitor, a higher resolution monitor would not show much more. To see more, the screen would also have to be much bigger. Of course, you take a magnifying glass to the monitor and see more, but that you can do with a zoom function on a regular monitor as well.
2) Is this worth around $10K or more to you?
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----- X-from: john.mardinly-at-intel.com [mailto:john.mardinly-at-intel.com] Sent: Friday, July 08, 2005 1:29 AM To: Mike Bode
Vitally; Lucky you for getting these monitors. At $8400 for the monitor and $2500 for the video card, not too many of us can get the funds for one, but hey, compared to the price a new TEM, why don't new TEMs come standard with at least TWO of these? I have yet to even see one. IBM told me that the T220 was the original 9 megapixel monitor. It was discontinued and then revived as the T221 with a 48 hertz refresh rate and a font handling utility so that fonts could be displayed in a readable size when the monitor was in 3840x2400 mode. What I don't understand, is that over a month after they refused to sell one to me because it was discontinued, it is still listed for sale on the IBM web site! http://www-1.ibm.com/servers/intellistation/pro/t221/index.html
John Mardinly Intel
The opinions of this author do not necessarily represent the opinions on Intel Corporation.
-----Original Message----- X-from: Vitaly Feingold [mailto:vitalylazar-at-att.net] Sent: Thursday, July 07, 2005 11:40 PM To: Microscopy Listserver; Don Chernoff at ASM; Mardinly, John
John et al.:
A few more details on what I wrote on this subject before. Lyson has been making inks and papers for photo quality ink jet printing for some time. Their new system is the "Daylight Darkroom". It is a dedicated 4 to 7 ink system (depending on your printer) for black and white printing only. I think the price includes software and inks but no printer, the system only works with certain printers. It has gotten very favorable reviews in fine art photo magazines (May/June 2005 issue of Photo Techniques, the review might be on the magazine's website (www.phototechmag.com) or on Lyson's). Lyson will send you some sample prints if you register at their website. I suspect that if someone sent Lyson a digital TEM or SEM image they might make a print of that as a sales tool. I'll bet they have no idea that there is a market for their products in the EM field. I only wish I did more than a few prints per year of parts of ground up cells. Lyson is not the only firm making inks, papers and printing systems for fine art B&W photography. Check out InkjetMall.com or MediaStreet.com. Disclaimer: I don't have any financial interest in any of the firms or products I mentioned.
Geoff
John J. Bozzola wrote:
} ------------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------------- } } } We are comparing dye sub printers versus the Fuji Pictrography PG4500. } The price differential is a staggering $22,000 versus $6,500, } respectively. It is my understanding that both produce good quality, } gray-scale prints (versus dithered, inkjet prints). Why the price } differential? I would welcome any comments, user experiences, etc. } Does anyone know the average cost per 8.5 x 11in print? } } Thank you.
--
-- ********************************************** 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 **********************************************
==============================Original Headers============================== 10, 32 -- From mcauliff-at-umdnj.edu Fri Jul 8 16:04:45 2005 10, 32 -- Received: from mail01.umdnj.edu (zix01.UMDNJ.EDU [130.219.34.124]) 10, 32 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j68L4jG7014722 10, 32 -- for {Microscopy-at-msa.microscopy.com} ; Fri, 8 Jul 2005 16:04:45 -0500 10, 32 -- Received: from zix01.umdnj.edu (ZixVPM [127.0.0.1]) 10, 32 -- by Outbound.umdnj.edu (Proprietary) with ESMTP id 13C3FEC05E 10, 32 -- for {Microscopy-at-msa.microscopy.com} ; Fri, 8 Jul 2005 17:04:45 -0400 (EDT) 10, 32 -- Received: from polaris.umdnj.edu (polarisa.UMDNJ.EDU [130.219.34.131]) 10, 32 -- by mail01.umdnj.edu (Proprietary) with ESMTP id 0239DEC057 10, 32 -- for {Microscopy-at-msa.microscopy.com} ; Fri, 8 Jul 2005 17:04:44 -0400 (EDT) 10, 32 -- Received: from conversion-daemon.Polaris.umdnj.edu by Polaris.umdnj.edu 10, 32 -- (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 2004)) 10, 32 -- id {0IJB00E01TI1QW-at-Polaris.umdnj.edu} (original mail from mcauliff-at-umdnj.edu) 10, 32 -- for Microscopy-at-msa.microscopy.com; Fri, 08 Jul 2005 17:04:43 -0400 (EDT) 10, 32 -- Received: from [127.0.0.1] ([10.138.2.240]) 10, 32 -- by Polaris.umdnj.edu (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 10, 32 -- 2004)) with ESMTP id {0IJB002G2UJV7T-at-Polaris.umdnj.edu} ; Fri, 10, 32 -- 08 Jul 2005 17:04:43 -0400 (EDT) 10, 32 -- Date: Fri, 08 Jul 2005 17:04:52 -0400 10, 32 -- From: Geoff McAuliffe {mcauliff-at-umdnj.edu} 10, 32 -- Subject: Re: [Microscopy] ink jet versus dye sub versus Pictrography 10, 32 -- In-reply-to: {p06110419beea0f7e2027-at-[131.230.177.142]} 10, 32 -- To: "John J. Bozzola" {bozzola-at-siu.edu} 10, 32 -- Cc: Microscopy-at-msa.microscopy.com 10, 32 -- Message-id: {42CEEA74.40601-at-umdnj.edu} 10, 32 -- MIME-version: 1.0 10, 32 -- Content-type: text/plain; format=flowed; charset=ISO-8859-1 10, 32 -- Content-transfer-encoding: 7BIT 10, 32 -- X-Accept-Language: en-us, en 10, 32 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 10, 32 -- Gecko/20040804 Netscape/7.2 (ax) 10, 32 -- References: {p06110419beea0f7e2027-at-[131.230.177.142]} ==============================End of - Headers==============================
Richard E. Edelmann wrote: =========================================== O.k., I'm a materials neophyte and I'm looking for some direction. I can ultramicrotome things, I can crush and fracture things, and I can physically polish things, but now I am facing a new sample area: How do I prepare thin films, inorganic, crystal and semi-crystaline grown on a hard smooth substraight (glass, Si, Mica, etc.)? Film thickness below 1-micrometer, mostly below 300nm. We would like to study the morphology of the film via SEM or TEM. EDS and EBSD would also be nice. So how do I get a clean cross-section of the thin film? Buying a dual-column/beam FIB/SEM might be a nice idea but seems a little over-kill, eh? Can someone point me in the right direction? Ion-beam milling? Ion- beam polishing? Sectioning glass or Si crystal just does not sound like a good idea.
Next are there any thoughts on buying equipment and doing this in house vs having samples preped off-campus (I acknowledge that there are some techniques and equipment that are just really tricky). =============================================== Since you are already set up to do ultramicrotomy, have you considered the following:
a) Take a freshly cleaved stripping of HOPG and then do your evaporation. If you are not familiar with HOPG, see URL http://www.2spi.com/catalog/new/hopgsub.shtml In terms of smoothness, HOPG can be atomically smooth, the size of the atomically smooth areas depending on the grade of the HOPG selected.
b) Put the HOPG stripping in a flat embedding mold and then expose the coated sample, HOPG side up, to an oxygen plasma, such as in one of our Plasma Prep II plasma etchers, which will etch away the HOPG but leave your coating intact.
c) Fill the cavity containing the thin film, now with HOPG removed, with your favorite Epon 812 substitute, or SPI-Pon 812 and after curing,
d) Face off the block and diamond knife thin section the coating.
A variation on this theme would be to substitute a freshly cleaved surface of NaCl for the substrate, and after coating, the NaCl is dissolved away and the coating when dry, can be embedded as above.
You should have no difficulty getting good TEM images of the coating.
We actually prefer diamond knife thin sectioning to other methods for this kind of sample. It is not a method free of artifacts. But when they are ones that are induced by the knife edge itself, they are directional and easily recognized as being artifacts, but for the other approaches one might consider, the artifacts are not directional and therefore much more difficult to recognize.
Disclaimer: SPI Supplies offers HOPG, plasma etchers, diamond knives, and NaCl crystals so we would have a vested interest in promoting this approach relative to others.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 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 ############################ ==================================================
==============================Original Headers============================== 15, 21 -- From cgarber-at-2spi.com Fri Jul 8 17:02:14 2005 15, 21 -- Received: from diskless1.axs2000.net (mail.del.net [209.120.196.47]) 15, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j68M2Ek0023153 15, 21 -- for {microscopy-at-msa.microscopy.com} ; Fri, 8 Jul 2005 17:02:14 -0500 15, 21 -- Received: from gw.idv.net (diskless6.external [209.120.196.50]) 15, 21 -- by diskless1.axs2000.net (8.12.11/8.12.11) with ESMTP id j68M2Dmp009029; 15, 21 -- Fri, 8 Jul 2005 18:02:13 -0400 15, 21 -- X-IDV-FirstRcvd: diskless6.external [209.120.196.50] 15, 21 -- X-IDV-HELO: gw.idv.net 15, 21 -- X-Originating-IP: [81.254.170.250] 15, 21 -- From: cgarber-at-2spi.com 15, 21 -- To: microscopy-at-msa.microscopy.com 15, 21 -- Subject: Characterization of thin film coatings by TEM 15, 21 -- Date: Fri, 08 Jul 2005 22:02:14 +0000 15, 21 -- Message-ID: {20050708.UJ5.17172200-at-gw.idv.net} 15, 21 -- MIME-Version: 1.0 15, 21 -- Content-Type: text/plain; 15, 21 -- charset="iso-8859-1" 15, 21 -- Content-Transfer-Encoding: 8bit 15, 21 -- Content-Disposition: inline 15, 21 -- X-Mailer: phpGroupWare (http://www.phpgroupware.org) v 0.9.13.018 ==============================End of - Headers==============================
Oldrich Benada wrote: ================================================================= We have troubles with evaporation control unit BSV 202 of our BALZERS BAF 301 FE device. Does anybody have some experiences with it?
The current needed for carbon evaporation suddenly jumped so high that the 8 A fuse blown out happend. After replacing it, the current needed for carbon evaporation stays too high and exceeds the permitted current of safeguarding 8A fuse. There are no differences if we use evaporator 1 (transformer 1) or evaporator 2 (transformer 2). For carbon evaporation we use sharpenned carbon rods.
Thanking in advance for any suggestion. ================================================================= Have you switched sources for your "carbon" rods lately? First, some vendors offering "carbon" rods are really offering graphite rods. It is my own perception that most people who think they are using carbon rods are actually using graphite rods. Also, even among graphite rods, there is a large variation of resistivities (due to variations in density). So a common problem is that one procures the wrong kind of rods (which would have a resistivity that is out of range for their instrumental set up) so that in order to get evaporation, they exceed the instrumental parameters of their system. See URL http://2spi.com/catalog/spec_prep/carbon-graphite-rods.html for our simple "test" to judge which type of rod you are actually using.
If you have not switched rod vendors, then the second most common reason is that you are not making a sharp enough "point". Hence the tip, in order to get hot enough to evaporate carbon, draws so much current, you exceed the limits of your system. For the optimum dimensions for rod tips, see the drawings on URL http://2spi.com/catalog/spec_prep/carbon-rods.shtml
Rods can be purchased pre-sharpened from all of the major suppliers of consumables for EM laboratories, such as SPI, but if you want to make your own presharpened points, carbon rod sharpeners can be purchased as well such as are found on URL http://2spi.com/catalog/spec_prep/carbon-rod-sharpener.shtml
Disclaimer: SPI Supplies offers carbon and graphite rods as well as sharpeners.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 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 ############################ ==================================================
==============================Original Headers============================== 12, 22 -- From cgarber-at-2spi.com Sat Jul 9 03:09:02 2005 12, 22 -- Received: from smtp-wifi.orange.fr (smtp-wifi.orange.fr [194.250.131.236]) 12, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j69892Um019929 12, 22 -- for {microscopy-at-msa.microscopy.com} ; Sat, 9 Jul 2005 03:09:02 -0500 12, 22 -- Received: from ibm1x23g2abfyg (81.254.170.250) by smtp-wifi.orange.fr (7.0.031.3) 12, 22 -- id 425E7F1100039606 for microscopy-at-msa.microscopy.com; Sat, 9 Jul 2005 10:14:36 +0200 12, 22 -- Message-ID: {00d101c5845d$74ee33c0$faaafe51-at-ibm1x23g2abfyg} 12, 22 -- Reply-To: "Charles A Garber" {cgarber-at-2spi.com} 12, 22 -- From: "Charles A Garber" {cgarber-at-2spi.com} 12, 22 -- To: "Microscopy Listserver" {microscopy-at-msa.microscopy.com} 12, 22 -- Subject: carbon evaporation problem 12, 22 -- Date: Sat, 9 Jul 2005 04:08:57 -0400 12, 22 -- MIME-Version: 1.0 12, 22 -- Content-Type: text/plain; 12, 22 -- format=flowed; 12, 22 -- charset="Windows-1252"; 12, 22 -- reply-type=original 12, 22 -- Content-Transfer-Encoding: 7bit 12, 22 -- X-Priority: 3 12, 22 -- X-MSMail-Priority: Normal 12, 22 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2180 12, 22 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 ==============================End of - Headers==============================
It is unquestionably right that the distance of best resolution for the human eye is about 25 cm. However, I was talking about working on a monitor, and I am typically sitting at arm's length from it. I just measured my arm, and it is 75 cm. I do not work with a monitor with my eyes 25 cm from the screen. Would definitely give me headaches.
I am not trying to prove to everyone that high res monitors are useless, far from it. But I am personally not convinced that it would be worth $10,000. Considering the eye's resolution and the options of today's software, I think you can easily get what you want on a lower res monitor.
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----- X-from: john.mardinly-at-intel.com [mailto:john.mardinly-at-intel.com] Sent: Friday, July 08, 2005 18:50 To: Mike Bode
Barbara; YES! 10 inches is the right number! That's just slightly less than the length of my arms, and slightly more than the length of my nose, so everything I look at ends up at about 10 inches.
John Mardinly Intel
The opinions of this author do not necessarily represent the opinions on Intel Corporation.
-----Original Message----- X-from: Barbara Foster [mailto:bfoster-at-mme1.com] Sent: Friday, July 08, 2005 2:59 PM To: Mardinly, John
==============================Original Headers============================== 19, 38 -- From john.mardinly-at-intel.com Fri Jul 8 19:44:38 2005 19, 38 -- Received: from fmsfmr002.fm.intel.com (fmr14.intel.com [192.55.52.68]) 19, 38 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j690ibow032523 19, 38 -- for {Microscopy-at-microscopy.com} ; Fri, 8 Jul 2005 19:44:38 -0500 19, 38 -- Received: from fmsfmr100.fm.intel.com (fmsfmr100.fm.intel.com [10.253.24.20]) 19, 38 -- by fmsfmr002.fm.intel.com (8.12.10/8.12.10/d: major-outer.mc,v 1.1 2004/09/17 17:50:56 root Exp $) with ESMTP id j690ibos007054; 19, 38 -- Sat, 9 Jul 2005 00:44:37 GMT 19, 38 -- Received: from fmsmsxvs043.fm.intel.com (fmsmsxvs043.fm.intel.com [132.233.42.129]) 19, 38 -- by fmsfmr100.fm.intel.com (8.12.10/8.12.10/d: major-inner.mc,v 1.2 2004/09/17 18:05:01 root Exp $) with SMTP id j690iWaC000506; 19, 38 -- Sat, 9 Jul 2005 00:44:35 GMT 19, 38 -- Received: from fmsmsx332.amr.corp.intel.com ([132.233.42.148]) 19, 38 -- by fmsmsxvs043.fm.intel.com (SAVSMTP 3.1.7.47) with SMTP id M2005070817443407650 19, 38 -- ; Fri, 08 Jul 2005 17:44:34 -0700 19, 38 -- Received: from fmsmsx311.amr.corp.intel.com ([132.233.42.214]) by fmsmsx332.amr.corp.intel.com with Microsoft SMTPSVC(6.0.3790.211); 19, 38 -- Fri, 8 Jul 2005 17:44:35 -0700 19, 38 -- Received: from scsmsx402.amr.corp.intel.com ([10.3.90.16]) by fmsmsx311.amr.corp.intel.com with Microsoft SMTPSVC(6.0.3790.211); 19, 38 -- Fri, 8 Jul 2005 17:44:34 -0700 19, 38 -- Received: from scsmsx403.amr.corp.intel.com ([10.3.90.18]) by scsmsx402.amr.corp.intel.com with Microsoft SMTPSVC(6.0.3790.211); 19, 38 -- Fri, 8 Jul 2005 17:44:33 -0700 19, 38 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 19, 38 -- Content-class: urn:content-classes:message 19, 38 -- MIME-Version: 1.0 19, 38 -- Content-Type: text/plain; 19, 38 -- charset="us-ascii" 19, 38 -- Subject: RE: [Microscopy] RE: RE: RE: RE: RE: Image review: Hard copy vs. PC 19, 38 -- Date: Fri, 8 Jul 2005 17:44:32 -0700 19, 38 -- Message-ID: {9E1ED6A623D8CB44B4866ACF20ECDB2B06D40C2F-at-scsmsx403.amr.corp.intel.com} 19, 38 -- X-MS-Has-Attach: 19, 38 -- X-MS-TNEF-Correlator: 19, 38 -- Thread-Topic: [Microscopy] RE: RE: RE: RE: RE: Image review: Hard copy vs. PC 19, 38 -- Thread-Index: AcWECE7TyeyW6fSsQiW5AVr84prlVQAFmI6A 19, 38 -- From: "Mardinly, John" {john.mardinly-at-intel.com} 19, 38 -- To: "Barbara Foster" {bfoster-at-mme1.com} 19, 38 -- Cc: "Listserver" {Microscopy-at-microscopy.com} 19, 38 -- X-OriginalArrivalTime: 09 Jul 2005 00:44:33.0970 (UTC) FILETIME=[6004B120:01C5841F] 19, 38 -- X-Scanned-By: MIMEDefang 2.44 19, 38 -- Content-Transfer-Encoding: 8bit 19, 38 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id j690ibow032523 ==============================End of - Headers==============================
==============================Original Headers============================== 32, 19 -- From Mike.Bode-at-soft-imaging.net Sat Jul 9 11:48:17 2005 32, 19 -- Received: from dr-lnx1.soft-imaging.com (67.104.115.34.ptr.us.xo.net [67.104.115.34]) 32, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j69GmH8f008366 32, 19 -- for {Microscopy-at-microscopy.com} ; Sat, 9 Jul 2005 11:48:17 -0500 32, 19 -- Received: from lakewood.soft-imaging.com (lakewood.soft-imaging.com [192.168.5.225]) 32, 19 -- by dr-lnx1.soft-imaging.com (8.11.6/8.11.6/SuSE Linux 0.5) with ESMTP id j69GmFx11662 32, 19 -- for {Microscopy-at-microscopy.com} ; Sat, 9 Jul 2005 10:48:15 -0600 32, 19 -- Received: by hq-dc2.soft-imaging.net with Internet Mail Service (5.5.2657.72) 32, 19 -- id {3GTHZKC1} ; Sat, 9 Jul 2005 10:48:31 -0600 32, 19 -- Message-ID: {6127CE87B9BDD511B59D0001028A497D01189171-at-hq-dc2.soft-imaging.net} 32, 19 -- From: Mike Bode {Mike.Bode-at-soft-imaging.net} 32, 19 -- To: Microscopy-at-microscopy.com 32, 19 -- Subject: RE: [Microscopy] RE: RE: RE: RE: RE: RE: Image review: Ha 32, 19 -- rd copy vs. PC 32, 19 -- Date: Sat, 9 Jul 2005 10:47:25 -0600 32, 19 -- Deferred-Delivery: Sat, 9 Jul 2005 10:47:00 -0600 32, 19 -- MIME-Version: 1.0 32, 19 -- X-Mailer: Internet Mail Service (5.5.2657.72) 32, 19 -- Content-Type: text/plain ==============================End of - Headers==============================
For someone that also has far too much monitor/eye neck strain I also work at 75 cm distance. To make matters worse my desktop systems are both 3200x1200 pixels... (Dual HR Monitors) and I sometimes run with multiple virtual desktops (giving me an effective desktop area of 6400x3200 pixels. You can't display this all at once, but switching is very quick (on a Mac or Linux box)
I guess this is just a comment on technology. We are now overloaded sometimes with far too much information or we are just "parallell processing" alot now adays. Thinking back I don't see how I used to get along with the old 640x480 monitors or even worse the old VT100 text terminals.
Sigh....
Nestor Your Friendly Neighborhood SysOp
==============================Original Headers============================== 5, 11 -- From zaluzec-at-microscopy.com Sat Jul 9 12:04:43 2005 5, 11 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) 5, 11 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j69H4gbk016217 5, 11 -- for {microscopy-at-microscopy.com} ; Sat, 9 Jul 2005 12:04:43 -0500 5, 11 -- Mime-Version: 1.0 5, 11 -- Message-Id: {p06110401bef5b13ba145-at-[206.69.208.22]} 5, 11 -- Date: Sat, 9 Jul 2005 12:04:42 -0500 5, 11 -- To: microscopy-at-microscopy.com 5, 11 -- From: "Nestor J. Zaluzec" {zaluzec-at-microscopy.com} 5, 11 -- Subject: Distance to the Monitor 5, 11 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (normzoo-at-yahoo.com) from http://microscopy.com/MLFormMail.html on Sunday, July 10, 2005 at 04:46:26 ---------------------------------------------------------------------------
Email: normzoo-at-yahoo.com Name: Norman Shedlo
Title-Subject: [Filtered] MListserver: LM Nikon AFM microscope camera
Question: This piece of equipment seems to be relatively common. What is the usual reason for the Nikon AFM microflex shutter to stop working on this model of microscope camera?
The controller works fine and has fresh batteries.
If you can crush, fracture, and polish samples, you are well on your way to preparing samples by other means. South Bay Technology, as well as our competitors, sell complete lines of sample preparation equipment for doing what you want to do. A lot of the expertise of sample prep has been built into the instruments or is contained in the operating manuals. At the South Bay Technology web site, www.southbaytech.com, you can find a list of application notes and technical papers associated with each instrument. At our competitors' sites, you can find similar information. South Bay Technology isn't the only company that has an experienced microscopist on staff (just the best, LOL -Sorry Wolfgang and Rocco. I just couldn't resist.). There are also short courses that involve sample preparation. Ron Anderson and I are teaching a one day short course on TEM sample preparation at the M&M 2005 meeting that you might be interested in attending. We, as well as our competitors, periodically announce short courses on general EM preparation techniques and on specific techniques.
For your immediate needs, I would recommend the MicroCleave(TM) technique, (aka Small Angle Cleavage Technique). This is our model 520 kit and there is a "how to" instruction guide that I wrote that you can get at our site to learn how to do this. It is applicable to silicon, III-V and II-V compounds, SiC, sapphire, GaN, glass, and other brittle materials. It produces extremely good samples and is fairly easy to learn how to do and to teach students and doesn't require any more skills that you have already stated that you have.
For using other sample prep techniques such as dimpling, ion milling, and Tripod Polishing, you can also find specific information about these techniques at our website or you can refer to the four MRS proceedings on TEM Preparation for the Physical Sciences. The common editor in these four proceedings is Ron Anderson. I am not in my office at the moment, or I would give you the volume numbers, but I do remember three: 115, 254, and 480.
Another good way of finding out about these techniques is to find a university with an electron microscopy center that has an experienced lab director and ask for their advice. A few universities that immediately come to mind since I visited or used their facilities are Carnegie Mellon Univ, North Carolina State Univ, Ohio State Univ, Univ of Florida, Univ of California at Irvine, and Penn State Univ. I hope that I haven't slighted anyone. The people at these centers have dealt with a multitude of materials and sample preparation techniques and have a wealth of information. If they can't help you, they will know who can.
I hope this helps.
-Scott
Scott D. Walck, Ph.D. Technical Director South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
I am not familiar with this unit. However, your description sounds like there is a short in the electrodes. Have you checked to see whether evaporated material has put a conductive path between your electrodes? Try putting a current through without the carbon rod to see if you still get a high or even a low current draw. If you do, clean the system.
-Scott
Scott D. Walck, Ph.D. Technical Director South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
-------- Original Message -------- } From: cgarber-at-2spi.com } Sent: Saturday, July 09, 2005 4:14 AM } To: Walck-at-SouthBayTech.com } Subject: [Microscopy] carbon evaporation problem } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Oldrich Benada wrote: } ================================================================= } We have troubles with evaporation control unit BSV 202 of our BALZERS BAF } 301 FE device. Does anybody have some experiences with it? } } The current needed for carbon evaporation suddenly jumped so high that the } 8 A fuse blown out happend. After replacing it, the current needed for } carbon evaporation stays too high and exceeds the permitted current of } safeguarding 8A fuse. There are no differences if we use evaporator 1 } (transformer 1) or evaporator 2 (transformer 2). } For carbon evaporation we use sharpenned carbon rods. } } Thanking in advance for any suggestion.
==============================Original Headers============================== 5, 23 -- From walck-at-southbaytech.com Mon Jul 11 02:15:11 2005 5, 23 -- Received: from smtp05.safesecureweb.com (smtp05.safesecureweb.com [209.41.179.8]) 5, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6B7FBEP002734 5, 23 -- for {microscopy-at-microscopy.com} ; Mon, 11 Jul 2005 02:15:11 -0500 5, 23 -- Received: from localhost (localhost.localdomain [127.0.0.1]) 5, 23 -- by smtp05.safesecureweb.com (Postfix) with ESMTP id 445AE55436F 5, 23 -- for {microscopy-at-microscopy.com} ; Mon, 11 Jul 2005 03:15:11 -0400 (EDT) 5, 23 -- Received: from mail15.safesecureweb.com (unknown [192.168.2.180]) 5, 23 -- by smtp05.safesecureweb.com (Postfix) with ESMTP id C287A55430C 5, 23 -- for {microscopy-at-microscopy.com} ; Mon, 11 Jul 2005 03:15:09 -0400 (EDT) 5, 23 -- MIME-Version: 1.0 5, 23 -- Date: Mon, 11 Jul 2005 03:12:28 -0400 5, 23 -- Content-Type: text/plain; 5, 23 -- charset=iso-8859-1 5, 23 -- Subject: re: [Microscopy] carbon evaporation problem 5, 23 -- From: "Scott Walck" {walck-at-southbaytech.com} 5, 23 -- Reply-To: Walck-at-southbaytech.com 5, 23 -- To: {microscopy-at-microscopy.com} 5, 23 -- Cc: 5, 23 -- Message-ID: {e36e4cc2faa54b4595470c6effbd0eb8-at-southbaytech.com} 5, 23 -- X-Virus-Scanned: by amavisd-new-20030616-p10 at safesecureweb.com 5, 23 -- Content-Transfer-Encoding: 8bit 5, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id j6B7FBEP002734 ==============================End of - Headers==============================
Applicants are being sought for the position of Specialist Light Microscopist {jobs/SeniorMicro.htm} in the Imaging Technology Group (ITG) at the Beckman Institute at the University of Illinois at Urbana-Champaign.
Information regarding this position can be found here: http://www.itg.uiuc.edu/jobs/SeniorMicro.htm
More information on what we do at the Imaging Technology Group can be found here: http://www.itg.uiuc.edu/
Sincerely,
Jonathan M. Ekman Beckman Institute for Advanced Science and Technology, Imaging Technology Group University of Illinois at Urbana-Champaign 405 N. Mathews Avenue Urbana, IL 61801 USA Tel: 217-244-6292 Fax: 217-244-6219
==============================Original Headers============================== 6, 18 -- From ekman-at-itg.uiuc.edu Mon Jul 11 10:17:15 2005 6, 18 -- Received: from zeus.itg.uiuc.edu (zeus.itg.uiuc.edu [130.126.126.162]) 6, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6BFHFhZ019469 6, 18 -- for {Microscopy-at-Microscopy.Com} ; Mon, 11 Jul 2005 10:17:15 -0500 6, 18 -- Received: from [130.126.126.232] (rollins.itg.uiuc.edu [130.126.126.232]) 6, 18 -- by zeus.itg.uiuc.edu (8.12.11/8.12.11) with ESMTP id j6BFHELO020386 6, 18 -- for {Microscopy-at-Microscopy.Com} ; Mon, 11 Jul 2005 10:17:15 -0500 6, 18 -- Message-ID: {42D28D7A.4070300-at-itg.uiuc.edu} 6, 18 -- Date: Mon, 11 Jul 2005 10:17:14 -0500 6, 18 -- From: Jon Ekman {ekman-at-itg.uiuc.edu} 6, 18 -- User-Agent: Mozilla Thunderbird 1.0.2 (Windows/20050317) 6, 18 -- X-Accept-Language: en-us, en 6, 18 -- MIME-Version: 1.0 6, 18 -- To: Microscopy-at-Microscopy.Com 6, 18 -- Subject: LM- Light Microscopy Position - Beckman Institute at the University 6, 18 -- of Illinois at Urbana-Champaign 6, 18 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 6, 18 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (snydert-at-mcmaster.ca) from http://www.microscopy.org/MicroscopyListserver/MLFormMail.html on Monday, July 11, 2005 at 12:42:58 ---------------------------------------------------------------------------
Email: snydert-at-mcmaster.ca Name: Tom Snyder
Organization: McMaster University
Title-Subject: [Filtered] MListserver:JEOL-10S SAM Documentation
Question: Hello,
We are trying to get this Auger microscope up and running but are missing documentation about electron-optics and how they should be set. Specifically we are having trouble locating the beam.
If anyone has access to the documentation or knows what the settings should be I would appreciate any feedback. Also if you know of a good test to verify the opticas are working, that would be great info as well.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (svanhorn-at-notes.cc.sunysb.edu) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, July 11, 2005 at 15:01:44 ---------------------------------------------------------------------------
Email: svanhorn-at-notes.cc.sunysb.edu Name: Sue Van Horn
Organization: SUNY-at-StonyBrook
Education: Graduate College
Location: Stony Brook,NY,USA
Question: has anyone ever used DAP Weldwood contact cement mixed with xylene to help in picking up serial sections???..if so how did you use it and at what dilution??? thanks sue
Sue, I don't remember if it was contact cement or some other glue, but I do remember reading (years ago) about painting the top and bottom sides of the block with a dilute "stick-um" so that as the sections were cut, they adhered to one another via the layer of sticky stuff on the edges. I never tried it, and I don't know how, if the sections adhere so well, one is supposed to separate the ribbon into pieces that will fit on a grid. I know this isn't much help, but at least you know you're not the only one who has heard of such an approach. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
Sue, The use of Weldwood cement "diluted with one or two parts of commercially available contact cement thinner" (20% n-butyl acetate, 80% mineral spirits) to coat the leading and trailing block faces plus many other technique tips for serial sectioning was published by WH Fahrenbach J. Electron Microscopy Technique 1:387-398; 1984. See also EC Henry Stain Technol. 52: 59-60; 1977 who I think was first to use contact cement for this purpose
Iain Miller Department of Biological Sciences Ohio University Athens, OH 45701
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (chrisjohnrhodes-at-hotmail.com) from on Tuesday, July 12, 2005 at 10:56:47 ---------------------------------------------------------------------------
Email: chrisjohnrhodes-at-hotmail.com Name: Chris Rhodes
Organization: Syracuse University
Title-Subject: [Filtered] MListserver:
Question: I'm looking for a semi-permanent mounting medium with the best possible refractive index match to immersion oil and a cover slip (ie as close as possible to 1.515).
The best I have found so far is 1.539
It is for mounting small (less than 2mm) insect structures.
It will be used for CLSM and thus would hopefully have minimal fluorescence.
Due to numerous camera jams over the years we are a bit short on film cassettes for the Philips CM-10 and CM-100. I would appreciate your contacting me if any of you have extra cassettes that you are willing to give/sell to me.
Thanks, Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
I got quite a few from our Philips 300 before we got rid of it. How many do you need? I use them to replace bad ones in our CM10 and they work fine.
Mike
Michael P. Goheen Electron Microscopy Lab Dept. of Pathology & Lab Medicine Indiana University School of Medicine Tel. 317-274-7604 Fax 317-274-5346 mgoheen-at-iupui.edu
-----Original Message----- X-from: dsherman-at-purdue.edu [mailto:dsherman-at-purdue.edu] Sent: Tuesday, July 12, 2005 12:04 PM To: Goheen, Michael P.
Folks,
Due to numerous camera jams over the years we are a bit short on film cassettes for the Philips CM-10 and CM-100. I would appreciate your contacting me if any of you have extra cassettes that you are willing to give/sell to me.
Thanks, Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
I have another one of my "back to the basics" questions that probably make people wonder how the devil I ever got into this business in the first place, but here goes anyway.
We have a client who prepares his own blocks and brings them to us for sectioning, staining, viewing, etc. One day he came into the lab with a bunch of sections on copper mesh grids which he had done immunocytochemistry with standard gold-conjugated secondaries. After viewing them, I offered him the standard advice that next time we would mount some sections on nickel or gold grids if he let us know in advance that he would be doing immunocytochemistry. He said he always used copper and asked why he should switch. My answer was something like "Umm-uhhh.....because it's always done that way" with an additional mumble about copper interfering with the labeling reaction somehow.
Now I have another client, also infinitely more savvy in chemistry that I am, asking the same question.
So I checked through our little in-house research library (again) and did some targeted Googling (again) and found that some folks say that copper: 1) reacts with Tris-HCL buffer (okay, but we hardly ever use that anyway); 2) may react with PBS buffers to produce fine precipitates (but, but....doesn't that mean we should NEVER use Cu with PBS?); 3) reacts with gold colloid (no other explanation given), 4) can alter the charge distribution of the section and cause non-specific background label; and 5) may be oxidized during labelling or interfere with oxidation of chemical groups in the tissue to be labelled. Mostly people just say to use Ni or Au without stating any reasons.
Also, a glance through the literature shows that it's not uncommon to find perfectly successful immunolabelling done on copper grids.
Now, if I were determined to invade a country called Copper any or all of the above reasons could be used with little further explanation, but I'd like to know the Truth and pass it along to my admiring customers.
Any takers?
Thanks in advance.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
==============================Original Headers============================== 15, 23 -- From TindallR-at-missouri.edu Tue Jul 12 17:12:03 2005 15, 23 -- Received: from um-exproto9.um.umsystem.edu (um-exproto9.um.umsystem.edu [207.160.151.49]) 15, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6CMC2XX014275 15, 23 -- for {microscopy-at-microscopy.com} ; Tue, 12 Jul 2005 17:12:03 -0500 15, 23 -- Received: from UM-EMAIL09.um.umsystem.edu ([207.160.151.31]) by um-exproto9.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 15, 23 -- Tue, 12 Jul 2005 17:11:56 -0500 15, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 15, 23 -- Content-class: urn:content-classes:message 15, 23 -- MIME-Version: 1.0 15, 23 -- Content-Type: text/plain; 15, 23 -- charset="us-ascii" 15, 23 -- Subject: TEM: Immunocytochemistry and Cu grids 15, 23 -- Date: Tue, 12 Jul 2005 17:11:47 -0500 15, 23 -- Message-ID: {BA876152E8653240BE8572E897083EE79802F3-at-UM-EMAIL09.um.umsystem.edu} 15, 23 -- X-MS-Has-Attach: 15, 23 -- X-MS-TNEF-Correlator: 15, 23 -- Thread-Topic: TEM: Immunocytochemistry and Cu grids 15, 23 -- Thread-Index: AcWHLrKQfT52UG1vRkSz6rcfHcA8yA== 15, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 15, 23 -- To: {microscopy-at-microscopy.com} 15, 23 -- X-OriginalArrivalTime: 12 Jul 2005 22:11:56.0360 (UTC) FILETIME=[B74F4C80:01C5872E] 15, 23 -- Content-Transfer-Encoding: 8bit 15, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id j6CMC2XX014275 ==============================End of - Headers==============================
I only ran into the "use only Ni or Au grids" rule here, in my current job, and my predecessor certainly produced some superb images showing gold labelling of TEM sections. Blissfully unaware of this rule, I had been using copper grids for all EM immunolabelling. To get good labelling of one particular structure, which is about 40 nm diameter, I used uncoated thin-bar Cu grids so I could get labelling on both sides of the section - seemed to work just fine. I'll be interested to see the responses to your question.
cheers, Rosemary
Dr. Rosemary White rosemary.white-at-csiro.au Microscopy Centre ph. 61-2-6246 5475 CSIRO Plant Industry mob. 61-0402 835 973 GPO Box 1600 fax. 61-2-6246 5334 Canberra, ACT 2601 Australia
} From: TindallR-at-missouri.edu } Reply-To: microscopy-at-microscopy.com } Date: Tue, 12 Jul 2005 17:15:38 -0500 } To: rosemary.white-at-csiro.au } Subject: [Microscopy] TEM: Immunocytochemistry and Cu grids } } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Dear Listers, } } I have another one of my "back to the basics" questions that probably } make people wonder how the devil I ever got into this business in the } first place, but here goes anyway. } } We have a client who prepares his own blocks and brings them to us for } sectioning, staining, viewing, etc. One day he came into the lab with a } bunch of sections on copper mesh grids which he had done } immunocytochemistry with standard gold-conjugated secondaries. After } viewing them, I offered him the standard advice that next time we would } mount some sections on nickel or gold grids if he let us know in advance } that he would be doing immunocytochemistry. He said he always used } copper and asked why he should switch. My answer was something like } "Umm-uhhh.....because it's always done that way" with an additional } mumble about copper interfering with the labeling reaction somehow. } } Now I have another client, also infinitely more savvy in chemistry that } I am, asking the same question. } } So I checked through our little in-house research library (again) and } did some targeted Googling (again) and found that some folks say that } copper: 1) reacts with Tris-HCL buffer (okay, but we hardly ever use } that anyway); 2) may react with PBS buffers to produce fine } precipitates (but, but....doesn't that mean we should NEVER use Cu with } PBS?); 3) reacts with gold colloid (no other explanation given), 4) } can alter the charge distribution of the section and cause non-specific } background label; and 5) may be oxidized during labelling or interfere } with oxidation of chemical groups in the tissue to be labelled. Mostly } people just say to use Ni or Au without stating any reasons. } } Also, a glance through the literature shows that it's not uncommon to } find perfectly successful immunolabelling done on copper grids. } } Now, if I were determined to invade a country called Copper any or all } of the above reasons could be used with little further explanation, but } I'd like to know the Truth and pass it along to my admiring customers. } } Any takers? } } Thanks in advance. } } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility---We Do Small Well! } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-2227 } Email: tindallr-at-missouri.edu } Web: http://www.emc.missouri.edu } } } } } } } } ==============================Original Headers============================== } 15, 23 -- From TindallR-at-missouri.edu Tue Jul 12 17:12:03 2005 } 15, 23 -- Received: from um-exproto9.um.umsystem.edu } (um-exproto9.um.umsystem.edu [207.160.151.49]) } 15, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6CMC2XX014275 } 15, 23 -- for {microscopy-at-microscopy.com} ; Tue, 12 Jul 2005 17:12:03 -0500 } 15, 23 -- Received: from UM-EMAIL09.um.umsystem.edu ([207.160.151.31]) by } um-exproto9.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); } 15, 23 -- Tue, 12 Jul 2005 17:11:56 -0500 } 15, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 } 15, 23 -- Content-class: urn:content-classes:message } 15, 23 -- MIME-Version: 1.0 } 15, 23 -- Content-Type: text/plain; } 15, 23 -- charset="us-ascii" } 15, 23 -- Subject: TEM: Immunocytochemistry and Cu grids } 15, 23 -- Date: Tue, 12 Jul 2005 17:11:47 -0500 } 15, 23 -- Message-ID: } {BA876152E8653240BE8572E897083EE79802F3-at-UM-EMAIL09.um.umsystem.edu} } 15, 23 -- X-MS-Has-Attach: } 15, 23 -- X-MS-TNEF-Correlator: } 15, 23 -- Thread-Topic: TEM: Immunocytochemistry and Cu grids } 15, 23 -- Thread-Index: AcWHLrKQfT52UG1vRkSz6rcfHcA8yA== } 15, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} } 15, 23 -- To: {microscopy-at-microscopy.com} } 15, 23 -- X-OriginalArrivalTime: 12 Jul 2005 22:11:56.0360 (UTC) } FILETIME=[B74F4C80:01C5872E] } 15, 23 -- Content-Transfer-Encoding: 8bit } 15, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } ns.microscopy.com id j6CMC2XX014275 } ==============================End of - Headers============================== }
==============================Original Headers============================== 6, 23 -- From Rosemary.White-at-csiro.au Tue Jul 12 17:53:34 2005 6, 23 -- Received: from vic-ironport-ext-out2.csiro.au (vic-ironport-ext-out2.csiro.au [150.229.64.38]) 6, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6CMrXDw022373 6, 23 -- for {microscopy-at-microscopy.com} ; Tue, 12 Jul 2005 17:53:34 -0500 6, 23 -- Received: from exgw1-cbr.nexus.csiro.au (152.83.3.66) 6, 23 -- by vic-ironport-ext-out2.csiro.au with ESMTP; 13 Jul 2005 08:53:32 +1000 6, 23 -- X-BrightmailFiltered: true 6, 23 -- X-Brightmail-Tracker: AAAAAQAAA+k= 6, 23 -- X-IronPort-AV: i="3.93,284,1114956000"; 6, 23 -- d="scan'208"; a="45339190:sNHT23254752" 6, 23 -- Received: from [152.83.167.45] ([152.83.167.45]) by exgw1-cbr.nexus.csiro.au with Microsoft SMTPSVC(5.0.2195.6713); 6, 23 -- Wed, 13 Jul 2005 08:53:31 +1000 6, 23 -- User-Agent: Microsoft-Entourage/10.1.4.030702.0 6, 23 -- Date: Wed, 13 Jul 2005 08:54:42 +1000 6, 23 -- Subject: Re: [Microscopy] TEM: Immunocytochemistry and Cu grids 6, 23 -- From: Rosemary White {Rosemary.White-at-csiro.au} 6, 23 -- To: {microscopy-at-microscopy.com} 6, 23 -- Message-ID: {BEFA8752.108B7%Rosemary.White-at-csiro.au} 6, 23 -- In-Reply-To: {200507122215.j6CMFcFZ019486-at-ns.microscopy.com} 6, 23 -- Mime-version: 1.0 6, 23 -- Content-type: text/plain; charset="US-ASCII" 6, 23 -- Content-transfer-encoding: 7bit 6, 23 -- X-OriginalArrivalTime: 12 Jul 2005 22:53:31.0241 (UTC) FILETIME=[865FED90:01C58734] ==============================End of - Headers==============================
As usual, I am hoping to enlist the vast knowledge and experience of the this group to solve a perplexing problem. Here is the puzzle.
We have noticed artifact elemental peaks in EDS spectra obtained with our Oxford Inca system on a Hitachi S-4700 FESEM. For example, an analysis in the center of a strip of 6 mm wide clean copper tape on an aluminum stub generates a spectrum with large peaks for copper (this is good) and a small aluminum peak (not good). Repeat the experiment with a carbon stub, and the small aluminum peak is replaced by a carbon peak.
Oddly, the phenomenon is observed only with the microscope operating in High Mag mode. The same analysis (same mag, count rate, etc.) on the copper tape in Low Mag mode detects only copper. This result is similar to what we might expect from beam scatter in a variable pressure SEM.
Anyone one out there experienced this problem? Anyone with a similar microscope willing to try the copper tape experiment to let us know if this is unique to our instrument or fundamental to the S-4700. Hitachi and Oxford are perplexed so far.
Thanks for your kind assistance.
-- Larry D. Hanke, P.E. Materials Evaluation and Engineering, Inc. Practical Solutions Through Technology and Innovation http://www.mee-inc.com (763) 449-8870
==============================Original Headers============================== 8, 22 -- From hanke-at-mee-inc.com Tue Jul 12 18:02:48 2005 8, 22 -- Received: from mail5.atl.registeredsite.com (mail5.atl.registeredsite.com [64.224.219.79]) 8, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6CN2mBT030143 8, 22 -- for {microscopy-at-microscopy.com} ; Tue, 12 Jul 2005 18:02:48 -0500 8, 22 -- Received: from netmail.mail.registeredsite.com ([216.122.69.14]) 8, 22 -- by mail5.atl.registeredsite.com (8.12.11/8.12.8) with ESMTP id j6CN2mDA007877 8, 22 -- for {microscopy-at-microscopy.com} ; Tue, 12 Jul 2005 23:02:48 GMT 8, 22 -- Received: (qmail 61036 invoked by uid 89); 12 Jul 2005 23:09:35 -0000 8, 22 -- Received: from unknown (HELO ?192.168.1.109?) (216.43.123.204) 8, 22 -- by mail.billdalton2002.com with SMTP; 12 Jul 2005 23:09:35 -0000 8, 22 -- Message-ID: {42D44C17.4030804-at-mee-inc.com} 8, 22 -- Date: Tue, 12 Jul 2005 18:02:47 -0500 8, 22 -- From: Larry Hanke {hanke-at-mee-inc.com} 8, 22 -- User-Agent: Mozilla Thunderbird 1.0.2 (Windows/20050317) 8, 22 -- X-Accept-Language: en-us, en 8, 22 -- MIME-Version: 1.0 8, 22 -- To: microscopy-at-microscopy.com 8, 22 -- Subject: Stray X-rays in FE SEM? 8, 22 -- References: {200507122213.j6CMDHwb015857-at-ns.microscopy.com} 8, 22 -- In-Reply-To: {200507122213.j6CMDHwb015857-at-ns.microscopy.com} 8, 22 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 8, 22 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Hi Randy, I had problems with Cu grids when incubating sections for long periods - overnight for instance. The copper would react with whatever was available and form green-blue salts. Not unexpected. We switched to nickel grids for a while but they charge, gold but they are delicate...by this time we had shortened the incubation times used, and tried copper again... And had no problem using coated grids and incubation times of an hour or so. Grids are floated on drops of media so that only the coated side is wetted - I don't know if that is important.
Dr SJ Stowe Facility Coordinator ANU Electron Microscopy Unit ANU CRICOS#00120C
} -----Original Message----- } From: TindallR-at-missouri.edu [mailto:TindallR-at-missouri.edu] } Sent: Wednesday, 13 July 2005 8:12 AM } To: sally.stowe-at-anu.edu.au } Subject: [Microscopy] TEM: Immunocytochemistry and Cu grids } } } } } } --------------------------------------------------------------- } ------------- } The Microscopy ListServer -- CoSponsor: The Microscopy } Society of America To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver
==============================Original Headers============================== 7, 27 -- From sally.stowe-at-anu.edu.au Tue Jul 12 18:07:03 2005 7, 27 -- Received: from mail.rsbs.anu.edu.au (rsbspc309.anu.edu.au [150.203.72.70]) 7, 27 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6CN729f005441 7, 27 -- for {microscopy-at-microscopy.com} ; Tue, 12 Jul 2005 18:07:03 -0500 7, 27 -- Received: from rsbs2262 (rsbs2262.rsbs.anu.edu.au [150.203.36.144]) 7, 27 -- by mail.rsbs.anu.edu.au (Postfix) with ESMTP id A10A1F4408C 7, 27 -- for {microscopy-at-microscopy.com} ; Wed, 13 Jul 2005 09:06:25 +1000 (EST) 7, 27 -- Reply-To: {Sally.Stowe-at-anu.edu.au} 7, 27 -- From: "Sally Stowe" {Sally.Stowe-at-anu.edu.au} 7, 27 -- To: {microscopy-at-microscopy.com} 7, 27 -- Subject: RE: [Microscopy] TEM: Immunocytochemistry and Cu grids 7, 27 -- Date: Wed, 13 Jul 2005 09:06:26 +1000 7, 27 -- Message-ID: {006201c58736$549b3a10$9024cb96-at-rsbs.anu.edu.au} 7, 27 -- MIME-Version: 1.0 7, 27 -- Content-Type: text/plain; 7, 27 -- charset="US-ASCII" 7, 27 -- X-Priority: 3 (Normal) 7, 27 -- X-MSMail-Priority: Normal 7, 27 -- X-Mailer: Microsoft Outlook, Build 10.0.6626 7, 27 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1409 7, 27 -- In-Reply-To: {200507122212.j6CMCCCH014469-at-ns.microscopy.com} 7, 27 -- Importance: Normal 7, 27 -- X-RSBS-MailScanner-Information: Please contact the ISP for more information 7, 27 -- X-RSBS-MailScanner: Found to be clean 7, 27 -- X-MailScanner-From: sally.stowe-at-anu.edu.au 7, 27 -- Content-Transfer-Encoding: 8bit 7, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id j6CN729f005441 ==============================End of - Headers==============================
What is the analytical WD for EDS in this SEM? What WD are you using?
gary g.
At 04:04 PM 7/12/2005, you wrote:
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==============================Original Headers============================== 8, 18 -- From gary-at-gaugler.com Tue Jul 12 18:30:49 2005 8, 18 -- Received: from smtp2.mc.surewest.net (smtp2.mc.surewest.net [66.60.130.51]) 8, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id j6CNUn3P013376 8, 18 -- for {microscopy-at-microscopy.com} ; Tue, 12 Jul 2005 18:30:49 -0500 8, 18 -- Received: (s3-16861); Tue, 12 Jul 2005 16:30:48 -0700 8, 18 -- Received: from unknown (66.60.171.211) 8, 18 -- by smtp2.mc.surewest.net (s3-smtpd/0.90-beta3) with SMTP; Tue, 12 Jul 2005 16:30:47 -0700 8, 18 -- Message-Id: {6.2.0.14.2.20050712163005.02861a30-at-mail.calweb.com} 8, 18 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.0.14 8, 18 -- Date: Tue, 12 Jul 2005 16:30:46 -0700 8, 18 -- To: microscopy-at-microscopy.com 8, 18 -- From: Gary Gaugler {gary-at-gaugler.com} 8, 18 -- Subject: Re: [Microscopy] Stray X-rays in FE SEM? 8, 18 -- In-Reply-To: {200507122304.j6CN4K2C001228-at-ns.microscopy.com} 8, 18 -- References: {200507122304.j6CN4K2C001228-at-ns.microscopy.com} 8, 18 -- Mime-Version: 1.0 8, 18 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 8, 18 -- X-TST: smtp2.mc.surewest.net SNWK3 0.31-80 ip=66.60.171.211 ==============================End of - Headers==============================
I didn't think the ones from the EM series would work in the CM series. Our EM-400 took the larger plate cassettes with the film inserts. I thought those inserts were different from the film holders that came with the CM series but I may be wrong. If they do work than I may still have a few around as well. I gave away most of my cameras and cassettes from the EM-400 when we decommissioned it but still may have a few around. I'll check and let you know if I would like to try the ones you have.
Are you going to M&M2005?
Debby
On 7/12/05 1:40 PM, "mgoheen-at-iupui.edu" {mgoheen-at-iupui.edu} wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Hi Debby, } } I got quite a few from our Philips 300 before we got rid of it. How many } do you need? I use them to replace bad ones in our CM10 and they work } fine. } } Mike } } Michael P. Goheen } Electron Microscopy Lab } Dept. of Pathology & Lab Medicine } Indiana University School of Medicine } Tel. 317-274-7604 } Fax 317-274-5346 } mgoheen-at-iupui.edu } } } -----Original Message----- } X-from: dsherman-at-purdue.edu [mailto:dsherman-at-purdue.edu] } Sent: Tuesday, July 12, 2005 12:04 PM } To: Goheen, Michael P. } Subject: [Microscopy] Film cassettes } } } } } ------------------------------------------------------------------------ } ---- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ } ---- } } Folks, } } Due to numerous camera jams over the years we are a bit short on film } cassettes for the Philips CM-10 and CM-100. I would appreciate your } contacting me if any of you have extra cassettes that you are willing to } give/sell to me. } } Thanks, } Debby } } Debby Sherman, Manager Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } S-052 Whistler Building } 170 S. University Street } West Lafayette, IN 47907 } http://www3.agriculture.purdue.edu/microscopy } } } } ==============================Original } Headers============================== } 6, 20 -- From dsherman-at-purdue.edu Tue Jul 12 12:00:35 2005 } 6, 20 -- Received: from mailhub248.itcs.purdue.edu } (mailhub248.itcs.purdue.edu [128.210.5.248]) } 6, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id } j6CH0ZKY027403 } 6, 20 -- for {microscopy-at-microscopy.com} ; Tue, 12 Jul 2005 } 12:00:35 -0500 } 6, 20 -- Received: from [192.168.1.104] } (cpe-24-25-218-216.san.res.rr.com [24.25.218.216]) } 6, 20 -- (authenticated bits=0) } 6, 20 -- by mailhub248.itcs.purdue.edu } (8.13.4/8.13.4/avscan-smtp) with ESMTP id j6CH0VTF031403 } 6, 20 -- for {microscopy-at-microscopy.com} ; Tue, 12 Jul 2005 } 12:00:33 -0500 } 6, 20 -- User-Agent: Microsoft-Entourage/11.0.0.040405 } 6, 20 -- Date: Tue, 12 Jul 2005 12:00:29 -0500 } 6, 20 -- Subject: Film cassettes } 6, 20 -- From: Debby Sherman {dsherman-at-purdue.edu} } 6, 20 -- To: "message to: MSA list" {microscopy-at-microscopy.com} } 6, 20 -- Message-ID: {BEF9615D.4A27%dsherman-at-purdue.edu} } 6, 20 -- Mime-version: 1.0 } 6, 20 -- Content-type: text/plain; } 6, 20 -- charset="US-ASCII" } 6, 20 -- Content-transfer-encoding: 7bit } 6, 20 -- X-PMX-Version: 4.7.1.128075 } 6, 20 -- X-PerlMx-Virus-Scanned: Yes } ==============================End of - } Headers============================== } } } ==============================Original Headers============================== } 17, 26 -- From mgoheen-at-iupui.edu Tue Jul 12 13:37:06 2005 } 17, 26 -- Received: from julesburg.uits.indiana.edu } (julesburg.uits.indiana.edu [129.79.1.75]) } 17, 26 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6CIb6X8003989 } 17, 26 -- for {microscopy-at-microscopy.com} ; Tue, 12 Jul 2005 13:37:06 -0500 } 17, 26 -- Received: from iu-mssg-smtp03.ads.iu.edu } (iu-mssg-smtp03.exchange.iu.edu [129.79.1.220]) } 17, 26 -- by julesburg.uits.indiana.edu (8.12.10/8.12.10/IUPO) with ESMTP id } j6CIb4En026636 } 17, 26 -- for {microscopy-at-microscopy.com} ; Tue, 12 Jul 2005 13:37:04 -0500 } (EST) } 17, 26 -- Received: from iu-mssg-mbx01.ads.iu.edu ([129.79.1.210]) by } iu-mssg-smtp03.ads.iu.edu with Microsoft SMTPSVC(5.0.2195.6713); } 17, 26 -- Tue, 12 Jul 2005 13:37:04 -0500 } 17, 26 -- X-MIMEOLE: Produced By Microsoft Exchange V6.0.6603.0 } 17, 26 -- content-class: urn:content-classes:message } 17, 26 -- MIME-Version: 1.0 } 17, 26 -- Content-Type: text/plain; } 17, 26 -- charset="us-ascii" } 17, 26 -- Subject: RE: [Microscopy] Film cassettes } 17, 26 -- Date: Tue, 12 Jul 2005 13:37:04 -0500 } 17, 26 -- Message-ID: } {CED839BBC1F79B439C630BEBD9661A6A010CAB24-at-iu-mssg-mbx01.exchange.iu.edu} } 17, 26 -- X-MS-Has-Attach: } 17, 26 -- X-MS-TNEF-Correlator: } 17, 26 -- Thread-Topic: [Microscopy] Film cassettes } 17, 26 -- Thread-Index: AcWHA7eWMUED3onrTPq+k9A7IC4dxgADHRjg } 17, 26 -- From: "Goheen, Michael P." {mgoheen-at-iupui.edu} } 17, 26 -- To: {microscopy-at-microscopy.com} } 17, 26 -- X-OriginalArrivalTime: 12 Jul 2005 18:37:04.0891 (UTC) } FILETIME=[B3653CB0:01C58710] } 17, 26 -- Content-Transfer-Encoding: 8bit } 17, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } ns.microscopy.com id j6CIb6X8003989 } ==============================End of - Headers==============================
I'm not overly familiar with the Hitachi SEMs, but is the "hi-mag" mode on your SEM an immersion lens mode? This is most likely the case if you are using an "in-lens" or "in-column" detector. If so, it is possible that the BSE are being bent around by the immersion field and striking back on the sample. Depending on the immersion field, this could be close or far from the beam. These will probably still be within the viewing angle of your EDS collimator. In their immersion lens SEMs, FEI provides an EDS mode with a weakened immersion lens field so that the BSE are bent somewhat less and strike outside the field of view of the EDS collimator.
Cheers, Henk
At 07:04 PM 7/12/2005, hanke-at-mee-inc.com wrote:
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Hendrik O. Colijn colijn.1-at-osu.edu OSU Campus Electron Optics Facility www.ceof.ohio-state.edu 040 Fontana Labs, 116 W. 19th Ave
==============================Original Headers============================== 10, 25 -- From colijn.1-at-osu.edu Tue Jul 12 19:12:25 2005 10, 25 -- Received: from ER6S1.ENG.OHIO-STATE.EDU (er6s1.ecr6.ohio-state.edu [164.107.76.2]) 10, 25 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6D0CPK8028631 10, 25 -- for {microscopy-at-microscopy.com} ; Tue, 12 Jul 2005 19:12:25 -0500 10, 25 -- Received: from CONVERSION-DAEMON.er6s1.eng.ohio-state.edu by 10, 25 -- er6s1.eng.ohio-state.edu (PMDF V6.2-1x5 #31056) 10, 25 -- id {01LQJTLJP7CW9HC4MN-at-er6s1.eng.ohio-state.edu} for 10, 25 -- microscopy-at-microscopy.com; Tue, 12 Jul 2005 20:12:24 -0400 (EDT) 10, 25 -- Received: from HOC3.osu.edu 10, 25 -- (d149-67-33-174.col.wideopenwest.com [67.149.174.33]) 10, 25 -- by er6s1.eng.ohio-state.edu (PMDF V6.2-1x5 #31056) 10, 25 -- with ESMTPA id {01LQJTLISNP09H2SL1-at-er6s1.eng.ohio-state.edu} for 10, 25 -- microscopy-at-microscopy.com; Tue, 12 Jul 2005 20:12:24 -0400 (EDT) 10, 25 -- Date: Tue, 12 Jul 2005 20:12:10 -0400 10, 25 -- From: "Hendrik O. Colijn" {colijn.1-at-osu.edu} 10, 25 -- Subject: Re: [Microscopy] Stray X-rays in FE SEM? 10, 25 -- In-reply-to: {200507122304.j6CN4Fdd000963-at-ns.microscopy.com} 10, 25 -- Sender: colijn-at-er6s1.ecr6.ohio-state.edu 10, 25 -- To: microscopy-at-microscopy.com 10, 25 -- Message-id: {6.2.0.14.2.20050712200205.01ea9018-at-mail.er6.eng.ohio-state.edu} 10, 25 -- MIME-version: 1.0 10, 25 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.0.14 10, 25 -- Content-type: text/plain; charset=us-ascii; format=flowed 10, 25 -- X-Env-From: auth/colijn.1-at-osu.edu 10, 25 -- References: {200507122304.j6CN4Fdd000963-at-ns.microscopy.com} ==============================End of - Headers==============================
The CM series used both the 36 and 56 sheet film boxes. We have both an early CM12 which used the 2-part film carriers (36 exposure box; same as our old EM400) and a CM200 with the thinner single piece film carriers (56 exposure box). I don't know if the EM300 used the same film carriers as the EM400 series since the camera boxes in the EM300 we had (years ago) were different. The EM400 and later Philips/FEI scopes use the single "behind the column" film box assembly
I believe the issue is more the film box than the microscope. ...and more specifically, the film transport tray since the 2 carriers are different thicknesses. The boxes have the same outside dimensions and are (I believe) interchangeable. In other words, I could take my old 36 exposure film box, drop it right into my CM200 and only need to change the film stock number. CAUTION -- I haven't actually tested this since I haven't felt motivated to reduce the number of exposures in my microscope!
Cheers, Henk
At 08:10 PM 7/12/2005, dsherman-at-purdue.edu wrote:
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Hendrik O. Colijn colijn.1-at-osu.edu OSU Campus Electron Optics Facility www.ceof.ohio-state.edu 040 Fontana Labs, 116 W. 19th Ave
==============================Original Headers============================== 9, 25 -- From colijn.1-at-osu.edu Tue Jul 12 19:24:11 2005 9, 25 -- Received: from ER6S1.ENG.OHIO-STATE.EDU (er6s1.ecr6.ohio-state.edu [164.107.76.2]) 9, 25 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6D0OB9s004623 9, 25 -- for {microscopy-at-microscopy.com} ; Tue, 12 Jul 2005 19:24:11 -0500 9, 25 -- Received: from CONVERSION-DAEMON.er6s1.eng.ohio-state.edu by 9, 25 -- er6s1.eng.ohio-state.edu (PMDF V6.2-1x5 #31056) 9, 25 -- id {01LQJU14I6PC9H5VYA-at-er6s1.eng.ohio-state.edu} for 9, 25 -- microscopy-at-microscopy.com; Tue, 12 Jul 2005 20:24:11 -0400 (EDT) 9, 25 -- Received: from HOC3.osu.edu 9, 25 -- (d149-67-33-174.col.wideopenwest.com [67.149.174.33]) 9, 25 -- by er6s1.eng.ohio-state.edu (PMDF V6.2-1x5 #31056) 9, 25 -- with ESMTPA id {01LQJU13LJA89H2SL1-at-er6s1.eng.ohio-state.edu} for 9, 25 -- microscopy-at-microscopy.com; Tue, 12 Jul 2005 20:24:10 -0400 (EDT) 9, 25 -- Date: Tue, 12 Jul 2005 20:24:04 -0400 9, 25 -- From: "Hendrik O. Colijn" {colijn.1-at-osu.edu} 9, 25 -- Subject: Re: [Microscopy] Film cassettes 9, 25 -- In-reply-to: {200507130010.j6D0A1MX024791-at-ns.microscopy.com} 9, 25 -- Sender: colijn-at-er6s1.ecr6.ohio-state.edu 9, 25 -- To: microscopy-at-microscopy.com 9, 25 -- Message-id: {6.2.0.14.2.20050712201248.01eb33d0-at-mail.er6.eng.ohio-state.edu} 9, 25 -- MIME-version: 1.0 9, 25 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.0.14 9, 25 -- Content-type: text/plain; charset=us-ascii; format=flowed 9, 25 -- X-Env-From: auth/colijn.1-at-osu.edu 9, 25 -- References: {200507130010.j6D0A1MX024791-at-ns.microscopy.com} ==============================End of - Headers==============================
We used Cu grids for many years. Normally we use formvar + carbon coated grids for ICC since we have a lot less loss of sections. We also tend to do long incubations ...usually overnight. This is time efficient and also lets us use highly diluted antibody so also minimize background due to cross-reactions from contaminants in polyclonal antibodies.
Occasionally we would have a reaction due to copper oxidizing with resultant green solution. I attributed this to salts or traces of Tween 20 in the buffer and breaks in the coating exposing the Cu. As far as I can see, this is the only reason for not using Cu grids if you use coated grids. We have switched for the most part to using coated Ni grids to avoid this problem.
On the other hand, occasionally we need to do a double labeling using uncoated grids and both sides of the sections. I would hesitate to use Cu for this and prefer Ni. Gold would work but is both more expensive and more delicate than Ni grids.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
On 7/12/05 5:15 PM, "TindallR-at-missouri.edu" {TindallR-at-missouri.edu} wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Dear Listers, } } I have another one of my "back to the basics" questions that probably } make people wonder how the devil I ever got into this business in the } first place, but here goes anyway. } } We have a client who prepares his own blocks and brings them to us for } sectioning, staining, viewing, etc. One day he came into the lab with a } bunch of sections on copper mesh grids which he had done } immunocytochemistry with standard gold-conjugated secondaries. After } viewing them, I offered him the standard advice that next time we would } mount some sections on nickel or gold grids if he let us know in advance } that he would be doing immunocytochemistry. He said he always used } copper and asked why he should switch. My answer was something like } "Umm-uhhh.....because it's always done that way" with an additional } mumble about copper interfering with the labeling reaction somehow. } } Now I have another client, also infinitely more savvy in chemistry that } I am, asking the same question. } } So I checked through our little in-house research library (again) and } did some targeted Googling (again) and found that some folks say that } copper: 1) reacts with Tris-HCL buffer (okay, but we hardly ever use } that anyway); 2) may react with PBS buffers to produce fine } precipitates (but, but....doesn't that mean we should NEVER use Cu with } PBS?); 3) reacts with gold colloid (no other explanation given), 4) } can alter the charge distribution of the section and cause non-specific } background label; and 5) may be oxidized during labelling or interfere } with oxidation of chemical groups in the tissue to be labelled. Mostly } people just say to use Ni or Au without stating any reasons. } } Also, a glance through the literature shows that it's not uncommon to } find perfectly successful immunolabelling done on copper grids. } } Now, if I were determined to invade a country called Copper any or all } of the above reasons could be used with little further explanation, but } I'd like to know the Truth and pass it along to my admiring customers. } } Any takers? } } Thanks in advance. } } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility---We Do Small Well! } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-2227 } Email: tindallr-at-missouri.edu } Web: http://www.emc.missouri.edu } } } } } } } } ==============================Original Headers============================== } 15, 23 -- From TindallR-at-missouri.edu Tue Jul 12 17:12:03 2005 } 15, 23 -- Received: from um-exproto9.um.umsystem.edu } (um-exproto9.um.umsystem.edu [207.160.151.49]) } 15, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6CMC2XX014275 } 15, 23 -- for {microscopy-at-microscopy.com} ; Tue, 12 Jul 2005 17:12:03 -0500 } 15, 23 -- Received: from UM-EMAIL09.um.umsystem.edu ([207.160.151.31]) by } um-exproto9.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); } 15, 23 -- Tue, 12 Jul 2005 17:11:56 -0500 } 15, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 } 15, 23 -- Content-class: urn:content-classes:message } 15, 23 -- MIME-Version: 1.0 } 15, 23 -- Content-Type: text/plain; } 15, 23 -- charset="us-ascii" } 15, 23 -- Subject: TEM: Immunocytochemistry and Cu grids } 15, 23 -- Date: Tue, 12 Jul 2005 17:11:47 -0500 } 15, 23 -- Message-ID: } {BA876152E8653240BE8572E897083EE79802F3-at-UM-EMAIL09.um.umsystem.edu} } 15, 23 -- X-MS-Has-Attach: } 15, 23 -- X-MS-TNEF-Correlator: } 15, 23 -- Thread-Topic: TEM: Immunocytochemistry and Cu grids } 15, 23 -- Thread-Index: AcWHLrKQfT52UG1vRkSz6rcfHcA8yA== } 15, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} } 15, 23 -- To: {microscopy-at-microscopy.com} } 15, 23 -- X-OriginalArrivalTime: 12 Jul 2005 22:11:56.0360 (UTC) } FILETIME=[B74F4C80:01C5872E] } 15, 23 -- Content-Transfer-Encoding: 8bit } 15, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } ns.microscopy.com id j6CMC2XX014275 } ==============================End of - Headers==============================
==============================Original Headers============================== 12, 21 -- From dsherman-at-purdue.edu Tue Jul 12 19:46:33 2005 12, 21 -- Received: from mailhub248.itcs.purdue.edu (mailhub248.itcs.purdue.edu [128.210.5.248]) 12, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6D0kWdK013786 12, 21 -- for {microscopy-at-microscopy.com} ; Tue, 12 Jul 2005 19:46:32 -0500 12, 21 -- Received: from [192.168.1.104] (cpe-24-25-218-216.san.res.rr.com [24.25.218.216]) 12, 21 -- (authenticated bits=0) 12, 21 -- by mailhub248.itcs.purdue.edu (8.13.4/8.13.4/avscan-smtp) with ESMTP id j6D0kUI5025545 12, 21 -- for {microscopy-at-microscopy.com} ; Tue, 12 Jul 2005 19:46:31 -0500 12, 21 -- User-Agent: Microsoft-Entourage/11.0.0.040405 12, 21 -- Date: Tue, 12 Jul 2005 19:46:28 -0500 12, 21 -- Subject: Re: [Microscopy] TEM: Immunocytochemistry and Cu grids 12, 21 -- From: Debby Sherman {dsherman-at-purdue.edu} 12, 21 -- To: "message to: MSA list " {microscopy-at-microscopy.com} 12, 21 -- Message-ID: {BEF9CE94.4A5B%dsherman-at-purdue.edu} 12, 21 -- In-Reply-To: {200507122215.j6CMFI6D018851-at-ns.microscopy.com} 12, 21 -- Mime-version: 1.0 12, 21 -- Content-type: text/plain; 12, 21 -- charset="US-ASCII" 12, 21 -- Content-transfer-encoding: 7bit 12, 21 -- X-PMX-Version: 4.7.1.128075 12, 21 -- X-PerlMx-Virus-Scanned: Yes ==============================End of - Headers==============================
The film holders for the Philips EM400 and the CM series are the same. The same holders can be used in the Philips EM201,EM300, EM301 . With the inserts you load cut film without you load glass plates. ----- Original Message ----- X-from: {dsherman-at-purdue.edu} To: {amtecss-at-earthlink.net} Sent: Tuesday, July 12, 2005 8:08 PM
randy
you don't say what your client's stuff looked like. that would help.
had a former director, before i grew up and went back to school - or was that truly growing up? anyway, out of nickel grids, not to mention formvar-nickel. she wanted IEM done. then! forthwith! no delay! do not put it of! threat of discipline if not in her hands that afternoon!! etc. insisted i use copper. this was also on formvar coated grids. not pretty, quite nasty, large semicrystals of junk all over. really did not look good after the second and third try.
there were several possible problems. first, we spun things onto the grid, 30 minutes in an air driven ultracentrifuge. so the copper back surface was guaranteed to get wet and react with the buffer. or perhaps it was the buffer she had things in, i never could get a straight answer from her on that one.
recently i did a long ultracentrifuge run to load some 10S particles onto a formvar-copper grid. the grids also reacted with the phosphate buffer over the 4 hour spin time. again, not a pretty picture - i had to go back to my collaborator and get fresh material so we could put it onto formvar-nickel grids. wonderful gentleman. much more mature. real pleasure to collaborate with.
having said that, i read what rosemary white, sally stowe and debbie sherman have said. i make no pretense about my opinion on dogma (dogma-schmogma, it ain't gospel until i try it and prove the fact to me personally). give it a go and let the rest of us try out to see what happens. perhaps i will even try it in my overworked 10 finger lab. but after the two molecular experiments i need to do in order get that paper of mine accepted on resubmission, and the 7 other collaborators' projects, the looking at micrographs from 2 outside sources with questions, etc. all right, i exagerate, it's not quite that bad but i really do have to get back to one person about his results before i go on holiday tomorrow.
give it a shot and let us know
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-9354 Cell:204-781-1502 Fax:204-789-3926
==============================Original Headers============================== 10, 18 -- From paul_hazelton-at-umanitoba.ca Tue Jul 12 22:55:26 2005 10, 18 -- Received: from electra.cc.umanitoba.ca (electra.cc.umanitoba.ca [130.179.16.23]) 10, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6D3tQXP031283 10, 18 -- for {microscopy-at-microscopy.com} ; Tue, 12 Jul 2005 22:55:26 -0500 10, 18 -- Received: from umanitoba.ca (cvx-045.cc.umanitoba.ca [130.179.152.108]) 10, 18 -- by electra.cc.umanitoba.ca (8.13.0/8.13.0) with ESMTP id j6D3tH1b004399 10, 18 -- for {microscopy-at-microscopy.com} ; Tue, 12 Jul 2005 22:55:23 -0500 (CDT) 10, 18 -- Message-ID: {42D4924F.EE51DAB5-at-umanitoba.ca} 10, 18 -- Date: Tue, 12 Jul 2005 23:02:30 -0500 10, 18 -- From: paul r hazelton {paul_hazelton-at-umanitoba.ca} 10, 18 -- X-Mailer: Mozilla 4.78 [en] (Win98; U) 10, 18 -- X-Accept-Language: en 10, 18 -- MIME-Version: 1.0 10, 18 -- To: microscopy-at-microscopy.com 10, 18 -- Subject: Re: [Microscopy] TEM: Immunocytochemistry and Cu grids 10, 18 -- References: {200507122215.j6CMF5cr018469-at-ns.microscopy.com} 10, 18 -- Content-Type: text/plain; charset=us-ascii 10, 18 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
With magnetic field active, test stray signal as a function of WD. Same experiment with a charge sensitive sample should also be interesting, not for x-ray, but SE. response.
-----Original Message----- X-from: "hanke-at-mee-inc.com" {hanke-at-mee-inc.com} Sent: Tuesday, July 12, 2005 11:03 PM To: "eprincipe01-at-hotmail.com" {eprincipe01-at-hotmail.com}
Dear Listers:
As usual, I am hoping to enlist the vast knowledge and experience of the this group to solve a perplexing problem. Here is the puzzle.
We have noticed artifact elemental peaks in EDS spectra obtained with our Oxford Inca system on a Hitachi S-4700 FESEM. For example, an analysis in the center of a strip of 6 mm wide clean copper tape on an aluminum stub generates a spectrum with large peaks for copper (this is good) and a small aluminum peak (not good). Repeat the experiment with a carbon stub, and the small aluminum peak is replaced by a carbon peak.
Oddly, the phenomenon is observed only with the microscope operating in High Mag mode. The same analysis (same mag, count rate, etc.) on the copper tape in Low Mag mode detects only copper. This result is similar to what we might expect from beam scatter in a variable pressure SEM.
Anyone one out there experienced this problem? Anyone with a similar microscope willing to try the copper tape experiment to let us know if this is unique to our instrument or fundamental to the S-4700. Hitachi and Oxford are perplexed so far.
Thanks for your kind assistance.
-- Larry D. Hanke, P.E. Materials Evaluation and Engineering, Inc. Practical Solutions Through Technology and Innovation http://www.mee-inc.com (763) 449-8870
==============================Original Headers============================== 8, 22 -- From hanke-at-mee-inc.com Tue Jul 12 18:02:48 2005 8, 22 -- Received: from mail5.atl.registeredsite.com (mail5.atl.registeredsite.com [64.224.219.79]) 8, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6CN2mBT030143 8, 22 -- for {microscopy-at-microscopy.com} ; Tue, 12 Jul 2005 18:02:48 -0500 8, 22 -- Received: from netmail.mail.registeredsite.com ([216.122.69.14]) 8, 22 -- by mail5.atl.registeredsite.com (8.12.11/8.12.8) with ESMTP id j6CN2mDA007877 8, 22 -- for {microscopy-at-microscopy.com} ; Tue, 12 Jul 2005 23:02:48 GMT 8, 22 -- Received: (qmail 61036 invoked by uid 89); 12 Jul 2005 23:09:35 -0000 8, 22 -- Received: from unknown (HELO ?192.168.1.109?) (216.43.123.204) 8, 22 -- by mail.billdalton2002.com with SMTP; 12 Jul 2005 23:09:35 -0000 8, 22 -- Message-ID: {42D44C17.4030804-at-mee-inc.com} 8, 22 -- Date: Tue, 12 Jul 2005 18:02:47 -0500 8, 22 -- From: Larry Hanke {hanke-at-mee-inc.com} 8, 22 -- User-Agent: Mozilla Thunderbird 1.0.2 (Windows/20050317) 8, 22 -- X-Accept-Language: en-us, en 8, 22 -- MIME-Version: 1.0 8, 22 -- To: microscopy-at-microscopy.com 8, 22 -- Subject: Stray X-rays in FE SEM? 8, 22 -- References: {200507122213.j6CMDHwb015857-at-ns.microscopy.com} 8, 22 -- In-Reply-To: {200507122213.j6CMDHwb015857-at-ns.microscopy.com} 8, 22 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 8, 22 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
==============================Original Headers============================== 19, 34 -- From eprincipe01-at-hotmail.com Tue Jul 12 23:22:57 2005 19, 34 -- Received: from hotmail.com (bay0-dav-040.bay0.hotmail.com [64.4.60.62]) 19, 34 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6D4Mvqm006821 19, 34 -- for {microscopy-at-microscopy.com} ; Tue, 12 Jul 2005 23:22:57 -0500 19, 34 -- Received: from mail pickup service by hotmail.com with Microsoft SMTPSVC; 19, 34 -- Tue, 12 Jul 2005 21:22:54 -0700 19, 34 -- Message-ID: {BAY0-DAV-0403BFF608A882A9ABCE754B1DE0-at-phx.gbl} 19, 34 -- Received: from 207.68.174.36 by BAY0-DAV-040.phx.gbl with DAV; 19, 34 -- Wed, 13 Jul 2005 04:22:53 +0000 19, 34 -- X-Originating-IP: [207.68.174.36] 19, 34 -- X-Originating-Email: [eprincipe01-at-hotmail.com] 19, 34 -- X-Sender: eprincipe01-at-hotmail.com 19, 34 -- From: "Edward Principe" {eprincipe01-at-hotmail.com} 19, 34 -- Cc: 19, 34 -- Date: Tue, 12 Jul 2005 21:22:54 -0700 19, 34 -- thread-index: AcWHYbjkNc3aUdtgRZWNu5tAwUFXVg== 19, 34 -- To: {microscopy-at-microscopy.com} 19, 34 -- Thread-Topic: [Microscopy] Stray X-rays in FE SEM? 19, 34 -- MIME-Version: 1.0 19, 34 -- Content-Type: text/plain; 19, 34 -- charset="iso-8859-1" 19, 34 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1106 19, 34 -- X-Message-Status: n 19, 34 -- X-SID-PRA: hanke-at-mee-inc.com 19, 34 -- X-SID-Result: TempError 19, 34 -- X-Message-Info: JGTYoYF78jGeUUdCkFIoKJh+2+AQwGIRJvW2yuJm+a0= 19, 34 -- Received: from ns.microscopy.com ([206.69.208.10]) by mc7-f18.hotmail.com with Microsoft SMTPSVC(6.0.3790.211); Tue, 12 Jul 2005 16:03:33 -0700,from ns.microscopy.com (localhost.localdomain [127.0.0.1]) by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6CN3XLw032069 for {eprincipe01-at-hotmail.com} ; Tue, 12 Jul 2005 18:03:33 -0500,(from mail-at-localhost) by ns.microscopy.com (8.12.11/8.12.10/Submit) id j6CN3Xks032067; Tue, 12 Jul 2005 18:03:33 -0500 19, 34 -- X-Resent-From: "Microscopy Listserver" {microscopy-at-microscopy.com} 19, 34 -- Subject: RE: [Microscopy] Stray X-rays in FE SEM? 19, 34 -- Errors-to: MicroscopyListSpamFilter-at-microscopy.com 19, 34 -- X-lewp: MicroscopyListSpam NAGS 19, 34 -- X-OriginalArrivalTime: 12 Jul 2005 23:03:33.0917 (UTC) FILETIME=[ED98FCD0:01C58735] 19, 34 -- Content-Transfer-Encoding: 8bit 19, 34 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id j6D4Mvqm006821 ==============================End of - Headers==============================
I had a bad experience with formvar-copper grids in immunogold experiment many years ago when incubating primary antibody overnight and the rest done on the following morning. I am using Nickel grids now -- no more problems.
Ann Fook Yang EM Unit/ Unite EM Agriculture and Agri-Food Canada/Agriculture et Agroalimentaire Canada Telephone/Téléphone: 613-759-1638 Facsimile/Télécopieur: 613-759-1701 960 Carling Ave/960 Boul Carling Ottawa,Ontario/Ottawa, Ontario Canada K1A 0C6 yanga-at-agr.gc.ca
Agriculture and Agri-Food Canada - Agriculture et Agroalimentaire Canada
-----Original Message----- X-from: Rosemary.White-at-csiro.au [mailto:Rosemary.White-at-csiro.au] Sent: Tuesday, July 12, 2005 6:55 PM To: Yang, Ann-Fook
Dear Randy,
I only ran into the "use only Ni or Au grids" rule here, in my current job, and my predecessor certainly produced some superb images showing gold labelling of TEM sections. Blissfully unaware of this rule, I had been using copper grids for all EM immunolabelling. To get good labelling of one particular structure, which is about 40 nm diameter, I used uncoated thin-bar Cu grids so I could get labelling on both sides of the section - seemed to work just fine. I'll be interested to see the responses to your question.
cheers, Rosemary
Dr. Rosemary White rosemary.white-at-csiro.au Microscopy Centre ph. 61-2-6246 5475 CSIRO Plant Industry mob. 61-0402 835 973 GPO Box 1600 fax. 61-2-6246 5334 Canberra, ACT 2601 Australia
} From: TindallR-at-missouri.edu } Reply-To: microscopy-at-microscopy.com } Date: Tue, 12 Jul 2005 17:15:38 -0500 } To: rosemary.white-at-csiro.au } Subject: [Microscopy] TEM: Immunocytochemistry and Cu grids } } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Dear Listers, } } I have another one of my "back to the basics" questions that probably } make people wonder how the devil I ever got into this business in the } first place, but here goes anyway. } } We have a client who prepares his own blocks and brings them to us for } sectioning, staining, viewing, etc. One day he came into the lab with a } bunch of sections on copper mesh grids which he had done } immunocytochemistry with standard gold-conjugated secondaries. After } viewing them, I offered him the standard advice that next time we would } mount some sections on nickel or gold grids if he let us know in advance } that he would be doing immunocytochemistry. He said he always used } copper and asked why he should switch. My answer was something like } "Umm-uhhh.....because it's always done that way" with an additional } mumble about copper interfering with the labeling reaction somehow. } } Now I have another client, also infinitely more savvy in chemistry that } I am, asking the same question. } } So I checked through our little in-house research library (again) and } did some targeted Googling (again) and found that some folks say that } copper: 1) reacts with Tris-HCL buffer (okay, but we hardly ever use } that anyway); 2) may react with PBS buffers to produce fine } precipitates (but, but....doesn't that mean we should NEVER use Cu with } PBS?); 3) reacts with gold colloid (no other explanation given), 4) } can alter the charge distribution of the section and cause non-specific } background label; and 5) may be oxidized during labelling or interfere } with oxidation of chemical groups in the tissue to be labelled. Mostly } people just say to use Ni or Au without stating any reasons. } } Also, a glance through the literature shows that it's not uncommon to } find perfectly successful immunolabelling done on copper grids. } } Now, if I were determined to invade a country called Copper any or all } of the above reasons could be used with little further explanation, but } I'd like to know the Truth and pass it along to my admiring customers. } } Any takers? } } Thanks in advance. } } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility---We Do Small Well! } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-2227 } Email: tindallr-at-missouri.edu } Web: http://www.emc.missouri.edu } } } } } } } } ==============================Original Headers============================== } 15, 23 -- From TindallR-at-missouri.edu Tue Jul 12 17:12:03 2005 } 15, 23 -- Received: from um-exproto9.um.umsystem.edu } (um-exproto9.um.umsystem.edu [207.160.151.49]) } 15, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6CMC2XX014275 } 15, 23 -- for {microscopy-at-microscopy.com} ; Tue, 12 Jul 2005 17:12:03 -0500 } 15, 23 -- Received: from UM-EMAIL09.um.umsystem.edu ([207.160.151.31]) by } um-exproto9.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); } 15, 23 -- Tue, 12 Jul 2005 17:11:56 -0500 } 15, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 } 15, 23 -- Content-class: urn:content-classes:message } 15, 23 -- MIME-Version: 1.0 } 15, 23 -- Content-Type: text/plain; } 15, 23 -- charset="us-ascii" } 15, 23 -- Subject: TEM: Immunocytochemistry and Cu grids } 15, 23 -- Date: Tue, 12 Jul 2005 17:11:47 -0500 } 15, 23 -- Message-ID: } {BA876152E8653240BE8572E897083EE79802F3-at-UM-EMAIL09.um.umsystem.edu} } 15, 23 -- X-MS-Has-Attach: } 15, 23 -- X-MS-TNEF-Correlator: } 15, 23 -- Thread-Topic: TEM: Immunocytochemistry and Cu grids } 15, 23 -- Thread-Index: AcWHLrKQfT52UG1vRkSz6rcfHcA8yA== } 15, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} } 15, 23 -- To: {microscopy-at-microscopy.com} } 15, 23 -- X-OriginalArrivalTime: 12 Jul 2005 22:11:56.0360 (UTC) } FILETIME=[B74F4C80:01C5872E] } 15, 23 -- Content-Transfer-Encoding: 8bit } 15, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } ns.microscopy.com id j6CMC2XX014275 } ==============================End of - Headers============================== }
==============================Original Headers============================== 6, 23 -- From Rosemary.White-at-csiro.au Tue Jul 12 17:53:34 2005 6, 23 -- Received: from vic-ironport-ext-out2.csiro.au (vic-ironport-ext-out2.csiro.au [150.229.64.38]) 6, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6CMrXDw022373 6, 23 -- for {microscopy-at-microscopy.com} ; Tue, 12 Jul 2005 17:53:34 -0500 6, 23 -- Received: from exgw1-cbr.nexus.csiro.au (152.83.3.66) 6, 23 -- by vic-ironport-ext-out2.csiro.au with ESMTP; 13 Jul 2005 08:53:32 +1000 6, 23 -- X-BrightmailFiltered: true 6, 23 -- X-Brightmail-Tracker: AAAAAQAAA+k= 6, 23 -- X-IronPort-AV: i="3.93,284,1114956000"; 6, 23 -- d="scan'208"; a="45339190:sNHT23254752" 6, 23 -- Received: from [152.83.167.45] ([152.83.167.45]) by exgw1-cbr.nexus.csiro.au with Microsoft SMTPSVC(5.0.2195.6713); 6, 23 -- Wed, 13 Jul 2005 08:53:31 +1000 6, 23 -- User-Agent: Microsoft-Entourage/10.1.4.030702.0 6, 23 -- Date: Wed, 13 Jul 2005 08:54:42 +1000 6, 23 -- Subject: Re: [Microscopy] TEM: Immunocytochemistry and Cu grids 6, 23 -- From: Rosemary White {Rosemary.White-at-csiro.au} 6, 23 -- To: {microscopy-at-microscopy.com} 6, 23 -- Message-ID: {BEFA8752.108B7%Rosemary.White-at-csiro.au} 6, 23 -- In-Reply-To: {200507122215.j6CMFcFZ019486-at-ns.microscopy.com} 6, 23 -- Mime-version: 1.0 6, 23 -- Content-type: text/plain; charset="US-ASCII" 6, 23 -- Content-transfer-encoding: 7bit 6, 23 -- X-OriginalArrivalTime: 12 Jul 2005 22:53:31.0241 (UTC) FILETIME=[865FED90:01C58734] ==============================End of - Headers==============================
==============================Original Headers============================== 14, 29 -- From YANGA-at-AGR.GC.CA Wed Jul 13 08:24:43 2005 14, 29 -- Received: from agroutsmtp1.agr.gc.ca (agroutsmtp1.agr.gc.ca [192.197.71.175]) 14, 29 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6DDOhbQ020584 14, 29 -- for {microscopy-at-microscopy.com} ; Wed, 13 Jul 2005 08:24:43 -0500 14, 29 -- Received: from agrin1-old.agr.gc.ca (agrgate.agr.ca [192.197.71.189]) 14, 29 -- by agroutsmtp1.agr.gc.ca (8.12.8/8.12.8) with ESMTP id j6DDOg8Y024546 14, 29 -- for {microscopy-at-microscopy.com} ; Wed, 13 Jul 2005 09:24:42 -0400 14, 29 -- Received: from onncrxcn1.AGR.GC.CA ([10.117.15.130]) 14, 29 -- by agrin1-old.agr.gc.ca (8.12.8/8.12.8) with ESMTP id j6DDOcOO023305 14, 29 -- for {microscopy-at-microscopy.com} ; Wed, 13 Jul 2005 09:24:38 -0400 14, 29 -- Received: from onncrxms3.AGR.GC.CA ([10.117.15.30]) by onncrxcn1.AGR.GC.CA with Microsoft SMTPSVC(5.0.2195.6713); 14, 29 -- Wed, 13 Jul 2005 09:25:17 -0400 14, 29 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 14, 29 -- content-class: urn:content-classes:message 14, 29 -- MIME-Version: 1.0 14, 29 -- Content-Type: text/plain; 14, 29 -- charset="iso-8859-1" 14, 29 -- Subject: RE: [Microscopy] Re: TEM: Immunocytochemistry and Cu grids 14, 29 -- Date: Wed, 13 Jul 2005 09:25:16 -0400 14, 29 -- Message-ID: {E035A9C87303AE4AB9BA10FD8324DFB13553AA-at-onncrxms3.agr.gc.ca} 14, 29 -- X-MS-Has-Attach: 14, 29 -- X-MS-TNEF-Correlator: 14, 29 -- Thread-Topic: [Microscopy] Re: TEM: Immunocytochemistry and Cu grids 14, 29 -- Thread-Index: AcWHNM6wUJRJ1yarRAuufTkDb80FgAAdzEcg 14, 29 -- From: "Yang, Ann-Fook" {YANGA-at-AGR.GC.CA} 14, 29 -- To: {microscopy-at-microscopy.com} 14, 29 -- X-OriginalArrivalTime: 13 Jul 2005 13:25:17.0171 (UTC) FILETIME=[4F237430:01C587AE] 14, 29 -- Content-Transfer-Encoding: 8bit 14, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id j6DDOhbQ020584 ==============================End of - Headers==============================
We have had problems with Cu grids, even Formvar coated ones. I have precoated copper grids with Parlodion by dipping, blotting and drying before use, with or with out a formvar coating. THat method seems to protect the copper from reacting with PBS or whatever. This is not original to me, but I cannot recall where I read this (among many other things I can no longer recall). I think it might have been one of those smart Swiss guys.
Greg
TindallR-at-missouri.edu wrote:
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 6, 23 -- From gwe-at-ufl.edu Wed Jul 13 08:39:16 2005 6, 23 -- Received: from smtp.ufl.edu (sp42en1.nerdc.ufl.edu [128.227.74.42]) 6, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6DDdFCv028580 6, 23 -- for {microscopy-at-microscopy.com} ; Wed, 13 Jul 2005 08:39:15 -0500 6, 23 -- Received: from [127.0.0.1] (pc2524c.dhcp.clas.ufl.edu [128.227.60.197]) 6, 23 -- by smtp.ufl.edu (8.13.1/8.13.1/2.5.0) with ESMTP id j6DDdCog123902 6, 23 -- for {microscopy-at-microscopy.com} ; Wed, 13 Jul 2005 09:39:14 -0400 6, 23 -- Message-ID: {42D51989.90109-at-ufl.edu} 6, 23 -- Date: Wed, 13 Jul 2005 09:39:21 -0400 6, 23 -- From: Greg Erdos {gwe-at-ufl.edu} 6, 23 -- User-Agent: Mozilla Thunderbird 1.0.2 (Windows/20050317) 6, 23 -- X-Accept-Language: en-us, en 6, 23 -- MIME-Version: 1.0 6, 23 -- To: microscopy-at-microscopy.com 6, 23 -- Subject: Re: [Microscopy] TEM: Immunocytochemistry and Cu grids 6, 23 -- References: {200507122213.j6CMD58B015666-at-ns.microscopy.com} 6, 23 -- In-Reply-To: {200507122213.j6CMD58B015666-at-ns.microscopy.com} 6, 23 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 6, 23 -- Content-Transfer-Encoding: 7bit 6, 23 -- X-Spam-Status: hits=-0.904, required=5, tests=BAYES_30 6, 23 -- X-UFL-Spam-Status: hits=-0.904, required=5, tests=BAYES_30 6, 23 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) 6, 23 -- X-UFL-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) ==============================End of - Headers==============================
What's the big deal about preferring Cu to Ni anyway? Right, there is the charging problem with Nickel, but if you use anti-magnetic tweezers they are just as easy to handle than copper. And the price is really not that much more! We sometimes have to leave grids overnight or longer on solutions, so to be safe we use nickel for this. Since we don't want to make and keep separate stocks of copper and nickel, we switched entirely to nickel grids, and have had no problems with any of our applications. Am I missing something here?!!
Marc
On Tuesday, July 12, 2005, at 06:13 PM, TindallR-at-missouri.edu wrote:
} } } } ----------------------------------------------------------------------- } ----- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } ----- } } Dear Listers, } } I have another one of my "back to the basics" questions that probably } make people wonder how the devil I ever got into this business in the } first place, but here goes anyway. } } We have a client who prepares his own blocks and brings them to us for } sectioning, staining, viewing, etc. One day he came into the lab with } a } bunch of sections on copper mesh grids which he had done } immunocytochemistry with standard gold-conjugated secondaries. After } viewing them, I offered him the standard advice that next time we would } mount some sections on nickel or gold grids if he let us know in } advance } that he would be doing immunocytochemistry. He said he always used } copper and asked why he should switch. My answer was something like } "Umm-uhhh.....because it's always done that way" with an additional } mumble about copper interfering with the labeling reaction somehow. } } Now I have another client, also infinitely more savvy in chemistry that } I am, asking the same question. } } So I checked through our little in-house research library (again) and } did some targeted Googling (again) and found that some folks say that } copper: 1) reacts with Tris-HCL buffer (okay, but we hardly ever use } that anyway); 2) may react with PBS buffers to produce fine } precipitates (but, but....doesn't that mean we should NEVER use Cu with } PBS?); 3) reacts with gold colloid (no other explanation given), 4) } can alter the charge distribution of the section and cause non-specific } background label; and 5) may be oxidized during labelling or } interfere } with oxidation of chemical groups in the tissue to be labelled. } Mostly } people just say to use Ni or Au without stating any reasons. } } Also, a glance through the literature shows that it's not uncommon to } find perfectly successful immunolabelling done on copper grids. } } Now, if I were determined to invade a country called Copper any or all } of the above reasons could be used with little further explanation, but } I'd like to know the Truth and pass it along to my admiring customers. } } Any takers? } } Thanks in advance. } } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility---We Do Small Well! } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-2227 } Email: tindallr-at-missouri.edu } Web: http://www.emc.missouri.edu } } } } } } } } ==============================Original } Headers============================== } 15, 23 -- From TindallR-at-missouri.edu Tue Jul 12 17:12:03 2005 } 15, 23 -- Received: from um-exproto9.um.umsystem.edu } (um-exproto9.um.umsystem.edu [207.160.151.49]) } 15, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id } j6CMC2XX014275 } 15, 23 -- for {microscopy-at-microscopy.com} ; Tue, 12 Jul 2005 17:12:03 } -0500 } 15, 23 -- Received: from UM-EMAIL09.um.umsystem.edu ([207.160.151.31]) } by um-exproto9.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); } 15, 23 -- Tue, 12 Jul 2005 17:11:56 -0500 } 15, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 } 15, 23 -- Content-class: urn:content-classes:message } 15, 23 -- MIME-Version: 1.0 } 15, 23 -- Content-Type: text/plain; } 15, 23 -- charset="us-ascii" } 15, 23 -- Subject: TEM: Immunocytochemistry and Cu grids } 15, 23 -- Date: Tue, 12 Jul 2005 17:11:47 -0500 } 15, 23 -- Message-ID: } {BA876152E8653240BE8572E897083EE79802F3-at-UM-EMAIL09.um.umsystem.edu} } 15, 23 -- X-MS-Has-Attach: } 15, 23 -- X-MS-TNEF-Correlator: } 15, 23 -- Thread-Topic: TEM: Immunocytochemistry and Cu grids } 15, 23 -- Thread-Index: AcWHLrKQfT52UG1vRkSz6rcfHcA8yA== } 15, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} } 15, 23 -- To: {microscopy-at-microscopy.com} } 15, 23 -- X-OriginalArrivalTime: 12 Jul 2005 22:11:56.0360 (UTC) } FILETIME=[B74F4C80:01C5872E] } 15, 23 -- Content-Transfer-Encoding: 8bit } 15, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } ns.microscopy.com id j6CMC2XX014275 } ==============================End of - } Headers============================== } }
-- 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
==============================Original Headers============================== 6, 18 -- From marc.pypaert-at-yale.edu Wed Jul 13 10:37:11 2005 6, 18 -- Received: from BIOMED.MED.YALE.EDU (biomed.med.yale.edu [130.132.232.48]) 6, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6DFbBff005748 6, 18 -- for {microscopy-at-microscopy.com} ; Wed, 13 Jul 2005 10:37:11 -0500 6, 18 -- Received: from yale.edu (net234-111.med.yale.edu [130.132.234.111]) 6, 18 -- by biomed.med.yale.edu (PMDF V6.1-1 #30532) 6, 18 -- with ESMTP id {01LQKPAV9P3Y007BQ2-at-biomed.med.yale.edu} for 6, 18 -- microscopy-at-microscopy.com; Wed, 13 Jul 2005 11:20:03 -0400 (EDT) 6, 18 -- Date: Wed, 13 Jul 2005 11:19:42 -0400 6, 18 -- From: Marc Pypaert {marc.pypaert-at-yale.edu} 6, 18 -- Subject: Re: [Microscopy] TEM: Immunocytochemistry and Cu grids 6, 18 -- In-reply-to: {200507122213.j6CMDcxr016073-at-ns.microscopy.com} 6, 18 -- To: microscopy-at-microscopy.com 6, 18 -- Message-id: {896F9E09-F3B1-11D9-AC3D-0030659833B4-at-yale.edu} 6, 18 -- MIME-version: 1.0 (Apple Message framework v553) 6, 18 -- X-Mailer: Apple Mail (2.553) 6, 18 -- Content-type: text/plain; format=flowed; charset=US-ASCII; delsp=yes 6, 18 -- Content-transfer-encoding: 7bit ==============================End of - Headers==============================
It doesn't make a lot of difference to me either way, except to be able to actually have an intelligent explanation when our users ask questions about our methods. That said, there are times when we do get severe image distortion in our TEM when using Ni grids with a healthy charge on them. But my main motivation was curiosity, nada mas.
Randy
-----Original Message----- X-from: marc.pypaert-at-yale.edu [mailto:marc.pypaert-at-yale.edu] Sent: Wednesday, July 13, 2005 10:39 AM To: Tindall, Randy D.
What's the big deal about preferring Cu to Ni anyway? Right, there is the charging problem with Nickel, but if you use anti-magnetic tweezers they are just as easy to handle than copper. And the price is really not that much more! We sometimes have to leave grids overnight or longer on solutions, so to be safe we use nickel for this. Since we don't want to make and keep separate stocks of copper and nickel, we switched entirely to nickel grids, and have had no problems with any of our applications. Am I missing something here?!!
Marc
On Tuesday, July 12, 2005, at 06:13 PM, TindallR-at-missouri.edu wrote:
} } } } ---------------------------------------------------------------------- } - } ----- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } - } ----- } } Dear Listers, } } I have another one of my "back to the basics" questions that probably } make people wonder how the devil I ever got into this business in the } first place, but here goes anyway. } } We have a client who prepares his own blocks and brings them to us for
} sectioning, staining, viewing, etc. One day he came into the lab with
} a bunch of sections on copper mesh grids which he had done } immunocytochemistry with standard gold-conjugated secondaries. After } viewing them, I offered him the standard advice that next time we } would mount some sections on nickel or gold grids if he let us know in
} advance that he would be doing immunocytochemistry. He said he always
} used copper and asked why he should switch. My answer was something } like "Umm-uhhh.....because it's always done that way" with an } additional mumble about copper interfering with the labeling reaction } somehow. } } Now I have another client, also infinitely more savvy in chemistry } that I am, asking the same question. } } So I checked through our little in-house research library (again) and } did some targeted Googling (again) and found that some folks say that } copper: 1) reacts with Tris-HCL buffer (okay, but we hardly ever use } that anyway); 2) may react with PBS buffers to produce fine } precipitates (but, but....doesn't that mean we should NEVER use Cu with } PBS?); 3) reacts with gold colloid (no other explanation given), 4) } can alter the charge distribution of the section and cause } non-specific background label; and 5) may be oxidized during } labelling or interfere } with oxidation of chemical groups in the tissue to be labelled. } Mostly } people just say to use Ni or Au without stating any reasons. } } Also, a glance through the literature shows that it's not uncommon to } find perfectly successful immunolabelling done on copper grids. } } Now, if I were determined to invade a country called Copper any or all
} of the above reasons could be used with little further explanation, } but I'd like to know the Truth and pass it along to my admiring customers. } } Any takers? } } Thanks in advance. } } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility---We Do Small Well! } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-2227 } Email: tindallr-at-missouri.edu } Web: http://www.emc.missouri.edu } } } } } } } } ==============================Original } Headers============================== } 15, 23 -- From TindallR-at-missouri.edu Tue Jul 12 17:12:03 2005 15, 23 } -- Received: from um-exproto9.um.umsystem.edu } (um-exproto9.um.umsystem.edu [207.160.151.49]) } 15, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id } j6CMC2XX014275 } 15, 23 -- for {microscopy-at-microscopy.com} ; Tue, 12 Jul 2005 17:12:03 } -0500 } 15, 23 -- Received: from UM-EMAIL09.um.umsystem.edu ([207.160.151.31])
} by um-exproto9.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); } 15, 23 -- Tue, 12 Jul 2005 17:11:56 -0500 } 15, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 15, 23
-- 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
==============================Original Headers============================== 6, 18 -- From marc.pypaert-at-yale.edu Wed Jul 13 10:37:11 2005 6, 18 -- Received: from BIOMED.MED.YALE.EDU (biomed.med.yale.edu [130.132.232.48]) 6, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6DFbBff005748 6, 18 -- for {microscopy-at-microscopy.com} ; Wed, 13 Jul 2005 10:37:11 -0500 6, 18 -- Received: from yale.edu (net234-111.med.yale.edu [130.132.234.111]) 6, 18 -- by biomed.med.yale.edu (PMDF V6.1-1 #30532) 6, 18 -- with ESMTP id {01LQKPAV9P3Y007BQ2-at-biomed.med.yale.edu} for 6, 18 -- microscopy-at-microscopy.com; Wed, 13 Jul 2005 11:20:03 -0400 (EDT) 6, 18 -- Date: Wed, 13 Jul 2005 11:19:42 -0400 6, 18 -- From: Marc Pypaert {marc.pypaert-at-yale.edu} 6, 18 -- Subject: Re: [Microscopy] TEM: Immunocytochemistry and Cu grids 6, 18 -- In-reply-to: {200507122213.j6CMDcxr016073-at-ns.microscopy.com} 6, 18 -- To: microscopy-at-microscopy.com 6, 18 -- Message-id: {896F9E09-F3B1-11D9-AC3D-0030659833B4-at-yale.edu} 6, 18 -- MIME-version: 1.0 (Apple Message framework v553) 6, 18 -- X-Mailer: Apple Mail (2.553) 6, 18 -- Content-type: text/plain; format=flowed; charset=US-ASCII; delsp=yes 6, 18 -- Content-transfer-encoding: 7bit ==============================End of - Headers==============================
==============================Original Headers============================== 22, 23 -- From TindallR-at-missouri.edu Wed Jul 13 10:56:21 2005 22, 23 -- Received: from um-exproto8.um.umsystem.edu (um-exproto8.um.umsystem.edu [207.160.151.48]) 22, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6DFuLiE013730 22, 23 -- for {microscopy-at-microscopy.com} ; Wed, 13 Jul 2005 10:56:21 -0500 22, 23 -- Received: from UM-EMAIL09.um.umsystem.edu ([207.160.151.31]) by um-exproto8.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 22, 23 -- Wed, 13 Jul 2005 10:56:21 -0500 22, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 22, 23 -- Content-class: urn:content-classes:message 22, 23 -- MIME-Version: 1.0 22, 23 -- Content-Type: text/plain; 22, 23 -- charset="us-ascii" 22, 23 -- Subject: RE: [Microscopy] Re: TEM: Immunocytochemistry and Cu grids 22, 23 -- Date: Wed, 13 Jul 2005 10:56:20 -0500 22, 23 -- Message-ID: {BA876152E8653240BE8572E897083EE79802F4-at-UM-EMAIL09.um.umsystem.edu} 22, 23 -- X-MS-Has-Attach: 22, 23 -- X-MS-TNEF-Correlator: 22, 23 -- Thread-Topic: [Microscopy] Re: TEM: Immunocytochemistry and Cu grids 22, 23 -- Thread-Index: AcWHwPNRIL3xOIhMTey96wt+T7YehAAAdtIg 22, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 22, 23 -- To: {microscopy-at-microscopy.com} 22, 23 -- X-OriginalArrivalTime: 13 Jul 2005 15:56:21.0191 (UTC) FILETIME=[69B72570:01C587C3] 22, 23 -- Content-Transfer-Encoding: 8bit 22, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id j6DFuLiE013730 ==============================End of - Headers==============================
The main problem encountered with copper grids used for immunocytochemistry or immunogold labeling is the reaction of the copper with salts in the buffers used during labeling. This is a time-related reaction, and can usually be avoided by having short labeling runs and using grids with films on them. Gilder grids, available from major microscope supply vendors, are gold-coated copper grids, and are not that much more expensive than regular copper grids. These grids may be the variety used by researchers who have reported having no problems with their grids during their immuno runs. I usually use nickel grids for my work, but have used formvar-coated copper grids without problems for 1 day immuno runs so long as the film is intact and as long as I don't get clumsy and end up sinking my grids in my solutions (if I do sink them, I do a quick distilled water rinse, blot the back of the grids with filter paper until almost dry, then re-float them where I left off). As others have shared, the nickel grids are much sturdier, not too expensive, and are not a problem to handle as long as you use antimagnetic forceps. With the nickel grids, remember to correct for astigmatism in the TEM by using a hole in your sample on the nickel grid. This will accommodate for any inherent residual magnetic field in the grid itself.
Edward Haller Lab Manager, Diagnostic Electron Microscopy Lab University of South Florida Pathology Department Tampa, FL 33612 (813) 974-9584
==============================Original Headers============================== 4, 26 -- From ehaller-at-hsc.usf.edu Wed Jul 13 11:22:28 2005 4, 26 -- Received: from hscantivirus.hsc.usf.edu (hscantivirus.hsc.usf.edu [131.247.67.157]) 4, 26 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id j6DGMRdH021865 4, 26 -- for {microscopy-at-microscopy.com} ; Wed, 13 Jul 2005 11:22:28 -0500 4, 26 -- Received: from HSCMAIL.hscnet.hsc.usf.edu ([IP=131.247.67.144]) by eSafe SMTP Relay 1121174809; Wed Jul 13 12:21:14 2005 4, 26 -- Received: from CONEXCHANGE.hscnet.hsc.usf.edu ([131.247.67.142]) by HSCMAIL.hscnet.hsc.usf.edu with Microsoft SMTPSVC(6.0.3790.0); 4, 26 -- Wed, 13 Jul 2005 12:19:37 -0400 4, 26 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 4, 26 -- Content-class: urn:content-classes:message 4, 26 -- MIME-Version: 1.0 4, 26 -- Content-Type: text/plain; 4, 26 -- charset="US-ASCII" 4, 26 -- Subject: RE: [Microscopy] TEM: Immunocytochemistry and Cu grids 4, 26 -- Date: Wed, 13 Jul 2005 12:19:37 -0400 4, 26 -- Message-ID: {841D767DCDE87C49A02DA6DFC0BABABF691823-at-COMEXCHANGE.hscnet.hsc.usf.edu} 4, 26 -- X-MS-Has-Attach: 4, 26 -- X-MS-TNEF-Correlator: 4, 26 -- Thread-Topic: [Microscopy] TEM: Immunocytochemistry and Cu grids 4, 26 -- Thread-Index: AcWHw4VKXyuYUb6ZSuiO7MbvaO3xNAAALgXg 4, 26 -- From: "Haller, Ed" {ehaller-at-hsc.usf.edu} 4, 26 -- To: {microscopy-at-microscopy.com} 4, 26 -- X-OriginalArrivalTime: 13 Jul 2005 16:19:37.0495 (UTC) FILETIME=[A9FA3A70:01C587C6] 4, 26 -- X-ESAFE-STATUS: Mail clean 4, 26 -- X-ESAFE-DETAILS: Clean 4, 26 -- Content-Transfer-Encoding: 8bit 4, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id j6DGMRdH021865 ==============================End of - Headers==============================
Sorry for any misunderstanding - My response was not really directed at you, but at the EM community in general who for some reason seems to be biased towards copper. I have never understood this. I have been doing EM for nearly 20 years, exclusively using nickel grids, and would really like to find out if I was mistaken this entire time!! Or is it just another case of "dogma", like Paul put in nicely in his posting? Anyway, I will read with interest the responses you get to your posting! Best
Marc
On Wednesday, July 13, 2005, at 11:57 AM, TindallR-at-missouri.edu wrote:
} } } } ----------------------------------------------------------------------- } ----- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } ----- } } Marc, } } It doesn't make a lot of difference to me either way, except to be able } to actually have an intelligent explanation when our users ask } questions } about our methods. That said, there are times when we do get severe } image distortion in our TEM when using Ni grids with a healthy charge } on } them. But my main motivation was curiosity, nada mas. } } Randy } } -----Original Message----- } X-from: marc.pypaert-at-yale.edu [mailto:marc.pypaert-at-yale.edu] } Sent: Wednesday, July 13, 2005 10:39 AM } To: Tindall, Randy D. } Subject: [Microscopy] Re: TEM: Immunocytochemistry and Cu grids } } } } } ----------------------------------------------------------------------- } - } ---- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } - } ---- } } What's the big deal about preferring Cu to Ni anyway? } Right, there is the charging problem with Nickel, but if you use } anti-magnetic tweezers they are just as easy to handle than copper. And } the price is really not that much more! We sometimes have to leave } grids } overnight or longer on solutions, so to be safe we use nickel for this. } Since we don't want to make and keep separate stocks of copper and } nickel, we switched entirely to nickel grids, and have had no problems } with any of our applications. Am I missing something here?!! } } Marc } } On Tuesday, July 12, 2005, at 06:13 PM, TindallR-at-missouri.edu wrote: } } } } } } } } } ---------------------------------------------------------------------- } } - } } ----- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } } America To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------- } } - } } ----- } } } } Dear Listers, } } } } I have another one of my "back to the basics" questions that probably } } make people wonder how the devil I ever got into this business in the } } first place, but here goes anyway. } } } } We have a client who prepares his own blocks and brings them to us for } } } sectioning, staining, viewing, etc. One day he came into the lab with } } } a bunch of sections on copper mesh grids which he had done } } immunocytochemistry with standard gold-conjugated secondaries. After } } viewing them, I offered him the standard advice that next time we } } would mount some sections on nickel or gold grids if he let us know in } } } advance that he would be doing immunocytochemistry. He said he always } } } used copper and asked why he should switch. My answer was something } } like "Umm-uhhh.....because it's always done that way" with an } } additional mumble about copper interfering with the labeling reaction } } somehow. } } } } Now I have another client, also infinitely more savvy in chemistry } } that I am, asking the same question. } } } } So I checked through our little in-house research library (again) and } } did some targeted Googling (again) and found that some folks say that } } copper: 1) reacts with Tris-HCL buffer (okay, but we hardly ever use } } that anyway); 2) may react with PBS buffers to produce fine } } precipitates (but, but....doesn't that mean we should NEVER use Cu } with } } PBS?); 3) reacts with gold colloid (no other explanation given), 4) } } can alter the charge distribution of the section and cause } } non-specific background label; and 5) may be oxidized during } } labelling or interfere } } with oxidation of chemical groups in the tissue to be labelled. } } Mostly } } people just say to use Ni or Au without stating any reasons. } } } } Also, a glance through the literature shows that it's not uncommon to } } find perfectly successful immunolabelling done on copper grids. } } } } Now, if I were determined to invade a country called Copper any or all } } } of the above reasons could be used with little further explanation, } } but I'd like to know the Truth and pass it along to my admiring } customers. } } } } Any takers? } } } } Thanks in advance. } } } } Randy } } } } Randy Tindall } } EM Specialist } } Electron Microscopy Core Facility---We Do Small Well! } } W122 Veterinary Medicine } } University of Missouri } } Columbia, MO 65211 } } Tel: (573) 882-8304 } } Fax: (573) 884-2227 } } Email: tindallr-at-missouri.edu } } Web: http://www.emc.missouri.edu } } } } } } } } } } } } } } } } ==============================Original } } Headers============================== } } 15, 23 -- From TindallR-at-missouri.edu Tue Jul 12 17:12:03 2005 15, 23 } } -- Received: from um-exproto9.um.umsystem.edu } } (um-exproto9.um.umsystem.edu [207.160.151.49]) } } 15, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id } } j6CMC2XX014275 } } 15, 23 -- for {microscopy-at-microscopy.com} ; Tue, 12 Jul 2005 } 17:12:03 } } -0500 } } 15, 23 -- Received: from UM-EMAIL09.um.umsystem.edu ([207.160.151.31]) } } } by um-exproto9.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); } } 15, 23 -- Tue, 12 Jul 2005 17:11:56 -0500 } } 15, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 15, 23 } } } -- Content-class: urn:content-classes:message 15, 23 -- MIME-Version: } } 1.0 15, 23 -- Content-Type: text/plain; } } 15, 23 -- charset="us-ascii" } } 15, 23 -- Subject: TEM: Immunocytochemistry and Cu grids 15, 23 -- } } Date: Tue, 12 Jul 2005 17:11:47 -0500 15, 23 -- Message-ID: } } {BA876152E8653240BE8572E897083EE79802F3-at-UM-EMAIL09.um.umsystem.edu} } } 15, 23 -- X-MS-Has-Attach: } } 15, 23 -- X-MS-TNEF-Correlator: } } 15, 23 -- Thread-Topic: TEM: Immunocytochemistry and Cu grids 15, 23 } } -- Thread-Index: AcWHLrKQfT52UG1vRkSz6rcfHcA8yA== 15, 23 -- From: } } "Tindall, Randy D." {TindallR-at-missouri.edu} 15, 23 -- To: } } {microscopy-at-microscopy.com} 15, 23 -- X-OriginalArrivalTime: 12 Jul } } 2005 22:11:56.0360 (UTC) FILETIME=[B74F4C80:01C5872E] 15, 23 -- } } Content-Transfer-Encoding: 8bit 15, 23 -- X-MIME-Autoconverted: from } } quoted-printable to 8bit by ns.microscopy.com id j6CMC2XX014275 } } ==============================End of - } } Headers============================== } } } } } } -- } 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 } } } ==============================Original } Headers============================== } 6, 18 -- From marc.pypaert-at-yale.edu Wed Jul 13 10:37:11 2005 6, 18 -- } Received: from BIOMED.MED.YALE.EDU (biomed.med.yale.edu } [130.132.232.48]) } 6, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id } j6DFbBff005748 } 6, 18 -- for {microscopy-at-microscopy.com} ; Wed, 13 Jul 2005 } 10:37:11 -0500 } 6, 18 -- Received: from yale.edu (net234-111.med.yale.edu } [130.132.234.111]) 6, 18 -- by biomed.med.yale.edu (PMDF V6.1-1 } #30532) } 6, 18 -- with ESMTP id {01LQKPAV9P3Y007BQ2-at-biomed.med.yale.edu} for 6, } 18 -- microscopy-at-microscopy.com; Wed, 13 Jul 2005 11:20:03 -0400 (EDT) } 6, 18 -- Date: Wed, 13 Jul 2005 11:19:42 -0400 6, 18 -- From: Marc } Pypaert {marc.pypaert-at-yale.edu} 6, 18 -- Subject: Re: [Microscopy] TEM: } Immunocytochemistry and Cu grids 6, 18 -- In-reply-to: } {200507122213.j6CMDcxr016073-at-ns.microscopy.com} } 6, 18 -- To: microscopy-at-microscopy.com } 6, 18 -- Message-id: {896F9E09-F3B1-11D9-AC3D-0030659833B4-at-yale.edu} } 6, 18 -- MIME-version: 1.0 (Apple Message framework v553) 6, 18 -- } X-Mailer: Apple Mail (2.553) 6, 18 -- Content-type: text/plain; } format=flowed; charset=US-ASCII; delsp=yes 6, 18 -- } Content-transfer-encoding: 7bit ==============================End of - } Headers============================== } } } ==============================Original } Headers============================== } 22, 23 -- From TindallR-at-missouri.edu Wed Jul 13 10:56:21 2005 } 22, 23 -- Received: from um-exproto8.um.umsystem.edu } (um-exproto8.um.umsystem.edu [207.160.151.48]) } 22, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id } j6DFuLiE013730 } 22, 23 -- for {microscopy-at-microscopy.com} ; Wed, 13 Jul 2005 10:56:21 } -0500 } 22, 23 -- Received: from UM-EMAIL09.um.umsystem.edu ([207.160.151.31]) } by um-exproto8.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); } 22, 23 -- Wed, 13 Jul 2005 10:56:21 -0500 } 22, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 } 22, 23 -- Content-class: urn:content-classes:message } 22, 23 -- MIME-Version: 1.0 } 22, 23 -- Content-Type: text/plain; } 22, 23 -- charset="us-ascii" } 22, 23 -- Subject: RE: [Microscopy] Re: TEM: Immunocytochemistry and } Cu grids } 22, 23 -- Date: Wed, 13 Jul 2005 10:56:20 -0500 } 22, 23 -- Message-ID: } {BA876152E8653240BE8572E897083EE79802F4-at-UM-EMAIL09.um.umsystem.edu} } 22, 23 -- X-MS-Has-Attach: } 22, 23 -- X-MS-TNEF-Correlator: } 22, 23 -- Thread-Topic: [Microscopy] Re: TEM: Immunocytochemistry and } Cu grids } 22, 23 -- Thread-Index: AcWHwPNRIL3xOIhMTey96wt+T7YehAAAdtIg } 22, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} } 22, 23 -- To: {microscopy-at-microscopy.com} } 22, 23 -- X-OriginalArrivalTime: 13 Jul 2005 15:56:21.0191 (UTC) } FILETIME=[69B72570:01C587C3] } 22, 23 -- Content-Transfer-Encoding: 8bit } 22, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } ns.microscopy.com id j6DFuLiE013730 } ==============================End of - } Headers============================== } }
-- 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
==============================Original Headers============================== 7, 18 -- From marc.pypaert-at-yale.edu Wed Jul 13 12:01:02 2005 7, 18 -- Received: from BIOMED.MED.YALE.EDU (biomed.med.yale.edu [130.132.232.48]) 7, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6DH11b8030049 7, 18 -- for {microscopy-at-microscopy.com} ; Wed, 13 Jul 2005 12:01:02 -0500 7, 18 -- Received: from yale.edu (net234-111.med.yale.edu [130.132.234.111]) 7, 18 -- by biomed.med.yale.edu (PMDF V6.1-1 #30532) 7, 18 -- with ESMTP id {01LQKSJRTF2K009625-at-biomed.med.yale.edu} for 7, 18 -- microscopy-at-microscopy.com; Wed, 13 Jul 2005 12:52:45 -0400 (EDT) 7, 18 -- Date: Wed, 13 Jul 2005 12:52:22 -0400 7, 18 -- From: Marc Pypaert {marc.pypaert-at-yale.edu} 7, 18 -- Subject: Re: [Microscopy] TEM: Immunocytochemistry and Cu grids 7, 18 -- In-reply-to: {200507131557.j6DFv7KK015520-at-ns.microscopy.com} 7, 18 -- To: microscopy-at-microscopy.com 7, 18 -- Message-id: {7B66628D-F3BE-11D9-AC3D-0030659833B4-at-yale.edu} 7, 18 -- MIME-version: 1.0 (Apple Message framework v553) 7, 18 -- X-Mailer: Apple Mail (2.553) 7, 18 -- Content-type: text/plain; format=flowed; charset=US-ASCII; delsp=yes 7, 18 -- Content-transfer-encoding: 7bit ==============================End of - Headers==============================
Over the years I have frequently seen cases where the samples themselves have reacted with the copper grid. Two examples:
1) Looking at Fe-S precipitates in magnetotactic bacteria, when the precipitates were ugly and analyzed as copper sulphide. Our surmise was that the iron and copper had undergone ion exchange while the grid (and carbon film) was wet with the culture medium. When we used nickel grids, the precipitates were well formed and composed of iron and sulphur.
2) Looking at Si precipitates in Al-Si electronic bond wire. The specimens were prepared by embedding and microtoming, floating in DI water. The Si precipitates were surrounded by an ugly mess containing masses of copper. Again, presumed to me electrochemical reaction between the Al and the Cu grid, through the DI water medium, somehow preferentially at the Si precipitates. Again, use of Ni grids produced good pictures allowing us to characterize the precipitation.
Having said all that, we continue to default to use copper grids!
Tony.
*********************************************** Anthony J. Garratt-Reed, M.A., D.Phil. MIT Room 13-1027 77 Massachusetts Avenue Cambridge, MA 02139-4307 USA
Thanks for your weigh-in on this subject. I was sorry to learn of your hearing problems; has this been coming on gradually or did something happen (i.e. a severe cold, etc.)to bring the hearing loss on?
I was surprised to learn that Boston is not yet out of the running for an MSA meeting! If I can help in any way, I'll throw my hat in the ring.
Hope your summer (now that it is the middle of July!) is going well.
Peggy
Peggy Sherwood Lab Associate, Photopathology Wellman Center for Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
-----Original Message----- X-from: tonygr-at-MIT.EDU [mailto:tonygr-at-MIT.EDU] Sent: Wednesday, July 13, 2005 1:33 PM To: Sherwood, Margaret
Over the years I have frequently seen cases where the samples themselves have reacted with the copper grid. Two examples:
1) Looking at Fe-S precipitates in magnetotactic bacteria, when the precipitates were ugly and analyzed as copper sulphide. Our surmise was that the iron and copper had undergone ion exchange while the grid (and carbon film) was wet with the culture medium. When we used nickel grids, the precipitates were well formed and composed of iron and sulphur.
2) Looking at Si precipitates in Al-Si electronic bond wire. The specimens were prepared by embedding and microtoming, floating in DI water. The Si precipitates were surrounded by an ugly mess containing masses of copper. Again, presumed to me electrochemical reaction between the Al and the Cu grid, through the DI water medium, somehow preferentially at the Si precipitates. Again, use of Ni grids produced good pictures allowing us to characterize the precipitation.
Having said all that, we continue to default to use copper grids!
Tony.
*********************************************** Anthony J. Garratt-Reed, M.A., D.Phil. MIT Room 13-1027 77 Massachusetts Avenue Cambridge, MA 02139-4307 USA
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (tpepper-at-iastate.edu) from http://www.microscopy.org/MicroscopyListserver/MLFormMail.html on Wednesday, July 13, 2005 at 09:21:59 ---------------------------------------------------------------------------
Email: tpepper-at-iastate.edu Name: Tracey Pepper
Organization: Iowa State University
Title-Subject: [Filtered] MListserver:
Question: HELP! We are in desparate need of some assisitance! We have a scientist in Texas who is in the field collecting crucial samples for TEM examination. They must be collected in the next couple of days. Our efforts to send (via DHL) good fixative (standard em fix for plants a double aldehyde in a 0.1 M cacodylate) have failed! The person is located in Plainview, Texas. Is there anyone on this listserver who could make us some fixtive (150 mls)for this person to come pick up? Preferably within an hour radius? If you can, Please call 515-294-3872 as soon as possible. We would be in your debt!! With Great appreciation, Tracey Pepper Bessey Microscopy Facility Iowa State University Ames, IA 50011-1020
Your best bet would be to call some one at Texas Tech in Lubbock http://www.macmed.ttuhsc.edu/ They may be more than an hour from Plainview but that close to the edge of the world. Your only other chance is Amarillo with the TAM Vet Med Diagnostic lab http://www.medcenter.org/facilities/am-vetlab.html.
Good luck Gordon
Gordon Couger I collect links on information related to light microscopes. www.couger.com/microscope/links/gclinks.html Please forward anything you think might be useful to others. Microscope Documentation is at www.science-info.org
tpepper-at-iastate.edu wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by } (tpepper-at-iastate.edu) from http://www.microscopy.org/MicroscopyListserver/MLFormMail.html } on Wednesday, July 13, 2005 at 09:21:59 } --------------------------------------------------------------------------- } } Email: tpepper-at-iastate.edu } Name: Tracey Pepper } } Organization: Iowa State University } } Title-Subject: [Filtered] MListserver: } } Question: HELP! We are in desparate need of some assisitance! We have a scientist in Texas who is in the field collecting crucial samples for TEM examination. They must be collected in the next couple of days. Our efforts to send (via DHL) good fixative (standard em fix for plants a double aldehyde in a 0.1 M cacodylate) have failed! The person is located in Plainview, Texas. Is there anyone on this listserver who could make us some fixtive (150 mls)for this person to come pick up? Preferably within an hour radius? If you can, Please call 515-294-3872 as soon as possible. We would be in your debt!! } With Great appreciation, } Tracey Pepper } Bessey Microscopy Facility } Iowa State University } Ames, IA 50011-1020 } } --------------------------------------------------------------------------- } } ==============================Original Headers============================== } 6, 12 -- From zaluzec-at-microscopy.com Wed Jul 13 13:54:01 2005 } 6, 12 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) } 6, 12 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6DIs1ei022223 } 6, 12 -- for {microscopy-at-microscopy.com} ; Wed, 13 Jul 2005 13:54:01 -0500 } 6, 12 -- Mime-Version: 1.0 } 6, 12 -- X-Sender: (Unverified) } 6, 12 -- Message-Id: {p06110400befb1371ac5b-at-[206.69.208.22]} } 6, 12 -- Date: Wed, 13 Jul 2005 13:54:00 -0500 } 6, 12 -- To: microscopy-at-microscopy.com } 6, 12 -- From: tpepper-at-iastate.edu (by way of MicroscopyListserver) } 6, 12 -- Subject: viaWWW: Help with Fixative near Plainview Texas } 6, 12 -- Content-Type: text/plain; charset="us-ascii" } ==============================End of - Headers============================== } }
==============================Original Headers============================== 5, 20 -- From gcc-at-couger.com Wed Jul 13 15:01:27 2005 5, 20 -- Received: from lakermmtao08.cox.net (lakermmtao08.cox.net [68.230.240.31]) 5, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6DK1RJr030657 5, 20 -- for {microscopy-at-microscopy.com} ; Wed, 13 Jul 2005 15:01:27 -0500 5, 20 -- Received: from [127.0.0.1] (really [68.12.43.168]) by lakermmtao08.cox.net 5, 20 -- (InterMail vM.6.01.04.00 201-2131-118-20041027) with ESMTP 5, 20 -- id {20050713200117.YBBU19415.lakermmtao08.cox.net-at-[127.0.0.1]} 5, 20 -- for {microscopy-at-microscopy.com} ; Wed, 13 Jul 2005 16:01:17 -0400 5, 20 -- Message-ID: {42D5730D.4090609-at-couger.com} 5, 20 -- Date: Wed, 13 Jul 2005 15:01:17 -0500 5, 20 -- From: Gordon Couger {gcc-at-couger.com} 5, 20 -- User-Agent: Mozilla Thunderbird 1.0.2 (Windows/20050317) 5, 20 -- X-Accept-Language: en-us, en 5, 20 -- MIME-Version: 1.0 5, 20 -- To: microscopy-at-microscopy.com 5, 20 -- Subject: Re: [Microscopy] viaWWW: Help with Fixative near Plainview Texas 5, 20 -- References: {200507131858.j6DIw5Mo027248-at-ns.microscopy.com} 5, 20 -- In-Reply-To: {200507131858.j6DIw5Mo027248-at-ns.microscopy.com} 5, 20 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 5, 20 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Does anyone have a Nikon Diaphot epi-fluorescence lamp house and filter set/holder for sale? Alternatively, if anyone knows of a second hand microscope parts vendor?
Thanks for your help.
Regards
Andrew
________________________________________________________________________ ______ Andrew McNaughton Otago Centre for Confocal Microscopy C/-Department of Anatomy and Structural Biology School of Medical Sciences University of Otago PO Box 913, Dunedin NEW ZEALAND
I am using titanium tweezers with nickel grids - they are cheap nowadays and I like them (tweezers) for light weight and soft action. I am using carbon coating on top of all my plastic films (copper or nickel grids) and have no problems with charge/astigmatism. I suspect, charging problem happens when grids quite old and/or dirty. Sergey
At 09:23 AM 7/13/2005, you wrote:
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==============================Original Headers============================== 6, 25 -- From sryazant-at-ucla.edu Wed Jul 13 16:09:43 2005 6, 25 -- Received: from smtp-3.smtp.ucla.edu (smtp-3.smtp.ucla.edu [169.232.48.136]) 6, 25 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6DL9gde014359 6, 25 -- for {microscopy-at-microscopy.com} ; Wed, 13 Jul 2005 16:09:43 -0500 6, 25 -- Received: from mail.ucla.edu (mail.ucla.edu [169.232.48.141]) 6, 25 -- by smtp-3.smtp.ucla.edu (8.13.4/8.13.4) with ESMTP id j6DL9gFM012306 6, 25 -- for {microscopy-at-microscopy.com} ; Wed, 13 Jul 2005 14:09:42 -0700 6, 25 -- Received: from kopoba.ucla.edu (ts13-161.dialup.bol.ucla.edu [169.232.229.108]) 6, 25 -- (authenticated bits=0) 6, 25 -- by mail.ucla.edu (8.13.4/8.13.4) with ESMTP id j6DL9YYN006121 6, 25 -- (version=TLSv1/SSLv3 cipher=DES-CBC3-SHA bits=168 verify=NOT) 6, 25 -- for {microscopy-at-microscopy.com} ; Wed, 13 Jul 2005 14:09:38 -0700 6, 25 -- Message-Id: {6.1.2.0.2.20050713140435.032c38b0-at-mail.ucla.edu} 6, 25 -- X-Sender: sryazant-at-mail.ucla.edu 6, 25 -- X-Mailer: QUALCOMM Windows Eudora Version 6.1.2.0 6, 25 -- Date: Wed, 13 Jul 2005 14:09:25 -0700 6, 25 -- To: microscopy-at-microscopy.com 6, 25 -- From: Sergey {sryazant-at-ucla.edu} 6, 25 -- Subject: Re: [Microscopy] RE: TEM: Immunocytochemistry and Cu grids 6, 25 -- In-Reply-To: {200507131623.j6DGNPQ6024061-at-ns.microscopy.com} 6, 25 -- References: {200507131623.j6DGNPQ6024061-at-ns.microscopy.com} 6, 25 -- Mime-Version: 1.0 6, 25 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 6, 25 -- X-Probable-Spam: no 6, 25 -- X-Scanned-By: smtp.ucla.edu on 169.232.48.136 ==============================End of - Headers==============================
John Oren, Vermontoptech 802 425 2040 ----- Original Message ----- X-from: {andrew.mcnaughton-at-stonebow.otago.ac.nz} To: {micro-at-superlink.net} Sent: Wednesday, July 13, 2005 4:43 PM
Actually, TTU is only about 45 minutes from Plainview, and both the Dept. of Bio. Sci. and the TTU Medical School have electron microscopy labs.
-----Original Message----- X-from: gcc-at-couger.com [mailto:gcc-at-couger.com] Sent: Wednesday, July 13, 2005 1:05 PM To: jfb-at-uidaho.edu
Your best bet would be to call some one at Texas Tech in Lubbock http://www.macmed.ttuhsc.edu/ They may be more than an hour from Plainview but that close to the edge of the world. Your only other chance is Amarillo with the TAM Vet Med Diagnostic lab http://www.medcenter.org/facilities/am-vetlab.html.
Good luck Gordon
Gordon Couger I collect links on information related to light microscopes. www.couger.com/microscope/links/gclinks.html Please forward anything you think might be useful to others. Microscope Documentation is at www.science-info.org
tpepper-at-iastate.edu wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by } (tpepper-at-iastate.edu) from http://www.microscopy.org/MicroscopyListserver/MLFormMail.html } on Wednesday, July 13, 2005 at 09:21:59 } --------------------------------------------------------------------------- } } Email: tpepper-at-iastate.edu } Name: Tracey Pepper } } Organization: Iowa State University } } Title-Subject: [Filtered] MListserver: } } Question: HELP! We are in desparate need of some assisitance! We have a scientist in Texas who is in the field collecting crucial samples for TEM examination. They must be collected in the next couple of days. Our efforts to send (via DHL) good fixative (standard em fix for plants a double aldehyde in a 0.1 M cacodylate) have failed! The person is located in Plainview, Texas. Is there anyone on this listserver who could make us some fixtive (150 mls)for this person to come pick up? Preferably within an hour radius? If you can, Please call 515-294-3872 as soon as possible. We would be in your debt!! } With Great appreciation, } Tracey Pepper } Bessey Microscopy Facility } Iowa State University } Ames, IA 50011-1020 } } --------------------------------------------------------------------------- } } ==============================Original Headers============================== } 6, 12 -- From zaluzec-at-microscopy.com Wed Jul 13 13:54:01 2005 } 6, 12 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) } 6, 12 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6DIs1ei022223 } 6, 12 -- for {microscopy-at-microscopy.com} ; Wed, 13 Jul 2005 13:54:01 -0500 } 6, 12 -- Mime-Version: 1.0 } 6, 12 -- X-Sender: (Unverified) } 6, 12 -- Message-Id: {p06110400befb1371ac5b-at-[206.69.208.22]} } 6, 12 -- Date: Wed, 13 Jul 2005 13:54:00 -0500 } 6, 12 -- To: microscopy-at-microscopy.com } 6, 12 -- From: tpepper-at-iastate.edu (by way of MicroscopyListserver) } 6, 12 -- Subject: viaWWW: Help with Fixative near Plainview Texas } 6, 12 -- Content-Type: text/plain; charset="us-ascii" } ==============================End of - Headers============================== } }
==============================Original Headers============================== 5, 20 -- From gcc-at-couger.com Wed Jul 13 15:01:27 2005 5, 20 -- Received: from lakermmtao08.cox.net (lakermmtao08.cox.net [68.230.240.31]) 5, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6DK1RJr030657 5, 20 -- for {microscopy-at-microscopy.com} ; Wed, 13 Jul 2005 15:01:27 -0500 5, 20 -- Received: from [127.0.0.1] (really [68.12.43.168]) by lakermmtao08.cox.net 5, 20 -- (InterMail vM.6.01.04.00 201-2131-118-20041027) with ESMTP 5, 20 -- id {20050713200117.YBBU19415.lakermmtao08.cox.net-at-[127.0.0.1]} 5, 20 -- for {microscopy-at-microscopy.com} ; Wed, 13 Jul 2005 16:01:17 -0400 5, 20 -- Message-ID: {42D5730D.4090609-at-couger.com} 5, 20 -- Date: Wed, 13 Jul 2005 15:01:17 -0500 5, 20 -- From: Gordon Couger {gcc-at-couger.com} 5, 20 -- User-Agent: Mozilla Thunderbird 1.0.2 (Windows/20050317) 5, 20 -- X-Accept-Language: en-us, en 5, 20 -- MIME-Version: 1.0 5, 20 -- To: microscopy-at-microscopy.com 5, 20 -- Subject: Re: [Microscopy] viaWWW: Help with Fixative near Plainview Texas 5, 20 -- References: {200507131858.j6DIw5Mo027248-at-ns.microscopy.com} 5, 20 -- In-Reply-To: {200507131858.j6DIw5Mo027248-at-ns.microscopy.com} 5, 20 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 5, 20 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
==============================Original Headers============================== 14, 27 -- From jfb-at-uidaho.edu Wed Jul 13 16:27:33 2005 14, 27 -- Received: from proofagent.csrv.uidaho.edu (mx2.uidaho.edu [129.101.155.249]) 14, 27 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6DLRWRx029989 14, 27 -- for {microscopy-at-microscopy.com} ; Wed, 13 Jul 2005 16:27:32 -0500 14, 27 -- Received: from mailB.its.uidaho.edu (mailB.its.uidaho.edu [129.101.155.251]) 14, 27 -- by proofagent.csrv.uidaho.edu (8.13.3/8.13.3) with ESMTP id j6DLRVQW020697 14, 27 -- for {microscopy-at-microscopy.com} ; Wed, 13 Jul 2005 14:27:31 -0700 14, 27 -- Received: from jfb1 (PC006696.fs.uidaho.edu [129.101.141.111]) 14, 27 -- by mailB.its.uidaho.edu (Go Vandals!) 14, 27 -- with SMTP id {0IJL00CI44XVC6-at-mailB.its.uidaho.edu} for 14, 27 -- microscopy-at-microscopy.com; Wed, 13 Jul 2005 14:27:31 -0700 (PDT) 14, 27 -- Date: Wed, 13 Jul 2005 14:27:49 -0700 14, 27 -- From: Franklin Bailey {jfb-at-uidaho.edu} 14, 27 -- Subject: RE: [Microscopy] Re: viaWWW: Help with Fixative near Plainview Texas 14, 27 -- In-reply-to: {200507132004.j6DK4osn002913-at-ns.microscopy.com} 14, 27 -- To: microscopy-at-microscopy.com 14, 27 -- Message-id: {GPEIIPGKOJCCMLKMDOOFAEJDCLAA.jfb-at-uidaho.edu} 14, 27 -- MIME-version: 1.0 14, 27 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2800.1506 14, 27 -- X-Mailer: Microsoft Outlook IMO, Build 9.0.2416 (9.0.2911.0) 14, 27 -- Content-type: text/plain; charset=iso-8859-1 14, 27 -- Content-transfer-encoding: 7bit 14, 27 -- Importance: Normal 14, 27 -- X-Priority: 3 (Normal) 14, 27 -- X-MSMail-priority: Normal 14, 27 -- X-SpamDetails: rule=notspam policy=default score=0 mlx=0 adultscore=0 adjust=0 reason=mlx engine=3.0.0-05071100 definitions=3.0.0-05071302 14, 27 -- X-SpamScore: 0 ==============================End of - Headers==============================
Henk: I believe you are correct. The "hi-mag" mode does use an in-lens detector and you will get x-ray peaks you are not wanting due to the way the magnetic lens acts.
Larry: using "Analysis" mode will correct this problem. The WD for this scope with an INCA detector should be about 12mm +/- 1mm. Using the "hi-mag" mode for EDX work is not recommended; you won't get good resolution at this working distance anyway and the in-lens detector won't give a very good image at this distance, besides being very sensitive to charging.
colijn.1-at-osu.edu wrote:
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-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-648-8743 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
==============================Original Headers============================== 6, 23 -- From r-holdford-at-ti.com Wed Jul 13 17:39:30 2005 6, 23 -- Received: from go4.ext.ti.com (go4.ext.ti.com [192.91.75.132]) 6, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6DMdTS1005980 6, 23 -- for {microscopy-at-microscopy.com} ; Wed, 13 Jul 2005 17:39:30 -0500 6, 23 -- Received: from dlep30.itg.ti.com ([157.170.139.157]) 6, 23 -- by go4.ext.ti.com (8.13.1/8.13.1) with ESMTP id j6DMdN08019399 6, 23 -- for {microscopy-at-microscopy.com} ; Wed, 13 Jul 2005 17:39:28 -0500 (CDT) 6, 23 -- Received: from [156.117.194.45] (localhost [127.0.0.1]) 6, 23 -- by dlep30.itg.ti.com (8.12.11/8.12.11) with ESMTP id j6DMdN4b018550 6, 23 -- for {microscopy-at-microscopy.com} ; Wed, 13 Jul 2005 17:39:23 -0500 (CDT) 6, 23 -- Message-ID: {42D5981A.8050700-at-ti.com} 6, 23 -- Date: Wed, 13 Jul 2005 17:39:22 -0500 6, 23 -- From: Becky Holdford {r-holdford-at-ti.com} 6, 23 -- Organization: SC Packaging Development -- FA Development 6, 23 -- User-Agent: Mozilla Thunderbird 0.8 (Windows/20040913) 6, 23 -- X-Accept-Language: en-us, en 6, 23 -- MIME-Version: 1.0 6, 23 -- To: microscopy-at-microscopy.com 6, 23 -- Subject: Re: [Microscopy] Re: Stray X-rays in FE SEM? 6, 23 -- References: {200507130012.j6D0CZYa029297-at-ns.microscopy.com} 6, 23 -- In-Reply-To: {200507130012.j6D0CZYa029297-at-ns.microscopy.com} 6, 23 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 6, 23 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
The actual retail price of T221 in the summer of 2004 (when T221 was available from regular retail sources) was only $4K. Proper video adapters are $800 to $1,200, largely matter of taste. It is safe to say the whole deal was approximately. $5K. Totally worth it IMO. The figures around $8K to $10K discussed in this thread are MSRP, not retail prices.
Vitaly Feingold SIA 2773 Heath Lane Duluth, GA 30096 Ph. 770-232-7785 Fax 770-232-1791 www.sia-cam.com
----- Original Message ----- X-from: {Mike.Bode-at-soft-imaging.net} To: {vitalylazar-at-att.net} Sent: Friday, July 08, 2005 11:01 AM
Hi John,
The advantages besides the obvious superb display are:
1) the ability to display entire image at 100% zoom (pixel-to-pixel sensor-to-monitor) without zoom or pan;
2) have such image and multiple toolbars and control boxes displayed on the desktop without interfering with each other, plus an extra application or two open at the same time, again, not hiding behind other open windows- makes it much easier on the operator.
Works very well for 4 or 6 megapixels camera. Now, for larger sensor such as 11 or 16 megapixels, one would have to zoom out to 50% in order to see entire image at once. Still much better than zooming down to 25% on a regular monitor.
Vitaly Feingold SIA 2773 Heath Lane Duluth, GA 30096 Ph. 770-232-7785 Fax 770-232-1791 www.sia-cam.com
----- Original Message ----- X-from: {john.mardinly-at-intel.com} To: {vitalylazar-at-att.net} Sent: Friday, July 08, 2005 4:16 PM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (xuy-at-nih.gov) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Thursday, July 14, 2005 at 08:09:33 ---------------------------------------------------------------------------
Email: xuy-at-nih.gov Name: Yuhui Xu
Organization: Harvard Medical School
Title-Subject: [Filtered] MListserver:
Question: I am in the process of buying a digital camera for my light microscope ( Leica DM LB2), and would like to get your opinion as to which maker or model is a good choice in terms of resolution, stability, and ease of use, and of course the price.
Once again the MSA Education Committee has organized Exhibitor Demonstrations and Tutorials during the Microscopy and Microanalysis 2005 meeting. This will occur on Tuesday, August 2 starting at 5:00 pm in the Exhibit Hall. These mini-seminars and/or tutorial demonstrations are held in the booths of the participating companies after the Hall is closed to non-participants.
Reservations sheets with titles and descriptions will be at the MSA Education table in the MSA Mega Booth. When you sign up you will be issued a ticket, which you will need to renter the Hall after it is closed. You need to sign up no later than noon on Tuesday. The number of attendees is limited, so visit the MSA Education table soon, since the demonstrations get filled up quickly!
Here's a list of participating Exhibitors and titles:
Your problem is simply one of backscatter bouncing off the final lens irradiating areas off axis, generating the x-rays as appropriate. This is not just a problem with the SEM you mention, it is a general problem of which many people are unaware. The worst case I have ever seen was obtaining x-ray information from 0.75cm (~1/4inch) from the point of initial beam impact!
Modern detectors and collimators do help but scatter is always a problem. I believe that is why SEM manufacturer's main holder is often one which places the specimen surface at a higher level well away from the holder/stage interface. A specimen "in space" will always offer a cleaner x-ray signal.
It is a personal suggestion to clients that when trying hard with an analysis they DO NOT use a multi specimen holder!
Those who wish to know more should read texts relating to the production of SE in relation to those produced by "bounce off" backscatter. We have a short piece in the "Hints and Tips" section of our web site.
Steve Chapman Senior Consultant Protrain For electron microscopy consultancy and training world wide Tel +44 1280 816512 Fax +44 1280 814007 Mobile +44 7802 966067 Web www.emcourses.com
----- Original Message ----- X-from: {hanke-at-mee-inc.com} To: {protrain-at-emcourses.com} Sent: Wednesday, July 13, 2005 12:03 AM
We have an old Wild MTr19 illuminator for an equally old Wild model 20 compound light microscope. The bulb in the illuminator burnt out and from the sources I checked that type of bulb is supposedly no longer made. Also, there are no markings on the bulb to indicate what it is. Is anyone familiar with this old illuminator and either knows of bulb replacements or a compatible illuminator for which I can still get bulbs?
Thanks
Norm Olson ______________________________________________________________ Norm Olson Cryoelectron Microscopy Facilities Manager 4107 Natural Science Building Department of Chemistry & Biochemistry, MC-0378 University of California San Diego La Jolla, CA 92093-0378 nholson-at-ucsd.edu http://cryoem.ucsd.edu (858)534-5852 Office; (858)534-5846 - Fax ______________________________________________________________
==============================Original Headers============================== 9, 24 -- From nholson-at-ucsd.edu Thu Jul 14 13:52:19 2005 9, 24 -- Received: from mailbox4.ucsd.edu (mailbox4.ucsd.edu [132.239.1.56]) 9, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6EIqJbE032597 9, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 14 Jul 2005 13:52:19 -0500 9, 24 -- Received: from smtp.ucsd.edu (smtp-a.ucsd.edu [132.239.1.49]) 9, 24 -- by mailbox4.ucsd.edu (8.13.3/8.13.3) with ESMTP id j6EIOBhA036898 9, 24 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 9, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 14 Jul 2005 11:24:12 -0700 (PDT) 9, 24 -- Received: from [132.239.184.37] ([132.239.184.37]) 9, 24 -- by smtp.ucsd.edu (8.12.10/8.9.3) with ESMTP id j6EIO3Tg020814 9, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 14 Jul 2005 11:24:11 -0700 (PDT) 9, 24 -- User-Agent: Microsoft-Entourage/10.0.0.1309 9, 24 -- Date: Thu, 14 Jul 2005 11:24:03 -0700 9, 24 -- Subject: Illuminator for old Wild light microscope 9, 24 -- From: Norman Olson {nholson-at-ucsd.edu} 9, 24 -- To: {Microscopy-at-microscopy.com} 9, 24 -- Message-ID: {BEFBFBD3.28C6%nholson-at-ucsd.edu} 9, 24 -- Mime-version: 1.0 9, 24 -- Content-type: text/plain; charset="ISO-8859-1" 9, 24 -- X-Greylisting: NO DELAY (Trusted relay host); 9, 24 -- processed by UCSD_GL-v1.3 on mailbox4.ucsd.edu; 9, 24 -- Thu, 14 July 2005 11:24:12 -0700 (PDT) 9, 24 -- Content-Transfer-Encoding: 8bit 9, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id j6EIqJbE032597 ==============================End of - Headers==============================
Joe Mears knows everything you what to know about the Swiss made Wild. Microscope Services 1403 Bradley Avenue, Rockville, Maryland 20851 Office 301.294.7960 Fax 301.294.1934 Cell 240.994.7191
Joe also works on Leica, Zeiss, and Nikon light scopes. He is a very good repair person.
nholson-at-ucsd.edu wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } We have an old Wild MTr19 illuminator for an equally old Wild model 20 } compound light microscope. The bulb in the illuminator burnt out and from } the sources I checked that type of bulb is supposedly no longer made. Also, } there are no markings on the bulb to indicate what it is. Is anyone } familiar with this old illuminator and either knows of bulb replacements or } a compatible illuminator for which I can still get bulbs? } } Thanks } } Norm Olson } ______________________________________________________________ } Norm Olson } Cryoelectron Microscopy Facilities Manager } 4107 Natural Science Building } Department of Chemistry & Biochemistry, MC-0378 } University of California San Diego } La Jolla, CA 92093-0378 } nholson-at-ucsd.edu } http://cryoem.ucsd.edu } (858)534-5852 Office; (858)534-5846 - Fax } ______________________________________________________________ } } } } } } } } ==============================Original Headers============================== } 9, 24 -- From nholson-at-ucsd.edu Thu Jul 14 13:52:19 2005 } 9, 24 -- Received: from mailbox4.ucsd.edu (mailbox4.ucsd.edu [132.239.1.56]) } 9, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6EIqJbE032597 } 9, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 14 Jul 2005 13:52:19 -0500 } 9, 24 -- Received: from smtp.ucsd.edu (smtp-a.ucsd.edu [132.239.1.49]) } 9, 24 -- by mailbox4.ucsd.edu (8.13.3/8.13.3) with ESMTP id j6EIOBhA036898 } 9, 24 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) } 9, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 14 Jul 2005 11:24:12 -0700 (PDT) } 9, 24 -- Received: from [132.239.184.37] ([132.239.184.37]) } 9, 24 -- by smtp.ucsd.edu (8.12.10/8.9.3) with ESMTP id j6EIO3Tg020814 } 9, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 14 Jul 2005 11:24:11 -0700 (PDT) } 9, 24 -- User-Agent: Microsoft-Entourage/10.0.0.1309 } 9, 24 -- Date: Thu, 14 Jul 2005 11:24:03 -0700 } 9, 24 -- Subject: Illuminator for old Wild light microscope } 9, 24 -- From: Norman Olson {nholson-at-ucsd.edu} } 9, 24 -- To: {Microscopy-at-microscopy.com} } 9, 24 -- Message-ID: {BEFBFBD3.28C6%nholson-at-ucsd.edu} } 9, 24 -- Mime-version: 1.0 } 9, 24 -- Content-type: text/plain; charset="ISO-8859-1" } 9, 24 -- X-Greylisting: NO DELAY (Trusted relay host); } 9, 24 -- processed by UCSD_GL-v1.3 on mailbox4.ucsd.edu; } 9, 24 -- Thu, 14 July 2005 11:24:12 -0700 (PDT) } 9, 24 -- Content-Transfer-Encoding: 8bit } 9, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id j6EIqJbE032597 } ==============================End of - Headers============================== }
-- Chere Petty, M.S. Manager, Keith R. Porter Imaging Facility Department of Biological Sciences University of Maryland Baltimore County (UMBC) 1000 Hilltop Circle Baltimore, MD 21250 Phone: 410-455-2296 Fax: 410-455-3875
"The statements and opinions expressed here by Gary M. Brown represent neither those of ExxonMobil Corporation nor its affiliates."
Norm,
Leica should have a part number for the bulb and the vital stats needed to purchase it elsewhere.
Regards,
Gary M. Brown ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: 281 834 2387 fax: 281 834 2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
nholson-at-ucsd.e du To gary.m.brown-at-exxonmobil.com 07/14/05 01:54 cc PM Subject [Microscopy] Illuminator for old Please respond Wild light microscope to microscopy-at-mic roscopy.com
The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
We have an old Wild MTr19 illuminator for an equally old Wild model 20 compound light microscope. The bulb in the illuminator burnt out and from the sources I checked that type of bulb is supposedly no longer made. Also, there are no markings on the bulb to indicate what it is. Is anyone familiar with this old illuminator and either knows of bulb replacements or a compatible illuminator for which I can still get bulbs?
Thanks
Norm Olson ______________________________________________________________ Norm Olson Cryoelectron Microscopy Facilities Manager 4107 Natural Science Building Department of Chemistry & Biochemistry, MC-0378 University of California San Diego La Jolla, CA 92093-0378 nholson-at-ucsd.edu http://cryoem.ucsd.edu (858)534-5852 Office; (858)534-5846 - Fax ______________________________________________________________
==============================Original Headers============================== 9, 24 -- From nholson-at-ucsd.edu Thu Jul 14 13:52:19 2005 9, 24 -- Received: from mailbox4.ucsd.edu (mailbox4.ucsd.edu [132.239.1.56]) 9, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6EIqJbE032597 9, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 14 Jul 2005 13:52:19 -0500 9, 24 -- Received: from smtp.ucsd.edu (smtp-a.ucsd.edu [132.239.1.49]) 9, 24 -- by mailbox4.ucsd.edu (8.13.3/8.13.3) with ESMTP id j6EIOBhA036898 9, 24 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 9, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 14 Jul 2005 11:24:12 -0700 (PDT) 9, 24 -- Received: from [132.239.184.37] ([132.239.184.37]) 9, 24 -- by smtp.ucsd.edu (8.12.10/8.9.3) with ESMTP id j6EIO3Tg020814 9, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 14 Jul 2005 11:24:11 -0700 (PDT) 9, 24 -- User-Agent: Microsoft-Entourage/10.0.0.1309 9, 24 -- Date: Thu, 14 Jul 2005 11:24:03 -0700 9, 24 -- Subject: Illuminator for old Wild light microscope 9, 24 -- From: Norman Olson {nholson-at-ucsd.edu} 9, 24 -- To: {Microscopy-at-microscopy.com} 9, 24 -- Message-ID: {BEFBFBD3.28C6%nholson-at-ucsd.edu} 9, 24 -- Mime-version: 1.0 9, 24 -- Content-type: text/plain; charset="ISO-8859-1" 9, 24 -- X-Greylisting: NO DELAY (Trusted relay host); 9, 24 -- processed by UCSD_GL-v1.3 on mailbox4.ucsd.edu; 9, 24 -- Thu, 14 July 2005 11:24:12 -0700 (PDT) 9, 24 -- Content-Transfer-Encoding: 8bit 9, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id j6EIqJbE032597 ==============================End of - Headers==============================
==============================Original Headers============================== 30, 21 -- From gary.m.brown-at-exxonmobil.com Thu Jul 14 16:37:49 2005 30, 21 -- Received: from hoespc02.exxonmobil.com (hoespc02.exxonmobil.com [192.67.48.39]) 30, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6ELbnYQ017505 30, 21 -- for {microscopy-at-microscopy.com} ; Thu, 14 Jul 2005 16:37:49 -0500 30, 21 -- Received: from hounmg03.NA.XOM.COM (hounmg03.na.xom.com [158.35.101.41]) 30, 21 -- by hoespc02.exxonmobil.com (Switch-3.1.7/Switch-3.1.7) with ESMTP id j6ELbkCx010176 30, 21 -- for {microscopy-at-microscopy.com} ; Thu, 14 Jul 2005 16:37:49 -0500 (CDT) 30, 21 -- In-Reply-To: {200507141854.j6EIsqtp004351-at-ns.microscopy.com} 30, 21 -- Subject: Re: [Microscopy] Illuminator for old Wild light microscope 30, 21 -- Importance: 30, 21 -- To: microscopy-at-microscopy.com 30, 21 -- X-Mailer: Lotus Notes Release 6.5.2 June 01, 2004 30, 21 -- Message-ID: {OF0700E947.8B6F5623-ON8625703E.007675EB-8625703E.0076D0A3-at-exxonmobil.com} 30, 21 -- From: gary.m.brown-at-exxonmobil.com 30, 21 -- Date: Thu, 14 Jul 2005 16:37:46 -0500 30, 21 -- X-MIMETrack: Serialize by Router on Hounmg03.na.xom.com/S/ExxonMobil(652HF702|December 30, 21 -- 14, 2004) at 07/14/2005 04:37:49 PM 30, 21 -- MIME-Version: 1.0 30, 21 -- Content-type: text/plain; charset=ISO-8859-1 30, 21 -- Content-Transfer-Encoding: 8bit 30, 21 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id j6ELbnYQ017505 ==============================End of - Headers==============================
Does anyone know of a company that can service an old Gatan Duomill? Ours has developed an electrical problem that keeps the guns from energizing.
Thanks in advance.
----------------------------------------- John Bonevich, Ph.D. National Institute of Standards and Technology Metallurgy Division, Mail-Stop 8555 100 Bureau Drive Gaithersburg, MD 20899 USA
==============================Original Headers============================== 5, 20 -- From john.bonevich-at-nist.gov Fri Jul 15 12:11:10 2005 5, 20 -- Received: from smtp.nist.gov (rimp2.nist.gov [129.6.16.227]) 5, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6FHBADR005599 5, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 15 Jul 2005 12:11:10 -0500 5, 20 -- Received: from postmark.nist.gov (pullyou.nist.gov [129.6.16.93]) 5, 20 -- by smtp.nist.gov (8.13.1/8.13.1) with ESMTP id j6FHB7S3009790 5, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 15 Jul 2005 13:11:07 -0400 5, 20 -- Received: from h180168.nist.gov (h180168.nist.gov [129.6.180.168]) 5, 20 -- by postmark.nist.gov (8.12.5/8.12.5) with ESMTP id j6FHAO6v002771 5, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 15 Jul 2005 13:10:24 -0400 (EDT) 5, 20 -- Message-Id: {6.2.1.2.2.20050715130621.0389b1e0-at-email.nist.gov} 5, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.1.2 5, 20 -- Date: Fri, 15 Jul 2005 13:10:21 -0400 5, 20 -- To: Microscopy-at-MSA.Microscopy.Com 5, 20 -- From: John Bonevich {john.bonevich-at-nist.gov} 5, 20 -- Subject: Gatan Duomill service 5, 20 -- Mime-Version: 1.0 5, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 5, 20 -- X-NIST-MailScanner: Found to be clean 5, 20 -- X-NIST-MailScanner-From: john.bonevich-at-nist.gov ==============================End of - Headers==============================
-----Original Message----- X-from: john.bonevich-at-nist.gov [mailto:john.bonevich-at-nist.gov] Sent: Friday, July 15, 2005 10:15 AM To: curulli-at-usc.edu
Hello,
Does anyone know of a company that can service an old Gatan Duomill? Ours has developed an electrical problem that keeps the guns from energizing.
Thanks in advance.
----------------------------------------- John Bonevich, Ph.D. National Institute of Standards and Technology Metallurgy Division, Mail-Stop 8555 100 Bureau Drive Gaithersburg, MD 20899 USA
==============================Original Headers============================== 5, 20 -- From john.bonevich-at-nist.gov Fri Jul 15 12:11:10 2005 5, 20 -- Received: from smtp.nist.gov (rimp2.nist.gov [129.6.16.227]) 5, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6FHBADR005599 5, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 15 Jul 2005 12:11:10 -0500 5, 20 -- Received: from postmark.nist.gov (pullyou.nist.gov [129.6.16.93]) 5, 20 -- by smtp.nist.gov (8.13.1/8.13.1) with ESMTP id j6FHB7S3009790 5, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 15 Jul 2005 13:11:07 -0400 5, 20 -- Received: from h180168.nist.gov (h180168.nist.gov [129.6.180.168]) 5, 20 -- by postmark.nist.gov (8.12.5/8.12.5) with ESMTP id j6FHAO6v002771 5, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 15 Jul 2005 13:10:24 -0400 (EDT) 5, 20 -- Message-Id: {6.2.1.2.2.20050715130621.0389b1e0-at-email.nist.gov} 5, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.1.2 5, 20 -- Date: Fri, 15 Jul 2005 13:10:21 -0400 5, 20 -- To: Microscopy-at-MSA.Microscopy.Com 5, 20 -- From: John Bonevich {john.bonevich-at-nist.gov} 5, 20 -- Subject: Gatan Duomill service 5, 20 -- Mime-Version: 1.0 5, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 5, 20 -- X-NIST-MailScanner: Found to be clean 5, 20 -- X-NIST-MailScanner-From: john.bonevich-at-nist.gov ==============================End of - Headers==============================
==============================Original Headers============================== 14, 20 -- From curulli-at-usc.edu Fri Jul 15 12:43:29 2005 14, 20 -- Received: from msg-mx5.usc.edu (msg-mx5.usc.edu [128.125.137.10]) 14, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6FHhSes013552 14, 20 -- for {microscopy-at-microscopy.com} ; Fri, 15 Jul 2005 12:43:28 -0500 14, 20 -- Received: from CEMMA2 ([128.125.153.147]) 14, 20 -- by msg-mx5.usc.edu (Sun Java System Messaging Server 6.2-2.07 (built May 25 14, 20 -- 2005)) with ESMTP id {0IJO0055WJWDOOA0-at-msg-mx5.usc.edu} for 14, 20 -- microscopy-at-microscopy.com; Fri, 15 Jul 2005 10:43:28 -0700 (PDT) 14, 20 -- Date: Fri, 15 Jul 2005 10:43:24 -0700 14, 20 -- From: John Curulli {curulli-at-usc.edu} 14, 20 -- Subject: RE: [Microscopy] Gatan Duomill service 14, 20 -- In-reply-to: {200507151714.j6FHEWHj009497-at-ns.microscopy.com} 14, 20 -- To: microscopy-at-microscopy.com 14, 20 -- Message-id: {0IJO0055ZJWGOOA0-at-msg-mx5.usc.edu} 14, 20 -- MIME-version: 1.0 14, 20 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 14, 20 -- X-Mailer: Microsoft Office Outlook, Build 11.0.6353 14, 20 -- Content-type: text/plain; charset=us-ascii 14, 20 -- Content-transfer-encoding: 7BIT 14, 20 -- Thread-index: AcWJYKzdElCIuMriSBCO+fa/tcHJtQAA9csA ==============================End of - Headers==============================
I am an amateur and have been experimenting for quite some time with a technique for sharpening silicon carbide (SiC) crystals as an alternative to the very expensive diamond knives commonly used with ultramicrotomes.
The major difficulty is that SiC crystals are very brittle; thus SiC crystals need to be lapped and polished by unconventional methods to prevent micro-mechanical stresses from breaking the edge. Conventional gem faceting technology does not work.
Vitreous carbon knives have also been reported in the literature but did not live up to expectations apparently.
As many on this list will know, glass knives are preferred for their initial sharpness. Meanwhile diamond knives are preferred for their unexcelled durability, flat faces and straight edges, which makes it possible to make almost unlimited numbers of serial sections up to a few millimeters wide.
My efforts indicate that it is possible to make SiC knives and that they can generate silvery sections down to 100 nanometers or so, which puts them in the ball park for practical EM work. My sections do transmit electrons, and at best do not appear to be much worse than diamond knives in regards to their initial sharpness.
Initially I had imagined that SiC knives might potentially replace diamond knives, since the hardness of SiC crystals is 9+ on the Mho's scale, while diamond is 10.
As it turns out, my SiC knives gradually become dull over the course of making hundreds of sections. However they are distinctly more durable than glass knives, and the lapped faces are much flatter and the edges wider compared to glass.
The SiC knife durability seems to be a function of the hardness of the embedding plastic being sectioned, as one may imagine. (They section glycol methacrylate nicely, for making floating ribbons, which is helpful since I intend to use the SiC knives myself for LM work).
My question is how useful such knives are likely to be in practice? I imagine SiC knives might fill a niche market for disposable substitutes for diamond knives, when an inexpensive alternative to glass knives is needed only occasionally and the purchase of a diamond knife doesn't make sense, or maybe for student teaching.
It seems likely that the same technology can probably be used to make sapphire knives of equivalent initial sharpness (sapphire is much tougher and more fracture resistant than SiC but not so hard as SiC). But I don't know, my efforts so far have only concerned SiC knives.
I expect to publish my SiC knife making technology and put it in the public domain, but for now I am soliciting comments on their potential usefulness. -- Roger, Austin, Tx
==============================Original Headers============================== 17, 19 -- From rcbaker-at-eden.infohwy.com Sat Jul 16 10:57:37 2005 17, 19 -- Received: from mx2.lsn.net (mx2.lsn.net [66.90.130.74]) 17, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6GFvbcC004113 17, 19 -- for {microscopy-at-microscopy.com} ; Sat, 16 Jul 2005 10:57:37 -0500 17, 19 -- Received: from [192.168.1.100] (66-90-146-134.dyn.grandenetworks.net [66.90.146.134]) 17, 19 -- by mx2.lsn.net (8.13.0.Beta3/8.12.8) with ESMTP id j6GFvZef022611 17, 19 -- for {microscopy-at-microscopy.com} ; Sat, 16 Jul 2005 10:57:40 -0500 17, 19 -- Mime-Version: 1.0 (Apple Message framework v730) 17, 19 -- In-Reply-To: {200507151716.j6FHGuR9011532-at-ns.microscopy.com} 17, 19 -- References: {200507151716.j6FHGuR9011532-at-ns.microscopy.com} 17, 19 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 17, 19 -- Message-Id: {472D4ED4-B5BB-474E-BDF9-BEE268C8A16A-at-eden.infohwy.com} 17, 19 -- Content-Transfer-Encoding: 7bit 17, 19 -- From: Roger Baker {rcbaker-at-eden.infohwy.com} 17, 19 -- Subject: Alternatives to diamond knives? 17, 19 -- Date: Sat, 16 Jul 2005 10:57:25 -0500 17, 19 -- To: microscopy-at-microscopy.com 17, 19 -- X-Mailer: Apple Mail (2.730) 17, 19 -- X-Antivirus: Scanned by Vexira Antivirus 1.0.6 ==============================End of - Headers==============================
Contact Art McCanna at MAS amccanna-at-mastest.com ; (770)866-3200; direct-(770)866-3212. MAS is exclusive factory representative for GATAN ion mills.
Vitaly Feingold SIA 2773 Heath Lane Duluth, GA 30096 Ph. 770-232-7785 Fax 770-232-1791 www.sia-cam.com
----- Original Message ----- X-from: {john.bonevich-at-nist.gov} To: {vitalylazar-at-att.net} Sent: Friday, July 15, 2005 1:13 PM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (luce-at-earthtech.org) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Sunday, July 17, 2005 at 16:06:51 ---------------------------------------------------------------------------
Email: luce-at-earthtech.org Name: George Luce
Organization: Earthtech Internaional, Inc.
Education: Graduate College
Location: Austin, Texas, USA
Question: We are looking for the electrical schematics for an International Scientific Instruments (ISI) Model 100B Scanning Electron Microscope. (~1979 vintage, probably made by Akashi)
Is this company still in business, or is there another resource for information about this SEM?
Thanks,
George Luce luce-at-earthtech.org
Earthtech International Inc. Austin, TX www.earthtech.org
the only comment that I can make is that sapphire knives were marketed a few (maybe 20) years ago but they were much more expensive than glass and not as hard as diamond so were never really that popular. I suppose in the real world a lot will depend on price and quality and whether there is a sufficiently large market, now.
I wish you luck.
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 ----- X-from: rcbaker-at-eden.infohwy.com
Hi Roger, It sounds interesting, keep us informed. Years ago, Diatome did market a Sapphire knife as a less-expensive alternative to diamonds. In my lab, they proved difficult to clean, and did not wear well enough to continue using them. They seem to have disappeared from the market, so my guess is that others had the same observations. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
==============================Original Headers============================== 1, 23 -- From lcgould-at-med.cornell.edu Mon Jul 18 09:11:25 2005 1, 23 -- Received: from smtp-in1.med.cornell.edu (smtp-in1.med.cornell.edu [140.251.1.25]) 1, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6IEBPjw018439 1, 23 -- for {microscopy-at-microscopy.com} ; Mon, 18 Jul 2005 09:11:25 -0500 1, 23 -- Received: from mpx2.med.cornell.edu (pc113142-10.med.cornell.edu [140.251.11.119]) 1, 23 -- by smtp-in1.med.cornell.edu (Switch-3.1.6/Switch-3.1.6) with ESMTP id j6IEBNmQ207152 1, 23 -- for {microscopy-at-microscopy.com} ; Mon, 18 Jul 2005 10:11:23 -0400 1, 23 -- Received: from [140.251.145.131] by mpx2.med.cornell.edu 1, 23 -- (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 28 2005)) 1, 23 -- with ESMTP id {0IJT00CEIU2Y6S90-at-mpx2.med.cornell.edu} for 1, 23 -- microscopy-at-microscopy.com; Mon, 18 Jul 2005 10:11:23 -0400 (EDT) 1, 23 -- Date: Mon, 18 Jul 2005 10:11:20 -0400 1, 23 -- From: Leona Cohen-Gould {lcgould-at-med.cornell.edu} 1, 23 -- Subject: Re: [Microscopy] Alternatives to diamond knives? 1, 23 -- In-reply-to: {200507161559.j6GFxljV005537-at-ns.microscopy.com} 1, 23 -- X-Sender: lcgould-at-pop.med.cornell.edu 1, 23 -- To: microscopy-at-microscopy.com 1, 23 -- Cc: rcbaker-at-eden.infohwy.com 1, 23 -- Message-id: {p0602040abf01684f55e8-at-[140.251.145.131]} 1, 23 -- MIME-version: 1.0 1, 23 -- Content-type: text/plain; charset=us-ascii; format=flowed 1, 23 -- References: {200507161559.j6GFxljV005537-at-ns.microscopy.com} 1, 23 -- X-PMX-Version: 4.7.1.128075, Antispam-Engine: 2.0.3.2, Antispam-Data: 2005.7.18.11 ==============================End of - Headers==============================
The problem has been solved by using external fans running through tubes to the camera.
Roper and Cooke each have their own different designs for the fan. Each were extremely helpful with the retrofits.
Cooke runs the camera power through the fan; this assures that the fan must be on for the camera to run. Roper doesn't have this safety feature.
I would recommend not purchasing a camera with an on board fan. Always go for external cooling.
Thanks for the help from everybody who responded to my query ____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/ **This electronic transmission contains information that is privileged.**
==============================Original Headers============================== 6, 24 -- From cammer-at-aecom.yu.edu Mon Jul 18 09:14:33 2005 6, 24 -- Received: from mailgw.aecom.yu.edu (mailgw.aecom.yu.edu [129.98.1.16]) 6, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6IEEXZ0022691 6, 24 -- for {microscopy-at-msa.microscopy.com} ; Mon, 18 Jul 2005 09:14:33 -0500 6, 24 -- Received: from mailvx.aecom.yu.edu (mailvx.aecom.yu.edu [129.98.1.17]) 6, 24 -- by mailgw.aecom.yu.edu (8.12.11/8.12.11) with SMTP id j6IEE9B9030533 6, 24 -- for {microscopy-at-msa.microscopy.com} ; Mon, 18 Jul 2005 10:14:33 -0400 6, 24 -- Received: from post.aecom.yu.edu ([129.98.1.100]) 6, 24 -- by mailvx.aecom.yu.edu (SAVSMTP 3.1.1.32) with SMTP id M2005071810143320737 6, 24 -- for {microscopy-at-msa.microscopy.com} ; Mon, 18 Jul 2005 10:14:33 -0400 6, 24 -- Received: from AIF3.aecom.yu.edu (aif3.aif.aecom.yu.edu [129.98.30.137]) 6, 24 -- by post.aecom.yu.edu (Postfix) with ESMTP id E869A2FC0 6, 24 -- for {microscopy-at-msa.microscopy.com} ; Mon, 18 Jul 2005 10:14:32 -0400 (EDT) 6, 24 -- Message-Id: {5.2.1.1.2.20050718100956.033d4060-at-mailserver.aecom.yu.edu} 6, 24 -- X-Sender: cammer-at-mailserver.aecom.yu.edu 6, 24 -- X-Mailer: QUALCOMM Windows Eudora Version 5.2.1 6, 24 -- Date: Mon, 18 Jul 2005 10:14:37 -0400 6, 24 -- To: microscopy-at-msa.microscopy.com 6, 24 -- From: Michael Cammer {cammer-at-aecom.yu.edu} 6, 24 -- Subject: Re: vibrations in air cooled CCD cameras; follow-up 6, 24 -- In-Reply-To: {20050525210830.20166.qmail-at-web81301.mail.yahoo.com} 6, 24 -- References: {6667} 6, 24 -- Mime-Version: 1.0 6, 24 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
I have a student who wants to look at clusters of methanosarcina and methanosaeta in an aqueous solution. I have a materials background and have never worked with this type of sample. The student found a reference for preparing samples for electron microscopy which looks to me like it is for TEM. She is interested in what the clusters look like so is interested in retaining their relative position. I think SEM would be a good technique to see the 3D cluster if there is a method to prepare samples.
I have a FESEM which is high vacuum. I don't have easy access to an environmental SEM, so I'm looking for a technique to image these clusters in a high vacuum SEM.
Any suggestions for prep techniques and/or references?
Thanks,
Tom
------------------------------------------------------------------------ --------------------------------------------- Thomas M Murray email: murraytm-at-u.washington.edu Electron Microscopy Center Manager Phone: (206)543-2836 Materials Science & Engineering Fax: (206)543-3100 Box 352120 302 Roberts Hall Cell: (425)345-0083 University of Washington Seattle, WA 98195
==============================Original Headers============================== 7, 24 -- From murraytm-at-u.washington.edu Mon Jul 18 10:56:23 2005 7, 24 -- Received: from mxout1.cac.washington.edu (mxout1.cac.washington.edu [140.142.32.134]) 7, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6IFuMul003018 7, 24 -- for {microscopy-at-microscopy.com} ; Mon, 18 Jul 2005 10:56:22 -0500 7, 24 -- Received: from smtp.washington.edu (smtp.washington.edu [140.142.32.139]) 7, 24 -- by mxout1.cac.washington.edu (8.13.4+UW05.04/8.13.4+UW05.05) with ESMTP id j6IFuLsd018291 7, 24 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=OK) 7, 24 -- for {microscopy-at-microscopy.com} ; Mon, 18 Jul 2005 08:56:21 -0700 7, 24 -- Received: from [128.95.118.89] (tstoebe-mse.ad.engr.washington.edu [128.95.118.89]) 7, 24 -- (authenticated authid=murraytm) 7, 24 -- by smtp.washington.edu (8.13.4+UW05.04/8.13.4+UW05.05) with ESMTP id j6IFuLaF015330 7, 24 -- (version=TLSv1/SSLv3 cipher=RC4-SHA bits=128 verify=NOT) 7, 24 -- for {microscopy-at-microscopy.com} ; Mon, 18 Jul 2005 08:56:21 -0700 7, 24 -- Mime-Version: 1.0 (Apple Message framework v733) 7, 24 -- In-Reply-To: {200507141333.j6EDXIb0014709-at-ns.microscopy.com} 7, 24 -- References: {200507141333.j6EDXIb0014709-at-ns.microscopy.com} 7, 24 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 7, 24 -- Message-Id: {9EC9D803-ACF3-4069-AA98-E78D3ABBACB2-at-u.washington.edu} 7, 24 -- Content-Transfer-Encoding: 7bit 7, 24 -- From: Tom Murray {murraytm-at-u.washington.edu} 7, 24 -- Subject: SEM Sample Prep Question 7, 24 -- Date: Mon, 18 Jul 2005 08:56:19 -0700 7, 24 -- To: microscopy-at-microscopy.com 7, 24 -- X-Mailer: Apple Mail (2.733) ==============================End of - Headers==============================
Al Coritz, Technical Engineer Electron Microscopy Sciences www.emsdiasum.com
----- Original Message ----- X-from: {murraytm-at-u.washington.edu} To: {Sampleprep-at-earthlink.net} Sent: Monday, July 18, 2005 11:56 AM
A colleague of mine is looking for an intensive TEM short course, 3-4 days, that covers theory and operation of TEM's, as well as interpretation of images, particularly as they are used in materials science. The course at Lehigh University for this year was just held, and waiting until next year is not a good option.
If you teach or know about such a course, please contact me offline and I will it forward the information.
Thanks very much,
--John Chandler Fort Collins, CO jchandler-at-ial-fa.com 970.217.1321
==============================Original Headers============================== 5, 22 -- From jchandler-at-ial-fa.com Mon Jul 18 11:51:41 2005 5, 22 -- Received: from sole.serverhost.net (sole.serverhost.net [216.71.84.188]) 5, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6IGpfq9019267 5, 22 -- for {Microscopy-at-microscopy.com} ; Mon, 18 Jul 2005 11:51:41 -0500 5, 22 -- Received: from JohnChandler (proxy1.lsil.com [147.145.40.41]) 5, 22 -- by sole.serverhost.net (8.11.6/8.11.6) with SMTP id j6IGpeg09884 5, 22 -- for {Microscopy-at-microscopy.com} ; Mon, 18 Jul 2005 12:51:40 -0400 5, 22 -- Message-ID: {004c01c58bb8$d6f9ddf0$02000000-at-JohnChandler} 5, 22 -- Reply-To: "John Chandler" {jchandler-at-ial-fa.com} 5, 22 -- From: "John Chandler" {jchandler-at-ial-fa.com} 5, 22 -- To: {Microscopy-at-microscopy.com} 5, 22 -- Subject: Training: looking for TEM short course 5, 22 -- Date: Mon, 18 Jul 2005 10:50:43 -0600 5, 22 -- Organization: Insight Analytical Labs 5, 22 -- MIME-Version: 1.0 5, 22 -- Content-Type: text/plain; 5, 22 -- charset="iso-8859-1" 5, 22 -- Content-Transfer-Encoding: 7bit 5, 22 -- X-Priority: 3 5, 22 -- X-MSMail-Priority: Normal 5, 22 -- X-Mailer: Microsoft Outlook Express 6.00.2800.1506 5, 22 -- x-mimeole: Produced By Microsoft MimeOLE V6.00.2800.1506 ==============================End of - Headers==============================
What's the procedure? Very likely, she can use it up through the 100% ethanol steps, whether or not it is for TEM. The critters can be filtered onto 0.22 micron membrane filters (the kind with nice, round pores, not a Millipore), and the filter critical point dried. An alternative to CPD is to air dry from HMDS (hexamethyldisilizane). Process the samples in 1.5 mL minifuge tubes, not on filters. After the last EtOH, go through a 2:1 1:1 1:2 EtOH:HMDS series, 3 changes in 100% HMDS, put sputter coated filters on the SEM stubs, drop the bugs in HMDS on the filters, and allow to air dry at room temp **do all this in a fume hood!**. See the U. Florida "tips and tricks" web site: http://www.biotech.ufl.edu/EM/tips/index.html
Or have a chat with the U. Washington biological EM people.
Phil
} I have a student who wants to look at clusters of methanosarcina and } methanosaeta in an aqueous solution. I have a materials background } and have never worked with this type of sample. The student found a } reference for preparing samples for electron microscopy which looks } to me like it is for TEM. She is interested in what the clusters } look like so is interested in retaining their relative position. I } think SEM would be a good technique to see the 3D cluster if there is } a method to prepare samples. } } I have a FESEM which is high vacuum. I don't have easy access to an } environmental SEM, so I'm looking for a technique to image these } clusters in a high vacuum SEM. } } Any suggestions for prep techniques and/or references? } } Thanks, } } Tom } } ------------------------------------------------------------------------ } --------------------------------------------- } Thomas M Murray email: } murraytm-at-u.washington.edu } Electron Microscopy Center Manager Phone: (206)543-2836 } Materials Science & Engineering Fax: (206)543-3100 } Box 352120 302 Roberts Hall Cell: (425)345-0083 } University of Washington } Seattle, WA 98195 } } } ==============================Original Headers============================== } 7, 24 -- From murraytm-at-u.washington.edu Mon Jul 18 10:56:23 2005 } 7, 24 -- Received: from mxout1.cac.washington.edu } (mxout1.cac.washington.edu [140.142.32.134]) } 7, 24 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id } j6IFuMul003018 } 7, 24 -- for {microscopy-at-microscopy.com} ; Mon, 18 Jul 2005 } 10:56:22 -0500 } 7, 24 -- Received: from smtp.washington.edu (smtp.washington.edu } [140.142.32.139]) } 7, 24 -- by mxout1.cac.washington.edu } (8.13.4+UW05.04/8.13.4+UW05.05) with ESMTP id j6IFuLsd018291 } 7, 24 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA } bits=256 verify=OK) } 7, 24 -- for {microscopy-at-microscopy.com} ; Mon, 18 Jul 2005 } 08:56:21 -0700 } 7, 24 -- Received: from [128.95.118.89] } (tstoebe-mse.ad.engr.washington.edu [128.95.118.89]) } 7, 24 -- (authenticated authid=murraytm) } 7, 24 -- by smtp.washington.edu } (8.13.4+UW05.04/8.13.4+UW05.05) with ESMTP id j6IFuLaF015330 } 7, 24 -- (version=TLSv1/SSLv3 cipher=RC4-SHA bits=128 verify=NOT) } 7, 24 -- for {microscopy-at-microscopy.com} ; Mon, 18 Jul 2005 } 08:56:21 -0700 } 7, 24 -- Mime-Version: 1.0 (Apple Message framework v733) } 7, 24 -- In-Reply-To: {200507141333.j6EDXIb0014709-at-ns.microscopy.com} } 7, 24 -- References: {200507141333.j6EDXIb0014709-at-ns.microscopy.com} } 7, 24 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed } 7, 24 -- Message-Id: {9EC9D803-ACF3-4069-AA98-E78D3ABBACB2-at-u.washington.edu} } 7, 24 -- Content-Transfer-Encoding: 7bit } 7, 24 -- From: Tom Murray {murraytm-at-u.washington.edu} } 7, 24 -- Subject: SEM Sample Prep Question } 7, 24 -- Date: Mon, 18 Jul 2005 08:56:19 -0700 } 7, 24 -- To: microscopy-at-microscopy.com } 7, 24 -- X-Mailer: Apple Mail (2.733) } ==============================End of - Headers==============================
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) http://www.ansci.wisc.edu/microscopy.htm
==============================Original Headers============================== 6, 27 -- From peoshel-at-wisc.edu Mon Jul 18 12:50:56 2005 6, 27 -- Received: from smtp7.wiscmail.wisc.edu (hagen.doit.wisc.edu [144.92.197.163]) 6, 27 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6IHounE027818 6, 27 -- for {microscopy-at-microscopy.com} ; Mon, 18 Jul 2005 12:50:56 -0500 6, 27 -- Received: from avs-daemon.smtp7.wiscmail.wisc.edu by smtp7.wiscmail.wisc.edu 6, 27 -- (iPlanet Messaging Server 5.2 HotFix 2.04 (built Feb 8 2005)) 6, 27 -- id {0IJU00F5348SA4-at-smtp7.wiscmail.wisc.edu} for microscopy-at-microscopy.com; 6, 27 -- Mon, 18 Jul 2005 12:50:52 -0500 (CDT) 6, 27 -- Received: from [10.25.102.29] (ansci.wisc.edu [144.92.132.175]) 6, 27 -- by smtp7.wiscmail.wisc.edu 6, 27 -- (iPlanet Messaging Server 5.2 HotFix 2.04 (built Feb 8 2005)) 6, 27 -- with ESMTPSA id {0IJU0085948QH8-at-smtp7.wiscmail.wisc.edu} for 6, 27 -- microscopy-at-microscopy.com; Mon, 18 Jul 2005 12:50:51 -0500 (CDT) 6, 27 -- Date: Mon, 18 Jul 2005 12:50:50 -0500 6, 27 -- From: Philip Oshel {peoshel-at-wisc.edu} 6, 27 -- Subject: Re: [Microscopy] SEM Sample Prep Question 6, 27 -- In-reply-to: {200507181557.j6IFvBgC004474-at-ns.microscopy.com} 6, 27 -- X-Sender: peoshel-at-wiscmail.wisc.edu 6, 27 -- To: microscopy-at-microscopy.com 6, 27 -- Message-id: {p05210601bf019a009786-at-[10.25.102.29]} 6, 27 -- MIME-version: 1.0 6, 27 -- Content-type: text/plain; format=flowed; charset=us-ascii 6, 27 -- Content-transfer-encoding: 7BIT 6, 27 -- X-Spam-Report: AuthenticatedSender=yes, SenderIP=144.92.132.175 6, 27 -- X-Spam-PmxInfo: Server=avs-2, Version=4.7.1.128075, Antispam-Engine: 2.0.3.1, 6, 27 -- Antispam-Data: 2005.7.18.20, SenderIP=144.92.132.175 6, 27 -- References: {200507181557.j6IFvBgC004474-at-ns.microscopy.com} ==============================End of - Headers==============================
One potential use would be to section materials that might damage a diamond knife, but that could not be cut using a glass knife. For example, specimens containing potentially damaging inclusions as sand, metals, etc. or to cut hard botanical specimens. -- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Email: bozzola-at-siu.edu Web: http://www.siu.edu/~image/ ##############################################################
==============================Original Headers============================== 2, 18 -- From bozzola-at-siu.edu Mon Jul 18 14:49:53 2005 2, 18 -- Received: from abbmta2.siu.edu (abbmta2.siu.edu [131.230.254.206]) 2, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6IJnrxe004719 2, 18 -- for {microscopy-at-microscopy.com} ; Mon, 18 Jul 2005 14:49:53 -0500 2, 18 -- Received: from [131.230.177.142] (ws177142.microscope.siu.edu [131.230.177.142]) 2, 18 -- by abbmta2.siu.edu (Switch-3.1.6/Switch-3.1.6) with ESMTP id j6IJnpvo026923 2, 18 -- for {microscopy-at-microscopy.com} ; Mon, 18 Jul 2005 14:49:52 -0500 (CDT) 2, 18 -- Mime-Version: 1.0 2, 18 -- X-Sender: bozzola-at-saluki-mail.siu.edu 2, 18 -- Message-Id: {p06110400bf01b80ddb8f-at-[131.230.177.142]} 2, 18 -- In-Reply-To: {200507161600.j6GG0kUa006196-at-ns.microscopy.com} 2, 18 -- References: {200507161600.j6GG0kUa006196-at-ns.microscopy.com} 2, 18 -- Date: Mon, 18 Jul 2005 14:49:50 -0500 2, 18 -- To: microscopy-at-microscopy.com 2, 18 -- From: "John J. Bozzola" {bozzola-at-siu.edu} 2, 18 -- Subject: Re: [Microscopy] Alternatives to diamond knives? 2, 18 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 2, 18 -- X-MASF: 0.00% ==============================End of - Headers==============================
We have a Winter School from January 9 through 12. One half day on the 13th. Please go to our web site where you will find a description of last year's school. We will have the info for 2006 up in a few days. Let me know if you need more info. John Wheatley
} ---------- } From: jchandler-at-ial-fa.com } Reply To: microscopy-at-microscopy.com } Sent: Monday, July 18, 2005 9:53 AM } To: John Wheatley } Subject: [Microscopy] Training: looking for TEM short course } } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } A colleague of mine is looking for an intensive TEM short course, 3-4 days, } that covers theory and operation of TEM's, as well as interpretation of } images, particularly as they are used in materials science. The course at } Lehigh University for this year was just held, and waiting until next year } is not a good option. } } If you teach or know about such a course, please contact me offline and I } will it forward the information. } } Thanks very much, } } --John Chandler } Fort Collins, CO } jchandler-at-ial-fa.com } 970.217.1321 } } } ==============================Original Headers============================== } 5, 22 -- From jchandler-at-ial-fa.com Mon Jul 18 11:51:41 2005 } 5, 22 -- Received: from sole.serverhost.net (sole.serverhost.net [216.71.84.188]) } 5, 22 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6IGpfq9019267 } 5, 22 -- for {Microscopy-at-microscopy.com} ; Mon, 18 Jul 2005 11:51:41 -0500 } 5, 22 -- Received: from JohnChandler (proxy1.lsil.com [147.145.40.41]) } 5, 22 -- by sole.serverhost.net (8.11.6/8.11.6) with SMTP id j6IGpeg09884 } 5, 22 -- for {Microscopy-at-microscopy.com} ; Mon, 18 Jul 2005 12:51:40 -0400 } 5, 22 -- Message-ID: {004c01c58bb8$d6f9ddf0$02000000-at-JohnChandler} } 5, 22 -- Reply-To: "John Chandler" {jchandler-at-ial-fa.com} } 5, 22 -- From: "John Chandler" {jchandler-at-ial-fa.com} } 5, 22 -- To: {Microscopy-at-microscopy.com} } 5, 22 -- Subject: Training: looking for TEM short course } 5, 22 -- Date: Mon, 18 Jul 2005 10:50:43 -0600 } 5, 22 -- Organization: Insight Analytical Labs } 5, 22 -- MIME-Version: 1.0 } 5, 22 -- Content-Type: text/plain; } 5, 22 -- charset="iso-8859-1" } 5, 22 -- Content-Transfer-Encoding: 7bit } 5, 22 -- X-Priority: 3 } 5, 22 -- X-MSMail-Priority: Normal } 5, 22 -- X-Mailer: Microsoft Outlook Express 6.00.2800.1506 } 5, 22 -- x-mimeole: Produced By Microsoft MimeOLE V6.00.2800.1506 } ==============================End of - Headers============================== } }
==============================Original Headers============================== 8, 25 -- From JOHN.WHEATLEY-at-asu.edu Mon Jul 18 14:59:32 2005 8, 25 -- Received: from post5.inre.asu.edu (post5.inre.asu.edu [129.219.110.120]) 8, 25 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6IJxWeZ012538 8, 25 -- for {microscopy-at-microscopy.com} ; Mon, 18 Jul 2005 14:59:32 -0500 8, 25 -- Received: from conversion.post5.inre.asu.edu by asu.edu (PMDF V6.1-1X6 #30769) 8, 25 -- id {0IJU00301A56P0-at-asu.edu} for microscopy-at-microscopy.com; Mon, 8, 25 -- 18 Jul 2005 12:58:18 -0700 (MST) 8, 25 -- Received: from EX1.asurite.ad.asu.edu (ex1.asurite.ad.asu.edu [129.219.10.211]) 8, 25 -- by asu.edu (PMDF V6.1-1X6 #30769) with ESMTP id {0IJU00LMFA56BR-at-asu.edu} for 8, 25 -- microscopy-at-microscopy.com; Mon, 18 Jul 2005 12:58:18 -0700 (MST) 8, 25 -- Date: Mon, 18 Jul 2005 12:58:18 -0700 8, 25 -- From: John Wheatley {JOHN.WHEATLEY-at-asu.edu} 8, 25 -- Subject: RE: [Microscopy] Training: looking for TEM short course 8, 25 -- To: microscopy-at-microscopy.com 8, 25 -- Message-id: {109BCD17C302C843A084EB9A5D45111D033E85E4-at-ex1.asurite.ad.asu.edu} 8, 25 -- MIME-version: 1.0 8, 25 -- X-MIMEOLE: Produced By Microsoft Exchange V6.0.6603.0 8, 25 -- Content-type: text/plain; charset=us-ascii 8, 25 -- Thread-Topic: [Microscopy] Training: looking for TEM short course 8, 25 -- Thread-Index: AcWLuUERqRrk+3XPQ/ertLOOe6eSagANYuxg 8, 25 -- Content-class: urn:content-classes:message 8, 25 -- X-MS-Has-Attach: 8, 25 -- X-MS-TNEF-Correlator: 8, 25 -- Content-Transfer-Encoding: 8bit 8, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id j6IJxWeZ012538 ==============================End of - Headers==============================
Thanks so much for getting back to me on your training course. I had looked at your website before polling the microscopy listserv, and it seemed to have lots of what is needed. ASU is one of the few centers for EM that I looked at before asking the question. I will pass along your information.
With best regards,
--John | jchandler-at-ial-fa.com | 970.217.1321
----- Original Message ----- X-from: {JOHN.WHEATLEY-at-asu.edu} To: {jchandler-at-ial-fa.com} Sent: Monday, July 18, 2005 1:59 PM
Dear Listers,
Below is a compilation of the replies I received to my recent question about Cu vs. Ni grids for immunolabeling. I usually don't do this without express permission, but most everybody posted to the list anyway, and I made sure to edit those that didn't. If it bothers anyone, let me know and I will never do it again.
In summary, it appears that copper can and will react with salts in buffers and other solutions, although some people still have decent luck. This problem can be ameliorated by using coated Cu grids and only wetting the coated side and by shortening incubation times
Also, Marc Pypaert had the interesting suggestion of just using Ni grids for everything, since they're not much more expensive, thereby avoiding the problem entirely. Any thoughts on this?
Many thanks to everyone who replied! I now have a much better response to our clients' questions on this matter (not to mention my own).
Cheers, Randy
Over the years I have frequently seen cases where the samples themselves have reacted with the copper grid. Two examples:
1) Looking at Fe-S precipitates in magnetotactic bacteria, when the precipitates were ugly and analyzed as copper sulphide. Our surmise was that the iron and copper had undergone ion exchange while the grid (and carbon film) was wet with the culture medium. When we used nickel grids, the precipitates were well formed and composed of iron and sulphur.
2) Looking at Si precipitates in Al-Si electronic bond wire. The specimens were prepared by embedding and microtoming, floating in DI water. The Si precipitates were surrounded by an ugly mess containing masses of copper. Again, presumed to me electrochemical reaction between the Al and the Cu grid, through the DI water medium, somehow preferentially at the Si precipitates. Again, use of Ni grids produced good pictures allowing us to characterize the precipitation.
Having said all that, we continue to default to use copper grids!
Anthony J. Garratt-Reed, M.A., D.Phil. MIT Room 13-1027 77 Massachusetts Avenue Cambridge, MA 02139-4307 USA
-------------------------------------------
The main problem encountered with copper grids used for immunocytochemistry or immunogold labeling is the reaction of the copper with salts in the buffers used during labeling. This is a time-related reaction, and can usually be avoided by having short labeling runs and using grids with films on them. Gilder grids, available from major microscope supply vendors, are gold-coated copper grids, and are not that much more expensive than regular copper grids. These grids may be the variety used by researchers who have reported having no problems with their grids during their immuno runs. I usually use nickel grids for my work, but have used formvar-coated copper grids without problems for 1 day immuno runs so long as the film is intact and as long as I don't get clumsy and end up sinking my grids in my solutions (if I do sink them, I do a quick distilled water rinse, blot the back of the grids with filter paper until almost dry, then re-float them where I left off). As others have shared, the nickel grids are much sturdier, not too expensive, and are not a problem to handle as long as you use antimagnetic forceps. With the nickel grids, remember to correct for astigmatism in the TEM by using a hole in your sample on the nickel grid. This will accommodate for any inherent residual magnetic field in the grid itself.
Edward Haller Lab Manager, Diagnostic Electron Microscopy Lab University of South Florida Pathology Department Tampa, FL 33612
We have had problems with Cu grids, even Formvar coated ones. I have precoated copper grids with Parlodion by dipping, blotting and drying before use, with or with out a formvar coating. THat method seems to protect the copper from reacting with PBS or whatever. This is not original to me, but I cannot recall where I read this (among many other things I can no longer recall). I think it might have been one of those smart Swiss guys.
Gregory W. Erdos, Ph.D. Assistant Director, Biotechnology Program Scientific Director, Electron Microscopy P.O. Box 118525 217 Carr Hall University of Florida Gainesville, FL 32611 -------------------------------------------------------------------
I had a bad experience with formvar-copper grids in immunogold experiment many years ago when incubating primary antibody overnight and the rest done on the following morning. I am using Nickel grids now -- no more problems.
Ann Fook Yang EM Unit/ Unite EM Agriculture and Agri-Food Canada/Agriculture et Agroalimentaire Canada 960 Carling Ave/960 Boul Carling Ottawa,Ontario/Ottawa, Ontario Canada K1A 0C6 ------------------------------------------------------------------------ ------------
recently i did a long ultracentrifuge run to load some 10S particles onto a formvar-copper grid. the grids reacted with the phosphate buffer over the 4 hour spin time. not a pretty picture - i had to go back to my collaborator and get fresh material so we could put it onto formvar-nickel grids. wonderful gentleman.
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 ----------------------------------------------------------------
We used Cu grids for many years. Normally we use formvar + carbon coated grids for ICC since we have a lot less loss of sections. We also tend to do long incubations ...usually overnight. This is time efficient and also lets us use highly diluted antibody so also minimize background due to cross-reactions from contaminants in polyclonal antibodies.
Occasionally we would have a reaction due to copper oxidizing with resultant green solution. I attributed this to salts or traces of Tween 20 in the buffer and breaks in the coating exposing the Cu. As far as I can see, this is the only reason for not using Cu grids if you use coated grids. We have switched for the most part to using coated Ni grids to avoid this problem.
On the other hand, occasionally we need to do a double labeling using uncoated grids and both sides of the sections. I would hesitate to use Cu for this and prefer Ni. Gold would work but is both more expensive and more delicate than Ni grids.
Debby Sherman, Manager Life Science Microscopy Facility Purdue University S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 ------------------------------------------------------------------------ -
X-from my experience, coated copper grids will be fine for most (if not all) immunolabelling, providing the solutions do not wet the uncoated side of the grid. Uncoated copper grids always have been fine when I've used PBS.
However, copper grids (uncoated - or the uncoated side) usually will react with Tris-HCl buffers and some of the high salt buffer formulations that I've used. Longer incubation times (eg. overnight) cause more reaction with the grid than shorter times (eg. 60mins) - so you may be able to use copper grids without problems if incubations are short, even with buffer formulations that might react with the grid if left for longer periods. It also is more difficult to prevent wetting of the reverse (uncoated) side of coated copper grids if incubation times are long, and particularly if wetting agents are used in the buffer formulation.
When I started immunolabelling in the early 1980s - and hadn't yet heard about the use of nickel grids - I used uncoated copper grids (and PBS) for years without problems. My current lab generally uses a high salt Tris-HCl buffer which includes a small amount of Tween 20, so we routinely use nickel grids for immunolabelling. Nickel grids do have some disadvantages (eg. charge), but we run a multi-user facility, often training students or other inexperienced users, we've found nickel the best option to overcome potential "grid-reaction" problems. However, nickel grids certainly aren't essential for successful immunolabelling - you just need to be aware of the possible problems.
Dr Deborah Stenzel Analytical Electron Microscopy Facility Queensland University of Technology GPO Box 2434 Brisbane 4001 Australia
------------------------------------------------
I know (from sad experience) that Tris buffers will etch Cu grids - the antibody droplets turn a lovely blue and there is gunk all over the sections. I use Ni grids, only because I prefer Tris buffers (no real reason why, I guess) for IEM and my current PI cringes at buying Au grids. I know one person who does all of her labelling in phosphate buffer and uses Cu grids with no precipitate problems.
As for the other reasons you Googled....I've heard them, but never seen any documentation nor had personal experience.
Can't wait to hear if anyone has actual evidence for some of the other no-Cu reasons!
Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 ----------------------------------------------------------------
Hi Randy, I had problems with Cu grids when incubating sections for long periods - overnight for instance. The copper would react with whatever was available and form green-blue salts. Not unexpected. We switched to nickel grids for a while but they charge, gold but they are delicate...by this time we had shortened the incubation times used, and tried copper again... And had no problem using coated grids and incubation times of an hour or so. Grids are floated on drops of media so that only the coated side is wetted - I don't know if that is important.
Dr SJ Stowe Facility Coordinator ANU Electron Microscopy Unit ANU CRICOS#00120C
I only ran into the "use only Ni or Au grids" rule here, in my current job, and my predecessor certainly produced some superb images showing gold labelling of TEM sections. Blissfully unaware of this rule, I had been using copper grids for all EM immunolabelling. To get good labelling of one particular structure, which is about 40 nm diameter, I used uncoated thin-bar Cu grids so I could get labelling on both sides of the section - seemed to work just fine. I'll be interested to see the responses to your question.
Dr. Rosemary White Microscopy Centre CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 Australia
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
==============================Original Headers============================== 54, 25 -- From TindallR-at-missouri.edu Mon Jul 18 16:21:32 2005 54, 25 -- Received: from um-exproto8.um.umsystem.edu (um-exproto8.um.umsystem.edu [207.160.151.48]) 54, 25 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6ILLW7R028963 54, 25 -- for {microscopy-at-microscopy.com} ; Mon, 18 Jul 2005 16:21:32 -0500 54, 25 -- Received: from UM-EMAIL09.um.umsystem.edu ([207.160.151.31]) by um-exproto8.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 54, 25 -- Mon, 18 Jul 2005 16:21:31 -0500 54, 25 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 54, 25 -- Content-class: urn:content-classes:message 54, 25 -- MIME-Version: 1.0 54, 25 -- Content-Type: text/plain; 54, 25 -- charset="us-ascii" 54, 25 -- Subject: TEM: Immunocytochemistry and Cu grids 54, 25 -- Date: Mon, 18 Jul 2005 16:21:30 -0500 54, 25 -- Message-ID: {BA876152E8653240BE8572E897083EE79802FD-at-UM-EMAIL09.um.umsystem.edu} 54, 25 -- X-MS-Has-Attach: 54, 25 -- X-MS-TNEF-Correlator: 54, 25 -- Thread-Topic: TEM: Immunocytochemistry and Cu grids 54, 25 -- Thread-Index: AcWL3qpvaFt7UpRlRY2LVYYS/0TUZg== 54, 25 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 54, 25 -- To: {microscopy-at-microscopy.com} 54, 25 -- Cc: "Jensen, Cheryl A." {JensenC-at-missouri.edu} , 54, 25 -- "Chance, Deborah L." {ChanceD-at-health.missouri.edu} 54, 25 -- X-OriginalArrivalTime: 18 Jul 2005 21:21:31.0576 (UTC) FILETIME=[AAE06380:01C58BDE] 54, 25 -- Content-Transfer-Encoding: 8bit 54, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id j6ILLW7R028963 ==============================End of - Headers==============================
Dear Roger I am not sure is tungsten carbide is similar to SiC or not. My experience with tungsten carbide basically was negative - this knife starts scratching the surface after just 50+ 0.5 um sections of standard plastic embedded tissue (no hard inclusions etc). As manufacturer explained to me it's because of polycrystalline nature of the material - small crystals just became loose and left the edge creating ruff surface. From this point of view, amorphous (like glass) or monocrystal (like diamond) material is preferable for good sections. Sapphire (hardness is 9) knifes were on the market for while without great success. From economical point of view, tungsten carbide knifes were also not so good - $100 each with ability to produce only 50+ good sections. If SiC acts in the similar way, then it would be difficult to use it in EM applications. Another aspect of using exotic materials - is it hydrophilic or hydrophobic? Could you use water with them? How easy to clean it up and handle? Sergey
At 07:00 AM 7/18/2005, you wrote:
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==============================Original Headers============================== 6, 25 -- From sryazant-at-ucla.edu Mon Jul 18 17:44:31 2005 6, 25 -- Received: from smtp-9.smtp.ucla.edu (smtp-9.smtp.ucla.edu [169.232.48.137]) 6, 25 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6IMiV81005437 6, 25 -- for {microscopy-at-microscopy.com} ; Mon, 18 Jul 2005 17:44:31 -0500 6, 25 -- Received: from mail.ucla.edu (mail.ucla.edu [169.232.47.141]) 6, 25 -- by smtp-9.smtp.ucla.edu (8.13.4/8.13.4) with ESMTP id j6IMiTCN030258 6, 25 -- for {microscopy-at-microscopy.com} ; Mon, 18 Jul 2005 15:44:29 -0700 6, 25 -- Received: from kopoba.ucla.edu (ts17-67.dialup.bol.ucla.edu [169.232.230.143]) 6, 25 -- (authenticated bits=0) 6, 25 -- by mail.ucla.edu (8.13.4/8.13.4) with ESMTP id j6IMiHs9001337 6, 25 -- (version=TLSv1/SSLv3 cipher=DES-CBC3-SHA bits=168 verify=NOT) 6, 25 -- for {microscopy-at-microscopy.com} ; Mon, 18 Jul 2005 15:44:21 -0700 6, 25 -- Message-Id: {6.1.2.0.2.20050718152757.032ce900-at-mail.ucla.edu} 6, 25 -- X-Sender: sryazant-at-mail.ucla.edu 6, 25 -- X-Mailer: QUALCOMM Windows Eudora Version 6.1.2.0 6, 25 -- Date: Mon, 18 Jul 2005 15:44:06 -0700 6, 25 -- To: microscopy-at-microscopy.com 6, 25 -- From: Sergey {sryazant-at-ucla.edu} 6, 25 -- Subject: Re: [Microscopy] Re: Alternatives to diamond knives? 6, 25 -- In-Reply-To: {200507181400.j6IE0bdx012784-at-ns.microscopy.com} 6, 25 -- References: {200507181400.j6IE0bdx012784-at-ns.microscopy.com} 6, 25 -- Mime-Version: 1.0 6, 25 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 6, 25 -- X-Probable-Spam: no 6, 25 -- X-Scanned-By: smtp.ucla.edu on 169.232.48.137 ==============================End of - Headers==============================
On Jul 18, 2005, at 5:48 PM, sryazant-at-ucla.edu wrote: } } Dear Roger } I am not sure is tungsten carbide is similar to SiC or not. My } experience } with tungsten carbide basically was negative - this knife starts } scratching } the surface after just 50+ 0.5 um sections of standard plastic } embedded } tissue (no hard inclusions etc). As manufacturer explained to me it's } because of polycrystalline nature of the material - small crystals } just } became loose and left the edge creating ruff surface. From this } point of } view, amorphous (like glass) or monocrystal (like diamond) material is } preferable for good sections. Sapphire (hardness is 9) knifes were } on the } market for while without great success. From economical point of } view, } tungsten carbide knifes were also not so good - $100 each with } ability to } produce only 50+ good sections. If SiC acts in the similar way, } then it } would be difficult to use it in EM applications. Another aspect of } using } exotic materials - is it hydrophilic or hydrophobic? Could you use } water } with them? How easy to clean it up and handle? Sergey
Thanks everyone, I've gotten lots of helpful advice and comments, so I'll try to sum up my conclusions.
Some background: Silicon carbide (SiC) is produced by a very old company Washington Mills located near Niagara Falls but production is in Illinois. Blessed folks; they sent me a sample of crystals for free.
It is made in tonnage quantities as clusters of glossy flat hexagonal black crystals up to a few centimeters across, and which are actually deep transparent blue and are crushed for abrasives and metallurgy. The cost of this industrial crystalline material is negligible. The Cree Corporation in NC makes also clear white SiC crystals and wafers for a fairly high price, but only sells them already faceted into clear diamond substitutes for jewelry. Such clear SiC crystals are termed "moissenite".
SiC crystals are very hard and brittle and the edge breaks when you try to sharpen them into knives using anything resembling normal gem faceting methods. But knives can be made using the proper lapping technique and the appropriate grades of diamond powder. Here is an interesting link on modern diamond powder technology:
At any rate, besides glass, only hard single crystals of diamond, sapphire, and silicon carbide seem to be appropriate for making ultramicrotome knives, which by their nature demand a perfectly homogeneous material. Sapphire knives were sold at one time by Diatome but the price was reputedly about $500 apiece and they became dull, unlike diamond knives, so the economics was questionable.
My SiC knives become dull after a few hundred slices, probably depending on the the hardness of the embedding plastic (glycol methacrylate works well), but the raw materials are inexpensive and my lapping machine only cost about $50 to build, so the labor of making them is inherently the major cost.
Since SiC crystals are difficult to sharpen without breaking the edge, I assume the exact same lapping techniques would also be appropriate for sapphire, which is softer tougher and less brittle than SiC; the latter are the very devil to sharpen without breaking. I learned how to make them through sheer stubbornness in order to make many serial sections with my Porter Blum MT-2 without paying $1000+ for a diamond knife.
I can only guess at the durability of sapphire knives, but the fact that they were once offered indicates that they might still be a viable alternative for some applications if they were only available at a lower cost.
My SiC crystals are epoxyed in a slot cut in the apex of a steel holder the same size and shape as a right angle glass knife. The clean crystals are hydrophilic (I assume the exposed surface layer at the atomic level is silicon oxide). I clean them by wiping the edge sideways with Teflon and don't have many wetting problems.
My conclusion is that I don't much want to go the trouble of trying to make money patenting, licensing and selling knives, but that I should publish my technique of making them, most likely in the Review of Scientific Instruments, and see if the world pays attention. I don't think the method of sharpening hard crystals to give very sharp edges has ever been published; I can't find it in the scientific literature. I seriously doubt it would work for diamonds without modification.
The only possible drawback to open publication is that no one maker could make a lot of money making them, and thus might not offer them, assuming they prove to be a modestly useful alternative to diamonds.
I assume that maybe the Chinese could sell disposable knives for $20 or so and they might catch on for student use and low budget and occasional use. Glass knives are clearly sharper for a few dozen sections and diamond knives clearly last longer.
-- Roger Baker Jr. 1303 Bentwood, Austin Tx, 78722
==============================Original Headers============================== 29, 19 -- From rcbaker-at-eden.infohwy.com Mon Jul 18 21:18:00 2005 29, 19 -- Received: from mx3.lsn.net (mx3.lsn.net [66.90.130.75]) 29, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6J2Hw75015970 29, 19 -- for {microscopy-at-microscopy.com} ; Mon, 18 Jul 2005 21:17:59 -0500 29, 19 -- Received: from [192.168.1.100] (66-90-146-134.dyn.grandenetworks.net [66.90.146.134]) 29, 19 -- by mx3.lsn.net (8.13.0.Beta3/8.12.8) with ESMTP id j6J2HkKD013027 29, 19 -- for {microscopy-at-microscopy.com} ; Mon, 18 Jul 2005 21:17:50 -0500 29, 19 -- Mime-Version: 1.0 (Apple Message framework v733) 29, 19 -- In-Reply-To: {200507182248.j6IMmje4011361-at-ns.microscopy.com} 29, 19 -- References: {200507182248.j6IMmje4011361-at-ns.microscopy.com} 29, 19 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 29, 19 -- Message-Id: {0FA04694-AC3E-4D7F-894D-13AD154CAF36-at-eden.infohwy.com} 29, 19 -- Content-Transfer-Encoding: 7bit 29, 19 -- From: Roger Baker {rcbaker-at-eden.infohwy.com} 29, 19 -- Subject: Re: [Microscopy] Alternatives to diamond knives? 29, 19 -- Date: Mon, 18 Jul 2005 21:17:44 -0500 29, 19 -- To: microscopy-at-microscopy.com 29, 19 -- X-Mailer: Apple Mail (2.733) 29, 19 -- X-Antivirus: Scanned by Vexira Antivirus 1.0.6 ==============================End of - Headers==============================
You should be aware that recently, gem-quality silicon carbide has been under production and is being marketed under its mineralogical name as "synthetic moissanite". This grade is significantly more free of defects and might yield a much more durable edge. I don't know the economics of producing a knife edge from this stock but I suspect that the material price is not prohibitively high, given the prices of stones that are being sold as jewelry and the usual mark-up in that field.
Sincerely,
John Twilley
rcbaker-at-eden.infohwy.com wrote:
} Thanks everyone, I've gotten lots of helpful advice and comments, so } I'll try to sum up my conclusions. } } Some background: Silicon carbide (SiC) is produced by a very old } company Washington Mills located near Niagara Falls but production is } in Illinois. Blessed folks; they sent me a sample of crystals for free. } } {http://www.medibix.com/company.jsp?company_id=10001791} } } It is made in tonnage quantities as clusters of glossy flat hexagonal } black crystals up to a few centimeters across, and which are actually } deep transparent blue and are crushed for abrasives and metallurgy. } The cost of this industrial crystalline material is negligible. The } Cree Corporation in NC makes also clear white SiC crystals and wafers } for a fairly high price, but only sells them already faceted into } clear diamond substitutes for jewelry. Such clear SiC crystals are } termed "moissenite". } } SiC crystals are very hard and brittle and the edge breaks when you } try to sharpen them into knives using anything resembling normal gem } faceting methods. But knives can be made using the proper lapping } technique and the appropriate grades of diamond powder. Here is an } interesting link on modern diamond powder technology: } } {http://www.ceramicindustry.com/CDA/ArticleInformation/features/ } BNP__Features__Item/0,2710,152388,00.html} } } At any rate, besides glass, only hard single crystals of diamond, } sapphire, and silicon carbide seem to be appropriate for making } ultramicrotome knives, which by their nature demand a perfectly } homogeneous material. Sapphire knives were sold at one time by } Diatome but the price was reputedly about $500 apiece and they became } dull, unlike diamond knives, so the economics was questionable. } } My SiC knives become dull after a few hundred slices, probably } depending on the the hardness of the embedding plastic (glycol } methacrylate works well), but the raw materials are inexpensive and } my lapping machine only cost about $50 to build, so the labor of } making them is inherently the major cost. } } Since SiC crystals are difficult to sharpen without breaking the } edge, I assume the exact same lapping techniques would also be } appropriate for sapphire, which is softer tougher and less brittle } than SiC; the latter are the very devil to sharpen without breaking. } I learned how to make them through sheer stubbornness in order to } make many serial sections with my Porter Blum MT-2 without paying } $1000+ for a diamond knife. } } I can only guess at the durability of sapphire knives, but the fact } that they were once offered indicates that they might still be a } viable alternative for some applications if they were only available } at a lower cost. } } My SiC crystals are epoxyed in a slot cut in the apex of a steel } holder the same size and shape as a right angle glass knife. The } clean crystals are hydrophilic (I assume the exposed surface layer at } the atomic level is silicon oxide). I clean them by wiping the edge } sideways with Teflon and don't have many wetting problems. } } My conclusion is that I don't much want to go the trouble of trying } to make money patenting, licensing and selling knives, but that I } should publish my technique of making them, most likely in the Review } of Scientific Instruments, and see if the world pays attention. I } don't think the method of sharpening hard crystals to give very sharp } edges has ever been published; I can't find it in the scientific } literature. I seriously doubt it would work for diamonds without } modification. } } The only possible drawback to open publication is that no one maker } could make a lot of money making them, and thus might not offer them, } assuming they prove to be a modestly useful alternative to diamonds. } } I assume that maybe the Chinese could sell disposable knives for $20 } or so and they might catch on for student use and low budget and } occasional use. Glass knives are clearly sharper for a few dozen } sections and diamond knives clearly last longer. } } -- Roger Baker Jr. } 1303 Bentwood, } Austin Tx, 78722 } } } ==============================Original Headers============================== } 29, 19 -- From rcbaker-at-eden.infohwy.com Mon Jul 18 21:18:00 2005 } 29, 19 -- Received: from mx3.lsn.net (mx3.lsn.net [66.90.130.75]) } 29, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6J2Hw75015970 } 29, 19 -- for {microscopy-at-microscopy.com} ; Mon, 18 Jul 2005 21:17:59 -0500 } 29, 19 -- Received: from [192.168.1.100] (66-90-146-134.dyn.grandenetworks.net [66.90.146.134]) } 29, 19 -- by mx3.lsn.net (8.13.0.Beta3/8.12.8) with ESMTP id j6J2HkKD013027 } 29, 19 -- for {microscopy-at-microscopy.com} ; Mon, 18 Jul 2005 21:17:50 -0500 } 29, 19 -- Mime-Version: 1.0 (Apple Message framework v733) } 29, 19 -- In-Reply-To: {200507182248.j6IMmje4011361-at-ns.microscopy.com} } 29, 19 -- References: {200507182248.j6IMmje4011361-at-ns.microscopy.com} } 29, 19 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed } 29, 19 -- Message-Id: {0FA04694-AC3E-4D7F-894D-13AD154CAF36-at-eden.infohwy.com} } 29, 19 -- Content-Transfer-Encoding: 7bit } 29, 19 -- From: Roger Baker {rcbaker-at-eden.infohwy.com} } 29, 19 -- Subject: Re: [Microscopy] Alternatives to diamond knives? } 29, 19 -- Date: Mon, 18 Jul 2005 21:17:44 -0500 } 29, 19 -- To: microscopy-at-microscopy.com } 29, 19 -- X-Mailer: Apple Mail (2.733) } 29, 19 -- X-Antivirus: Scanned by Vexira Antivirus 1.0.6 } ==============================End of - Headers==============================
==============================Original Headers============================== 9, 18 -- From jtwilley-at-sprynet.com Mon Jul 18 22:24:32 2005 9, 18 -- Received: from pop-satin.atl.sa.earthlink.net (pop-satin.atl.sa.earthlink.net [207.69.195.63]) 9, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6J3OW7G024406 9, 18 -- for {microscopy-at-microscopy.com} ; Mon, 18 Jul 2005 22:24:32 -0500 9, 18 -- Received: from pool-68-161-123-16.ny325.east.verizon.net ([68.161.123.16] helo=sprynet.com) 9, 18 -- by pop-satin.atl.sa.earthlink.net with esmtp (Exim 3.36 #10) 9, 18 -- id 1Duiid-0002BG-00; Mon, 18 Jul 2005 23:24:31 -0400 9, 18 -- Message-ID: {42DC7B7B.2CBEC0B5-at-sprynet.com} 9, 18 -- Date: Tue, 19 Jul 2005 00:03:07 -0400 9, 18 -- From: John Twilley {jtwilley-at-sprynet.com} 9, 18 -- X-Mailer: Mozilla 4.04 [en] (Win95; U) 9, 18 -- MIME-Version: 1.0 9, 18 -- To: microscopy-at-microscopy.com, rcbaker-at-eden.infohwy.com 9, 18 -- Subject: Re: [Microscopy] Re: Alternatives to diamond knives? 9, 18 -- References: {200507190218.j6J2IJo0016245-at-ns.microscopy.com} 9, 18 -- X-Corel-MessageType: EMail 9, 18 -- Content-Type: text/plain; charset=us-ascii 9, 18 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
I followed this thread by curiosity as I knew that copper is not always a good solution. And it was good to receive your message so that I know that there was no response that we missed. I think that the idea of giving a summary of the answers to the list, is a very respectful manner to contribute to its usefulness. Thanks a lot,
Sousan
TindallR-at-missouri.edu wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Dear Listers, } } Below is a compilation of the replies I received to my recent question } about Cu vs. Ni grids for immunolabeling. I usually don't do this } without express permission, but most everybody posted to the list } anyway, and I made sure to edit those that didn't. If it bothers } anyone, let me know and I will never do it again. } } In summary, it appears that copper can and will react with salts in } buffers and other solutions, although some people still have decent } luck. This problem can be ameliorated by using coated Cu grids and only } wetting the coated side and by shortening incubation times } } Also, Marc Pypaert had the interesting suggestion of just using Ni grids } for everything, since they're not much more expensive, thereby avoiding } the problem entirely. Any thoughts on this? } } Many thanks to everyone who replied! I now have a much better response } to our clients' questions on this matter (not to mention my own). } } Cheers, } Randy } } } Over the years I have frequently seen cases where the samples themselves } have reacted with the copper grid. Two examples: } } 1) Looking at Fe-S precipitates in magnetotactic bacteria, when the } precipitates were ugly and analyzed as copper sulphide. Our surmise was } that the iron and copper had undergone ion exchange while the grid (and } carbon film) was wet with the culture medium. When we used nickel } grids, the precipitates were well formed and composed of iron and } sulphur. } } 2) Looking at Si precipitates in Al-Si electronic bond wire. The } specimens were prepared by embedding and microtoming, floating in DI } water. The Si precipitates were surrounded by an ugly mess containing } masses of copper. Again, presumed to me electrochemical reaction } between the Al and the Cu grid, through the DI water medium, somehow } preferentially at the Si precipitates. Again, use of Ni grids produced } good pictures allowing us to characterize the precipitation. } } Having said all that, we continue to default to use copper grids! } } Anthony J. Garratt-Reed, M.A., D.Phil. } MIT Room 13-1027 } 77 Massachusetts Avenue } Cambridge, MA 02139-4307 } USA } } } ------------------------------------------- } } The main problem encountered with copper grids used for } immunocytochemistry or immunogold labeling is the reaction of the copper } with salts in the buffers used during labeling. This is a time-related } reaction, and can usually be avoided by having short labeling runs and } using grids with films on them. Gilder grids, available from major } microscope supply vendors, are gold-coated copper grids, and are not } that much more expensive than regular copper grids. These grids may be } the variety used by researchers who have reported having no problems } with their grids during their immuno runs. } I usually use nickel grids for my work, but have used } formvar-coated copper grids without problems for 1 day immuno runs so } long as the film is intact and as long as I don't get clumsy and end up } sinking my grids in my solutions (if I do sink them, I do a quick } distilled water rinse, blot the back of the grids with filter paper } until almost dry, then re-float them where I left off). As others have } shared, the nickel grids are much sturdier, not too expensive, and are } not a problem to handle as long as you use antimagnetic forceps. With } the nickel grids, remember to correct for astigmatism in the TEM by } using a hole in your sample on the nickel grid. This will accommodate } for any inherent residual magnetic field in the grid itself. } } Edward Haller } Lab Manager, Diagnostic Electron Microscopy Lab University of South } Florida Pathology Department Tampa, FL 33612 } } ------------------------------------------------------------ } } We have had problems with Cu grids, even Formvar coated ones. I have } precoated copper grids with Parlodion by dipping, blotting and drying } before use, with or with out a formvar coating. THat method seems to } protect the copper from reacting with PBS or whatever. This is not } original to me, but I cannot recall where I read this (among many other } things I can no longer recall). I think it might have been one of those } smart Swiss guys. } } Gregory W. Erdos, Ph.D. } Assistant Director, Biotechnology Program Scientific Director, Electron } Microscopy P.O. Box 118525 } 217 Carr Hall } University of Florida } Gainesville, FL 32611 } ------------------------------------------------------------------- } } I had a bad experience with formvar-copper grids in immunogold } experiment many years ago when incubating primary antibody overnight and } the rest done on the following morning. I am using Nickel grids now -- } no more problems. } } Ann Fook Yang } EM Unit/ Unite EM } Agriculture and Agri-Food Canada/Agriculture et Agroalimentaire Canada } 960 Carling Ave/960 Boul Carling } Ottawa,Ontario/Ottawa, Ontario } Canada } K1A 0C6 } ------------------------------------------------------------------------ } ------------ } } recently i did a long ultracentrifuge run to load some 10S particles } onto a formvar-copper grid. the grids reacted with the phosphate buffer } over the 4 hour spin time. not a pretty picture - i had to go back to } my collaborator and get fresh material so we could put it onto } formvar-nickel grids. wonderful gentleman. } } } Paul R. Hazelton, PhD } Electron Microscope Unit } University of Manitoba } Department of Medical Microbiology } 531 Basic Medical Sciences Building } 730 William Avenue } Winnipeg, Manitoba, Canada, R3E 0W3 } ---------------------------------------------------------------- } } We used Cu grids for many years. Normally we use formvar + carbon } coated grids for ICC since we have a lot less loss of sections. We also } tend to do long incubations ...usually overnight. This is time } efficient and also lets us use highly diluted antibody so also minimize } background due to cross-reactions from contaminants in polyclonal } antibodies. } } Occasionally we would have a reaction due to copper oxidizing with } resultant green solution. I attributed this to salts or traces of Tween } 20 in the buffer and breaks in the coating exposing the Cu. As far as I } can see, this is the only reason for not using Cu grids if you use } coated grids. We have switched for the most part to using coated Ni } grids to avoid this problem. } } On the other hand, occasionally we need to do a double labeling using } uncoated grids and both sides of the sections. I would hesitate to use } Cu for this and prefer Ni. Gold would work but is both more expensive } and more delicate than Ni grids. } } Debby Sherman, Manager } Life Science Microscopy Facility } Purdue University } S-052 Whistler Building } 170 S. University Street } West Lafayette, IN 47907 } ------------------------------------------------------------------------ } - } } X-from my experience, coated copper grids will be fine for most (if not } all) immunolabelling, providing the solutions do not wet the uncoated } side of the grid. Uncoated copper grids always have been fine when I've } used PBS. } } However, copper grids (uncoated - or the uncoated side) usually will } react with Tris-HCl buffers and some of the high salt buffer } formulations that I've used. Longer incubation times (eg. overnight) } cause more reaction with the grid than shorter times (eg. 60mins) - so } you may be able to use copper grids without problems if incubations are } short, even with buffer formulations that might react with the grid if } left for longer periods. It also is more difficult to prevent wetting } of the reverse (uncoated) side of coated copper grids if incubation } times are long, and particularly if wetting agents are used in the } buffer formulation. } } When I started immunolabelling in the early 1980s - and hadn't yet heard } about the use of nickel grids - I used uncoated copper grids (and PBS) } for years without problems. My current lab generally uses a high salt } Tris-HCl buffer which includes a small amount of Tween 20, so we } routinely use nickel grids for immunolabelling. Nickel grids do have } some disadvantages (eg. charge), but we run a multi-user facility, often } training students or other inexperienced users, we've found nickel the } best option to overcome potential "grid-reaction" problems. However, } nickel grids certainly aren't essential for successful immunolabelling - } you just need to be aware of the possible problems. } } Dr Deborah Stenzel } Analytical Electron Microscopy Facility } Queensland University of Technology } GPO Box 2434 } Brisbane 4001 } Australia } } ------------------------------------------------ } } I know (from sad experience) that Tris buffers will etch Cu grids - the } antibody droplets turn a lovely blue and there is gunk all over the } sections. I use Ni grids, only because I prefer Tris buffers (no real } reason why, I guess) for IEM and my current PI cringes at buying Au } grids. } I know one person who does all of her labelling in phosphate buffer and } uses Cu grids with no precipitate problems. } } As for the other reasons you Googled....I've heard them, but never seen } any documentation nor had personal experience. } } Can't wait to hear if anyone has actual evidence for some of the other } no-Cu reasons! } } Tamara Howard } Department of Cell Biology and Physiology } University of New Mexico - Health Sciences Center } Albuquerque, NM 87131 } ---------------------------------------------------------------- } } Hi Randy, } I had problems with Cu grids when incubating sections for long periods - } overnight for instance. The copper would react with whatever was } available and form green-blue salts. Not unexpected. We switched to } nickel grids for a while but they charge, gold but they are } delicate...by this time we had shortened the incubation times used, and } tried copper again... And had no problem using coated grids and } incubation times of an hour or so. Grids are floated on drops of media } so that only the coated side is wetted - I don't know if that is } important. } } Dr SJ Stowe } Facility Coordinator } ANU Electron Microscopy Unit } ANU CRICOS#00120C } } } I only ran into the "use only Ni or Au grids" rule here, in my current } job, and my predecessor certainly produced some superb images showing } gold labelling of TEM sections. Blissfully unaware of this rule, I had } been using copper grids for all EM immunolabelling. To get good } labelling of one particular structure, which is about 40 nm diameter, I } used uncoated thin-bar Cu grids so I could get labelling on both sides } of the section - seemed to work just fine. I'll be interested to see } the responses to your question. } } Dr. Rosemary White } Microscopy Centre } CSIRO Plant Industry } GPO Box 1600 } Canberra, ACT 2601 } Australia } } } } } } } } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility---We Do Small Well! } W122 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-2227 } Email: tindallr-at-missouri.edu } Web: http://www.emc.missouri.edu } } } } } } } } ==============================Original Headers============================== } 54, 25 -- From TindallR-at-missouri.edu Mon Jul 18 16:21:32 2005 } 54, 25 -- Received: from um-exproto8.um.umsystem.edu (um-exproto8.um.umsystem.edu [207.160.151.48]) } 54, 25 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6ILLW7R028963 } 54, 25 -- for {microscopy-at-microscopy.com} ; Mon, 18 Jul 2005 16:21:32 -0500 } 54, 25 -- Received: from UM-EMAIL09.um.umsystem.edu ([207.160.151.31]) by um-exproto8.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); } 54, 25 -- Mon, 18 Jul 2005 16:21:31 -0500 } 54, 25 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 } 54, 25 -- Content-class: urn:content-classes:message } 54, 25 -- MIME-Version: 1.0 } 54, 25 -- Content-Type: text/plain; } 54, 25 -- charset="us-ascii" } 54, 25 -- Subject: TEM: Immunocytochemistry and Cu grids } 54, 25 -- Date: Mon, 18 Jul 2005 16:21:30 -0500 } 54, 25 -- Message-ID: {BA876152E8653240BE8572E897083EE79802FD-at-UM-EMAIL09.um.umsystem.edu} } 54, 25 -- X-MS-Has-Attach: } 54, 25 -- X-MS-TNEF-Correlator: } 54, 25 -- Thread-Topic: TEM: Immunocytochemistry and Cu grids } 54, 25 -- Thread-Index: AcWL3qpvaFt7UpRlRY2LVYYS/0TUZg== } 54, 25 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} } 54, 25 -- To: {microscopy-at-microscopy.com} } 54, 25 -- Cc: "Jensen, Cheryl A." {JensenC-at-missouri.edu} , } 54, 25 -- "Chance, Deborah L." {ChanceD-at-health.missouri.edu} } 54, 25 -- X-OriginalArrivalTime: 18 Jul 2005 21:21:31.0576 (UTC) FILETIME=[AAE06380:01C58BDE] } 54, 25 -- Content-Transfer-Encoding: 8bit } 54, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id j6ILLW7R028963 } ==============================End of - Headers==============================
==============================Original Headers============================== 5, 20 -- From sousan.abolhassani-at-psi.ch Tue Jul 19 01:34:57 2005 5, 20 -- Received: from psi11.psi.ch (psi11.psi.ch [129.129.190.111]) 5, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id j6J6YuAl009619 5, 20 -- for {microscopy-at-microscopy.com} ; Tue, 19 Jul 2005 01:34:56 -0500 5, 20 -- Received: from 129.129.190.113 by psi11.psi.ch (InterScan E-Mail VirusWall NT); Tue, 19 Jul 2005 08:34:54 +0200 5, 20 -- Received: from [129.129.237.86] (PC3870.psi.ch [129.129.237.86]) by psi13.psi.ch with SMTP (Microsoft Exchange Internet Mail Service Version 5.5.2657.72) 5, 20 -- id JGAVKF5S; Tue, 19 Jul 2005 08:34:53 +0200 5, 20 -- Message-ID: {42DC9F0D.7050109-at-psi.ch} 5, 20 -- Date: Tue, 19 Jul 2005 08:34:53 +0200 5, 20 -- From: Sousan Abolhassani {sousan.abolhassani-at-psi.ch} 5, 20 -- Organization: Paul Scherrer Institut 5, 20 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.8) Gecko/20050511 5, 20 -- X-Accept-Language: en-us, en 5, 20 -- MIME-Version: 1.0 5, 20 -- To: microscopy-at-microscopy.com 5, 20 -- Subject: Re: [Microscopy] TEM: Immunocytochemistry and Cu grids 5, 20 -- References: {200507182124.j6ILOfrB002304-at-ns.microscopy.com} 5, 20 -- In-Reply-To: {200507182124.j6ILOfrB002304-at-ns.microscopy.com} 5, 20 -- Content-Type: text/plain; charset=us-ascii; format=flowed 5, 20 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
I need to prep some samples for SEM, but I'm not sure of the best way to go about it.
We have some bacteria that normally grow in a type of liquid nutrient medium as free, discrete and seperate motile cells. However, we have come across a strain that forms into 'mats' on the bottom of the culture vessel. It is these that we would like to examine by SEM.
The problem is that the 'mats' are not like a normal biofilm in that they are not strongly adherant to a solid surface, and the mat is not very robust. The mats will 'float' off the bottom of the culture vessel if they are disturbed and they will fragment into small flakes if the vessel is shaken or swirled. The fragments are usually a milimetre or so square.
Of course, the 'flakes' need to be flat in order to view them properly. I thought I might be able to pipette them, in a drop of the media they're already in, onto nucleopore membranes and allow them to settle, then try to remove the rest of the media and hope that they stick through the dehydration process... but i'm not sure they will. I'm planning on using HMDS.
I'm goig to try it just to see what happens... but has anyone prepared a sample like this that could suggest something that is know to work well?
Cheers,
-- Scott J. Coutts --------------------------------------------------------------- Bacterial Pathogenesis Research Group Department of Microbiology, Box 53, Monash University, 3800, Australia Phone: 9905 4838 Fax: 9905 4811 ---------------------------------------------------------------
==============================Original Headers============================== 8, 17 -- From scott.coutts-at-med.monash.edu.au Tue Jul 19 01:46:02 2005 8, 17 -- Received: from omta04sl.mx.bigpond.com (omta04sl.mx.bigpond.com [144.140.93.156]) 8, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6J6k1Km017389 8, 17 -- for {microscopy-at-microscopy.com} ; Tue, 19 Jul 2005 01:46:02 -0500 8, 17 -- Received: from [60.224.29.18] by omta04sl.mx.bigpond.com with ESMTP 8, 17 -- id {20050719064600.GCWB28893.omta04sl.mx.bigpond.com-at-[60.224.29.18]} 8, 17 -- for {microscopy-at-microscopy.com} ; Tue, 19 Jul 2005 06:46:00 +0000 8, 17 -- Message-ID: {42DCA1AB.5070904-at-med.monash.edu.au} 8, 17 -- Date: Tue, 19 Jul 2005 16:46:03 +1000 8, 17 -- From: Scott Coutts {scott.coutts-at-med.monash.edu.au} 8, 17 -- User-Agent: Mozilla Thunderbird 1.0.2 (Windows/20050317) 8, 17 -- X-Accept-Language: en-us, en 8, 17 -- MIME-Version: 1.0 8, 17 -- To: microscopy-at-microscopy.com 8, 17 -- Subject: Another SEM sample prep question 8, 17 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 8, 17 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Hi Scott, Is there a specific reason for looking at these bacteria with SEM? cheers, Roseamry
Dr. Rosemary White rosemary.white-at-csiro.au Microscopy Centre ph. 61-2-6246 5475 CSIRO Plant Industry mob. 61-0402 835 973 GPO Box 1600 fax. 61-2-6246 5334 Canberra, ACT 2601 Australia
} From: scott.coutts-at-med.monash.edu.au } Reply-To: microscopy-at-microscopy.com } Date: Tue, 19 Jul 2005 01:47:57 -0500 } To: rosemary.white-at-csiro.au } Subject: [Microscopy] Another SEM sample prep question } } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Hi All, } } I need to prep some samples for SEM, but I'm not sure of the best way to } go about it. } } We have some bacteria that normally grow in a type of liquid nutrient } medium as free, discrete and seperate motile cells. However, we have } come across a strain that forms into 'mats' on the bottom of the culture } vessel. It is these that we would like to examine by SEM. } } The problem is that the 'mats' are not like a normal biofilm in that } they are not strongly adherant to a solid surface, and the mat is not } very robust. The mats will 'float' off the bottom of the culture vessel } if they are disturbed and they will fragment into small flakes if the } vessel is shaken or swirled. The fragments are usually a milimetre or so } square. } } Of course, the 'flakes' need to be flat in order to view them properly. } I thought I might be able to pipette them, in a drop of the media } they're already in, onto nucleopore membranes and allow them to settle, } then try to remove the rest of the media and hope that they stick } through the dehydration process... but i'm not sure they will. I'm } planning on using HMDS. } } I'm goig to try it just to see what happens... but has anyone prepared a } sample like this that could suggest something that is know to work well? } } Cheers, } } -- } Scott J. Coutts } --------------------------------------------------------------- } Bacterial Pathogenesis Research Group } Department of Microbiology, } Box 53, Monash University, 3800, Australia } Phone: 9905 4838 Fax: 9905 4811 } --------------------------------------------------------------- } } ==============================Original Headers============================== } 8, 17 -- From scott.coutts-at-med.monash.edu.au Tue Jul 19 01:46:02 2005 } 8, 17 -- Received: from omta04sl.mx.bigpond.com (omta04sl.mx.bigpond.com } [144.140.93.156]) } 8, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6J6k1Km017389 } 8, 17 -- for {microscopy-at-microscopy.com} ; Tue, 19 Jul 2005 01:46:02 -0500 } 8, 17 -- Received: from [60.224.29.18] by omta04sl.mx.bigpond.com with ESMTP } 8, 17 -- id } {20050719064600.GCWB28893.omta04sl.mx.bigpond.com-at-[60.224.29.18]} } 8, 17 -- for {microscopy-at-microscopy.com} ; Tue, 19 Jul 2005 06:46:00 } +0000 } 8, 17 -- Message-ID: {42DCA1AB.5070904-at-med.monash.edu.au} } 8, 17 -- Date: Tue, 19 Jul 2005 16:46:03 +1000 } 8, 17 -- From: Scott Coutts {scott.coutts-at-med.monash.edu.au} } 8, 17 -- User-Agent: Mozilla Thunderbird 1.0.2 (Windows/20050317) } 8, 17 -- X-Accept-Language: en-us, en } 8, 17 -- MIME-Version: 1.0 } 8, 17 -- To: microscopy-at-microscopy.com } 8, 17 -- Subject: Another SEM sample prep question } 8, 17 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed } 8, 17 -- Content-Transfer-Encoding: 7bit } ==============================End of - Headers============================== }
==============================Original Headers============================== 4, 23 -- From Rosemary.White-at-csiro.au Tue Jul 19 01:52:08 2005 4, 23 -- Received: from vic-ironport-ext-out2.csiro.au (vic-ironport-ext-out2.csiro.au [150.229.64.38]) 4, 23 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6J6q7M5025152 4, 23 -- for {microscopy-at-microscopy.com} ; Tue, 19 Jul 2005 01:52:07 -0500 4, 23 -- Received: from exgw1-mel.nexus.csiro.au (138.194.3.56) 4, 23 -- by vic-ironport-ext-out2.csiro.au with ESMTP; 19 Jul 2005 16:52:05 +1000 4, 23 -- X-BrightmailFiltered: true 4, 23 -- X-Brightmail-Tracker: AAAAAQAAA+k= 4, 23 -- X-IronPort-AV: i="3.93,298,1114956000"; 4, 23 -- d="scan'208"; a="46134978:sNHT22338420" 4, 23 -- Received: from [152.83.167.45] ([152.83.167.45]) by exgw1-mel.nexus.csiro.au with Microsoft SMTPSVC(5.0.2195.6713); 4, 23 -- Tue, 19 Jul 2005 16:52:04 +1000 4, 23 -- User-Agent: Microsoft-Entourage/10.1.4.030702.0 4, 23 -- Date: Tue, 19 Jul 2005 16:53:19 +1000 4, 23 -- Subject: Re: [Microscopy] Another SEM sample prep question 4, 23 -- From: Rosemary White {Rosemary.White-at-csiro.au} 4, 23 -- To: {microscopy-at-microscopy.com} 4, 23 -- Message-ID: {BF02E07F.10AE9%Rosemary.White-at-csiro.au} 4, 23 -- In-Reply-To: {200507190647.j6J6lvFi022584-at-ns.microscopy.com} 4, 23 -- Mime-version: 1.0 4, 23 -- Content-type: text/plain; charset="US-ASCII" 4, 23 -- Content-transfer-encoding: 7bit 4, 23 -- X-OriginalArrivalTime: 19 Jul 2005 06:52:04.0345 (UTC) FILETIME=[5F326690:01C58C2E] ==============================End of - Headers==============================
IMAQUE IMAGING INC CALL 571-437-6593 ----- Original Message ----- X-from: {luce-at-earthtech.org} To: {JSMIT51-at-TAMPABAY.RR.COM} Sent: Sunday, July 17, 2005 7:05 PM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (K.venner-at-ion.ucl.ac.uk) from http://www.microscopy.com/MLFormMail.html on Tuesday, July 19, 2005 at 04:02:00 ---------------------------------------------------------------------------
Email: K.venner-at-ion.ucl.ac.uk Name: kerrie
Organization: Institute of Neurology, University College London UK
Title-Subject: [Filtered] MListserver: digital ccd cameras versus digital plate system for TEM
Question: Many thanks to everyone who took the time and kindly contributed to the very long debate my query sparked up. All contributions have been compiled and have enabled me to present a balanced working view of the systems currently available.
there is a very simple way to look at bacteria in the SEM. just puch teh bacteria/liquid through a .45 micron filter. then process the filter paper for the SEM. this is a tried and true method. john
--- scott.coutts-at-med.monash.edu.au wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Hi All, } } I need to prep some samples for SEM, but I'm not } sure of the best way to } go about it. } } We have some bacteria that normally grow in a type } of liquid nutrient } medium as free, discrete and seperate motile cells. } However, we have } come across a strain that forms into 'mats' on the } bottom of the culture } vessel. It is these that we would like to examine by } SEM. } } The problem is that the 'mats' are not like a normal } biofilm in that } they are not strongly adherant to a solid surface, } and the mat is not } very robust. The mats will 'float' off the bottom of } the culture vessel } if they are disturbed and they will fragment into } small flakes if the } vessel is shaken or swirled. The fragments are } usually a milimetre or so } square. } } Of course, the 'flakes' need to be flat in order to } view them properly. } I thought I might be able to pipette them, in a drop } of the media } they're already in, onto nucleopore membranes and } allow them to settle, } then try to remove the rest of the media and hope } that they stick } through the dehydration process... but i'm not sure } they will. I'm } planning on using HMDS. } } I'm goig to try it just to see what happens... but } has anyone prepared a } sample like this that could suggest something that } is know to work well? } } Cheers, } } -- } Scott J. Coutts } --------------------------------------------------------------- } Bacterial Pathogenesis Research Group } Department of Microbiology, } Box 53, Monash University, 3800, Australia } Phone: 9905 4838 Fax: 9905 4811 } --------------------------------------------------------------- } } ==============================Original } Headers============================== } 8, 17 -- From scott.coutts-at-med.monash.edu.au Tue Jul } 19 01:46:02 2005 } 8, 17 -- Received: from omta04sl.mx.bigpond.com } (omta04sl.mx.bigpond.com [144.140.93.156]) } 8, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with } ESMTP id j6J6k1Km017389 } 8, 17 -- for {microscopy-at-microscopy.com} ; Tue, 19 } Jul 2005 01:46:02 -0500 } 8, 17 -- Received: from [60.224.29.18] by } omta04sl.mx.bigpond.com with ESMTP } 8, 17 -- id } {20050719064600.GCWB28893.omta04sl.mx.bigpond.com-at-[60.224.29.18]} } 8, 17 -- for {microscopy-at-microscopy.com} ; } Tue, 19 Jul 2005 06:46:00 +0000 } 8, 17 -- Message-ID: } {42DCA1AB.5070904-at-med.monash.edu.au} } 8, 17 -- Date: Tue, 19 Jul 2005 16:46:03 +1000 } 8, 17 -- From: Scott Coutts } {scott.coutts-at-med.monash.edu.au} } 8, 17 -- User-Agent: Mozilla Thunderbird 1.0.2 } (Windows/20050317) } 8, 17 -- X-Accept-Language: en-us, en } 8, 17 -- MIME-Version: 1.0 } 8, 17 -- To: microscopy-at-microscopy.com } 8, 17 -- Subject: Another SEM sample prep question } 8, 17 -- Content-Type: text/plain; } charset=ISO-8859-1; format=flowed } 8, 17 -- Content-Transfer-Encoding: 7bit } ==============================End of - } Headers============================== }
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Interesting problem. The "matts" you refer to are likely to fragment or worse if you use the usual filter-based preparation methods . First question: do you have access to cryoSEM? If yes, this would be the best way to do your samples. Freeze in a high-pressure freezer, if available, or by plunging into slush nitrogen, then do the cryo SEM. This is the method most likely to give you the unaltered structure of the matts and the critters therein. If you don't have access to cryoSEM, then the best way is to use minifuge tubes or the like. Let the samples sit in the tube in fix, then the EtOH dehydration steps, being very careful when you withdraw the fluid to change it. Add the fluid for the next step as you withdraw the old fluid to minimize disturbance of the matts, and maybe add the fluid for each succeeding step down the side of the tube. I suggest a 2:1 1:1 1:2 EtOH:HMDS series before the 3 changes in 100% HMDS. I've found it helpful to deposit the last HMDS + sample drops on a sputter-coated membrane filter* stuck to a SEM stub for drying. This further minimizes handling. * 0.22 micron "nucleopore" type -- the ones with the nice, round holes, not the torturous-path type of filter. Sputter coat both sides of the filter before sticking to the stub, best, or stick to the stub with conductive carbon tabs, then sputter coat. Then drop on the samples in HMDS. (And yes, sputter coat for viewing in the SEM.)
Phil
} Hi All, } } I need to prep some samples for SEM, but I'm not sure of the best way to } go about it. } } We have some bacteria that normally grow in a type of liquid nutrient } medium as free, discrete and seperate motile cells. However, we have } come across a strain that forms into 'mats' on the bottom of the culture } vessel. It is these that we would like to examine by SEM. } } The problem is that the 'mats' are not like a normal biofilm in that } they are not strongly adherant to a solid surface, and the mat is not } very robust. The mats will 'float' off the bottom of the culture vessel } if they are disturbed and they will fragment into small flakes if the } vessel is shaken or swirled. The fragments are usually a milimetre or so } square. } } Of course, the 'flakes' need to be flat in order to view them properly. } I thought I might be able to pipette them, in a drop of the media } they're already in, onto nucleopore membranes and allow them to settle, } then try to remove the rest of the media and hope that they stick } through the dehydration process... but i'm not sure they will. I'm } planning on using HMDS. } } I'm goig to try it just to see what happens... but has anyone prepared a } sample like this that could suggest something that is know to work well? } } Cheers, } } -- } Scott J. Coutts } --------------------------------------------------------------- } Bacterial Pathogenesis Research Group } Department of Microbiology, } Box 53, Monash University, 3800, Australia } Phone: 9905 4838 Fax: 9905 4811 } --------------------------------------------------------------- } } ==============================Original Headers============================== } 8, 17 -- From scott.coutts-at-med.monash.edu.au Tue Jul 19 01:46:02 2005 } 8, 17 -- Received: from omta04sl.mx.bigpond.com } (omta04sl.mx.bigpond.com [144.140.93.156]) } 8, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id } j6J6k1Km017389 } 8, 17 -- for {microscopy-at-microscopy.com} ; Tue, 19 Jul 2005 } 01:46:02 -0500 } 8, 17 -- Received: from [60.224.29.18] by omta04sl.mx.bigpond.com with ESMTP } 8, 17 -- id } {20050719064600.GCWB28893.omta04sl.mx.bigpond.com-at-[60.224.29.18]} } 8, 17 -- for {microscopy-at-microscopy.com} ; Tue, 19 Jul 2005 } 06:46:00 +0000 } 8, 17 -- Message-ID: {42DCA1AB.5070904-at-med.monash.edu.au} } 8, 17 -- Date: Tue, 19 Jul 2005 16:46:03 +1000 } 8, 17 -- From: Scott Coutts {scott.coutts-at-med.monash.edu.au} } 8, 17 -- User-Agent: Mozilla Thunderbird 1.0.2 (Windows/20050317) } 8, 17 -- X-Accept-Language: en-us, en } 8, 17 -- MIME-Version: 1.0 } 8, 17 -- To: microscopy-at-microscopy.com } 8, 17 -- Subject: Another SEM sample prep question } 8, 17 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed } 8, 17 -- Content-Transfer-Encoding: 7bit } ==============================End of - Headers==============================
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) http://www.ansci.wisc.edu/microscopy.htm
==============================Original Headers============================== 5, 27 -- From peoshel-at-wisc.edu Tue Jul 19 09:11:41 2005 5, 27 -- Received: from smtp7.wiscmail.wisc.edu (hagen.doit.wisc.edu [144.92.197.163]) 5, 27 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6JEBeEu028672 5, 27 -- for {microscopy-at-microscopy.com} ; Tue, 19 Jul 2005 09:11:41 -0500 5, 27 -- Received: from avs-daemon.smtp7.wiscmail.wisc.edu by smtp7.wiscmail.wisc.edu 5, 27 -- (iPlanet Messaging Server 5.2 HotFix 2.04 (built Feb 8 2005)) 5, 27 -- id {0IJV00B1UORFO1-at-smtp7.wiscmail.wisc.edu} for microscopy-at-microscopy.com; 5, 27 -- Tue, 19 Jul 2005 09:11:39 -0500 (CDT) 5, 27 -- Received: from [10.25.102.29] (ansci.wisc.edu [144.92.132.175]) 5, 27 -- by smtp7.wiscmail.wisc.edu 5, 27 -- (iPlanet Messaging Server 5.2 HotFix 2.04 (built Feb 8 2005)) 5, 27 -- with ESMTPSA id {0IJV00MVUORDC9-at-smtp7.wiscmail.wisc.edu} for 5, 27 -- microscopy-at-microscopy.com; Tue, 19 Jul 2005 09:11:38 -0500 (CDT) 5, 27 -- Date: Tue, 19 Jul 2005 09:11:38 -0500 5, 27 -- From: Philip Oshel {peoshel-at-wisc.edu} 5, 27 -- Subject: Re: [Microscopy] Another SEM sample prep question 5, 27 -- In-reply-to: {200507190646.j6J6kc1d018856-at-ns.microscopy.com} 5, 27 -- X-Sender: peoshel-at-wiscmail.wisc.edu 5, 27 -- To: microscopy-at-microscopy.com 5, 27 -- Message-id: {p05210605bf02b64f46f3-at-[10.25.102.29]} 5, 27 -- MIME-version: 1.0 5, 27 -- Content-type: text/plain; format=flowed; charset=us-ascii 5, 27 -- Content-transfer-encoding: 7BIT 5, 27 -- X-Spam-Report: AuthenticatedSender=yes, SenderIP=144.92.132.175 5, 27 -- X-Spam-PmxInfo: Server=avs-7, Version=4.7.1.128075, Antispam-Engine: 2.0.3.1, 5, 27 -- Antispam-Data: 2005.7.19.11, SenderIP=144.92.132.175 5, 27 -- References: {200507190646.j6J6kc1d018856-at-ns.microscopy.com} ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (tantt-at-umich.edu) from http://microscopy.com/MicroscopyListserver/MLFormMail.html on Tuesday, July 19, 2005 at 09:03:46 ---------------------------------------------------------------------------
A few more details on what I wrote on this subject before. Lyson has been making inks and papers for photo quality ink jet printing for some time. Their new system is the "Daylight Darkroom". It is a dedicated 4 to 7 ink system (depending on your printer) for black and white printing only. I think the price includes software and inks but no printer, the system only works with certain printers. It has gotten very favorable reviews in fine art photo magazines (May/June 2005 issue of Photo Techniques, the review might be on the magazine's website (phototechmag.com) or on Lyson's). Lyson will send you some sample prints if you register at their website. I suspect that if someone sent Lyson a digital TEM or SEM image they might make a print of that as a sales tool. I'll bet they have no idea that there is a market for their products in the EM field. I only wish I did more than a few prints per year of parts of ground up cells. Lyson is not the only firm making inks, papers and printing systems for fine art B&W photography. Check out InkjetMall.com or MediaStreet.com. If you want to stick with an ink jet and the manufacturer's inks and papers the Canon i9900 is the Editor's Choice in the June 28 2005 issue of PC magazine. The same printer is also the first choice in Photo Techniques May/June 2005 issue. Someone on the list also liked this printer. If you like HP printers the PhotoSmart 8450 will take an HP 'photo gray' cartridge set for doing black and white prints. Disclaimer: I don't have any financial interest in any of the firms or products I mentioned.
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 **********************************************
==============================Original Headers============================== 6, 29 -- From mcauliff-at-umdnj.edu Tue Jul 19 09:32:30 2005 6, 29 -- Received: from mail01.umdnj.edu (zix01.UMDNJ.EDU [130.219.34.124]) 6, 29 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6JEWU4T011889 6, 29 -- for {Microscopy-at-msa.microscopy.com} ; Tue, 19 Jul 2005 09:32:30 -0500 6, 29 -- Received: from zix01.umdnj.edu (ZixVPM [127.0.0.1]) 6, 29 -- by Outbound.umdnj.edu (Proprietary) with ESMTP id A46DEEC0D0 6, 29 -- for {Microscopy-at-msa.microscopy.com} ; Tue, 19 Jul 2005 10:32:29 -0400 (EDT) 6, 29 -- Received: from polaris.umdnj.edu (polarisa.UMDNJ.EDU [130.219.34.131]) 6, 29 -- by mail01.umdnj.edu (Proprietary) with ESMTP id F0CF1EC0C6 6, 29 -- for {Microscopy-at-msa.microscopy.com} ; Tue, 19 Jul 2005 10:32:27 -0400 (EDT) 6, 29 -- Received: from conversion-daemon.Polaris.umdnj.edu by Polaris.umdnj.edu 6, 29 -- (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 2004)) 6, 29 -- id {0IJV00401P5RJY-at-Polaris.umdnj.edu} (original mail from mcauliff-at-umdnj.edu) 6, 29 -- for Microscopy-at-msa.microscopy.com; Tue, 19 Jul 2005 10:32:27 -0400 (EDT) 6, 29 -- Received: from [127.0.0.1] ([10.138.2.240]) 6, 29 -- by Polaris.umdnj.edu (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 6, 29 -- 2004)) with ESMTP id {0IJV00IVXPOJ8M-at-Polaris.umdnj.edu} ; Tue, 6, 29 -- 19 Jul 2005 10:31:31 -0400 (EDT) 6, 29 -- Date: Tue, 19 Jul 2005 10:31:34 -0400 6, 29 -- From: Geoff McAuliffe {mcauliff-at-umdnj.edu} 6, 29 -- Subject: dye sub, ink jet etc for prints 6, 29 -- To: Microscopy-at-msa.microscopy.com, "John J. Bozzola" {bozzola-at-siu.edu} 6, 29 -- Message-id: {42DD0EC6.90103-at-umdnj.edu} 6, 29 -- MIME-version: 1.0 6, 29 -- Content-type: text/plain; format=flowed; charset=ISO-8859-1 6, 29 -- Content-transfer-encoding: 7BIT 6, 29 -- X-Accept-Language: en-us, en 6, 29 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 6, 29 -- Gecko/20040804 Netscape/7.2 (ax) ==============================End of - Headers==============================
hoffpajo-at-yahoo.com wrote: } } there is a very simple way to look at bacteria in the } SEM. just puch teh bacteria/liquid through a .45 } micron filter. then process the filter paper for the } SEM. this is a tried and true method. } john }
Yes, but that doesnt help me to lay mats of bacteria onto a support - filtering them is what we normally do for the free-living cells, but if I filter the mats, firstly there's a good chance that they;ll get stuck in the syringe, and secondly, they won't come out flat on the filter. THey'll be all bunched up or folded.
-- Scott J. Coutts --------------------------------------------------------------- Bacterial Pathogenesis Research Group Department of Microbiology, Box 53, Monash University, 3800, Australia Phone: 9905 4838 Fax: 9905 4811 ---------------------------------------------------------------
==============================Original Headers============================== 3, 19 -- From scott.coutts-at-med.monash.edu.au Tue Jul 19 09:53:59 2005 3, 19 -- Received: from omta05ps.mx.bigpond.com (omta05ps.mx.bigpond.com [144.140.83.195]) 3, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6JErwbY019859 3, 19 -- for {microscopy-at-microscopy.com} ; Tue, 19 Jul 2005 09:53:59 -0500 3, 19 -- Received: from [60.224.29.18] by omta05ps.mx.bigpond.com with ESMTP 3, 19 -- id {20050719145357.UTTR21584.omta05ps.mx.bigpond.com-at-[60.224.29.18]} 3, 19 -- for {microscopy-at-microscopy.com} ; Tue, 19 Jul 2005 14:53:57 +0000 3, 19 -- Message-ID: {42DD1404.3020406-at-med.monash.edu.au} 3, 19 -- Date: Wed, 20 Jul 2005 00:53:56 +1000 3, 19 -- From: Scott Coutts {scott.coutts-at-med.monash.edu.au} 3, 19 -- User-Agent: Mozilla Thunderbird 1.0.2 (Windows/20050317) 3, 19 -- X-Accept-Language: en-us, en 3, 19 -- MIME-Version: 1.0 3, 19 -- To: microscopy-at-microscopy.com 3, 19 -- Subject: Re: [Microscopy] Re: Another SEM sample prep question 3, 19 -- References: {200507191344.j6JDi0Zi028149-at-ns.microscopy.com} 3, 19 -- In-Reply-To: {200507191344.j6JDi0Zi028149-at-ns.microscopy.com} 3, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 3, 19 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
This is interesting information. Would you be willing to write it up for MIcroscopy Today? With illustrations and all. We are interested in running such an article, and there would be room for the details and so forth that you can't put in an email.
Phil (also still not entirely retired from Tech Ed. at Microscopy Today)
} Thanks everyone, I've gotten lots of helpful advice and comments, so } I'll try to sum up my conclusions. } } Some background: Silicon carbide (SiC) is produced by a very old } company Washington Mills located near Niagara Falls but production is } in Illinois. Blessed folks; they sent me a sample of crystals for free. } } {http://www.medibix.com/company.jsp?company_id=10001791} } } It is made in tonnage quantities as clusters of glossy flat hexagonal } black crystals up to a few centimeters across, and which are actually } deep transparent blue and are crushed for abrasives and metallurgy. } The cost of this industrial crystalline material is negligible. The } Cree Corporation in NC makes also clear white SiC crystals and wafers } for a fairly high price, but only sells them already faceted into } clear diamond substitutes for jewelry. Such clear SiC crystals are } termed "moissenite". } } SiC crystals are very hard and brittle and the edge breaks when you } try to sharpen them into knives using anything resembling normal gem } faceting methods. But knives can be made using the proper lapping } technique and the appropriate grades of diamond powder. Here is an } interesting link on modern diamond powder technology: } } {http://www.ceramicindustry.com/CDA/ArticleInformation/features/ } BNP__Features__Item/0,2710,152388,00.html} } } At any rate, besides glass, only hard single crystals of diamond, } sapphire, and silicon carbide seem to be appropriate for making } ultramicrotome knives, which by their nature demand a perfectly } homogeneous material. Sapphire knives were sold at one time by } Diatome but the price was reputedly about $500 apiece and they became } dull, unlike diamond knives, so the economics was questionable. } } My SiC knives become dull after a few hundred slices, probably } depending on the the hardness of the embedding plastic (glycol } methacrylate works well), but the raw materials are inexpensive and } my lapping machine only cost about $50 to build, so the labor of } making them is inherently the major cost. } } Since SiC crystals are difficult to sharpen without breaking the } edge, I assume the exact same lapping techniques would also be } appropriate for sapphire, which is softer tougher and less brittle } than SiC; the latter are the very devil to sharpen without breaking. } I learned how to make them through sheer stubbornness in order to } make many serial sections with my Porter Blum MT-2 without paying } $1000+ for a diamond knife. } } I can only guess at the durability of sapphire knives, but the fact } that they were once offered indicates that they might still be a } viable alternative for some applications if they were only available } at a lower cost. } } My SiC crystals are epoxyed in a slot cut in the apex of a steel } holder the same size and shape as a right angle glass knife. The } clean crystals are hydrophilic (I assume the exposed surface layer at } the atomic level is silicon oxide). I clean them by wiping the edge } sideways with Teflon and don't have many wetting problems. } } My conclusion is that I don't much want to go the trouble of trying } to make money patenting, licensing and selling knives, but that I } should publish my technique of making them, most likely in the Review } of Scientific Instruments, and see if the world pays attention. I } don't think the method of sharpening hard crystals to give very sharp } edges has ever been published; I can't find it in the scientific } literature. I seriously doubt it would work for diamonds without } modification. } } The only possible drawback to open publication is that no one maker } could make a lot of money making them, and thus might not offer them, } assuming they prove to be a modestly useful alternative to diamonds. } } I assume that maybe the Chinese could sell disposable knives for $20 } or so and they might catch on for student use and low budget and } occasional use. Glass knives are clearly sharper for a few dozen } sections and diamond knives clearly last longer. } } -- Roger Baker Jr. } 1303 Bentwood, } Austin Tx, 78722 } } } } } } } } } } } } } ==============================Original Headers============================== } 29, 19 -- From rcbaker-at-eden.infohwy.com Mon Jul 18 21:18:00 2005 } 29, 19 -- Received: from mx3.lsn.net (mx3.lsn.net [66.90.130.75]) } 29, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id } j6J2Hw75015970 } 29, 19 -- for {microscopy-at-microscopy.com} ; Mon, 18 Jul 2005 } 21:17:59 -0500 } 29, 19 -- Received: from [192.168.1.100] } (66-90-146-134.dyn.grandenetworks.net [66.90.146.134]) } 29, 19 -- by mx3.lsn.net (8.13.0.Beta3/8.12.8) with ESMTP id } j6J2HkKD013027 } 29, 19 -- for {microscopy-at-microscopy.com} ; Mon, 18 Jul 2005 } 21:17:50 -0500 } 29, 19 -- Mime-Version: 1.0 (Apple Message framework v733) } 29, 19 -- In-Reply-To: {200507182248.j6IMmje4011361-at-ns.microscopy.com} } 29, 19 -- References: {200507182248.j6IMmje4011361-at-ns.microscopy.com} } 29, 19 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed } 29, 19 -- Message-Id: {0FA04694-AC3E-4D7F-894D-13AD154CAF36-at-eden.infohwy.com} } 29, 19 -- Content-Transfer-Encoding: 7bit } 29, 19 -- From: Roger Baker {rcbaker-at-eden.infohwy.com} } 29, 19 -- Subject: Re: [Microscopy] Alternatives to diamond knives? } 29, 19 -- Date: Mon, 18 Jul 2005 21:17:44 -0500 } 29, 19 -- To: microscopy-at-microscopy.com } 29, 19 -- X-Mailer: Apple Mail (2.733) } 29, 19 -- X-Antivirus: Scanned by Vexira Antivirus 1.0.6 } ==============================End of - Headers==============================
-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax) http://www.ansci.wisc.edu/microscopy.htm
==============================Original Headers============================== 5, 27 -- From peoshel-at-wisc.edu Tue Jul 19 10:19:15 2005 5, 27 -- Received: from smtp7.wiscmail.wisc.edu (hagen.doit.wisc.edu [144.92.197.163]) 5, 27 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id j6JFJFLi027873 5, 27 -- for {microscopy-at-microscopy.com} ; Tue, 19 Jul 2005 10:19:15 -0500 5, 27 -- Received: from avs-daemon.smtp7.wiscmail.wisc.edu by smtp7.wiscmail.wisc.edu 5, 27 -- (iPlanet Messaging Server 5.2 HotFix 2.04 (built Feb 8 2005)) 5, 27 -- id {0IJV00F0QRVGWH-at-smtp7.wiscmail.wisc.edu} for microscopy-at-microscopy.com; 5, 27 -- Tue, 19 Jul 2005 10:18:52 -0500 (CDT) 5, 27 -- Received: from [10.25.102.29] (ansci.wisc.edu [144.92.132.175]) 5, 27 -- by smtp7.wiscmail.wisc.edu 5, 27 -- (iPlanet Messaging Server 5.2 HotFix 2.04 (built Feb 8 2005)) 5, 27 -- with ESMTPSA id {0IJV00G1BRVB3H-at-smtp7.wiscmail.wisc.edu} for 5, 27 -- microscopy-at-microscopy.com; Tue, 19 Jul 2005 10:18:48 -0500 (CDT) 5, 27 -- Date: Tue, 19 Jul 2005 10:18:49 -0500 5, 27 -- From: Philip Oshel {peoshel-at-wisc.edu} 5, 27 -- Subject: [Microscopy] Re: Alternatives to diamond knives? 5, 27 -- In-reply-to: {200507190219.j6J2JWRe017424-at-ns.microscopy.com} 5, 27 -- X-Sender: peoshel-at-wiscmail.wisc.edu 5, 27 -- To: microscopy-at-microscopy.com 5, 27 -- Message-id: {p05210609bf02c9c1d87a-at-[10.25.102.29]} 5, 27 -- MIME-version: 1.0 5, 27 -- Content-type: text/plain; format=flowed; charset=us-ascii 5, 27 -- Content-transfer-encoding: 7BIT 5, 27 -- X-Spam-Report: AuthenticatedSender=yes, SenderIP=144.92.132.175 5, 27 -- X-Spam-PmxInfo: Server=avs-3, Version=4.7.1.128075, Antispam-Engine: 2.0.3.1, 5, 27 -- Antispam-Data: 2005.7.19.14, SenderIP=144.92.132.175 5, 27 -- References: {200507190219.j6J2JWRe017424-at-ns.microscopy.com} ==============================End of - Headers==============================
I second Sousan's reply. It saves downloading a lot of different emails! I also found the information quite helpful.
Peggy
Peggy Sherwood Lab Associate, Photopathology Wellman Center for Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
-----Original Message----- X-from: TindallR-at-missouri.edu [mailto:TindallR-at-missouri.edu] Sent: Monday, July 18, 2005 5:22 PM To: Sherwood, Margaret
Dear Listers,
Below is a compilation of the replies I received to my recent question about Cu vs. Ni grids for immunolabeling. I usually don't do this without express permission, but most everybody posted to the list anyway, and I made sure to edit those that didn't. If it bothers anyone, let me know and I will never do it again.
In summary, it appears that copper can and will react with salts in buffe