by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id FAA23779 for {microscopy-at-sparc5.microscopy.com} ; Sat, 1 Sep 2001 05:54:34 -0500 (CDT) Received: from Dmrelion-at-aol.com by imo-m02.mx.aol.com (mail_out_v31_r1.4.) id f.16c.2107f6 (4007) for {microscopy-at-msa.microscopy.com} ; Sat, 1 Sep 2001 06:51:29 -0400 (EDT) Message-ID: {16c.2107f6.28c217b1-at-aol.com}
I have followed this discussion with some interest. A similar situation exists in the field of mass spectroscopy.
At one time there were many mass spectrographs and mass spectrometers being used. Mass spectrographs were those instruments in which the detected ions were spread out along a focal plane and a significant portion of the mass range was collected simultaneously on a photographic plate. Mass spectrometers were those instruments in which the detected ions were moved across a detector slit (by varying magnetic field or ion accelerating potential) and collected/detected, one m/q at a time, by either a Faraday cup or electron multiplier after the slit, providing a readout on an electronic device (electrometer usually). Mass spectroscope or mass spectroscopy was used as the general term to include either type of instrument.
Mass spectrographs are rarely seen any more and even if they are used, the photographic plate would probably be replaced by some type of CCD detector providing electronic readout so the original clear distinction becomes blurred.
I'm sure that an analogous situation exists in optical spectroscopy.
Don Marshall
Don Marshall RELION Industries PO Box 12 Bedford, MA 01730
I have followed this discussion with some interest. A similar situation exists in the field of mass spectroscopy.
At one time there were many mass spectrographs and mass spectrometers being used. Mass spectrographs were those instruments in which the detected ions were spread out along a focal plane and a significant portion of the mass range was collected simultaneously on a photographic plate. Mass spectrometers were those instruments in which the detected ions were moved across a detector slit (by varying magnetic field or ion accelerating potential) and collected/detected, one m/q at a time, by either a Faraday cup or electron multiplier after the slit, providing a readout on an electronic device (electrometer usually). Mass spectroscope or mass spectroscopy was used as the general term to include either type of instrument.
Mass spectrographs are rarely seen any more and even if they are used, the photographic plate would probably be replaced by some type of CCD detector providing electronic readout so the original clear distinction becomes blurred.
I'm sure that an analogous situation exists in optical spectroscopy.
Don Marshall
Don Marshall RELION Industries PO Box 12 Bedford, MA 01730
You do not state the exact type of silica you are embedding. SO...
In general methacrylates, whether LR White or such, will shrink much more than an epoxy like one of the EPON CLONE formulations. If I recall 20-25% shrinkage is what you get with LR White and a lot of polymers. Epoxy formulas are more like 5%. This lower shrinkage is preferred by me so that I can argue to customers that I have not changed the microstructrure through severe shrinkage during the polymerization. LR White is faster (with an accelerator) curing, if you are in a hurry. The EPON clone stuff takes overnight curing in an oven at 60 to 70 C. It's no big deal because you just prepare the samples at the end of the day and start sectioning in the morning.
I have personnaly done precipitated silicas, arc silicas, and fumed silicas using Epon. Silica fume should be no problem. I do not vacuum embbed these materials. Ppt'd silicas have a surface area of about 154-250 meters squared per gram and the epon will wet almost every pore between the primary or ultimate particles without vacuum. Capillary forces are strong enough on our products and others to wet almost 100% of what is there. One of the components in my EPON kit foams under partial vacuum and that is another reason I don't use vacuum. You might have to use vacuum in your case. However, test each EPON kit component separately to see how much vacuum it can tolerate before foaming.
We have made various sizes of agglomerated spheres of these particles since the mid 70s as I recall. Anyway, they can be fragile but epon does not break up the microstructure. One can section throgh the 'micro-dust' on the surface of these spheres without loss of material.
As you can see, EPON works on a wide range of 'porous silica' samples. There must be a reason why EM people like it? It works on a wide range of materials, wets well, sticks fairly well to them, and sections well.
Eponate 12 releases from polyethelene molds, bottle caps, etc. quite well for 1-4 cycles of mold use with EPON, FYI.
Hope this helps.
Paul Beauregard Senior Research Associate PPG Industries Monroeville Technical Center Monroeville, PA 15146
Opinions given are my own and not those of PPG Industries.
} Dear all } } I have a sample of dried porous silica that I would like to embed and } thin section. Could anyone give me any advice on the best resin and } protocol to use? The idea of using a methacrylate resin has been thrown } about (the idea, not the resin - I can assure you), however I would be } very grateful for any advice regarding any of the resins available. If } this is a bit specific then reply off-line to the address below. } } Thanks } Pippa } ---------------------- } Pippa Hawes, EMU } School of Chemistry } University of Bristol } Cantocks Close } Bristol UK } BS8 1TS } Pippa.Hawes-at-bristol.ac.uk } } } }
Earl Weltmer and Brad Johnson wrote: ============================================================ } Hello, } I need to replace the SE scintillator on my SEM (JEOL 5900), and I was } considering using a YAP scintillator instead of the typical phosphorous } scintillator. Does anyone have any experience or suggestions on this? ++++Brad Johnson
About ten years ago I ran some tests for signal on several scintillator types: YAP, YAG, standard aluminum coated scintillator & "uncoated" scintillator. The result at that time were surprising.
Using a JEOL 35C SEM & running the same specimen current & PMT voltage with each scintillator the best signal was given by the original aluminum coated scintillator.
The second best was the "uncoated" scintillator & it was only 80% of the signal the standard aluminum coated scintillator. The YAP & YAG gave only 60% of the standard aluminum scintillator.
I was very disappointed as the YAP & YAG were significantly more than the standard scintillator. The conclusion that I came to was that the YAP & YAG would probably give more longevity & could justify their extra cost but they were not fit for high resolution. The YAP & YAG would probably be suitable for probe work.
Keep in mind this experiment was done about ten years ago. Perhaps the crystals have improved in recent years & the data may need a re-evaluation. I am sure that there is someone who would be willing to criticize my results (maybe in Northern Calif.?) but this the data that I received at the time. +++++ Earl Weltmer ============================================================ We have offered P-47, YAG and YAP scintillators for SEM applications for more then twenty years. Earl is correct that the P-47 performance, **but when first installed**, does probably give a superior performance. However , not all YAG and YAP crystals come out of the "same cookie cutter." From what we have been able to deduce, a high quality YAG or YAP single crystal scintillator will perform so close to that of the P-47 that one usually has difficulty detecting the difference. However, as everyone knows, the P-47 once installed, in terms of performance, goes only down. The only question is how fast it will go down. And that rate of deterioration of performance depends among other things on the type of work being done. For example, the higher x-ray fluxes associated with EDS (and especially WDS) tend to cause a more rapid deterioration.
Also, if Earl's experiment was done as described, and if a YAG was tested against P-47 but using the same PMT, then one would expect to find an inferior performance, because the PMT that is optimum for P47 is not the same one that would have been optimum for YAG. Now this might be less of a problem today than it would have been ten years ago, but if the PMT was not changed, then the right test (IMHO) was not the one being performed. To make the comparison, one would have had to have taken out the S11 style and replaced it with an S20 PMT. This point is further explained on our website URL http://www.2spi.com/catalog/scintill/spi-yag.html
When a YAG or YAP scintillator is installed, its performance level is constant and does not deteriorate. So while the comparison might suggest some superiority at the very beginning, in most instances that we have seen, or have been led to believe, it is not long that the SEM running on YAG or YAP in general, is operating at a higher level of performance. And the SEM is not subjected to the downtime associated with the need to change a P-47 powder scintillator from time to time.
For those operating at low KV in the BSE mode (and using YAG or YAP), and high magnifications, the image is clearly less noisy. A collection of ten different references from different laboratories around the world, which have been collected on URL http://www.2spi.com/catalog/scintill/sem-tem.html at least to some, pretty much document the superiority of the single crystal scintillators over the powder scintillators. I guess one could always try to do more studies, but anyone that is familiar with these publications finds their conclusions pretty persuasive.
We have generally recommended that the SPI single crystal scintillators offered many advantages over the more traditional P-47 scintillator. We have been under the impression that those who have made the switch have been pretty happy with their results, though we do hear from time to time of someone who would not agree with that statement. Of course, the performance is related to the nature of the doping of the crystal, and as I said, not all these crystals come out of the same cookie cutter. Not all YAG's and YAP's are created equal. So when making comparisons, and statements about their scintillators, it would be helpful for one to state the brand of their scintillator crystal. These crystals are certainly not a generic entity where all are the same irrespective of the source.
Disclaimer: SPI Supplies is a major supplier of scintillators for SEM applications including P-47's, YAGs and YAPs.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
} } I have a sample of dried porous silica that I would like to embed and } thin section. Could anyone give me any advice on the best resin and } protocol to use? The idea of using a methacrylate resin has been thrown } about (the idea, not the resin - I can assure you), however I would be } very grateful for any advice regarding any of the resins available. If } this is a bit specific then reply off-line to the address below. } Pippa -
You'll probably do fine with LR White, Hard grade. Its low viscosity is a big help, but if the sample floats anyway you may need vacuum. If you've never used that resin, be cautious when selecting the embedding mold; it will dissolve some plastics & it won't harden in flat molds unless air is excluded. Gelatin capsules work.
Caroline
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
I have followed this discussion with some interest. A similar situation exists in the field of mass spectroscopy.
At one time there were many mass spectrographs and mass spectrometers being used. Mass spectrographs were those instruments in which the detected ions were spread out along a focal plane and a significant portion of the mass range was collected simultaneously on a photographic plate. Mass spectrometers were those instruments in which the detected ions were moved across a detector slit (by varying magnetic field or ion accelerating potential) and collected/detected, one m/q at a time, by either a Faraday cup or electron multiplier after the slit, providing a readout on an electronic device (electrometer usually). Mass spectroscope or mass spectroscopy was used as the general term to include either type of instrument.
Mass spectrographs are rarely seen any more and even if they are used, the photographic plate would probably be replaced by some type of CCD detector providing electronic readout so the original clear distinction becomes blurred.
I'm sure that an analogous situation exists in optical spectroscopy.
Don Marshall
Don Marshall RELION Industries PO Box 12 Bedford, MA 01730
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id AAA25972 for dist-Microscopy; Sun, 2 Sep 2001 00:06:16 -0500 (CDT) Received: from no_more_spam.com (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id AAA25969 for "MicroscopyFilteredEmail1-at-msa.microscopy.com"; Sun, 2 Sep 2001 00:05:46 -0500 (CDT) Received: from gol-mas2.austar.net.au ([203.22.8.216]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id AAA25961 for {microscopy-at-sparc5.microscopy.com} ; Sun, 2 Sep 2001 00:05:34 -0500 (CDT) Received: from 150 (bds-228-109.tow.austar.net.au [203.21.228.109]) by gol-mas2.austar.net.au (Mirapoint) with SMTP id ADU82053; Sun, 2 Sep 2001 15:03:17 +1000 (EST) Received: by localhost with Microsoft MAPI; Sun, 2 Sep 2001 15:02:55 +1000 Message-ID: {01C133C0.5859C4A0.jim-at-proscitech.com} "jim-at-proscitech.com" {jim-at-proscitech.com} , "'Angela Klaus'" {avklaus-at-amnh.org} , "microscopy-at-sparc5.microscopy.com" {microscopy-at-sparc5.microscopy.com}
Reinhard Rachel also wrote on this back-channel and so I expect that you are right: I confused the early FE-SEM and FE-TEM. I saw the Siemens instrument in the Karlsruhe application lab in October 76. I was there for two other instruments but the large FE instrument was of course rather eye-catching. 25 years later I recall that it had a short, but large diameter column and so thought of it in the long-term as a FE-SEM. Hope that this was not a complete waste of time as these contributions have placed other FE instruments in a historical context. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Saturday, September 01, 2001 6:18 AM, Alan Nicholls [SMTP:nicholls-at-uic.edu] wrote: } Jim } } I think you mean FE-STEMs not FE-SEMs. The first VG HB5 STEM was delivered } in 1973 to University of London. } } Regards } } Alan } } At 09:55 AM 8/31/2001 +1000, Jim at Proscitech wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } In 1976 both, Siemens and Vacuum Generator marketed FESEMs. The Vacuum } } Generator unit was considerably cheaper and possibly better too. } } Incidentally, the Siemens unit became a fiasco and likely was a major reason } } for the company's board to abandon the building of electron microscopes a } } year } } or two later. Siemens had been the first commercial manufacturer of electron } } microscopes and built the best instruments in the 50th and early 60th. The } } Philips EM300 was the first serious challenge to Siemens. } } Michael Beer of Chicago published a great deal of FESEM developments in the } } early 70th. } } Cheers } } Jim Darley } } ProSciTech Microscopy PLUS } } PO Box 111, Thuringowa QLD 4817 Australia } } Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com } } Great microscopy catalogue, 500 Links, MSDS, User Notes } } ABN: 99 724 136 560 www.proscitech.com } } } } On Thursday, August 30, 2001 9:43 PM, Angela Klaus [SMTP:avklaus-at-amnh.org] } } wrote: } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } Hi Listers... } } } } } } Does anyone know when the first cold field emission microscope became } } } commercially available? } } } } } } Thanks and best, } } } } } } Angela } } } } } } Angela V. Klaus } } } } } } Director, Interdepartmental Laboratories } } } American Museum of Natural History } } } Central Park West at 79th Street } } } New York, NY 10024 USA } } } } } } Email: avklaus-at-amnh.org } } } Tel: 212-769-5977 } } } } } Alan W Nicholls, PhD } Director of Research Service Facility (Electron Microscopy) } Research Resources Center - East (M/C 337) } Room 100 Science and Engineering South Building } The University of Illinois at Chicago } 845 West Taylor St } Chicago, IL 60607-7058 } } Tel: 312 996 1227 } Fax: 312 996 8091 } Office: Room 110 } } Web site www.rrc.uic.edu
Dear Gary, I will receive my Axiopklan only on next week - so I have still no experience and cannot help. With best wishes Jiri Kalvoda ----- Original Message ----- } From: Gary Gaugler {gary-at-gaugler.com} To: {jsharp-at-zeiss.com} Cc: MSA listserver {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, August 22, 2001 5:09 AM
Here, here! (or is it hear , hear?) Ken Converse Quality Images
Paula Sicurello wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Listers, } } Nestor's little slip and his apology should make us realize how valuable Nestor and his service of the listserver is to us. I just want to say Thank You Nestor for all the hard work you do to keep us on-line and spam free. You help all of us to stay connected in a nice environment where we can ask our questions and communicate our ideas. } } Thank You Nestor! } } } Paula :-) } } Paula Sicurello } George Washington Univ. Medical Center } Dept. of Pathology, Ross Hall rm 505 } Electron Microscope Lab } 2300 Eye St. } Washington, DC 20037 } 202-994-2930 phone } 202-994-2518 fax } } } }
-----Original Message----- } From: John A. Reffner [mailto:JAReffner-at-compuserve.com]
On Tuesday, October 2, 2001, Dr. Joseph I. Goldstein will receive the New York Microscopical Society's Ernst Abbe Memorial Award at a special symposium in Atlantic City, New Jersey. The Symposium is part of the Eastern Analytical Symposium & Exposition. The session starts at 9:00-am with the presentation of the Abbe Award. The program includes:
Joseph I. Goldstein Advances in Scanning Electron Microscopy and X-Ray Microanalysis
Charles E. Lyman Quantitative X-Ray Microanalysis in the Environmental SEM
David B. Williams Microbeam Analysis of Metallic Meteorites
Eric Lifshin Pushing the Limits of Spatial Resolution in X-Ray Microanalysis
Come and join the celebration Dean Goldstein's contributions to science of microscopy and educating so many microscopists.
The EAS meeting will be held in the Atlantic City Convention Center, October 1-4, 2001. For more information contact the EAS Homepage:( http://www.eas.org).
I have run into a file format problem between Semper for Windows(image processing software from Synoptics, UK running on Microsoft Windows 98) and EMS (image simulation software by Prof Stadelmann running on a Silicon Graphics workstation). The EMS suite of programs for image simulation has a routine se1 that writes the R type images (created by the im1 routine, in the present case) in Semper file format which is essentially a Fortran unformatted output. This file is to be read into Semper using the unformatted option of the read command. However I get a
Message: illegal structure for unformatted file ?129: File I/O error 6419 on unit 1 file 'd:\users\divakar\semper\images\rd1082.unf'
error on Semper for Windows and a similar message on Semper for DOS version 6.4. I would appreciate help / advise from members of this list in this regard. Is this because the binary code on Unix and DOS /Windows is different? Or is this a version problem? Does somebody have software / image file / disc file format details which can be used to inter-convert between these operating systems and / or hardware?
With Best Regards, ---- Divakar R Physical Metallurgy Section, Indira Gandhi Centre for Atomic Research Kalpakkam 603102, India ----
We have an Electroscan ESEM E3, on which we have had different problems with the stage. In one case it was necessary to change some cables to fix the problem, but occasionally the stage won't move in the X-Y directions, and the problem is solved simply by changing the magnification on the microscope!! We haven't yet been able to come up with an explanation for this strange behaviour, but I hope for you, that your problems may be solved in this simple way.
Kind regards, Jesper ________________________________ Jesper Vejloe Carstensen Electron Microscopy & Microanalysis Risoe National Laboratory, Materials Research Department P.O. Box 49, DK-4000 Roskilde, DENMARK Phone: 46 77 57 76, Fax: 46 77 57 58 E-mail: jesper.v.carstensen-at-risoe.dk Web: www.risoe.dk/afm
-----Original Message----- } From: Gordon Vrololjak [mailto:gvrdolja-at-nature.Berkeley.EDU] Sent: 31. august 2001 18:24 To: Microscopy-at-sparc5.microscopy.com
Hello, I've been going through setting up an Electroscan ESEM E3 in our laboratory and vacuum problems aside, we now have a stage that doesn't want to move. Anyone ever come across stage problems and have some suggestions that I may try for this instrument?
The stage will actually move if you give it a gentle push along a gear arm, but it often locks up in the Z direction, and sometimes X/Y. Auto calibrate allows the stage to move in X/Y, but it never calibrates the Z direction. Thanks.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/ \ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
Dear fellow list server members, We have a Si(Li) detector with Be/UT window attached to a SEM. Some time before I started my job here there was a vacuum fault where pump oil backstreamed into the SEM column -my understanding is that the Be window was in position at the time. We are now seeing low counts for light elements when used in thin window mode and quantitative data reflect this, even though the oil streamed in when the Be window was being used. The manufacturer has suggested the likely source is ice on the crystal (from figures below). This problem I am assured only started after the vacuum/oil problem which indicates oil is the likely problem. I include K/L line data collected from a copper sample at 10, 15, 20 kV with both the Be and UT windows in place. The counts represent the peak windows/ROI of the K and L lines. What do you make of these figures below? As I have no reference data or anything to compare with. I would appreciate any input or suggestions you have.
Beam Window Cu L Cu K K/L ratio 20 Be 47461 156947 3.3 20 utw 84033 110570 1.3 15 Be 18357 24336 1.3 15 utw 34930 18172 0.5 10 Be 15225 1070 0.07 10 utw 28005 880 0.03
Mant thanks.
Martin Roe Electron Microscopist Materials/Engineering Department Wolfson Building Nottingham University University Park Nottingham NG7 2RD tel +44 (0) 115 9513768 tel =44 (0) 115 9513871 fax +44 (0) 115 9513764 email: martin.roe-at-nottingham.ac.uk
Due to you didn't mention anything about live time, beam condition, i.e, beam current, beam alignment, working distance and ect.. so it is quite difficult to suggest anything about your results. However, it seems likely that at 15kv (Be) and 20k(utw) are OK. Normally, at 20kV, K/L ratio is close to 1, it depends on the condition of the detector.
Try Ni K/L to confirm that, K/L ratio would be about 1 at 20 kV.
Regards,
Paiboob Nuannin Dept of Physics Faculty of Science Prince of Songkla University Hatyai 90110 Thailand
On Mon, 3 Sep 2001, Martin Roe wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } Dear fellow list server members, } We have a Si(Li) detector with Be/UT window attached to a SEM. Some time before I started my job here there was a vacuum fault where pump oil backstreamed into the SEM column -my understanding is that the Be window was in position at the time. We are now seeing low counts for light elements when used in thin window mode and quantitative data reflect this, even though the oil streamed in when the Be window was being used. The manufacturer has suggested the likely source is ice on the crystal (from figures below). This problem I am assured only started after the vacuum/oil problem which indicates oil is the likely problem. I include K/L line data collected from a copper sample at 10, 15, 20 kV with both the Be and UT windows in place. The counts represent the peak windows/ROI of the K and L lines. } What do you make of these figures below? As I have no reference data or anything to compare with. I would appreciate any input or suggestions you have. } } Beam Window Cu L Cu K K/L ratio } 20 Be 47461 156947 3.3 } 20 utw 84033 110570 1.3 } 15 Be 18357 24336 1.3 } 15 utw 34930 18172 0.5 } 10 Be 15225 1070 0.07 } 10 utw 28005 880 0.03 } } Mant thanks. } } Martin Roe } Electron Microscopist } Materials/Engineering Department } Wolfson Building } Nottingham University } University Park } Nottingham } NG7 2RD } tel +44 (0) 115 9513768 } tel =44 (0) 115 9513871 } fax +44 (0) 115 9513764 } email: martin.roe-at-nottingham.ac.uk } }
We tried a stretch alignment of the column to verify the basic astigmatism.
In low mag, specially in the central and lower magnification range (between 50 and 250) we see a ?blade? cutting the field of view only without any condenser aperture inserted.
This blade is the objective aperture support.
I suppose it is a normal thing, but I like to be sure about it.
Is the same on your machine?
If it is so, please let me know.
Thanks a lot!
P.S. Thanks Nestor!
Marco Arienti
__________________________________________________________________ Abbonati a Tiscali! Con VoceViva puoi anche ascoltare ed inviare email al telefono. Chiama VoceViva all' 892 800 http://voceviva.tiscali.it
We tried a stretch alignment of the column to verify the basic astigmatism.
In low mag, specially in the central and lower magnification range (between 50 and 250) we see a "blade" cutting the field of view only without any condenser aperture inserted.
This blade is the objective aperture support.
I suppose it is a normal thing, but I like to be sure about it.
Regarding my recent experiences with my used Axioplan, I consider the issue to be closed. Zeiss sent a factory technician to my site and corrected the problem at no charge.
I have little experience with SEM and EDX detector performance, however, I have used a range of Be, thin window and windowless detectors on TEMs and there are a couple of comments I would like to make.
If the degradation in performance occurred directly after the oil backstreaming then I agree that oil is quite likely to be the cause but otherwise it is quite possibly ice on the crystal.
Has the detector liquid nitrogen always been kept well filled? If the internal 'cryo pump' starts to warm up then you could get icing on the crystal.
Why not condition the detector anyway? Certainly on my Oxford Instruments (ex Link) detectors it is easy to do and does not seem to degrade the performance at all. I regularly condition the windowless detector on my JEOL2010 TEM (about every 6 months) to get the light element performace back. However, I have not had to condition the SATW detector (on another TEM) in over two years so I guess the water comes from the microscope and not the detector.
Regards, Ron
On Mon, 03 Sep 2001 17:02:31 +0100 Martin Roe {Martin.Roe-at-nottingham.ac.uk} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } Dear fellow list server members, } We have a Si(Li) detector with Be/UT window attached to a SEM. Some time before I started my job here there was a vacuum fault where pump oil backstreamed into the SEM column -my understanding is that the Be window was in position at the time. We are now seeing low counts for light elements when used in thin window mode and quantitative data reflect this, even though the oil streamed in when the Be window was being used. The manufacturer has suggested the likely source is ice on the crystal (from figures below). This problem I am assured only started after the vacuum/oil problem which indicates oil is the likely problem. I include K/L line data collected from a copper sample at 10, 15, 20 kV with both the Be and UT windows in place. The counts represent the peak windows/ROI of the K and L lines. } What do you make of these figures below? As I have no reference data or anything to compare with. I would appreciate any input or suggestions you have. } } Beam Window Cu L Cu K K/L ratio } 20 Be 47461 156947 3.3 } 20 utw 84033 110570 1.3 } 15 Be 18357 24336 1.3 } 15 utw 34930 18172 0.5 } 10 Be 15225 1070 0.07 } 10 utw 28005 880 0.03 } } Mant thanks. } } Martin Roe } Electron Microscopist } Materials/Engineering Department } Wolfson Building } Nottingham University } University Park } Nottingham } NG7 2RD } tel +44 (0) 115 9513768 } tel =44 (0) 115 9513871 } fax +44 (0) 115 9513764 } email: martin.roe-at-nottingham.ac.uk } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
I'll second what Jesper says. Our E3 also occasionally does the "lock-up" thing but can always be freed up simply by changing the magnification. Why this should fix it is, I think, one of the great mysteries of the universe. We've also occasionally experienced slow movement of the stage in the XY direction. This can make acquiring a digital image a real problem at times. Usually you can stop this drift by changing the pressure in the chamber.....Great Mystery of the Universe #2.....
Frank Thomas Geological Survey of Canada (Atlantic) Bedford Institute of Oceanography Dartmouth, Nova Scotia ----- Original Message ----- } From: "Gordon Vrololjak" {gvrdolja-at-nature.Berkeley.EDU} To: {Microscopy-at-sparc5.microscopy.com} Sent: Friday, August 31, 2001 1:23 PM
Charles Garber in his recent posting wrote: } Not all YAG's and } YAP's are created equal. So when making comparisons, and statements about } their scintillators, it would be helpful for one to state the brand of their } scintillator crystal. These crystals are certainly not a generic entity } where all are the same irrespective of the source.
I am a bit puzzled by this. I was under the impression that all of the YAG and YAP scintillators offered for sale in this country originate from the same ultimate supplier in Europe. Has this changed? Or perhaps there are different grades of crystal?
Fred Schamber Aspex LLC
"Garber, Charles A." wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Earl Weltmer and Brad Johnson wrote: } ============================================================ } } Hello, } } I need to replace the SE scintillator on my SEM (JEOL 5900), and I was } } considering using a YAP scintillator instead of the typical phosphorous } } scintillator. Does anyone have any experience or suggestions on this? } ++++Brad Johnson } } About ten years ago I ran some tests for signal on several scintillator } types: YAP, YAG, standard aluminum coated scintillator & "uncoated" } scintillator. } The result at that time were surprising. } } Using a JEOL 35C SEM & running the same specimen current & PMT voltage with } each scintillator the best signal was given by the original aluminum coated } scintillator. } } The second best was the "uncoated" scintillator & it was only 80% of the } signal the standard aluminum coated scintillator. } The YAP & YAG gave only 60% of the standard aluminum scintillator. } } I was very disappointed as the YAP & YAG were significantly more than the } standard scintillator. } The conclusion that I came to was that the YAP & YAG would probably give } more longevity & could justify their extra cost but they were not fit for } high resolution. The YAP & YAG would probably be suitable for probe work. } } Keep in mind this experiment was done about ten years ago. Perhaps the } crystals have improved in recent years & the data may need a re-evaluation. } I am sure that there is someone who would be willing to criticize my results } (maybe in Northern Calif.?) but this the data that I received at the time. } +++++ Earl Weltmer } ============================================================ } We have offered P-47, YAG and YAP scintillators for SEM applications for } more then twenty years. Earl is correct that the P-47 performance, **but } when first installed**, does probably give a superior performance. However } , not all YAG and YAP crystals come out of the "same cookie cutter." From } what we have been able to deduce, a high quality YAG or YAP single crystal } scintillator will perform so close to that of the P-47 that one usually has } difficulty detecting the difference. However, as everyone knows, the P-47 } once installed, in terms of performance, goes only down. The only question } is how fast it will go down. And that rate of deterioration of performance } depends among other things on the type of work being done. For example, the } higher x-ray fluxes associated with EDS (and especially WDS) tend to cause a } more rapid deterioration. } } Also, if Earl's experiment was done as described, and if a YAG was tested } against P-47 but using the same PMT, then one would expect to find an } inferior performance, because the PMT that is optimum for P47 is not the } same one that would have been optimum for YAG. Now this might be less of a } problem today than it would have been ten years ago, but if the PMT was not } changed, then the right test (IMHO) was not the one being performed. To } make the comparison, one would have had to have taken out the S11 style and } replaced it with an S20 PMT. This point is further explained on our website } URL } http://www.2spi.com/catalog/scintill/spi-yag.html } } When a YAG or YAP scintillator is installed, its performance level is } constant and does not deteriorate. So while the comparison might suggest } some superiority at the very beginning, in most instances that we have seen, } or have been led to believe, it is not long that the SEM running on YAG or } YAP in general, is operating at a higher level of performance. And the SEM } is not subjected to the downtime associated with the need to change a P-47 } powder scintillator from time to time. } } For those operating at low KV in the BSE mode (and using YAG or YAP), and } high magnifications, the image is clearly less noisy. A collection of ten } different references from different laboratories around the world, which } have been collected on URL } http://www.2spi.com/catalog/scintill/sem-tem.html } at least to some, pretty much document the superiority of the single crystal } scintillators over the powder scintillators. I guess one could always try } to do more studies, but anyone that is familiar with these publications } finds their conclusions pretty persuasive. } } We have generally recommended that the SPI single crystal scintillators } offered many advantages over the more traditional P-47 scintillator. We } have been under the impression that those who have made the switch have been } pretty happy with their results, though we do hear from time to time of } someone who would not agree with that statement. Of course, the performance } is related to the nature of the doping of the crystal, and as I said, not } all these crystals come out of the same cookie cutter. Not all YAG's and } YAP's are created equal. So when making comparisons, and statements about } their scintillators, it would be helpful for one to state the brand of their } scintillator crystal. These crystals are certainly not a generic entity } where all are the same irrespective of the source. } } Disclaimer: SPI Supplies is a major supplier of scintillators for SEM } applications including P-47's, YAGs and YAPs. } } Chuck } } =================================================== } Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 } President 1-(800)-2424-SPI } SPI SUPPLIES FAX: 1-(610)-436-5755 } PO BOX 656 e-mail: cgarber-at-2spi.com } West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com } } Look for us! } ############################ } WWW: http://www.2spi.com } ############################ } ==================================================
The first VG HB5 FE-STEM was delivered in 1973 to University of London.
Both AEI and Seimens got into dedicated STEM shortly before they pulled out of the EM market in the early 70's. VG was able to attract a number of the main characters who had worked at AEI to form the core of the VG Microscopes unit.
The first electron microscope VG produced was a W thermal sourced SEM (Miniscan) for which they were awarded the Queens award for technological innovation in 1970. They then went on to produce the HB5 High Vacuum FE-STEM and HB50 High Vacuum FE-SEM. All of the latter were sold with Auger spectrometers as a Scanning Auger Microprobe.
Regards
Alan
At 09:55 AM 8/31/2001 +1000, Jim at Proscitech wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Alan W Nicholls, PhD Director of Research Service Facility (Electron Microscopy) Research Resources Center - East (M/C 337) Room 100 Science and Engineering South Building The University of Illinois at Chicago 845 West Taylor St Chicago, IL 60607-7058
On both the E3 and 2020 the stage movement with the Joy Stick is linked to magnification. The higher the mag the slower the stage speed. Above about 8000 X the stage motors are disabled and beam shift is active with the Joy Stick.
The apparent movement of the image should remain the same at all mags. If the mag is above about 8KX then the amount of image movement is limited by the beam shift. To continue searching the magnification has to be lowered.
For ElectroScan ESEM questions (E2,E3,Explorer, 2010 or 2020) world wide contact me directly at dsimpson-at-ma.feico.com For XL ESEM (XLFEG ESEM, XLLab6 ESEM or XLTMP ESEM in North America contact me directly at dsimpson-at-ma.feico.com For XL ESEM (XLFEG ESEM, XLLab6 ESEM or XLTMP ESEM for the rest of the world contact their local Service group.
Regards Derek Simpson ESEM Technical Support Manager FEI Company One Corporation Way #2 Peabody, MA 01960 E-mail: dsimpson-at-feico.com Voice: 1.978.538.6700 Fax: 1.978.531.9648
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Dear Gordon,
We have an Electroscan ESEM E3, on which we have had different problems with the stage. In one case it was necessary to change some cables to fix the problem, but occasionally the stage won't move in the X-Y directions, and the problem is solved simply by changing the magnification on the microscope!! We haven't yet been able to come up with an explanation for this strange behaviour, but I hope for you, that your problems may be solved in this simple way.
Kind regards, Jesper ________________________________ Jesper Vejloe Carstensen Electron Microscopy & Microanalysis Risoe National Laboratory, Materials Research Department P.O. Box 49, DK-4000 Roskilde, DENMARK Phone: 46 77 57 76, Fax: 46 77 57 58 E-mail: jesper.v.carstensen-at-risoe.dk Web: www.risoe.dk/afm
-----Original Message----- } From: Gordon Vrololjak [mailto:gvrdolja-at-nature.Berkeley.EDU] Sent: 31. August 2001 18:24 To: Microscopy-at-sparc5.microscopy.com
Hello, I've been going through setting up an Electroscan ESEM E3 in our laboratory and vacuum problems aside, we now have a stage that doesn't want to move. Anyone ever come across stage problems and have some suggestions that I may try for this instrument?
The stage will actually move if you give it a gentle push along a gear arm, but it often locks up in the Z direction, and sometimes X/Y. Auto calibrate allows the stage to move in X/Y, but it never calibrates the Z direction. Thanks.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/ \ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
An alternative you may want to consider is the Polaron E6700 Evaporator, manufactured by Thermo VG Scientific. We are their distribution and service agent in the U.S. and would be happy to supply you with more information.
Best regards,
Michael R. Nesta General Manager Energy Beam Sciences, Inc. Agawam, MA USA Tel: (413) 786-9322 mnesta-at-ebsciences.com "Adding Brilliance to Your Vision"
-----Original Message----- } From: K.N. Bozhilov [mailto:bozhilov-at-citrus.ucr.edu] Sent: Friday, August 31, 2001 12:27 PM To: microscopy-at-sparc5.microscopy.com
Hi All,
I would greatly appreciate if you could share your opinion about performance and reliability of the Bench Top Turbo III vacuum evaporator from Denton Vacuum, also alternative suggestions are more than welcome. We are looking currently to purchase a vacuum evaporator for general EM preparation capable of carbon and metal evaporation, glow discharge and thickness control. Cleanliness of the vacuum is our main concern and especially the fact that the Bench Top model does not have LN trap.
Thank you,
Krassimir N. Bozhilov Central Facility for Advanced Microscopy and Microanalysis University of California Riverside, CA 92521
We are parting with a used oil diffusion pump (w/ LN2 Trap) that has approx. 2 years wear and tear. The pump has been in storage for the last 10 years. The pump was manufactured by DAIA vacuum Engineering corp, model # DPF4Zs. The mounting flange is approx. 7.25 inches with an 8 hole bolt pattern. It was removed from a Hitachi S-4000 FESEM for replacement by a cleaner turbo model. If you are interested please contact me off-line. The pump is free but you must assume all shipping costs. Thanks, jr
I will hazard one guess being unfamiliar with either of the programs or formats that you describe. I suspect that there is a difference in how the two systems format text files. Under DOS and Windows, text lines normally end with a carriage return and line feed characters, ASCII codes 13 and 10. Under UNIX, lines often end with one or the other, but not both; I think carriage return is the norm.
I know that WS-FTP has an option for converting text files to the proper standard when transferring between the two different platforms. It might be able to help, but you will need to tell it that the file are text. It ordinarily runs in auto-detect mode and determines file type based on the extension. Unknown extensions default to binary format. Presuming you have to ship your files anyway, this might eliminate the need for a translator program.
There are also a number of utilities around that could give you a look at the binary codes for your file to see what characters are being used to indicate the end of the line. I have some I could recommend on the PC side if you don't already have one. They are relatively available on the shareware sites and could help verify what I have said. Some even serve as editors so you could make changes to one file manually to see if this is the source of your problem.
Warren
At 04:16 PM 9/3/2001 +0530, you wrote:
} I have run into a file format problem between Semper for Windows(image } processing software from Synoptics, UK running on Microsoft Windows 98) } and EMS (image simulation software by Prof Stadelmann running on a } Silicon Graphics workstation). The EMS suite of programs for image } simulation has a routine se1 that writes the R type images (created by } the im1 routine, in the present case) in Semper file format which is } essentially a Fortran unformatted output. This file is to be read into } Semper using the unformatted option of the read command. However I get } a } } Message: illegal structure for unformatted file } ?129: File I/O error 6419 on unit 1 file } 'd:\users\divakar\semper\images\rd1082.unf' } } error on Semper for Windows and a similar message on Semper for DOS } version 6.4. I would appreciate help / advise from members of this list } in this regard. Is this because the binary code on Unix and DOS } /Windows is different? Or is this a version problem? Does somebody have } software / image file / disc file format details which can be used to } inter-convert between these operating systems and / or hardware? } } With Best Regards, } ---- } Divakar R } Physical Metallurgy Section, Indira Gandhi Centre for Atomic Research } Kalpakkam 603102, India } ----
---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
It is Ladd's belief that the ideal vacuum evaporator should include a mechanical and a diffusion pump to get you to the 10(-7) range and should have a LN trap. The evaporation system should have a protective shield for cleanliness. If you are doing chromium evaporation then a turbo pump would be required.
John Arnott
Disclaimer: Ladd Research manufactures and sells vacuum evaporation systems. --
**** Please Note Our New Address, Fax and Phone Numbers ****
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K.N. Bozhilov wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------.} } Hi All, } } I would greatly appreciate if you could share your opinion about performance } and reliability of the Bench Top Turbo III vacuum evaporator from Denton } Vacuum, also alternative suggestions are more than welcome. We are looking } currently to purchase a vacuum evaporator for general EM preparation capable } of carbon and metal evaporation, glow discharge and thickness control. } Cleanliness of the vacuum is our main concern and especially the fact that } the Bench Top model does not have LN trap. } } Thank you, } } Krassimir N. Bozhilov } Central Facility for Advanced Microscopy and Microanalysis } University of California } Riverside, CA 92521 } } Tel 909 787 2998 } Fax 909 787 4324
It is possible to break the 8mm glass, but it can be very frustrating. If the users want wider knives, try to talk them into purchasing a diamond knife for thick sections. It is less expensive than diamonds for thin sectioning, and I think it is a good buy because of time saved making and changing knives and attaching water catchers. The only downside is that once there is a nick in the edge it is there until resharpening time.
I would like to prepare 0.05 M Cacodylate buffer pH 7.4, but do not have the recipe and protocol necessary for its preparation. Would it be possible to help me out?
Thanks in advance.
Patrícia Reis ____________________________ Escola Superior Biotecnologia Universidade Católica Portuguesa Rua Dr. António Bernardino Almeida 4200-072 Porto PORTUGAL Tel.: 225580044 Fax.: 225090351 e.mail: {mailto:pmreis-at-mail.esb.ucp.pt} pmreis-at-mail.esb.ucp.pt
I would like to attend a course on microscopy and imaging techniques which would include the latest developments in this field. Apart from the ones at MBL, Cold Spring Harbor and Jim Pawley's course in June, does anyone know of others. Being quite keen, I would prefer something this winter but any suggestions would be highly appreciated.
Thank you Judy
Judy Trogadis Bio-Imaging Coordinator St. Michael's Hospital, 8Q 30 Bond St. Toronto, ON M5B 1W8
Dear All, A colleague recently inherited an IEC Minot Rotary Microtome. Does anyone have a manual or parts? It is missing the speciman holder, mounting bar for knife height adjustment and means of measuring knife angle.
Thanks in advance, Glen -- Glen MacDonald Microscopy and Imaging Facility University of Washington Core for Communication Research Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 glenmac-at-u.washington.edu (206) 616-4156 (206) 616-1828 fax ************************************************************************** C:} The box said "Requires Windows95 or better". So I bought a Macintosh. **************************************************************************
Could also be a processor related problem. There is a difference in the way the Intel and Motorola processors store information. One of the two writes the LSB (least significant byte) first, the other the MSB (most significant byte). The best bet would probably be to use a format that can be read by both. TIF, for example, can be normally read by both as the file itself contains information about the byte order.
Can you transform your images into TIF?
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Warren E Straszheim [mailto:wesaia-at-iastate.edu] Sent: Tuesday, September 04, 2001 10:31 AM To: divakar-at-igcar.ernet.in Cc: Microscopy-at-sparc5.microscopy.com
I will hazard one guess being unfamiliar with either of the programs or formats that you describe. I suspect that there is a difference in how the two systems format text files. Under DOS and Windows, text lines normally end with a carriage return and line feed characters, ASCII codes 13 and 10. Under UNIX, lines often end with one or the other, but not both; I think carriage return is the norm.
I know that WS-FTP has an option for converting text files to the proper standard when transferring between the two different platforms. It might be able to help, but you will need to tell it that the file are text. It ordinarily runs in auto-detect mode and determines file type based on the extension. Unknown extensions default to binary format. Presuming you have to ship your files anyway, this might eliminate the need for a translator program.
There are also a number of utilities around that could give you a look at the binary codes for your file to see what characters are being used to indicate the end of the line. I have some I could recommend on the PC side if you don't already have one. They are relatively available on the shareware sites and could help verify what I have said. Some even serve as editors so you could make changes to one file manually to see if this is the source of your problem.
Warren
At 04:16 PM 9/3/2001 +0530, you wrote:
} I have run into a file format problem between Semper for Windows(image } processing software from Synoptics, UK running on Microsoft Windows 98) } and EMS (image simulation software by Prof Stadelmann running on a } Silicon Graphics workstation). The EMS suite of programs for image } simulation has a routine se1 that writes the R type images (created by } the im1 routine, in the present case) in Semper file format which is } essentially a Fortran unformatted output. This file is to be read into } Semper using the unformatted option of the read command. However I get } a } } Message: illegal structure for unformatted file } ?129: File I/O error 6419 on unit 1 file } 'd:\users\divakar\semper\images\rd1082.unf' } } error on Semper for Windows and a similar message on Semper for DOS } version 6.4. I would appreciate help / advise from members of this list } in this regard. Is this because the binary code on Unix and DOS } /Windows is different? Or is this a version problem? Does somebody have } software / image file / disc file format details which can be used to } inter-convert between these operating systems and / or hardware? } } With Best Regards, } ---- } Divakar R } Physical Metallurgy Section, Indira Gandhi Centre for Atomic Research } Kalpakkam 603102, India } ----
---------------------- Warren E. Straszheim Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
Dear lister? Our Narishige WR-90 micromanipulator does not work properly because the the water inside the system has leaked out slowly. Does anyone in the list know how to refill it? Any suggestion is highly appreciated. Thanks, Long ----------------------------------- Miao, Long Dept of Biological Science 334 Bio. Unit1 Florida State University Tallahassee, FL32306-4370 email: lmiao-at-bio.fsu.edu Voice: (850)644-9817 FAX : (850)644-0481
Dear Listers, If any of you have removed a jammed film cassette from a JEOL 1200EXII, I would like to know how you dislodged it. We will eventually have a service technician to help but I cannot figure out how to remove the plate. I did manage to remove the film and yes the plate was inserted right side up. Thanks in advance. Rosemary
A one day short course on "Digital Image Capture and Management in Light Microscopy" by Dr. Mary McCann & Dr. John McCann is being offered on Sunday September 30,2001 at the Eastern Analytical Symposium and Exhbition in Atlantic City, NJ.
Don O'Leary
-----Original Message----- } From: Judy Trogadis [mailto:TrogadisJ-at-smh.toronto.on.ca] Sent: Tuesday, September 04, 2001 9:13 AM To: Microscopy-at-sparc5.microscopy.com Cc: Judy Trogadis
Fellow Microscopists:
I would like to attend a course on microscopy and imaging techniques which would include the latest developments in this field. Apart from the ones at MBL, Cold Spring Harbor and Jim Pawley's course in June, does anyone know of others. Being quite keen, I would prefer something this winter but any suggestions would be highly appreciated.
Thank you Judy
Judy Trogadis Bio-Imaging Coordinator St. Michael's Hospital, 8Q 30 Bond St. Toronto, ON M5B 1W8
I have experience many jams. Generally occurring with old age. Anyway, JEOL camera mechanisms are wonderful mechanical beasts. Two pushing nails in the mechanism catch the plates. If you look at the plates, on one side there are two small rectangles, which are cut out. The nails use these to capture the plates and push them through the mechanism. These nails, together with the plates wear down and eventually sometimes they miss and do not mate. Generally jamming in the open slot where the plates come out of the box.
So, to dislodge a jam there are several solutions:
1) Waggle the mechanisms in the hope it clears - not a good choice. 2) Make a piece of stiff plastic sheet the width of a plate and the length around 15". Try to insert the sheet above the canisters but below the top mechanisms so as to lift the nails free of the canisters. You can see the nails when you look into the camera above the boxes on both sides. They look like small arms with a piece of copper alloy on the end. Pull camera out if nails are free. 3) Remove the whole camera mechanism. Simply remove the bolts from below the camera chamber (6 I think), and then pull the camera mechanism out in one piece. The jam can then be freed by rotating the mechanism a wee bit.
If you have frequent jams, consider replacing the pushing nails (first) then the plates. Also a good idea to replace the Teflon sheet bearings, but get a JEOL engineer to do this otherwise the position of the mechanism could be lost.
Good luck,
Keith. ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Dr. K. Moulding.
Materials Characterisation and Preparation Facility Hong Kong University of Science and Technology, Clear Water Bay, Kowloon, Hong Kong.
I have gotten ahold of a Zeiss camera lucida drawing tube. It has a mirror at an angle that allows one to draw underneath the inclined eye tube. Works for a Zeiss Standard 16. I am getting the hang of it a bit. However, I have at least one question:
What is the collar for (referred to as "ring clamp" below)? It is somewhat eccentric, but I cannot imagine any purpose for it. The device doesn't fit over it, but nearly does. It *is* useful for attaching it to a wider eyepiece tube.
This plastic collar fits snugly inside the eyepiece tube of an Olympus SZH; this is good. Also, it fits over the end of the standard 16 eye tube, with a barrier that doesn't allow it to slip down over the tube. i have some documentation for a similar drawing tube, but there is a slight difference in the picture (there is a knurled knob at the point where the mirror/prism fits over the eyepiece. Mine doesn't have that, it fits snug). Here is the relevant section from the documentation for the pictured device:
1. Focus microscope.
2. Remove eyepiece and slip ring clamp on to tube. Replace eyepiece and fix with ring clamp. [I referred to the "ring clamp" as a collar.]
3. Clamp drawing prism on the eyepiece. [I assume this refers to the knurled screw that isn't present on my device---mine doesn't need to be clamped on.]
} From reading the description, though, I don't know what it is for. Is it a spacer to move the eyepiece up higher? Which Zeiss is it really designed for?
Also, I would greatly appreciate any tips. The tips I found in MICSCAPE were helpful. Can I initiate some correspondence with someone who has some experience with one of these fantastic devices?
Alan Davis Marianas High School
-- adavis-at-saipan.com 1-670-235-6580 Alan E. Davis, PMB 30, Box 10006, Saipan, MP 96950-8906, CNMI
I have steadily endeavored to keep my mind free, so as to give up any hypothesis, however much beloved -- and I cannot resist forming one on every subject -- as soon as facts are shown to be opposed to it. -- Charles Darwin (1809-1882)
Rosemary, I believe that the JEOL 1200 camera chamber is the same as the JEOL 100CX. You can slide out the film tray by making two metal strips measuring about 1" to 2" wide and about 10" to 12" long. Bend one end of each strip in the shape of an allen wrench to use as a sort of handle. These metal strips are used to raise up the two little arms that are locking up the film tray. While sitting on the floor so that the open camera chamber is at eye level and using a bright light you can look into the top area of the camera chamber just above where the film boxes are located. On each side and above the film boxes you will see these arms sticking down. You can use the metal strips to release these arms be sliding the metal strips straight in. Be sure that the film tray is all the way in so that there is no force on these arms holding them down. They are spring loaded and should raise up with only gently pushing on the metal strips. Be careful and do not apply a lot of force. You should be able to see these arms raise when you push in the metal strip. When both arms are raised the drawer should pull out easily. Again, don't use force. Hope that this helps.
John Humenansky/Staff Scientist Physical Electronics, Inc. (PHI) 6509 Flying Cloud Drive Eden Prairie, MN 55344 952-828-6387
Systran Federal Corporation in Dayton, Ohio, is managing an "on-site" Contract for the Materials and Manufacturing Directorate (ML), Air Force Research Laboratory (AFRL), at Wright Patterson Air Force Base, in Dayton, Ohio. Candidates are sought for a research position at AFRL/ML, and this position entails research and development on understanding surface forces and surface chemistry of MEMS devices and MEMS materials:
a) study surface forces of MEMS using scanning probe microscopes and special adhesion/friction testers as a function of environment (humidity, temperature, vacuum); develop an understanding of how surface forces, and hence friction and wear may be controlled using surface treatments including monolayers and solid coatings b) study surface chemistry, tribochemistry, and coating microstructure to elucidate the mechanisms controlling friction and wear in MEMS systems under extreme environments c) research lubricant transport behavior and dynamics in MEMS systems.
The successful candidate will have knowledge/experience of MEMS devices, surface chemistry, surface forces and tribology. Expertise in surface chemical probes such as scanning Auger microscopy, X-ray photoelectron spectroscopy, FTIR microscopy / grazing angle FTIR, micro Raman, and SEM/EDS are highly desirable. The candidate will use apparatus to study surface forces at the nano scale (scanning probe microscopes, special adhesion testers, and nano/micro friction measurement devices). Working knowledge of monolayer and solid coatings is also highly desirable. The focus of the research effort is to understand the chemistry and physics controlling friction and wear and to use that understanding to develop effective technologies leading to reliable MEMS operation in extreme environments. Analytical instrumentation is currently available, but the successful candidate must be capable of modifying instrumentation as part of the research program.
The advertised position is a full-time job with complete benefits (paid vacation, health and dental coverage, 401K, etc.). Interested candidates should respond to this advertisement by sending their resumes along with a listing of technical publications to the contact person listed below.
Dr. V. ("Nagu") Nagarajan V.P. & Program Manager Systran Federal Corporation 4027 Colonel Glenn Highway, Suite 210 Dayton, OH 45431-1672
We have an SEM application which requires an array of samples to be loaded into the microscope. For producing the array, we need to affix sticky tabs to an aluminum block (48 tabs in a 6 x 8 arrangement on a block about 3 inches by 4 inches).
Presently, we are producing these arrays by hand, but it would be much more efficient to produce the arrays in this form by simply pressing a sheet onto the block, then peel it off to give us all of the 48 tabs properly located on the plate in one shot.
Does anyone have a name of the manufacturer(s) of carbon sticky tabs, so we can contact them about this. Our supplier is not willing to provide this information.
On our search for high N.A. water-dipping/immersion objectives for 160 mm-corrected microscopes we came across some lenses made by a Russian company called 'LOMO Optics' (www.lomooptics.com), which we tried out on our Nikon Optiphot 2 two-photon scope, and on a Nikon Measurescope MM-11 (bottom illumination). Their performance is quite astonishing. And since we had to look for quite a while to find such lenses, we wanted to let the rest of the community know that you do not have to give away your old 160 mm-corrected scope yet. Prices are quite reasonable too. At LOMO they are willing to let you try out the lenses for yourself. But do not be surprised to get a phone call every day during that time. (And do not forget to negotiate!)
[1] Type EAF-A30-1, TT079, 30x water-immersion, N.A. 0.9, w.d. 1.3 mm, 0.17 mm coverslip correction (performance does not seem to suffer without coverslip), built-in iris, outer diameter: 0.545" at the tip for about 0.2", and 0.825" thereafter (the objective has a cap of that diameter, which (at least in our case) could be removed, giving an even smaller outer diameter of 0.710"), overall length when installed: ~1.050".
[2] Type 96001, 70x water-dipping (apparently no coverslip correction)., N.A. 1.23, w.d. 220 µm, outer diameter -at- the tip: 0.426" to 0.585" conical for about 0.2", 0.800" above,overall length when installed: 1.26".
[3] Type 83127, 85x water-immersion., N.A. 1.0, w.d. between 200 and 230 µm (depending on the coverslip correction setting, which ranges from 100 µm to 200 µm), outer diameter -at- the tip: 0.733" for about 0.2", 0.825" above, overall length when installed: 1.26".
[4] Apparently they also sell a 100x water-immersion lens by now, which we have not seen yet.
With best regards,
Axel
__________________________________
Axel Blau California Institute of Technology Division of Biology 156-29 1200 E. California Blvd. Pasadena, CA 91125
I was at a talk where the carbon sticky dots were first presented. I wish I could remember who was speaking. I know that SPI has the exclusive distribution rights to them. I believe that they were developed by a group at Dupont. I would check the literature and see if they published. Try back issues of the meeting abstracts. I think that the talk that I went to was about 10 years ago.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Sinkler, Wharton [mailto:WSinkler-at-uop.com] Sent: Wednesday, September 05, 2001 10:17 AM To: 'Microscopy-at-sparc5.microscopy.com' Cc: Kuechl, Dorothy
Dear Listers,
We have an SEM application which requires an array of samples to be loaded into the microscope. For producing the array, we need to affix sticky tabs to an aluminum block (48 tabs in a 6 x 8 arrangement on a block about 3 inches by 4 inches).
Presently, we are producing these arrays by hand, but it would be much more efficient to produce the arrays in this form by simply pressing a sheet onto the block, then peel it off to give us all of the 48 tabs properly located on the plate in one shot.
Does anyone have a name of the manufacturer(s) of carbon sticky tabs, so we can contact them about this. Our supplier is not willing to provide this information.
I can't help you get it out, but just wanted to warn you that it might be possible for you to get your fingers crushed or severed if you're fishing around in the chamber and the jam suddenly frees itself. BE CAREFUL. I'm not sure if this is the case with your model, but it was on an older JEOL that we had. I'd wait for the tech or at least get his advice before reaching in.
We had some that would get caught, and the only thing to do with them after they were freed was to toss them. Don't try to straighten them.
Sara
On Tue, 4 Sep 2001, Rosemary Walsh wrote:
} Date: Tue, 4 Sep 2001 20:23:07 -0500 } From: Rosemary Walsh {rw9-at-psu.edu} } To: Microscopy-at-sparc5.microscopy.com } Subject: removing jammed TEM film cassettes } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Listers, } If any of you have removed a jammed film cassette from a JEOL } 1200EXII, I } would like to know how you dislodged it. We will eventually have a service } technician to help but I cannot figure out how to remove the plate. I did } manage to remove the film and yes the plate was inserted right side up. } Thanks in advance. } Rosemary } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
The jamming of the plate drive mechanisms in electron microscopes is nothing new. I remember back in the 1960s when I was using our two JEM-6A TEMs in my classes on electron Microscopy. With an average enrollment of 25 to 30 students per term, coming from all disciplins across campus, and having all degrees of mechanical aptitude, plate jams were not an uncommon event. The easiest way to clear up a really bad plate jam in the JEM-6As was to reach inside the plate chamber and wiggle the plate carrier around a bit until things worked loose. Unfortunately, however, the chamber opening was too small for my hands to get in deep enough to take care of a bad jam, and so on several occasions I had to enlist the help of my 5-year-old son, who eventually became quite expert at taking care of the problem. Times do change, but digital cameras have their problems too!
Good luck, and keep smiling, WCB -- Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan 3062 Dow Bldg.; 2300 Hayward St. Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
Does anyone know where I could purchase a battery powered vibrating engraving tool? We have the corded type but I need a portable one to use out on a factory floor. Vendor replies welcome
Paul D. Nolan Electron Optics
Alcan International Limited Kingston Research and Development Centre P.O.Box 8400, 945 Princess Street Kingston, Ontario K7L 5L9
Hi, all I would like an advice. Can anyone suggest a better type of dishes to use with glutheraldehyde and/or osmium tetroxide during fixation time. Which bottles are better to store these fixatives? Many thanks.
Patricia ____________________________ Patrícia João Reis (PhD student) Escola Superior Biotecnologia Universidade Católica Portuguesa Rua Dr. António Bernardino Almeida 4200-072 Porto PORTUGAL Tel.: 225580044 Fax.: 225090351 e.mail: {mailto:pmreis-at-mail.esb.ucp.pt} pmreis-at-mail.esb.ucp.pt
"Fixed 2-5 minutes in Champy's fluid, rinsed 2x in water" . . . followed by ETOH dehydration, CPD, mounting and SEM viewing.
O.k., we give up, searched numberous books, papers, catalogs, web sites, etc. . . . does anyone have a recipe for "Chmapy's fluid" a.k.a. "Champy's Solution"? Please?
Thank you much!
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
} We had some that would get caught, and the only thing to do with them } after they were freed was to toss them. Don't try to straighten them. }
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-2131 University of Connecticut Storrs, CT 06269-2131 Phone: 860-486-3588 Fax: 860-486-6369
Anyone out there knows how to run up an X-ray film (exposed as well as unexposed) for TEM????? Neelima Shah.............. Biomedical Imaging Core Facility Uni of Pennsylvania Philadelphia, Pa.
LOMO (Leningrad's Opto-Mechanics Consortium) is well known manufacturer of the fine optics in the Russia. Their microscopes are cheap (relatively) and very suitable for students etc. LOMO also known as an manufacturer of the biggest single-piece 6 meters diameter glass mirror for the telescope situated in Caucasus mountains. That mirror was a biggest one in the world at the time of inventing. I believe, they did also first full-quartz-optic microscope for UV investigations and their MB-1/2 fluorescence microscopes with "opak-illumination" were great.
I don't have any financial interest in this company.
Sergey Ryazantsev
At 10:03 AM 9/5/01 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Ah, the wonders of a Google search! I found a wonderful histo manual at
http://131.229.114.77/histo/Manual.pdf
The recipe is given on p. 17 of this pdf but it is in code - the author lists standard histology stock solutions and then gives the recipes for a variety of fixatives by coding them from the stock solutions. Champy's is: "35G; 20K; 12M; 24Q(9)" which means: 35 ml of 1% chromic acid (aq.), 20 ml of 2% OsO4 (aq.), 12 ml of sat. potassium dichromate (9 g/ 100ml, aq.), 24 ml of distilled water, make up just before use. Sounds like nasty stuff.
I'm glad you asked - I'd have never found this pdf otherwise! (It is 113 pages and I haven't been able to figure out from where it originates - some histology course). If anyone knows who wrote it, I'd like to know!
Tamara
On Wed, 5 Sep 2001, Richard Edelmann wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } "Fixed 2-5 minutes in Champy's fluid, rinsed 2x in water" . . . followed by } ETOH dehydration, CPD, mounting and SEM viewing. } } } O.k., we give up, searched numberous books, papers, catalogs, web } sites, etc. . . . does anyone have a recipe for "Chmapy's fluid" a.k.a. } "Champy's Solution"? Please? } } Thank you much! } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 352 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } } "RAM disk is NOT an installation procedure." } }
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
I think I saw some at "Fredericks" out here on the West Coast. Contact me offline for the number.
Earl
----- Original Message ----- } From: {"Paul.Nolan-at-alcan.com"-at-sparc5.microscopy.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, September 05, 2001 12:42 PM
-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
Scott Walck wrote: =================================================== I was at a talk where the carbon sticky dots were first presented. I wish I could remember who was speaking. I know that SPI has the exclusive distribution rights to them. I believe that they were developed by a group at Dupont. I would check the literature and see if they published. Try back issues of the meeting abstracts. I think that the talk that I went to was about 10 years ago. =================================================== Let me shed some light on this: SPI Supplies was (and is) the exclusive licensee for the manufacturing and distribution of Tacky Dot™ Slides. The patent holder was DuPont and the inventor was Dr. Alan Cairncross, still at DuPont. I am guessing, but the paper you heard presented (at an MSA meeting ) which could have been about ten years ago (how time flies), probably was presented by Dr. Cairncross (and other authors all at DuPont). Full technical details are disclosed in US Patent # 5,356,751.
These should not be confused however with what are often times called by others as "carbon sticky dots" or "double sided conductive adhesive tabs". These are offered in various forms by SPI as well as the other main worldwide distributors of consumable supplies for SEM applications.
The original poster (Wharton Sinkler) asked about adhesive "tabs" but did not specify size. The largest dot size of the Tacky Dot Slides manufactured by SPI Supplies is 300 µm in an orthogonal array on 2000 µm centers. I think that Wharton was in essence asking about the smallest "carbon tab" or "disc" that is available but the smallest that I know of are are 9 mm and there are technical reasons why one could not go too much smaller than that (in our opinion).
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
from "Notes on microscopical technique for zoologists" CFA Pantin Cambridge Univ. Press 1962
Dr. Mel Dickson, Deputy Director, The Electron Microscope Unit, Adjunct Associate Professor, School of Microbiology & Immunology The University of New South Wales UNSW SYDNEY 2052 Australia. Phone +612 9385 6383 Fax +612 9385 6400 http://srv.emunit.unsw.edu.au/
The business of un-jamming JEOL plate camera doesn't have to be that gory. Just remove a fuse located a couple inches left of the camera door (make sure that little red light next to the fuse turns on), before reaching inside, and no fingers will be shed.
Assuming that the camera mechanism is not damaged- simplest thing to do to avoid jams- do not load film boxes to a full capacity(say, load 5 or 10 holders less), make sure that film holders are in perfect shape - ditto on Sara's comment, toss the damaged holders- and that the film holders are loaded correctly. Also, little Teflon rollers on the sides of the plate boxes must rotate freely (at a slight touch, no friction) and must have spotless surface (no defects, scratches, etc.). Replacement rollers are available from JEOL. If you are going to unscrew 6 screws underneath the camera compartment in order to pull the mechanism out (as was suggested in one of the previous messages)- beware that those screws are separating camera vacuum from the atmosphere. Make sure that the o-rings, screws, and seats are clean before replacing the screws, to avoid vacuum leak.
If the mechanism is damaged- repair options will depend on you mechanical aptitude. You are welcome to contact me off-line for further advice, yet I agree that your best bet is to contact JEOL service representative.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax
This message is made of 100% recycled electrons. -----Original Message----- } From: Marie E. Cantino {cantino-at-uconnvm.uconn.edu} To: Microscopy-at-sparc5.microscopy.com {Microscopy-at-sparc5.microscopy.com}
Reference: 'The Microtomist's Formulary and Guide' by Peter Gray. Blakiston Company, Inc., New York/Toronto. 1954:192 (794 pp.)
Cheers! Ken
-------------- Ken Tiekotter, Adjunct Professor The University of Portland Department of Biology 5000 N. Willamette Blvd. Portland, OR 97203
On Wed, 5 Sep 2001, Richard Edelmann wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } "Fixed 2-5 minutes in Champy's fluid, rinsed 2x in water" . . . followed by } ETOH dehydration, CPD, mounting and SEM viewing. } } } O.k., we give up, searched numberous books, papers, catalogs, web } sites, etc. . . . does anyone have a recipe for "Chmapy's fluid" a.k.a. } "Champy's Solution"? Please? } } Thank you much! } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 352 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } } "RAM disk is NOT an installation procedure." }
Does anyone know what has happened to the journal Scanning Microscopy?
The latest part we have is Volume 10 pt.4 1998 and our supplier cannot trace the publisher etc. all mail being marked return to sender. Has the journal ceased publishing?
Chris Jeffree
========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Daniel Rutherford Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JH, Scotland, UK Tel. #44 (0) 131 650 5345 FAX. #44 (0) 131 650 6563 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
Actually, the size of the tabs is not the issue. Sorry about the confusion.
What I was asking about are indeed double-stick carbon tabs about 1 cm diameter. I'd like to evenly space 48 of them on an aluminum plate, and am looking for a less tedious way than by hand.
An alternative would be to simply cover the entire plate with a double-stick conductive carbon layer, then depsoit the samples in the proper locations. This may be cheaper than getting something custom made. Does anyone know who might offer unperforated double stick conductive carbon material in large dimensions (about 3 x 4 inches)?
Thanks, Wharton
} -----Original Message----- } From: Garber, Charles A. [SMTP:cgarber-at-2spi.com] } Sent: Wednesday, September 05, 2001 9:20 PM } To: MICROSCOPY BB } Subject: Sticky dots and Tacky Dots } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Scott Walck wrote: } =================================================== } I was at a talk where the carbon sticky dots were first presented. I wish } I } could remember who was speaking. I know that SPI has the exclusive } distribution rights to them. I believe that they were developed by a } group } at Dupont. I would check the literature and see if they published. Try } back issues of the meeting abstracts. I think that the talk that I went } to } was about 10 years ago. } =================================================== } Let me shed some light on this: SPI Supplies was (and is) the exclusive } licensee for the manufacturing and distribution of Tacky Dot(tm) Slides. } The } patent holder was DuPont and the inventor was Dr. Alan Cairncross, still } at } DuPont. I am guessing, but the paper you heard presented (at an MSA } meeting } ) which could have been about ten years ago (how time flies), probably was } presented by Dr. Cairncross (and other authors all at DuPont). Full } technical details are disclosed in US Patent # 5,356,751. } } These should not be confused however with what are often times called by } others as "carbon sticky dots" or "double sided conductive adhesive tabs". } } These are offered in various forms by SPI as well as the other main } worldwide distributors of consumable supplies for SEM applications. } } The original poster (Wharton Sinkler) asked about adhesive "tabs" but did } not specify size. The largest dot size of the Tacky Dot Slides } manufactured } by SPI Supplies is 300 µm in an orthogonal array on 2000 µm centers. I } think that Wharton was in essence asking about the smallest "carbon tab" } or } "disc" that is available but the smallest that I know of are are 9 mm and } there are technical reasons why one could not go too much smaller than } that } (in our opinion). } } Chuck } } } =================================================== } Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 } President 1-(800)-2424-SPI } SPI SUPPLIES FAX: 1-(610)-436-5755 } PO BOX 656 e-mail: cgarber-at-2spi.com } West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com } } } Look for us! } ############################ } WWW: http://www.2spi.com } ############################ } ================================================== }
Thank you for all who replied we now have a recipe to try out.
Champy's:
=========
1% chromic acid 7 ml 3% K2Cr2O7 7 ml 2% OsO4 4 ml
from "Notes on microscopical technique for zoologists" CFA Pantin Cambridge Univ. Press 1962
========
"[3% aqueous potassium dichromate; 1% aqueous chromic acid; 2% aqueous solution of osmic acid (osmium tetroxide); (2:2:1)]" Page 804-805 Samuel Thompson's "Selected Histochemical and Histopathological Methods", 1966.
==========
"35G; 20K; 12M; 24Q(9)" which means: 35 ml of 1% chromic acid (aq.), 20 ml of 2% OsO4 (aq.), 12 ml of sat. potassium dichromate (9 g/ 100ml, aq.), 24ml of distilled water, make up just before use. Sounds like nasty stuff.
===========
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
Does anyone out there know: a) where to buy this stuff? b) How to make this stuff? or c) where I can find any articles, mentions or instructions on the use of this stuff? (other than the Fisheries Science article that comes up in Google) I've been told that it's a useful marker for the aragonite polymorph of CaCO3. (Thanks, John Hunt at CCMR!) Any feedback, however oblique, would be appreciated.
Jeff
-- Jeff Hedlesky Aladdin Companies 151 N. Main, Suite 700 Wichita, KS 67202
It's been a while since I've been on the list, and since I've been working as a microscopist.
If I image a sample in the SEM with the sample tilted, then want to make measurements from the images, what is the equation to use to correct for the tilt? I know it involves the cosine of the tilt angle, but beyond that, I'm drawing a blank.
Thanks for your assistance.
Nancy Zjaba, Technical Staff Alfalight Inc. 1832 Wright Street Madison, WI 53704 (608) 240-4875 nzjaba-at-alfalight.com
Thanks to the two of you for the information. The website -at- www.comet.net/gek/ is very interesting, and I did not know that there was some other distributor in the US. I have to agree that the quality of the lenses we got is outstanding for the price we paid.
With best regards,
Axel
----- Original Message ----- } From: Jeff Hedlesky To: Sergey Ryazantsev Cc: Axel Blau ; Microscopy-at-sparc5.microscopy.com Sent: Thursday, September 06, 2001 6:56 AM
For a flat sample with tilted to an angle, theta, the image perpendicular to the tilt axis is foreshortened by cos(theta). Take your measurement on the tilted image and divide by cos(theta).
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Nancy Zjaba [mailto:nzjaba-at-alfalight.com] Sent: Thursday, September 06, 2001 12:18 PM To: 'microscopy-at-msa.microscopy.com'
Hello to the list.
It's been a while since I've been on the list, and since I've been working as a microscopist.
If I image a sample in the SEM with the sample tilted, then want to make measurements from the images, what is the equation to use to correct for the tilt? I know it involves the cosine of the tilt angle, but beyond that, I'm drawing a blank.
Thanks for your assistance.
Nancy Zjaba, Technical Staff Alfalight Inc. 1832 Wright Street Madison, WI 53704 (608) 240-4875 nzjaba-at-alfalight.com
After tilting, the feature you want to measure will form the hypotenuse of a triangle. The horizontal projection you see will be the side of the triangle adjacent to the tilt angle. One side of the feature will be elevated with respect to the other and that amount will be the length of the side opposite the tilt angle.
The cosine of an angle is the ratio of the length of the adjacent side to the length of the hypotenuse. Therefore, you would use the following formula.
hypotenuse = adjacent/cosine, or
real length - observed length/cosine(tilt angle).
At 11:18 AM 9/6/2001 -0500, you wrote:
} Hello to the list. } } It's been a while since I've been on the list, and since I've been working } as a microscopist. } } If I image a sample in the SEM with the sample tilted, then want to make } measurements from the images, what is the equation to use to correct for the } tilt? I know it involves the cosine of the tilt angle, but beyond that, I'm } drawing a blank. } } Thanks for your assistance. } } Nancy Zjaba, Technical Staff } Alfalight Inc. } 1832 Wright Street } Madison, WI 53704 } (608) 240-4875 } nzjaba-at-alfalight.com
I was working with JEOL microscopes (100B, 100C, 100CX, 1200EX) for a few decades. Over all this time I do remember just a few accidents related to the camera mechanics in the microscopes. In all that cases the "human factor" was involved and usually the problem was improperly loaded into cassette the piece of the film. If film is not in place, sometime it cover one of the square slots on the cassette and "fork" could not catch the cassette properly. In this case the forks usually stick in the gap between two set of rails in the camera mechanism. As it was mentioned in the most replies, the solution is to move up those forks out of the gap. Usually I am doing so using flexible metal ruler with fuse OFF (great comment Vitaly). The bad things about JEOL camera design that it has extremely powerful motor with huge gear which is able to bent and ruin all rails in the camera if working improperly. Magically, it was never happens with my microscopes, but when I look inside the camera and see that huge gears I am worrying anyway.
Sergey
At 12:12 AM 9/6/01 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Wharton Sinkler wrote: ================================================= Actually, the size of the tabs is not the issue. Sorry about the confusion.
What I was asking about are indeed double-stick carbon tabs about 1 cm diameter. I'd like to evenly space 48 of them on an aluminum plate, and am looking for a less tedious way than by hand.
An alternative would be to simply cover the entire plate with a double-stick conductive carbon layer, then depsoit the samples in the proper locations. This may be cheaper than getting something custom made. Does anyone know who might offer unperforated double stick conductive carbon material in large dimensions (about 3 x 4 inches)? ================================================== We at SPI Supplies offer as a standard off-the-shelf product a sheet that is 500mm x 500 mm. You could cut out from that standard size sheet whatever size you might require. This is described on URL http://www.2spi.com/catalog/spec_prep/cond_adhes-sheets.html
This is the "master material" from which the double sided conductive carbon adhesive discs are fabricated.
We have heard of another customer who has made a template in the orthogonal arrangement required and then by hand is affixing (on large sheets) what they want to attach through the use of the template, which is suspended slightly above the adhesive and never actually contacts the adhesive. That would be the cheapest of the various alternatives.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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I dissolve 1g OsO4 in 25ml double distilled water and store it in a bottle with a Teflon lined screw cap. The bottle is then held in the refrigerator in a small plastic jar (available from all lab distributors - mine from Fisher or S/P) with a screw cap . In this way, the plastic becomes osmicated NOT the refrigerator. I have done this for years with no untoward results.
I fix with aldehydes in screw cap scintillation vials (20ml or 5ml) glass or plastic. Osmication is done in glass vials with Teflon lined screw caps using a minimum of OsO4 to do the job.
I make glutaraldehyde from a 10ml vial in dd HOH (50%, 10ml Glu + 40ml dd HOH yields 10% Glu) as a 10% solution. This in a screw top bottle with a Teflon lined cap. I then make up an appropriate amount of fixative using the 10% glu and 10X buffer concentrate where possible. The 10% Glu is kept in a plastic jar in the refrigerator.
I find that double containers are safe and reduce contamination considerably.
Hope this helps.
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II South Church Street West Chester, PA, 19383 eMail: fmonson-at-wcupa.edu http://darwin.wcupa.edu/casi/
Hello everyone, I have a researcher here that is having some difficulty with getting good histology at the EM level. She is fixing Drosophlia brains with 4%para/0.5% glut in phosphate buffer pH7.2, no osmium post fix, ethanol dehaydration, propylene oxide as a transitional solvent to polybed 812, then performing post sectioning immunolabeling. The tissue has "holes" in it when viewed at the TEM. They are not holes in the section, but appear to be voids in the tissue. We have tried leaving out the propylene oxide, we have osmicated, and have looked at osmotic strength of the fixative (and many variations of the above, to no avail). Can anyone offer advice? Randy Nessler Univ. of Iowa
} -----Original Message----- } From: Nancy Zjaba [mailto:nzjaba-at-alfalight.com] } Sent: Thursday, September 06, 2001 11:18 AM } To: 'microscopy-at-msa.microscopy.com' } Subject: SEM: Tilt correction factor } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } Hello to the list. } } It's been a while since I've been on the list, and since I've } been working } as a microscopist. } } If I image a sample in the SEM with the sample tilted, then } want to make } measurements from the images, what is the equation to use to } correct for the } tilt? I know it involves the cosine of the tilt angle, but } beyond that, I'm } drawing a blank. } } Thanks for your assistance. } } Nancy Zjaba, Technical Staff } Alfalight Inc. } 1832 Wright Street } Madison, WI 53704 } (608) 240-4875 } nzjaba-at-alfalight.com } } } }
Recently, a collegue was having problems with a white crystaline substance covering his samples of ear tissue after they were removed from the critical point dryer. It was believed to be from a contaminant in the ethanol and it was recommended to switch to acetone dehydration as it was less likely to have this problem. I haven't done SEM in years, so I haven't been paying attention to anything like this on this listserve. I was wondering if other people experiencing this problem. If you are using acetone, have you ever seen this?
Depends upon the geometry of the sample: try making the calculations for a sphere at a 45 degree angle.
My guess is that the measurements would be the same as if the sphere were at 0 degrees or 90 degrees.
For purely planar sample the equations would be valid.
regards,
Earl
----- Original Message ----- } From: "Nancy Zjaba" {nzjaba-at-alfalight.com} To: "'microscopy-at-msa.microscopy.com'" {microscopy-at-sparc5.microscopy.com} Sent: Thursday, September 06, 2001 9:18 AM
Karen,
One possible source for this contaminant is from the use of drying agents such as molecular sieves or other crystals. Unless one is extremely careful, the fine crystals get suspended in the liquid phase and dry down onto the specimen. We no longer use drying agents in our ethanol but use absolute ethanol directly from small (400-500 ml) bottles. If the bottles have been opened for awhile, then we use the alcohol to prepare a dehydration series since it is no longer anhydrous.
You CAN still use molecular sieves, etc. but we prepare them one month before use and take care not to jostle the bottles to any extent.
Another source for the crystalline material may be from prior contamination of the chamber by someone who had a dirty sample or contaminated ethanol.
JB
} Recently, a collegue was having problems with a white crystaline } substance covering his samples of ear tissue after they were removed } from the critical point dryer. It was believed to be from a } contaminant in the ethanol and it was recommended to switch to acetone } dehydration as it was less likely to have this problem. I haven't done } SEM in years, so I haven't been paying attention to anything like this } on this listserve. I was wondering if other people experiencing this } problem. If you are using acetone, have you ever seen this? -- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ##############################################################
The address for your LOMO dealer is: Microscopy Labs Box 338 Red Bank, NJ 07701 732 747 6228 fax 732 758 9142 ----- Original Message ----- } From: "Bradley Starcevich" {microscopist-at-excite.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, September 06, 2001 10:17 AM
Once again, my plea for help to you friendly and knowlegeable folks on this listserve produced twelve hints, warnings to protect my fingers and encouragement to keep smiling through equipment hassles. I have compiled them into a lengthy word document and would be happy to send it to anyone requesting it. The following were the most important bits of advice I received from all of you and from a discussion with one of JEOL's excellent service techs working out of Pittsburgh a) the camera mechanics are pretty much the same (100, 1200, 2010 even) b) if an error message indicating the "no film loaded" appears on the computer screen, remove the fuse which turns the mechanism's motor off (preventing the motor from burning out and your fingers should you elect to remove film + plate) c) examine plates--this one was "bowed" and showed a wear pattern d) don't load the film box to capacity e) dispose of damaged plates, even those with minor bends f) examine the teflon rollers on the film boxes, replace if they do not rotate freely
Thanks again to: Sergey Ryazantsev, UCLA; Robin Cross, Rhodes University, South Africa; Keith Moulding, Hong Kong University of Science and Technology, Hong Kong; Deb Stenzel Queensland Univ. of Tech, Brisbane, Australia: . John Hunt, Cornell: John Humenansky/Staff Scientist Physical Electronics, Inc. (PHI); Malcolm Haswell, UK; Richard Beanland, Structural Analysis Lab,Caswell Technology; Sara Miller, Duke University; Wilbur C. Bigelow, University of Michigan; Marie E. Cantino, University of Connecticut; Vitaly Feingold Scientific Instruments and Applications.
While we are on the topic of making up fixatives I have a supplementary question.
How do you open glass osmium tetroxide ampoules?
I put mine in a 100ml glass bottle and then break it with a glass rod. This is nerve-wracking, tedious (they do not break readily) and not as safe a procedure as I would like.
The ampoules used to have a weakened neck which could be sawn off but they do not come in this form from our supplier any more.
Is there an easier way?
Dave
On Thu, 6 Sep 2001 15:46:24 -0400 "Monson, Frederick C." {fmonson-at-wcupa.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I dissolve 1g OsO4 in 25ml double distilled water and store it in a bottle } with a Teflon lined screw cap. The bottle is then held in the refrigerator } in a small plastic jar (available from all lab distributors - mine from } Fisher or S/P) with a screw cap . In this way, the plastic becomes } osmicated NOT the refrigerator. I have done this for years with no untoward } results. } } I fix with aldehydes in screw cap scintillation vials (20ml or 5ml) glass or } plastic. Osmication is done in glass vials with Teflon lined screw caps } using a minimum of OsO4 to do the job. } } I make glutaraldehyde from a 10ml vial in dd HOH (50%, 10ml Glu + 40ml dd } HOH yields 10% Glu) as a 10% solution. This in a screw top bottle with a } Teflon lined cap. I then make up an appropriate amount of fixative using } the 10% glu and 10X buffer concentrate where possible. The 10% Glu is kept } in a plastic jar in the refrigerator. } } I find that double containers are safe and reduce contamination } considerably. } } Hope this helps. } } Fred Monson } } Frederick C. Monson, PhD } Center for Advanced Scientific Imaging(CASI) } West Chester University of Pennsylvania } Schmucker Science Center II } South Church Street } West Chester, PA, 19383 } eMail: fmonson-at-wcupa.edu } http://darwin.wcupa.edu/casi/ } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Using clean ethanol (does not need to be "dried" for CPD) would be a lot easier than replacing seals - if the valves in your CPD don't like acetone. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Ph +61 7 4774 0370 Fax:+61 7 4789 2313 service-at-proscitech.com Great microscopy catalogue, 500 Links, MSDS, User Notes ABN: 99 724 136 560 www.proscitech.com
On Friday, September 07, 2001 7:47 AM, Karen Pawlowski [SMTP:kpawlow-at-swbell.net] wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, } } Recently, a collegue was having problems with a white crystaline } substance covering his samples of ear tissue after they were removed } from the critical point dryer. It was believed to be from a } contaminant in the ethanol and it was recommended to switch to acetone } dehydration as it was less likely to have this problem. I haven't done } SEM in years, so I haven't been paying attention to anything like this } on this listserve. I was wondering if other people experiencing this } problem. If you are using acetone, have you ever seen this? } } Thanks for your comments, } } Karen Pawlowski, Ph.D.
I would first like to thank the list members for the responses I have received. Thanks are also due to software support at Synoptics for a prompt and informative reply to my e-mail. I provide here a brief description of how the problem was resolved finally, in the hope that the info may be useful to somebody someday, just as I have learned a lot from this list.
On checking the binary file from the SGI computer in a hex viewer, I confirmed that the bytes were not in the format required by Semper for a Fortran unformatted file. Also confirmed that this was not due to file transfer by explicitly setting the transfer mode in ftp to binary. The e-mail from Synoptics informed that the Fortran unformatted format was not intended to be portable across operating systems and different hardware and that the input / output commands in Semper supersede the read / write commands. I discovered that the format is not even portable from the DOS version to the Windows version of Semper. The main suggested solution was to convert the image to one of the standard binary image formats such as tiff that can be read into Semper.
Two solutions are possible: (i) the tv1 program in EMS can be used to display the EMS image and the screen can be exported to Semper format. This export file has the extension eix and is an ASCII file with the image pixel data as integers. When read into Semper it is available as a byte class image. Problem with this approach is that the pixel data floating point values are converted to 8-bit values and I am not sure if this affects further quantification. Also, it requires selecting the area to be saved and I tend to end up with odd sized images which included bits of the screen background requiring further cropping with Semper.
(ii) To change the source code of se1 such that all unformatted Fortran write statements are replaced with formatted write with the format strings as per Semper file format specifications. On compiling and running, this results in an ASCII file which can be read into Semper as a floating point image, that is, without any change in the pixel values. This is the solution I have adopted. The resulting file size is larger by about 3.5 times but this is not a limitation for me. I am still left with the minor problem that the title of image is getting written as a string of blank characters. My Fortran is a bit rusty and I hope to solve this soon.
Best Wishes to all.
-----Original Message----- } From: Divakar [SMTP:divakar-at-igcar.ernet.in] Sent: Tuesday, September 04, 2001 10:33 AM To: 'Microscopy-at-sparc5.microscopy.com'
Hi Alan, Go get em! I have a Camera Lucida for a Leitz Ortholux that I have sitting next to me in my office. The collar that I have fits over the narrow diameter eyepiece tube on my scope and sets my Camera Lucida at the exit pupil of the ocular. This permits very close concordance of the two images transmitted by the prism. Of course, my camera Lucida is an old thing with a 4x6" mirror hanging off to the right. My drawing surface must be inclined, because my oculars are inclined on my binocular headed microscope. I had to construct the inclined table, but it works anyway. Here's a project. Take a piece of paper and mark it off in half centimeters (i.e., every 5mm, Ho! Ho!). Using a high mag objective look at something small like a diatom. Using the graduations on the fine focus knob change focus by one graduation and move the paper one 5mm increment drawing the outline of the in-focus diatom at each point. You will quickly see how to limit your lines so they don't overlap. Try to predict what the result will be after you draw several outlines. The key to getting everything right is to balance the illumination of the paper with that of the scope, but you probably noticed that already. Send me a photo by direct eMail. The listserver doesn't convey attachments.
You are welcome to ask questions any time or to discuss, What kind of scope(S) do you have? Illuminators? Use high intensity on the paper!
Regards,
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II CASI Home Page: http://darwin.wcupa.edu/casi/ South Church Street West Chester, PA, 19383 Office: SS024 Phone: 610-738-0437 FAX: 610-436-3036 eMail (Work): fmonson-at-wcupa.edu eMail (Home): monson_fc-at-msn.com Please call before visiting
} ---------- } From: Alan Davis } Sent: Wednesday, September 5, 2001 7:16 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Camera Lucida drawing tube (Zeiss) questions } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I have gotten ahold of a Zeiss camera lucida drawing tube. It has a } mirror at an angle that allows one to draw underneath the inclined eye } tube. Works for a Zeiss Standard 16. I am getting the hang of it a bit. } However, I have at least one question: } } What is the collar for (referred to as "ring clamp" below)? It is } somewhat eccentric, but I cannot imagine any purpose for it. The device } doesn't fit over it, but nearly does. It *is* useful for attaching it to } a wider eyepiece tube. } } This plastic collar fits snugly inside the eyepiece tube of an Olympus } SZH; this is good. Also, it fits over the end of the standard 16 eye } tube, with a barrier that doesn't allow it to slip down over the tube. i } have some documentation for a similar drawing tube, but there is a slight } difference in the picture (there is a knurled knob at the point where the } mirror/prism fits over the eyepiece. Mine doesn't have that, it fits } snug). Here is the relevant section from the documentation for the } pictured device: } } 1. Focus microscope. } } 2. Remove eyepiece and slip ring clamp on to tube. Replace eyepiece } and fix with ring clamp. [I referred to the "ring clamp" as a } collar.] } } 3. Clamp drawing prism on the eyepiece. [I assume this refers to the } knurled } screw that isn't present on my device---mine doesn't need to be } clamped on.] } } } } } From reading the description, though, I don't know what it is for. Is it } a spacer to move the eyepiece up higher? Which Zeiss is it really } designed for? } } Also, I would greatly appreciate any tips. The tips I found in MICSCAPE } were helpful. Can I initiate some correspondence with someone who has } some experience with one of these fantastic devices? } } Alan Davis } Marianas High School } } -- } adavis-at-saipan.com 1-670-235-6580 } Alan E. Davis, PMB 30, Box 10006, Saipan, MP 96950-8906, CNMI } } I have steadily endeavored to keep my mind free, so as to give up any } hypothesis, however much beloved -- and I cannot resist forming one } on every subject -- as soon as facts are shown to be opposed to it. } -- Charles Darwin (1809-1882) } } } } } }
According to Prof Bigelow, one clears such a jam with a wiggle! But from the Eastern Hemisphere and Dr. Moulding we hear that such jams are freed by a waggle. Now what needs to be established is whether one should wiggle or waggle, or whether one must learn to do both, AND whether TEM's behave differently toward wiggling and waggling in different places (We all know JEOL is 100V no matter where you find it.). Having been raised in North America, I have been exposed to what has always been described as typical wiggling, and I would hope that if I ever saw someone waggle, I would know that waggling was being done. One might be terribly embarrassed if one were to characterize a wiggle as a waggle. For example, is there wiggling or waggling in typical down-under dancing? If one stands in Hong Kong ogling the girls as they pass by, is one traumatized by the wiggle or the waggle? We know that one can wiggle a nose, but conversely one can only wag one's tail - in North America. But then, we all have seen buns wiggle. Perhaps the relationship is something like this. A waggle could be a small wag, and since we know that Prof. Bigelow was not suggesting that one should wag the plate to free it, rather he suggested that we just give it a little wiggle. In this way one could assert no difference between a wiggle and a waggle, both being, similarly, small wags. One would not say, "Wag the plate a little." But one could "Waggle it!" or just "wag it a little!" This would all be so much more clear if dogs in North America were known to "wig their tails" when they are happy. Perhaps what we have is an hemispheric etymological inversion such that in the West, wiggle is to wag, where as, in the East, waggle is to wig! We here in the Western Hemisphere have learned to watch worms wiggle, but what do they do elsewhere, I wonder? Might a waggle merely be out of phase with a wiggle? Sine! Cosine! Do these differences occur, because of the differences between 50 and 60 Hz? Perhaps there might be NSF or WHO money for such a question.
Oh well, ruminating like a mad sow, he demurred, "No rumen in the old sow at all."
If anyone can clear any of this up, I certainly would like to know!
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II South Church Street West Chester, PA, 19383 eMail: fmonson-at-wcupa.edu http://darwin.wcupa.edu/casi/
While I have usually come to a comfortable resolution with respect to this kind of safety issue, this one has proved elusive. So.....
My normal procedure is to unwrap and clean the ampoule as I would my best glassware. That is, I treat it to a "biochemistry lab" cleaning, finishing with 3X rinses in sgdd(single glass-distilled deionized) HOH. Between the last two waters AND WEARING FRESH POWDER-FREE GLOVES, I score the ampoule three times with an ampoule file while holding it on a folded paper towel in the bottom of a shallow heavy-glass bowl. I then transfer the ampoule to an appropriate 100ml bottle (I use a serum bottle, because it is constructed of thick glass, shows imperfections readily (especially around the bottom), and can withstand the breaking of the ampoule). The scored ampoule breaks more readily than an unscored one, and three scores (~1cm apart) provide sufficient weakness for breakage to occur with relative ease. To further facilitate breakage, I add only some of the total amount of solvent HOH (for me, 25ml) sufficient to keep the ampoule from floating while providing enough for initial immersion of the crystals. Finally, I rest the serum bottle on the same folded paper towel in the same thick glass bowl to dissipate the shock of the breaking regimen. NOTE: Since it is at this part of the process at which failure has occurred (twice in 25 years the bottom of the bottle has separated from the barrel), I am still working in the hood, but with a substantial amount of 95% EtOH at hand to render the OsO4 harmless if I have an accident.
If the above smacks of science, please forgive the allusion. The method and approach are the result of what has evolved over time through trial and error and the application of common sense to a continuing, natty problem.
I have always hoped that someone would supply a FINAL solution so we can all be more comfortable when "Stock solution" time arrives.
Regards,
Fred Monson
} ---------- } From: Patton, David } Sent: Friday, September 7, 2001 5:30 AM } To: Monson, Frederick C. } Cc: 'Microscopy Listserver' } Subject: Question about preparing osmium tetroxide } } While we are on the topic of making up fixatives I have a } supplementary question. } } How do you open glass osmium tetroxide ampoules? } } I put mine in a 100ml glass bottle and then break it with a } glass rod. This is nerve-wracking, tedious (they do not } break readily) and not as safe a procedure as I would like. } } The ampoules used to have a weakened neck which could be } sawn off but they do not come in this form from our } supplier any more. } } Is there an easier way? } } Dave } } } On Thu, 6 Sep 2001 15:46:24 -0400 "Monson, Frederick C." } {fmonson-at-wcupa.edu} wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } I dissolve 1g OsO4 in 25ml double distilled water and store it in a } bottle } } with a Teflon lined screw cap. The bottle is then held in the } refrigerator } } in a small plastic jar (available from all lab distributors - mine from } } Fisher or S/P) with a screw cap . In this way, the plastic becomes } } osmicated NOT the refrigerator. I have done this for years with no } untoward } } results. } } } } I fix with aldehydes in screw cap scintillation vials (20ml or 5ml) } glass or } } plastic. Osmication is done in glass vials with Teflon lined screw caps } } using a minimum of OsO4 to do the job. } } } } I make glutaraldehyde from a 10ml vial in dd HOH (50%, 10ml Glu + 40ml } dd } } HOH yields 10% Glu) as a 10% solution. This in a screw top bottle with } a } } Teflon lined cap. I then make up an appropriate amount of fixative } using } } the 10% glu and 10X buffer concentrate where possible. The 10% Glu is } kept } } in a plastic jar in the refrigerator. } } } } I find that double containers are safe and reduce contamination } } considerably. } } } } Hope this helps. } } } } Fred Monson } } } } Frederick C. Monson, PhD } } Center for Advanced Scientific Imaging(CASI) } } West Chester University of Pennsylvania } } Schmucker Science Center II } } South Church Street } } West Chester, PA, 19383 } } eMail: fmonson-at-wcupa.edu } } http://darwin.wcupa.edu/casi/ } } } } } } ---------------------------------------- } Patton, David } Email: David.Patton-at-uwe.ac.uk } "University of the West of England" } }
Depends also from the measurement direction. The formula is right in the tilt direction, but the real tilt angle of a straight line on flat sample varies from nominal tilt angle in tilt direction to zero in the perpendicular direction. In the other directions, the formula is right too, but you must estimate the real tilt angle in the measurement direction, in respect to the tilt direction.
And of course, don't forget than in a SEM the perspective is not "conique" (I don't know the right terms in english) but "cavalière" . A tilted square looks like a rectangle and not a trapezoid. Try with a TEM grid with square holes, and try too with a grid with hexagonal holes. And rotate it ... You may feel dizzy, with high tilt angles. Our brain is not build for such a perspective ! But it can give nice pictures.
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess 67037 Strasbourg CEDEX France
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Depends upon the geometry of the sample: try making the calculations for a } sphere at a 45 degree angle. } } My guess is that the measurements would be the same as if the sphere were at } 0 degrees or 90 degrees. } } For purely planar sample the equations would be valid. } } regards, } } Earl } } ----- Original Message ----- } } From: "Nancy Zjaba" {nzjaba-at-alfalight.com} } To: "'microscopy-at-msa.microscopy.com'" {microscopy-at-sparc5.microscopy.com} } Sent: Thursday, September 06, 2001 9:18 AM } Subject: SEM: Tilt correction factor } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Hello to the list. } } } } It's been a while since I've been on the list, and since I've been working } } as a microscopist. } } } } If I image a sample in the SEM with the sample tilted, then want to make } } measurements from the images, what is the equation to use to correct for } the } } tilt? I know it involves the cosine of the tilt angle, but beyond that, } I'm } } drawing a blank. } } } } Thanks for your assistance. } } } } Nancy Zjaba, Technical Staff } } Alfalight Inc. } } 1832 Wright Street } } Madison, WI 53704 } } (608) 240-4875 } } nzjaba-at-alfalight.com } } } } } } } }
The method we use, which I teach to all the students, is to wrap the vial with a short length of paper towel (usually about four or five turns around the bottle), grasp the vial at both ends, and gently exert pressure with the thumbs. Then carefully unwrap the package and throw both vial halves in warm distilled water. I've never had any problem, either with crystals falling out in the paper or with glass poking through the paper towels. I was also taught the whack method, and found that my aim was not adequate to the job.
We use the same wrapping method with glutaraldehyde ampoules, but instruct the students to be sure to keep them upright.
Marie } } While we are on the topic of making up fixatives I have a } supplementary question. } } How do you open glass osmium tetroxide ampoules?
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-2131 University of Connecticut Storrs, CT 06269-2131 Phone: 860-486-3588 Fax: 860-486-6369
Regarding tilt correction, I enclose this from an earlier reply: it is a warning against the use of the tlit correction control on the SEM.
*** one thing is that [certain] morphology is generally quite low-profile and vague under SEM, _unless_ you tilt the specimen: I generally tilt at 54^ = ARC COS (1 / SQRT(3)), the "magic" angle which ensures that x, y, and z come out isometric, with the axes at 120^ to each other: if there is any orientation (say in an injection moulding or drawn specimen, then while the specimen is still flat-on, make sure that x and y are both at 45^ to the tilt axis. Then you will get isometric projection of all three axes. But NEVER use the tilt-correction control, or you will get distortion: for example, it makes spheres (which should appear circular at any angle) come out as ellipsoids.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
I have a diamond pen or scribe (bought mine from EMS) that I normally use for indelibly etching numbers on glass slides. I use it to carefully scratch a circular line completely around the narrowmost point of the ampoule. I usually go around several times to ensure a good scratch. I rinse the vial off with 95% ethanol and place inside a sturdy glass bottle (I use a Schott-brand glass bottle with orange plastic cap - available from major supply houses like fischer). I then shake the bottle vigorously (inside the fume hood, while silently praying it doesn't break!) It generally breaks within a few good snaps of the wrist. I add 50 mls of water and then store this bottle inside a plastic jar in my fume hood. The inside top of the plastic cap of the Schott bottle and the inside of the plastic jar go black with time and I periodically (yearly) replace them. I would never trust putting this stuff in my refrigerator. It leaks out the first bottle - why would one think it doesn't then leak out the second? I use it up usually within 3-4 months and never had a problem with room temperature storage over the last 20 years. If you use ampoules with pre-scored lines (easy snap open style) there is another trick to consider i used to use in the old days but it comes with a potential hazard that safety officers and lawyers advise against so I won't recommend it. Place about one cm of liquid nitrogen in a 50 ml disposable beaker in the hood. Slowly lower the ampoule into the liquid nitrogen. this causes the osmium to contract and break free from the glass. Snap the ampoule and pour the contents into water. the low temp also reduces fumes during pouring. this method avoids glass in your stock solution. the danger is that if there is a crack in your vial, the nitrogen would enter, and when warmed, potentially explode the vial. I never tried dry ice but i suspect it would work and avoid this hazard.
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-- Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
I have done CPD for years, and never seen this problem with EtOH dehydration (or acetone). The only times I've gotten anything like this is from the specimen container used for drying, or salt precipitation when the buffer was incompletely washed off. Another possibility is the CO2. Many CO2 tanks are full of water and crud. Is there a molecular sieve filter (for the water) and a membrane filter (for the crud) on the line from the tank to the CPD?
Mind, it contaminant *could* be from the EtOH, but that's just a reason to use better EtOH, not to switch to acetone.
Phil
} Hi, } } Recently, a collegue was having problems with a white crystaline } substance covering his samples of ear tissue after they were removed } from the critical point dryer. It was believed to be from a } contaminant in the ethanol and it was recommended to switch to acetone } dehydration as it was less likely to have this problem. I haven't done } SEM in years, so I haven't been paying attention to anything like this } on this listserve. I was wondering if other people experiencing this } problem. If you are using acetone, have you ever seen this? } } Thanks for your comments, } } Karen Pawlowski, Ph.D.
-- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
Hello all, I was wondering what computer programs people are using for 3-D reconstruction of TEM serial sections. I am interested in peoples opinions of the software (i.e. ease of use, learning curve, cross-platform functionality, cost, etc.). Feel free to email me off list if you choose. Also vendors feel free to send me any info you might have on the products you sell.
Thanks to all,
-EK
Eugene W. A. Krueger Sr. Research Technologist in Cell Biology II Center for Basic Research in Digestive Diseases Mayo Clinic and Foundation (507)284-0580 (lab) (507)284-0762 (fax) krueger.eugene-at-mayo.edu
see our web page at http://www.mayo.edu/mcniven_lab/
Another source that we have come across is sodium phosphate buffer crystals. If fixation is carried out with sodium phosphate buffered soluitons, and then immediately transittioned to dehydration solvent (either EtOH or Acetone) then we have seen samples coated with fine crystals. We use either serveral water rinses between fixation and dehydration or an alternative buffer (i.e. Na Cacodylate)
} } Recently, a collegue was having problems with a white crystaline } substance covering his samples of ear tissue after they were removed } from the critical point dryer. It was believed to be from a } contaminant in the ethanol and it was recommended to switch to acetone } dehydration as it was less likely to have this problem. I haven't done } SEM in years, so I haven't been paying attention to anything like this } on this listserve. I was wondering if other people experiencing this } problem. If you are using acetone, have you ever seen this? } } Thanks for your comments, } } Karen Pawlowski, Ph.D. } }
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
I would be interested in hearing from people anywhere in the world who have had PERSONAL experiences with the installation of electron microscopes or other sensitive equipment within 800 meters of electrically powered light rail, trains, buses or subway systems which have created vibrations or stray field problems for these instruments. I am specifically interested in hearing from those who have had prior experiences with light rail or bus transportation units powered by electricity. I would like to hear positive (no known problems for sensitive instruments) as well as negative experiences you have had or are having.
If you are aware that one or more of these types of transportation systems is within the 800 meter range of your equipment and specific measures were taken to eliminate vibration or stray fields, I will like to know that too.
You may wish to respond to me personally. Thank you.
John C. Wheatley Lab Manager Arizona State University Center for Solid State Science PSA-213 BOX 871704 Tempe, AZ 85287-1704
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Dave, I personally would not use a supplier who does not pack Osmium in easy open vials or ampoules such as the ones which have the thin neck that can be snapped open. It is just too dangerous to try to break the vials as you described.
Often the osmium crystals will adhere tightly to the glass. If you dip the vial into liquid nitrogen while holding it by the neck with forceps, the crystals will release from the glass surface. They can then be shaken down to one end of the vial.
You can purchase silicon ampoule breakers from some of the major EM supply houses. They are flexible molded silicon with a depression the shape ofthe ampoule. Just put the ampoule in, bend the mold and the ampoule will break. Then remove the portion with the crystals and pore them into your osmium container. You end up with the two large piece of glass which should be disposed of carefully since there might be traces of osmium remaining. The osmium crystals will normally dissolve into water (or buffer) if left overnight at room temperature....in a hood.
I normally make osmium up double strength in water and it will keep for months in the refrigerator, remaining a clear light amber solution. I also recommend double bottling to help prevent escape of vapors. However, we do not store any biologicals in the same refrigerator with fixatives. Regardless of how careful you try to be, the odds are that some fixative vapors will escape.
Debby
Debby Sherman Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-099e Whistler Building West Lafayette, IN 47907
--------------------------------------
While we are on the topic of making up fixatives I have a supplementary question.
How do you open glass osmium tetroxide ampoules?
I put mine in a 100ml glass bottle and then break it with a glass rod. This is nerve-wracking, tedious (they do not break readily) and not as safe a procedure as I would like.
The ampoules used to have a weakened neck which could be sawn off but they do not come in this form from our supplier any more.
Is there an easier way?
Dave
On Thu, 6 Sep 2001 15:46:24 -0400 "Monson, Frederick C." {fmonson-at-wcupa.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I dissolve 1g OsO4 in 25ml double distilled water and store it in a bottle } with a Teflon lined screw cap. The bottle is then held in the refrigerator } in a small plastic jar (available from all lab distributors - mine from } Fisher or S/P) with a screw cap . In this way, the plastic becomes } osmicated NOT the refrigerator. I have done this for years with no untoward } results. } } I fix with aldehydes in screw cap scintillation vials (20ml or 5ml) glass or } plastic. Osmication is done in glass vials with Teflon lined screw caps } using a minimum of OsO4 to do the job. } } I make glutaraldehyde from a 10ml vial in dd HOH (50%, 10ml Glu + 40ml dd } HOH yields 10% Glu) as a 10% solution. This in a screw top bottle with a } Teflon lined cap. I then make up an appropriate amount of fixative using } the 10% glu and 10X buffer concentrate where possible. The 10% Glu is kept } in a plastic jar in the refrigerator. } } I find that double containers are safe and reduce contamination } considerably. } } Hope this helps. } } Fred Monson } } Frederick C. Monson, PhD } Center for Advanced Scientific Imaging(CASI) } West Chester University of Pennsylvania } Schmucker Science Center II } South Church Street } West Chester, PA, 19383 } eMail: fmonson-at-wcupa.edu } http://darwin.wcupa.edu/casi/ } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Gray, Peter, 1975, Microtomist's Formulary and Guide, R. Krieger, Huntington, NY. (Still available for the hardy bibliophile at http://WWW.WEB4U.COM/cgi-bin/full_page?0-88275-247-2) reports FOUR fixative mixtures that are attributed directly to Champy:
My favorite, of course, is number 2!!! whose last component provides a wonderful bit of chemical history. We see how Bayer must have started. Dupont.
Regards, and I know that this doesn't help at all!
But it is all part of the art.
Fred Monson
} ---------- } From: Richard Edelmann } Reply To: edelmare-at-muohio.edu } Sent: Wednesday, September 5, 2001 4:24 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Bio. Fixation: What is Champy's Fluid? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } "Fixed 2-5 minutes in Champy's fluid, rinsed 2x in water" . . . followed } by } ETOH dehydration, CPD, mounting and SEM viewing. } } } O.k., we give up, searched numberous books, papers, catalogs, web } sites, etc. . . . does anyone have a recipe for "Chmapy's fluid" a.k.a. } "Champy's Solution"? Please? } } Thank you much! } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 352 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } } "RAM disk is NOT an installation procedure." } }
EMS sells an ampoule breaker , Cat# 60600 for $12. We've used them for breaking ampoules of paraformaldehyde, glutaraldehyde, acrolein and osmium within seconds. The soft silicon molds can be twisted to achieve a quick break. I usually tap the osmium tetroxide crystals to the bottom and use forceps to lift the open ampoule into my prepared 50 ml of distilled water to make our stock 2%. I have no financial interest in this company but am a satisfied customer for many years. Phone number is: (800) 523-5874. Rosemary Walsh, EMF for the Life Sciences PSU
Fred: Have you given any though to the fact that Coriolis' forces are different in the southern hemispher? Water drains from a basin in a counterclockwise motion south of the Equator and clockwise north of the Equator. Maybe wiggles and waggles are affected thesame way between the eastern and western hemispheres.
Regards,
Sam Purdy
Technical Center National Steel Corp. 1745 Fritz Dr. Trenton MI Voice:734-676-2682 FAX: 734-676-2682 spurdy-at-nationalsteel.com
Use a triangular metal file to score around the neck, then use the usual precautions (protect fingers, open in hood , etc.) to pop open the vial. The EMS brand I use breaks open easily by wrapping paper towel/ gauze around top and gentle application of pressure.
Becky Garrison I am not an employee of any EM supplier.
-----Original Message----- } From: Patton, David [mailto:David.Patton-at-uwe.ac.uk] Sent: Friday, September 07, 2001 5:31 AM To: Monson, Frederick C. Cc: 'Microscopy Listserver'
While we are on the topic of making up fixatives I have a supplementary question.
How do you open glass osmium tetroxide ampoules?
I put mine in a 100ml glass bottle and then break it with a glass rod. This is nerve-wracking, tedious (they do not break readily) and not as safe a procedure as I would like.
The ampoules used to have a weakened neck which could be sawn off but they do not come in this form from our supplier any more.
Is there an easier way?
Dave
On Thu, 6 Sep 2001 15:46:24 -0400 "Monson, Frederick C." {fmonson-at-wcupa.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I dissolve 1g OsO4 in 25ml double distilled water and store it in a bottle } with a Teflon lined screw cap. The bottle is then held in the refrigerator } in a small plastic jar (available from all lab distributors - mine from } Fisher or S/P) with a screw cap . In this way, the plastic becomes } osmicated NOT the refrigerator. I have done this for years with no untoward } results. } } I fix with aldehydes in screw cap scintillation vials (20ml or 5ml) glass or } plastic. Osmication is done in glass vials with Teflon lined screw caps } using a minimum of OsO4 to do the job. } } I make glutaraldehyde from a 10ml vial in dd HOH (50%, 10ml Glu + 40ml dd } HOH yields 10% Glu) as a 10% solution. This in a screw top bottle with a } Teflon lined cap. I then make up an appropriate amount of fixative using } the 10% glu and 10X buffer concentrate where possible. The 10% Glu is kept } in a plastic jar in the refrigerator. } } I find that double containers are safe and reduce contamination } considerably. } } Hope this helps. } } Fred Monson } } Frederick C. Monson, PhD } Center for Advanced Scientific Imaging(CASI) } West Chester University of Pennsylvania } Schmucker Science Center II } South Church Street } West Chester, PA, 19383 } eMail: fmonson-at-wcupa.edu } http://darwin.wcupa.edu/casi/ } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
David Patton wrote: ======================================================== While we are on the topic of making up fixatives I have a supplementary question.
How do you open glass osmium tetroxide ampoules?
I put mine in a 100ml glass bottle and then break it with a glass rod. This is nerve-wracking, tedious (they do not break readily) and not as safe a procedure as I would like.
The ampoules used to have a weakened neck which could be sawn off but they do not come in this form from our supplier any more.
Is there an easier way? ========================================================== I am a bit surprised to hear that you are getting osmium tetroxide that is not in prescored ampoules. So far as I know, all of the major suppliers of EM chemicals, like SPI, have long since supplied their ampoules in the pre- scored form. The only time I have seen in recent years ampoules not in pre- scored form are those that are destined for other markets and applications.
Also, SPI, as well as the other major suppliers have long offered ampoule neck breakers, such as are described on URL http://www.2spi.com/catalog/tools/smtol21.html These are meant to be used one time and discarded. They are inexpensive and seem to provide an extra element of safety.
Using only prescored ampoules and an ampoule neck breaker might be easier on the nerves.....
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
Below are the two answers to your question, since I am not sure whether you have obj. aperture inserted in the beam, or you don't. Both assuming that the field of view that you mentioning is the image of the sample in the holder, rather than an illumination circle.
1. If obj. apertures holder is inserted in the beam. Objective aperture should not be used in LM range. In Philips TEMs starting with EM400 series , the main optical configuration difference between LM and M modes is very distinct and as followed: in LM mode, the objective lens current is very low (few hundreds milliAmps, depending on HT set), and is fixed- hence magnification of obj. lens is also low (between 1 and 10, closer to 1- just a guess). Focusing in LM mode is done by diffraction lens. Almost all magnification is accomplished by lenses below the objective lens. Obj. aperture is located between the specimen and that lens system, much closer to the specimen, than to the first magnifying (diffraction) lens, and is blocking most of the image if inserted in the beam.
In M mode, the obj. lens current is much higher (several Amps, depending on the particular TEM model and the HT set), making magnification of the obj. lens in M mode much higher (say, 50 or so). You can use obj. aperture now, because of the significant part of the magnification happens very close to the obj. aperture, plus much smaller area of the specimen is forming first (objective lens) image. Focusing in M mode is done by the obj. lens. Diffraction lens current in M mode is fixed and depends on the HT and magnification set.
This two modes are "toggled" when TEM magnification is switched LM to M and vice versa, in a single magnification step.
2. If obj. apertures holder is withdrawn from the beam. When the condenser aperture is out of the beam, field of view in the lower LM range increases. Nevertheless, obj. aperture holder shall not be visible when withdrawn from the beam (control lever to the right). If obj. apertures holder is still visible, then either the column alignment, or the obj. aperture mechanical alignment (or both...) are incorrect.
If you are referring to illumination circle with the specimen holder withdrawn- I wouldn't care much about obj. aperture holder visible on the periphery of the illumination circle at the lowest LM steps, as long as it is not blocking the sample image at the corresponding extreme of the sample travel, and as long as everything else looks normal.
If you need to align the "out" position of any aperture holder on this TEM, turn little Allen set screw located on the right side of the appropriate aperture holder mechanism housing.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax
This message is made of 100% recycled electrons. -----Original Message----- } From: "marienti-at-tiscalinet.it"-at-sparc5.microscopy.com {"marienti-at-tiscalinet.it"-at-sparc5.microscopy.com} To: microscopy-at-sparc5.microscopy.com {microscopy-at-sparc5.microscopy.com}
How would you like to escape the northern winter this year?
Yes? Then join us for the 17th Australian Conference on Electron Microscopy, to be held in Adelaide, South Australia, from February 4 - 8, 2002. Encompassing the areas of Electron, Scanned Probe and Confocal Microscopies, this important meeting will provide an opportunity to explore the many applications of these techniques, to keep abreast of the latest developments and to meet and mix with many of Australia's and the world's eminent leaders in the field. This meeting was last held in Adelaide in 1988. A lot has changed in this city in the past 14 years. Adelaide has emerged as a vital city which can offer the visitor a range of exciting opportunities. The local wine and food have attained international recognition. Tourists are welcomed and well catered for. The conference will be held in the brand new Adelaide Convention Centre which is conveniently located in the CBD, close to accommodation and other facilities. International visitors should take some extra time either before or after the conference to explore Adelaide and South Australia. Tours can be arranged to take you to the sights of Adelaide, the nearby picturesque Adelaide Hills, and the wine growing regions of McLaren Vale, the Barossa Valley and Clare Valley. Each of these areas has its own unique history, heritage sites and gourmet delights! Adelaide's clean white beaches are within easy reach of the city. Experience the wildlife, flora and landscape of Kangaroo Island. Adelaide is also the perfect stepping-off point to locations such as the beautiful Flinders Ranges, where centuries-old Aboriginal art and ancient geological formations can be seen, Uluru, the opal mining town of Coober Pedy, and the Outback. The current exchange rates ensure that travel in Australia has never been more economical for the overseas tourist. A series of pre-conference workshops will be held to allow participants to gain hands-on experience in the latest techniques and applications. Many of the keynote speakers will participate as presenters, providing a wonderful opportunity for interaction with these noted scientists. Be sure to book early, as places in some of the workshops is limited. During the week, delegates will have the chance to mingle and relax after hours in several of Adelaide's noted heritage locations. A range of accommodation choices, from five-star to budget, ensures choices suitable for all. With the deadline for abstract submission fast approaching, (October 6), start making plans now to join us in February. Abstracts should be submitted electronically, and the registration form can be downloaded from the website. Full details can be obtained at http://www.adelaide.edu.au/CEMMSA/acem17 Keep an eye on the website for regular updates. We look forward to seeing you at ACEM17.
ACEM17 Organising Committee
CEMMSA Centre for Electron Microscopy and Microstructure Analysis Adelaide University Adelaide SA 5005 Ph: (08) 8303 4074 Fax: (08) 8303 4356 Website: http://www.adelaide.edu.au/CEMMSA
At 10:30 7/09/01 +0100, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
we 1. thouroughly clean off any label and label glue from the vial
2. put a scratch right around the vial
3. THEN it becomes easier to smash in the bottle, as you do it.
AND if you want, you can promote a crack around the line of the scratch by touching a redhot glass rod to the scratch.
AND if you want, you can distribute the OsO4 in a thinner layer around the wall of the vial by warming the (unscratched) vial in hot water, then rolling the vial as the OsO4 cools.
This accelerates solution.......
AND if your OsO4 starts to go dark, you can refresh it almost completely by adding hydrogen peroxide dropwise and thus titrate it to a clear solution again. Oten you can restore it to straw colour, but we will use even a pale purple solution as it will have enough tetroxide in it to work.
Upon reading subsequent posts about breaking OsO4 vials, I realized I must amend my suggestion of wrapping the neck with paper towels and gently applying pressure to break at the neck. This method works fine on PRE-SCORED vials, but almost certainly is unsafe or ineffective when used on unscored vials. I wasn't aware that unscored vials were still being sold.
Marie
Dr. Marie E. Cantino Dept. of Physiology and Neurobiology, U-2131 University of Connecticut Storrs, CT 06269-2131 Phone: 860-486-3588 Fax: 860-486-6369
I have an Hitachi H7100FA TEM on the second floor of a building built almost directly over an underground electric railway. When I moved in, bringing the EM with me, there was the choice of various locations. I spent very little time standing on a wooden floor feeling it shake under me. I spent considerably more time sitting on a concrete floor with a saucer of water watching for vibrations (we had no money to pay experts). Apart from having someone ask me if I needed medical help, I was left in peace with my saucer and am now quite happy with the chosen location. I don't work at REALLY high mags, but the resolution test was accomplished without any vibrations and I've coexisted happily with the trains for 3 years. Before that I was on the 2nd floor of a concrete building next to a busy road and it was dreadful. The building construction and location within it was the important aspect for me.
Diana
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Diana van Driel Department of Clinical Ophthalmology Sydney University GPO Box 4337 Sydney NSW AUSTRALIA 2001
We are currently running an Oxford ISIS EDS system and an Oxford MonoCL cathodoluminescence system from the same computer and on the same SEM. We want to split the Cathodoluminescence system off onto another column so we need another ISIS board enclosure (box).
We would be pleased to hear from anyone upgrading their system who would have an ISIS box that is spare. (The box that houses the ISIS boards, not the PC.) We do not specially need the ISIS boards.
We will pay a reasonable amount (say US$5,000) plus shipping for an ISIS box in working order.
Dr. Mel Dickson, Deputy Director, The Electron Microscope Unit, Adjunct Associate Professor, School of Microbiology & Immunology The University of New South Wales UNSW SYDNEY 2052 Australia. Phone +612 9385 6383 Fax +612 9385 6400 http://srv.emunit.unsw.edu.au/
I'd like to prepare catalyst particles for TEM viewing by ultramicrotomy. The problem is that I want to study the location and nature of carbon in these particles and I am not certain how to distinguish the carbon from the epoxy. We currently use Spurr's resin, but is there an epoxy I can use that is not carbon based ?
TIA Willem Erasmus
Willem Erasmus Snr. Scientist, Basic Catalysis Research Sasol Technology Tel : +27 +16 960 - 4211/2772 Fax : +27 +16 960 - 2826 E-mail : willem.erasmus-at-sasol.com PO Box 1, Sasolburg, 1947, Republic of South Africa
[All views expressed are my own and not necessarily that of my employer.]
Could be interesting for you... ----- Original Message ----- } From: "Steven Dessein" {steven.dessein-at-BIO.KULEUVEN.AC.BE} Dear Taxacomers,
The Laboratory of Plant Systematics (K.U.Leuven, Belgium) today released Carnoy. Carnoy is an easy-to-use image analysis program, designed to carry out measurements on digital images and especially on micrographs obtained from light or electron microscopes.
Carnoy features a unique one-click calibration option: click on the scale bar, enter its length and you are ready to carry out real-world measurements on your micrographs. Carnoy measures length and surface area of regular and irregular objects. It also performs automated particle analysis (counting and measuring). The program can handle BMP, PICT, Photoshop, JPEG, GIF, PNG, SGI, TGA, TIFF and QuickTime image files and can convert between these file types. Measurements are stored in a list that smartly manages all data. Every measurement can be annotated. Measurements can be exported to a tab-delimited file, which can be read by any text editor, word processor, database or spreadsheet, for further analysis. The program's clean and easy-to-use interface enables you to be productive within minutes.
Carnoy can be used in the following fields: taxonomy, morphology, ecology, physiology, molecular cell biology, agricultural sciences, paleontology, archaeology, among others. Carnoy requires a Macintosh running Mac OS X or a PC with at least 64 MB RAM and Windows 95 / 98 / ME / NT / 2000 / XP. A resolution of 1024 x 768 pixels (or higher) is recommended.
Carnoy is $15 shareware (if you cannot afford this, please, contact peter.schols-at-bio.kuleuven.ac.be). It is available for download on the Carnoy website. http://www.carnoy.org
Kind regards,
P. Schols, S. Dessein & E. Smets
Steven Dessein Laboratory of Plant Systematics Institute of Botany and Microbiology Kardinaal Mercierlaan 92 B-3001 Leuven Belgium
I'm not sure if our Osmium vials are prescored or not, but we've never had any problems or the nerve wracking experiences that you have mentioned. We score around the vial a couple of times with a diamond pen(made especially for marking on glass). The side where we want the break is scored a couple times more. In the fume hood we wrap the vial in a paper towel and snap it facing toward the back of the hood. Hope this helps.
Paula Moore Wake Forest University School of Medicine Pathology/EM Lab pmoore-at-wfubmc.edu
} } How do you open glass osmium tetroxide ampoules? } } I put mine in a 100ml glass bottle and then break it with a glass rod. } This is nerve-wracking, tedious (they do not break readily) and not as safe } a procedure as I would like. } } The ampoules used to have a weakened neck which could be sawn off but they } do not come in this form from our supplier any more. } } Is there an easier way? } ========================================================== } } } Using only prescored ampoules and an ampoule neck breaker might be easier on } the nerves..... } } Chuck } } ==================================================
Amray 1610 SEM with PGT EDS free to a good home---interested party pays all shipping charges. Please send queries off-list. Thanks!
Melissa Troost Electron Probe Instrumentation Center Northwestern University Evanston, Illinois, USA 847-491-3439 m-troost-at-northwestern.edu
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Does anyone have or know of a source of SEM photomicrographs of bacteria, mold and yeast. I am asking on behalf of a colleague who will pay a royalty for the use of the photos.
Mark Germani, Ph.D. MicroMaterials Research, Inc. 136 Shore Drive, #200 Burr Ridge, IL 60521
Anyone out there knows how to run up an X-ray film (exposed as well as unexposed) for TEM?????
Dear Neelima, What do you mean by "run up"? We are lucky to have our LoDose and MRF32 x-ray films cut to the appropriate size--6.5 x 9 cm--and we treat them like other EM film EXCEPT, we load and develop them in total darkness, and we use x-ray film developer in the tank instead of D-19. Our darkroom is in the same room as the scope, so transport can occur in total darkness. If this is not the case for your facility, and you have to run the film up to your darkroom, get a film transport bag from a photo supply store, put the film in its original box, put the box in the bag, and carry it to your darkroom. The bags are designed to be light tight. Good luck, and if you mean something else by "run up", let the list know, and I'll try to give you a better answer. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
Karen: Stick to absolute ethyl alcohol (Et0H). Never use acetone with any critical point drying apparatus that have glass viewing windows held in place with a epoxy resin that would be attacked by acetone. The white crystals may very well be buffer salts. You can start dehydrating with a larger ratio of water to ethyl alcohol and let stand for a longer time to remove buffer salts while working up to absolute strength ethanol. For some samples such as tissue culture monolayers the critical point dryers can be avoided altogether by proceeding from the absolute ethyl alcohol to 1,1,1,3,3,3-hexamethyldisilazane to air dry. I would recommend the 99.9% pure solution.
Henry
Henry Eichelberger, Manager Electron Microscopy Facility Department of Biological Sciences State University of New York-Binghamton Binghamton, NY 13902-6000
} ---Original Message------------------ } Recently, a collegue was having problems with a white crystaline } substance covering his samples of ear tissue after they were removed } from the critical point dryer. It was believed to be from a } contaminant in the ethanol and it was recommended to switch to acetone } dehydration as it was less likely to have this problem. I haven't done } SEM in years, so I haven't been paying attention to anything like this } on this listserve. I was wondering if other people experiencing this } problem. If you are using acetone, have you ever seen this? } } Thanks for your comments, } } Karen Pawlowski, Ph.D. } }
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
I got a problem when preparing TEM foils of Al-alloy by means of electro-polishing. A dark film produced on the specimen surface when it was been polishing. I think, the polishing condition is right and listed below:
Solution, 25 % Nitric acid plus 75 % Mechenol Temperature, about -30 degree C Voltage, 15-20V
I would be pleased to hear from anyone who has an idea to sort out this problem.
Dear Randy, Drosophila brains are very hard to fix. The neurons are very delicate and lose their structure when not well preserved. What age are the bugs? You might try some adding some acrolein (we used 1% acrolein and 2.5% glutaraldehyde in 0.1 M cacodylate buffer, pH 7.1) and leave out the paraformaldehyde. See if you still have immuno-reactivity You don't need to use propylene oxide. You can use 100% EtOH instead. It is miscible with resins, as long as it is totally dry. Also, don't store your specimens in buffer, but process straight through. I do microwaving these days, so time is never an issue anymore. Good luck!
JoAnn Buchanan Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94305 650-723-5856
04:18 PM 09/06/2001 -0500, you wrote:
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In conjunction with the Pacific Northwest Microscopy Society, The University of Washington Core for Communication Research, Center on Human Development and Disability and the W. M. Keck Center for Advanced Studies in Neural Signaling present:
Microscopy and Digital Imaging: Advances & Applications
September 27, 2001 12:30 - 5 p.m. Location: CD 150
12:30 "Multiphoton and Multispectral Imaging in Vertebrates" Mary Dickinson, Caltech
1:15 "Some New Approaches to Intracellular Imaging" Bob Fern, University of Washington
2:00 FRET imaging with the confocal microscope Kolja Wawrowsky, Leica
2:45 Break
3:15 "In Vivo Microscopy of Murine Brain" Liz Yoder, UCSD
4:00 "Deconvolution Methods in Wide-field and Confocal Microscopy" Paul Goodwin, Applied Precision Inc
5:00 Reception
The Veteran's Administration Hospital and Medical Center Morphological Analysis Core Facility will have an open house and talks on September 28, 2001.
Speakers sponsored by Applied Precision, BioRad, Leica, and Zeiss The reception is sponsored by Bartels and Stout, Inc.
Contact Paulette Brunner pbrunner-at-u.washington.edu or Glen MacDonald glenmac-at-u.washington.edu for information.
Regards, Glen -- Glen MacDonald Microscopy and Imaging Facility University of Washington Core for Communication Research Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 glenmac-at-u.washington.edu (206) 616-4156 (206) 616-1828 fax **************************************************************************
C:} The box said "Requires Windows95 or better". So I bought a Macintosh. **************************************************************************
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sbcook-at-bcpl.net) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, September 9, 2001 at 22:52:34 ---------------------------------------------------------------------------
Email: sbcook-at-bcpl.net Name: Katelyn Cook
Organization: St. Michael the Archangel
Education: 6-8th Grade Middle School
Location: Baltimore, MD
Question: I am planning to do a science fair project about germs and the importance of hand washing. I want to collect germs from common public places such as doorknobs, handrails, telephones, sink faucets, etc. What is the best way to collect the germs? Will I need to grow the germs in a petri dish after I collect them? Will I be able to identify different types of germs using my microscope?
I have seen cellular voids from poor fixation as well as from genetic mutations. The biggest impediment I have when fixing drosophila brains is getting the fixative to the tissue. I recommend cutting the neck as well as the proboscis and then using gentle centrifugation (microcentrifuge ~2000rpm for one minute) to get the heads to sink. Due to air in the brain cavity this last step does not always work. If you use 2% glutaraldehyde and osmium post fixation do you get good ultrastructure? In a project I did some time ago I had the best results using 0.07 M sodium cacodylate buffer at pH 7.5. That gives an osmolarity of about 187mOsm. However, I routinely use 0.1 M sodium cacodylate and find that adequate.
At 04:18 PM 9/6/01 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Liststers; } } I'm working with SEM of pre-colonial potteries from Pantanal at the = } Brazilian West border with Bolivia. In some samples we've found shell = } and sponge spicule in the bulk of potteries. We would appreciate any = } article or book with micrographies of similar materials in pottery ? = } Also, if someone could provide me references on clay with sponge = } spicule, would be very helpful. } } Ubirajara Pereira Rodrigues-Filho Instituto de Química de São Carlos Universidade de São Paulo São Carlos, Brazil
Cameca MBX Microprobe is available at this time. 4 wavelength spectrometers (includes PC2 crystal for Lt ele. spec.) Kevex Sigma EDS New GW Backscattered system PC Based operating system Diff pump vacuum system Cathodoluminescence system (OEM) System is about 17 years old and in good condition (at last service call). Has been turned off for 1 year.
Please send inquires to E-MAC Walter Protheroe corvos-at-aol.com
You may want to try the following website at Utah State University:
http://bioweb.usu.edu/emlab/Galleries.html
They have many great SEM images. A contact person there to talk to would be:
Bill McManus TEL: (435) 797-1920
I hope this helps.
Best regards-
David
Mark Germani wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Does anyone have or know of a source of SEM photomicrographs of } bacteria, mold and yeast. I am asking on behalf of a colleague who will } pay a royalty for the use of the photos. } } Mark Germani, Ph.D. } MicroMaterials Research, Inc. } 136 Shore Drive, #200 } Burr Ridge, IL 60521 } } (630) 325-8170 } (630) 325-8178 fax } } mgermani-at-micromaterialsresearch.com
-- David Henriks TEL: +1-949-492-2600 South Bay Technology, Inc. FAX: +1-949-492-1499 1120 Via Callejon San Clemente, CA 92673 USA e-mail: henriks-at-southbaytech.com
Dear Randy, Drosophila brains are very hard to fix. The neurons are very delicate and lose their structure when not well preserved. What age are the bugs? You might try some adding some acrolein (we used 1% acrolein and 2.5% glutaraldehyde in 0.1 M cacodylate buffer, pH 7.1) and leave out the paraformaldehyde. See if you still have immuno-reactivity You don't need to use propylene oxide. You can use 100% EtOH instead. It is miscible with resins, as long as it is totally dry. Also, don't store your specimens in buffer, but process straight through. I do microwaving these days, so time is never an issue anymore. Good luck!
JoAnn Buchanan Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94305 650-723-5856
04:18 PM 09/06/2001 -0500, you wrote:
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Hello everyone, I have a researcher here that is having some difficulty with getting good histology at the EM level. She is fixing Drosophlia brains with 4%para/0.5% glut in phosphate buffer pH7.2, no osmium post fix, ethanol dehaydration, propylene oxide as a transitional solvent to polybed 812, then performing post sectioning immunolabeling. The tissue has "holes" in it when viewed at the TEM. They are not holes in the section, but appear to be voids in the tissue. We have tried leaving out the propylene oxide, we have osmicated, and have looked at osmotic strength of the fixative (and many variations of the above, to no avail). Can anyone offer advice? Randy Nessler Univ. of Iowa
Yan, Assuming (sorry for that) that the polishing conditions are correct, and you are producing well dished samples with thin area and the only problem is sludge on the surface, try two things before changing any other conditions and try them one at a time so as not to confuse the issue. 1: Try increasing the jet speed. If this doesn't work. 2: Try increasing the temperature about 5 degs. It would be helpful to know what type of jet thinner you are using and exactly what the alloy is. Good Luck! Dorrance
} ---------- } From: yan.hu } Sent: Tuesday, September 11, 2001 12:35 AM } To: microscopy-at-sparc5.microscopy.com } Subject: ask help } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Listers, } } I got a problem when preparing TEM foils of Al-alloy by means of } electro-polishing. A dark film produced on the specimen surface when it } was been polishing. I think, the polishing condition is right and } listed below: } } Solution, 25 % Nitric acid plus 75 % Mechenol } Temperature, about -30 degree C } Voltage, 15-20V } } I would be pleased to hear from anyone who has an idea to sort out this } problem. } } } Regards } } Yan } ---------------------- } } yan.hu-at-brunel.ac.uk } } }
On Tue, 11 Sep 2001 05:22:19 -0500, sbcook-at-bcpl.net wrote:
| ------------------------------------------------------------------------ | The Microscopy ListServer -- Sponsor: The Microscopy Society of America | To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com | On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html | -----------------------------------------------------------------------. | | | Below is the result of your feedback form (NJZFM-ultra-55). It was | submitted by (sbcook-at-bcpl.net) from | http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, | September 9, 2001 at 22:52:34 | --------------------------------------------------------------------------- | | Email: sbcook-at-bcpl.net | Name: Katelyn Cook | | Organization: St. Michael the Archangel | | Education: 6-8th Grade Middle School | | Location: Baltimore, MD | | Question: I am planning to do a science fair project about germs and | the importance of hand washing. I want to collect germs from common | public places such as doorknobs, handrails, telephones, sink faucets, | etc. What is the best way to collect the germs? Will I need to grow | the germs in a petri dish after I collect them? Will I be able to | identify different types of germs using my microscope? | | --------------------------------------------------------------------------- |
Dear Katelyn Cook - While I was teaching as a graduate teaching assistant for a general biology class, I had the opportunity to lead a group of non- science college students on an examination of the microbial world around them. What I did was to have each of them use a sterile, cotton- tipped swab, moisten it a bit with sterile water (perhaps boiled water will do just as well), swab the surface of whatever object they wanted to explore, and then swab it onto the surface of a petri plate that already had bacterial medium. They then left all the plates for me to seal and incubate at room temperature for several days (or, at most, 1 week). When they got the plates back, they then could see with the naked eye the various colonies that formed and their various colors. If a microscope is available, then it is possible to identify individual bacteria by using a "wet mount" (that is, a slide with a small drop of sterile water and a very small amount of whatever colony is chosen). Although I don't have any experience with grade school students, I believe that my approach could work. So... to answer your questions: (1) A very easy way to collect bacteria is to first get some sterile swabs. The fact that they are sterile is important because any box of ordinary (or household) swabs is likely to already have bacteria on them, and obviously that would defeat the purpose of your experiment. As a result, you'll need some sterile swabs -- quite likely, they can be bought at a pharmacy near you when you look for dressings, bandages, etc. (2) Assuming you have sterile swabs, you could conceivably boil some water, let it cool to room temperature, and let each student lightly wet a swab (or more as needed). Why boil the water? For the same reason as needing sterile swabs -- because ordinary tap water will very likely contain bacteria (even if it looks perfectly clear). One alternative is to use distilled water if boiling isn't feasible. (3) Yes, you will need a petri plate -- the plate needs to have some kind of bacterial medium capable of supporting their growth; otherwise, your swabbing exercise will be fruitless. I do not know where such plates can be obtained, though, and you may want to seek others on this listserver; finally, (4) The use of a microscope is needed only if you're interested in examining individual cells. The typical colonies that are seen on the surface of a plate tend to vary in terms of both color and the type of colony produced. That is, a colony could be white or yellow, but its 'type" could be of a sticky nature or a more "liquid-like" nature. That part could be revealing on its own, or perhaps not. If you feel that a microscope is needed, then a simple way to see individual cells is to simply add a small drop of boiled / sterile water onto a slide, use an inoculating loop or needle, and gently pick up a small amount of a colony, and spread it around within to the drop. Place a coverslip over the slide, and put it onto the stage of a microscope. Try to use the highest objective lens magnification that is possible -- if there are a 10X, 20X and 100X objectives, use the 20X as the individual cells are very small in size -- and, if it's possible, try to set the microscope under non-brightfield light conditions such as phase-contrast. Bacterial cells are, generally, transparent to light, and as a result are often very hard to see because they have very little contrast with the background light. Anyway .. I hope you have some success with your experiment for these grade-school students. It is my my belief that microscopic, living things (such as the "germs" and, more specifically, bacteria, algae, etc.) are always fascinating things for kids to see, and I hope it turns out very well. Good luck ! Nelson Conti Former graduate student at San Francisco State University (in San Francisco, CA) A Bachelor's and Master's degree in Microbiology
_______________________________________________________ Send a cool gift with your E-Card http://www.bluemountain.com/giftcenter/
Have you contacted Oxford Instruments? With the INCA doing as well as they tell me it is they must be taking in quite a few ISIS towers on upgrades (they've had one of ours). Of course if you expect a full manufacturer's warrantee it might be pricey but if you took the same 'as seen' conditions that you would accept from another user then they might deal.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Listers, } } We are currently running an Oxford ISIS EDS system and an Oxford MonoCL } cathodoluminescence system from the same computer and on the same SEM. We } want to split the Cathodoluminescence system off onto another column so we } need another ISIS board enclosure (box). } } We would be pleased to hear from anyone upgrading their system who would } have an ISIS box that is spare. (The box that houses the ISIS boards, not } the PC.) We do not specially need the ISIS boards. } } We will pay a reasonable amount (say US$5,000) plus shipping for an ISIS } box in working order. } } } } } Dr. Mel Dickson, } Deputy Director, The Electron Microscope Unit, } Adjunct Associate Professor, School of Microbiology & Immunology } The University of New South Wales } UNSW SYDNEY 2052 } Australia. } Phone +612 9385 6383 Fax +612 9385 6400 } http://srv.emunit.unsw.edu.au/ }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
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EMS sells an ampoule breaker , Cat# 60600 for $12. We've used them for breaking ampoules of paraformaldehyde, glutaraldehyde, acrolein and osmium within seconds. The soft silicon molds can be twisted to achieve a quick break. I usually tap the osmium tetroxide crystals to the bottom and use forceps to lift the open ampoule into my prepared 50 ml of distilled water to make our stock 2%. I have no financial interest in this company but am a satisfied customer for many years. Phone number is: (800) 523-5874. Rosemary Walsh, EMF for the Life Sciences PSU
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-- [ From: Garber, Charles A. * EMC.Ver #3.1 ] --
David Patton wrote: ======================================================== While we are on the topic of making up fixatives I have a supplementary question.
How do you open glass osmium tetroxide ampoules?
I put mine in a 100ml glass bottle and then break it with a glass rod. This is nerve-wracking, tedious (they do not break readily) and not as safe a procedure as I would like.
The ampoules used to have a weakened neck which could be sawn off but they do not come in this form from our supplier any more.
Is there an easier way? ========================================================== I am a bit surprised to hear that you are getting osmium tetroxide that is not in prescored ampoules. So far as I know, all of the major suppliers of EM chemicals, like SPI, have long since supplied their ampoules in the pre- scored form. The only time I have seen in recent years ampoules not in pre- scored form are those that are destined for other markets and applications.
Also, SPI, as well as the other major suppliers have long offered ampoule neck breakers, such as are described on URL http://www.2spi.com/catalog/tools/smtol21.html These are meant to be used one time and discarded. They are inexpensive and seem to provide an extra element of safety.
Using only prescored ampoules and an ampoule neck breaker might be easier on the nerves.....
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
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Use a triangular metal file to score around the neck, then use the usual precautions (protect fingers, open in hood , etc.) to pop open the vial. The EMS brand I use breaks open easily by wrapping paper towel/ gauze around top and gentle application of pressure.
Becky Garrison I am not an employee of any EM supplier.
-----Original Message----- } From: Patton, David [mailto:David.Patton-at-uwe.ac.uk] Sent: Friday, September 07, 2001 5:31 AM To: Monson, Frederick C. Cc: 'Microscopy Listserver'
While we are on the topic of making up fixatives I have a supplementary question.
How do you open glass osmium tetroxide ampoules?
I put mine in a 100ml glass bottle and then break it with a glass rod. This is nerve-wracking, tedious (they do not break readily) and not as safe a procedure as I would like.
The ampoules used to have a weakened neck which could be sawn off but they do not come in this form from our supplier any more.
Is there an easier way?
Dave
On Thu, 6 Sep 2001 15:46:24 -0400 "Monson, Frederick C." {fmonson-at-wcupa.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I dissolve 1g OsO4 in 25ml double distilled water and store it in a bottle } with a Teflon lined screw cap. The bottle is then held in the refrigerator } in a small plastic jar (available from all lab distributors - mine from } Fisher or S/P) with a screw cap . In this way, the plastic becomes } osmicated NOT the refrigerator. I have done this for years with no untoward } results. } } I fix with aldehydes in screw cap scintillation vials (20ml or 5ml) glass or } plastic. Osmication is done in glass vials with Teflon lined screw caps } using a minimum of OsO4 to do the job. } } I make glutaraldehyde from a 10ml vial in dd HOH (50%, 10ml Glu + 40ml dd } HOH yields 10% Glu) as a 10% solution. This in a screw top bottle with a } Teflon lined cap. I then make up an appropriate amount of fixative using } the 10% glu and 10X buffer concentrate where possible. The 10% Glu is kept } in a plastic jar in the refrigerator. } } I find that double containers are safe and reduce contamination } considerably. } } Hope this helps. } } Fred Monson } } Frederick C. Monson, PhD } Center for Advanced Scientific Imaging(CASI) } West Chester University of Pennsylvania } Schmucker Science Center II } South Church Street } West Chester, PA, 19383 } eMail: fmonson-at-wcupa.edu } http://darwin.wcupa.edu/casi/ } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
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Fred: Have you given any though to the fact that Coriolis' forces are different in the southern hemispher? Water drains from a basin in a counterclockwise motion south of the Equator and clockwise north of the Equator. Maybe wiggles and waggles are affected thesame way between the eastern and western hemispheres.
Regards,
Sam Purdy
Technical Center National Steel Corp. 1745 Fritz Dr. Trenton MI Voice:734-676-2682 FAX: 734-676-2682 spurdy-at-nationalsteel.com
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Karen:
Another source that we have come across is sodium phosphate buffer crystals. If fixation is carried out with sodium phosphate buffered soluitons, and then immediately transittioned to dehydration solvent (either EtOH or Acetone) then we have seen samples coated with fine crystals. We use either serveral water rinses between fixation and dehydration or an alternative buffer (i.e. Na Cacodylate)
} } Recently, a collegue was having problems with a white crystaline } substance covering his samples of ear tissue after they were removed } from the critical point dryer. It was believed to be from a } contaminant in the ethanol and it was recommended to switch to acetone } dehydration as it was less likely to have this problem. I haven't done } SEM in years, so I haven't been paying attention to anything like this } on this listserve. I was wondering if other people experiencing this } problem. If you are using acetone, have you ever seen this? } } Thanks for your comments, } } Karen Pawlowski, Ph.D. } }
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
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Gray, Peter, 1975, Microtomist's Formulary and Guide, R. Krieger, Huntington, NY. (Still available for the hardy bibliophile at http://WWW.WEB4U.COM/cgi-bin/full_page?0-88275-247-2) reports FOUR fixative mixtures that are attributed directly to Champy:
My favorite, of course, is number 2!!! whose last component provides a wonderful bit of chemical history. We see how Bayer must have started. Dupont.
Regards, and I know that this doesn't help at all!
But it is all part of the art.
Fred Monson
} ---------- } From: Richard Edelmann } Reply To: edelmare-at-muohio.edu } Sent: Wednesday, September 5, 2001 4:24 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Bio. Fixation: What is Champy's Fluid? } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } } "Fixed 2-5 minutes in Champy's fluid, rinsed 2x in water" . . . followed } by } ETOH dehydration, CPD, mounting and SEM viewing. } } } O.k., we give up, searched numberous books, papers, catalogs, web } sites, etc. . . . does anyone have a recipe for "Chmapy's fluid" a.k.a. } "Champy's Solution"? Please? } } Thank you much! } } Richard E. Edelmann, Ph.D. } Electron Microscopy Facility Supervisor } 352 Pearson Hall } Miami University, Oxford, OH 45056 } Ph: 513.529.5712 Fax: 513.529.4243 } E-mail: edelmare-at-muohio.edu } } "RAM disk is NOT an installation procedure." } }
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I am investigating EDS systems to integrate with a JEOL-6100 SEM. I'm leaning towards PGT's Spirit system. Other considerations are Oxford's INCA and Thermo Noran's Vantage. Does anyone have particularly positive or negative experience with PGT? I don't often hear much about that company. Any comments about the other possibilities would be appreciated, as well.
Thanks!
-Peggy
Peggy McKarns Macatangay, PhD Project Analyst 2 Celanese Corpus Christi Technical Center
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (john_bruss-at-bose.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, August 22, 2001 at 17:00:52 ---------------------------------------------------------------------------
Email: john_bruss-at-bose.com Name: John Bruss
Organization: Bose Corp.
Education: Graduate College
Location: San Diego, CA, USA
Question: What materials and process would an amateur use to suspend a spider in a clear cube for unmagnified artistic and historic use? Where are those materials available in small quantities?
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I am pretty sure that Alwyn is not talking about x-ray mapping. If that was all he wanted, I suppose the key disk is all that would be needed if he already has the other imaging applications and hardware installed.
Someone suggested using Auto to scan an image for multiple spectra. Indeed, the software and hardware should allow it. The spectra might then be exported and processed by some other package to perform the type of component analysis that has been described at MSA over the last few years. My current ISIS x-ray application (version 3.32) has the option of storing the x-ray data in single column MSA format. Previous versions placed multiple channels on a single line.
One hitch would be the limitation on the number of spectra per job. I think there is a limit of 1000 or so built into the database used for managing the data. I suppose it could be changed to another number, but we bumped into it some time back.
That would mean you could do a 32x32 raster of points saving a spectrum at each. A four second acquisition per point would mean a bit more than an hour for acquisition. That would not be too bad, but the spatial resolution would probably be too low to be very useful. But the data should readily fit onto hard drives with capacities of a few gigabytes.
I would be interested in seeing how the data processing is handled. If someone makes it work, I would be very interested in hearing the details.
Warren
At 10:33 AM 8/23/2001 -0400, you wrote: } -----------------------------------------------------------------------. } This is correct, the keydisk is all that is required for } the software to be enabled. However, there may be hardware } required, too, at least a cable interface (RS232?) to the } microscope. I believe the ISIS Autobeam likes to select } its own scan rates. You may need to speak with Oxford re: } compatibility with your particular microscope. } } Matt } } Matthew J. Lynn } Center for Advanced Microscopy } University of Miami } (305)284-4736 } mlynn-at-miami.edu } } } On Wednesday, August 22, 2001 9:23 PM, Fred Pearson } [SMTP:eoptics-at-mcmail.cis.mcmaster.ca] wrote: } } } } Alwyn: } } } } By spectral images, do you mean xray mapping? A "key" disk for the ISIS } } software program is required to activate the xray mapping facility. Of, } } course this will cost a bit of money to purchase from Oxford. When the } } ISIS program is loaded initially, all the software required to use the } } program is there, but activating the unpaid for parts of the program is } } the thing. } } } } Fred } } } } On Wed, 22 Aug 2001, Alwyn Eades wrote: } } } We have an Oxford ISIS EDS system. As it stands it is not set up to } } } acquire spectral images. We would like to be able to do spectral image } } } acquisition on this machine. Is there anyone out there who knows how } } } the system may be modified (macros, interfaces, whatever) to do it? We } } } do have Digital Micrograph (on a Mac whereas ISIS is on a PC), if that } } } helps. } } } .......... } } } Alwyn Eades } } } Department of Materials Science and Engineering } } } Lehigh University } } } 5 East Packer Avenue } } } Bethlehem } } } Pennsylvania 18015-3195 } } } Phone 610 758 4231 } } } Fax 610 758 4244 } } } jae5-at-lehigh.edu
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Mike, My guess is that either will do the trick, but I would take EDX as a shortened version of Energy Dispersive Analysis of X-rays, which, of course, is EDAX. I've also seen EDXA which would keep you out of trouble with EDAX. One really is doing a specific type of analysis, spectroscopy, which would cause me to lean towards Energy Dispersive Spectroscopy.
Ken Converse owner Quality Images third party SEM service Delta, PA
Ingram, Mike wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Not a serious question here, more of a curiosity. } } I use EDS when talking about x-ray analysis. I see many using EDX. Which } is correct? } } Mike Ingram } } }
Dear Listers, In a clear out of a cupboard I have found a pair of Debye-Sherrer cameras and accessories. If anyone would like them, please contact me - otherwise they're skip fodder. I used to demo this as an undergraduate lab some years ago, but I guess no-one uses them any more?
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The CCD camera (model 679) in our CM20 TEM does not work any more. It is the fist commercial CCD camera of Gatan. The company is experiencing hard time to find the right replacing part. A programmable chip ( TEK1B2FV9 made by ACTEL) for the camera controller is needed. Then they can possibly figure out what problems with the camera.
If you have this part or any information which can help us to sole this problem please let me know. I can be reached at (613) 990-1403 or dashan.wang-at-nrc.ca
Try Prudence Rice's book Pottery Analysis: A Sourcebook (1987). This was the text on archaeological ceramics a few years back, and would be a good starting point. It is available for $65 on amazon.com.
Dear Katelyn, I will only add to Nelson Conti's remarks that you should be able to buy "blood-agar plates," which are Petri dishes already filled with a sterile, general bacterial culture medium, as well as sterile cotton-tipped swabs, from a medical supply store in your area. My daughter did a project along the same lines as yours and that's where we got the supplies. The dishes came in packs of 10, I think and they only cost a few dollars. We kept the inoculated dishes in a warm place. Around body temperature (37 degrees C, 98-99 degrees F) is ideal is you can find such a place. If you find bacterial colonies that have a clear ring around them, instead of the red of the rest of the blood agar, that is an indication that the bacteria in that colony are pathogenic (disease-producing). Good luck, and have fun.
Jill Verlander Reed, D.V.M. Associate Scientist Director, College of Medicine Electron Microscopy Core Facility University of Florida P.O. Box 100215 Gainesville, FL 32610 verlaj-at-medicine.ufl.edu Phone: (352) 846-0820 Fax: (352) 846-3299
At UIC we are less than 800m from the Blue Line of the cta L. When the lab was set up we bought an EMF cancellation system, in case. However, it turned out we did not need it despite doing high resolution annular dark field STEM with sub 0.2nm resolution.
While I was working for VG we did install a HB501 STEM in Halle, Germany. Here it was a different story. The main streetcar line through the city ran less than 100m from the site and to make matters worse a main feed for the system ran perpendicular to the line along a second wall of the building (probably 50m away). You could see each and every streetcar entering the stretch of line powered by the feed, it caused a sideways image jump! This was fixed by putting a Hall effect coil in the room at microscope bench level with a feed into the alignment coils.
If this is a new transit system then you are likely to suffer less problems with vibration and magnetic fields than you are with an old system where maintenance and standards of construction are likely to be poorer.
Alan
At 12:05 PM 9/7/2001 -0700, John C. Wheatley wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Alan W Nicholls, PhD Director of Research Service Facility (Electron Microscopy) Research Resources Center - East (M/C 337) Room 100 Science and Engineering South Building The University of Illinois at Chicago 845 West Taylor St Chicago, IL 60607-7058
Make sure that you withdraw the specimen from the polishing solution as quickly as possible (!) and rinse immediately (!). I don't know what you are using to rinse your specimens. If your electropolishing solution is based on methanol, than it is a good idea to rinse it in methanol. I normally rinse my specimens twice in two different beakers with methanol or ethanol. Good luck!
"yan.hu" {yan.hu-at-brunel. To: microscopy-at-sparc5.microscopy.com ac.uk} cc: Subject: ask help
09/11/01 03:35 AM
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Dear Listers,
I got a problem when preparing TEM foils of Al-alloy by means of electro-polishing. A dark film produced on the specimen surface when it was been polishing. I think, the polishing condition is right and listed below:
Solution, 25 % Nitric acid plus 75 % Mechenol Temperature, about -30 degree C Voltage, 15-20V
I would be pleased to hear from anyone who has an idea to sort out this problem.
This is the second round. I have not received enough responses to give any meaningful data. If you are interested in the results of this survey, but have not responded for your facility, please do so. Thanks!
This is a survey that was passed out at the Facility Managers Meeting at this year's M&M meeting. We've only gotten 6 responses so far, and many more are needed for valid results (although obviously only one answer per facility). Please take a couple of minutes to fill this form in, and email back to me. Thank you!
Phil
MICROSCOPY FACILITY SUPPORT
This survey will help compile the nature and amount of support provided to microscopy facilities by universities, colleges, and research institutes. We would like to determine how much facility support is provided by the institution, and how much must be generated by user fees or other sources, on a per cent basis.
If we get enough responses to have valid data, we will post the results on the microscopy listserver and send them to the MSA bulletin. Names of institutions will not be included in the reported results.
Institution: Location: Nature of institution (Academic, Commercial, private research, etc.)
Type of Facility (institution core, satellite, departmental, etc):
(Place an 'X' by the appropriate answer) Predominately: Biological Materials Both
User base: Service Multi-user Both
Type of microscopes: SEM FESEM ESEM or LV TEM FETEM Major optical (Specify) Scanning Probe (specify) Other (specify)
Approximately what *per cent* of the funds for each of the following is provided by A) your institution, B) user fees, or C) grants
Salaries: A) B) C)
Instrument Maintenance (Service contracts, instrument insurance, etc): A) B) C)
Replacement of existing equipment: A) B) C)
Purchase of new equipment (not replacement for but addition to existing equipment): A) B) C)
Supplies: A) B) C)
Other operating expenses: A) B) C)
Are there other microscope facilities at your institution? if so, please indicate approximate number having electron microscopes. Ideally we would like a form filled out for each major facility (multiple instruments) at your institution if there are more than one.
Please include additional information such as novel funding ideas for your facility or for others at your institution.
Please email your responses to Philip Oshel peoshel-at-facstaff.wisc.edu -- }}}}}}}}}}}}}}}}}{{{{{{{{{{{{{{{{{ Philip Oshel Supervisor, AMFSC and BBPIC microscopy facilities Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
Many thanks to everyone who responded to my question about electropolishing. The details of my experiment are: 1. it is an Al-Mg-Si alloy. 2. it is a twin jet-polish by Tenupole 3. The flow rate I used was 6. 4. The black film possibly appeares at the beginning of the polishing not after the foil prepared. 5. I noted that the current is more than 500 mA this time, not about 100 mA as before. This is a difference. Is it due to the problem of the instruments.
Has anyone done serial sections of decalcified mouse ears? I have been given some that were decalcified in EDTA for about 10 days, post-osmicated, dehydrated, infiltrated and embedded in Spurr resin. A grad student did the orienation for embedding, but along what plane I do not know. Each orientation is a little different. Can anyone recommend a consistent method of embedding?
This message is not relevant to our normal topics of discussion but I hope that under the circumstances you, Nestor, and others will forgive me for this once-off breach of the rules.
I am sure I speak on behalf of all subscribers to the MSA listserver when I express heartfelt shock and indignation at yesterdays tragedies in New York City, Washington DC and near Pittsburg. Those of us from elsewhere in the world assure our colleagues in the US that we share your grief, and offer our support at this difficult time. We extend our sincere condolences to those who have lost family members, friends and colleagues and we wish a speedy recovery to those who have been injured.
God bless, vasbyt and sterkte*.
Rob
* loosely translated = "hang in there and be strong"
======================================================= Robin H Cross Director : Electron Microscopy Unit, and Chairman : 15th International Congress on Electron Microscopy (ICEM-15) Rhodes University, PO Box 94, Grahamstown, South Africa tel: +27 46 603 8168/9, fax: +27 46 622 4377 http://www.ru.ac.za/emu/index.htm http://www.icem15.com =========================================================================== Remember that ICEM-15 takes place in Durban, South Africa in September 2002 ===========================================================================
I have watched this conversation with interest and must admit that I am a very nervous about a student in a High School isolating pathogens on blood agar at 37 deg C. This pre-supposes that there are trained microbiologists, safe handling facilities and disposal methods for unknown human pathogens.
If someone was to perform a risk assessment on this activity would they even consider letting an unsupervised first year degree student isolate unknown pathogens in the microbiology department?
Malcolm Haswell e.m. unit School of Sciences University of Sunderland UK
Jill Verlander Reed wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Katelyn, } I will only add to Nelson Conti's remarks that you should be able to buy } "blood-agar plates," which are Petri dishes already filled with a } sterile, general bacterial culture medium, as well as sterile } cotton-tipped swabs, from a medical supply store in your area. My } daughter did a project along the same lines as yours and that's where we } got the supplies. The dishes came in packs of 10, I think and they only } cost a few dollars. } We kept the inoculated dishes in a warm place. Around body temperature } (37 degrees C, 98-99 degrees F) is ideal is you can find such a place. } If you find bacterial colonies that have a clear ring around them, } instead of the red of the rest of the blood agar, that is an indication } that the bacteria in that colony are pathogenic (disease-producing). } Good luck, and have fun. } } Jill Verlander Reed, D.V.M. } Associate Scientist } Director, College of Medicine Electron Microscopy Core Facility } University of Florida } P.O. Box 100215 } Gainesville, FL 32610 } verlaj-at-medicine.ufl.edu } Phone: (352) 846-0820 } Fax: (352) 846-3299
=============================================================== ORIGINAL MESSAGE:
Email: sbcook-at-bcpl.net Name: Katelyn Cook
Organization: St. Michael the Archangel
Education: 6-8th Grade Middle School
Location: Baltimore, MD
Question: I am planning to do a science fair project about germs and the importance of hand washing. I want to collect germs from common public places such as doorknobs, handrails, telephones, sink faucets, etc. What is the best way to collect the germs? Will I need to grow the germs in a petri dish after I collect them? Will I be able to identify different types of germs using my microscope?
Hi everyone! I wonder if this is the right place for my doubt... I need if someone could tell me whether a major and important difference exists between Paraformaldehyde 4% vs Formaldehyde4% as a fixative for brain tissue. Thank you :) Valeria Burgos
_________________________________________________________ ¿Lo probaste? Correo gratis y para toda la vida en http://correo.yahoo.com.ar
I embed and section mouse cochleas on a regular basis (anywhere between 1 and 4 microns in thickness). Is your tissue dissected down to the spiral-shaped bone of the cochlea itself? Do you want sections along the plane of the modiolus (the center columnar structure of the cochlea) so that you get a cross-sectional view of three to five organ of Corti profiles? The cochleas I embed still have some vestibular structures remaining.
If so, I embed the cochleas in flat silicone molds with the flat surface of bone (which faces the brain) facing the bottom of the mold and the spiral surface up. After the resin has been polymerized and the blocks removed from the mold, I remount the block onto flat-ended cylinder blanks. (These are made by closing the cap of a BEEM capsule, cutting off the pyramidal tip, placing them with the closed cap down, filling them to the rim with resin, polymerizing and removing the BEEM capsule.) The block is remounted (I usually use SuperGlue) onto the blank with the cochlea side down (the curved surface now faces down and the flat bony surface faces up). Excess resin is removed by trimming the block as usual. When I'm cutting into the block, I continually check the orientation. I know I'm close the proper orientation if I start to get profiles of the maculae and/or cristae in the vestibular structures and a base-to-apex view of the modiolus. Continue to adjust until you get the desired orientation.
I hope this gets you started.
Jaclynn M. Lett, Research Technician III jlett-at-cid.wustl.edu
Fay and Carl Simons Center for Biology of Hearing and Deafness Central Institute for the Deaf 4560 Clayton Avenue St. Louis, MO 63110
voice: 314-977-0257 fax: 314-977-0030
-----Original Message----- } From: stacey andringa [mailto:andrina-at-email.uc.edu] Sent: Tuesday, September 11, 2001 1:52 PM To: Microscopy-at-sparc5.microscopy.com
Has anyone done serial sections of decalcified mouse ears? I have been given some that were decalcified in EDTA for about 10 days, post-osmicated, dehydrated, infiltrated and embedded in Spurr resin. A grad student did the orienation for embedding, but along what plane I do not know. Each orientation is a little different. Can anyone recommend a consistent method of embedding?
We have been working with hemagglutination tests lately (something new for our EM lab), and are looking for a quick and easy way to score fields captured using a standard LM with a digital camera.
We have tried some of the image analysis packages available, but have not found them very easy to use. Also we have some questions about the images themselves - how many blood cells have to be stuck together before it's considered to be a clump? A clump is a 3 dimensional shape, so this has to be taken into account, doesn't it?
If anyone is doing this routinely and wouldn't mind giving us a few hints (or references), we'd really appreciate it.
Thanks in advance for any help you can give.
Paula.
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
The term of "Spectral Imaging" is not well defined yet. If it means the distribution of certain number of elements on the surface of sample, that is the X-ray mapping. If it means all of possibility on the element table (means energy spectrum) on the entire surface of sample (mapping), I do not think the Oxford ISIS can do it. The simple way is to use the "Area Analysis" in stead of point analysis. That will give you all element signature on the selected area.
In ISIS 300, there is an EBSD package attached (need to get key, again), this package can do the similar thing. It collects the electron backscatter diffraction pattern (EBSP) on a single point and compares it with a signed crystallographic pattern (one needs to be known and signed). It does this on an array point by point and displays the matrix as a map. The final result shows the contrast of crystallography on the surface layer ( {50nm). This can be called as the imaging of diffraction property of given crystal. It is a very good tool for the study of grain size and boundary.
It is up to user's interest, what is your goal for "spectral imaging". ISIS has another software package, call Particle Analyzer (Gun shot analyzer). Again, user needs to give information about the most possible present elements, and the software can do the rest to see if these elements are shown up. Without these pre-defined information, software will have no direction to go and have no goal to be achieved.
Regards,
Zhiyu Wang Maxtor Corp.
-----Original Message----- From: Zhiyuw [mailto:zhiyuw-at-home.com] Sent: Tuesday, September 11, 2001 9:51 PM To: zhiyu_wang-at-maxtor.com Subject: Fw: Spectral Imaging with ISIS
----- Original Message ----- From: "Warren E Straszheim" {wesaia-at-iastate.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, August 23, 2001 10:32 AM Subject: RE: Spectral Imaging with ISIS
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am pretty sure that Alwyn is not talking about x-ray mapping. If that was } all he wanted, I suppose the key disk is all that would be needed if he } already has the other imaging applications and hardware installed. } } Someone suggested using Auto to scan an image for multiple spectra. Indeed, } the software and hardware should allow it. The spectra might then be } exported and processed by some other package to perform the type of } component analysis that has been described at MSA over the last few years. } My current ISIS x-ray application (version 3.32) has the option of storing } the x-ray data in single column MSA format. Previous versions placed } multiple channels on a single line. } } One hitch would be the limitation on the number of spectra per job. I think } there is a limit of 1000 or so built into the database used for managing } the data. I suppose it could be changed to another number, but we bumped } into it some time back. } } That would mean you could do a 32x32 raster of points saving a spectrum at } each. A four second acquisition per point would mean a bit more than an } hour for acquisition. That would not be too bad, but the spatial resolution } would probably be too low to be very useful. But the data should readily } fit onto hard drives with capacities of a few gigabytes. } } I would be interested in seeing how the data processing is handled. If } someone makes it work, I would be very interested in hearing the details. } } Warren } } At 10:33 AM 8/23/2001 -0400, you wrote: } } -----------------------------------------------------------------------. } } This is correct, the keydisk is all that is required for } } the software to be enabled. However, there may be hardware } } required, too, at least a cable interface (RS232?) to the } } microscope. I believe the ISIS Autobeam likes to select } } its own scan rates. You may need to speak with Oxford re: } } compatibility with your particular microscope. } } } } Matt } } } } Matthew J. Lynn } } Center for Advanced Microscopy } } University of Miami } } (305)284-4736 } } mlynn-at-miami.edu } } } } } } On Wednesday, August 22, 2001 9:23 PM, Fred Pearson } } [SMTP:eoptics-at-mcmail.cis.mcmaster.ca] wrote: } } } } } } Alwyn: } } } } } } By spectral images, do you mean xray mapping? A "key" disk for the ISIS } } } software program is required to activate the xray mapping facility. Of, } } } course this will cost a bit of money to purchase from Oxford. When the } } } ISIS program is loaded initially, all the software required to use the } } } program is there, but activating the unpaid for parts of the program is } } } the thing. } } } } } } Fred } } } } } } On Wed, 22 Aug 2001, Alwyn Eades wrote: } } } } We have an Oxford ISIS EDS system. As it stands it is not set up to } } } } acquire spectral images. We would like to be able to do spectral image } } } } acquisition on this machine. Is there anyone out there who knows how } } } } the system may be modified (macros, interfaces, whatever) to do it? We } } } } do have Digital Micrograph (on a Mac whereas ISIS is on a PC), if that } } } } helps. } } } } .......... } } } } Alwyn Eades } } } } Department of Materials Science and Engineering } } } } Lehigh University } } } } 5 East Packer Avenue } } } } Bethlehem } } } } Pennsylvania 18015-3195 } } } } Phone 610 758 4231 } } } } Fax 610 758 4244 } } } } jae5-at-lehigh.edu } } }
Wageningen University Dept Agrotechnology and Food Sciences
Postdoctoral Research Position in Cell Wall Microscopy
A vacancy for a Postdoctoral Position is available in the area of microscopy and functionality of cell walls as part of the research program of Dr. Henk Schols, Laboratory of Food Chemistry. The successful candidate will be studying serum separation in tomato products and the influence of processing at the cell wall level. Ideally immunolocalization techniques will be used to localise specific polysaccharides in these samples.
Candidates must originate from outside The Netherlands and hold a recent Ph.D. in a related field. The appointment is for 9 months and is available immediately.
For more information, interested candidates should contact:
Dr. Henk Schols Wageningen University Dept Agrotechnology and Food Sciences Laboratory of Food Chemistry Bomenweg 2 6703 HD Wageningen The Netherlands
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
Here is Washington, DC things are starting to get back to normal. Though what is normal has now changed. Our city was shut down for one day by cowardly terrorist acts. We are back at work and are carrying on. We will not let this break our spirit. The "Sleeping Giant" has been aroused and it is much larger than the perpetrators of this act realized as the Giant includes the global community.
Courage,
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
} } } "R. Cross" {r.cross-at-ru.ac.za} 09/12/01 03:19AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear friends and colleagues
This message is not relevant to our normal topics of discussion but I hope that under the circumstances you, Nestor, and others will forgive me for this once-off breach of the rules.
I am sure I speak on behalf of all subscribers to the MSA listserver when I express heartfelt shock and indignation at yesterdays tragedies in New York City, Washington DC and near Pittsburg. Those of us from elsewhere in the world assure our colleagues in the US that we share your grief, and offer our support at this difficult time. We extend our sincere condolences to those who have lost family members, friends and colleagues and we wish a speedy recovery to those who have been injured.
God bless, vasbyt and sterkte*.
Rob
* loosely translated = "hang in there and be strong"
======================================================= Robin H Cross Director : Electron Microscopy Unit, and Chairman : 15th International Congress on Electron Microscopy (ICEM-15) Rhodes University, PO Box 94, Grahamstown, South Africa tel: +27 46 603 8168/9, fax: +27 46 622 4377 http://www.ru.ac.za/emu/index.htm http://www.icem15.com =========================================================================== Remember that ICEM-15 takes place in Durban, South Africa in September 2002 ===========================================================================
I work on the University side of a large Medical center and yesterday after the attack in New York and Washington we received a broadcast e mail from the administration asking for blood. A couple of hours later I got to the blood bank and found a huge mob of medical workers in line to donate blood. Today the line was more then three hours long. I am going to wait a couple of days before donating blood. I think its realalistic that the demand for blood will be very high for at least a few weeks to come. I urge all healthy adults to join me in donating blood as its the only real thing that private citizens can do to help in this unprecedented catastrophe. Sincerely, Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Thomas Jefferson University Room 229 JAH 1020 Locust Street Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cell Timothy.Schneider-at-Mail.TJU.edu
There are a couple of ways to reduce the orientation variation. One way is to use flat-bottomed Beem capsules for embedding molds (I haven't got the order info on these with me, but I can get it to you later, or you may get a reply from someone who has the info handy). These molds have a small enough surface area at the bottom that you can place the bulla or inner ear in them, say medial side down and they don't rock or float out of position. (Fill the mold before manipulating the specimen.)
Another way I have for embedding the cochlea is to split it in half, down the modiolus just prior to embedding and place the cut surface face down in the molds. I use half of a double edged razor blade to split it under a dissecting scope and I orient the cut from the apex to the base with the round window membrane perpendicular to the top surface. If you process the cochlea to this point before you split it, it is much firmer and tougher to work with and you won't lose as much tissue, but you do lose some. Hope this helps.
Karen Pawlowski, Ph.D.
stacey andringa wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Has anyone done serial sections of decalcified mouse ears? I have been } given some that were decalcified in EDTA for about 10 days, } post-osmicated, dehydrated, infiltrated and embedded in Spurr resin. A } grad student did the orienation for embedding, but along what plane I do } not know. Each orientation is a little different. Can anyone recommend } a consistent method of embedding? } } Stacey.Andringa-at-uc.edu
I find your comment interesting as I have volunteered to judge several Regional and National Science Fairs for high school aged students and have seen this particular experiment carried out by a number of the students. The students that I choose to judge are actually the younger group, who are "middle school" aged and I doubt the majority of them are getting much outside help, other than their science teachers or they would have picked a topic that was more unique (or geared to what the expert was doing).
I'm sure the petri dishes never leave the science lab. and the students were gloves while handling the cultures, but that is probably the extent of it....
Karen Pawlowski, Ph.D.
Malcolm Haswell wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I have watched this conversation with interest and must admit that I am } a very nervous about a student in a High School isolating pathogens on } blood agar at 37 deg C. This pre-supposes that there are trained } microbiologists, safe handling facilities and disposal methods for } unknown human pathogens. } } If someone was to perform a risk assessment on this activity would they } even consider letting an unsupervised first year degree student isolate } unknown pathogens in the microbiology department? } } Malcolm Haswell } e.m. unit } School of Sciences } University of Sunderland } UK } } Jill Verlander Reed wrote: } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Dear Katelyn, } } I will only add to Nelson Conti's remarks that you should be able to buy } } "blood-agar plates," which are Petri dishes already filled with a } } sterile, general bacterial culture medium, as well as sterile } } cotton-tipped swabs, from a medical supply store in your area. My } } daughter did a project along the same lines as yours and that's where we } } got the supplies. The dishes came in packs of 10, I think and they only } } cost a few dollars. } } We kept the inoculated dishes in a warm place. Around body temperature } } (37 degrees C, 98-99 degrees F) is ideal is you can find such a place. } } If you find bacterial colonies that have a clear ring around them, } } instead of the red of the rest of the blood agar, that is an indication } } that the bacteria in that colony are pathogenic (disease-producing). } } Good luck, and have fun. } } } } Jill Verlander Reed, D.V.M. } } Associate Scientist } } Director, College of Medicine Electron Microscopy Core Facility } } University of Florida } } P.O. Box 100215 } } Gainesville, FL 32610 } } verlaj-at-medicine.ufl.edu } } Phone: (352) 846-0820 } } Fax: (352) 846-3299 } } =============================================================== } ORIGINAL MESSAGE: } } Email: sbcook-at-bcpl.net } Name: Katelyn Cook } } Organization: St. Michael the Archangel } } Education: 6-8th Grade Middle School } } Location: Baltimore, MD } } Question: I am planning to do a science fair project about germs and } the importance of hand washing. I want to collect germs from common } public places such as doorknobs, handrails, telephones, sink faucets, } etc. What is the best way to collect the germs? Will I need to grow } the germs in a petri dish after I collect them? Will I be able to } identify different types of germs using my microscope? } } ---------------------------------------------------------------------------
To all our Amercian fellow microscopists and friends, The Australian Society for Electron Microscopy Inc sends its deepest heartfelt condolences to you all and your loved ones at this time of such tragic and unbelievable loss. Our warmest thoughts and prayers are with you all. Gerry Nash President ASEM Inc -- Geraldine (Gerry) Nash Electron Microscopist
EM Unit Australian Antarctic Division, Channel Highway, Kingston, Tasmania 7050 Australia Ph: + 61 03 62 323 322 Fax: + 61 03 62 323 351
Visit our EM Unit web site: http://www.aad.gov.au/science/ResearchResources/em_unit/default.asp and Australian Antarctic Division web site: http://www.aad.gov.au/
To all our Amercian fellow microscopists and friends, The Australian Society for Electron Microscopy Inc sends its deepest heartfelt condolences to you all and your loved ones at this time of such tragic and unbelievable loss. Our warmest thoughts and prayers are with you all. Gerry Nash President ASEM Inc -- Geraldine (Gerry) Nash Electron Microscopist
EM Unit Australian Antarctic Division, Channel Highway, Kingston, Tasmania 7050 Australia Ph: + 61 03 62 323 322 Fax: + 61 03 62 323 351
Visit our EM Unit web site: http://www.aad.gov.au/science/ResearchResources/em_unit/default.asp and Australian Antarctic Division web site: http://www.aad.gov.au/
Director, Interdepartmental Laboratories American Museum of Natural History Central Park West at 79th Street New York, NY 10024 USA
Email: avklaus-at-amnh.org Tel: 212-769-5977
On Thu, 13 Sep 2001, Gerry Nash wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } To all our Amercian fellow microscopists and friends, } The Australian Society for Electron Microscopy Inc sends its deepest } heartfelt condolences to you all and your loved ones at this time of } such tragic and unbelievable loss. } Our warmest thoughts and prayers are with you all. } Gerry Nash } President ASEM Inc } -- } Geraldine (Gerry) Nash } Electron Microscopist } } EM Unit } Australian Antarctic Division, Channel Highway, Kingston, Tasmania 7050 } Australia Ph: + 61 03 62 323 322 Fax: + 61 03 62 323 351 } } Visit our EM Unit web site: } http://www.aad.gov.au/science/ResearchResources/em_unit/default.asp } and } Australian Antarctic Division web site: http://www.aad.gov.au/ } }
To all our American colleagues: On behalf of the Microscopical Society of Ireland, I would like to extend deepest sympathy and condolences to those who have been affected directly or indirectly by the atrocious terrorist attacks on the American Nation.
Sincerely, Alexander Black, President, Microscopical Society of Ireland
Department of Anatomy National University of Ireland, Galway.
There is a great difference. Paraformaldehyde is a POLYMER of formaldehyde(HCHO). Paraformaldehyde will yield HCHO if you heat the dry powder to around 90 degrees C (NOT RECOMMENDED!!!). I am sending a document under separate cover that will explain, but briefly, there are three words that are important here. Formalin (commercial, 40% HCHO solution in HOH, that contains from 5-15% methanol to prevent the HCHO from polymerizing), Paraformaldehyde (the polymer, a white powder), and formaldehyde solutions (generated from paraformaldehyde suspensions in water or buffer, which I designate as "pHCHO"). The attachment to the following email will explain in greater detail. Good question!!!
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II CASI Home Page: http://darwin.wcupa.edu/casi/ South Church Street West Chester, PA, 19383 Office: SS024 Phone: 610-738-0437 FAX: 610-436-3036 eMail: fmonson-at-wcupa.edu Please call before visiting
I am looking for antibodies against double-stranded DNA, and eventually antibodies against histones. The two companies that I contacted, Biogenex and Pierce, discontinued its productions. Does anybody know for a supplier?
Tea
*************************************** Tea Meulia Research Scientist and Director Molecular and Cellular Imaging Center Ohio State University/OARDC 1680 Madison Ave. Wooster OH 44691
Dear All, We have had a last minute change in the program for our program for Microscopy and Digital Imaging on Sept. 27, 2001
1:15 "Imaging Tissue Morphogenesis in Zebrafish Embryos." Mark Cooper, University of Washington
Regards, Glen -- Glen MacDonald Microscopy and Imaging Facility University of Washington Core for Communication Research Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 glenmac-at-u.washington.edu (206) 616-4156 (206) 616-1828 fax **************************************************************************
C:} The box said "Requires Windows95 or better". So I bought a Macintosh. **************************************************************************
In regard to what is "appropriate", I share my own view.
I agree that science and this list server under these circumstances should be divorced from any shade of political statement.
The science community (emphasis on the community) is not divorced from emotion, thank goodness.
I believe we graciously accept the simple gestures of sympathy and not imply insult or rejection.
Justification: It is a historical, if not still surreal and forever infamous, time for world civilization.
With best regards and appreciation, Ed
Angela Klaus {avklaus-at-amnh.org} on 09/13/2001 05:07:53 AM
To: Gerry Nash {gerry.nash-at-antdiv.gov.au} cc: microscopy-at-sparc5.microscopy.com
Thank you, Gerry.
Best regards,
Angela
Angela V. Klaus
Director, Interdepartmental Laboratories American Museum of Natural History Central Park West at 79th Street New York, NY 10024 USA
Email: avklaus-at-amnh.org Tel: 212-769-5977
On Thu, 13 Sep 2001, Gerry Nash wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } To all our Amercian fellow microscopists and friends, } The Australian Society for Electron Microscopy Inc sends its deepest } heartfelt condolences to you all and your loved ones at this time of } such tragic and unbelievable loss. } Our warmest thoughts and prayers are with you all. } Gerry Nash } President ASEM Inc } -- } Geraldine (Gerry) Nash } Electron Microscopist } } EM Unit } Australian Antarctic Division, Channel Highway, Kingston, Tasmania 7050 } Australia Ph: + 61 03 62 323 322 Fax: + 61 03 62 323 351 } } Visit our EM Unit web site: } http://www.aad.gov.au/science/ResearchResources/em_unit/default.asp } and } Australian Antarctic Division web site: http://www.aad.gov.au/ } }
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Listers, About a year ago there was a discussion about using ammonia to clean Wehnelts. I dutifully saved all the emails because the technique looked interesting. Of course, when I went looking for these emails I couldn't find them. I looked on the list archives but was not able to come up with anything. Could someone who uses this technique contact me off list?
Thanks -- Glenn
=============================================================================== Glenn Poirier 3450 University St, rm. 238 MicroAnalytical Laboratory Montreal, Qc Earth and Planetary Sciences tel (514) 398 6774 McGill University fax (514) 398 4680 email: glennp-at-eps.mcgill.ca http://castaing.eps.mcgill.ca
++ Millenium hand and shrimp ++ ===============================================================================
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Thank you, all, for your support. I would like to share an article I received with everyone, especially my fellow Americans. I hope nobody is offended by it, for I do not intend, or wish, for that to happen.
Darrell
} I know you are all receiving a lot of emails like this today...but read } on: } } Subject: This, from a Canadian newspaper, is worth sharing. } } America: The Good Neighbor. } } Widespread but only partial news coverage was given recently to } remarkable editorial broadcast from Toronto by Gordon Sinclair, } a Canadian television commentator. } What follows is the full text of his trenchant remarks as } printed in the Congressional Record: } } "This Canadian thinks it is time to speak up for the Americans as } the most generous and possibly the least appreciated people } on all the earth. } Germany, Japan and, to a lesser extent, Britain and Italy were } lifted out of the debris of war by the Americans who poured in } billions of dollars and forgave other billions in debts. } None of these countries is today paying even the interest } on its remaining debts to the United States. } } When France was in danger of collapsing in 1956, it was the } Americans who propped it up, and their reward was to be insulted } and swindled on the streets of Paris. I was there. I saw it. } } When earthquakes hit distant cities, it is the United States } that hurries in to help. This spring, 59 American communities } were flattened by tornadoes. Nobody helped. } } The Marshall Plan and the Truman Policy pumped billions of dollars } into discouraged countries. Now newspapers in those countries are } writing about the decadent, warmongering Americans. } } I'd like to see just one of those countries that is gloating over } the erosion of the United States dollar build its own airplane. } Does any other country in the world have a plane to equal } the Boeing Jumbo Jet, the Lockheed Tri-Star, or the Douglas DC10? } If so, why don't they fly them? } Why do all the International lines except Russia fly American } Planes? } } Why does no other land on earth even consider putting a man or } woman on the moon? } You talk about Japanese technology, and you get radios. } You talk about German technology, and you get automobiles. } } You talk about American technocracy, and you find men on the } moon-not once, but several times-and safely home again. } You talk about scandals, and the Americans put theirs right } in the store window for everybody to look at. } } Even their draft-dodgers are not pursued and hounded. They are } here on our streets, and most of them, unless they are breaking } Canadian laws, are getting American dollars from ma and pa at home } to spend here. } } When the railways of France, Germany and India were breaking down } through age, it was the Americans who rebuilt them. } When the Pennsylvania Railroad and the New York Central went broke, } nobody loaned them an old caboose. } Both are still broke. } } I can name you 5000 times when the Americans raced to the help of } other people in trouble. Can you name me even one time when } someone else raced to the Americans in trouble? } I don't think there was outside help even during the San Francisco } earthquake. } } Our neighbors have faced it alone, and I'm one Canadian who is } damned tired of hearing them get kicked around. } They will come out of this thing with their flag high. } And when they do, they are entitled to thumb their nose at } the lands that are gloating over their present troubles. I hope } Canada is not one of those." } } Stand proud, America! } } I would hope that each of you would send this to as many people } as you can and emphasize that they should send it to as many } of their friends until this letter is sent to every person on the web.
Various methods for cleaning Wehnelt cylinders, and other internal parts of vacuum systems, are discussed in some detail in Section 2.10.4c (see bottom of page 72) of the book Vacuum Methods in Electron Microscopy (for info about this book see: http://www.2spi.com/catalog/books/book48.html and http://pup.princeton.edu/titles/6484.html).
Disclaimer: I make about $1.50 from each copy of this book sold. -- Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan 3062 Dow Bldg.; 2300 Hayward St. Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
To all those people who have expressed their concerns,
I would like to thank all of those people who have sent their heartfelt messages and words of encouragement. Especially now in our time of need, it is uplifting to know all of our fellow microscopists from other nations are expressing their concern and support. During this time, when it is difficult to sort out our emotions, all of your words have helped to restore some of my faith in humanity.
Thanks you. Steve
_________________________________________________________________ Get your FREE download of MSN Explorer at http://explorer.msn.com/intl.asp
I must humbly apologize for the "article" I sent. A coworker has informed me that it is a hoax. My first instinct was to not send it. I made the mistake of trusting the source, and allowing the video images of crowds of people cheering the deaths of tens of thousands of Americans in the matter of a few hours, cloud my better judgment.
I was, and am, sincere in my thanks for the support expressed on the list, and from all other sources.
Darrell
(These are my opinions. You can blame no one else for them.)
Usually we do our own SEM work, but a special job requirement may arise which I cannot accommodate using our equipment. It may be desirable to tensile fracture a thin ceramic sheet (predominately zirconia) and examine the fracture surface without breaking vacuum. EDS would likely be required as well. Magnification requirements *should* range from about 20x to 5000x. It would not surprise me, however, if 30-50 Kx might provide useful information.
If you have the equipment and take "outside" work, please contact me by direct email or phone.
Direct line to my office and lab: 434.522.6111 ...If A/C 434 fails, try the old one: 804
Thanks,
Woody White McDermott Technology, Inc. Lynchburg Research Center Lynchburg, VA
McDermott site: http://www.mtiresearch.com/ Personal site: http://woody.white.home.att.net
I am not sure is it original recipe or it's my modification, but anyway, the following works for me. I am sonicate Wehnelt cup in 1:1 diluted ammonia solution (stock ammonia solution I believe is 30%) for 10-15 min, rinse with deionized water and dry at +60 in the oven.
Good luck. Sergey
At 10:11 AM 9/14/01 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Wehnelt caps are usually made from stainless steel (304).
For JEOL wehnelts, I have had customers use "ammonium hydroxide hydrogen peroxide 30% by volume". I am not a chemist so I can't give you the particulars but this works great as it attacks everything except the stainless steel. I usually let the cap soak in this solution overnight.
Regards,
Earl weltmer ----- Original Message ----- } From: "Glenn Poirier" {glennp-at-eps.mcgill.ca} To: "Microscopy Listserver" {microscopy-at-sparc5.microscopy.com} Sent: Friday, September 14, 2001 7:11 AM
Dear American Fellow Microscopists, After the first moments of shock and grief let us express our deepest sorrow and sympathy to the American microscopists in particular and to the American people in general on behalf of all Hungarian microscopists. We have so many friends and colleagues there... To overcome these hard days and weeks let us wish you all strength both physically and in your soul, work and fight together for a better world! Kristof Kovacs
----------------------------------------------------------- Dr. Kristof Kovacs President, Hungarian Society for Microscopy University of Veszprem Veszprem, P.O.Box 158 H-8201 HUNGARY Phone: +36-(88)-421-684, +36-(30)-930-3931 Fax: +36-(88)-423-091
Food Structure & Functionality Symposium 2002 May 5 - 8, 2002, Palais des Congrès de Montréal o Montréal, Québec, Canada.
An international symposium leading Food Structure & Functionality studies through the 21st century "webaddress at the AOCS site (http://www.aocs.org/member/division/fsff/index.htm)"
Being held in conjunction with the 93nd AOCS Annual Meeting & Expo (www.aocs.org)
The symposium has two themes: * new and novel approaches (including microscopy, rheology and spectroscopy) to the study of structure-function relationships in foods; * food system studies covering any part of the processing chain - from the raw material to the final product, and including trouble shooting.
Tentative Program Schedule Sunday, May 5th Short Course - Understanding structure-function relationships in food systems through specific localisation methods and microscopy. Contact: Marcel Paques (Paques-at-nizo.nl)
Monday, May 6th Morning
Opening of symposium Plenary Speaker Dairy Applications Session. Chairs: Mark Auty (mauty-at-moorepark.teagasc.ie) and Harjinder Singh (H.Singh-at-massey.ac.nz) Lunch break Afternoon Colloidal and Interfacial Sciences Session. Chairs: Marcel Paques (Paques-at-nizo.nl) and David Pechak (Dpechak-at-kraft.com)
Dedicated Poster Session Division Board Meeting
Tuesday, May 7th Morning Agricultural Applications of Microscopy and Imaging Session./ joint with Feed Microscopy Division. Topic/Tentative title (will probably be revised): New Microscopic Techniques for Identifying Food/Feed Constituents and Contaminants contacts: Mark Auty (mauty-at-moorepark.teagasc.ie) and Kim Koch
Lunch break: Division Luncheon and round table (expert) discussion. Topic TBA Afternoon Microbiology and Food Session. Chairs Judy Arnold (jarnold-at-saa.ars.usda.gov) and Ida Yates (iyates-at-ars.usda.gov)
Food Structure and Functionality Forum - Division Members Meeting
Wednesday, May 8th Morning Ingredients and Food Processing Session. Chairs: Diana Kittleson (dkittleson-at-pillsbury.com) and Bernhard Tauscher (bernhard.tauscher-at-bfe.uni-karlsruhe.de)
Lunch break Afternoon New Methods and Techniques for Food Structure and Functionality Analysis Session. Chairs: Kathy Groves Kgroves-at-LFRA.co.uk and Maud Langton maud.langton-at-sik.se
Closure of Symposium.
*-------------------------------------------------------------------------------------------------------------------- For further information, please go to the websites indicated, contact session chairs listed above, or myself.
Paula.
Paula Allan-Wojtas Research Scientist - Food Microstructure Agriculture and Agri-Food Canada Atlantic Food and Horticulture Research Centre Kentville, Nova Scotia Canada B4N 1J5
Well, you hit the World Trade Center, but you missed America. You hit the Pentagon, but you missed America. You used helpless American bodies, to take out other American bodies, but like a poor marksman, you STILL missed America.
Why? Because of something you guys will never understand. America isn't about a building or two, not about financial centers, not about military centers, America isn't about a place, America isn't even about a bunch of bodies. America is about an IDEA. An idea, that you can go someplace where you can earn as much as you can figure out how to, live for the most part like you envisioned living, and pursue Happiness. (No guarantees that you'll reach it, but you can sure try!)
Go ahead and whine your terrorist whine, and chant your terrorist litany: "If you cannot see my point, then feel my pain." This concept is alien to Americans. We live in a country where we don't have to see your point. But you're free to have one. We don't have to listen to your speech. But you're free to say one. Don't know where you got the strange idea that everyone has to agree with you. We don't agree with each other in this country, almost as a matter of pride.
We're a collection of guys that don't agree, called States. We united our individual states to protect ourselves from tyranny in the world. Another idea, we made up on the spot. You CAN make it up as you go, when it's your country. If you're free enough. Yeah, we're fat, sloppy, easygoing goofs most of the time. That's an unfortunate image to project to the world, but it comes of feeling free and easy about the world you live in. It's unfortunate too, because people start to forget that when you attack Americans, they tend to fight like a cornered badger. The first we knew of the War of 1812 was when England burned Washington, DC, to the ground. Didn't turn out like England thought it was going to, and it's not going to turn out like you think, either.
Sorry, but you're not the first bully on our shores, just the most recent. No Marquis of Queensbury rules for Americans, either. We were the FIRST and, so far, only country in the world to use nuclear weapons in anger. Horrific idea, nowadays? News for you bucko, it was back then, too, but we used it anyway. Only had two of them in the whole world and we used 'em both.
Grandpa Jones worked on the Manhattan Project. Told me once that right up until they threw the switch, the physicists were still arguing over whether the Uranium alone would fission, or whether it would start a fissioning chain reaction that would eat everything. But they threw the switch anyway, because we had a War to win. Does that tell you something about American Resolve? } So who just declared War on us? It would be nice to point to some real estate, like the good old days. Unfortunately, we're probably at war with random camps, in far-flung places. Who think they're safe. Just like the Barbary Pirates did. Better start sleeping with one eye open. There's a spirit that tends to take over people who come to this country, looking for opportunity, looking for liberty, looking for freedom. Even if they misuse it. The Marielistas that Castro emptied out of his prisons, were overjoyed to find out how much freedom there was. First thing they did when they hit our shores, was run out and buy guns. The ones that didn't end up dead, ended up in prisons. It was a big PITA then (especially in south Florida), but you're only the newest PITA, not the first.
You guys seem to be incapable of understanding that we don't live in America, America lives in US! American Spirit is what it's called. And killing a few thousand of us, or a few million of us, won't change it. Most of the time, it's a pretty happy-go-lucky kind of Spirit. Until we're crossed in a cowardly manner, then it becomes an entirely different kind of Spirit.
Wait until you see what we do with that Spirit, this time. Sleep tight, if you can. We're coming. And when we arrive, well, let's just say it won't be pretty.
I have been given a piece of titanium that contains small precipitates of unknown composition and asked to prepare a TEM sample so that the precipitates can be analysed. As it is only the precipitates that are of interest I thought that the best approach would be to make an extraction replica. However, to do that need to find a way to etch away the titanium matrix while leaving behind the unknown precipitates.
I would be grateful for any recommendations for a suitable etchant that will preferentially remove the titanium matrix so that the extraction replica can be made.
Regards,
Martin Saunders. --
*****************************************
Dr. Martin Saunders, Lecturer, Centre for Microscopy and Microanalysis, University of Western Australia, Crawley, Western Australia 6009, Australia.
It has been very positive to see, on the microscopy listserver, messages from microscopy communities in other nations addressed to those of us who live and work in the United States offering sympathy for the tragic events of last Tuesday. These messages have, without exception, been measured, responsible and generous. We are grateful for them.
Speaking for myself, I want to say how offended I was to read, on the microscopy listserver, "Letter to a Terrorist" from Peter Jordan. There are no terrorists on this list server. This is not an appropriate place for this kind of polemic. I hope that those who take exception to Mr. Jordan's remarks will refrain from responding to the substance of what he says. The last thing we want is for the listserver to be taken over by a political debate on the issues raised by the attacks on New York and Washington.
Alwyn -- .......... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
I like to dissolve fat in insects bodies what could be the best chemical . benzine, carbon tetrachloride, or???
Keep care and be of good cheer.
Regards
(name) Vratislav Richard Eugene Maria John Baptist (surname) of Bejsak (Bayshark)-Colloredo-Mansfeld
website: http://www.coleoptera.org listserver: coleoptera on www.egroup.com/group/coleoptera/info.html Coleoptera - Australia, Tenebrionidae of World (incl. Lagriinae, Alleculinae)
University of Sydney The Wentworth Bldg., Box 62 NSW 2006 AUSTRALIA phone : +61 414 540 465 email: vratislav-at-bigfoot.com ricardo-at-ans.com.au (before Ricardo-at-compuserve.com and ricardo-at-login.cz )
Only after the last tree has been cut down, only after the last river has been poisoned, only after the last fish has been caught, only then will you find that money can not be eaten.' CREE INDIAN PROPHECY.
Incoming mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com).
Could anybody recommend the good method of sample preparation for SEM imaging on carbon nano tubes? The sample is a powder type including single wall carbon nanotubes (SWCNTs). Thank you very much in advance.
""} Subject: This, from a Canadian newspaper, is worth sharing. } } America: The Good Neighbor. } } Widespread but only partial news coverage was given recently to } remarkable editorial broadcast from Toronto by Gordon Sinclair, } a Canadian television commentator. } What follows is the full text of his trenchant remarks as } printed in the Congressional Record:""
Gordon Sinclair's piece was first broadcast in 1973 - see:
I'd like to thanks to all who has collaborate for the answers on tempers studies on potteries by SEM. I hope in brief get in touch with some of them.
I was trying to don't say a word on the terrorism subject, that because sometimes for me when something so horrible happens with me I prefer stay quiet on my side in order to recover my balance to overcome such pain. However, I would like to expreess my deep feelings to American people and gouvernement. Since the attacks I could not stop to think how terrible the human been can be, even if sometimes the streets can show us terrible facts. I've watched several chanels, brazilian and foreigners, and still can understand what hapens and why that people has died. I would like to remark my sympathy to your gouvernement which, in my oppinion, is doing their job in this difficult situation as well as possible. We hope you can find find a pacific way to solve this problem, however, if you can't don't mistake several other countries around the world, including Brazil, and I belive some arabian countries too, is supporting you and almost ready to stay beside you. You know the American spirit is not living only in US as well stand by Peter Jordan, this is the spirit that you can see in many other countries who respect the Democracy, The Human Rights, The Freedom, and have been proclamed in the France Revolution, US Constitution as well as in many other constitutions and UN Declarations. Again, here in Brazil we're deeply touched by your pain and we hope help you as better as possible to overcome this terrible incident.
The Ohio State University Microscopic and chemical Analysis Research Center (MARC) anticipates an opening for an electron microscopist beginning October 1 to operate a scanning electron microscope (SEM) and an electron microprobe (EPMA). The position includes working with researchers, teaching students the basic theory and practical SEM and EPMA measurement techniques, collaborating on research projects and directing staff assistants. We seek an excellent microscopist who enjoys the exciting atmosphere of a major research university and will enjoy collaborating with scientists on a broad variety of projects. The MARC serves the entire Ohio State University as well as other institutions, with projects from diverse research areas including geology, materials science, environmental sciences, biological and biomedical sciences, dentistry, textiles. The MARC focuses on elemental analysis using from microscale to bulk analysis using SEM, EPMA, inductively coupled plasma optical emission spectrometry and inductively coupled mass spectrometry with laser ablation or solution sampling. The MARC also teaches traditional or short courses on each of the techniques. The position is a full time, hard money funded job.
If you know of a suitable candidate for this position, please contact Dr. John Olesik, MARC Director, at olesik.2-at-osu.edu {mailto:olesik.2-at-osu.edu} or (614) 292-6954. Excellent candidates with a range of experience and potential to grow will be considered.
---------------------------------------------------------------------------- -- John Olesik Adj. Assoc. Professor, Research Scientist MARC Director Microscopic and chemical Analysis Research Center Ohio State University 125 S. Oval Mall 275 Mendenhall Laboratory Columbus, OH 43210
The "article" you forwarded is a hoax in that it was originally broadcast in 1973 and the author Gordon Sinclair died in 1984. However, many will agree that the ideas in the broadcast apply as well today as they did then.
We have a used Amray 1000A with a PGT IMIX EDS that we will part with, and are seeking best offer. The equipment details are on the used equipment page of the MSA web site.
I am currently recruiting for a Bristol (South West England) based high technology company which is seeking an experienced microscopist to join its pioneering team as a Lab Manager. I would like to know whether or not you might know of anybody suitably qualified who may be interested. My client will be happy to arrange Working Visa details. Regards.
Hello Haeseong, If you are just looking at the nanotubes, it would be easy to place some of the powder on double-sided carbon tape or double sided Cu tape, and just put the sample in the SEM. The important thing is to have the machine very well aligned, and astigmatism corrected for a small spot size and a voltage of about 5kV or less. You should start seeing useable information at a magnifcation range of 30-50kx and above.
-Brad
---------- From: Haeseong Lee Sent: Monday, September 17, 2001 5:55 AM To: Microscopy-at-sparc5.microscopy.com Subject: SEM imaging on CNT
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
?? Hello.
Could anybody recommend the good method of sample preparation for SEM imaging on carbon nano tubes? The sample is a powder type including single wall carbon nanotubes (SWCNTs). Thank you very much in advance.
I seriously doubt that Mr. Jordan meant or implied that there were terrorists in the Listserver.
Much like the lamentations I hear on the radio seemingly directed at terrorists about how the terrorists have "awakened a sleeping giant" etc. I doubt the author meant that everyone listening to the radio is a terrorist.
I am sure it was meant as a public message to vent all of our outrage at this terrible event.
Regards,
Earl Weltmer
----- Original Message ----- } From: "Alwyn Eades" {jae5-at-lehigh.edu} To: "EMNET" {microscopy-at-sparc5.microscopy.com} Sent: Monday, September 17, 2001 5:35 AM
I'm not familiar with recent reviews or specific insect protocols, but there's an older book by Morris Kates called techniques in lipidology -part of a series of lab -type manuals. One of the more universal ways to dissolve / extract lipids is the Bligh-Dyer method (Can J Biochem & Physiol 1959 37:911-917 , paraphrased from Kates:
Extract the specimen with one volume of chloroform:methanol:water 1:2:0.8, which efficiently extracts lipids. the extract is then diluted with one volume each of chloroform and water tp form the two-phased system, chloroform and Methanol/water (1.0:0.9). Any water-soluble contaminants are thus partitioned into the methanol-water phase.
Carbon tet is probably NOT the best to extract lipids with -they are more soluble in chloroform
Richard
} "Vr. Richard Bejsak-Colloredo-Mansfeld" {ricardo-at-ans.com.au} 9/17/01 5:37:30 AM } } } Dear Colleagues
I like to dissolve fat in insects bodies what could be the best chemical . benzine, carbon tetrachloride, or???
Keep care and be of good cheer.
Regards
(name) Vratislav Richard Eugene Maria John Baptist (surname) of Bejsak (Bayshark)-Colloredo-Mansfeld
Hello folks. I am trying to fix and stain with lanthanum nitrate and have followed procedures in Hyatt's book. It mentions that lanthanum will precipitate in the cold, and when mixed with phosphate buffer. I have used cacodylate buffer, room temperature solutions and checked the pH of my buffer(7.2), but I am seeing a lot of ppt in all the solutions (glut fix, wash and osmium fix). I am wondering if anyone has any suggestions about how to alleviate this problem. Is the precipitate harmful to the tissue? Can I filter the solutions to get rid of big clumps? Is this normal? I would appreciate any feedback from someone who has used the stuff. Thanks, JoAnn
JoAnn Buchanan Molecular and Cellular Physiology Stanford University School of Medicine Stanford, CA 94305 650-723-5856
Anyone taking down a darkroom or willing to donate a good photographic processor for black and white prints to a Children's Hospital?
-- Jacob Bastacky, M.D. Research Physician Children's Hospital Oakland Research Institute 5700 Martin Luther King Jr. Way Oakland, California 94609 Telephone: 510.450.7639 email: jbastacky-at-CHORI.org FAX: 510.450.7910
Senior Scientist, Visiting National Center for Electron Microscopy Lawrence Berkeley National Laboratory University of California Berkeley, California 94720 sjbastacky-at-lbl.gov
I would like to hear some opinions about the Idefix EDS system, commercialized by SamX in France. Are they (probably french) List Members who know about it ? Please contact me off-list by mail.
Thanks
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess 67037 Strasbourg CEDEX France
Hope you are not knocked off from your normal life by the tragic events.
Anyway, I have a question for you gurus: we have a 2010F TEM sitting in a regular sized room. Somehow we feel there is a bit above normal acoustic echos going on in the lab which interferes with the TEM. So we hope to find some kind of insulation to be put on the wall and/or around the TEM to improve the performance.
Any thoughts and suggestions are highly appreciated.
Thanks and take care.
********************************* Chaoying Ni Electron Microscopy Facility College of Engineering University of Delaware Newark, DE 19716 *********************************
Dear Cathy, When I was at the Australian Electron Microscopy Meeting there were a lot of papers about wool, including extensive TEM. Perhapps Mel Gibson or someone at CSIRO (www.csiro.au) could help. At 09:55 AM 9/17/01 +1000, you wrote: } Hi, } } Can anyone recommend a method for preparing wool from sheep for TEM? Is it } easy to section? } } Thanks } } Cathy Gillespie } Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Dear Martin, Although I have not tried an extraction replica on Ti alloys myself, my Vander Voort lists several Ti alloy etchants, most based on combinations of HF asnd other strong mineral acids. The first is: "20 ml HCl 40 ml HF 40 ml H2O Immerse specimen in solution at 120 - 150 degrees F for 20-30 min. If smut forms immerse in 30% H2SO4 for 3 min." The simplest is: "50 ml HCl 50 ml H2O General-purpose macroetch (Ogden anbd Holden)." Hope this helps. At 06:40 PM 9/17/01 +0800, you wrote: } Dear all, } } I have been given a piece of titanium that contains small } precipitates of unknown composition and asked to prepare a TEM sample } so that the precipitates can be analysed. As it is only the } precipitates that are of interest I thought that the best approach } would be to make an extraction replica. However, to do that need to } find a way to etch away the titanium matrix while leaving behind the } unknown precipitates. } } I would be grateful for any recommendations for a suitable etchant } that will preferentially remove the titanium matrix so that the } extraction replica can be made. } } Regards, } } Martin Saunders. } -- Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchg.ubc.ca
Anyway, I have a question for you gurus: we have a 2010F TEM sitting in a regular sized room. Somehow we feel there is a bit above normal acoustic echos going on in the lab which interferes with the TEM. So we hope to find some kind of insulation to be put on the wall and/or around the TEM to improve the performance.
Any thoughts and suggestions are highly appreciated.
Dear Chaoying, One of the other listers will surely know a company which produces sound-dampening material, but while you wait for it to be delivered, try using either styrofoam or bubble wrap as a temporary solution. The styrofoam peanuts would probably work best, but they will take a lot of labor to install--although you might be able to put up something in front of the walls and backfill with them. Cloth drapes also work to dampen sound. Furthermore, you only need to be concerned with the frequency range which matches resonances in your scope, so when you purchase the insulation, be sure it will absorb those frequencies. Good luck. Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
Your comments about the European aviation industry are certainly true and I would expect that everyone on the list knows about the widespread use of the Airbus. Consequently, I did not think it was necessary to explain that I did not mean that the parts of the radio broadcast which include specific references to the "American technocracy" which are so clearly outdated should be considered as applicable today.
I completely agree with you that care must be taken regarding references to the Sinclair broadcast. In fact, in my original posting, I said "many will agree", which implies that "not all will agree". Also, I referred to the "ideas" of the broadcast, not the "specifics" of the broadcast. But as your e-mail points out, the ambiguity of the "ideas" can still lead to confusion, since they are left to personal interpretation.
Actually, I hesitated to respond to the original posting at all since it was clearly off-topic, however, the magnitude of the attack and the historical foundations of the "hoax", made me think that a short clarification regarding the original radio broadcast may be of general interest. I certainly did not mean to imply anything negative regarding the European (or any other country's) patriotism or spirit and I will be more careful regarding postings in the future that may have any nationalistic interpretations.
Joe _________________________________________ Joe Nabity, Ph.D. JC Nabity Lithography Systems E-Beam Lithography using Commercial SEMs & STEMs PO Box 5354, Bozeman, MT 59717 USA Voice: (406) 587-0848 FAX: (406) 586-9514 E-mail: info-at-jcnabity.com Web: www.jcnabity.com
For more information or to register, contact Lou Ross at rosslm-at-missouri or (573) 882-4777.
Central States Microscopy and Microanalysis Society Fall Meeting Friday, October 19, 2001 Adams Conference Center Veterinary Medicine Building University of Missouri Columbia, Mo 65211
8 - 8:45 Registration
8:50 Welcome
9:00 Tobias Baskin, University of Missouri-Columbia, "Ultrastructure of the Plant Cell Wall Analyzed with Field-Emission Scanning Electron Microscopy"
9:15 Nadia Navarrete-Tindall, University of Missouri-Columbia, "Morphology of the Endangered Legume Amorpha nitens Boynton and its Rhizobia Symbiont"
9:30 Heide Schatten, University of Missouri-Columbia, "Cytoskeletal Dynamics in Apicomplexan Parasites"
9:45 Joe Simmons, University of Missouri-Columbia, "Idiopathic Interstitial Pneumonia in Laboratory Rats"
10:00 Marty Katz, University of Missouri-Columbia, "Stem Cell Transplantation Therapy for Inherited Neurodegenerative Disorders"
10:15 Tom Phillips, University of Missouri-Columbia, "Fixation of Biological Tissues - Science or Art?"
10:30 - 10:45 BREAK
10:45 Robert Hall, Associate Vice Provost of Research, University of Missouri-Columbia, "The University of Missouri: Taking the Lead in National Research Funding"
11:00 Keynote speaker: Kenneth Moore, University of Iowa, MSA Tour Speaker, "Applications of Microscopy Techniques to the Study of Genetic Therapy Research"
12:00 - 1:30 Lunch
1:30 John Bozzola, SIU-Carbondale, "Multidisciplinary Approach to Teaching Electrion Microscopy"
1:45 Howard Wilson, University of Missouri-Columbia, "Creating a Scientific Poster Presentation using Microsoft Powerpoint"
2:00 Scott Walck, PPG, "Low Angle Cleavage - A New Specimen Preparation Method for TEM"
2:15 Louis Ross, University of Missouri-Columbia, "Exciting Times in Scanning Electron Microscopy and X-ray Microanalysis"
2:30 Vladimir Dusevich, University of Missouri-KC, "Practical ESEM"
2:45 Jeff Speakman, University of Missouri-Columbia, "Laser Ablation ICP-MS as a Tool for Characterization of Archaeological Materials"
3:00 - 3:15 BREAK
3:15 Cammy Bright, University of Missouri-Columbia, "A Laser Ablation ICP-MS and SEM Study of Rare Earth and Trace Elements in Conodonts"
3:30 Mike Amspoker, Westminster College, "A Light and Scanning Electron Microscopy Study of the Freshwater Diatom Desmogonium rabenhorstianum Grunow"
3:45 Heather Ramsay, University of Missouri-Columbia, "New Applications of SEM in Osteological Morphology Research"
4:00 Dickerson Moreno, University of Missouri, "Development of N-Type Diamond Semiconductor Through Field Enhanced Diffusion by Optical Activation"
4:15 Alejandro Suarez, University of Missouri, "A New Method to Fabricate Polycrystalline Diamond as a P-Type Semiconductor"
4:30 Closing Remarks
4:45 CSMMS Business Meeting
Senior Electron Microscope Specialist Electron Microscopy Core Facility W136 Veterinary Medicine University of Missouri Columbia, MO 65211 (573) 882-4777, fax 884=5414 email: rosslm-at-missouri.edu web: www.missouri.edu/~geosclmr/ebaf.html
I've just had the misfortune to smash the glass bell jar on my Edwards 306 coater. About 305mm dia, 400mm high.
While I can and maybe should buy a glass replacement, I'm tempted to make one from a 400mm length of 305mm diameter stainless-steel pipe with a flat top of 10mm thick polycarbonate.
I've never liked having my face too close to the glass one when it's in use, anyway.
Anyone got any comments/recommendations?
Anyone ever had a glass one implode in use?
cheers
rtch
Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
I have not known a glass tube to implode but we use an outer cover just in case (perspex tube). Tubes are checked for cracks before use and previous problems have been obvious, usually by the many bits all over the floor!
My worry would be the 10mm polycarbonate top, at 300mm dia that's over 1500 pounds (750Kg?) force. I would want something thicker or stronger.
I also like being able to look in the side to see my coating set up after evacuation and the evaporation (through appropriate goggles).
Regards, Ron
} I've just had the misfortune to smash the glass bell jar on my } Edwards 306 coater. About 305mm dia, 400mm high. } } While I can and maybe should buy a glass replacement, I'm tempted to } make one from a 400mm length of 305mm diameter stainless-steel pipe } with a flat top of 10mm thick polycarbonate. } } I've never liked having my face too close to the glass one when it's } in use, anyway. } } Anyone got any comments/recommendations? } } Anyone ever had a glass one implode in use? } } cheers } } rtch } } } Dr Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
There are two sources of noise, externally and internally generated. You have specificcally asked about echos so I assume that your problems stem from the rotary pumps, cooling fans, etc. on equipment that is essential to have operating in the room.
Try to get the rotary pumps and power supply rack into an area that can be screened off from the instrument column but remember that it will still need air flow for cooling.
For the desk and computer fans use some blackout curtains around the walls. This is readily available and is usually black one side and coloured the other so you can have a nice Oxford blue showing into the room:-) Make sure that it is at least 1.5 lengths, preferably 2x - ie. 7.5m of curtaining is a minimum for a 5m wall 10m is best. If you get the rotary pump and power supply at the back of the room you can hide them behind 3 layers of the same curtaining.
We have found this very effective for our HREM instruments and our FEGTEM.
Noise from outside the room needs dealing with outside the room.
Regards, Ron
} Dear Listers, } } Hope you are not knocked off from your normal life by the tragic events. } } Anyway, I have a question for you gurus: we have a 2010F TEM sitting in a } regular sized room. Somehow we feel there is a bit above normal acoustic } echos going on in the lab which interferes with the TEM. So we hope to } find some kind of insulation to be put on the wall and/or around the TEM to } improve the performance. } } Any thoughts and suggestions are highly appreciated. } } Thanks and take care. } } ********************************* } Chaoying Ni } Electron Microscopy Facility } College of Engineering } University of Delaware } Newark, DE 19716 } ********************************* } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
We have a 2010F in a similar sized room and we use floor to ceiling black curtaining - the heavier the better (ours are double thickness and lined) - and it makes a big difference. Hope this helps Alan Walker
********************************************* Alan Walker Dept of Electronic and Electrical Engineering University of Sheffield Mappin Street Sheffield S1 3JD United Kingdom
Don't use a polycarbonate end plate to cover the end of your vacuum chamber. A flat plate of polycarbonate isn't strong enough to span the opening.
Edwards did sell glass cylinders with metal endplates on some of their 306 coaters--in particular their old ion beam sputtering system. The unit I have here has a 1/2 inch thick Aluminum plate spanning a 12 inch diameter glass vacuum cylinder. The glass cylinder is the same type of glass you find on a normal bell jar. It just has an L-gasket on both the top and bottom. Be careful that you use an adequate grade of aluminum for the top plate.
The advantage of the cylinder with metal top plate is that you can have feedthroughs to the vacuum chamber from the top as well as through the base plate.
I suspect that you can get the glass cylindrical glass vacuum vessels and L-gaskets from the usual vacuum supply houses.
cheers, Henk Colijn
At 12:32 PM 9/19/01 +0000, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://www.ceof.ohio-state.edu Fools are pleased when they discover error. The wise are pleased when they discover truth.
I have not known a glass tube to implode but we use an outer cover just in case (perspex tube). Tubes are checked for cracks before use and previous problems have been obvious, usually by the many bits all over the floor!
My worry would be the 10mm polycarbonate top, at 300mm dia that's over 1500 pounds (750Kg?) force. I would want something thicker or stronger.
I also like being able to look in the side to see my coating set up after evacuation and the evaporation (through appropriate goggles).
Regards, Ron
} I've just had the misfortune to smash the glass bell jar on my } Edwards 306 coater. About 305mm dia, 400mm high. } } While I can and maybe should buy a glass replacement, I'm tempted to } make one from a 400mm length of 305mm diameter stainless-steel pipe } with a flat top of 10mm thick polycarbonate. } } I've never liked having my face too close to the glass one when it's } in use, anyway. } } Anyone got any comments/recommendations? } } Anyone ever had a glass one implode in use? } } cheers }
Dear Ritchie,
I have a polycarbonate lid for a vacuum apparatus with the same diameter as your coater. It is about 45 mm thick, and it has not given me any trouble. I agree with Ron that 10 mm is not thick enough.
Yours,
Bill Tivol Wadsworth Center Albany NY (518) 473-7399 WFT02-at-health.state.ny.us
We've had to contend with noisy rooms in nearly all of our electron microscope laboratories. Here is a list of things to do:
* Try to eliminate the source of the noise. * Call in the microscope manufacturer to provide a noise spectrum (dB versus Hz). Ask which frequencies are a problem (usually low frequencies). * Look for devices and materials which will attenuate the noise. Pay particular attention to their effectiveness in attenuating the problem frequencies.
Here are a few companies which manufacture items you may need: * Kinetics Noise Control, http://www.kineticsnoise.com * Industrial Acoustics Company, http://www.industrialacoustics.com * Industrial Noise Control, http://www.industrialnoisecontrol.com
I've worked several times with David Mitchell, VP at The Huff Co., Lake Bluff, IL (Chicago area), 1-847-362-7440, to ameliorate our noise problems. Such companies will evaluate the site, recommend solutions, and even install the various devices and materials from the manufacturers.
Some of the things we've done are: * glued "egg-crate" foam on the walls * installed fabric-covered fiberglass panels on the walls * installed duct silencers in air ducts * wrapped air ducts with fiberglass batting backed with "loaded vinyl" * hung fiberglass noise-absorbing curtains * installed isolation pads under vibrating equipment
If you install wall panels or foam, it is usually not necessary to cover all of the wall surfaces. There are lots of things to consider, so evaluate your site carefully.
..Russ Cook ---------------------------------------- Russell E. Cook, Ph.D. Electron Microscopy Center Argonne National Laboratory Materials Science Division, Building 212 9700 South Cass Avenue Argonne, IL 60439-4838 (630)252-7194 FAX: (630)252-4289 recook-at-anl.gov
} -----------------------------------------------------------------------. } Dear Listers, } } Hope you are not knocked off from your normal life by the tragic events. } } Anyway, I have a question for you gurus: we have a 2010F TEM sitting in a } regular sized room. Somehow we feel there is a bit above normal acoustic } echos going on in the lab which interferes with the TEM. So we hope to } find some kind of insulation to be put on the wall and/or around the TEM to } improve the performance. } } Any thoughts and suggestions are highly appreciated. } } Thanks and take care. } } ********************************* } Chaoying Ni } Electron Microscopy Facility } College of Engineering } University of Delaware } Newark, DE 19716 } *********************************
Is it common to use mouse antibodies (monoclonals) on mouse tissue for immunogold localizations? Is a control using the secondary antibody (anti-mouse-IgG-gold-conjugate) alone sufficient to make sure that one is detecting specificly the primary antibody and not tissue-internal mouse antibodies? Does somebody know examples from the literature where people used mouse antibodies (monoclonals) on mouse tissue for immunogold localizations? I am a botanist, so until now I never ran into that sort of problems.
Thanks,
Stefan
°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°° Stefan Geimer Universität zu Köln Botanisches Institut Gyrhofstr. 15 50931 Köln Germany °°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°
I didn't get to the MSA meeting this summer, but I heard through the grapevine about Gatan's new UltraScan 1000 and saw their poster. I was very impressed with the 4K x 4k montage taken at 5k mag. It looked publishable.I'd be interested in hearing comments/impressions from anyone who a chance to use it. Thanks,
Hank Adams Integrated Microscopy Core Molecular and Cellular Biology Baylor College of Medicine Houston, Tx 77030
We are about to install a Tecnai 20 TEM. In addition to isolating rotary pumps and power supply as suggested by Ron Doole, we are covering the walls with Armstrong Sound Soak Panels.
Frank
At 11:25 AM 9/18/01 -0400, Chaoying Ni wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Is it common to use mouse antibodies (monoclonals) on mouse tissue for immunogold localizations? Is a control using the secondary antibody (anti-mouse-IgG-gold-conjugate) alone sufficient to make sure that one is detecting specificly the primary antibody and not tissue-internal mouse antibodies? Or are there other things I can doo as a good control?
Does somebody know examples from the literature where people used mouse antibodies (monoclonals) on mouse tissue for immunogold localizations? I am a botanist, so until now I never ran into that sort of problems.
Thanks,
Stefan
°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°° Stefan Geimer Universität zu Köln Botanisches Institut Gyrhofstr. 15 50931 Köln Germany °°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°°
Hello to the person from down under with the broken bell jar:
I have been using a Denton 502A carbon evaporator for the past decade and have also had some visions of it imploding. The model I have has a metal mesh cover over the jar and while it would not stop little pieces of flying glass it would catch the big ones. As far as using a home made unit goes, I would be vary cautious. At the very least consult with some kind of structural engineer who is familiar with vacuum systems. Be careful. Tim
Timothy G. Schneider Director of Electron Microscopy Department of Pathology Thomas Jefferson University Room 229 JAH 1020 Locust Street Philadelphia Pa. 19107 215-503-4798 work 610-613-8170 cell Timothy.Schneider-at-Mail.TJU.edu
Sara, actually the total package for the UltraScan 1000 is about $100K. The UltraScan 4000 is the one you've quoted and that is a 4kx4k camera. The 1000 has the 2kx2k camera and montages the field requiring 4 exposures. I believe that is the way it works.
Hank
-----Original Message----- } From: Sara Miller [mailto:saram-at-duke.edu] Sent: Wednesday, September 19, 2001 1:58 PM To: Hank Adams
Dear Chaoying, One unlikely source of noise we recently found came from the diffusion pump on our CM 300. It was crackling like diffusion pumps due, but we could see our HRTEM image shake with the big crackles. The solution was deduced by our excellent FEI field engineer. It was to reduce the temperature in the diffusion pump by reducing the amount of oil charge. If you have noise problems that go snap, crackle, pop in the dark, try checking your oil. Ciao for now, Ken
Dear Chaoying, One unlikely source of noise we recently found came from the diffusion pump on our CM 300. It was crackling like diffusion pumps due, but we could see our HRTEM image shake with the big crackles. The solution was deduced by our excellent FEI field engineer. It was to reduce the temperature in the diffusion pump by reducing the amount of oil charge. If you have noise problems that go snap, crackle, pop in the dark, try checking your oil. Ciao for now, Ken
10 mm polycarbonate? I would suggest much thicker. Have no idea how much thick. I think the problem is that your pipe has a huge diameter. The force on your polycarbonate will be proportional to the size of area (1 kg/cm2 x 706 cm2=approx 700 kg, 1600 lb). I afraid, the total cost of the pipe etc would be comparable to the cost of Bell Jar. You may find Bell Jar on used equipment sale for about $300. Sergey
At 12:32 PM 9/19/01 +0000, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
This is incredible. I would suggest that this is a first EM digital camera which produces comparable to the film image quality. The chip is huge, so they have 80-90% area of the film on the chip. The images are very sharp. The technical problem there was to read information from such huge chip. They used new technology when chip (virtually?) splitted on quarters and read in parallel. They did a great job. The camera is very expensive by the way.
Sergey
At 11:00 AM 9/19/01 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
I think you wrong. They do have two new cameras 2x2K and 4x4K. This is real number of pixels on the chip. I also believe, Ultrascan1000 is much, much more expensive than you suggest. As per our conversation with Gatan people, they expect to start selling that cameras at the end of the year, so it's very unlikely the price is known for now (I did not ask them directly about the price, but have impression that it's much more expensive that previous models, which were in the $70K range). Current Gatan's cameras (600W/CW Multiscan for instance) already has capacity for multiple exposure I believe. Actually, it's not a properties of camera but software. It's called "Auto-montage" and you could "montage" as many pictures as you want. By the way, Multiscan cameras has 2-4x (not sure in exact number) bigger area of view than film, which sometime is very useful. Over last few month I was studying EM digital cameras on the makket very hard, so if you have questions, may be I could help.
Sergey.
At 02:42 PM 9/19/01 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
I would appreciate any advice on the following question. I am planning to do fluorescence microscopy and electron microscopy on the same sample. I would like to do immunolabelling on cells grown in a culture dish followed by plastic embedding of the same sample and further preparation for EM. In order to find the right area of cells to be sectioned for EM (previously detected by immuno fluorescence) I suppose one has to have some kind of grid on the culture dish. I am aware of the fact that there are gridded coverslips available, however, I always had a lot of trouble to remove the coverslip from the glass after embedding (used lN2 and so on). So it would be very helpful if anyone out there could provide any other working recipes, references for methods or sources for culture dishes with some kind of grid on them.
Dr. Andreas Brech EM-Unit, Dept. of Biology, University of Oslo. P.O.Box 1062 Blindern, Room U 139 Street address: Moltke Moes vei 32 N-0316 Oslo 3 Norway Tel.: + 47-22 85 47 25/61 89 (work) + 47-22 43 83 23 (privat) Fax.: + 47-22 85 47 26 e-mail.: abrech-at-bio.uio.no
I have a visitor who wants to use acridine orange (0.1%) in seawater (or artificial seawater) to stain nuclei/DNA in limpet eggs and sperm during fertilisation. It is done by other workers, but so far we've found no information as to how long it takes to stain things. Any suggestions? Any pH considerations?
Also, does it work on fresh (i.e. marine, seawater)and formalin-fixed material with equal efficiency.
Finally, we understand that the fluorescence is short-lived - any comments as to how long a time period would be helpful.
Many thanks
Keith Ryan Marine Biological Association & University of Plymouth UK
The three-day intensive hands-on workshop on Image Processing and Measurement presented by John Russ through the North Carolina State University Department of Continuing and Professional Education is now in its 19th year. The course will be presented October 30 - November 1, in Raleigh, NC. This course has generated highly favorable reviews from the thousands of previous students. The primary focus is on images from various types of microscopy, with practical guidance in correcting imaging defects, enhancing the images for presentation and measurement, and performing stereological meaningful measurements on them. Textbooks and computer software are provided to attendees. Lab sessions with an opportunity to bring your own images makes this course immediately useful and highly productive.
For full information on the course, including outlines, faculty information, a downloadable brochure, and on-line registration, go to
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or call Cindy Allen, 919-515-8171, at North Carolina State University Dept. of Continuing and Professional Education.
I am gathering data on enviornmental SEMs. Hopefully I'll be setting up a research facility within the coming months.
Does anyone have preference to systems?
What, if any differences are there for maintinence compared to a standard SEM?
COST? (ouch)
Thanks,
Tim Quinn University of Kansas Natural History Museum & Biodiversity Research Center 1345 Jayhawk Blvd 414A Lawrence, KS 66045 785-864-4556 tquinn-at-ku.edu
That *is* incredible. Chip has length of 4080 pixels by 15 micron pixel pitch = 61.2 mm. Area is then 37.45 square cm or 5.8 square inches. Plate (film) size is 3.25 inches by 4.00 inches = 13 square inches. Chip area is 45% of film area... -Mike
Sergey Ryazantsev wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } This is incredible. I would suggest that this is a first EM digital camera } which produces comparable to the film image quality. The chip is huge, so } they have 80-90% area of the film on the chip. The images are very sharp. } The technical problem there was to read information from such huge } chip. They used new technology when chip (virtually?) splitted on quarters } and read in parallel. They did a great job. The camera is very expensive } by the way. } } Sergey } } At 11:00 AM 9/19/01 -0500, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } I didn't get to the MSA meeting this summer, but I heard through the } } grapevine about Gatan's new UltraScan 1000 and saw their poster. I was very } } impressed with the 4K x 4k montage taken at 5k mag. It looked } } publishable.I'd be interested in hearing comments/impressions from anyone } } who a chance to use it. } } Thanks, } } } } Hank Adams } } Integrated Microscopy Core } } Molecular and Cellular Biology } } Baylor College of Medicine } } Houston, Tx 77030 } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } Box 951737 } Los Angeles, CA 90095-1737 } } Phone: (310) 825-1144 } Pager: (310) 845-0248 } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu
The New York Society of Experimental Microscopists Announcing the 50th Anniversary Presidential Symposium
"FRONTIERS in MICROSCOPY"
Caspary Auditorium, Rockefeller University October 11, 2001 9:00 AM-4:00 PM Free and open to the Public
50 Years of NYSEM C. DeLemos-Chiarandini, New York University
George Palade, and the Early Days of Electron Microscopy in New York P. Satir, Albert Einstein College of Medicine
EM Tomography Reveals Cell Structure at 6nm Resolution J. R. McIntosh, University of Colorado
Multiphoton Imaging of the Molecular Dynamics of Life Processes W.Webb, Cornell University
Intravital Imaging of Tumor Cells J.Condeelis, Albert Einstein College of Medicine
Total Internal Reflectance Microscopy Visualizes Membrane Fusion Events S. Simon, Rockefeller University
Imaging Biochemistry Inside Cells P. Bastiaens, EMBL, Heidelberg
The Ribosome: Snapshots of a Molecular Machine in Motion J. Frank, HHMI, Wadsworth Center
Imaging RNA in Living Cells R. Singer, Albert Einstein College of Medicine
High Temporal and Spectral Resolution using Acousto-Optical Tunable Filters D. Farkas, Carnegie-Mellon University
**************************************************************************** Frank Macaluso tel: 718-430-3547 Analytical Imaging Facility fax: 718-430-8996 Albert Einstein College of Medicine e-mail: macaluso-at-aecom.yu.edu 1300 Morris Park Avenue http://www.aecom.yu.edu/aif/ Bronx, NY 10461 ****************************************************************************
You can culture your cells on plastic coverslips and embed them in resin. The embedded cells are good for both IFM and iEM but you have to work out the conditions. The coverslip is called Thermanox from EMS.
Greg
Gang Ning, M.D., Ph.D. Electron Microscopy Facility Medical College of Wisconsin
Andreas Brech wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I would appreciate any advice on the following question. I am planning to do } fluorescence microscopy and electron microscopy on the same sample. I would } like to do immunolabelling on cells grown in a culture dish followed by plastic } embedding of the same sample and further preparation for EM. } In order to find the right area of cells to be sectioned for EM (previously } detected by immuno fluorescence) I suppose one has to have some kind of grid } on the culture dish. I am aware of the fact that there are gridded coverslips } available, however, I always had a lot of trouble to remove the coverslip from } the glass after embedding (used lN2 and so on). So it would be very helpful if } anyone out there could provide any other working recipes, references for } methods or sources for culture dishes with some kind of grid on them. } } Dr. Andreas Brech } EM-Unit, Dept. of Biology, } University of Oslo. } P.O.Box 1062 Blindern, Room U 139 } Street address: Moltke Moes vei 32 } N-0316 Oslo 3 } Norway } Tel.: + 47-22 85 47 25/61 89 (work) } + 47-22 43 83 23 (privat) } Fax.: + 47-22 85 47 26 } e-mail.: abrech-at-bio.uio.no
Hi Listers, I have a question about a schottky tipped FE Microscope. Specifically, what is the best way to shut the beam off at the end of the day. We will be using probably everyday for long sessions, so would beam blanking be preferable to shutting off the extraction voltage. I am wondering about the effects of tip life, beam stability, and contamination. Any advice or past experience would be greatly appreciated. Thanks again Nick Aitken
Nicol Aitken Sample Preparation and Imaging Specialist Research and Development Semiconductor Insights Inc. email:nicol-at-semiconductor.com (613)599-6500 ext.4300
Yes, I did check your WEB-site in the beginning of my search a few month ago as well as many others. My initial criteria were the following:
-35 mm port mount; -price; -ability to have demo for the camera; -ability to have reasonable service in time.
You product was excluded from initial list for the following reasons:
- your Biocam camera, which was possible candidate is bottom-mounted;
- on your WEB-site at http://www.tvips.com/Contact_en.htm I did not find any information about your headquarter in US or contact phones, so I was not able to talk to your representative (I was using the telephone call as a first "try" of the customer service quality: if I was on hold for more than 5 min, I excluded the candidate from my list).
-location of your head office in Germany makes me skeptical about customer service quality company could provide in US.
I would like to mention that my criteria is subjective and based on my own way to make a business and spent my money. My decision is not related to the real quality of your products.
I would mention also that a few companies refused to give me demo on their cameras (or delayed with answer) and were excluded from the list as well.
Sergey.
At 02:30 PM 9/20/01 +0200, you wrote: } Dear Mr. Sergey Ryazantsev, } } I have read in your email, that you have studied cameras for TEM. } Have you studied our cameras as well? } } For example, this year we have had the following installations in California: } - Scripps in San Diego, F-224 at a CM-200FEG (Ron Milligan's lab) } - UC in San Diego, F-224/FastScan at a JEM-200EX (Mark Ellisman's lab) } - Quantum Dot Corporation in Hayward, TemCam-0124 at JEM-200CX } - Additionally just this month Bridget Carragher (together with her group) has } moved from Illinois to the Scripps Institute. She will mount another F-224 } at a } Tecnai-20FEG. } } If you want to know more about our US installations and our cameras, } please let } me know. } } Best wishes, } Ingo Daberkow } } BTW: We are in contact with Phoebe Stewart from the Department of Molecular & } Medical Pharmacology, UCLA School of Medicine } } +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ } Dr. Ingo Daberkow } Tietz Video and Image Processing Systems GmbH } Herbststrasse 7 } D-82131 Gauting, Germany } Tel: +49-89-8506567 } FAX: +49-89-8509488 } Internet: http://www.tvips.com/ } Email: ingo.daberkow-at-tvips.com } +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ } } } Sergey Ryazantsev wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } Hank, } } } } I think you wrong. They do have two new cameras 2x2K and 4x4K. This is } } real number of pixels on the chip. I also believe, Ultrascan1000 is much, } } much more expensive than you suggest. As per our conversation with Gatan } } people, they expect to start selling that cameras at the end of the year, } } so it's very unlikely the price is known for now (I did not ask them } } directly about the price, but have impression that it's much more expensive } } that previous models, which were in the $70K range). Current Gatan's } } cameras (600W/CW Multiscan for instance) already has capacity for multiple } } exposure I believe. Actually, it's not a properties of camera but } } software. It's called "Auto-montage" and you could "montage" as many } } pictures as you want. By the way, Multiscan cameras has 2-4x (not sure in } } exact number) bigger area of view than film, which sometime is very } } useful. Over last few month I was studying EM digital cameras on the } } makket very hard, so if you have questions, may be I could help. } } } } Sergey. } } } } At 02:42 PM 9/19/01 -0500, you wrote: } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } Sara, actually the total package for the UltraScan 1000 is about } $100K. The } } } UltraScan 4000 is the one you've quoted and that is a 4kx4k camera. } The 1000 } } } has the 2kx2k camera and montages the field requiring 4 exposures. I } believe } } } that is the way it works. } } } } } } Hank } } } } } } -----Original Message----- } } } } From: Sara Miller [mailto:saram-at-duke.edu] } } } Sent: Wednesday, September 19, 2001 1:58 PM } } } To: Hank Adams } } } Subject: Re: Gatan's new UltraScan 1000 } } } } } } } } } Micrograph was lovely. Price tag was in range of $275 K--ouch! They } didn't } } } have one there on demo that I saw. } } } } } } Sara E. Miller, Ph. D. } } } P. O. Box 3712 } } } Duke University Medical Center } } } Durham, NC 27710 } } } Ph: 919 684-3452 } } } FAX: 919 684-3265 } } } } _____________________________________ } } } } Sergey Ryazantsev Ph. D. } } Electron Microscopy } } UCLA School of Medicine } } Department of Biological Chemistry } } Box 951737 } } Los Angeles, CA 90095-1737 } } } } Phone: (310) 825-1144 } } Pager: (310) 845-0248 } } FAX (departmental): (310) 206-5272 } } mailto:sryazant-at-ucla.edu
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Steve Pfeiffer of Gatan just E.mail to me that price range for Ultrascan 1000 is about $100K. Which is much less than I expect to see for such camera.
Sergey
At 10:06 AM 9/20/01 -0500, you wrote: } Sergey, I just talked to Gatan about their UltraScan 1000 that tiles 4 } images together. The entire package with installation/training and the } Automontage plug-in minus the computer/monitor is about $100K. Using their } specs for the computer add another $4800 which includes a 21" monitor. The } CCD magnifies the image 1.3x-1.5x compared to film. } } Hank } -----Original Message----- } From: Sergey Ryazantsev [mailto:sryazant-at-ucla.edu] } Sent: Wednesday, September 19, 2001 8:10 PM } To: Hank Adams; Microscopy-at-sparc5.microscopy.com } Subject: RE: Gatan's new UltraScan 1000 } } } Hank, } } I think you wrong. They do have two new cameras 2x2K and 4x4K. This is } real number of pixels on the chip. I also believe, Ultrascan1000 is much, } much more expensive than you suggest. As per our conversation with Gatan } people, they expect to start selling that cameras at the end of the year, } so it's very unlikely the price is known for now (I did not ask them } directly about the price, but have impression that it's much more expensive } that previous models, which were in the $70K range). Current Gatan's } cameras (600W/CW Multiscan for instance) already has capacity for multiple } exposure I believe. Actually, it's not a properties of camera but } software. It's called "Auto-montage" and you could "montage" as many } pictures as you want. By the way, Multiscan cameras has 2-4x (not sure in } exact number) bigger area of view than film, which sometime is very } useful. Over last few month I was studying EM digital cameras on the } makket very hard, so if you have questions, may be I could help. } } Sergey. } } At 02:42 PM 9/19/01 -0500, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Sara, actually the total package for the UltraScan 1000 is about $100K. The } } UltraScan 4000 is the one you've quoted and that is a 4kx4k camera. The } 1000 } } has the 2kx2k camera and montages the field requiring 4 exposures. I } believe } } that is the way it works. } } } } Hank } } } } -----Original Message----- } } } From: Sara Miller [mailto:saram-at-duke.edu] } } Sent: Wednesday, September 19, 2001 1:58 PM } } To: Hank Adams } } Subject: Re: Gatan's new UltraScan 1000 } } } } } } Micrograph was lovely. Price tag was in range of $275 K--ouch! They } didn't } } have one there on demo that I saw. } } } } Sara E. Miller, Ph. D. } } P. O. Box 3712 } } Duke University Medical Center } } Durham, NC 27710 } } Ph: 919 684-3452 } } FAX: 919 684-3265 } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } Box 951737 } Los Angeles, CA 90095-1737 } } Phone: (310) 825-1144 } Pager: (310) 845-0248 } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
For the past 4 years we have run the thermal emitter in our Zeiss LEO DSM982 almost everyday Monday-Friday and off on weekends and vacations. The previous emitter (Zeiss OEM) lasted 14,900 hours over 3 years. The present emitter (from FEI) has 3,000 hours since November 2000. We routinely image at } 100kx with great results. What microscope do you have?
Jim
Nicol Aitken wrote:
Hi Listers, I have a question about a schottky tipped FE Microscope. Specifically, what is the best way to shut the beam off at the end of the day. We will be using probably everyday for long sessions, so would beam blanking be preferable to shutting off the extraction voltage. I am wondering about the effects of tip life, beam stability, and contamination. Any advice or past experience would be greatly appreciated. Thanks again Nick Aitken
Nicol Aitken Sample Preparation and Imaging Specialist Research and Development Semiconductor Insights Inc. email:nicol-at-semiconductor.com (613)599-6500 ext.4300
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility Unit-2131 Storrs, CT 06269-2131 bsgphy3-at-uconnvm.uconn.edu 860 486-2914 voice 860 486-1936 fax
There may be a difference in TEM and SEM FEG use that I am not familiar with, I am primarily a TEM guy, however we keep our emission running as much as possible.
With our JEOL3000F FEGTEM we run the tip down maybe 4 times a year, other than that it is on and the valve between the gun and the column is closed when the instrument is not being used. There are several reasons for this.
It takes a couple of hours to run up and condition the HT at 300kV and then another couple of hours to run up the emission. This makes it impossible to switch on the emission for a day's use.
Having run up the emission it is not stable, the emission current keeps dropping. This exponential decrease takes several hours to stabilise (we allow 6 hours before we think it is really stable). Of course if you don't need a stable current (eg. for HREM imaging) you can still use it.
The most likely time for any damage to the tip (apart fom accidents) is when it is heated or cooled so we don't want to do that more than neccessary.
I guess that there is finite life of the tip so it would be pointless to keep it running for a long time if the instrument is not being used. Does anyone know why a tip should eventually fail if operating conditions are optimised?
Our present tip (the first) has been running during factory tests and since installation in Oxford in 1998 and has run for over 21,500 hours.
Regards, Ron ps. If our tip now fails I'm going to blame you guys for raising the subject.
} Nicol, } } For the past 4 years we have run the thermal emitter in our Zeiss LEO } DSM982 almost everyday Monday-Friday and off on weekends and vacations. The } previous emitter (Zeiss OEM) lasted 14,900 hours over 3 years. The present } emitter (from FEI) has 3,000 hours since November 2000. We routinely image } at } 100kx with great results. } What microscope do you have? } } Jim } } } Hi Listers, } I have a question about a schottky tipped FE Microscope. Specifically, what } is the best way to shut the beam off at the end of the day. We will be using } probably everyday for long sessions, so would beam blanking be preferable to } shutting off the extraction voltage. I am wondering about the effects of tip } life, beam stability, and contamination. Any advice or past experience would } be greatly appreciated. } Thanks again } Nick Aitken } } Nicol Aitken } Sample Preparation and Imaging Specialist } Research and Development } Semiconductor Insights Inc. } email:nicol-at-semiconductor.com } (613)599-6500 ext.4300 } } James S. Romanow } The University of Connecticut } Physiology and Neurobiology Department } Electron Microscopy Facility } Unit-2131 } Storrs, CT 06269-2131 } bsgphy3-at-uconnvm.uconn.edu } 860 486-2914 voice } 860 486-1936 fax } } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
For the most complete information on fluorescence probes (which you may already know), try URL: http://www.probes.com/; Search: "Acridine Orange" for a plethora (my son accused me of a "plethora of harassment", the other night!) of information and citations.
You might also consult Hayat, MA, Stains and Cytochemical Methods, Plenum, 1993, ISBN: 0-306-44294-9 for an informative, though not explicit, description of AO and its applications and properties.
For the reason that AO has been used extensively in flow cytometry, you should also look into the several chapters that are relevant to your question in: Methods in Cell Biology, vol 33, Flow Cytometry, Darzynkiewicz and Crissman(Eds.)Academic Press, 1990, ISBN: 0-12-564133-8 or 0-12-203050-8.
pH is important to the fluorescent properties of the dissolved dye, but I have not observed such effects in my experience which has been confined to viewing virus-infected cultured cells (vital, but usually fixed/dead). My original histochemical use of AO fluorescence - early '60's - was to distinguish between DNA and RNA - until it was pointed out by virologists that there were some single-stranded DNA viruses that could interfere with interpretation. Now we know that AO can be used to intercalate preferentially in transcribing DNA, thus providing an indication of RNA production.
I hope this helps, and
Good luck,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging(CASI) West Chester University of Pennsylvania Schmucker Science Center II South Church Street West Chester, PA, 19383 eMail: fmonson-at-wcupa.edu http://darwin.wcupa.edu/casi/
} ---------- } From: Keith Ryan } Reply To: kpr-at-mba.ac.uk } Sent: Thursday, September 20, 2001 7:47 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: LM - acridine orange - help } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Hello Listers } } I have a visitor who wants to use acridine orange (0.1%) in seawater (or } artificial seawater) to stain nuclei/DNA in limpet eggs and sperm during } fertilisation. It is done by other workers, but so far we've found no } information as to how long it takes to stain things. Any suggestions? Any } pH considerations? } } Also, does it work on fresh (i.e. marine, seawater)and formalin-fixed } material with equal efficiency. } } Finally, we understand that the fluorescence is short-lived - any comments } as to how long a time period would be helpful. } } } Many thanks } } Keith Ryan } Marine Biological Association } & University of Plymouth } UK } } PLymouth } } }
my experience is the same as jim's. i've got about 10Khours on an fei emitter now...still works great at } 500KX. its "on" all week and "off" most weekends.
b-
} X-Comment: UConnVM.UConn.Edu: Mail was sent by d40h44.public.uconn.edu } Date: Thu, 20 Sep 2001 17:59:41 -0400 } To: Microscopy-at-sparc5.microscopy.com } From: Jim Romanow {bsgphy3-at-uconnvm.uconn.edu} } Subject: Thermal FE } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thanks for the fast reaction, *but* you have forwarded my direct email to you to the listserver as well.
Do you really want to discuss this matter at the listserver?
Of course, I am willing to discuss the advantage/disadvantage of our cameras in publicity, buy I fear this might be regarded as advertisement!?
Therefore, at first, I want to know the opinion of the list server community! Nestor, maybe do you can comment it?
Best wishes, Ingo
Sergey Ryazantsev wrote:
} Dear Ingo } } Yes, I did check your WEB-site in the beginning of my search a few month } ago as well as many others. My initial criteria were the following: } } -35 mm port mount; } -price; } -ability to have demo for the camera; } -ability to have reasonable service in time. } } You product was excluded from initial list for the following reasons: } } - your Biocam camera, which was possible candidate is bottom-mounted; } } - on your WEB-site at http://www.tvips.com/Contact_en.htm I did not find } any information about your headquarter in US or contact phones, so I was } not able to talk to your representative (I was using the telephone call as } a first "try" of the customer service quality: if I was on hold for more } than 5 min, I excluded the candidate from my list). } } -location of your head office in Germany makes me skeptical about customer } service quality company could provide in US. } } I would like to mention that my criteria is subjective and based on my own } way to make a business and spent my money. My decision is not related to } the real quality of your products. } } I would mention also that a few companies refused to give me demo on their } cameras (or delayed with answer) and were excluded from the list as well. } } Sergey. }
Only the FEI ESEM allows you to have water in the chamber. If I had the money I would get the FEGESEM.
Re maintainance, we do not have any extra problems with our XL30 ESEM as our wet bullet apertures are replaced at service. Our contract gives us 4 services a year. It would be useful to get comments from a user without a service contract.
Dave
On Thu, 20 Sep 2001 10:37:47 -0500 "Quinn, Tim Lee" {tquinn-at-ku.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Listservers, } } I am gathering data on enviornmental SEMs. Hopefully I'll be setting up a } research facility within the coming months. } } Does anyone have preference to systems? } } What, if any differences are there for maintinence compared to a standard } SEM? } } COST? (ouch) } } Thanks, } } Tim Quinn } University of Kansas } Natural History Museum & } Biodiversity Research Center } 1345 Jayhawk Blvd 414A } Lawrence, KS 66045 } 785-864-4556 } tquinn-at-ku.edu }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
I am interested in subscribing to the microscopy listserver.
I have a question about immunogold labeling. I am interested in surface labeling receptors and examining patterns of internalization. Has anyone tried to label the surface receptor with a primary antibody in the live cell, induce internalization, fix and embed the cells. Then thin section the preparation and do postembedding with a secondary antibody tagged with a gold particle? Do you know if the primary antibody will survive the embedding process with low-termperature embedding with lowicryl? Thanks for your help. Michelle Adams
Hi All, I hope to be installing a new TEM soon and unfortunately have some concerns about magnetic field interference with the column.
Being a new "immigrant" to the USA I am not up to speed with vendors on this side of the Atlantic. I need to know who supplies field canceling systems in the USA - calls from vendors welcome. I would also be interested in colleagues experiences with individual systems. I may have to deal with particularly high field strengths in the vicinity of the microscope so would be interested in what is the maximum field strength that such systems can compensate. Another alternative would be to look into Mu metal lining for the microscope room. Again info on vendors would be most useful as well as anyone's experience of using this method of shielding.
Many thanks
Chris
Christopher Gilpin Ph.D. Assistant Professor Director, Molecular and Cellular Imaging Facility Cell Biology Department University of Texas Southwestern Medical Center at Dallas Dallas, TX, 75390-9039 phone +1 214 648 2827 fax +1 214 648 6408 christopher.gilpin-at-utsouthwestern.edu
On a related note we just purchased a camera from Ingo. We have been extremely happy with the performance. The camera was installed about one mouth after tvips received the purchase order and the instating went very smoothly. If anyone has questions please contact me.
Glenn
} Sergey, } } Thanks for the fast reaction, *but* you have forwarded my direct email to } you to } the listserver as well. } } Do you really want to discuss this matter at the listserver? } } Of course, I am willing to discuss the advantage/disadvantage of our } cameras in } publicity, buy I fear this might be regarded as advertisement!? } } Therefore, at first, I want to know the opinion of the list server community! } Nestor, maybe do you can comment it? } } Best wishes, } Ingo } } } Sergey Ryazantsev wrote: } } } Dear Ingo } } } } Yes, I did check your WEB-site in the beginning of my search a few month } } ago as well as many others. My initial criteria were the following: } } } } -35 mm port mount; } } -price; } } -ability to have demo for the camera; } } -ability to have reasonable service in time. } } } } You product was excluded from initial list for the following reasons: } } } } - your Biocam camera, which was possible candidate is bottom-mounted; } } } } - on your WEB-site at http://www.tvips.com/Contact_en.htm I did not find } } any information about your headquarter in US or contact phones, so I was } } not able to talk to your representative (I was using the telephone call as } } a first "try" of the customer service quality: if I was on hold for more } } than 5 min, I excluded the candidate from my list). } } } } -location of your head office in Germany makes me skeptical about customer } } service quality company could provide in US. } } } } I would like to mention that my criteria is subjective and based on my own } } way to make a business and spent my money. My decision is not related to } } the real quality of your products. } } } } I would mention also that a few companies refused to give me demo on their } } cameras (or delayed with answer) and were excluded from the list as well. } } } } Sergey. } }
Glenn Fried Imaging Technology Group Beckman Institute University of Illinois phone 217 333 5493 Fax 217 244 6219
I shut extraction voltage off only when I was going on a long vacation and when I was told there could be a possibility of power outage (I put a microscope in a stand-by mode). Thermo cycling is the second worst thing you can do to the tip after power outage.
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: Nicol Aitken [mailto:nicol-at-semiconductor.com] } Sent: Thursday, September 20, 2001 3:09 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Thermal FE } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } } Hi Listers, } I have a question about a schottky tipped FE Microscope. } Specifically, what } is the best way to shut the beam off at the end of the day. } We will be using } probably everyday for long sessions, so would beam blanking } be preferable to } shutting off the extraction voltage. I am wondering about the } effects of tip } life, beam stability, and contamination. Any advice or past } experience would } be greatly appreciated. } Thanks again } Nick Aitken } } Nicol Aitken } Sample Preparation and Imaging Specialist } Research and Development } Semiconductor Insights Inc. } email:nicol-at-semiconductor.com } (613)599-6500 ext.4300 } } }
I have been getting mail from root-at-mail.advancelink.com all afternoon. Twenty or more of these emails. Other (I think that are on this list) have been getting the same email but from my email address. I have not sent these messages.
I have this bad feeling that this is a virus and it I have gotten from the listserver somehow. Could the admin of this listserver check this out.
What's the rationale for turning it off over weekends?
My system uses, I believe, a Denka ZrO/W filament in my Amray 305FE gun assembly. I never shut the gun off. The main problem with this particular SEM is that the gun valve is manual. If the gun is on and the gun valve is closed, there is most likely to be an air burp into the gun chamber and will poison the filament. This process also typically will burn the valves O-rings due to the high intensity FE emitter beam. So the valve is never closed until after turning off the filament.
The only reason I have found to worry about closing the gun valve is if there is a power failure. That would shut down the filament and all pumps (mech, turbo, column and gun ion pumps). I solved this by using two Toshiba 1400XL+ 6KVA dual conversion UPS units. Now, I shut the console off but keep it leaves the pumps and gun running. Plus, the UPS units
I log gun and column vacuum every few days along with extractor current. Then, about every month, I measure gun brightness. Running the way I am now, brightness is +- no more than about 0.4nA. Iext may change 3-6uA but brightness is way less affected by changes in Iext. When Iext and brightness start to drop or waver, that is an onerous sign that the gun is starting to fail.
I would tend to stick with this pattern since I know that bringing up a new gun takes about a week for the Iext and brightness to stabilize. I suspect that there would be some proportional time to stabilize if shut down over a weekend or for some extended time. Since I don't use the SEM every day, if shutting off the filament during some periods of time would extend its life, that is very desirable.
I have 9,020 hours on this gun since newly installed. It is very stable.
gary g.
At 05:51 AM 9/21/2001, you wrote:
} hi- } } my experience is the same as jim's. i've got about 10Khours on an fei } emitter now...still works great at } 500KX. its "on" all week and "off" } most weekends. } } b- } } } } X-Comment: UConnVM.UConn.Edu: Mail was sent by d40h44.public.uconn.edu } } Date: Thu, 20 Sep 2001 17:59:41 -0400 } } To: Microscopy-at-sparc5.microscopy.com } } From: Jim Romanow {bsgphy3-at-uconnvm.uconn.edu} } } Subject: Thermal FE } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Nicol, } } } } For the past 4 years we have run the thermal emitter in our Zeiss LEO } } DSM982 almost everyday Monday-Friday and off on weekends and vacations. The } } previous emitter (Zeiss OEM) lasted 14,900 hours over 3 years. The present } } emitter (from FEI) has 3,000 hours since November 2000. We routinely image } } at } 100kx with great results. } } What microscope do you have? } } } } Jim } } } } } } } } } } } } } } } } Nicol Aitken wrote: } } } } Hi Listers, } } I have a question about a schottky tipped FE Microscope. Specifically, what } } is the best way to shut the beam off at the end of the day. We will be using } } probably everyday for long sessions, so would beam blanking be preferable to } } shutting off the extraction voltage. I am wondering about the effects of tip } } life, beam stability, and contamination. Any advice or past experience would } } be greatly appreciated. } } Thanks again } } Nick Aitken } } } } Nicol Aitken } } Sample Preparation and Imaging Specialist } } Research and Development } } Semiconductor Insights Inc. } } email:nicol-at-semiconductor.com } } (613)599-6500 ext.4300 } } } } James S. Romanow } } The University of Connecticut } } Physiology and Neurobiology Department } } Electron Microscopy Facility } } Unit-2131 } } Storrs, CT 06269-2131 } } bsgphy3-at-uconnvm.uconn.edu } } 860 486-2914 voice } } 860 486-1936 fax } } } } } } } } } } } ---------------------------------------------- } Brian McIntyre } Electron Microscopy Lab- River Campus } Univ of Rochester } Rochester, NY 14627 } 716-275-3058/4875
I received this reply from a person about the cron daemon problem this afternoon. If anyone has continuing problems, I suggest you contact them directly.
--John chandler-at-colostate.edu Colorado State University
} From: "Kim M. Shiu" {kmshiu-at-mail.advancelink.com} } Subject: Re: Random mail from your server to me } To: chandler-at-lamar.colostate.edu (John Chandler) } Date: Fri, 21 Sep 101 16:23:52 -0700 (PDT) } MIME-Version: 1.0 } Status: RO } } Hi, } } There was a problem with a program on our server. It was not intentional } to send multiple copies or spam anyone. The problem has been corrected } and we apologize for any inconvenience it caused. } } Thanks. } } Kim Shiu } Advancelink } } } } } I sent this message to postmaster-at-advancelink.com and the messages continue } } to be sent. Please attend to this immediately. } } } } --John } } } } FYI, I have receive 7 messages from your mail server this afternoon similar } } or identical to the one below. This might indicate a situation that } } requires your attention. } } } } John Chandler } } Colorado State University } } chandler-at-lamar.colostate.edu } } } } } } } } ==============================================================================
} } } } } } Received: from mail.advancelink.com ([207.214.173.11]) by } } lamar.ColoState.EDU (AIX4.3/8.9.3/8.8.8) with ESMTP id NAA237230 for } } {john.chandler-at-colostate.edu} ; Fri, 21 Sep 2001 13:28:56 -0600 } } Received: (from root-at-localhost) } } by mail.advancelink.com (8.8.5/8.8.5) id LAA29356; } } Fri, 21 Sep 2001 11:35:00 -0700 (PDT) } } Date: Fri, 21 Sep 2001 11:35:00 -0700 (PDT) } } Message-Id: {200109211835.LAA29356-at-mail.advancelink.com} } } From: root-at-mail.advancelink.com (Cron Daemon) } } To: coccust-at-mail.advancelink.com } } Subject: Cron {coccust-at-mail} csh -x ~/bin/get-dist } get-dist.log } } X-Cron-Env: {SHELL=/bin/sh} } } X-Cron-Env: {HOME=/usr/home/coc/coccust} } } X-Cron-Env: {LOGNAME=coccust} } } X-Cron-Env: {USER=coccust} } } Status: } } } } cd /usr/home/coc/coccust } } grep [a-z] /usr/home/coc/cocinfo/MAILING-LISTS/CURRENT-LIST.txt } } chown coccust .forward } } chown: Command not found. } } chown coc .forward } } chown: Command not found. } } chmod 664 .forward } } exit } } } } } } }
Greetings Stefan, My initial reaction is that it is not common to use murine (mouse) monoclonal antibodies on mouse tissue. The reason for this is that if these murine antibody hybridomas were derived from mouse B-cells, and these B-cells produced antibodies to a mouse protein, one would expect an autoimmune response in the original mouse. In other words, it not be logical to produce an antibody against mouse proteins in a mouse because either: 1.) a secondary immune response would not be observed, or 2.) the ensuing autoimmune response would kill or at least severly debilitate the mouse. I guess recombinant monoclonal murine antibodies against mouse proteins could be produced however. A more likely scenario is that these mouse monoclonal antibodies are against a protein antigen derived from another species. A couple of important controls are in order. The simplest is to assess non-specific binding of your secondary (gold-conjugate) antibody. To do this you might block your tissue as usual, apply PBS or murine pre-immune sera to the tissue in place of the primary antibody, and then apply your secondary antibody. Another important control is to asses non-specific binding of your primary antibody. This is an isotype control. You should be able to find out the isotype of your monoclonal antibody from the source (eg. IgG K2B). Then obtain a monoclonal antibody of the same isotype which is not specific for any mouse protein (eg. anti-dansyl or something). This is used in place of the mouse specific monoclonal to assess non-specific binding due to charge properties. An isotype control for the secondary antibody could be conducted as well to assess the presence of tissue internal antibodies. -Karl G. _________________________________________________ Karl Garsha Light Microscopy Specialist Imaging Technology Group Beckman Institute for Advanced Science and Technology Room B650J University of Illinois at Urbana-Champaign Urbana, IL 61801 Tel: (217) 244-6292 Fax: (217) 244-6219 www.itg.uiuc.edu
Greetings Listers, As someone new to the challenge of managing multiuser equipment, I have come to realize that it will be important to devise an effective strategy to minimize wear and tear on instrumentation. It will also be important to attempt to try to salvage the results of the inevetable mistakes that users make. I am hoping some of the seasoned veterans out there can offer some advice. My question concerns how best to try to clean non-oil objectives which have been used with oil. I was previously under the impression that attempting this was futile, however I am hoping that I'm wrong. A collegue of mine suggested sonicating in acetone for a couple of minutes, but I'm hoping a gentler approach will suffice. Would the use o