We have been asked to look at the cell wall structure of some carrots and have essentially no experience in this area at all. At this stage we are interested in the cellulose structure to start with.
We have access to a Hitachi VP FESEM and JEOL TEM as well as a range of optical techniques. What would be the best technique for sample preparation (CPD etc) and imaging?
Thank you very much in advance for your help with this.
Colin Veitch
Electron Microscopist CSIRO Textile and Fibre Technology PO Box 21, BELMONT, Vic. 3216. Australia. colin.veitch-at-csiro.au http://www.tft.csiro.au
The information contained in this e-mail message may be privileged or confidential information. If you are not an intended recipient, you may not copy, distribute or take any action in reliance on it. If you have received this message in error, please telephone CSIRO Textile and Fibre Technology on +61 3 5246 4000.
==============================Original Headers============================== 10, 30 -- From Colin.Veitch-at-csiro.au Tue May 1 00:56:22 2007 10, 30 -- Received: from vic-MTAout3.csiro.au (vic-MTAout3.csiro.au [150.229.64.39]) 10, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l415uKdm015200 10, 30 -- for {microscopy-at-msa.microscopy.com} ; Tue, 1 May 2007 00:56:21 -0500 10, 30 -- DomainKey-Signature: s=email; d=csiro.au; c=nofws; q=dns; b=uZBrvaD0cxMV9MbIOdyLEoSa9e/31sIKDfJqM+mVQ4PGmU2m4Cex00xvVXHhJrzY5SufCwEgczAoIx1gjnxnvfVGJ+Vtlh8poqgarafPm4Vk8DsxYJn4bPGmCcNy6pjO; 10, 30 -- Received: from exgw1-mel.nexus.csiro.au ([138.194.3.56]) 10, 30 -- by vic-ironport-int.csiro.au with ESMTP; 01 May 2007 15:56:19 +1000 10, 30 -- X-IronPort-AV: i="4.14,472,1170594000"; 10, 30 -- d="scan'208"; a="133087583:sNHT27030780" 10, 30 -- Received: from exvicn1-mel.nexus.csiro.au ([138.194.3.60]) by exgw1-mel.nexus.csiro.au with Microsoft SMTPSVC(5.0.2195.6713); 10, 30 -- Tue, 1 May 2007 15:56:19 +1000 10, 30 -- Received: from exvic5-gex.nexus.csiro.au ([138.194.194.6]) by exvicn1-mel.nexus.csiro.au with Microsoft SMTPSVC(5.0.2195.6713); 10, 30 -- Tue, 1 May 2007 15:56:19 +1000 10, 30 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 10, 30 -- content-class: urn:content-classes:message 10, 30 -- MIME-Version: 1.0 10, 30 -- Content-Type: text/plain; 10, 30 -- charset="us-ascii" 10, 30 -- Subject: imaging plant (carrot) cell walls 10, 30 -- Date: Tue, 1 May 2007 15:56:19 +1000 10, 30 -- Message-ID: {32CDDDAA7161394599F002549491574922B860-at-exvic5-gex.nexus.csiro.au} 10, 30 -- X-MS-Has-Attach: 10, 30 -- X-MS-TNEF-Correlator: 10, 30 -- Thread-Topic: imaging plant (carrot) cell walls 10, 30 -- Thread-Index: AceLtXFS+OmOyyZWTwG1OO/Iv6qY5Q== 10, 30 -- From: {Colin.Veitch-at-csiro.au} 10, 30 -- To: {microscopy-at-msa.microscopy.com} 10, 30 -- X-OriginalArrivalTime: 01 May 2007 05:56:19.0618 (UTC) FILETIME=[7080D020:01C78BB5] 10, 30 -- Content-Transfer-Encoding: 8bit 10, 30 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l415uKdm015200 ==============================End of - Headers==============================
Vibration control is important and our confocal/time-lapse/laser dissection microscopes have at least an in-house made heavy cheap damping plate (140kg) with anti-vibrational pads (which prevent banging on the support worktop wobbling the image on the VDU screen) or at best a professional air-table with the confocals and TIRF. Our facility is on the second floor in busy London and the building has no special anti-vibration features - the opposite really as shutting an outside corridor door can cause a judder on the screen without some sort of isolation system. Stout, well fitted thick worktops with extra support under the microscope, also helps (particularly with 140+kg on it). Very basic lab microscopes (~£5,000) used at 10x to 40x seem to get by with minimal vibration protection, probably as their optics are so poor that is the least of their problems being optimised for under 200x magnification. Our Leica SP2 AOBS came with a natty Leica granite and squash ball number and the PALM micro-dissection system with the heavy plate/rubber pads approach, both similar to our cheap in-house approach, but both work OK up to the full 1,000x mag - i.e. you don't seem to need much more vibrational protection for optical microscopes. This is important as invariably throughout their life the microscopes move slowly about the building from room to room as the wind of departmental politics change.
The main problem we get is people leaning the anti-vibrational tables to write, and things like focus and manual stage drift caused by air temperature variations in the air-conditioned room (although this is minimised by using a large incubator enclosing the stage and objectives for live cell work, and the use of motorised stages), see below.
At UKAEA Harwell we did have a Hitachi low voltage hi-res SEM that was moved, during business 'restructuring', from it's purpose built anti-vibrational room with faraday cage shielding and window blinds, and ended up in the ground floor of our building in a normal lab. The windows were covered with tin foil, and, particularly every time a large vehicle drove by, the image at high mag was unusable (the hi-mag image always wobbled whatever). The old purpose built suite of microscope rooms, at another site, were converted to offices. This was 15 years ago, so I don't know if such SEM's have better vibration protection now, but in this case the very expensive purpose built room was certainly better.
In general vibration is the least of our problems, although all the rooms (even those being built for microscopes) were designed with little thought having tacky thin wobbly worktop fittings. More importantly things like putting the cold feed out vent above the microscope is common to all the rooms and temperature control is only to office standard with variations of typically 4 to 5 oC (and dropping to 16oC overnight with the microscopes off, making the Zeiss/Leica anti-fluorescent immersion go cloudy - heat to 40oC to clear it). A temperature change of 15oC, as the live cell incubator warms up, moves focus out by about 35um, so ambient temperature changes are a very serious problem. This plays havoc with focus and any manual XY stage (both drift). Ideally keep the room fairly warm (22oC, or higher if you use incubators, is ideal) and shield any air-conditioning out vents from the microscope area. In all cases we have had to pay £1,000+ to improve the 'balanced' air condition temperature control system - the art of the Victorian concept of a thermostat, set to 22oC, seems to have been lost. So make sure you get this right. Even a large stage incubator can get into trouble fighting against a poorly setup up air conditioning system. Plus ensure that the exhaust heat from the argon laser of a confocal is suitably ducted out of the building (they give out 3 to 5 kW). Heat generated from these confocals with also test the quality of any room's air conditioning system.
We also get mains interference effects on transmission confocal images with some systems occasionally (according to the manufacturers engineers, but I suspect a dodgey internal connection though). All of our systems have some of rudimentary mains spike/surge filtration and UPS PC protection, and it would be worth thinking of putting quality mains spike protection into the wiring system of a microscope room when it's being built (with UPS being a stand alone box by the PC).
Keith
--------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Institute of Ophthalmology UCL, London EC1V 9EL
-----Original Message----- X-from: ph2-at-sprynet.com [mailto:ph2-at-sprynet.com] Sent: 01 May 2007 03:17 To: keith.morris-at-ucl.ac.uk
1. A recent study was published in Sound and Vibration on Microscope Acceptability of vibration for optical ones. (I'm at home and don't have a copy with me)
- Vibration Sensitivity of Laboratory Bench Microscopes Hal Amick and Matthew Stead, Colin Gordon & Associates
Benchtop optical microscopes are among the most common tools found in R&D facilities. The importance of vibration isolation to maximize image quality is discussed. Vibration criteria are presented.
2. I talked to the author last week (I have always been interested in this since 3D vibration analysis work at GA Tech years ago) about some other things that could be used as criteria for assessment and asked him to submit an abstract for the upcoming Inter-Micro in Chicago, IL in July. Hopefully he'll make it. In the mean time if you need a copy of the article contact me or the S&V journal (It's not on their website yet http://www.sandv.com/home.htm ).
3. This article does not cover the building aspects, but a good vibration engineer could help there in conjunction with a structural engineer, particularly in combination with the frequency response data from this article.
Tony
..................................................................... Andrew Anthony "Tony" Havics, CHMM, CIH, PE pH2, LLC 5250 E US 36, Suite 830 Avon, IN 46123 (317) 718-7020 off (317) 718-7038 fax (317) 409-3238 cell
90% of Risk Management is knowing where to place the decimal point...any consultant can give you the other 10%(SM)
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-----Original Message----- X-from: sandra.filippi-at-montgomerycollege.edu [mailto:sandra.filippi-at-montgomerycollege.edu] Sent: Monday, April 30, 2007 9:21 PM To: ph2-at-sprynet.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sandra.filippi-at-montgomerycollege.edu) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, April 30, 2007 at 20:12:50 ---------------------------------------------------------------------------
Email: sandra.filippi-at-montgomerycollege.edu Name: Sandra Lee Filippi, Campus Planner
Organization: montgomery college
Education: Graduate College
Location: Rockville MD
Question:
Dear Sir or Madam Microscopist:
Montgomery College is in design development for a new academic teaching science center. The lab planner for the design team is citing ASHRAE TC2.6 Guideline Criteria for use in Planning New Facilities and insists that we design to the VC-B level because the biology department uses compound microscopes with total magnification of 1,000x (oil immersion). I managed academic science labs for a two-year college for more than 20 years. We routinely used light microscopes at total magnification of 1,000x (oil immersion) in a facility built in 1965 without special vibration control. When we opened our new science center in 1995 it did not have special vibration control either. I have studied at Georgetown University and did student research at The NIH. Neither of these facilities required special vibration control for a light microscope. I have consulted with the Westover Scientific Customer Service/Product Specialist ñ Microscopy who states, ìTo be honest with you I have never heard of such a thing, neither have my colleagues. ÝThere are tables that are manufactured to reduce vibrations, but Iíve never heard of a building being designed around a microscope. ÝWe have customers who use microscopes in old buildings all over the world. Westover recommends that a microscope be used on a vibration free surface which typically means not locating your microscope on a table with other equipment that can cause vibrations such as fans and other lab equipment. Microscopes have been used for years and years on standard benchtops and other tables without issue. It's not our position to recommend additional support when building a new structure. In my entire career I have never heard of this requirement.î Westover manufactures microscopes for Fisher Scientific. The lab planner is going to cost us dearly unless I can refute his recommendation. What is your professional opinion in this matter? Thank you very much for your time. Sandra
Sandra Lee Filippi, Campus Planner Montgomery College Suite 200 40 West Gude Drive Room 223 Rockville MD 20850-1166 301.251.7362 vox 301.251.7379 fax 240.882.6672 cell
May I please have a copy of the article? I am able to receive scanned (PDF) attachments to email and/or fax to the numbers in my signature. THANK YOU.
Sandra
Sandra Lee Filippi, Campus Planner
Suite 200 40 West Gude Drive Room 223 Rockville MD 20850-1166 301.251.7362 vox 301.251.7379 fax 240.882.6672 cell sandra.filippi-at-montgomerycollege.edu
-----Original Message----- X-from: Tony Havics [mailto:ph2-at-sprynet.com] Sent: Monday, April 30, 2007 10:13 PM To: Filippi, Sandra; Micrscopy Listserve
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sandra.filippi-at-montgomerycollege.edu) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, April 30, 2007 at 20:12:50 ---------------------------------------------------------------------------
Email: sandra.filippi-at-montgomerycollege.edu Name: Sandra Lee Filippi, Campus Planner
Organization: montgomery college
Education: Graduate College
Location: Rockville MD
Question:
Dear Sir or Madam Microscopist:
Montgomery College is in design development for a new academic teaching science center. The lab planner for the design team is citing ASHRAE TC2.6 Guideline Criteria for use in Planning New Facilities and insists that we design to the VC-B level because the biology department uses compound microscopes with total magnification of 1,000x (oil immersion). I managed academic science labs for a two-year college for more than 20 years. We routinely used light microscopes at total magnification of 1,000x (oil immersion) in a facility built in 1965 without special vibration control. When we opened our new science center in 1995 it did not have special vibration control either. I have studied at Georgetown University and did student research at The NIH. Neither of these facilities required special vibration control for a light microscope. I have consulted with the Westover Scientific Customer Service/Product Specialist ñ Microscopy who states, ìTo be honest with you I have never heard of such a thing, neither have my colleagues. ÝThere are tables that are manufactured to reduce vibrations, but Iíve never heard of a building being designed around a microscope. ÝWe have customers who use microscopes in old buildings all over the world. Westover recommends that a microscope be used on a vibration free surface which typically means not locating your microscope on a table with other equipment that can cause vibrations such as fans and other lab equipment. Microscopes have been used for years and years on standard benchtops and other tables without issue. It's not our position to recommend additional support when building a new structure. In my entire career I have never heard of this requirement.î Westover manufactures microscopes for Fisher Scientific. The lab planner is going to cost us dearly unless I can refute his recommendation. What is your professional opinion in this matter? Thank you very much for your time. Sandra
Sandra Lee Filippi, Campus Planner Montgomery College Suite 200 40 West Gude Drive Room 223 Rockville MD 20850-1166 301.251.7362 vox 301.251.7379 fax 240.882.6672 cell
G'day Colin, I will assume that when you wrote "some carrots" you mean carrot roots, the big bright orange things we eat. I can give you a couple of suggestions although clearly without knowing more about the goals I am a little in the dark.
1. Good old polarized light microscopy. As I am sure you know from fibers (i.e., fibres), cellulose is birefringent. If you embed the roots in your favorite resin, cut semi thin sections (don't stain), you can then image in polarized light. This can give you net orientation (mean extinction angle) and information about the extent of orientation (retardance). Note that while mature xylem fibers/vessels in the carrot root will have a retardance on the order of cotton, that of the rank and file cells in the root will be much less, so you need a reasonably sensitive polarized light set up.
2. SEM. A nice trick to image the inner most layer of the cell wall with SEM is to make fresh sections. By this I mean cut the roots before you fix them. The turgor pressure ejects most/all of the cell contents, including the plasma membrane. Cut the sections in somthing simple like PBS and rinse for 10 min. I have never tried this with carrot roots but on a variety of other plant tissues, this works. What you do next depends on your instrument. In my lab, we fix, dehydrate in ethanol, do CPD, coat with Pt, and image with high vac FESEM. This shows cell wall structure nicely. Microfibrillar structures revealed in this way are on on order of 10 nm. There has been no extraction so they contain more than cellulose. With a VP instrument, you might be able to skip all that and image directly, right after rinsing. You may or may not be able to image these reliably in VP mode.
Hope this helps. Feel free to contact me with questions.
Tobias
} } } G'day, } } We have been asked to look at the cell wall structure of some carrots } and have essentially no experience in this area at all. At this stage } we are interested in the cellulose structure to start with. } } We have access to a Hitachi VP FESEM and JEOL TEM as well as a range of } optical techniques. What would be the best technique for sample } preparation (CPD etc) and imaging? } } Thank you very much in advance for your help with this. } } Colin Veitch } } Electron Microscopist } CSIRO Textile and Fibre Technology } PO Box 21, BELMONT, Vic. 3216. Australia. } colin.veitch-at-csiro.au } http://www.tft.csiro.au } } +61 (0) 3 5246 4000 } 0438 538 475 } +61 (0) 3 5246 4811 } } The information contained in this e-mail message may be privileged or } confidential information. If you are not an intended recipient, you may } not copy, distribute or take any action in reliance on it. If you have } received this message in error, please telephone CSIRO Textile and Fibre } Technology on +61 3 5246 4000. } } } } ==============================Original Headers============================== } 10, 30 -- From Colin.Veitch-at-csiro.au Tue May 1 00:56:22 2007 } 10, 30 -- Received: from vic-MTAout3.csiro.au (vic-MTAout3.csiro.au } [150.229.64.39]) } 10, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id l415uKdm015200 } 10, 30 -- for {microscopy-at-msa.microscopy.com} ; Tue, 1 May 2007 } 00:56:21 -0500 } 10, 30 -- DomainKey-Signature: s=email; d=csiro.au; c=nofws; q=dns; } b=uZBrvaD0cxMV9MbIOdyLEoSa9e/31sIKDfJqM+mVQ4PGmU2m4Cex00xvVXHhJrzY5SufCwEgczAoIx1gjnxnvfVGJ+Vtlh8poqgarafPm4Vk8DsxYJn4bPGmCcNy6pjO; } 10, 30 -- Received: from exgw1-mel.nexus.csiro.au ([138.194.3.56]) } 10, 30 -- by vic-ironport-int.csiro.au with ESMTP; 01 May 2007 } 15:56:19 +1000 } 10, 30 -- X-IronPort-AV: i="4.14,472,1170594000"; } 10, 30 -- d="scan'208"; a="133087583:sNHT27030780" } 10, 30 -- Received: from exvicn1-mel.nexus.csiro.au ([138.194.3.60]) } by exgw1-mel.nexus.csiro.au with Microsoft SMTPSVC(5.0.2195.6713); } 10, 30 -- Tue, 1 May 2007 15:56:19 +1000 } 10, 30 -- Received: from exvic5-gex.nexus.csiro.au ([138.194.194.6]) } by exvicn1-mel.nexus.csiro.au with Microsoft SMTPSVC(5.0.2195.6713); } 10, 30 -- Tue, 1 May 2007 15:56:19 +1000 } 10, 30 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 } 10, 30 -- content-class: urn:content-classes:message } 10, 30 -- MIME-Version: 1.0 } 10, 30 -- Content-Type: text/plain; } 10, 30 -- charset="us-ascii" } 10, 30 -- Subject: imaging plant (carrot) cell walls } 10, 30 -- Date: Tue, 1 May 2007 15:56:19 +1000 } 10, 30 -- Message-ID: } {32CDDDAA7161394599F002549491574922B860-at-exvic5-gex.nexus.csiro.au} } 10, 30 -- X-MS-Has-Attach: } 10, 30 -- X-MS-TNEF-Correlator: } 10, 30 -- Thread-Topic: imaging plant (carrot) cell walls } 10, 30 -- Thread-Index: AceLtXFS+OmOyyZWTwG1OO/Iv6qY5Q== } 10, 30 -- From: {Colin.Veitch-at-csiro.au} } 10, 30 -- To: {microscopy-at-msa.microscopy.com} } 10, 30 -- X-OriginalArrivalTime: 01 May 2007 05:56:19.0618 (UTC) } FILETIME=[7080D020:01C78BB5] } 10, 30 -- Content-Transfer-Encoding: 8bit } 10, 30 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } ns.microscopy.com id l415uKdm015200 } ==============================End of - Headers==============================
I fear that Desktop Microscopist is no more. I have tried, with no luck, to contact the source; Lacuna Labs. The authors did not respond to my email, after I searched them out, several months ago, when the computer my application ran on died.
I have used that particular application for years in my work with Laue x-ray backscatter diffraction. The version I have is 2.1.x from 1998. It will not work on an operating system beyond 9.x and my onsite computer geeks don't support that old stuff anymore.
I now use another application called ScanOrient to back up my DM information. But it is not quite the same. --
+++++++++++++++++++++++++++++
R. Ann Bliss Technologist, Chemistry Materials and Life Science Materials Science and Technology Division Lawrence Livermore National Laboratory
_____________________________
==============================Original Headers============================== 7, 19 -- From bliss5-at-llnl.gov Tue May 1 11:42:04 2007 7, 19 -- Received: from nspiron-1.llnl.gov (nspiron-1.llnl.gov [128.115.41.81]) 7, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l41Gg46L007371 7, 19 -- for {microscopy-at-microscopy.com} ; Tue, 1 May 2007 11:42:04 -0500 7, 19 -- Received: from altaite.llnl.gov (HELO [134.9.108.10]) ([134.9.108.10]) 7, 19 -- by nspiron-1.llnl.gov with ESMTP; 01 May 2007 09:42:02 -0700 7, 19 -- X-Attachments: 7, 19 -- X-IronPort-AV: i="4.14,475,1170662400"; 7, 19 -- d="scan'208"; a="24940328:sNHT26479220" 7, 19 -- Mime-Version: 1.0 7, 19 -- Message-Id: {p06230906c25d1b9b7c46-at-[134.9.108.10]} 7, 19 -- In-Reply-To: {200704271242.l3RCg1LD014448-at-ns.microscopy.com} 7, 19 -- References: {200704271242.l3RCg1LD014448-at-ns.microscopy.com} 7, 19 -- Date: Tue, 1 May 2007 09:42:00 -0700 7, 19 -- To: USTweety-at-hotmail.com 7, 19 -- From: "R. Ann Bliss" {bliss5-at-llnl.gov} 7, 19 -- Subject: Re: [Microscopy] viaWWW: desktop microscopist 7, 19 -- Cc: microscopy-at-microscopy.com 7, 19 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
Anyone out there have any advice on how to enhance the contrast and definition of the membranes of the cristae of mitochondria? The samples brought to me are monolayer cell cultures of cancer cells grown on Thermanox coverslips. This is how I'm currently processing the samples: Primary fixation is 2% glutaraldehyde, 2% paraformaldehyde, 0.5% acrolein in 0.1M Sorensen's phosphate buffer pH 7.2. Post-fixation in 1%Osmium Tetroxide in 0.1M Sorensen's phosphate buffer. Dehydration in 50, 70, 90, 95, 100%X3 ethanol solutions. embedding in Araldite. Sections are stained with 2% uranyl acetate aqueous 15 minutes and Reynold's lead citrate 10 minutes. The density of the cytoplasm and mitochondrial matrix are similar with the result that the contrast of the mitochondria is similar to the cytoplasm. The mitochondria and it's membranes (outer and that of the cristae) don't really "stand out". The researchers involved want to see really contrasty mitochondrial cristae.
The next thing I'm going to try is post-fixation with a mix of osmium tetroxide and potassium ferrocyanide.
Anyone have any other ideas? Thanks in advance.
Tom Bargar University of Nebraska Medical Center Core Electron Microscopy Research Facility 986395 Nebraska Medical Center Omaha, NE 68198-6395 402-559-7347 tbargar-at-unmc.edu
==============================Original Headers============================== 8, 20 -- From tbargar-at-unmc.edu Tue May 1 15:39:39 2007 8, 20 -- Received: from zixvpm02.unmc.edu (zixvpm02.unmc.edu [192.198.54.127]) 8, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l41KddQH023357 8, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 1 May 2007 15:39:39 -0500 8, 20 -- Received: from zixvpm02.unmc.edu (ZixVPM [127.0.0.1]) 8, 20 -- by Outbound.unmc.edu (Proprietary) with ESMTP id 8A304A0061 8, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 1 May 2007 15:39:39 -0500 (CDT) 8, 20 -- Received: from unmcnotes01.unmc.edu (host-137-197-103-35.unmc.edu [137.197.103.35]) 8, 20 -- by zixvpm02.unmc.edu (Proprietary) with ESMTP id 72E1039804A 8, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 1 May 2007 15:39:38 -0500 (CDT) 8, 20 -- Subject: Help with enhancing contrast of Mitochondrial cristae 8, 20 -- To: Microscopy-at-MSA.Microscopy.Com 8, 20 -- X-Mailer: Lotus Notes Release 7.0.1 January 17, 2006 8, 20 -- Message-ID: {OFDC8670F1.FEB83264-ON862572CE.006F5771-862572CE.00717D6E-at-unmc.edu} 8, 20 -- From: Tom W Bargar {tbargar-at-unmc.edu} 8, 20 -- Date: Tue, 1 May 2007 15:39:27 -0500 8, 20 -- X-MIMETrack: Serialize by Router on UNMCNOTES01.UNMC.EDU/Servers/UNEBR at 05/01/2007 03:39:38 8, 20 -- PM 8, 20 -- MIME-Version: 1.0 8, 20 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
Dear Tom- You anticipated one of my responses by stating that you are going to use K-Ferrocyanide with the osmium. Another thing you can try (along with the Os/K-fer) is to use the following as your primary fix: 4% pfa, 2.5% glut, 0.002% picric acid in No-Cacod. buffer (original ref: It & Karnovsky J Cell Biol Vol 89 Abstract #418, 1968).
I was first told about this fix by a group who studied outer rod segments of the eye....LOTS of membranes!
Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Weill-Cornell Medical College
I need to stain nucleic acids in the 540 nm excitation range on semi thin cryosections. I selected SYTO 83 from Molecular Probes(exc. 543, emission 559), but so far was not very successful - the cytoplasm actually shows more staining than the nucleus. They are not very specific about the protocol (except that one should not use phosphate buffers). Does anybody there in cyberspace have any experience with these dyes?
Thanks,
-- Michal Jarnik
==============================Original Headers============================== 4, 17 -- From Michal.Jarnik-at-fccc.edu Tue May 1 16:31:26 2007 4, 17 -- Received: from azah.fccc.edu (azah.fccc.edu [131.249.4.237]) 4, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l41LVQpe014416 4, 17 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 1 May 2007 16:31:26 -0500 4, 17 -- Received: from [10.40.12.93] (emf1.dyn.fccc.edu [10.40.12.93]) 4, 17 -- (authenticated bits=0) 4, 17 -- by azah.fccc.edu (8.13.6/8.13.6) with ESMTP id l41LVP2l014359 4, 17 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 1 May 2007 17:31:25 -0400 (EDT) 4, 17 -- Message-ID: {4637B1AB.2020905-at-fccc.edu} 4, 17 -- Date: Tue, 01 May 2007 17:31:23 -0400 4, 17 -- From: Michal Jarnik {Michal.Jarnik-at-fccc.edu} 4, 17 -- User-Agent: Thunderbird 1.5.0.5 (Macintosh/20060719) 4, 17 -- MIME-Version: 1.0 4, 17 -- To: Microscopy-at-MSA.Microscopy.Com 4, 17 -- Subject: SYTO Orange stains 4, 17 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 17 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Mix in the following proportions: 0.2g UA 3ml 50% ethanol
Let mix at least several hours on rocker/shaker. Next day, try a series of UA staining times and examine (say, every 5 minutes). I've found 10-15 minutes optimal, as longer times appeared to also darken surrounding plastic matrix. Pick your optimal time, stain, rinse, and counterstain with Reynold's LC, ~5-10 minutes, rinse.
Post-fixing in reduced osmium as you state is my other suggestion, but see if ethanolic UA gives you what you need before re-embedding more samples. Also see if the two techniques combined (reduced osmium, ethanolic UA stain) give you even better results!
Be sure to post your findings. Best regards,
Walter F. Bobrowski Sr. Scientist Pfizer Global R&D 2800 Plymouth Rd. Ann Arbor, MI 48105
TEL: 734-622-7814 FAX: 734-622-5718
"Like the strength of a steel rod, the true character of a person can only be known under extreme stress." - Leslie Fieger
-----Original Message----- X-from: tbargar-at-unmc.edu [mailto:tbargar-at-unmc.edu] Sent: Tuesday, May 01, 2007 4:46 PM To: Bobrowski, Walter
Dear listers,
Anyone out there have any advice on how to enhance the contrast and definition of the membranes of the cristae of mitochondria? The samples brought to me are monolayer cell cultures of cancer cells grown on Thermanox coverslips. This is how I'm currently processing the samples: Primary fixation is 2% glutaraldehyde, 2% paraformaldehyde, 0.5% acrolein in 0.1M Sorensen's phosphate buffer pH 7.2. Post-fixation in 1%Osmium Tetroxide in 0.1M Sorensen's phosphate buffer. Dehydration in 50, 70, 90, 95, 100%X3 ethanol solutions. embedding in Araldite. Sections are stained with 2% uranyl acetate aqueous 15 minutes and Reynold's lead citrate 10 minutes. The density of the cytoplasm and mitochondrial matrix are similar with the result that the contrast of the mitochondria is similar to the cytoplasm. The mitochondria and it's membranes (outer and that of the cristae) don't really "stand out". The researchers involved want to see really contrasty mitochondrial cristae.
The next thing I'm going to try is post-fixation with a mix of osmium tetroxide and potassium ferrocyanide.
Anyone have any other ideas? Thanks in advance.
Tom Bargar University of Nebraska Medical Center Core Electron Microscopy Research Facility 986395 Nebraska Medical Center Omaha, NE 68198-6395 402-559-7347 tbargar-at-unmc.edu
==============================Original Headers============================== 8, 20 -- From tbargar-at-unmc.edu Tue May 1 15:39:39 2007 8, 20 -- Received: from zixvpm02.unmc.edu (zixvpm02.unmc.edu [192.198.54.127]) 8, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l41KddQH023357 8, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 1 May 2007 15:39:39 -0500 8, 20 -- Received: from zixvpm02.unmc.edu (ZixVPM [127.0.0.1]) 8, 20 -- by Outbound.unmc.edu (Proprietary) with ESMTP id 8A304A0061 8, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 1 May 2007 15:39:39 -0500 (CDT) 8, 20 -- Received: from unmcnotes01.unmc.edu (host-137-197-103-35.unmc.edu [137.197.103.35]) 8, 20 -- by zixvpm02.unmc.edu (Proprietary) with ESMTP id 72E1039804A 8, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 1 May 2007 15:39:38 -0500 (CDT) 8, 20 -- Subject: Help with enhancing contrast of Mitochondrial cristae 8, 20 -- To: Microscopy-at-MSA.Microscopy.Com 8, 20 -- X-Mailer: Lotus Notes Release 7.0.1 January 17, 2006 8, 20 -- Message-ID: {OFDC8670F1.FEB83264-ON862572CE.006F5771-862572CE.00717D6E-at-unmc.edu} 8, 20 -- From: Tom W Bargar {tbargar-at-unmc.edu} 8, 20 -- Date: Tue, 1 May 2007 15:39:27 -0500 8, 20 -- X-MIMETrack: Serialize by Router on UNMCNOTES01.UNMC.EDU/Servers/UNEBR at 05/01/2007 03:39:38 8, 20 -- PM 8, 20 -- MIME-Version: 1.0 8, 20 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
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==============================Original Headers============================== 25, 31 -- From Walter.Bobrowski-at-pfizer.com Wed May 2 07:33:14 2007 25, 31 -- Received: from mopmsgoa02.pfizer.com (mopmsgo.pfizer.com [148.168.100.84]) 25, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l42CXDfg007274 25, 31 -- for {microscopy-at-microscopy.com} ; Wed, 2 May 2007 07:33:14 -0500 25, 31 -- Received: from groamrexc02.amer.pfizer.com (groamrexc02.amer.pfizer.com [172.30.8.169]) 25, 31 -- by mopmsgoa02.pfizer.com (8.13.7/8.13.7) with ESMTP id l42CXDKK012936; 25, 31 -- Wed, 2 May 2007 08:33:13 -0400 25, 31 -- Received: from mopamrexc02.amer.pfizer.com ([170.116.30.68]) by groamrexc02.amer.pfizer.com with Microsoft SMTPSVC(6.0.3790.1830); 25, 31 -- Wed, 2 May 2007 08:33:13 -0400 25, 31 -- Received: from anaamrexm01.amer.pfizer.com ([162.48.101.10]) by mopamrexc02.amer.pfizer.com with Microsoft SMTPSVC(6.0.3790.1830); 25, 31 -- Wed, 2 May 2007 08:33:12 -0400 25, 31 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 25, 31 -- Content-class: urn:content-classes:message 25, 31 -- MIME-Version: 1.0 25, 31 -- Content-Type: text/plain; 25, 31 -- charset="us-ascii" 25, 31 -- Subject: RE: [Microscopy] Help with enhancing contrast of Mitochondrial cristae 25, 31 -- Date: Wed, 2 May 2007 08:33:05 -0400 25, 31 -- Message-ID: {CF89AF6ABD93B746A55BCCD9480A4B3D0909BB7D-at-anaamrexm01.amer.pfizer.com} 25, 31 -- In-Reply-To: {200705012046.l41KkOL4032172-at-ns.microscopy.com} 25, 31 -- X-MS-Has-Attach: 25, 31 -- X-MS-TNEF-Correlator: 25, 31 -- Thread-Topic: [Microscopy] Help with enhancing contrast of Mitochondrial cristae 25, 31 -- thread-index: AceMMckRQ+3zgA8uRPOzn5TbyIEhgwAgrR8g 25, 31 -- From: "Bobrowski, Walter" {Walter.Bobrowski-at-pfizer.com} 25, 31 -- To: {tbargar-at-unmc.edu} , {microscopy-at-microscopy.com} 25, 31 -- X-OriginalArrivalTime: 02 May 2007 12:33:12.0921 (UTC) FILETIME=[0CC03C90:01C78CB6] 25, 31 -- X-Proofpoint-Virus-Version: vendor=fsecure engine=4.65.5502:2.3.11,1.2.37,4.0.164 definitions=2007-05-02_03:2007-04-28,2007-05-02,2007-05-02 signatures=0 25, 31 -- X-Proofpoint-Spam-Reason: safe 25, 31 -- Content-Transfer-Encoding: 8bit 25, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l42CXDfg007274 ==============================End of - Headers==============================
It is possible that you are losing your cellular membranes during the dehydration steps. First, en bloc stain with saturated uranyl acetate in water and then go through the dehydration steps quickly, about 2 minutes for each step. Begin with 50 % EtOH and end with only one change of 100 % EtOH. If you are infiltrating with EtOH: araldite, keep the steps with high EtOH content short, too, about a 30 minute maximum.
I hope that helps!
Dotty Sorenson
On May 1, 2007, at 4:44 PM, tbargar-at-unmc.edu wrote:
} } } } ---------------------------------------------------------------------- } ------ } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } } Dear listers, } } Anyone out there have any advice on how to enhance the contrast and } definition of the membranes of the cristae of mitochondria? The } samples } brought to me are monolayer cell cultures of cancer cells grown on } Thermanox coverslips. This is how I'm currently processing the } samples: } Primary fixation is 2% glutaraldehyde, 2% paraformaldehyde, 0.5% } acrolein } in 0.1M Sorensen's phosphate buffer pH 7.2. Post-fixation in 1%Osmium } Tetroxide in 0.1M Sorensen's phosphate buffer. Dehydration in 50, } 70, 90, } 95, 100%X3 ethanol solutions. embedding in Araldite. Sections are } stained } with 2% uranyl acetate aqueous 15 minutes and Reynold's lead } citrate 10 } minutes. The density of the cytoplasm and mitochondrial matrix are } similar with the result that the contrast of the mitochondria is } similar to } the cytoplasm. The mitochondria and it's membranes (outer and that } of the } cristae) don't really "stand out". The researchers involved want } to see } really contrasty mitochondrial cristae. } } The next thing I'm going to try is post-fixation with a mix of osmium } tetroxide and potassium ferrocyanide. } } Anyone have any other ideas? Thanks in advance. } } } Tom Bargar } University of Nebraska Medical Center } Core Electron Microscopy Research Facility } 986395 Nebraska Medical Center } Omaha, NE 68198-6395 } 402-559-7347 } tbargar-at-unmc.edu } } } ==============================Original } Headers============================== } 8, 20 -- From tbargar-at-unmc.edu Tue May 1 15:39:39 2007 } 8, 20 -- Received: from zixvpm02.unmc.edu (zixvpm02.unmc.edu } [192.198.54.127]) } 8, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP } id l41KddQH023357 } 8, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 1 May 2007 } 15:39:39 -0500 } 8, 20 -- Received: from zixvpm02.unmc.edu (ZixVPM [127.0.0.1]) } 8, 20 -- by Outbound.unmc.edu (Proprietary) with ESMTP id 8A304A0061 } 8, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 1 May 2007 } 15:39:39 -0500 (CDT) } 8, 20 -- Received: from unmcnotes01.unmc.edu } (host-137-197-103-35.unmc.edu [137.197.103.35]) } 8, 20 -- by zixvpm02.unmc.edu (Proprietary) with ESMTP id 72E1039804A } 8, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 1 May 2007 } 15:39:38 -0500 (CDT) } 8, 20 -- Subject: Help with enhancing contrast of Mitochondrial } cristae } 8, 20 -- To: Microscopy-at-MSA.Microscopy.Com } 8, 20 -- X-Mailer: Lotus Notes Release 7.0.1 January 17, 2006 } 8, 20 -- Message-ID: {OFDC8670F1.FEB83264-ON862572CE. } 006F5771-862572CE.00717D6E-at-unmc.edu} } 8, 20 -- From: Tom W Bargar {tbargar-at-unmc.edu} } 8, 20 -- Date: Tue, 1 May 2007 15:39:27 -0500 } 8, 20 -- X-MIMETrack: Serialize by Router on UNMCNOTES01.UNMC.EDU/ } Servers/UNEBR at 05/01/2007 03:39:38 } 8, 20 -- PM } 8, 20 -- MIME-Version: 1.0 } 8, 20 -- Content-type: text/plain; charset=US-ASCII } ==============================End of - } Headers============================== } }
Dorothy Sorenson Microscopy and Image-analysis Laboratory Department of Cell and Developmental Biology University Of Michigan Medical School A807 BSRB 109 Zina Pitcher Place Ann Arbor, MI 48109-2200 (734)763-1170 FAX (734)763-1166
==============================Original Headers============================== 9, 19 -- From dsoren-at-umich.edu Wed May 2 10:52:50 2007 9, 19 -- Received: from hellskitchen.mr.itd.umich.edu (smtp.mail.umich.edu [141.211.14.82]) 9, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l42FqofY022545 9, 19 -- for {microscopy-at-msa.microscopy.com} ; Wed, 2 May 2007 10:52:50 -0500 9, 19 -- Received: FROM [10.21.129.180] (host-18.subnet-17.med.umich.edu [141.214.17.18]) 9, 19 -- BY hellskitchen.mr.itd.umich.edu ID 4638B395.32DC9.16753 ; 9, 19 -- 2 May 2007 11:51:49 -0400 9, 19 -- In-Reply-To: {200705012044.l41KibJi029434-at-ns.microscopy.com} 9, 19 -- References: {200705012044.l41KibJi029434-at-ns.microscopy.com} 9, 19 -- Mime-Version: 1.0 (Apple Message framework v752.2) 9, 19 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 9, 19 -- Message-Id: {F26C58BE-25AD-4A13-960D-B8BEAB3420E6-at-umich.edu} 9, 19 -- Cc: microscopy-at-msa.microscopy.com 9, 19 -- Content-Transfer-Encoding: 7bit 9, 19 -- From: Dorothy Sorenson {dsoren-at-umich.edu} 9, 19 -- Subject: Re: [Microscopy] Help with enhancing contrast of Mitochondrial cristae 9, 19 -- Date: Wed, 2 May 2007 11:48:33 -0400 9, 19 -- To: tbargar-at-unmc.edu 9, 19 -- X-Mailer: Apple Mail (2.752.2) ==============================End of - Headers==============================
Along this same line of thought, if you have access to a variable-wattage laboratory microwave suitable for histo- or EM processing, you can drastically shorten your dehydration solvent exposure times to seconds, rather than half an hour or more.
Processing times for all steps are greatly reduced by using microwaves. It is possible to go from fresh sample to polymerized blocks in 4-5 hours, or sometimes less, with often superior results compared to conventional processing. Extraction of sample components is minimized by the short exposures to the various reagents, especially in dehydration steps.
You can find our "generic" microwave protocol at http://www.emc.missouri.edu/Pdfs/General%202-ME%20Microwave%20Processing %20Protocol.pdf. Like most everything else in EM work, it gets modified all the time, but it's a good starting point.
Good luck, Randy
Randy Tindall Senior EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount= Week&NavType=Both&Type=TimePlan
-----Original Message----- X-from: dsoren-at-umich.edu [mailto:dsoren-at-umich.edu] Sent: Wednesday, May 02, 2007 10:55 AM To: Tindall, Randy D.
Dear Tom,
It is possible that you are losing your cellular membranes during the dehydration steps. First, en bloc stain with saturated uranyl acetate in water and then go through the dehydration steps quickly, about 2 minutes for each step. Begin with 50 % EtOH and end with only one change of 100 % EtOH. If you are infiltrating with EtOH: araldite, keep the steps with high EtOH content short, too, about a 30 minute maximum.
I hope that helps!
Dotty Sorenson
On May 1, 2007, at 4:44 PM, tbargar-at-unmc.edu wrote:
} } } } ---------------------------------------------------------------------- } ------ } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- http://www.microscopy.com/ } MicroscopyListserver On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } } Dear listers, } } Anyone out there have any advice on how to enhance the contrast and } definition of the membranes of the cristae of mitochondria? The } samples brought to me are monolayer cell cultures of cancer cells } grown on Thermanox coverslips. This is how I'm currently processing } the } samples: } Primary fixation is 2% glutaraldehyde, 2% paraformaldehyde, 0.5% } acrolein in 0.1M Sorensen's phosphate buffer pH 7.2. Post-fixation in
} 1%Osmium Tetroxide in 0.1M Sorensen's phosphate buffer. Dehydration } in 50, 70, 90, 95, 100%X3 ethanol solutions. embedding in Araldite. } Sections are stained with 2% uranyl acetate aqueous 15 minutes and } Reynold's lead citrate 10 } minutes. The density of the cytoplasm and mitochondrial matrix are } similar with the result that the contrast of the mitochondria is } similar to the cytoplasm. The mitochondria and it's membranes (outer } and that of the } cristae) don't really "stand out". The researchers involved want to } see really contrasty mitochondrial cristae. } } The next thing I'm going to try is post-fixation with a mix of osmium } tetroxide and potassium ferrocyanide. } } Anyone have any other ideas? Thanks in advance. } } } Tom Bargar } University of Nebraska Medical Center } Core Electron Microscopy Research Facility } 986395 Nebraska Medical Center } Omaha, NE 68198-6395 } 402-559-7347 } tbargar-at-unmc.edu } } } ==============================Original } Headers============================== } 8, 20 -- From tbargar-at-unmc.edu Tue May 1 15:39:39 2007 8, 20 -- } Received: from zixvpm02.unmc.edu (zixvpm02.unmc.edu } [192.198.54.127]) } 8, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP } id l41KddQH023357 } 8, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 1 May 2007 } 15:39:39 -0500 } 8, 20 -- Received: from zixvpm02.unmc.edu (ZixVPM [127.0.0.1]) } 8, 20 -- by Outbound.unmc.edu (Proprietary) with ESMTP id 8A304A0061 } 8, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 1 May 2007 } 15:39:39 -0500 (CDT) } 8, 20 -- Received: from unmcnotes01.unmc.edu } (host-137-197-103-35.unmc.edu [137.197.103.35]) } 8, 20 -- by zixvpm02.unmc.edu (Proprietary) with ESMTP id 72E1039804A } 8, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 1 May 2007 } 15:39:38 -0500 (CDT) } 8, 20 -- Subject: Help with enhancing contrast of Mitochondrial } cristae 8, 20 -- To: Microscopy-at-MSA.Microscopy.Com 8, 20 -- X-Mailer: } Lotus Notes Release 7.0.1 January 17, 2006 8, 20 -- Message-ID: } {OFDC8670F1.FEB83264-ON862572CE. } 006F5771-862572CE.00717D6E-at-unmc.edu} } 8, 20 -- From: Tom W Bargar {tbargar-at-unmc.edu} 8, 20 -- Date: Tue, 1 } May 2007 15:39:27 -0500 8, 20 -- X-MIMETrack: Serialize by Router on } UNMCNOTES01.UNMC.EDU/ Servers/UNEBR at 05/01/2007 03:39:38 8, 20 -- } PM 8, 20 -- MIME-Version: 1.0 8, 20 -- Content-type: text/plain; } charset=US-ASCII ==============================End of - } Headers============================== } }
Dorothy Sorenson Microscopy and Image-analysis Laboratory Department of Cell and Developmental Biology University Of Michigan Medical School A807 BSRB 109 Zina Pitcher Place Ann Arbor, MI 48109-2200 (734)763-1170 FAX (734)763-1166
==============================Original Headers============================== 9, 19 -- From dsoren-at-umich.edu Wed May 2 10:52:50 2007 9, 19 -- Received: from hellskitchen.mr.itd.umich.edu (smtp.mail.umich.edu [141.211.14.82]) 9, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l42FqofY022545 9, 19 -- for {microscopy-at-msa.microscopy.com} ; Wed, 2 May 2007 10:52:50 -0500 9, 19 -- Received: FROM [10.21.129.180] (host-18.subnet-17.med.umich.edu [141.214.17.18]) 9, 19 -- BY hellskitchen.mr.itd.umich.edu ID 4638B395.32DC9.16753 ; 9, 19 -- 2 May 2007 11:51:49 -0400 9, 19 -- In-Reply-To: {200705012044.l41KibJi029434-at-ns.microscopy.com} 9, 19 -- References: {200705012044.l41KibJi029434-at-ns.microscopy.com} 9, 19 -- Mime-Version: 1.0 (Apple Message framework v752.2) 9, 19 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 9, 19 -- Message-Id: {F26C58BE-25AD-4A13-960D-B8BEAB3420E6-at-umich.edu} 9, 19 -- Cc: microscopy-at-msa.microscopy.com 9, 19 -- Content-Transfer-Encoding: 7bit 9, 19 -- From: Dorothy Sorenson {dsoren-at-umich.edu} 9, 19 -- Subject: Re: [Microscopy] Help with enhancing contrast of Mitochondrial cristae 9, 19 -- Date: Wed, 2 May 2007 11:48:33 -0400 9, 19 -- To: tbargar-at-unmc.edu 9, 19 -- X-Mailer: Apple Mail (2.752.2) ==============================End of - Headers==============================
==============================Original Headers============================== 21, 25 -- From TindallR-at-missouri.edu Wed May 2 11:46:18 2007 21, 25 -- Received: from um-tsmtpout1.um.umsystem.edu (um-tsmtpout1.um.umsystem.edu [209.106.228.23]) 21, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l42GkHUi002675 21, 25 -- for {microscopy-at-microscopy.com} ; Wed, 2 May 2007 11:46:18 -0500 21, 25 -- Received: from UM-XMAIL08.um.umsystem.edu ([209.106.228.34]) by um-tsmtpout1.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 21, 25 -- Wed, 2 May 2007 11:46:17 -0500 21, 25 -- x-mimeole: Produced By Microsoft Exchange V6.5 21, 25 -- Content-class: urn:content-classes:message 21, 25 -- MIME-Version: 1.0 21, 25 -- Content-Type: text/plain; 21, 25 -- charset="us-ascii" 21, 25 -- Subject: RE: [Microscopy] Re: Help with enhancing contrast of Mitochondrial cristae 21, 25 -- Date: Wed, 2 May 2007 11:46:17 -0500 21, 25 -- Message-ID: {91108EF9255B394CBF8B7E3789814A4101C0B9A0-at-UM-XMAIL08.um.umsystem.edu} 21, 25 -- In-Reply-To: {200705021554.l42FsU8e024507-at-ns.microscopy.com} 21, 25 -- X-MS-Has-Attach: 21, 25 -- X-MS-TNEF-Correlator: 21, 25 -- Thread-Topic: [Microscopy] Re: Help with enhancing contrast of Mitochondrial cristae 21, 25 -- Thread-Index: AceM0i7yopsJkBOARge6j1nHtiiHRQAA61fQ 21, 25 -- References: {200705021554.l42FsU8e024507-at-ns.microscopy.com} 21, 25 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 21, 25 -- To: {microscopy-at-microscopy.com} 21, 25 -- X-OriginalArrivalTime: 02 May 2007 16:46:17.0452 (UTC) FILETIME=[676FCAC0:01C78CD9] 21, 25 -- Content-Transfer-Encoding: 8bit 21, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l42GkHUi002675 ==============================End of - Headers==============================
Hello, This is a posting for one of my friends, so please do not respond to me. Thanks, Vinod
The Rockefeller University, a world-renowned center for research and graduate education in the biomedical sciences, seeks a Manager of Electron Microscopy to join the Bio-Imaging Resource Center: see http://www.rockefeller.edu/bioimaging/. The position will involve the day-to-day management and running of the EM service and the promotion of new technologies offered by the center. The successful applicant will work alongside the Director of the BIRC, who oversees the optical microscopy center directly and provides general oversight of the EM service.
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The Rockefeller University is an Affirmative Action/VEVRAA/Equal Opportunity Employer.
Please feel free to contact Dr. Alison North, the Director of the BIRC, at northa-at-rockefeller.edu for further information or with specific questions.
==============================Original Headers============================== 12, 28 -- From nairvinods-at-gmail.com Wed May 2 13:52:25 2007 12, 28 -- Received: from nz-out-0506.google.com (nz-out-0506.google.com [64.233.162.236]) 12, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l42IqPaL017007 12, 28 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 2 May 2007 13:52:25 -0500 12, 28 -- Received: by nz-out-0506.google.com with SMTP id o37so289873nzf 12, 28 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 02 May 2007 11:52:25 -0700 (PDT) 12, 28 -- DKIM-Signature: a=rsa-sha1; c=relaxed/relaxed; 12, 28 -- d=gmail.com; s=beta; 12, 28 -- h=domainkey-signature:received:received:message-id:date:from:to:subject:cc:mime-version:content-type:content-transfer-encoding:content-disposition; 12, 28 -- b=a1WOSZFq7xRxvcJhCR/BHkZ14TGRhCeuqoHuZ7RjFC1/TCku7X1omWaMRyzT9AcvGTBXrWDDuzIjh74aCUFs24T9T61ppkl3Rqp+2W4ilIXgO9Pn384OV/P1rx1TR0mwb08j7DTtCrmsbePVUG39JSvIzRO6PuRHQuDOgeDUgVM= 12, 28 -- DomainKey-Signature: a=rsa-sha1; c=nofws; 12, 28 -- d=gmail.com; s=beta; 12, 28 -- h=received:message-id:date:from:to:subject:cc:mime-version:content-type:content-transfer-encoding:content-disposition; 12, 28 -- b=SCh47pZOkas5EfnZQCIv/j4Rg8jB2Tc1yFlk3x6quNWQ2TtqI+ayzty1ywPSA5yblzWhCUZdfWCK+XQeneOYG/MXoBEGm3Er+ifhZrdtmSB1ZU2+ywp6tBFnkpSaRoIAWgaDdHiR0BYl3/FQpMjOQUrs0ozslYwUPl3LAqg0iyo= 12, 28 -- Received: by 10.114.136.1 with SMTP id j1mr330498wad.1178131944847; 12, 28 -- Wed, 02 May 2007 11:52:24 -0700 (PDT) 12, 28 -- Received: by 10.114.199.20 with HTTP; Wed, 2 May 2007 11:52:24 -0700 (PDT) 12, 28 -- Message-ID: {ea42a3900705021152x20fb4e29sc200bbbbcc4fc3bb-at-mail.gmail.com} 12, 28 -- Date: Wed, 2 May 2007 12:52:24 -0600 12, 28 -- From: "Vinod Nair" {nairvinods-at-gmail.com} 12, 28 -- To: Microscopy-at-MSA.Microscopy.Com 12, 28 -- Subject: "Rockefeller University seeks Manager for Electron Microscopy unit" 12, 28 -- Cc: northa-at-mail.rockefeller.edu 12, 28 -- MIME-Version: 1.0 12, 28 -- Content-Type: text/plain; charset=UTF-8; format=flowed 12, 28 -- Content-Disposition: inline 12, 28 -- Content-Transfer-Encoding: 8bit 12, 28 -- X-MIME-Autoconverted: from base64 to 8bit by ns.microscopy.com id l42IqPaL017007 ==============================End of - Headers==============================
Randy, I had good luck with a Hotpack, model # H-1125 in another lab. Very reliable and good manufacturer support if needed. Measure your space very carefully! Some other units were 3/4" too tall to fit under the lab top. Larry
TindallR-at-missouri.edu wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Dear Listers, } } Does anyone have a recommendation for a laboratory glassware washer that } will fit under a counter (about like a home dishwasher size), handle } light duty of two or three loads a week, wash labware to a level } acceptable for EM use, and, ahem, not break the bank? Does something } like this exist for $5000 or thereabouts? Am I delusional? (Don't } answer that......) } } We currently have users privileges on a really nice huge heavy duty lab } dishwasher, but the day is coming when this will end. Also, we don't } always have access exactly when we most need it. We want our own! } } Thanks in advance. Vendor replies are most welcome. } } Cheers, } Randy } } Randy Tindall } Senior EM Specialist } Electron Microscopy Core Facility---We Do Small Well! } W125 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-2227 } Email: tindallr-at-missouri.edu } Web: http://www.emc.missouri.edu } On-line calendar: } http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount= } Week&NavType=Both&Type=TimePlan } } } ==============================Original Headers============================== } 7, 23 -- From TindallR-at-missouri.edu Mon Apr 30 17:02:38 2007 } 7, 23 -- Received: from um-nsmtpout1.um.umsystem.edu (um-nsmtpout1.um.umsystem.edu [209.106.228.53]) } 7, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l3UM2caD011664 } 7, 23 -- for {microscopy-at-microscopy.com} ; Mon, 30 Apr 2007 17:02:38 -0500 } 7, 23 -- Received: from UM-XMAIL08.um.umsystem.edu ([209.106.228.34]) by um-nsmtpout1.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); } 7, 23 -- Mon, 30 Apr 2007 17:02:38 -0500 } 7, 23 -- x-mimeole: Produced By Microsoft Exchange V6.5 } 7, 23 -- Content-class: urn:content-classes:message } 7, 23 -- MIME-Version: 1.0 } 7, 23 -- Content-Type: text/plain; } 7, 23 -- charset="us-ascii" } 7, 23 -- Subject: Labware washers } 7, 23 -- Date: Mon, 30 Apr 2007 17:02:37 -0500 } 7, 23 -- Message-ID: {91108EF9255B394CBF8B7E3789814A4101C0B995-at-UM-XMAIL08.um.umsystem.edu} } 7, 23 -- X-MS-Has-Attach: } 7, 23 -- X-MS-TNEF-Correlator: } 7, 23 -- Thread-Topic: Labware washers } 7, 23 -- Thread-Index: AceLc06uPqd+W3UtSgOAFud+EFqhWQ== } 7, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} } 7, 23 -- To: {microscopy-at-microscopy.com} } 7, 23 -- X-OriginalArrivalTime: 30 Apr 2007 22:02:38.0419 (UTC) FILETIME=[44260230:01C78B73] } 7, 23 -- Content-Transfer-Encoding: 8bit } 7, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l3UM2caD011664 } ==============================End of - Headers============================== }
-- Larry Ackerman, Associate Specialist UCSF, Dept. of Anatomy, Rm S1347 513 Parnassus Ave., Box 0452 San Francisco, CA 94143
larry.ackerman-at-ucsf.edu
==============================Original Headers============================== 5, 28 -- From Larry.Ackerman-at-ucsf.edu Wed May 2 14:24:08 2007 5, 28 -- Received: from emfmcb02.ucsfmedicalcenter.org (emfmcb02.ucsfmedicalcenter.org [64.54.46.98]) 5, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l42JO8aq029045 5, 28 -- for {Microscopy-at-microscopy.com} ; Wed, 2 May 2007 14:24:08 -0500 5, 28 -- Received: from 64.54.128.152 by emfmcb01.ucsfmedicalcenter.org with 5, 28 -- ESMTP (Tumbleweed Email Firewall SMTP Relay (Email Firewall v6.1.0)); 5, 28 -- Wed, 02 May 2007 12:34:51 -0700 5, 28 -- X-Server-Uuid: E2E48A14-EE5B-4280-A138-188440602EDD 5, 28 -- Received: from baluk.ucsf.edu ([128.218.123.88]) by exvs06.net.ucsf.edu 5, 28 -- with Microsoft SMTPSVC(6.0.3790.3959); Wed, 2 May 2007 12:24:00 -0700 5, 28 -- Message-ID: {4638E54F.2080606-at-ucsf.edu} 5, 28 -- Date: Wed, 02 May 2007 12:23:59 -0700 5, 28 -- From: "Larry Ackerman" {larry.ackerman-at-ucsf.edu} 5, 28 -- Reply-to: larry.ackerman-at-ucsf.edu 5, 28 -- Organization: UCSF, NeuroAnatomy 5, 28 -- User-Agent: Thunderbird 2.0.0.0 (Macintosh/20070326) 5, 28 -- MIME-Version: 1.0 5, 28 -- To: TindallR-at-missouri.edu, Microscopy-at-microscopy.com 5, 28 -- Subject: Re: [Microscopy] Labware washers 5, 28 -- References: {200704302207.l3UM74eH025435-at-ns.microscopy.com} 5, 28 -- In-Reply-To: {200704302207.l3UM74eH025435-at-ns.microscopy.com} 5, 28 -- X-OriginalArrivalTime: 02 May 2007 19:24:00.0078 (UTC) 5, 28 -- FILETIME=[6F99EEE0:01C78CEF] 5, 28 -- X-WSS-ID: 6A2638510MC1288805-01-01 5, 28 -- Content-Type: text/plain; 5, 28 -- charset=iso-8859-1; 5, 28 -- format=flowed 5, 28 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Sorry if this is a double post. I am looking for 2X and 4X CFI Plan Achromat Objectives for a Nikon Eclipse ME600. 1X Achromat and/or 2X or 4X Apochromats would be great if they were affordable (under $1,000 each) but I won't hold my breath waiting for that. Does anyone have or know of these objectives, new or used, in stock and for sale anywhere? Nikon USA does not have them in stock, neither does our Nikon dealer, and we need to get at least one of these objectives fairly soon - within a week if possible.
The low resolution is so that I can increase the field of view for digital image analysis. If anyone knows of another way to accomplish this, or knows of third-party suppliers that make objectives that fit the Nikon, I'd love to hear about it.
Robert Zonis Product Development Chemist, LMTC Sanford L.P. - A Newell Rubbermaid Company 3 Sharpie Way Shelbyville, TN 37160 Direct: +1 (931) 685-6635 Fax: +1 (931) 685-6613 robert.zonis-at-sanford.com www.papermate.com
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==============================Original Headers============================== 8, 34 -- From Robert.Zonis-at-sanford.com Wed May 2 15:14:46 2007 8, 34 -- Received: from mail47.messagelabs.com (mail47.messagelabs.com [216.82.240.163]) 8, 34 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l42KEk1w009373 8, 34 -- for {Microscopy-at-microscopy.com} ; Wed, 2 May 2007 15:14:46 -0500 8, 34 -- X-VirusChecked: Checked 8, 34 -- X-Env-Sender: Robert.Zonis-at-sanford.com 8, 34 -- X-Msg-Ref: server-13.tower-47.messagelabs.com!1178136823!32295282!1 8, 34 -- X-StarScan-Version: 5.5.10.7.1; banners=sanford.com,-,- 8, 34 -- X-Originating-IP: [12.2.115.11] 8, 34 -- Received: (qmail 25794 invoked from network); 2 May 2007 20:13:43 -0000 8, 34 -- Received: from gnat.newellco.com (HELO nafepncsvpout1.nr.ad.newellco.com) (12.2.115.11) 8, 34 -- by server-13.tower-47.messagelabs.com with SMTP; 2 May 2007 20:13:43 -0000 8, 34 -- Received: from nafepncsefe02.nr.ad.newellco.com (unverified) by nafepncsvpout1.nr.ad.newellco.com 8, 34 -- (Content Technologies SMTPRS 4.3.12) with ESMTP id {T7f59c362de0a0599261014-at-nafepncsvpout1.nr.ad.newellco.com} for {Microscopy-at-microscopy.com} ; 8, 34 -- Wed, 2 May 2007 15:13:43 -0500 8, 34 -- Received: from nashbsasebe01.nr.ad.newellco.com ([10.217.223.198]) by nafepncsefe02.nr.ad.newellco.com with Microsoft SMTPSVC(6.0.3790.1830); 8, 34 -- Wed, 2 May 2007 15:13:43 -0500 8, 34 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 8, 34 -- Content-class: urn:content-classes:message 8, 34 -- MIME-Version: 1.0 8, 34 -- Content-Type: text/plain; 8, 34 -- charset="iso-8859-1" 8, 34 -- Subject: Optical Microscopy - Objectives for Nikon Eclipse ME600L 8, 34 -- Date: Wed, 2 May 2007 15:13:40 -0500 8, 34 -- Message-ID: {8809C39726DC5F43824A3D3BE5D0286A0C6F5D21-at-nashbsasebe01.nr.ad.newellco.com} 8, 34 -- X-MS-Has-Attach: 8, 34 -- X-MS-TNEF-Correlator: 8, 34 -- Thread-Topic: Optical Microscopy - Objectives for Nikon Eclipse ME600L 8, 34 -- Thread-Index: AceM9mBQCovc1cRPRUKyhhd+/48PPw== 8, 34 -- From: "Zonis, Robert" {Robert.Zonis-at-sanford.com} 8, 34 -- To: {Microscopy-at-microscopy.com} 8, 34 -- X-OriginalArrivalTime: 02 May 2007 20:13:43.0347 (UTC) FILETIME=[61C4A830:01C78CF6] 8, 34 -- Content-Transfer-Encoding: 8bit 8, 34 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l42KEk1w009373 ==============================End of - Headers==============================
RE: Gas reaction chemistry in a large-chamber SEM?
I am looking for people who have found creative ways to do gas-reaction chemistry in a "large" chamber SEM, ideally with a heating stage, gas injection and better control of the vacuum/cleanliness (near-UHV). Ideas about chamber within a chamber designs, better cleaning/pumping for the large chamber and/or experience with gas-injection in an environmental SEM would be most welcome! I am familiar with the designs for TEM/STEM, but am looking for ideas based on SEMs. Please feel free to respond either directly or include the distribution. Thank you in advance for your suggestions!
konrad
Disclaimer: i work for the Hitachi electron microscope division so please do NOT disclose any confidential or proprietary information, only open domain discussion!!
==============================Original Headers============================== 6, 26 -- From Konrad.Jarausch-at-hitachi-hta.com Wed May 2 15:46:53 2007 6, 26 -- Received: from mail1.hitachi.net (mail1.hitachi.net [128.241.23.75]) 6, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l42KkrTR021333 6, 26 -- for {Microscopy-at-microscopy.com} ; Wed, 2 May 2007 15:46:53 -0500 6, 26 -- Received: from sjc-hal-smtp02.hal.hitachi.local ([137.168.46.11]) 6, 26 -- by mail1.hitachi.net (8.13.7+Sun/8.13.7) with SMTP id l42KkqAA012322 6, 26 -- for {Microscopy-at-microscopy.com} ; Wed, 2 May 2007 13:46:52 -0700 (PDT) 6, 26 -- Received: from exchange.hitachi-hta.com ([137.168.7.134]) 6, 26 -- by sjc-hal-smtp02.hal.hitachi.local (SMSSMTP 4.1.11.41) with SMTP id M2007050213464715801 6, 26 -- for {Microscopy-at-microscopy.com} ; Wed, 02 May 2007 13:46:47 -0700 6, 26 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 6, 26 -- Content-class: urn:content-classes:message 6, 26 -- MIME-Version: 1.0 6, 26 -- Content-Type: text/plain; 6, 26 -- charset="us-ascii" 6, 26 -- Subject: Gas reaction chemistry in a "large" chamber SEM? 6, 26 -- Date: Wed, 2 May 2007 15:43:25 -0500 6, 26 -- Message-ID: {A7FCF7B158674D4E83DA2203C3B0ABEA0399B273-at-exchange.hitachi-hta.com} 6, 26 -- X-MS-Has-Attach: 6, 26 -- X-MS-TNEF-Correlator: 6, 26 -- Thread-Topic: Gas reaction chemistry in a "large" chamber SEM? 6, 26 -- Thread-Index: AceM+og4yvH+e2R1RCO0xuL+bjjaCQ== 6, 26 -- From: "Jarausch, Konrad" {Konrad.Jarausch-at-hitachi-hta.com} 6, 26 -- To: {Microscopy-at-microscopy.com} 6, 26 -- Content-Transfer-Encoding: 8bit 6, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l42KkrTR021333 ==============================End of - Headers==============================
I've started getting information from the kids about what kind of samples they want to look at using our SEM when it is up and running, and one student gave me a bit of a baffler. I'm generally familiar with various sample prep techniques for different samples of fibers, biologicals, non-organic materials, etc., but this one stumped me.
The student read somewhere that spider webs achieve their elasticity by small bundles of fibers inside the sticky "glue" on the web. They want to see this first hand. Any suggestions on how to prepare a spider web? There is no shortage of them around here.
--Justin A. Kraft Atlantis Academy West Palm Beach, FL
==============================Original Headers============================== 3, 26 -- From kraftpiano-at-gmail.com Wed May 2 18:50:47 2007 3, 26 -- Received: from wr-out-0506.google.com (wr-out-0506.google.com [64.233.184.235]) 3, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l42NolsV003024 3, 26 -- for {microscopy-at-microscopy.com} ; Wed, 2 May 2007 18:50:47 -0500 3, 26 -- Received: by wr-out-0506.google.com with SMTP id i31so361708wra 3, 26 -- for {microscopy-at-microscopy.com} ; Wed, 02 May 2007 16:50:47 -0700 (PDT) 3, 26 -- DKIM-Signature: a=rsa-sha1; c=relaxed/relaxed; 3, 26 -- d=gmail.com; s=beta; 3, 26 -- h=domainkey-signature:received:received:message-id:date:from:to:subject:mime-version:content-type:content-transfer-encoding:content-disposition; 3, 26 -- b=MWb4z9X8hzShbNkjV9OMoe6UChWoRwZU2NRo3GSi4nJPR+t2eH/YA9cpaO4ga6Mkn7r6Hy7x3yW5GyWH6vEjzkRIZdE/tBEAkGcAco1064loRgyD86YSFiE8gqNDzDyJRe/bHYnu9nqDMGwKnOiX3bQadK+mCn+4XNd8rC6WDUE= 3, 26 -- DomainKey-Signature: a=rsa-sha1; c=nofws; 3, 26 -- d=gmail.com; s=beta; 3, 26 -- h=received:message-id:date:from:to:subject:mime-version:content-type:content-transfer-encoding:content-disposition; 3, 26 -- b=ZBtNFVHwLQy6YPOPYvvuNnQpxOuq5DtttgfuqdTE3m2A2QQ2kH9dZPoThqKbBCLroealDQc7zlFmb9kr1GNz3U46+QrOKM+nHnyoZ1O44bz4eT64RiaS+u1I+GFrejJSPX+Ho1UuqiSic03RbVgq0mImCCnj40VIsatcADQHVVk= 3, 26 -- Received: by 10.114.61.1 with SMTP id j1mr432292waa.1178149846592; 3, 26 -- Wed, 02 May 2007 16:50:46 -0700 (PDT) 3, 26 -- Received: by 10.114.79.12 with HTTP; Wed, 2 May 2007 16:50:46 -0700 (PDT) 3, 26 -- Message-ID: {25e2b0d20705021650w611d5d5dqa5860f080e3418d8-at-mail.gmail.com} 3, 26 -- Date: Wed, 2 May 2007 19:50:46 -0400 3, 26 -- From: "Justin Kraft" {kraftpiano-at-gmail.com} 3, 26 -- To: microscopy-at-microscopy.com 3, 26 -- Subject: SEM Sample prep advice. 3, 26 -- MIME-Version: 1.0 3, 26 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 3, 26 -- Content-Transfer-Encoding: 7bit 3, 26 -- Content-Disposition: inline ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.com/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both jgsheridanmicroscopy-at-gmail.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: jgsheridanmicroscopy-at-gmail.com Name: John Sheridan
Organization: TCD
Title-Subject: [Filtered] EDX: ES vision software
Question: Hi all!
Does anyone know who makes "ES vision" , it's a piece of software for EDX analysis. Google has not helped me so far.
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both rra-at-stowers-institute.org as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: rra-at-stowers-institute.org Name: Rhonda Allen
Organization: Stowers Institute
Title-Subject: [Filtered] Mitochondrial Staining
Question: I am sure someone else can elaborate more than I, but Hyatt state that Phosphotungstic Acid after glutaraldehyde fixation in an aqueous acidic medium intensely stains mitochoindrial matix, cisternae of er, and others.
This is from "Principles and Techniques of Electron Microscopy", 4th edition, page 313.
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both tchallman-at-case4n6.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Title-Subject: [Filtered] Hitachi SEM 2460N Service Contract
Question: Once again, we are looking for qualified and experienced people to provide an alternative to Hitachi's SEM Service Contract - annual preventative maintenance and occassional mishaps in the electronics. Do you recommend anyone, particularly in the Pacific Northwest?
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both robert.zonis-at-sanford.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: robert.zonis-at-sanford.com Name: Robert Zonis
Organization: Sanford
Title-Subject: [Filtered] Help with optical objectives
Question: Does anybody know where I can get Nikon CFI Plan Achromat objectives in UW 2x or 4x relatively quickly? I'm doing some image analysis and I need a wider field of view.
Alternatively, does anyone know if there's a third-party supplier of objectves for Nikons?
Robert Zonis Product Development Chemist, LMTC Sanford L.P. ñ A Newell Rubbermaid Company Liquid Manufacturing and Technology Center 3 Sharpie Way Shelbyville, TN 37160 Direct: +1 (931) 685-6635 Fax: +1 (931) 685-6613
Email: jgsheridanmicroscopy-at-gmail.com John Sheridan wrote: ================================================== Title-Subject: [Filtered] EDX: ES vision software
Question: Hi all!
Does anyone know who makes "ES vision" , it's a piece of software for EDX analysis. Google has not helped me so far. ================================================= Are you thinking about the Electron Flight Simulator (EFS) software used in conjuction with EDS, see URL http://www.2spi.com/catalog/software/efs.html
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
==============================Original Headers============================== 7, 20 -- From cgarber-at-2spi.com Wed May 2 19:52:25 2007 7, 20 -- Received: from diskless5.axs2000.net (mail.axs2000.net [209.120.196.43]) 7, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l430qPGr028629 7, 20 -- for {microscopy-at-microscopy.com} ; Wed, 2 May 2007 19:52:25 -0500 7, 20 -- Received: from [192.168.1.103] (c-71-230-36-118.hsd1.pa.comcast.net [71.230.36.118]) 7, 20 -- (authenticated bits=0) 7, 20 -- by diskless5.axs2000.net (8.12.11/8.12.11) with ESMTP id l430qOpp031073 7, 20 -- for {microscopy-at-microscopy.com} ; Wed, 2 May 2007 20:52:24 -0400 7, 20 -- X-IDV-FirstRcvd: c-71-230-36-118.hsd1.pa.comcast.net [71.230.36.118] 7, 20 -- X-IDV-HELO: [192.168.1.103] 7, 20 -- X-IDV-Authenticated-User: cgarber 7, 20 -- Message-ID: {46393241.3080807-at-2spi.com} 7, 20 -- Date: Wed, 02 May 2007 20:52:17 -0400 7, 20 -- From: "Garber, Charles" {cgarber-at-2spi.com} 7, 20 -- User-Agent: Thunderbird 2.0.0.0 (Windows/20070326) 7, 20 -- MIME-Version: 1.0 7, 20 -- To: microscopy-at-microscopy.com 7, 20 -- Subject: ES vision? 7, 20 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 7, 20 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
All the fuss over microprobes was fun but if you want to get your teeth into names used by scientists (?) what about "optical microscopy"?
With my 43 years in the business I have always taught that whilst light microscopes have "light optics" electron microscopes have "electron optics", the principles are the same. So when I see a piece about optical microscopes I always look to see if it should be of interest to me?
What is the general thought?
Steve Chapman Senior Consultant Protrain For electron microscopy consultancy and training world wide Tel +44 1280 816512 Fax +44 1280 814007 Mobile +44 7711 606967 Web www.emcourses.com
==============================Original Headers============================== 6, 26 -- From protrain-at-emcourses.com Thu May 3 06:06:37 2007 6, 26 -- Received: from smtp1.pri.skybb.uk.easynet.net (smtp1.pri.skybb.uk.easynet.net [87.86.189.69]) 6, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l43B6bku028581 6, 26 -- for {microscopy-at-microscopy.com} ; Thu, 3 May 2007 06:06:37 -0500 6, 26 -- Received: from [90.200.252.11] (helo=advent) 6, 26 -- by smtp1.pri.skybb.uk.easynet.net with smtp (Exim 4.62) 6, 26 -- (envelope-from {protrain-at-emcourses.com} ) 6, 26 -- id 1HjZ8u-0002y3-9V 6, 26 -- for microscopy-at-microscopy.com; Thu, 03 May 2007 12:06:36 +0100 6, 26 -- Message-ID: {001901c78d73$bc033230$0200a8c0-at-advent} 6, 26 -- Reply-To: "Steve Chapman" {protrain-at-emcourses.com} 6, 26 -- From: "Steve Chapman" {protrain-at-emcourses.com} 6, 26 -- To: "American Soc" {microscopy-at-microscopy.com} 6, 26 -- Subject: [Microscopy] Optical Microscopy 6, 26 -- Date: Thu, 3 May 2007 12:10:57 +0100 6, 26 -- Organization: Protrain 6, 26 -- MIME-Version: 1.0 6, 26 -- Content-Type: text/plain; 6, 26 -- format=flowed; 6, 26 -- charset="iso-8859-1"; 6, 26 -- reply-type=original 6, 26 -- Content-Transfer-Encoding: 7bit 6, 26 -- X-Priority: 3 6, 26 -- X-MSMail-Priority: Normal 6, 26 -- X-Mailer: Microsoft Outlook Express 6.00.2900.3028 6, 26 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 ==============================End of - Headers==============================
Having worn glasses since eight grade, I have come to associate optical with glass the photons.
Phototonic microscopy? sounds like a title of a paper in search of a little primping Non-modified bright field? - sounds like a title of a paper in search of a little primping from a major university Light microscopy? - sort of suggest their might be a heavy microscopy Refractory microscopy - sound like microscopy in hell Specular microscopy - the study of the stock market in great detail..
Tongue firmly implanted in cheek: Let's call it Bruce. I got the idea from a microscopy society bog ( http://www.msneo.org/2006/06/blog-mail.html) who knows..........
Stay safe..............Frank
protrain-at-emcourse s.com To: frank.karl-at-degussa.com cc: 05/03/2007 07:09 Subject: [Microscopy] Optical Microscopy AM Please respond to protrain
The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
Hi
All the fuss over microprobes was fun but if you want to get your teeth into names used by scientists (?) what about "optical microscopy"?
With my 43 years in the business I have always taught that whilst light microscopes have "light optics" electron microscopes have "electron optics", the principles are the same. So when I see a piece about optical microscopes I always look to see if it should be of interest to me?
What is the general thought?
Steve Chapman Senior Consultant Protrain For electron microscopy consultancy and training world wide Tel +44 1280 816512 Fax +44 1280 814007 Mobile +44 7711 606967 Web www.emcourses.com
==============================Original Headers============================== 6, 26 -- From protrain-at-emcourses.com Thu May 3 06:06:37 2007 6, 26 -- Received: from smtp1.pri.skybb.uk.easynet.net (smtp1.pri.skybb.uk.easynet.net [87.86.189.69]) 6, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l43B6bku028581 6, 26 -- for {microscopy-at-microscopy.com} ; Thu, 3 May 2007 06:06:37 -0500 6, 26 -- Received: from [90.200.252.11] (helo=advent) 6, 26 -- by smtp1.pri.skybb.uk.easynet.net with smtp (Exim 4.62) 6, 26 -- (envelope-from {protrain-at-emcourses.com} ) 6, 26 -- id 1HjZ8u-0002y3-9V 6, 26 -- for microscopy-at-microscopy.com; Thu, 03 May 2007 12:06:36 +0100 6, 26 -- Message-ID: {001901c78d73$bc033230$0200a8c0-at-advent} 6, 26 -- Reply-To: "Steve Chapman" {protrain-at-emcourses.com} 6, 26 -- From: "Steve Chapman" {protrain-at-emcourses.com} 6, 26 -- To: "American Soc" {microscopy-at-microscopy.com} 6, 26 -- Subject: [Microscopy] Optical Microscopy 6, 26 -- Date: Thu, 3 May 2007 12:10:57 +0100 6, 26 -- Organization: Protrain 6, 26 -- MIME-Version: 1.0 6, 26 -- Content-Type: text/plain; 6, 26 -- format=flowed; 6, 26 -- charset="iso-8859-1"; 6, 26 -- reply-type=original 6, 26 -- Content-Transfer-Encoding: 7bit 6, 26 -- X-Priority: 3 6, 26 -- X-MSMail-Priority: Normal 6, 26 -- X-Mailer: Microsoft Outlook Express 6.00.2900.3028 6, 26 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 ==============================End of - Headers==============================
==============================Original Headers============================== 27, 18 -- From frank.karl-at-degussa.com Thu May 3 07:00:41 2007 27, 18 -- Received: from framailout1.rz.itson.com (mailout2.degussa.com [149.216.91.173]) 27, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l43C0f20019657 27, 18 -- for {microscopy-at-msa.microscopy.com} ; Thu, 3 May 2007 07:00:41 -0500 27, 18 -- Received: from mobuscomm01.mail.degussa.com (uss1026.applications.degussanet.com [10.88.88.98]) 27, 18 -- by framailout1.rz.itson.com (8.13.4/8.13.4/Debian-3sarge3) with ESMTP id l43C0ZLo027649; 27, 18 -- Thu, 3 May 2007 14:00:36 +0200 27, 18 -- In-Reply-To: {200705031109.l43B9Mcd030690-at-ns.microscopy.com} 27, 18 -- Subject: Re: [Microscopy] Optical Microscopy 27, 18 -- To: protrain-at-emcourses.com, microscopy-at-msa.microscopy.com 27, 18 -- X-Mailer: Lotus Notes Release 6.5.5 November 30, 2005 27, 18 -- Message-ID: {OFAD8E7B5A.9EF57A57-ON862572D0.0040CD9B-852572D0.0041F460-at-degussa.com} 27, 18 -- From: frank.karl-at-degussa.com 27, 18 -- Date: Thu, 3 May 2007 08:00:31 -0400 27, 18 -- X-MIMETrack: Serialize by Router on MOBUSComm01/DHexternal/US(Release 6.5.1|January 21, 2004) at 27, 18 -- 05/03/2007 07:00:36 AM 27, 18 -- MIME-Version: 1.0 27, 18 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
Light Microscopy? I once got an assignment where the student thought this was a special tool to study the activity of chlorophyll!
Dave
-----Original Message----- X-from: frank.karl-at-degussa.com [mailto:frank.karl-at-degussa.com] Sent: 03 May 2007 13:05 To: David Patton
Guilty!
Having worn glasses since eight grade, I have come to associate optical with glass the photons.
Phototonic microscopy? sounds like a title of a paper in search of a little primping Non-modified bright field? - sounds like a title of a paper in search of a little primping from a major university Light microscopy? - sort of suggest their might be a heavy microscopy Refractory microscopy - sound like microscopy in hell Specular microscopy - the study of the stock market in great detail..
Tongue firmly implanted in cheek: Let's call it Bruce. I got the idea from a microscopy society bog ( http://www.msneo.org/2006/06/blog-mail.html) who knows..........
Stay safe..............Frank
protrain-at-emcourse
s.com To: frank.karl-at-degussa.com
cc:
05/03/2007 07:09 Subject: [Microscopy] Optical Microscopy AM
The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
Hi
All the fuss over microprobes was fun but if you want to get your teeth into names used by scientists (?) what about "optical microscopy"?
With my 43 years in the business I have always taught that whilst light microscopes have "light optics" electron microscopes have "electron optics", the principles are the same. So when I see a piece about optical microscopes I always look to see if it should be of interest to me?
What is the general thought?
Steve Chapman Senior Consultant Protrain For electron microscopy consultancy and training world wide Tel +44 1280 816512 Fax +44 1280 814007 Mobile +44 7711 606967 Web www.emcourses.com
==============================Original Headers============================== 6, 26 -- From protrain-at-emcourses.com Thu May 3 06:06:37 2007 6, 26 -- Received: from smtp1.pri.skybb.uk.easynet.net (smtp1.pri.skybb.uk.easynet.net [87.86.189.69]) 6, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l43B6bku028581 6, 26 -- for {microscopy-at-microscopy.com} ; Thu, 3 May 2007 06:06:37 -0500 6, 26 -- Received: from [90.200.252.11] (helo=advent) 6, 26 -- by smtp1.pri.skybb.uk.easynet.net with smtp (Exim 4.62) 6, 26 -- (envelope-from {protrain-at-emcourses.com} ) 6, 26 -- id 1HjZ8u-0002y3-9V 6, 26 -- for microscopy-at-microscopy.com; Thu, 03 May 2007 12:06:36 +0100 6, 26 -- Message-ID: {001901c78d73$bc033230$0200a8c0-at-advent} 6, 26 -- Reply-To: "Steve Chapman" {protrain-at-emcourses.com} 6, 26 -- From: "Steve Chapman" {protrain-at-emcourses.com} 6, 26 -- To: "American Soc" {microscopy-at-microscopy.com} 6, 26 -- Subject: [Microscopy] Optical Microscopy 6, 26 -- Date: Thu, 3 May 2007 12:10:57 +0100 6, 26 -- Organization: Protrain 6, 26 -- MIME-Version: 1.0 6, 26 -- Content-Type: text/plain; 6, 26 -- format=flowed; 6, 26 -- charset="iso-8859-1"; 6, 26 -- reply-type=original 6, 26 -- Content-Transfer-Encoding: 7bit 6, 26 -- X-Priority: 3 6, 26 -- X-MSMail-Priority: Normal 6, 26 -- X-Mailer: Microsoft Outlook Express 6.00.2900.3028 6, 26 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 ==============================End of - Headers==============================
==============================Original Headers============================== 27, 18 -- From frank.karl-at-degussa.com Thu May 3 07:00:41 2007 27, 18 -- Received: from framailout1.rz.itson.com (mailout2.degussa.com [149.216.91.173]) 27, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l43C0f20019657 27, 18 -- for {microscopy-at-msa.microscopy.com} ; Thu, 3 May 2007 07:00:41 -0500 27, 18 -- Received: from mobuscomm01.mail.degussa.com (uss1026.applications.degussanet.com [10.88.88.98]) 27, 18 -- by framailout1.rz.itson.com (8.13.4/8.13.4/Debian-3sarge3) with ESMTP id l43C0ZLo027649; 27, 18 -- Thu, 3 May 2007 14:00:36 +0200 27, 18 -- In-Reply-To: {200705031109.l43B9Mcd030690-at-ns.microscopy.com} 27, 18 -- Subject: Re: [Microscopy] Optical Microscopy 27, 18 -- To: protrain-at-emcourses.com, microscopy-at-msa.microscopy.com 27, 18 -- X-Mailer: Lotus Notes Release 6.5.5 November 30, 2005 27, 18 -- Message-ID: {OFAD8E7B5A.9EF57A57-ON862572D0.0040CD9B-852572D0.0041F460-at-degussa.com} 27, 18 -- From: frank.karl-at-degussa.com 27, 18 -- Date: Thu, 3 May 2007 08:00:31 -0400 27, 18 -- X-MIMETrack: Serialize by Router on MOBUSComm01/DHexternal/US(Release 6.5.1|January 21, 2004) at 27, 18 -- 05/03/2007 07:00:36 AM 27, 18 -- MIME-Version: 1.0 27, 18 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
This incoming email to UWE has been independently scanned for viruses by McAfee anti-virus software and none were detected
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==============================Original Headers============================== 47, 34 -- From David.Patton-at-uwe.ac.uk Thu May 3 07:33:53 2007 47, 34 -- Received: from mailapp03.uwe.ac.uk (mailapp03.uwe.ac.uk [164.11.132.65]) 47, 34 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l43CXqts031605 47, 34 -- for {microscopy-at-microscopy.com} ; Thu, 3 May 2007 07:33:52 -0500 47, 34 -- Received: from (mta01.uwe.ac.uk [164.11.132.60]) by mailapp03.uwe.ac.uk with smtp 47, 34 -- id 0fa7_4d0aeb50_f972_11db_9046_00142221cca9; 47, 34 -- Thu, 03 May 2007 13:32:19 +0100 47, 34 -- Received: from egen-uwe01.campus.ads.uwe.ac.uk 47, 34 -- (egen-uwe01.uwe.ac.uk [164.11.249.121]) 47, 34 -- by mta01.uwe.ac.uk (iPlanet Messaging Server 5.2 HotFix 2.07 (built Jun 24 47, 34 -- 2005)) with ESMTP id {0JHG005I4TJZBU-at-mta01.uwe.ac.uk} for 47, 34 -- microscopy-at-microscopy.com; Thu, 03 May 2007 13:33:36 +0100 (BST) 47, 34 -- Date: Thu, 03 May 2007 13:31:31 +0100 47, 34 -- From: David Patton {David.Patton-at-uwe.ac.uk} 47, 34 -- Subject: RE: [Microscopy] Re: Optical Microscopy 47, 34 -- In-reply-to: {200705031204.l43C4Vio026439-at-ns.microscopy.com} 47, 34 -- To: frank.karl-at-degussa.com 47, 34 -- Cc: microscopy-at-microscopy.com 47, 34 -- Message-id: {F247F674896BE243AD8263C5280E2BDB02F5B8B7-at-egen-uwe01} 47, 34 -- MIME-version: 1.0 47, 34 -- X-MIMEOLE: Produced By Microsoft Exchange V6.5 47, 34 -- Content-type: text/plain; charset=us-ascii 47, 34 -- Content-class: urn:content-classes:message 47, 34 -- Thread-topic: [Microscopy] Re: Optical Microscopy 47, 34 -- Thread-index: AceNezxZNBI1VVz0SSOoANIrMaL7TQAA4QYA 47, 34 -- X-MS-Has-Attach: 47, 34 -- X-MS-TNEF-Correlator: 47, 34 -- X-NAIMIME-Disclaimer: 1 47, 34 -- X-NAIMIME-Modified: 1 47, 34 -- X-NAI-Spam-Score: -2.5 47, 34 -- X-NAI-Spam-Rules: 1 Rules triggered 47, 34 -- BAYES_00=-2.5 47, 34 -- Content-Transfer-Encoding: 8bit 47, 34 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l43CXqts031605 ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both gas19-at-daimlerchrysler.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Title-Subject: [Filtered] Re: [Microscopy] viaWWW: Help with optical objectives
Question: We have bought two low magnfiication objectives for one of our Nikon microscopes. A 1.5x CF Plan BF EPI Achromat na 0.045 wd 3.6mm for roughly $3k, and a 2.5x CF BF Plan EPI Achromat na 0.075 wd 8.8 mm for roughly $1k. Both have Nikon Japan on the lens. You may try some of the Nikon distributors. Our local distributor is Mager Scientific in Dexter, MI. www.magersci.com. (734) 426-3885.
Gerald Shulke DaimlerChrysler Material Characterization Labs
Steve, Is it ever? I mean of interest to you? More specifically, about electrons rather than photons? My feeling is that despite folks talking about 'electron optics' that the term optical microscopy always means light. Probably the irrationality of names but one could suggest that optics means something different than optical. Optics originally referred to photons but once it was discovered that electrons (and refrigerators) also have wave properties, the term optics was expanded. Wheras optical seems to have retained its photon orientation.
My second order guess.
Tobias
} } Hi } } All the fuss over microprobes was fun but if you want to get your teeth into } names used by scientists (?) what about "optical microscopy"? } } With my 43 years in the business I have always taught that whilst light } microscopes have "light optics" electron microscopes have "electron optics", } the principles are the same. So when I see a piece about optical } microscopes I always look to see if it should be of interest to me? } } What is the general thought? } } Steve Chapman } Senior Consultant Protrain } For electron microscopy consultancy and training world wide } Tel +44 1280 816512 Fax +44 1280 814007 } Mobile +44 7711 606967 Web www.emcourses.com }
Glossary of Microscopical Terms and Definitions, 2nd Ed., NY Microscopical Society, 1989.
Optical microscope. Very ambiguous term since all microscopes involve optics; better to specify light, acoustic, x-ray or electron microscope, etc.
Also, for Frank, from the foreword:
"Words, like eyeglasses, blur everything that they do not make clear." - Joseph Joubert (1754-1824)
Tony
Ps Microprobe is not in the glossary
Pps Glasses since 4th grade.
...................................................................... Andrew Anthony "Tony" Havics, CHMM, CIH, PE pH2, LLC 5250 E US 36, Suite 830 Avon, IN 46123 (317) 718-7020 off (317) 718-7038 fax (317) 409-3238 cell
90% of Risk Management is knowing where to place the decimal point...any consultant can give you the other 10%(SM)
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-----Original Message----- X-from: protrain-at-emcourses.com [mailto:protrain-at-emcourses.com] Sent: Thursday, May 03, 2007 7:13 AM To: ph2-at-sprynet.com
Hi
All the fuss over microprobes was fun but if you want to get your teeth into
names used by scientists (?) what about "optical microscopy"?
With my 43 years in the business I have always taught that whilst light microscopes have "light optics" electron microscopes have "electron optics",
the principles are the same. So when I see a piece about optical microscopes I always look to see if it should be of interest to me?
What is the general thought?
Steve Chapman Senior Consultant Protrain For electron microscopy consultancy and training world wide Tel +44 1280 816512 Fax +44 1280 814007 Mobile +44 7711 606967 Web www.emcourses.com
==============================Original Headers============================== 6, 26 -- From protrain-at-emcourses.com Thu May 3 06:06:37 2007 6, 26 -- Received: from smtp1.pri.skybb.uk.easynet.net (smtp1.pri.skybb.uk.easynet.net [87.86.189.69]) 6, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l43B6bku028581 6, 26 -- for {microscopy-at-microscopy.com} ; Thu, 3 May 2007 06:06:37 -0500 6, 26 -- Received: from [90.200.252.11] (helo=advent) 6, 26 -- by smtp1.pri.skybb.uk.easynet.net with smtp (Exim 4.62) 6, 26 -- (envelope-from {protrain-at-emcourses.com} ) 6, 26 -- id 1HjZ8u-0002y3-9V 6, 26 -- for microscopy-at-microscopy.com; Thu, 03 May 2007 12:06:36 +0100 6, 26 -- Message-ID: {001901c78d73$bc033230$0200a8c0-at-advent} 6, 26 -- Reply-To: "Steve Chapman" {protrain-at-emcourses.com} 6, 26 -- From: "Steve Chapman" {protrain-at-emcourses.com} 6, 26 -- To: "American Soc" {microscopy-at-microscopy.com} 6, 26 -- Subject: [Microscopy] Optical Microscopy 6, 26 -- Date: Thu, 3 May 2007 12:10:57 +0100 6, 26 -- Organization: Protrain 6, 26 -- MIME-Version: 1.0 6, 26 -- Content-Type: text/plain; 6, 26 -- format=flowed; 6, 26 -- charset="iso-8859-1"; 6, 26 -- reply-type=original 6, 26 -- Content-Transfer-Encoding: 7bit 6, 26 -- X-Priority: 3 6, 26 -- X-MSMail-Priority: Normal 6, 26 -- X-Mailer: Microsoft Outlook Express 6.00.2900.3028 6, 26 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 ==============================End of - Headers==============================
==============================Original Headers============================== 28, 27 -- From ph2-at-sprynet.com Thu May 3 10:36:28 2007 28, 27 -- Received: from elasmtp-junco.atl.sa.earthlink.net (elasmtp-junco.atl.sa.earthlink.net [209.86.89.63]) 28, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l43FaSKm010739 28, 27 -- for {microscopy-at-microscopy.com} ; Thu, 3 May 2007 10:36:28 -0500 28, 27 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 28, 27 -- s=dk20050327; d=sprynet.com; 28, 27 -- b=FKDvFEjWUZm2MzJKASacqWpXOIKoH3AbiNCPDPHxe5XUxXGojOoRsRTqp+Inpn6+; 28, 27 -- h=Received:From:To:Cc:Subject:Date:MIME-Version:Content-Type:Content-Transfer-Encoding:X-Mailer:X-MimeOLE:In-reply-to:Thread-Index:Message-ID:X-ELNK-Trace:X-Originating-IP; 28, 27 -- Received: from [75.61.18.94] (helo=user915fa8f284) 28, 27 -- by elasmtp-junco.atl.sa.earthlink.net with asmtp (Exim 4.34) 28, 27 -- id 1HjdM3-0002Sb-5G; Thu, 03 May 2007 11:36:27 -0400 28, 27 -- From: "Tony Havics" {ph2-at-sprynet.com} 28, 27 -- To: {protrain-at-emcourses.com} 28, 27 -- Cc: "Micrscopy Listserve" {microscopy-at-microscopy.com} 28, 27 -- Subject: RE: [Microscopy] Optical Microscopy 28, 27 -- Date: Thu, 3 May 2007 11:36:22 -0400 28, 27 -- MIME-Version: 1.0 28, 27 -- Content-Type: text/plain; 28, 27 -- charset="us-ascii" 28, 27 -- Content-Transfer-Encoding: 7bit 28, 27 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 28, 27 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 28, 27 -- In-reply-to: {200705031113.l43BDRkt004833-at-ns.microscopy.com} 28, 27 -- Thread-Index: AceNdBKIedJssng0TcqQV9jVxjZ8pwAI/iEg 28, 27 -- Message-ID: {E1HjdM3-0002Sb-5G-at-elasmtp-junco.atl.sa.earthlink.net} 28, 27 -- X-ELNK-Trace: 6888e50b2be9b4fee5331016acda17f9774f62a7fc2927845e52b0dd316ca299350badd9bab72f9c350badd9bab72f9c350badd9bab72f9c350badd9bab72f9c 28, 27 -- X-Originating-IP: 75.61.18.94 ==============================End of - Headers==============================
You may be referring to ES Vision developed by Emispec several years ago at the State University of Arizona, and now part of the FEI organisation.
Try: ESVisionSupport-at-FEICO.com
This is from a communication I received from them last year.
Good Luck,
Donald Robertson
///////////////////////////////////////////////// From: ESVisionSupport-at-FEICO.com Subject: RE: Support service for ES Vision Date: July 28, 2006 1:06:43 PM CDT To: donald-at-uwm.edu
Hi Donald,
The support email address you used for this message is the best way to obtain support for your ES Vision system. Please forward your questions or problems to this address and we will respond as soon as possible.
Best Regards,
ES Vision Support
/////////////////////////////////////
On May 2, 2007, at 7:01 PM, jgsheridanmicroscopy-at-gmail.com wrote:
} ---------------------------------------------------------------------- } ------ } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } This Question/Comment was submitted to the Microscopy Listserver } using the WWW based Form at http://www.microscopy.com/MLFormMail.html } ---------------------------------------------------------------------- } ----- } Remember this posting is most likely not from a Subscriber, so when } replying } please copy both jgsheridanmicroscopy-at-gmail.com as well as the } MIcroscopy Listserver } ---------------------------------------------------------------------- } ----- } } Email: jgsheridanmicroscopy-at-gmail.com } Name: John Sheridan } } Organization: TCD } } Title-Subject: [Filtered] EDX: ES vision software } } Question: Hi all! } } Does anyone know who makes "ES vision" , it's a piece of software } for EDX analysis. Google has not helped me so far. } } Many thanks, } } John
Donald Robertson Sr. Instrumentation Specialist HRTEM Lab Department of Physics College of Letters and Science University of Wisconsin - Milwaukee tel: (414) 229 2753
==============================Original Headers============================== 18, 18 -- From donald-at-uwm.edu Thu May 3 10:45:22 2007 18, 18 -- Received: from mail02.imt.uwm.edu (mail02.imt.uwm.edu [129.89.7.44]) 18, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l43FjLjE022376 18, 18 -- for {microscopy-at-microscopy.com} ; Thu, 3 May 2007 10:45:21 -0500 18, 18 -- Received: from [129.89.57.171] (donald.phys.uwm.edu [129.89.57.171]) 18, 18 -- by mail02.imt.uwm.edu (8.13.1/8.13.1) with ESMTP id l43FjJic012532; 18, 18 -- Thu, 3 May 2007 10:45:19 -0500 18, 18 -- In-Reply-To: {200705030001.l4301Xtf024561-at-ns.microscopy.com} 18, 18 -- References: {200705030001.l4301Xtf024561-at-ns.microscopy.com} 18, 18 -- Mime-Version: 1.0 (Apple Message framework v752.2) 18, 18 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 18, 18 -- Message-Id: {7FA5B24E-2F4A-4ED9-99AC-8EB5EAC81353-at-uwm.edu} 18, 18 -- Content-Transfer-Encoding: 7bit 18, 18 -- From: Donald Robertson {donald-at-uwm.edu} 18, 18 -- Subject: Re: [Microscopy] viaWWW: ES vision software 18, 18 -- Date: Thu, 3 May 2007 10:45:13 -0500 18, 18 -- To: jgsheridanmicroscopy-at-gmail.com 18, 18 -- X-Mailer: Apple Mail (2.752.2) ==============================End of - Headers==============================
ASTM standard E175(2005) Standard Terminology of Microscopy
} re: Optical microscope. } Very ambiguous term since all microscopes involve } optics; better to specify light, acoustic, x-ray or } electron microscope, etc. } } }
regards,
JQuinn
==============================Original Headers============================== 7, 12 -- From jquinn-at-www.matscieng.sunysb.edu Thu May 3 11:21:13 2007 7, 12 -- Received: from www.matscieng.sunysb.edu (www.matscieng.sunysb.edu [129.49.36.33]) 7, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l43GLCWA002036 7, 12 -- for {microscopy-at-microscopy.com} ; Thu, 3 May 2007 11:21:13 -0500 7, 12 -- Received: (from jquinn-at-localhost) 7, 12 -- by www.matscieng.sunysb.edu (8.11.6/8.11.6) id l43GLGN18952 7, 12 -- for microscopy-at-microscopy.com; Thu, 3 May 2007 12:21:16 -0400 7, 12 -- Date: Thu, 3 May 2007 12:21:16 -0400 7, 12 -- From: Jim Quinn {jquinn-at-www.matscieng.sunysb.edu} 7, 12 -- Message-Id: {200705031621.l43GLGN18952-at-www.matscieng.sunysb.edu} 7, 12 -- To: microscopy-at-microscopy.com 7, 12 -- Subject: re: optical microscopy ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both tomw-at-uidaho.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: tomw-at-uidaho.edu Name: Tom Williams
Organization: Univ. of Idaho
Title-Subject: [Filtered] Simulated EDS Spectrum
Question: One of our Geology Faculty asked me if I had software that could simulate EDS spectrums for minerals (such as a feldspars or quartz) he could use to generate a number of patterns for educational purposes (without having to actually run each sample on the SEM/EDS-itís a time-constraint issue). I have never seen software that can do this, and told him it is far from trivial. As far as I can determine, my Noran System Six software canít do this. Is anyone aware of a program that can do this??
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both jvtaylo-at-emory.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: jvtaylo-at-emory.edu Name: Jeannette Taylor
Organization: IM&MF / Emory University
Title-Subject: [Filtered] electron diffraction
Question: We are beginning to explore the use of electron diffraction in the study of crystalized biological molucules. We don't have the background here. Some of these molecules are very fragile and disappear before a pattern can be captured on film. An electron diffractin pattern is seen, briefly, then it disappears. We have been following what the micropscope manual instructs on generating electron diffractions and have gotten some advice from a metallurgist. But we need some more help. Using purchased standards, we have gotten some very nice diffraction patterns.
You can use a free Monte Carlo program to simulate the EDS spectrum like WinX-Ray: http://montecarlomodeling.mcgill.ca/download/download.html
If you have a mac with a PowerPC CPU you can use DTSA software: http://www.cstl.nist.gov/div837/Division/outputs/DTSA/DTSA.htm
I know a couple of other software, but you need to do some coding to used them.
A nice list of Monte Carlo softwares can be found on the NISTMonte web page by N. Ritchie: http://www.cstl.nist.gov/div837/837.02/epq/index.html
Usual disclaimer: I am the author of WinX-Ray and I have use and test most of the free softwares mentioned in this post.
Good luck, Hendrix
On 5/3/07, tomw-at-uidaho.edu {tomw-at-uidaho.edu} wrote: } Email: tomw-at-uidaho.edu } Name: Tom Williams } } Organization: Univ. of Idaho } } Title-Subject: [Filtered] Simulated EDS Spectrum } } Question: One of our Geology Faculty asked me if I had software that could simulate EDS spectrums for minerals (such as a feldspars or quartz) he could use to generate a number of patterns for educational purposes (without having to actually run each sample on the SEM/EDS-itís a time-constraint issue). I have never seen software that can do this, and told him it is far from trivial. As far as I can determine, my Noran System Six software canít do this. Is anyone aware of a program that can do this?? } } Thanks } Tom } } } --------------------------------------------------------------------------- } }
==============================Original Headers============================== 9, 30 -- From drix00-at-gmail.com Thu May 3 19:30:40 2007 9, 30 -- Received: from wx-out-0506.google.com (wx-out-0506.google.com [66.249.82.233]) 9, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l440Ue35023693 9, 30 -- for {Microscopy-at-microscopy.com} ; Thu, 3 May 2007 19:30:40 -0500 9, 30 -- Received: by wx-out-0506.google.com with SMTP id t8so627031wxc 9, 30 -- for {Microscopy-at-microscopy.com} ; Thu, 03 May 2007 17:30:40 -0700 (PDT) 9, 30 -- DKIM-Signature: a=rsa-sha1; c=relaxed/relaxed; 9, 30 -- d=gmail.com; s=beta; 9, 30 -- h=domainkey-signature:received:received:message-id:date:from:to:subject:cc:in-reply-to:mime-version:content-type:content-transfer-encoding:content-disposition:references; 9, 30 -- b=Gy0IZmihziI/3iSC3SMdEQAZJ6ZGkgONjtQER+VeyDPMQIxcWvjGlDZVINPafUxVbuhKT0BKJ+CG2giHl1tx/jxXGJilE1YfLL8yYttCa+FbZN+tJghN8bKYHGqNNrNkTxOfPoO+n2fj3eIQu6bqlETtvKbvNHFyTNQvVm+UUFs= 9, 30 -- DomainKey-Signature: a=rsa-sha1; c=nofws; 9, 30 -- d=gmail.com; s=beta; 9, 30 -- h=received:message-id:date:from:to:subject:cc:in-reply-to:mime-version:content-type:content-transfer-encoding:content-disposition:references; 9, 30 -- b=SZPHyhDVnYrrgJpwkhXDSg7htJ93kFxRFNuG6aMJg7+pFOH0FjCNQzQpme4qeHNalA5mWaxFN4FdVwI9q0K+DIr4GvRcgtkxIK9HcEEhqpIvNfgW6Ogvqhu60Ozar0ffeGGu5ENAA92tX6dHvLLebYgfD5WLxNByqvIWT983VnA= 9, 30 -- Received: by 10.90.66.9 with SMTP id o9mr2719695aga.1178238639929; 9, 30 -- Thu, 03 May 2007 17:30:39 -0700 (PDT) 9, 30 -- Received: by 10.90.115.10 with HTTP; Thu, 3 May 2007 17:30:39 -0700 (PDT) 9, 30 -- Message-ID: {a779eeae0705031730j32a2342eu70ec4c475e44a247-at-mail.gmail.com} 9, 30 -- Date: Thu, 3 May 2007 20:30:39 -0400 9, 30 -- From: drix {drix00-at-gmail.com} 9, 30 -- To: tomw-at-uidaho.edu 9, 30 -- Subject: Re: [Microscopy] viaWWW: Simulated EDS Spectrum 9, 30 -- Cc: Microscopy-at-microscopy.com 9, 30 -- In-Reply-To: {200705032348.l43NmZCv017979-at-ns.microscopy.com} 9, 30 -- MIME-Version: 1.0 9, 30 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 9, 30 -- Content-Disposition: inline 9, 30 -- References: {200705032348.l43NmZCv017979-at-ns.microscopy.com} 9, 30 -- Content-Transfer-Encoding: 8bit 9, 30 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l440Ue35023693 ==============================End of - Headers==============================
First, you need a Mac. It might have to be an older Mac.
Then visit the following website, http://www.cstl.nist.gov/div837/Division/outputs/DTSA/DTSA.htm
There you will see a complete description of DTSA which stands for Desktop Spectrum Analyzer.
It will do exactly what you want to do.
You might want to look at my article in Microscopy Today, March 2006, on page 34 to show examples of spectra generated by DTSA.
-Scott
Scott D. Walck, Ph.D. Technical Director South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
US Toll Free: 1-800-728-2233 Tel: (949) 492-2600 Fax: (949) 492-1499
www.southbaytech.com walck-at-southbaytech.com
-----Original Message----- X-from: tomw-at-uidaho.edu [mailto:tomw-at-uidaho.edu] Sent: Thursday, May 03, 2007 4:45 PM To: Walck-at-SouthBayTech.com
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying
please copy both tomw-at-uidaho.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: tomw-at-uidaho.edu Name: Tom Williams
Organization: Univ. of Idaho
Title-Subject: [Filtered] Simulated EDS Spectrum
Question: One of our Geology Faculty asked me if I had software that could simulate EDS spectrums for minerals (such as a feldspars or quartz) he could use to generate a number of patterns for educational purposes (without having to actually run each sample on the SEM/EDS-itís a time-constraint issue). I have never seen software that can do this, and told him it is far from trivial. As far as I can determine, my Noran System Six software canít do this. Is anyone aware of a program that can do this??
On May 3, 2007, at 4:41 PM, jvtaylo-at-emory.edu wrote:
} We are beginning to explore the use of electron diffraction in the } study of crystalized biological molucules. We don't have the } background here. Some of these molecules are very fragile and } disappear before a pattern can be captured on film. An electron } diffractin pattern is seen, briefly, then it disappears. We have been } following what the micropscope manual instructs on generating electron } diffractions and have gotten some advice from a metallurgist. But we } need some more help. Using purchased standards, we have gotten some } very nice diffraction patterns.
Dear Jeannette, You will need to work in low dose conditions. If you are using film, I highly recommend the most sensitive film available, and such films as LoDose or MRF32 X-ray films are about 20 times more sensitive than SO163. The only problem with these is that they must be handled in total darkness, so it will be difficult to cut them to size. Loading and unloading the camera, loading the racks, and developing the film are also somewhat difficult, but the skill to do that will come after a few times doing those tasks. Another concern is static electricity, which will make very interesting patterns on the developed film, but these will destroy the ED data. The simplest way to get good data is to insert the selected area aperture that is the right size for the crystals you are looking at, put the scope in diffraction mode, use a very small condenser aperture, a high spot size, and underfocus the beam until you have parallel illumination, then scan the grid in diffraction mode, either looking for a good pattern or defocussing the beam and looking for the distorted image of the crystal in the zero-order spot. Blank the beam, set the lenses to the proper values to obtain a spot pattern (if you were searching in defocussed diffraction) start the exposure, and turn on the beam only when the shutter opens. This happens automatically for some microscopes, but for those that do not have a pre-specimen shutter, you must do it yourself. Good luck. Yours, Bill Tivol, PhD EM Scientist Electron Cryo-Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
==============================Original Headers============================== 5, 24 -- From tivol-at-caltech.edu Thu May 3 19:50:17 2007 5, 24 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 5, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l440oGf5008238 5, 24 -- for {microscopy-at-msa.microscopy.com} ; Thu, 3 May 2007 19:50:17 -0500 5, 24 -- Received: from fire-dog.its.caltech.edu (fire-dog [192.168.1.4]) 5, 24 -- by water-ox-postvirus (Postfix) with ESMTP id 6A9BF2F053 5, 24 -- for {microscopy-at-msa.microscopy.com} ; Thu, 3 May 2007 17:50:16 -0700 (PDT) 5, 24 -- Received: from [192.168.159.233] (jpix-01.caltech.edu [131.215.2.133]) 5, 24 -- (using TLSv1 with cipher RC4-SHA (128/128 bits)) 5, 24 -- (No client certificate requested) 5, 24 -- by water-ox.its.caltech.edu (Postfix) with ESMTP id 232AE2EF35 5, 24 -- for {microscopy-at-msa.microscopy.com} ; Thu, 3 May 2007 17:50:15 -0700 (PDT) 5, 24 -- Mime-Version: 1.0 (Apple Message framework v624) 5, 24 -- In-Reply-To: {200705032341.l43Nf5rO000684-at-ns.microscopy.com} 5, 24 -- References: {200705032341.l43Nf5rO000684-at-ns.microscopy.com} 5, 24 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 5, 24 -- Message-Id: {4c3c2afd531c61d5f23edf88a8d8bd74-at-caltech.edu} 5, 24 -- Content-Transfer-Encoding: 7bit 5, 24 -- From: Bill Tivol {tivol-at-caltech.edu} 5, 24 -- Subject: Re: [Microscopy] viaWWW: electron diffraction 5, 24 -- Date: Thu, 3 May 2007 18:01:36 -0700 5, 24 -- To: microscopy-at-msa.microscopy.com 5, 24 -- X-Mailer: Apple Mail (2.624) 5, 24 -- X-Spam-Scanned: at Caltech-ITS on fire-dog by amavisd-2.4.5 ==============================End of - Headers==============================
Just a few comments off hand without ever doing these types of samples.
Your problem will be similar to what happened to me with glass samples. You are putting too much energy into your sample and it is heating. There are several things you can do to help. You have to do things quickly, i.e. low dosage. You have to lower the amount of energy you are putting into the sample. The best way to do this is going to higher accelerating energy. Remember, diffraction is elastic and you don't loose energy in the sample via that route. If your energy is lower, you have more inelastic scattering and you are dumping energy into the sample with those types of scattering events. You probably are working on a 100 or 120 kV machine, but you didn't say so. And another thing that you can do is to cool your sample and use a liquid nitrogen stage.
Having said this, I don't have any idea what the higher energy will do to your sample in terms of radiation damage.
-Scott
Scott D. Walck, Ph.D. Technical Director South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
US Toll Free: 1-800-728-2233 Tel: (949) 492-2600 Fax: (949) 492-1499
www.southbaytech.com walck-at-southbaytech.com
-----Original Message----- X-from: jvtaylo-at-emory.edu [mailto:jvtaylo-at-emory.edu] Sent: Thursday, May 03, 2007 4:45 PM To: Walck-at-SouthBayTech.com
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying
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Email: jvtaylo-at-emory.edu Name: Jeannette Taylor
Organization: IM&MF / Emory University
Title-Subject: [Filtered] electron diffraction
Question: We are beginning to explore the use of electron diffraction in the study of crystalized biological molucules. We don't have the background here. Some of these molecules are very fragile and disappear before a pattern can be captured on film. An electron diffractin pattern is seen, briefly, then it disappears. We have been following what the micropscope manual instructs on generating electron diffractions and have gotten some advice from a metallurgist. But we need some more help. Using purchased standards, we have gotten some very nice diffraction patterns.
I have a conceptual problem with my TEM (tecnai G20 (twin)). I asked help to 2 engineers from FEI, they gave me opposite answers (!!). Here is my problem: The objective aperture is situated just under the specimen itself. So it should have no influence on the intensity of the beam hitting my sample.
In this case why does my formvar film dilate and sometimes blow up when I forget to insert the objective aperture but is extremely stabile when it is inserted?
Even more strange: the EDX detector is placed at an angle several degrees ABOVE the specimen. I cannot detect anything when the objective aperture is inserted, I must remove it to be able to read something!
I have the feeling that actually the obj aperture is at the same height as the specimen and not under it, but one engineer told me it was just under the specimen. Also, I dont understand why the formvar is sensible to the presence of the obj aperture if it is at the same height as my specimen.
Stephane
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
==============================Original Headers============================== 8, 19 -- From nizets2-at-yahoo.com Fri May 4 04:07:18 2007 8, 19 -- Received: from web37401.mail.mud.yahoo.com (web37401.mail.mud.yahoo.com [209.191.91.133]) 8, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l4497IDu015715 8, 19 -- for {microscopy-at-microscopy.com} ; Fri, 4 May 2007 04:07:18 -0500 8, 19 -- Received: (qmail 1331 invoked by uid 60001); 4 May 2007 09:07:17 -0000 8, 19 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 8, 19 -- s=s1024; d=yahoo.com; 8, 19 -- h=X-YMail-OSG:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding:Message-ID; 8, 19 -- b=cxjvojOt1hILZ5FpG/RJjwZiYR+TRuPhGte0R1gRJYs2GN9EKdNiPEk7QGluBxPxIvAH1aKPdZ1qyQOJaZ+VMWsMYPMy42xiJUV6JaWUASOtb7jECnEm2duqCfucQ4k7fo7YZ8Sx4dKfbOKOSDa9rKQKY0sO11U6F8tPfy+A34Y=; 8, 19 -- X-YMail-OSG: WZe6wQIVM1nH8ooo4KNQBfPzy6R8e_bg9GZt0lWRlZR6EDVCIYMGPIJ07FlCJC1PXrt4kBteVPUoSlVypKhFxkHtDxp7tFLkL6xVBNurfNUjXRaMKM3Vex7IRE6fWw-- 8, 19 -- Received: from [80.122.101.102] by web37401.mail.mud.yahoo.com via HTTP; Fri, 04 May 2007 02:07:17 PDT 8, 19 -- Date: Fri, 4 May 2007 02:07:17 -0700 (PDT) 8, 19 -- From: Stephane Nizet {nizets2-at-yahoo.com} 8, 19 -- Subject: TEM : objective aperture and EDX 8, 19 -- To: microscopy-at-microscopy.com 8, 19 -- MIME-Version: 1.0 8, 19 -- Content-Type: text/plain; charset=iso-8859-1 8, 19 -- Content-Transfer-Encoding: 8bit 8, 19 -- Message-ID: {399888.608.qm-at-web37401.mail.mud.yahoo.com} ==============================End of - Headers==============================
Sorry but I had missed the 'Ask-A-Miicroscpist' on the header and assumed you were on the list-server (so I didn't email you my reply). I attach my original musings on the subject of vibration feedback and optical microscopes below, i.e. the microscope rooms don't really have to have special modifications at all for light microscopy provided stout worktops and/or standard microscope anti-vibrational systems are used as supplied my microscope manufacturers and their support industries.
It's only high magnification electron microscopes (EM) that go to 20,000x magnification that might benefit from more advanced modification of the room itself for anti-vibration (and these EM devices are too big and heavy to be used with light microscope anti-vibration systems) - but I'm mainly a light microscopist these days and not really the person the ask about EMs, you need an EM microscopist for this (besides it's not in your brief).
In the old days optical microscope support worktops were made of heavy damping materials like granite and slate and these work well, but you still need some form of rubber decoupling material as well, e.g. another heavy plate supporting the microscope and an anti-vibrational rubber type material underneath like squash balls or stacks of Fabreeka FabCel sheets to isolate the microscope (plus the little £5 sticky rubber feet on the microscope help quite a bit). Modern laminated worktops are fine provided they are 5cm+ thickness and well supported - see photos cheap £500 (bad) and micro-dissection (much better).
For expensive confocal or live cell time-lapse microscopes the best option is often a stand-alone air-table (powered by compressed air from a £500 electric pump). These cost around £5,000 for a standard design, which in comparison to the £200,000 for the confocal microscope isn't that much. The air table needs no special room modification, just a space for it and its £500 air pump. It's a standalone device that can move with the microscope to another room is needed. For a simple £10,000 laboratory upright microscope used by students for fixed slides the sticky rubber feet on the microscope are generally enough (a granite/slate support worktop might be nice though, but a standard thick heavy laminated wooden 'woodchip' one will be fine).
My comments on the rooms air conditioning, below, are only critically important if you wish to do time-lapse studies on the movement of living cells (where focus and XY stage drift totally ruin the time-lapse video). It can also affect a motorised automatic scan of a slide. However if you are manually looking at slides or fixed samples under the microscope variation in room temperature is less important, as you will simply refocus the optics or move the stage when it slowly drifts - if you notice it at all (the thermal changes being quite slow). You will still want the temperature to be comfortable for long periods of sitting though (about 22oC), and the worktop should be right height for comfortable microscope viewing with a fully adjustable chair.
A few examples of Vibration isolation Air-tables http://www.technicalmanufacturing.com/portals/lifescience.html http://www.kineticsystems.com/page135.html http://www.speirsrobertson.com/ http://www.nextdayscience.com/store/laboratory/anti-vibration-tables.html
Rubber isolation examples http://www.fabreeka.com/products/fabcel_pads.htm Some Fabcel can be seen in cheap £500 (brown in this case, but the black looks nicer) plus there are squashballs, anti-vibration rubber feet etc...there are loads of different types about see http://www.rswww.com and search anti vibration.
So generally you select the type of anti-vibrational microscope support you need, allowing for cost and whether you need high magnification for live cell work (the culture media and living cells wobble far more than fixed slides and need better support - fixed slides & microscopes tend to wobble together in unison so you don't notice it so much). Other than stout worktops most anti-vibrational devices are added after the room is built and so can be moved to another room with the microscope. In all cases our microscopes are used in standard laboratories with concrete floors although they have the room to themselves.
A few examples of the anti-vibration tables/plates we use for microscopes are attached. Note the very flimsy worktop under the cheap £500 black in-house manufactured damping plate, and to be honest we only needed the damping isolation plate with this microscope for our live cell work, not for fixed slides - the microscope had been used for 5 years previously just sitting on the worktop. The laser dissection microscope needs high magnifications to uv laser cut out individual cells and even chromosomes from fixed slides, but this was supplied by Zeiss with a simple steel plate and rubber feet for 'anti-vibration' which you can see resting on the black worktop (a better worktop than in the 'cheap £500' photo). This is all Ziess/PALM recommend for installation (and it works fine).
My original list-server message is attached below
Hope this is all OK, you probably know a lot of this anyway but any queries and just ask. Sorry you didn't get this earlier.
Keith
PS. Only Sandra gets the photo's I'm afraid (four photos showing a Leica SP2 confocal with a £3,000 Leica granite/squashball isolation table, a Bio-Rad [Zeiss Microscience] Radiance confocal with a £5,000 air table, a PALM/Zeiss Axiovert 200 micro-dissection system with PALM worktop damping isolation plate, and our cheap £500 Fabcel/140kg plate used with a Zeiss Axiovision 100M/OpenLabs time-lapse system.
--------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Institute of Ophthalmology UCL, London EC1V 9EL
-----Original Message----- X-from: Keith Morris [mailto:keith.morris-at-ucl.ac.uk] Sent: 02 May 2007 09:50 To: 'microscopy-at-microscopy.com'
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sandra.filippi-at-montgomerycollege.edu) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, April 30, 2007 at 20:12:50 ---------------------------------------------------------------------------
Email: sandra.filippi-at-montgomerycollege.edu Name: Sandra Lee Filippi, Campus Planner
Organization: montgomery college
Education: Graduate College
Location: Rockville MD
Question:
Dear Sir or Madam Microscopist:
Montgomery College is in design development for a new academic teaching science center. The lab planner for the design team is citing ASHRAE TC2.6 Guideline Criteria for use in Planning New Facilities and insists that we design to the VC-B level because the biology department uses compound microscopes with total magnification of 1,000x (oil immersion). I managed academic science labs for a two-year college for more than 20 years. We routinely used light microscopes at total magnification of 1,000x (oil immersion) in a facility built in 1965 without special vibration control. When we opened our new science center in 1995 it did not have special vibration control either. I have studied at Georgetown University and did student research at The NIH. Neither of these facilities required special vibration control for a light microscope. I have consulted with the Westover Scientific Customer Service/Product Specialist ñ Microscopy who states, ìTo be honest with you I have never heard of such a thing, neither have my colleagues. ÝThere are tables that are manufactured to reduce vibrations, but Iíve never heard of a building being designed around a microscope. ÝWe have customers who use microscopes in old buildings all over the world. Westover recommends that a microscope be used on a vibration free surface which typically means not locating your microscope on a table with other equipment that can cause vibrations such as fans and other lab equipment. Microscopes have been used for years and years on standard benchtops and other tables without issue. It's not our position to recommend additional support when building a new structure. In my entire career I have never heard of this requirement.î Westover manufactures microscopes for Fisher Scientific. The lab planner is going to cost us dearly unless I can refute his recommendation. What is your professional opinion in this matter? Thank you very much for your time. Sandra
Sandra Lee Filippi, Campus Planner Montgomery College Suite 200 40 West Gude Drive Room 223 Rockville MD 20850-1166 301.251.7362 vox 301.251.7379 fax 240.882.6672 cell
You have a very common problem. Firstly the objective aperture sits below the specimen, usually in the back focal plane of the objective. The films become damaged when you remove the aperture as the presence of the aperture acts to neutralise the charge on the film. My guess is when you have the objective aperture in place the x-ray signal is so great it swamps the detector with x-rays causing the detector to go 100% dead.
Try using very small spot sizes (condenser 1 at a higher strength), or smaller condenser apertures, or a lower emission current from the gun, all of which will help preserve the specimen. If you still have a problem try two of the above or even all three. The object here is to reduce the number of electrons hitting the specimen thus reducing the damaging component. If you still have a problem with film damage then use the largest objective aperture that you can find, even replacing one aperture with one of the smaller apertures used in the condenser system. I have to say that I am not sure if your microscope uses disk apertures or an aperture strip? The idea being that if you still need the objective aperture for stability the bigger the hole the lower the chance of picking up its x-rays.
I hope this helps?
Steve Chapman Senior Consultant Protrain For electron microscopy consultancy and training world wide Tel +44 1280 816512 Fax +44 1280 814007 Mobile +44 7711 606967 Web www.emcourses.com
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Email: wvrenter-at-sckcen.be Name: Wouter Van Renterghem
Organization: SCK-CEN
Title-Subject: [Filtered] orientation of the diffraction pattern
Question: When you defocus a diffraction pattern, you can see a small image of the selected area in the 000 spot. When you go from underfocus to overfocus the image of the selected area is rotated over 180ƒ. Can somebody tell me which of the two small images has the same orientation as the image in image mode? Do I have to underfocus the diffraction pattern or overfocus it?
Here is the May 2007 Microscopy Today table of contents. I will close the subscription list for this issue on Wednesday, May. 9th, 2007.
Microscopists in North America and MSA members anywhere can subscribe for free. Anyone else may subscribe for US$50 per year (to PARTIALLY cover postage). All subscriptions at http://www.microscopy-today.com Thank you.
Ron Anderson, Editor ================================ Illuminating the Formation of Lumens Stephen W. Carmichael, Mayo Clinic, Rochester, MN
Moore’s Law: A Review of Feature Size Shrinkage and its Effect on Microscopy in the Semiconductor Industry John Mardinly, Intel Corporation, Santa Clara, CA
An SEM Analysis Of Amphipleura pellucida with New Findings Gary Gaugler, Microtechnics, Inc., Granite Bay, CA
The Proposed Doppler Electron Velocimeter and the Need for Nanoscale Dynamics Phillip L. Reu, Sandia National Lab., Albuquerque, NM
Resolution and Sampling in Digital Imaging Ian Dobbie, National University of Ireland, Galway, Ireland
Use of Astronomy Filters in Light Microscopy and Photomicrography Jörg Piper, Clinic Meduna, Bad Bertrich, Germany
Analyzing the Growth of Magmatic Crystals – An Electron Microprobe Analysis Study Robert Sturm, University of Salzburg, Austria
µManager: Open Source Software for Light Microscope Imaging Nico Stuurman, Nenad Amdodaj and Ron Vale, University of California, San Francisco, CA
Vapor Coating: A Simple, Economical Procedure for Preparing Difficult Specimens for Scanning Electron Microscopy E. Ann Ellis and Michael W. Pendleton, Texas A&M University, College Station, TX
Students and the SEM: The First Encounter V.M.Dusevich, J.D.Eick, Univ. of Missouri, Kansas City, MO
The Magnification Myth Sander Stoks and Bas Groen
Quality Controls on SEM Performance: A Novel Reference Sample Brendan J. Griffin and Sharon T. Platten, Univ. of Western Australia, Crawley, Australia
Industry News
NetNotes SPECIMEN PREPARATION - LR White polymerization SPECIMEN PREPARATION - pollen grains SPECIMEN PREPARATION - tripod polishing problems SPECIMEN PREPARATION - ruthenium tetroxide staining recipe SPECIMEN PREPARATION – embedding mouse lens & eye SPECIMEN PREPARATION - pre-embedding cells in agar SPECIMEN PREPARATION - annealing tantalum IMMUNOCTYOCHEMISTRY – BSA purity for use as blocking agent IMAGE ANALYSIS – phase analysis PHOTOGRAPHY – neutral density filters DARKROOM - uneven printing CAMERAS - CMOS versus CCD MICROSCOPY - analysis of paper LM - Koehler illumination LM - calibration standard Z direction EM - field sources TEM - cleaning Pt apertures TEM - free lens control SEM - takeoff angle SEM - shorted lead wires - current contrast? SEM - lines on slow scan/capture Electron beam simulation
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HI, Tom- In our experience, this seems a probable resultdue to re-oxidation of the osmium in the dehydrating solutions. We never figured out what the oxidant is in the ethanol, but we solved the problem by using hexylene glycol as a dehydrating fluid in place of ethanol. You can find a protocol on our web site. Just follow the procedure in the Transmission Electron Microscopy section. Carol
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Email: cantonpa-at-unive.it Name: Patrizia Canton
Organization: University of Venice
Title-Subject: [Filtered] LaB6 filament
Question: We had a new LaB6 filament (Denka, M3,LKSH, tip shape 60†, 10micron mR Pointed) installed in our Jeol 3010. Jeol engineer had some problems during the installation because there were some black stripes visible in the filament image. He tried to change te distance filament-Wehnelt three times, to work with bias adjustment, with filament saturation, after 5 days he had to end his job because we finished the budget but with no good results since he only obtained emission current of 1 microA, and low current density. Now we are struggling with this filament and coming to nowhere. Is there anyone that can give some hints, suggestions, bibliography? Thanks Patrizia
Dear Stephane, Yes, the objective aperture sits just below the specimen and it is a well-known phenomenon that Formvar film will "pop" if you observe the specimen without inserting the objective aperture. The way it was explained to me was that, when you look at your sample without the objective aperture, all the radiation comes from one side and the charging, uneven heating and radiation causes thermal stress and deformation that causes the film to bulge and then, sometimes, break. When you insert the objective aperture, the radiation reflects from the aperture and balances the heat and radiation from the top, so there is not the stress of irradiating only one side. For the EDX, remember that the specimen is essentially transparent to the electron beam and the x-rays produced. The enormous flux of x-rays and back-scattered electrons generated from the beam hitting the solid metal of the aperture metal with 120kV electrons floods the EDX detector and "kills" it, basically generating 100% dead time. It may take several minutes to recover, after you remove the aperture. My sequence to switch to EDX analysis always includes removing the objective aperture first. I find doing EDX analysis on the TEM to be a careful balance between zero x-rays in thin films and 100% dead time when you hit something too thick, like a grid bar. They look the same on the EDX display. I am mainly working in Materials Engineering, so I do lots of EDX on my TEM, but I don't use Formvar films. I use carbon evaporated films that I make myself and they seem to withstand 200kV TEM examination with or without an objective aperture inserted and they are already conductive. Good luck. Regards,
-----Original Message----- X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] Sent: May 4, 2007 2:17 AM To: mager-at-interchange.ubc.ca
Deal all!
I have a conceptual problem with my TEM (tecnai G20 (twin)). I asked help to 2 engineers from FEI, they gave me opposite answers (!!). Here is my problem: The objective aperture is situated just under the specimen itself. So it should have no influence on the intensity of the beam hitting my sample.
In this case why does my formvar film dilate and sometimes blow up when I forget to insert the objective aperture but is extremely stabile when it is inserted?
Even more strange: the EDX detector is placed at an angle several degrees ABOVE the specimen. I cannot detect anything when the objective aperture is inserted, I must remove it to be able to read something!
I have the feeling that actually the obj aperture is at the same height as the specimen and not under it, but one engineer told me it was just under the specimen. Also, I dont understand why the formvar is sensible to the presence of the obj aperture if it is at the same height as my specimen.
Stephane
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==============================Original Headers============================== 16, 35 -- From mager-at-interchange.ubc.ca Fri May 4 11:30:32 2007 16, 35 -- Received: from mr8.mail-relay.ubc.ca (mr8.mail-relay.ubc.ca [137.82.45.15]) 16, 35 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l44GUWvc029197 16, 35 -- for {microscopy-at-microscopy.com} ; Fri, 4 May 2007 11:30:32 -0500 16, 35 -- Received: from mr8.mail-relay.ubc.ca (localhost [127.0.0.1]) 16, 35 -- by localhost (Postfix) with SMTP id 842C310A30 16, 35 -- for {microscopy-at-microscopy.com} ; Fri, 4 May 2007 09:30:31 -0700 (PDT) 16, 35 -- Received: from mta1.interchange.ubc.ca (mta1.interchange.ubc.ca [142.103.145.69]) 16, 35 -- by mr8.mail-relay.ubc.ca (Postfix) with ESMTP 16, 35 -- for {microscopy-at-microscopy.com} ; Fri, 4 May 2007 09:30:30 -0700 (PDT) 16, 35 -- Received: from Mary (desk.staff.mmat.ubc.ca [137.82.16.178]) 16, 35 -- by smtp.interchange.ubc.ca 16, 35 -- (iPlanet Messaging Server 5.2 HotFix 1.21 (built Sep 8 2003)) 16, 35 -- with ESMTPS id {0JHI008H7Z6UJO-at-smtp.interchange.ubc.ca} for 16, 35 -- microscopy-at-microscopy.com; Fri, 04 May 2007 09:30:30 -0700 (PDT) 16, 35 -- Date: Fri, 04 May 2007 09:30:08 -0700 16, 35 -- From: Mary Mager {mager-at-interchange.ubc.ca} 16, 35 -- Subject: RE: [Microscopy] TEM : objective aperture and EDX 16, 35 -- In-reply-to: {200705040916.l449GX72024583-at-ns.microscopy.com} 16, 35 -- To: nizets2-at-yahoo.com 16, 35 -- Cc: "'Microscopy'" {microscopy-at-microscopy.com} 16, 35 -- Reply-to: mager-at-interchange.ubc.ca 16, 35 -- Message-id: {0JHI008H8Z6UJO-at-smtp.interchange.ubc.ca} 16, 35 -- Organization: Materials Eng. UBC 16, 35 -- MIME-version: 1.0 16, 35 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 16, 35 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 16, 35 -- Content-type: text/plain; charset=US-ASCII 16, 35 -- Content-transfer-encoding: 7bit 16, 35 -- Thread-index: AceOLOjfjQBeD3PHQeO+5qgq3sXG2QAOenow 16, 35 -- X-UBC-Scanned: Sophos PureMessage 5.3.1.294258, Antispam-Engine: 2.5.0.283055, Antispam-Data: 2007.5.4.91133 16, 35 -- X-UBC-Relayed: Relayed through mail-relay.ubc.ca 16, 35 -- X-PerlMx-Spam: Probability=7%, Report=__C230066_P2 0, __C230066_P5 0, __CP_URI_IN_BODY 0, __CT 0, __CTE 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __HAS_X_MAILER 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0, __STOCK_PHRASE_24 0, __STOCK_PHRASE_7 0, __USER_AGENT_MS_GENERIC 0 16, 35 -- X-Spam-Level: 16, 35 -- X-Spam-Flag: No ==============================End of - Headers==============================
On May 3, 2007, at 7:07 PM, walck-at-southbaytech.com wrote:
} Just a few comments off hand without ever doing these types of samples. } } Your problem will be similar to what happened to me with glass } samples. You } are putting too much energy into your sample and it is heating. There } are } several things you can do to help. You have to do things quickly, } i.e. low } dosage. You have to lower the amount of energy you are putting into } the } sample. The best way to do this is going to higher accelerating } energy. } Remember, diffraction is elastic and you don't loose energy in the } sample } via that route. If your energy is lower, you have more inelastic } scattering } and you are dumping energy into the sample with those types of } scattering } events. You probably are working on a 100 or 120 kV machine, but you } didn't } say so. And another thing that you can do is to cool your sample and } use a } liquid nitrogen stage. } } Having said this, I don't have any idea what the higher energy will do } to } your sample in terms of radiation damage. } Dear Scott, Having done considerable ED at voltages from 100 to 1200 kV, I'd like to offer a few minor corrections to your post. You are correct that the problem is that too much energy was being deposited, but heating is not the effect that causes the pattern to decay. Changes in the specimen that destroy the crystalline order are responsible, and these are caused by breaking of chemical bonds, ionization, and other processes, such as displacement of atoms. Going to a higher energy does give advantages for ED, principally making the scattering closer to kinematic and getting higher resolution spots due to the flatter Ewald sphere. As the energy of the beam increases, the total scattering cross-section decreases, which results in less energy deposited, but the ratio of elastic scattering to inelastic scattering also decreases, so the damage-to-information ratio increases, but this effect is minor, and excellent ED patterns can be obtained at 1200 kV. Yours, Bill Tivol, PhD EM Scientist Electron Cryo-Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
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Hi all, I am looking for a carbon rod sharpened for rods with diameter of 0.12 inch diameter rods. We had a great electrical
one that rotated the rod and you just moved a small cutting edge along a track to cut a nice long and narrow rod at the end of the larger rod. The rotating shaft no longer is stable so rods break too easily.
Would appreciate advice as to replacements and would prefer electrical one rather than the hand-held type.
Debby
Debby Sherman, Director Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www.agriculture.purdue.edu/microscopy
==============================Original Headers============================== 6, 21 -- From dsherman-at-purdue.edu Fri May 4 14:22:16 2007 6, 21 -- Received: from 1061exfe04.adpc.purdue.edu (1061exfe04.adpc.purdue.edu [128.210.63.227]) 6, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l44JMG3S031857 6, 21 -- for {microscopy-at-microscopy.com} ; Fri, 4 May 2007 14:22:16 -0500 6, 21 -- Received: from exchange.purdue.edu ([172.21.6.24]) by 1061exfe04.adpc.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 6, 21 -- Fri, 4 May 2007 15:22:16 -0400 6, 21 -- Received: from 128.210.161.94 ([128.210.161.94]) by EXCH04.purdue.lcl ([172.21.6.26]) via Exchange Front-End Server exch.itap.purdue.edu ([128.210.63.104]) with Microsoft Exchange Server HTTP-DAV ; 6, 21 -- Fri, 4 May 2007 19:22:16 +0000 6, 21 -- User-Agent: Microsoft-Entourage/11.3.3.061214 6, 21 -- Date: Fri, 04 May 2007 15:22:15 -0400 6, 21 -- Subject: Carbon rod sharpener 6, 21 -- From: Debby Sherman {dsherman-at-purdue.edu} 6, 21 -- To: "message to: MSA list" {microscopy-at-microscopy.com} 6, 21 -- Message-ID: {C2610027.1BBEB%dsherman-at-purdue.edu} 6, 21 -- Thread-Topic: Carbon rod sharpener 6, 21 -- Thread-Index: AceOgYXLxEBWnvp0EduF9AARJN08Mg== 6, 21 -- Mime-version: 1.0 6, 21 -- Content-type: text/plain; 6, 21 -- charset="US-ASCII" 6, 21 -- Content-transfer-encoding: 7bit 6, 21 -- X-OriginalArrivalTime: 04 May 2007 19:22:16.0250 (UTC) FILETIME=[868A89A0:01C78E81] ==============================End of - Headers==============================
Hi Debby- the one I have is about a gazillion years old (did they do EM then??) but it still works beautifully. It was from Ladd. I don't know if they still sell them, you'd have to check their online catalog. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Weill-Cornell Medical College
Check out our's at http://www.laddresearch.com/Key_Products/Digital_High_Vacuum_Evaporator/VacEvap/Carbon_Rod_Sharpening/carbon_rod_sharpening.html It sounds like what you are looking for.
Disclaimer: Ladd Research sells carbon rod sharpeners, carbon rods, carbon rod points, etc.
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==============================Original Headers============================== 19, 27 -- From jd-at-laddresearch.com Fri May 4 14:41:52 2007 19, 27 -- Received: from lancia.electric.net (lancia.electric.net [216.129.90.248]) 19, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l44Jfq4Y018616 19, 27 -- for {microscopy-at-microscopy.com} ; Fri, 4 May 2007 14:41:52 -0500 19, 27 -- Received: from root by lancia.electric.net with emc1-ok (Exim 4.62) 19, 27 -- (envelope-from {jd-at-laddresearch.com} ) 19, 27 -- id 1Hk3f5-0005Ue-Td; Fri, 04 May 2007 12:41:51 -0700 19, 27 -- Received: by emcmailer; Fri, 04 May 2007 12:41:51 -0700 19, 27 -- Received: from [216.204.198.170] (helo=NewServer.laddresearch.com) 19, 27 -- by lancia.electric.net with esmtps (TLSv1:AES256-SHA:256) 19, 27 -- (Exim 4.62) 19, 27 -- (envelope-from {jd-at-laddresearch.com} ) 19, 27 -- id 1Hk3f4-0005TQ-U6; Fri, 04 May 2007 12:41:50 -0700 19, 27 -- X-Mailer: QUALCOMM Windows Eudora Version 7.1.0.9 19, 27 -- Date: Fri, 04 May 2007 15:41:20 -0400 19, 27 -- To: dsherman-at-purdue.edu 19, 27 -- From: jd {jd-at-laddresearch.com} 19, 27 -- Subject: Re: [Microscopy] Carbon rod sharpener 19, 27 -- Cc: Microscopy listserver {microscopy-at-microscopy.com} 19, 27 -- In-Reply-To: {200705041929.l44JTE4F010128-at-ns.microscopy.com} 19, 27 -- References: {200705041929.l44JTE4F010128-at-ns.microscopy.com} 19, 27 -- Mime-Version: 1.0 19, 27 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 19, 27 -- X-Outbound-IP: 216.204.198.170 19, 27 -- X-Env-From: jd-at-laddresearch.com 19, 27 -- X-Virus-Status: Scanned by VirusSMART (c) 19, 27 -- Message-Id: {E1Hk3f5-0005Ue-Td-at-lancia.electric.net} ==============================End of - Headers==============================
I agree that electrons coming from the objective aperture balance the effect on the formvar film caused by the incident beam, as pointed out by Dr Steve Chapman and Dr Mary Fletcher. However, I doubt the force that breaks the film. The energy of the incident electrons is of the order 100keV. The inelastic cross-section for this energy loss is extremely small. So it is not likely there are a lot incident electrons are stopped and accumulate on the film. Heat will certainly be produced under illumination. But inelastic scattering can happen at any depth, especially in a very thin film where most events are single-scattering. So the heat should not differ too much between the two surface. I have a bold proposal. The film is broken by repulsive electrostatic force. But the charge is possitive instead of negative. The formvar mainly contains C, O, N and H which are light elements and easy to ionize. So some atoms in the film lose valence electrons and become possitively charged. Dey and Williams (J. Phys. D, 1988, vol.21, 108) studied the EELS spectra of formvar film. They observed low binding energy peaks at 11-93eV. Scatterings in this range have considerable cross-sections and can involve many atoms. On the other hand, the electrons coming from the objective aperture, either back scattered or secondary electrons, have lower energy compared to the incident beam and, hence easier to be trapped by the film. So these electrons neutrolize the possitive charge and stabilize the film. This is just my personal speculation after reading posts on this topic. Please feel free to correct me.
Shu Miao
On Fri, 4 May 2007 mager-at-interchange.ubc.ca wrote:
} } Dear Stephane, } Yes, the objective aperture sits just below the specimen and it is a } well-known phenomenon that Formvar film will "pop" if you observe the } specimen without inserting the objective aperture. The way it was explained } to me was that, when you look at your sample without the objective aperture, } all the radiation comes from one side and the charging, uneven heating and } radiation causes thermal stress and deformation that causes the film to } bulge and then, sometimes, break. When you insert the objective aperture, } the radiation reflects from the aperture and balances the heat and radiation } from the top, so there is not the stress of irradiating only one side. } For the EDX, remember that the specimen is essentially transparent to the } electron beam and the x-rays produced. The enormous flux of x-rays and } back-scattered electrons generated from the beam hitting the solid metal of } the aperture metal with 120kV electrons floods the EDX detector and "kills" } it, basically generating 100% dead time. It may take several minutes to } recover, after you remove the aperture. My sequence to switch to EDX } analysis always includes removing the objective aperture first. I find doing } EDX analysis on the TEM to be a careful balance between zero x-rays in thin } films and 100% dead time when you hit something too thick, like a grid bar. } They look the same on the EDX display. } I am mainly working in Materials Engineering, so I do lots of EDX on my TEM, } but I don't use Formvar films. I use carbon evaporated films that I make } myself and they seem to withstand 200kV TEM examination with or without an } objective aperture inserted and they are already conductive. } Good luck. } Regards, } } Mary Fletcher (nee Mager) } Electron Microscopist } Materials Eng. UBC } #309 - 6350 Stores Road } Vancouver, B.C. V6T 1Z4 } Canada } Tel: 604-822-5648 } Fax: 604-822-3619 } email: mager-at-interchange.ubc.ca } } -----Original Message----- } } } Deal all! } } I have a conceptual problem with my TEM (tecnai G20 } (twin)). I asked help to 2 engineers from FEI, they } gave me opposite answers (!!). } Here is my problem: } The objective aperture is situated just under the } specimen itself. So it should have no influence on the } intensity of the beam hitting my sample. } } In this case why does my formvar film dilate and } sometimes blow up when I forget to insert the } objective aperture but is extremely stabile when it is } inserted? } } Even more strange: the EDX detector is placed at an } angle several degrees ABOVE the specimen. I cannot } detect anything when the objective aperture is } inserted, I must remove it to be able to read } something! } } I have the feeling that actually the obj aperture is } at the same height as the specimen and not under it, } but one engineer told me it was just under the } specimen. Also, I dont understand why the formvar is } sensible to the presence of the obj aperture if it is } at the same height as my specimen. } } } Stephane
==============================Original Headers============================== 5, 21 -- From shu-at-caltech.edu Fri May 4 15:18:36 2007 5, 21 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 5, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l44KIarf002326 5, 21 -- for {Microscopy-at-microscopy.com} ; Fri, 4 May 2007 15:18:36 -0500 5, 21 -- Received: from earth-dog.its.caltech.edu (earth-dog [192.168.1.3]) 5, 21 -- by fire-ox-postvirus (Postfix) with ESMTP id 0E368352BA 5, 21 -- for {Microscopy-at-microscopy.com} ; Fri, 4 May 2007 13:18:36 -0700 (PDT) 5, 21 -- Received: from blinky (blinky.its.caltech.edu [131.215.239.33]) 5, 21 -- by earth-ox.its.caltech.edu (Postfix) with ESMTP id 006F636FE4 5, 21 -- for {Microscopy-at-microscopy.com} ; Fri, 4 May 2007 13:18:33 -0700 (PDT) 5, 21 -- Date: Fri, 4 May 2007 13:18:33 -0700 (PDT) 5, 21 -- From: Shu Miao {shu-at-caltech.edu} 5, 21 -- X-X-Sender: shu-at-blinky 5, 21 -- To: Microscopy-at-microscopy.com 5, 21 -- Subject: [Microscopy] RE: TEM : objective aperture and EDX 5, 21 -- In-Reply-To: {200705041635.l44GZ2DA008573-at-ns.microscopy.com} 5, 21 -- Message-ID: {Pine.GSO.4.58.0705041055390.7185-at-blinky} 5, 21 -- References: {200705041635.l44GZ2DA008573-at-ns.microscopy.com} 5, 21 -- MIME-Version: 1.0 5, 21 -- Content-Type: TEXT/PLAIN; charset=US-ASCII 5, 21 -- X-Spam-Scanned: at Caltech-ITS on earth-dog by amavisd-2.4.5 ==============================End of - Headers==============================
On May 4, 2007, at 9:28 AM, cantonpa-at-unive.it wrote:
} Question: We had a new LaB6 filament (Denka, } M3,LKSH, tip shape 60†, 10micron mR Pointed) } installed in our Jeol 3010. } Jeol engineer had some problems during the } installation because there were some black } stripes visible in the filament image. He tried } to change te distance filament-Wehnelt three } times, to work with bias adjustment, with } filament saturation, after 5 days he had to end } his job because we finished the budget but with } no good results since he only obtained emission } current of 1 microA, and low current density. Now } we are struggling with this filament and coming } to nowhere. Is there anyone that can give some } hints, suggestions, bibliography? } Dear Patrizia, As the current through a LaB6 filament is increased, the first part of the filament that emits electrons is the flat part of the tip, which is a truncated pyramid. The next parts of the filament to emit are the faces of the pyramid, and finally, the last parts to emit are the edges between the flat faces. If you adjust the condenser to crossover and put the mag to about 30 kx, so you can see the image of the filament, you will first see a very small, round spot as the tip starts to emit, then four larger, brighter spots will appear around the first one, these will get larger and brighter and fill out the area until only four dark streaks, where the emission from the edges would be, remain, and finally, these will also fill in giving a uniform spot when the filament is saturated. This assumes that the filament is properly centered in the Wehnelt aperture. The failure to achieve saturation and the low emission current sound like the filament is not getting enough current. If the black stripes were not evenly spaced and along radial directions in the filament image, then it is likely that the tip was damaged, and the damaged parts are not emitting. Yours, Bill Tivol, PhD EM Scientist Electron Cryo-Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
==============================Original Headers============================== 5, 23 -- From tivol-at-caltech.edu Fri May 4 15:47:36 2007 5, 23 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 5, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l44KlaPf014117 5, 23 -- for {microscopy-at-msa.microscopy.com} ; Fri, 4 May 2007 15:47:36 -0500 5, 23 -- Received: from earth-dog.its.caltech.edu (earth-dog [192.168.1.3]) 5, 23 -- by water-ox-postvirus (Postfix) with ESMTP id B0E212F0F8 5, 23 -- for {microscopy-at-msa.microscopy.com} ; Fri, 4 May 2007 13:47:35 -0700 (PDT) 5, 23 -- Received: from [192.168.159.233] (jpix-01.caltech.edu [131.215.2.133]) 5, 23 -- by wood-ox.its.caltech.edu (Postfix) with ESMTP id 85F982EEC6 5, 23 -- for {microscopy-at-msa.microscopy.com} ; Fri, 4 May 2007 13:47:34 -0700 (PDT) 5, 23 -- Mime-Version: 1.0 (Apple Message framework v624) 5, 23 -- In-Reply-To: {200705041628.l44GSdAE026752-at-ns.microscopy.com} 5, 23 -- References: {200705041628.l44GSdAE026752-at-ns.microscopy.com} 5, 23 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 5, 23 -- Message-Id: {ebcfd22b5068678775e269015f7c7e0d-at-caltech.edu} 5, 23 -- From: Bill Tivol {tivol-at-caltech.edu} 5, 23 -- Subject: Re: [Microscopy] [Filtered] MicroscopyListserverviaWWW: LaB6 filament 5, 23 -- Date: Fri, 4 May 2007 13:58:58 -0700 5, 23 -- To: microscopy-at-msa.microscopy.com 5, 23 -- X-Mailer: Apple Mail (2.624) 5, 23 -- X-Spam-Scanned: at Caltech-ITS on earth-dog by amavisd-2.4.5 5, 23 -- Content-Transfer-Encoding: 8bit 5, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l44KlaPf014117 ==============================End of - Headers==============================
On May 4, 2007, at 5:52 AM, wvrenter-at-sckcen.be wrote:
} Question: When you defocus a diffraction pattern, you can see a small } image of the selected area in the 000 spot. When you go from } underfocus to overfocus the image of the selected area is rotated over } 180ƒ. Can somebody tell me which of the two small images has the same } orientation as the image in image mode? Do I have to underfocus the } diffraction pattern or overfocus it? } Dear Wouter, The orientation of the pattern is not necessarily the same as that of the image in either under or over focus, and, furthermore, it can be different for different camera lengths. There should be information in the manual of your instrument that can tell you the difference in orientations. If not, put in a specimen with an asymmetric object, take an image in normal mode, then take one in defocussed diffraction mode and compare. An aggregate of gold beads on a carbon film is a good test object for this. Yours, Bill Tivol, PhD EM Scientist Electron Cryo-Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
==============================Original Headers============================== 5, 25 -- From tivol-at-caltech.edu Fri May 4 15:54:56 2007 5, 25 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 5, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l44Ksu9r025638 5, 25 -- for {microscopy-at-msa.microscopy.com} ; Fri, 4 May 2007 15:54:56 -0500 5, 25 -- Received: from earth-dog.its.caltech.edu (earth-dog [192.168.1.3]) 5, 25 -- by fire-ox-postvirus (Postfix) with ESMTP 5, 25 -- id 2FAE535291; Fri, 4 May 2007 13:54:52 -0700 (PDT) 5, 25 -- Received: from [192.168.159.233] (jpix-01.caltech.edu [131.215.2.133]) 5, 25 -- (using TLSv1 with cipher RC4-SHA (128/128 bits)) 5, 25 -- (No client certificate requested) 5, 25 -- by fire-ox.its.caltech.edu (Postfix) with ESMTP 5, 25 -- id 0A59435281; Fri, 4 May 2007 13:54:51 -0700 (PDT) 5, 25 -- Mime-Version: 1.0 (Apple Message framework v624) 5, 25 -- In-Reply-To: {200705041252.l44CqLr0021221-at-ns.microscopy.com} 5, 25 -- References: {200705041252.l44CqLr0021221-at-ns.microscopy.com} 5, 25 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 5, 25 -- Message-Id: {0aeb2542e401c93edde93ba34775b710-at-caltech.edu} 5, 25 -- From: Bill Tivol {tivol-at-caltech.edu} 5, 25 -- Subject: Re: [Microscopy] viaWWW: orientation of the diffraction 5, 25 -- Date: Fri, 4 May 2007 14:06:05 -0700 5, 25 -- To: wvrenter-at-sckcen.be, microscopy-at-msa.microscopy.com 5, 25 -- X-Mailer: Apple Mail (2.624) 5, 25 -- X-Spam-Scanned: at Caltech-ITS on earth-dog by amavisd-2.4.5 5, 25 -- Content-Transfer-Encoding: 8bit 5, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l44Ksu9r025638 ==============================End of - Headers==============================
For OsO4 to be reduced to OsO2 (as happens during fixation) requires the presence of hydrogen ions and an electron donor. Electrons can be supplied from breaking up the double bonds in fatty acid alkyl chains of unsaturated (membrane) lipids.
The reaction is reversible, so reoxidation is theoretically possible, but only if a stronger oxidizer with a redox potential more positive than the one associated with the OsO4/OsO2 redox couple (Eo =1.02 Volts) is present. And even then, re-oxidation will only occur under suitable circumstances! I found Carol's comment that hexylene glycol preserves the Osmium contrast very interesting. What could it be in ethanol that might re-oxidize the Osmium? Peroxides? Chlorine? Oxygen in statu nascendi? OsO4 as such is a pretty strong oxidizing agent already. Very puzzling.
I hope I am not making any serious mistakes here, I am not a chemist (I wish!), just trying to understand how it works, but it seems a slightly acidic environment would be good to promote the fixation as well as to preserve the OsO2 in the tissue after fixation. Could it be that the ethanol is alkaline? Even though ethanol is still somewhat polar, it may not take much in a non-aquaeous environment.
I am sure there will be proper chemists and physicists subsribing to this great listserver. Maybe they would be willing to look into our mostly empirically established procedures, we might be able to make a big move forward....
Jan Leunissen
Aurion - President Electron Microscopy Research Advisor Costerweg 5 Dept Anatomy and Structural Biology 6702 AA Wageningen Otago School of Medical Sciences The Netherlands Dunedin, New Zealand phone 31-317-497676 phone 64-3-4795465 fax 31-317-415955 fax 64-3-4797254 http://www.aurion.nl http://anatomy.otago.ac.nz -----------------------------------------------------------------
==============================Original Headers============================== 9, 19 -- From leunissen-at-aurion.nl Fri May 4 16:57:26 2007 9, 19 -- Received: from fep04.xtra.co.nz (fep04.xtra.co.nz [210.54.141.242]) 9, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l44LvPs9005396 9, 19 -- for {Microscopy-at-microscopy.com} ; Fri, 4 May 2007 16:57:26 -0500 9, 19 -- Received: from [192.168.1.50] (really [222.154.161.58]) by fep04.xtra.co.nz 9, 19 -- with ESMTP 9, 19 -- id {20070504215719.TYIZ25916.fep04.xtra.co.nz-at-[192.168.1.50]} 9, 19 -- for {Microscopy-at-microscopy.com} ; Sat, 5 May 2007 09:57:19 +1200 9, 19 -- Mime-Version: 1.0 (Apple Message framework v752.2) 9, 19 -- In-Reply-To: {200705041545.l44FjZZo014773-at-ns.microscopy.com} 9, 19 -- References: {200705041545.l44FjZZo014773-at-ns.microscopy.com} 9, 19 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 9, 19 -- Message-Id: {608CA1A0-F684-4BF7-A13E-130BE657893B-at-aurion.nl} 9, 19 -- Content-Transfer-Encoding: 7bit 9, 19 -- From: Jan Leunissen {leunissen-at-aurion.nl} 9, 19 -- Subject: Re: [Microscopy] Re: Help with enhancing contrast of Mitochondrial 9, 19 -- Date: Sat, 5 May 2007 09:57:18 +1200 9, 19 -- To: Microscopy-at-microscopy.com 9, 19 -- X-Mailer: Apple Mail (2.752.2) ==============================End of - Headers==============================
I'm completely dumbfounded by this one. I am in the process of checking for leaks on our SEM, and it pumps down perfectly one moment, then it won't even trigger PiG3. When I start to take things apart to check seals and such, then I put it back together (After finding nothing wrong) it evacuates perfectly. Then I let it sit overnight to see if there is a leak, and when I get in the next morning, it is completely at atmospheric pressure and then won't hold a vacuum and trigger PiG3 anymore. I'm a little confused at how it will hold a vacuum only the first time I pump it down after dismantling and reassembling parts of the vacuum system. I don't have a leak detector, but boy would I love one! (I also don't have any gauges capable of measuring absolute vacuum vs. relative vacuum, so I have no idea what the actual measurement in torr is.)
--Justin A. Kraft
==============================Original Headers============================== 2, 26 -- From kraftpiano-at-gmail.com Fri May 4 20:28:40 2007 2, 26 -- Received: from wr-out-0506.google.com (wr-out-0506.google.com [64.233.184.234]) 2, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l451Seww019490 2, 26 -- for {microscopy-at-microscopy.com} ; Fri, 4 May 2007 20:28:40 -0500 2, 26 -- Received: by wr-out-0506.google.com with SMTP id i31so1164508wra 2, 26 -- for {microscopy-at-microscopy.com} ; Fri, 04 May 2007 18:28:39 -0700 (PDT) 2, 26 -- DKIM-Signature: a=rsa-sha1; c=relaxed/relaxed; 2, 26 -- d=gmail.com; s=beta; 2, 26 -- h=domainkey-signature:received:received:message-id:date:from:to:subject:mime-version:content-type:content-transfer-encoding:content-disposition; 2, 26 -- b=cJmt3MZGjC95glyyFu+AdPFMqPSYoAxgMR+EEmmwxcGSmf9jAaZNhlAbKhDp8Lw0pkV58J8V4XUnfo2K8+l5Y4lPkodDfOgmAT8bDZwXxnh/Qw5RlVm+u2fQE07ZLgT4B1VLT6ZP9fhkPJG4MkB4qFE4yS/976P3LuVwAtAhZQ0= 2, 26 -- DomainKey-Signature: a=rsa-sha1; c=nofws; 2, 26 -- d=gmail.com; s=beta; 2, 26 -- h=received:message-id:date:from:to:subject:mime-version:content-type:content-transfer-encoding:content-disposition; 2, 26 -- b=j+NcTQR1YAWPfjXoMiGt0pCoHrffvabybEPYQGQALUh1uQocYfxunYiU3EnEnAAKmVjNCewAQ6SdNwXlbQMuD1iulfnngZH6DMhCKSh39cDDtCMhY/6cv+EVn4fQYR/uUglWyJ9NUB9r0JIk2UtZwVsOBnAxfzwvzLYLfnzir0s= 2, 26 -- Received: by 10.114.133.1 with SMTP id g1mr1349202wad.1178328519224; 2, 26 -- Fri, 04 May 2007 18:28:39 -0700 (PDT) 2, 26 -- Received: by 10.114.79.12 with HTTP; Fri, 4 May 2007 18:28:38 -0700 (PDT) 2, 26 -- Message-ID: {25e2b0d20705041828v19b4f286r2878534d132d125-at-mail.gmail.com} 2, 26 -- Date: Fri, 4 May 2007 21:28:38 -0400 2, 26 -- From: "Justin Kraft" {kraftpiano-at-gmail.com} 2, 26 -- To: microscopy-at-microscopy.com 2, 26 -- Subject: SEM: Intermittent Vacuum Leak 2, 26 -- MIME-Version: 1.0 2, 26 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 2, 26 -- Content-Transfer-Encoding: 7bit 2, 26 -- Content-Disposition: inline ==============================End of - Headers==============================
As soon as I saw Bruce, I thought of the Monty Python skit. Then there was our 3 year old daughter in the back seat (of othe car) one night as Mom & Dad (spouse and me) were discussing names for the impending baby (this was 1972, ok?) when she (in the back seat) piped up and said "What about Bruce?".
Roger Moretz, Ph.D.
On 5/3/07, frank.karl-at-degussa.com {frank.karl-at-degussa.com} wrote: } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Guilty! } } Having worn glasses since eight grade, I have come to associate optical } with glass the photons. } } Phototonic microscopy? sounds like a title of a paper in search of a } little primping } Non-modified bright field? - sounds like a title of a paper in search of a } little primping from a major university } Light microscopy? - sort of suggest their might be a heavy microscopy } Refractory microscopy - sound like microscopy in hell } Specular microscopy - the study of the stock market in great detail.. } } Tongue firmly implanted in cheek: Let's call it Bruce. I got the idea } from a microscopy society bog ( } http://www.msneo.org/2006/06/blog-mail.html) } who knows.......... } } Stay safe..............Frank } } } } } protrain-at-emcourse } s.com To: frank.karl-at-degussa.com } cc: } 05/03/2007 07:09 Subject: [Microscopy] Optical Microscopy } AM } Please respond to } protrain } } } } } } } } } ---------------------------------------------------------------------------- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } } Hi } } All the fuss over microprobes was fun but if you want to get your teeth } into } names used by scientists (?) what about "optical microscopy"? } } With my 43 years in the business I have always taught that whilst light } microscopes have "light optics" electron microscopes have "electron } optics", } the principles are the same. So when I see a piece about optical } microscopes I always look to see if it should be of interest to me? } } What is the general thought? } } Steve Chapman } Senior Consultant Protrain } For electron microscopy consultancy and training world wide } Tel +44 1280 816512 Fax +44 1280 814007 } Mobile +44 7711 606967 Web www.emcourses.com } } } ==============================Original } Headers============================== } 6, 26 -- From protrain-at-emcourses.com Thu May 3 06:06:37 2007 } 6, 26 -- Received: from smtp1.pri.skybb.uk.easynet.net } (smtp1.pri.skybb.uk.easynet.net [87.86.189.69]) } 6, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id l43B6bku028581 } 6, 26 -- for {microscopy-at-microscopy.com} ; Thu, 3 May 2007 } 06:06:37 -0500 } 6, 26 -- Received: from [90.200.252.11] (helo=advent) } 6, 26 -- by smtp1.pri.skybb.uk.easynet.net with smtp (Exim 4.62) } 6, 26 -- (envelope-from {protrain-at-emcourses.com} ) } 6, 26 -- id 1HjZ8u-0002y3-9V } 6, 26 -- for microscopy-at-microscopy.com; Thu, 03 May 2007 12:06:36 } +0100 } 6, 26 -- Message-ID: {001901c78d73$bc033230$0200a8c0-at-advent} } 6, 26 -- Reply-To: "Steve Chapman" {protrain-at-emcourses.com} } 6, 26 -- From: "Steve Chapman" {protrain-at-emcourses.com} } 6, 26 -- To: "American Soc" {microscopy-at-microscopy.com} } 6, 26 -- Subject: [Microscopy] Optical Microscopy } 6, 26 -- Date: Thu, 3 May 2007 12:10:57 +0100 } 6, 26 -- Organization: Protrain } 6, 26 -- MIME-Version: 1.0 } 6, 26 -- Content-Type: text/plain; } 6, 26 -- format=flowed; } 6, 26 -- charset="iso-8859-1"; } 6, 26 -- reply-type=original } 6, 26 -- Content-Transfer-Encoding: 7bit } 6, 26 -- X-Priority: 3 } 6, 26 -- X-MSMail-Priority: Normal } 6, 26 -- X-Mailer: Microsoft Outlook Express 6.00.2900.3028 } 6, 26 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 } ==============================End of - } Headers============================== } } } } } ==============================Original Headers============================== } 27, 18 -- From frank.karl-at-degussa.com Thu May 3 07:00:41 2007 } 27, 18 -- Received: from framailout1.rz.itson.com (mailout2.degussa.com [149.216.91.173]) } 27, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l43C0f20019657 } 27, 18 -- for {microscopy-at-msa.microscopy.com} ; Thu, 3 May 2007 07:00:41 -0500 } 27, 18 -- Received: from mobuscomm01.mail.degussa.com (uss1026.applications.degussanet.com [10.88.88.98]) } 27, 18 -- by framailout1.rz.itson.com (8.13.4/8.13.4/Debian-3sarge3) with ESMTP id l43C0ZLo027649; } 27, 18 -- Thu, 3 May 2007 14:00:36 +0200 } 27, 18 -- In-Reply-To: {200705031109.l43B9Mcd030690-at-ns.microscopy.com} } 27, 18 -- Subject: Re: [Microscopy] Optical Microscopy } 27, 18 -- To: protrain-at-emcourses.com, microscopy-at-msa.microscopy.com } 27, 18 -- X-Mailer: Lotus Notes Release 6.5.5 November 30, 2005 } 27, 18 -- Message-ID: {OFAD8E7B5A.9EF57A57-ON862572D0.0040CD9B-852572D0.0041F460-at-degussa.com} } 27, 18 -- From: frank.karl-at-degussa.com } 27, 18 -- Date: Thu, 3 May 2007 08:00:31 -0400 } 27, 18 -- X-MIMETrack: Serialize by Router on MOBUSComm01/DHexternal/US(Release 6.5.1|January 21, 2004) at } 27, 18 -- 05/03/2007 07:00:36 AM } 27, 18 -- MIME-Version: 1.0 } 27, 18 -- Content-type: text/plain; charset=US-ASCII } ==============================End of - Headers============================== }
==============================Original Headers============================== 3, 28 -- From rcmoretz-at-gmail.com Fri May 4 20:44:55 2007 3, 28 -- Received: from py-out-1112.google.com (py-out-1112.google.com [64.233.166.177]) 3, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l451is0n031145 3, 28 -- for {Microscopy-at-microscopy.com} ; Fri, 4 May 2007 20:44:55 -0500 3, 28 -- Received: by py-out-1112.google.com with SMTP id b50so889802pyh 3, 28 -- for {Microscopy-at-microscopy.com} ; Fri, 04 May 2007 18:44:54 -0700 (PDT) 3, 28 -- DKIM-Signature: a=rsa-sha1; c=relaxed/relaxed; 3, 28 -- d=gmail.com; s=beta; 3, 28 -- h=domainkey-signature:received:received:message-id:date:from:to:subject:in-reply-to:mime-version:content-type:content-transfer-encoding:content-disposition:references; 3, 28 -- b=rWEzNE/udTpwoCmEquiRPuR+YGxK4RTW9HeLUk6ZmWnIT65OSlKpGohrNC5zWXpKdqfQuFrVhSVpgdO2kDJaejuQRp0gcg66TrdZV91yDi34AEb7y5Z6sxvQDbgWks0oAjHwNfIgg7W9xzrMb07IU9Qpu4YzGqMa3owC4jJQij4= 3, 28 -- DomainKey-Signature: a=rsa-sha1; c=nofws; 3, 28 -- d=gmail.com; s=beta; 3, 28 -- h=received:message-id:date:from:to:subject:in-reply-to:mime-version:content-type:content-transfer-encoding:content-disposition:references; 3, 28 -- b=a6/RooyhuRDnppcaIkC7TjCwQRanmU5vlGm3UKogWR2RIgAL6O3u7OYcy+q896SmEz9u9crgFmLTbT6G+xnNpdhfhhUsYfdP0aIknhrPrYs9/kKkzjFgGd3TD9s2dyuVSvFW7n/Y9l5WVy+plHBLmPmnYtE53wulngsYnd4Wwa4= 3, 28 -- Received: by 10.65.152.17 with SMTP id e17mr6940506qbo.1178329494131; 3, 28 -- Fri, 04 May 2007 18:44:54 -0700 (PDT) 3, 28 -- Received: by 10.64.196.7 with HTTP; Fri, 4 May 2007 18:44:54 -0700 (PDT) 3, 28 -- Message-ID: {950e3cfd0705041844j116c62bhe993d2e932a7e886-at-mail.gmail.com} 3, 28 -- Date: Fri, 4 May 2007 21:44:54 -0400 3, 28 -- From: "Roger Moretz" {rcmoretz-at-gmail.com} 3, 28 -- To: frank.karl-at-degussa.com, "Microscopy Listserv" {Microscopy-at-microscopy.com} 3, 28 -- Subject: Re: [Microscopy] Re: Optical Microscopy 3, 28 -- In-Reply-To: {200705031205.l43C51u5027698-at-ns.microscopy.com} 3, 28 -- MIME-Version: 1.0 3, 28 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 3, 28 -- Content-Transfer-Encoding: 7bit 3, 28 -- Content-Disposition: inline 3, 28 -- References: {200705031205.l43C51u5027698-at-ns.microscopy.com} ==============================End of - Headers==============================
I think Shu has a very good point, the explanation I had been given 40 years ago that I sent through to him was:-
"Yes it is a very strange phenomena. Remember the plastic film, like any material in the electron beam, has a charge upon it, even so close to the copper grid. People like David Joy will tell you that even the copper grid has a charge upon it, nothing is a perfect conductor! The changes in texture/thickness within the film take equal charge, but like charges repel so when the charge reaches a specific level repulsion of two areas eventually tears the film apart along the weaker areas. That is the explanation that I was told many years ago, it does seem to fit the action that occurs. Reduce the number of incident electrons and you reduce the charge, coat with carbon and you increase the thickness and the conductivity but you could spoil the resolution in relation to T/10.
For some reason the backscatter from the aperture reduces this charge effect, or spraying electrons onto the sample with a charge neutraliser (available in the 60s) did the same! The grid provides conductivity, the backscatter seems to provide charge neutralisation, it is the charge that breaks the film."
Just shows once again how the listserver can be so informative - "don't ask and you don't get!"
Steve Chapman Senior Consultant Protrain For electron microscopy consultancy and training world wide Tel +44 1280 816512 Fax +44 1280 814007 Mobile +44 7711 606967 Web www.emcourses.com
----- Original Message ----- X-from: {shu-at-caltech.edu} To: {protrain-at-emcourses.com} Sent: Friday, May 04, 2007 9:19 PM
Hi, Jan- The question of the redox potential also puzzled me when I first came up with this explanation for why membranes get "bleached" after fixation with OsO4. Your idea of the alkalinity making a difference is an appealing alternative explanation. Another aspect of this phenomenon (problem, I mean) that was hard to understand was that it appeared to be restricted to tissue culture preparations, where the sample is very thin. Would the alkalinity affect the thin sample preferentially? Using the same reagents, we had no problem with the apparent extraction of OsO4 from membranes in bulk tissues.
Whatever we did, the hypothesis I gave was merely based on the empirical finding that changing the solvent solved the problem. It has no implications for the true mechanism of this effect. Now that there are so many new instrumental methods, people are making progress on definfing the chemical after-effects of fixation methods! With best regards, Carol
==============================Original Headers============================== 4, 14 -- From heckman-at-bgnet.bgsu.edu Sat May 5 15:41:30 2007 4, 14 -- Received: from smtp01.bgsu.edu (smtp01.bgsu.edu [129.1.5.17]) 4, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l45KfUPi003088 4, 14 -- for {microscopy-at-microscopy.com} ; Sat, 5 May 2007 15:41:30 -0500 4, 14 -- Received: from localhost (webmail.bgsu.edu [129.1.5.21]) 4, 14 -- by smtp01.bgsu.edu (Switch-3.2.5/Switch-3.1.6) with ESMTP id l45KfOEw025052; 4, 14 -- Sat, 5 May 2007 16:41:24 -0400 (EDT) 4, 14 -- MIME-Version: 1.0 4, 14 -- Date: Sat, 5 May 2007 16:41:25 -0400 4, 14 -- From: "heckman-at-bgnet.bgsu.edu" {heckman-at-bgnet.bgsu.edu} 4, 14 -- Message-Id: {1178397685-15923.00016.00613-smmsdV2.1.6-at-smtp.bgsu.edu} 4, 14 -- X-SMMS-Source: 72.240.89.131 4, 14 -- To: microscopy-at-microscopy.com, {leunissen-at-aurion.nl} 4, 14 -- Subject: Re: [Microscopy] Help with enhancing contrast of Mitochondrial ==============================End of - Headers==============================
Whilst we have been discussing the failure of support films in TEM and possible ionisation taking place, I feel it is right to indicate another form of problem?
If there is a leak near to the specimen area of the TEM, usually the specimen exchange mechanism, another interesting problem may appear. Whilst working with a thin support film you may notice that the film slowly becomes lacy, the holes growing with time until the film falls apart. What is happening is the gas from the leak is being ionised and it is etching away the support film. From my service background it was a give-away as a specimen exchange leak! This usually showed up whilst I was checking the instrument with a holey carbon film thus demonstrating that a holey film may come in handy for more than image checking (?).
Hope this helps one of you in the future?
Regards
Steve Chapman Senior Consultant Protrain For electron microscopy consultancy and training world wide Tel +44 1280 816512 Fax +44 1280 814007 Mobile +44 7711 606967 Web www.emcourses.com
==============================Original Headers============================== 7, 26 -- From protrain-at-emcourses.com Sun May 6 04:39:06 2007 7, 26 -- Received: from smtp2.pri.skybb.uk.easynet.net (smtp2.pri.skybb.uk.easynet.net [87.86.189.70]) 7, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l469d5Zr011160 7, 26 -- for {microscopy-at-microscopy.com} ; Sun, 6 May 2007 04:39:06 -0500 7, 26 -- Received: from [90.203.67.208] (helo=advent) 7, 26 -- by smtp2.pri.skybb.uk.easynet.net with smtp (Exim 4.62) 7, 26 -- (envelope-from {protrain-at-emcourses.com} ) 7, 26 -- id 1HkdCq-0001FO-HM 7, 26 -- for microscopy-at-microscopy.com; Sun, 06 May 2007 10:39:04 +0100 7, 26 -- Message-ID: {000601c78fc3$02178900$0200a8c0-at-advent} 7, 26 -- Reply-To: "Steve Chapman" {protrain-at-emcourses.com} 7, 26 -- From: "Steve Chapman" {protrain-at-emcourses.com} 7, 26 -- To: "American Soc" {microscopy-at-microscopy.com} 7, 26 -- Subject: [Microscopy] Support Film Damage in TEM 7, 26 -- Date: Sun, 6 May 2007 10:43:19 +0100 7, 26 -- Organization: Protrain 7, 26 -- MIME-Version: 1.0 7, 26 -- Content-Type: text/plain; 7, 26 -- format=flowed; 7, 26 -- charset="iso-8859-1"; 7, 26 -- reply-type=original 7, 26 -- Content-Transfer-Encoding: 7bit 7, 26 -- X-Priority: 3 7, 26 -- X-MSMail-Priority: Normal 7, 26 -- X-Mailer: Microsoft Outlook Express 6.00.2900.3028 7, 26 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 ==============================End of - Headers==============================
I am looking for a carbon rod sharpened for rods with diameter of 0.12 inch diameter rods. We had a great electrical one that rotated the rod and you just moved a small cutting edge along a track to cut a nice long and narrow rod at the end of the larger rod. The rotating shaft no longer is stable so rods break too easily.
Would appreciate advice as to replacements and would prefer electrical one rather than the hand-held type. =================================================================== SPI Supplies has offered an electrically driven carbon rod sharpener for a number of years, see URL http://www.2spi.com/catalog/spec_prep/carbon-rod-sharpener.shtml
for a photo of the product and full technical description.
Our standard set up for 1/8" diameter or 3 mm should work for your 0.12" rods. The product is shipped with what would be needed if you have carbon rods of some other diameter such as 4.8 and 6.1 mm diameter. In that sense it is a "universal" sharpener and can sharpen all diameters of rods found in an EM laboratory.
Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
==============================Original Headers============================== 11, 25 -- From cgarber-at-2spi.com Sun May 6 10:14:35 2007 11, 25 -- Received: from diskless5.axs2000.net (mail.axs2000.net [209.120.196.43]) 11, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l46FEWG1019491 11, 25 -- for {microscopy-at-microscopy.com} ; Sun, 6 May 2007 10:14:35 -0500 11, 25 -- Received: from webmail.idv.net (diskless6.external [209.120.196.50]) 11, 25 -- (authenticated bits=0) 11, 25 -- by diskless5.axs2000.net (8.12.11/8.12.11) with ESMTP id l46FEWnQ021981 11, 25 -- for {microscopy-at-microscopy.com} ; Sun, 6 May 2007 11:14:32 -0400 11, 25 -- X-IDV-FirstRcvd: diskless6.external [209.120.196.50] 11, 25 -- X-IDV-HELO: webmail.idv.net 11, 25 -- X-IDV-Authenticated-User: cgarber 11, 25 -- Received: from 68.246.64.184 (auth. user cgarber-at-mail.2spi.com) 11, 25 -- by webmail.idv.net with HTTP; Sun, 06 May 2007 10:14:32 -0500 11, 25 -- To: microscopy-at-microscopy.com 11, 25 -- Subject: Carbon rod sharpener 11, 25 -- Date: Sun, 06 May 2007 10:14:32 -0500 11, 25 -- X-Mailer: IlohaMail/0.9.0 (On: webmail.idv.net) 11, 25 -- Message-ID: {IXc2edOt.1178464472.7084200.cgarber-at-mail.2spi.com} 11, 25 -- From: {cgarber-at-2spi.com} 11, 25 -- Bounce-To: {cgarber-at-2spi.com} 11, 25 -- Errors-To: {cgarber-at-2spi.com} 11, 25 -- MIME-Version: 1.0 11, 25 -- Content-Type: text/plain; charset=ISO-8859-1 11, 25 -- Content-Transfer-Encoding: 8bit 11, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l46FEWG1019491 ==============================End of - Headers==============================
I would like to thank all the people who answered and help me to understand what was going on. After two days of checking it seems that our filament was not properly centered in the Wehnelt. Best Regards Patrizia
-- ______________________________________________ Patrizia Canton PhD Dept. of Physical Chemistry Via Torino 155/b I-30170 Venezia-Mestre Italy Phone +39-041-2346790 Fax +39-041-2346747 e-mail cantonpa-at-unive.it ______________________________________________
==============================Original Headers============================== 3, 26 -- From cantonpa-at-unive.it Mon May 7 08:27:57 2007 3, 26 -- Received: from zeus.unive.it (zeus.unive.it [157.138.1.81]) 3, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l47DRuRv023174 3, 26 -- for {Microscopy-at-microscopy.com} ; Mon, 7 May 2007 08:27:57 -0500 3, 26 -- Received: from localhost (localhost.localdomain [127.0.0.1]) 3, 26 -- by zeus.unive.it (8.12.11.20060308/8.12.11) with ESMTP id l47DHwVh024586 3, 26 -- for {Microscopy-at-microscopy.com} ; Mon, 7 May 2007 15:17:58 +0200 3, 26 -- X-Virus-Scanned: amavisd-new at unive.it 3, 26 -- Received: from zeus.unive.it ([127.0.0.1]) 3, 26 -- by localhost (zeus.unive.it [127.0.0.1]) (amavisd-new, port 10024) 3, 26 -- with LMTP id rJMraECwSQ-J for {Microscopy-at-microscopy.com} ; 3, 26 -- Mon, 7 May 2007 15:17:57 +0200 (CEST) 3, 26 -- Received: from fibre7.unive.it (fibre7.unive.it [157.138.8.7]) 3, 26 -- by zeus.unive.it (8.12.11.20060308/8.12.11) with ESMTP id l47DHsah024531; 3, 26 -- Mon, 7 May 2007 15:17:54 +0200 3, 26 -- Received: from localhost (cantonpa-at-localhost) 3, 26 -- by fibre7.unive.it (8.12.11.20060308/8.12.11) with ESMTP id l47DQf9N024385; 3, 26 -- Mon, 7 May 2007 15:26:41 +0200 3, 26 -- X-Authentication-Warning: fibre7.unive.it: cantonpa owned process doing -bs 3, 26 -- Date: Mon, 7 May 2007 15:26:41 +0200 (CEST) 3, 26 -- From: Patrizia Canton {cantonpa-at-unive.it} 3, 26 -- To: Microscopy-at-microscopy.com 3, 26 -- Subject: LaB6 filament:thanks 3, 26 -- Message-ID: {Pine.LNX.4.44.0705071523430.23203-100000-at-fibre7.unive.it} 3, 26 -- MIME-Version: 1.0 3, 26 -- Content-Type: TEXT/PLAIN; charset=US-ASCII ==============================End of - Headers==============================
Dear Justin, This sounds like a very difficult problem to solve, especially if you don't have a good high vacuum gauge. I would suggest you check the diffusion pump and make sure it is getting hot at the bottom and cool at the top and is staying that way. I once had an "intermittent vacuum leak" that turned out to be that I had put the wrong diffusion pump oil in the pump and it was not quite boiling. That only caused poor high vacuum, though, not degradation back to atmosphere. Also check the quantity and quality (color) of the oil in the diffusion pump. Also, some sensor or vacuum gauge may be dirty or faulty and switching the vacuum off intermittently. I see no choice but to sit there until it does the bad thing while you are watching. Good luck,
-----Original Message----- X-from: kraftpiano-at-gmail.com [mailto:kraftpiano-at-gmail.com] Sent: May 4, 2007 6:34 PM To: mager-at-interchange.ubc.ca
I'm completely dumbfounded by this one. I am in the process of checking for leaks on our SEM, and it pumps down perfectly one moment, then it won't even trigger PiG3. When I start to take things apart to check seals and such, then I put it back together (After finding nothing wrong) it evacuates perfectly. Then I let it sit overnight to see if there is a leak, and when I get in the next morning, it is completely at atmospheric pressure and then won't hold a vacuum and trigger PiG3 anymore. I'm a little confused at how it will hold a vacuum only the first time I pump it down after dismantling and reassembling parts of the vacuum system. I don't have a leak detector, but boy would I love one! (I also don't have any gauges capable of measuring absolute vacuum vs. relative vacuum, so I have no idea what the actual measurement in torr is.)
--Justin A. Kraft
==============================Original Headers============================== 2, 26 -- From kraftpiano-at-gmail.com Fri May 4 20:28:40 2007 2, 26 -- Received: from wr-out-0506.google.com (wr-out-0506.google.com [64.233.184.234]) 2, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l451Seww019490 2, 26 -- for {microscopy-at-microscopy.com} ; Fri, 4 May 2007 20:28:40 -0500 2, 26 -- Received: by wr-out-0506.google.com with SMTP id i31so1164508wra 2, 26 -- for {microscopy-at-microscopy.com} ; Fri, 04 May 2007 18:28:39 -0700 (PDT) 2, 26 -- DKIM-Signature: a=rsa-sha1; c=relaxed/relaxed; 2, 26 -- d=gmail.com; s=beta; 2, 26 -- h=domainkey-signature:received:received:message-id:date:from:to:subject:mime -version:content-type:content-transfer-encoding:content-disposition; 2, 26 -- b=cJmt3MZGjC95glyyFu+AdPFMqPSYoAxgMR+EEmmwxcGSmf9jAaZNhlAbKhDp8Lw0pkV58J8V4X Unfo2K8+l5Y4lPkodDfOgmAT8bDZwXxnh/Qw5RlVm+u2fQE07ZLgT4B1VLT6ZP9fhkPJG4MkB4qF E4yS/976P3LuVwAtAhZQ0= 2, 26 -- DomainKey-Signature: a=rsa-sha1; c=nofws; 2, 26 -- d=gmail.com; s=beta; 2, 26 -- h=received:message-id:date:from:to:subject:mime-version:content-type:content -transfer-encoding:content-disposition; 2, 26 -- b=j+NcTQR1YAWPfjXoMiGt0pCoHrffvabybEPYQGQALUh1uQocYfxunYiU3EnEnAAKmVjNCewAQ6 SdNwXlbQMuD1iulfnngZH6DMhCKSh39cDDtCMhY/6cv+EVn4fQYR/uUglWyJ9NUB9r0JIk2UtZwV sOBnAxfzwvzLYLfnzir0s= 2, 26 -- Received: by 10.114.133.1 with SMTP id g1mr1349202wad.1178328519224; 2, 26 -- Fri, 04 May 2007 18:28:39 -0700 (PDT) 2, 26 -- Received: by 10.114.79.12 with HTTP; Fri, 4 May 2007 18:28:38 -0700 (PDT) 2, 26 -- Message-ID: {25e2b0d20705041828v19b4f286r2878534d132d125-at-mail.gmail.com} 2, 26 -- Date: Fri, 4 May 2007 21:28:38 -0400 2, 26 -- From: "Justin Kraft" {kraftpiano-at-gmail.com} 2, 26 -- To: microscopy-at-microscopy.com 2, 26 -- Subject: SEM: Intermittent Vacuum Leak 2, 26 -- MIME-Version: 1.0 2, 26 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 2, 26 -- Content-Transfer-Encoding: 7bit 2, 26 -- Content-Disposition: inline ==============================End of - Headers==============================
==============================Original Headers============================== 10, 35 -- From mager-at-interchange.ubc.ca Mon May 7 11:12:37 2007 10, 35 -- Received: from mr8.mail-relay.ubc.ca (mr8.mail-relay.ubc.ca [137.82.45.15]) 10, 35 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l47GCaIi005664 10, 35 -- for {microscopy-at-microscopy.com} ; Mon, 7 May 2007 11:12:37 -0500 10, 35 -- Received: from mr8.mail-relay.ubc.ca (localhost [127.0.0.1]) 10, 35 -- by localhost (Postfix) with SMTP id 59B2810AC2 10, 35 -- for {microscopy-at-microscopy.com} ; Mon, 7 May 2007 09:12:36 -0700 (PDT) 10, 35 -- Received: from mta1.interchange.ubc.ca (mta1.interchange.ubc.ca [142.103.145.69]) 10, 35 -- by mr8.mail-relay.ubc.ca (Postfix) with ESMTP 10, 35 -- for {microscopy-at-microscopy.com} ; Mon, 7 May 2007 09:12:35 -0700 (PDT) 10, 35 -- Received: from Mary (desk.staff.mmat.ubc.ca [137.82.16.178]) 10, 35 -- by smtp.interchange.ubc.ca 10, 35 -- (iPlanet Messaging Server 5.2 HotFix 1.21 (built Sep 8 2003)) 10, 35 -- with ESMTPS id {0JHO001C7ICZ7V-at-smtp.interchange.ubc.ca} for 10, 35 -- microscopy-at-microscopy.com; Mon, 07 May 2007 09:12:35 -0700 (PDT) 10, 35 -- Date: Mon, 07 May 2007 09:12:10 -0700 10, 35 -- From: Mary Mager {mager-at-interchange.ubc.ca} 10, 35 -- Subject: RE: [Microscopy] SEM: Intermittent Vacuum Leak 10, 35 -- In-reply-to: {200705050134.l451YBlU028241-at-ns.microscopy.com} 10, 35 -- To: kraftpiano-at-gmail.com 10, 35 -- Cc: "'Microscopy'" {microscopy-at-microscopy.com} 10, 35 -- Reply-to: mager-at-interchange.ubc.ca 10, 35 -- Message-id: {0JHO001C8ICZ7V-at-smtp.interchange.ubc.ca} 10, 35 -- Organization: Materials Eng. UBC 10, 35 -- MIME-version: 1.0 10, 35 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 10, 35 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 10, 35 -- Content-type: text/plain; charset=US-ASCII 10, 35 -- Content-transfer-encoding: 7bit 10, 35 -- Thread-index: AceOtX4m59s7fJe9Rvm4l1lQyAtHEQCC8eIg 10, 35 -- X-UBC-Scanned: Sophos PureMessage 5.3.1.294258, Antispam-Engine: 2.5.0.283055, Antispam-Data: 2007.5.7.85134 10, 35 -- X-UBC-Relayed: Relayed through mail-relay.ubc.ca 10, 35 -- X-PerlMx-Spam: Probability=7%, Report=__C230066_P5 0, __CP_NAME_BODY 0, __CP_URI_IN_BODY 0, __CT 0, __CTE 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __HAS_X_MAILER 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0, __STOCK_PHRASE_24 0, __USER_AGENT_MS_GENERIC 0 10, 35 -- X-Spam-Level: 10, 35 -- X-Spam-Flag: No ==============================End of - Headers==============================
Does it truly not have vacuum, or only indicate this?
A possibility is that the vacuum sensors or processing circuits are giving erratic performance/faulty response and since the control of the vacuum system is based on these it can complicate the troubleshooting.
Also, most scopes have a "Klixon" on the ODP column (or 2: one for overtemp and one for "hot enough to function") that are continually subjected to heat and will go flakey in time and may be shutting down the ODP. We've had a good bit of this problem over the years.
Try to get a handle on the vacuum by some independent means if possible - if not a sepatate gage unit, then try to read the existing raw sensor voltage where it connects to the microscope to see what is happening (this assumes you have/read schematics and can locate the places to read the sensor); or your scope may have some status LEDs on the vacuum control board that might actually be labelled?
If don't want to pay for a commercial Pirani unit and you are inclined to "tinkering" I have some information on a DIY "Light Bulb Pirani" gage sensor/circuit based on an opened light bulb tungsten element and a simple driver circuit. The tungsten is heated to a preset temperature and the voltage required to maintain that temperature is proportional to pressure - at atmosphere the heat is conducted away so more voltage is required; there is a spreadsheet with data, a schematic, and a couple of photos. This system is not temperature compensated, so is best kept away from sources of heat and for relative vacuum level sensing you probably don't need to calibrate against another gage, just look at the spreadsheet data. It requires only 2 feedthrough pins into the vacuum.
mager-at-interchange.ubc.ca wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Dear Justin, } This sounds like a very difficult problem to solve, especially if you don't } have a good high vacuum gauge. I would suggest you check the diffusion pump } and make sure it is getting hot at the bottom and cool at the top and is } staying that way. I once had an "intermittent vacuum leak" that turned out } to be that I had put the wrong diffusion pump oil in the pump and it was not } quite boiling. That only caused poor high vacuum, though, not degradation } back to atmosphere. Also check the quantity and quality (color) of the oil } in the diffusion pump. Also, some sensor or vacuum gauge may be dirty or } faulty and switching the vacuum off intermittently. I see no choice but to } sit there until it does the bad thing while you are watching. } Good luck, } } Mary Fletcher (nee Mager) } Electron Microscopist } Materials Eng. UBC } #309 - 6350 Stores Road } Vancouver, B.C. V6T 1Z4 } Canada } Tel: 604-822-5648 } Fax: 604-822-3619 } email: mager-at-interchange.ubc.ca } } -----Original Message----- } X-from: kraftpiano-at-gmail.com [mailto:kraftpiano-at-gmail.com] } Sent: May 4, 2007 6:34 PM } To: mager-at-interchange.ubc.ca } Subject: [Microscopy] SEM: Intermittent Vacuum Leak } } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } I'm completely dumbfounded by this one. I am in the process of } checking for leaks on our SEM, and it pumps down perfectly one moment, } then it won't even trigger PiG3. When I start to take things apart to } check seals and such, then I put it back together (After finding } nothing wrong) it evacuates perfectly. Then I let it sit overnight to } see if there is a leak, and when I get in the next morning, it is } completely at atmospheric pressure and then won't hold a vacuum and } trigger PiG3 anymore. I'm a little confused at how it will hold a } vacuum only the first time I pump it down after dismantling and } reassembling parts of the vacuum system. I don't have a leak } detector, but boy would I love one! (I also don't have any gauges } capable of measuring absolute vacuum vs. relative vacuum, so I have no } idea what the actual measurement in torr is.) } } --Justin A. Kraft } } ==============================Original Headers============================== } 2, 26 -- From kraftpiano-at-gmail.com Fri May 4 20:28:40 2007 } 2, 26 -- Received: from wr-out-0506.google.com (wr-out-0506.google.com } [64.233.184.234]) } 2, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } l451Seww019490 } 2, 26 -- for {microscopy-at-microscopy.com} ; Fri, 4 May 2007 20:28:40 } -0500 } 2, 26 -- Received: by wr-out-0506.google.com with SMTP id i31so1164508wra } 2, 26 -- for {microscopy-at-microscopy.com} ; Fri, 04 May 2007 18:28:39 } -0700 (PDT) } 2, 26 -- DKIM-Signature: a=rsa-sha1; c=relaxed/relaxed; } 2, 26 -- d=gmail.com; s=beta; } 2, 26 -- } h=domainkey-signature:received:received:message-id:date:from:to:subject:mime } -version:content-type:content-transfer-encoding:content-disposition; } 2, 26 -- } b=cJmt3MZGjC95glyyFu+AdPFMqPSYoAxgMR+EEmmwxcGSmf9jAaZNhlAbKhDp8Lw0pkV58J8V4X } Unfo2K8+l5Y4lPkodDfOgmAT8bDZwXxnh/Qw5RlVm+u2fQE07ZLgT4B1VLT6ZP9fhkPJG4MkB4qF } E4yS/976P3LuVwAtAhZQ0= } 2, 26 -- DomainKey-Signature: a=rsa-sha1; c=nofws; } 2, 26 -- d=gmail.com; s=beta; } 2, 26 -- } h=received:message-id:date:from:to:subject:mime-version:content-type:content } -transfer-encoding:content-disposition; } 2, 26 -- } b=j+NcTQR1YAWPfjXoMiGt0pCoHrffvabybEPYQGQALUh1uQocYfxunYiU3EnEnAAKmVjNCewAQ6 } SdNwXlbQMuD1iulfnngZH6DMhCKSh39cDDtCMhY/6cv+EVn4fQYR/uUglWyJ9NUB9r0JIk2UtZwV } sOBnAxfzwvzLYLfnzir0s= } 2, 26 -- Received: by 10.114.133.1 with SMTP id } g1mr1349202wad.1178328519224; } 2, 26 -- Fri, 04 May 2007 18:28:39 -0700 (PDT) } 2, 26 -- Received: by 10.114.79.12 with HTTP; Fri, 4 May 2007 18:28:38 -0700 } (PDT) } 2, 26 -- Message-ID: } {25e2b0d20705041828v19b4f286r2878534d132d125-at-mail.gmail.com} } 2, 26 -- Date: Fri, 4 May 2007 21:28:38 -0400 } 2, 26 -- From: "Justin Kraft" {kraftpiano-at-gmail.com} } 2, 26 -- To: microscopy-at-microscopy.com } 2, 26 -- Subject: SEM: Intermittent Vacuum Leak } 2, 26 -- MIME-Version: 1.0 } 2, 26 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed } 2, 26 -- Content-Transfer-Encoding: 7bit } 2, 26 -- Content-Disposition: inline } ==============================End of - Headers============================== } } } ==============================Original Headers============================== } 10, 35 -- From mager-at-interchange.ubc.ca Mon May 7 11:12:37 2007 } 10, 35 -- Received: from mr8.mail-relay.ubc.ca (mr8.mail-relay.ubc.ca [137.82.45.15]) } 10, 35 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l47GCaIi005664 } 10, 35 -- for {microscopy-at-microscopy.com} ; Mon, 7 May 2007 11:12:37 -0500 } 10, 35 -- Received: from mr8.mail-relay.ubc.ca (localhost [127.0.0.1]) } 10, 35 -- by localhost (Postfix) with SMTP id 59B2810AC2 } 10, 35 -- for {microscopy-at-microscopy.com} ; Mon, 7 May 2007 09:12:36 -0700 (PDT) } 10, 35 -- Received: from mta1.interchange.ubc.ca (mta1.interchange.ubc.ca [142.103.145.69]) } 10, 35 -- by mr8.mail-relay.ubc.ca (Postfix) with ESMTP } 10, 35 -- for {microscopy-at-microscopy.com} ; Mon, 7 May 2007 09:12:35 -0700 (PDT) } 10, 35 -- Received: from Mary (desk.staff.mmat.ubc.ca [137.82.16.178]) } 10, 35 -- by smtp.interchange.ubc.ca } 10, 35 -- (iPlanet Messaging Server 5.2 HotFix 1.21 (built Sep 8 2003)) } 10, 35 -- with ESMTPS id {0JHO001C7ICZ7V-at-smtp.interchange.ubc.ca} for } 10, 35 -- microscopy-at-microscopy.com; Mon, 07 May 2007 09:12:35 -0700 (PDT) } 10, 35 -- Date: Mon, 07 May 2007 09:12:10 -0700 } 10, 35 -- From: Mary Mager {mager-at-interchange.ubc.ca} } 10, 35 -- Subject: RE: [Microscopy] SEM: Intermittent Vacuum Leak } 10, 35 -- In-reply-to: {200705050134.l451YBlU028241-at-ns.microscopy.com} } 10, 35 -- To: kraftpiano-at-gmail.com } 10, 35 -- Cc: "'Microscopy'" {microscopy-at-microscopy.com} } 10, 35 -- Reply-to: mager-at-interchange.ubc.ca } 10, 35 -- Message-id: {0JHO001C8ICZ7V-at-smtp.interchange.ubc.ca} } 10, 35 -- Organization: Materials Eng. UBC } 10, 35 -- MIME-version: 1.0 } 10, 35 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 } 10, 35 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 } 10, 35 -- Content-type: text/plain; charset=US-ASCII } 10, 35 -- Content-transfer-encoding: 7bit } 10, 35 -- Thread-index: AceOtX4m59s7fJe9Rvm4l1lQyAtHEQCC8eIg } 10, 35 -- X-UBC-Scanned: Sophos PureMessage 5.3.1.294258, Antispam-Engine: 2.5.0.283055, Antispam-Data: 2007.5.7.85134 } 10, 35 -- X-UBC-Relayed: Relayed through mail-relay.ubc.ca } 10, 35 -- X-PerlMx-Spam: Probability=7%, Report=__C230066_P5 0, __CP_NAME_BODY 0, __CP_URI_IN_BODY 0, __CT 0, __CTE 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __HAS_X_MAILER 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0, __STOCK_PHRASE_24 0, __USER_AGENT_MS_GENERIC 0 } 10, 35 -- X-Spam-Level: } 10, 35 -- X-Spam-Flag: No } ==============================End of - Headers==============================
==============================Original Headers============================== 10, 21 -- From dac-at-research.umass.edu Mon May 7 12:13:36 2007 10, 21 -- Received: from race5.oit.umass.edu (race5.oit.umass.edu [128.119.101.41]) 10, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l47HDYVa018299 10, 21 -- for {Microscopy-at-microscopy.com} ; Mon, 7 May 2007 12:13:35 -0500 10, 21 -- Received: from [172.30.55.164] (eutopia.bio.umass.edu [128.119.55.30]) 10, 21 -- (authenticated bits=0) 10, 21 -- by race5.oit.umass.edu (8.13.7/8.13.7) with ESMTP id l47HDXNO020491 10, 21 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NOT) 10, 21 -- for {Microscopy-at-microscopy.com} ; Mon, 7 May 2007 13:13:33 -0400 10, 21 -- Message-ID: {463F6CA6.1050208-at-research.umass.edu} 10, 21 -- Date: Mon, 07 May 2007 13:15:02 -0500 10, 21 -- From: Dale Callaham {dac-at-research.umass.edu} 10, 21 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.0; en-US; rv:1.8.1.2) Gecko/20070222 SeaMonkey/1.1.1 10, 21 -- MIME-Version: 1.0 10, 21 -- To: Microscopy Listserver {Microscopy-at-microscopy.com} 10, 21 -- Subject: Re: [Microscopy] RE: SEM: Intermittent Vacuum Leak 10, 21 -- References: {200705071618.l47GIeDd015421-at-ns.microscopy.com} 10, 21 -- In-Reply-To: {200705071618.l47GIeDd015421-at-ns.microscopy.com} 10, 21 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 10, 21 -- Content-Transfer-Encoding: 7bit 10, 21 -- X-Whitelist: TRUE ==============================End of - Headers==============================
I do not know which instrument that you have as you do not say? I have experienced a problem in the past where the customer behaved exactly as you have. Strip and rebuild no problem, next day no good! In this case simply turning the vacuum system off and on overcame the fault. Could this be your problem as when you strip and rebuild you turn the system off?
If you could provide details of which instrument that you are using and how it is pumped - diffusion pump or turbo molecular - this would help?
Please get back and I will try to help.
Steve Chapman Senior Consultant Protrain For electron microscopy consultancy and training world wide Tel +44 1280 816512 Fax +44 1280 814007 Mobile +44 7711 606967 Web www.emcourses.com
----- Original Message ----- X-from: {kraftpiano-at-gmail.com} To: {protrain-at-emcourses.com} Sent: Saturday, May 05, 2007 2:29 AM
Vacuum leak update:
Thanks to the many suggestions I received, the vacuum leak has been identified. I thought I'd send in a summary of the suggestions that ended up being the most helpful so those with similar problems in the future may benefit. Here is the summary:
1: Using a large quantity of various sized stoppers, isolate different parts of the vacuum system to determine where the leak may be.
2: If the system has multiple piranni gauges (As the JSM-840 does) try switching the gauges around to make sure that it is not a bad gauge.
3: Check that the diffusion pumps are heating up. (Although this was not the issue with mine, since it was not reaching a vacuum level that would allow the diffusion pumps to begin to work, I thought I'd include it just as general reference.) Also check to make sure that the temperature sensors on the diffusion pumps are functional.
4: Short of a leak detector, spray some acetone or high purity isopropyl alcohol on the different seals. Wait a few minutes after each spray. If there is a leak in that spot, then the vacuum will increase momentarily, then go back down as the alcohol or acetone plugs the leak and then vaporizes on the inside of the system. Keep doing this moving from the outermost portions of the vacuum system to the pump connection until you find it.
5: Pump the system down as far as it will go, then seal all of the valves in the closed position. Wait. The next morning, the section with the leak will not have a vacuum in it, but the others will.
Rick Becker given this handy suggestion for repairing the leak when found:
If the leak is in a section that you can get around (i.e. the junction between two column sections) standard electrical tape can repair it temporarily. Make sure it is high quality electrical tape. The thicker, the better. Decent electrical tape can hold a vacuum of 10 e-9 torr. Not bad- the tape is now holding the vacuum between the "T" fitting under the chamber (Bottom is the small vent valve, top is the chamber, and the leg on the side is a large butterfly valve into the rest of the vacuum system) and the large butterfly valve. I ordered a new gasket from Small Parts, Inc. (They really do have everything!) but in the mean time, it's holding fine.
As it turns out (For those of you wondering) when we re-assembled the vacuum system after carrying it piece by piece up the stairs, we must have accidentally pinched the gasket when putting the two fittings together. This caused a nick about the size of a couple of grains of sand in the gasket, which would become a problem after being reassembled when the gasket would roll slightly after tightening the screws.
Hope this summary helps others who might need it!
--Justin.
On 5/4/07, kraftpiano-at-gmail.com {kraftpiano-at-gmail.com} wrote: } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } I'm completely dumbfounded by this one. I am in the process of } checking for leaks on our SEM, and it pumps down perfectly one moment, } then it won't even trigger PiG3. When I start to take things apart to } check seals and such, then I put it back together (After finding } nothing wrong) it evacuates perfectly. Then I let it sit overnight to } see if there is a leak, and when I get in the next morning, it is } completely at atmospheric pressure and then won't hold a vacuum and } trigger PiG3 anymore. I'm a little confused at how it will hold a } vacuum only the first time I pump it down after dismantling and } reassembling parts of the vacuum system. I don't have a leak } detector, but boy would I love one! (I also don't have any gauges } capable of measuring absolute vacuum vs. relative vacuum, so I have no } idea what the actual measurement in torr is.) } } --Justin A. Kraft } } ==============================Original Headers============================== } 2, 26 -- From kraftpiano-at-gmail.com Fri May 4 20:28:40 2007 } 2, 26 -- Received: from wr-out-0506.google.com (wr-out-0506.google.com [64.233.184.234]) } 2, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l451Seww019490 } 2, 26 -- for {microscopy-at-microscopy.com} ; Fri, 4 May 2007 20:28:40 -0500 } 2, 26 -- Received: by wr-out-0506.google.com with SMTP id i31so1164508wra } 2, 26 -- for {microscopy-at-microscopy.com} ; Fri, 04 May 2007 18:28:39 -0700 (PDT) } 2, 26 -- DKIM-Signature: a=rsa-sha1; c=relaxed/relaxed; } 2, 26 -- d=gmail.com; s=beta; } 2, 26 -- h=domainkey-signature:received:received:message-id:date:from:to:subject:mime-version:content-type:content-transfer-encoding:content-disposition; } 2, 26 -- b=cJmt3MZGjC95glyyFu+AdPFMqPSYoAxgMR+EEmmwxcGSmf9jAaZNhlAbKhDp8Lw0pkV58J8V4XUnfo2K8+l5Y4lPkodDfOgmAT8bDZwXxnh/Qw5RlVm+u2fQE07ZLgT4B1VLT6ZP9fhkPJG4MkB4qFE4yS/976P3LuVwAtAhZQ0= } 2, 26 -- DomainKey-Signature: a=rsa-sha1; c=nofws; } 2, 26 -- d=gmail.com; s=beta; } 2, 26 -- h=received:message-id:date:from:to:subject:mime-version:content-type:content-transfer-encoding:content-disposition; } 2, 26 -- b=j+NcTQR1YAWPfjXoMiGt0pCoHrffvabybEPYQGQALUh1uQocYfxunYiU3EnEnAAKmVjNCewAQ6SdNwXlbQMuD1iulfnngZH6DMhCKSh39cDDtCMhY/6cv+EVn4fQYR/uUglWyJ9NUB9r0JIk2UtZwVsOBnAxfzwvzLYLfnzir0s= } 2, 26 -- Received: by 10.114.133.1 with SMTP id g1mr1349202wad.1178328519224; } 2, 26 -- Fri, 04 May 2007 18:28:39 -0700 (PDT) } 2, 26 -- Received: by 10.114.79.12 with HTTP; Fri, 4 May 2007 18:28:38 -0700 (PDT) } 2, 26 -- Message-ID: {25e2b0d20705041828v19b4f286r2878534d132d125-at-mail.gmail.com} } 2, 26 -- Date: Fri, 4 May 2007 21:28:38 -0400 } 2, 26 -- From: "Justin Kraft" {kraftpiano-at-gmail.com} } 2, 26 -- To: microscopy-at-microscopy.com } 2, 26 -- Subject: SEM: Intermittent Vacuum Leak } 2, 26 -- MIME-Version: 1.0 } 2, 26 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed } 2, 26 -- Content-Transfer-Encoding: 7bit } 2, 26 -- Content-Disposition: inline } ==============================End of - Headers============================== }
==============================Original Headers============================== 13, 28 -- From kraftpiano-at-gmail.com Mon May 7 15:31:42 2007 13, 28 -- Received: from wr-out-0506.google.com (wr-out-0506.google.com [64.233.184.232]) 13, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l47KVfnw019157 13, 28 -- for {microscopy-at-microscopy.com} ; Mon, 7 May 2007 15:31:42 -0500 13, 28 -- Received: by wr-out-0506.google.com with SMTP id i31so1832312wra 13, 28 -- for {microscopy-at-microscopy.com} ; Mon, 07 May 2007 13:31:41 -0700 (PDT) 13, 28 -- DKIM-Signature: a=rsa-sha1; c=relaxed/relaxed; 13, 28 -- d=gmail.com; s=beta; 13, 28 -- h=domainkey-signature:received:received:message-id:date:from:to:subject:in-reply-to:mime-version:content-type:content-transfer-encoding:content-disposition:references; 13, 28 -- b=q3RTmIkAb63uD7QpnHrl0NpdnuKFKdxQISpNF8kXIi39zfo7GHPWLAJ8MRte5aXwpJU4t2mLspOYLMW6qCwSwAZOViZTv3xro8YIrf2Ia8kRrV9zDEullZguN81LG68OL07O6swvtV0xqYBObIUdUcJ7LvjqmquOAdL8Dhx1mbo= 13, 28 -- DomainKey-Signature: a=rsa-sha1; c=nofws; 13, 28 -- d=gmail.com; s=beta; 13, 28 -- h=received:message-id:date:from:to:subject:in-reply-to:mime-version:content-type:content-transfer-encoding:content-disposition:references; 13, 28 -- b=DqU9GKYiC7aqQ6FAHzn0QP+yZTG8nWTVbC0zlxWEyXKemoJKBakM/IOEVSib3WEtwQyYNiqEq4f2TumXAAmQybfu3Vvg2vQjyGyM2P1nQ+CV//tutX6NPGrb35t7MF+e1eEzQi8e+CC3qQ08Li4CvxSzWLEaGEu08pomeS1tW8E= 13, 28 -- Received: by 10.114.167.2 with SMTP id p2mr947765wae.1178569900439; 13, 28 -- Mon, 07 May 2007 13:31:40 -0700 (PDT) 13, 28 -- Received: by 10.114.79.12 with HTTP; Mon, 7 May 2007 13:31:40 -0700 (PDT) 13, 28 -- Message-ID: {25e2b0d20705071331u3a9fc88dn811edd214693f8d0-at-mail.gmail.com} 13, 28 -- Date: Mon, 7 May 2007 16:31:40 -0400 13, 28 -- From: "Justin Kraft" {kraftpiano-at-gmail.com} 13, 28 -- To: microscopy-at-microscopy.com 13, 28 -- Subject: Re: [Microscopy] SEM: Intermittent Vacuum Leak 13, 28 -- In-Reply-To: {200705050135.l451ZYRx030376-at-ns.microscopy.com} 13, 28 -- MIME-Version: 1.0 13, 28 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 13, 28 -- Content-Transfer-Encoding: 7bit 13, 28 -- Content-Disposition: inline 13, 28 -- References: {200705050135.l451ZYRx030376-at-ns.microscopy.com} ==============================End of - Headers==============================
Just a quick not to say thank you to all of those who responded to my recent questions on the video capture from a Hitachi S4300 and the imaging of carrot cell walls.
Cheers
Colin Veitch
Electron Microscopist CSIRO Textile and Fibre Technology PO Box 21, BELMONT, Vic. 3216. Australia. colin.veitch-at-csiro.au http://www.tft.csiro.au
The information contained in this e-mail message may be privileged or confidential information. If you are not an intended recipient, you may not copy, distribute or take any action in reliance on it. If you have received this message in error, please telephone CSIRO Textile and Fibre Technology on +61 3 5246 4000.
==============================Original Headers============================== 8, 28 -- From Colin.Veitch-at-csiro.au Mon May 7 17:49:24 2007 8, 28 -- Received: from act-MTAout5.csiro.au (act-MTAout5.csiro.au [150.229.7.42]) 8, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l47MnNmH001485 8, 28 -- for {microscopy-at-msa.microscopy.com} ; Mon, 7 May 2007 17:49:24 -0500 8, 28 -- DomainKey-Signature: s=email; d=csiro.au; c=nofws; q=dns; b=PQjmuSnZw5fm6MW13O2lKys43qAJGwYkmJcFLXOpUjKRPrUziD3ZDdDcOxhqmjG0ATnJ+Diuoh117M+D9L1C3m+/soyJQmfvTRxqGCo7H8E9//h7uy+PCPe+gqoQOa/f; 8, 28 -- Received: from exgw1-mel.nexus.csiro.au ([138.194.3.56]) 8, 28 -- by act-ironport-int.csiro.au with ESMTP; 08 May 2007 08:52:43 +1000 8, 28 -- X-IronPort-AV: i="4.14,502,1170594000"; 8, 28 -- d="scan'208"; a="157024775:sNHT298823352" 8, 28 -- Received: from exvic5-gex.nexus.csiro.au ([138.194.194.6]) by exgw1-mel.nexus.csiro.au with Microsoft SMTPSVC(5.0.2195.6713); 8, 28 -- Tue, 8 May 2007 08:49:17 +1000 8, 28 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 8, 28 -- content-class: urn:content-classes:message 8, 28 -- MIME-Version: 1.0 8, 28 -- Content-Type: text/plain; 8, 28 -- charset="us-ascii" 8, 28 -- Subject: Many thanks for s4300 video question and imaging cell walls 8, 28 -- Date: Tue, 8 May 2007 08:49:17 +1000 8, 28 -- Message-ID: {32CDDDAA7161394599F00254949157492945EB-at-exvic5-gex.nexus.csiro.au} 8, 28 -- X-MS-Has-Attach: 8, 28 -- X-MS-TNEF-Correlator: 8, 28 -- Thread-Topic: Many thanks for s4300 video question and imaging cell walls 8, 28 -- Thread-Index: AceQ+fExIxrY/JVdTBOlDd0QO6GgBQ== 8, 28 -- From: {Colin.Veitch-at-csiro.au} 8, 28 -- To: {microscopy-at-msa.microscopy.com} 8, 28 -- X-OriginalArrivalTime: 07 May 2007 22:49:17.0785 (UTC) FILETIME=[F1979C90:01C790F9] 8, 28 -- Content-Transfer-Encoding: 8bit 8, 28 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l47MnNmH001485 ==============================End of - Headers==============================
I've recently had a problem with embedded blocks producing sections that look "muddy". The resin part of the section is fine, but the tissue is uniformly dark, as though it's embedded in mud, and is sometimes quite fragile, the section (tissue area) falling apart easily. Before I throw everything out and start with fresh reagents....any ideas? I receive the tissue already fixed, usually in glut/form in cacodylate buffer, not sure if it's from same operator each time; quality hasn't been good, but is adequate. It's then washed in cacodylate. Fixed in osmium (commercially prepared, clear and proven to have produced good results). Uranyl acetate block stain for an hour (same bottle I've been using for a long time). Ethanol and acetone dehydration (both kept dry with copper sulphate). Epon/ araldite embedding. The annoying thing is that it's not constant - one run will be fine, another ends up with the mud. I should do a run with different tissues, but don't have spare at the moment (and anyway I HATE embedding)....
It's probably something simple I'm doing wrong, but after all these years perhaps I'm blind to the obvious.
Cheers,
Diana
Diana van Driel Dept Ophthalmology Sydney University GPO Box 4337 Sydney, NSW AUSTRALIA 2001
When does the 'mud' appear? If it appears right after osmium tetroxide fixation it sounds like there might be some tannic acid in the tissue. I would ask the researcher for the complete protocol. I have seen this before when a researcher 'forgets' to tell me they read a paper that used tannic acid in the fix. If it appears after polymerization it sounds like incomplete dehydration, which will definitely lead to the fragile (I interpreted this as brittle) block. This is usually a result of a tissue piece that is too large for the solvent to penetrate all the way to the centre. Besides, what is to hate about embedding? The time consumed or the exposure to the nasty chemicals?
Good luck from Canada
Garnet
} } I've recently had a problem with embedded blocks producing sections } that look "muddy". The resin part of the section is fine, but the } tissue is uniformly dark, as though it's embedded in mud, and is } sometimes quite fragile, the section (tissue area) falling apart } easily. Before I throw everything out and start with fresh } reagents....any ideas? I receive the tissue already fixed, usually in } glut/form in cacodylate buffer, not sure if it's from same operator } each time; quality hasn't been good, but is adequate. It's then } washed in cacodylate. Fixed in osmium (commercially prepared, clear } and proven to have produced good results). Uranyl acetate block stain } for an hour (same bottle I've been using for a long time). Ethanol } and acetone dehydration (both kept dry with copper sulphate). Epon/ } araldite embedding. The annoying thing is that it's not constant - } one run will be fine, another ends up with the mud. I should do a run } with different tissues, but don't have spare at the moment (and } anyway I HATE embedding).... } } It's probably something simple I'm doing wrong, but after all these } years perhaps I'm blind to the obvious. } } Cheers, } } Diana }
-- Garnet Martens
Research Manager BioImaging Facility University of British Columbia 6270 University Blvd. Vancouver, B.C. Canada V6T 1Z4
phone 604-822-3354
==============================Original Headers============================== 8, 30 -- From gmartens-at-interchange.ubc.ca Tue May 8 10:16:11 2007 8, 30 -- Received: from mr3.mail-relay.ubc.ca (mr3.mail-relay.ubc.ca [137.82.45.5]) 8, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l48FG9sa011756 8, 30 -- for {Microscopy-at-microscopy.com} ; Tue, 8 May 2007 10:16:10 -0500 8, 30 -- Received: from mr3.mail-relay.ubc.ca (localhost [127.0.0.1]) 8, 30 -- by localhost (Postfix) with SMTP id 21650B65D 8, 30 -- for {Microscopy-at-microscopy.com} ; Tue, 8 May 2007 08:16:06 -0700 (PDT) 8, 30 -- Received: from mta2.interchange.ubc.ca (mta2.interchange.ubc.ca [142.103.145.70]) 8, 30 -- by mr3.mail-relay.ubc.ca (Postfix) with ESMTP 8, 30 -- for {Microscopy-at-microscopy.com} ; Tue, 8 May 2007 08:16:05 -0700 (PDT) 8, 30 -- Received: from [137.82.85.216] (0511.emlab.ubc.ca [137.82.85.216]) 8, 30 -- by smtp.interchange.ubc.ca 8, 30 -- (iPlanet Messaging Server 5.2 HotFix 1.21 (built Sep 8 2003)) 8, 30 -- with ESMTPA id {0JHQ005FCAESYM-at-smtp.interchange.ubc.ca} for 8, 30 -- Microscopy-at-microscopy.com; Tue, 08 May 2007 08:16:05 -0700 (PDT) 8, 30 -- Date: Tue, 08 May 2007 08:16:02 -0700 8, 30 -- From: Garnet Martens {gmartens-at-interchange.ubc.ca} 8, 30 -- Subject: Re: [Microscopy] muddy blocks 8, 30 -- In-reply-to: {200705080147.l481lRH9021761-at-ns.microscopy.com} 8, 30 -- To: dianavd-at-eye.usyd.edu.au 8, 30 -- Cc: Microscopy-at-microscopy.com 8, 30 -- Message-id: {a06240800c26642d81199-at-interchange.ubc.ca} 8, 30 -- MIME-version: 1.0 8, 30 -- Content-type: text/plain; format=flowed; charset=us-ascii 8, 30 -- References: {200705080147.l481lRH9021761-at-ns.microscopy.com} 8, 30 -- X-UBC-Scanned: Sophos PureMessage 5.3.1.294258, Antispam-Engine: 2.5.0.283055, Antispam-Data: 2007.5.8.80037 8, 30 -- X-UBC-Relayed: Relayed through mail-relay.ubc.ca 8, 30 -- X-PerlMx-Spam: Probability=7%, Report=__C230066_P5 0, __CT 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0 8, 30 -- X-Spam-Level: 8, 30 -- X-Spam-Flag: No ==============================End of - Headers==============================
Quorum Technologies has a variable size electrical carbon rod sharpener, please review the following URL for picture and description of the unit. http://www.quorumtech.com/Products/sc7605-carbon-rod-grinder.htm
Mike Dufraine EM-Product Manager Energy Beam Sciences,Inc.
dsherman-at-purdue.edu wrote:
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-- Michael F. Dufraine EM - Product Manager TEL 800-992-9037 X340 MDufraine-at-ebsciences.com www.ebsciences.com
==============================Original Headers============================== 7, 27 -- From mdufraine-at-ebsciences.com Tue May 8 10:19:18 2007 7, 27 -- Received: from mx-outbound01.easydns.com (mailout.easydns.com [205.210.42.54]) 7, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l48FJIAI014315 7, 27 -- for {microscopy-at-microscopy.com} ; Tue, 8 May 2007 10:19:18 -0500 7, 27 -- Received: from mail.ebsciences.com (unknown [69.182.224.2]) 7, 27 -- (using TLSv1 with cipher DHE-RSA-AES256-SHA (256/256 bits)) 7, 27 -- (No client certificate requested) 7, 27 -- by mx-outbound01.easydns.com (Postfix) with ESMTP id DE6CB7F73; 7, 27 -- Tue, 8 May 2007 11:19:16 -0400 (EDT) 7, 27 -- Received: from mdufraine.ebsciences.private ([10.10.0.196]) 7, 27 -- by mail.ebsciences.com with esmtpsa (TLSv1:AES256-SHA:256) 7, 27 -- (Exim 4.62) 7, 27 -- (envelope-from {mdufraine-at-ebsciences.com} ) 7, 27 -- id 1HlRTA-00041x-Kt; Tue, 08 May 2007 11:19:16 -0400 7, 27 -- Message-ID: {464094F2.4090301-at-ebsciences.com} 7, 27 -- Date: Tue, 08 May 2007 11:19:14 -0400 7, 27 -- From: Mike Dufraine {mdufraine-at-ebsciences.com} 7, 27 -- Organization: Energy Beam Sciences 7, 27 -- User-Agent: Mozilla Thunderbird 1.0.2 (Windows/20050317) 7, 27 -- X-Accept-Language: en-us, en 7, 27 -- MIME-Version: 1.0 7, 27 -- To: dsherman-at-purdue.edu, microscopy-at-microscopy.com 7, 27 -- Subject: Re: [Microscopy] Carbon rod sharpener 7, 27 -- References: {200705041923.l44JN9Tp000563-at-ns.microscopy.com} 7, 27 -- In-Reply-To: {200705041923.l44JN9Tp000563-at-ns.microscopy.com} 7, 27 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 7, 27 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
I'd like to get some input on sample preparation with respect to silicon.
PROBLEM:
I have an FEI XL-50 FESEM that was originally designed to accept flat samples, essentially silicon wafers. It does not have what one would consider a typical exchange port. The exchange port is robotically controlled, and the maximum sample height, including stub, must be 5 mm or less. I must view these samples on edge, i.e. 90 degrees. This presents a problem in that I must cleave two parallel sections very close to each other. Perhaps diamond cutters can do this, but my hands are too shaky.
QUESTION:
Is there a device, or method, that would allow me to make these cleaves ~ 8 mm apart with any reasonable control? I found stubs that are low profile, 38 and 90 degrees, from the nice people at Ted Pella, but I still have this issue of doing two parallel cleaves very close together.
Just for info. purposes I am in the silicon MEMS development arena.
If you feel your reply is of general interest to this community, please reply to all, or you may contact me directly.
Regards to all in this small world,
Peter Tomic Renaissance Wireless Corp.
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==============================Original Headers============================== 13, 25 -- From peter.tomic-at-renwireless.com Tue May 8 12:33:06 2007 13, 25 -- Received: from mail.rw.local (4.piok2.xdsl.nauticom.net [209.195.176.101]) 13, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l48HX6om005343 13, 25 -- for {Microscopy-at-microscopy.com} ; Tue, 8 May 2007 12:33:06 -0500 13, 25 -- Received: from localhost (localhost [127.0.0.1]) 13, 25 -- by mail.rw.local (Postfix) with ESMTP id 97112241A0 13, 25 -- for {Microscopy-at-microscopy.com} ; Tue, 8 May 2007 13:33:05 -0400 (EDT) 13, 25 -- Received: from mail.rw.local ([127.0.0.1]) 13, 25 -- by localhost (mail.rw.local [127.0.0.1]) (amavisd-new, port 10024) with ESMTP 13, 25 -- id 30832-08 for {Microscopy-at-microscopy.com} ; 13, 25 -- Tue, 8 May 2007 13:33:05 -0400 (EDT) 13, 25 -- Received: from [127.0.0.1] (unknown [10.11.10.142]) 13, 25 -- by mail.rw.local (Postfix) with ESMTP id 2E11024168 13, 25 -- for {Microscopy-at-microscopy.com} ; Tue, 8 May 2007 13:33:05 -0400 (EDT) 13, 25 -- Message-ID: {4640B45E.6040307-at-renwireless.com} 13, 25 -- Date: Tue, 08 May 2007 13:33:18 -0400 13, 25 -- From: Peter Tomic {peter.tomic-at-renwireless.com} 13, 25 -- Organization: Renaissance Wireless Corp. 13, 25 -- User-Agent: Thunderbird 1.5.0.10 (Windows/20070221) 13, 25 -- MIME-Version: 1.0 13, 25 -- To: Microscopy-at-microscopy.com 13, 25 -- Subject: Silicon Cross-section sample preparation 13, 25 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 13, 25 -- Content-Transfer-Encoding: 7bit 13, 25 -- X-Virus-Scanned: amavisd-new at renwireless.com ==============================End of - Headers==============================
I am trying to find resolution of a problem with our CM300 TEM with a LaB6 cathode.
When trying to get diffraction from an almost amorphous material I have noticed a presence of relatively weak but clearly visible perfectly circular ring of intensity around the central transmitted spot. The ring is not centered around the transmitted spot. Its position varies when operating the beam shift controls, its size varies with the C2 aperture size when in diffraction mode. It is also present when there is no sample under the beam. When there is strongly reflecting crystalline material the ring is almost invisible due to its weak relative intensity. It is present at any accelerating voltage.
I am trying to get some ideas about what might be causing the ring and how to eliminate it.
The ring is also visible when in LM imaging mode (image is formed by the diffraction lens) and when the beam is focused to a spot. In LM mode the ring has strange shape. It has sharp circular outline on its outer edge and it has irregular shape and outline on its inner edge.
Using the free lens control option I have made the following observations:
In LM imaging mode with beam focused to a spot. When changing the C1 lens current -the ring changes size and focus but does not rotate. changing C2 lens current - change in focus only, plus some rotation changing Twin lens current - change of size and focus, no rotation changing Objective lens current - change in size and focus, some rotation changing Diffraction lens current - change in size and focus, no rotation changing Intermediate lens current - change in size and focus, no rotation changing P1 lens current - change in size and focus, some rotation changing P2 lens current - rotation only
In diffraction mode with fully spread beam: position varies when operating the beam shift controls size varies with the C2 aperture size.
FEI support engineers have not been succesful in identifing the problem so far. It was suggested that it is due to undersaturated LaB6 cathode, a possibility which we have clearly eliminated as a possible source.
Thanks for any hints our suggestions.
Krassimir. _______________________________________ Krassimir N. Bozhilov Central Facility for Advanced Microscopy and Microanalysis University of California Riverside, CA 92521
tel 951 827 2998 fax 951 827 2489 bozhilov-at-ucr.edu _______________________________________
==============================Original Headers============================== 14, 19 -- From bozhilov-at-ucr.edu Tue May 8 13:47:05 2007 14, 19 -- Received: from sentoku.ucr.edu (sentoku.ucr.edu [138.23.226.244]) 14, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l48Il5CQ018432 14, 19 -- for {microscopy-at-microscopy.com} ; Tue, 8 May 2007 13:47:05 -0500 14, 19 -- Received: from [138.23.185.162] (igppb116g.ucr.edu [138.23.185.162]) 14, 19 -- by sentoku.ucr.edu (MOS 3.6.6-GR) 14, 19 -- with ESMTP id BJQ30585 (AUTH via LOGINBEFORESMTP) 14, 19 -- for {microscopy-at-microscopy.com} ; 14, 19 -- Tue, 8 May 2007 11:47:04 -0700 (PDT) 14, 19 -- Mime-Version: 1.0 (Apple Message framework v752.2) 14, 19 -- Content-Transfer-Encoding: 7bit 14, 19 -- Message-Id: {102C9F08-32B8-4A7D-9620-81C1F05F2CE1-at-ucr.edu} 14, 19 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 14, 19 -- To: microscopy-at-microscopy.com 14, 19 -- From: KN Bozhilov {bozhilov-at-ucr.edu} 14, 19 -- Subject: Diffracation ring problem 14, 19 -- Date: Tue, 8 May 2007 11:47:03 -0700 14, 19 -- X-Mailer: Apple Mail (2.752.2) 14, 19 -- X-Junkmail-Whitelist: YES (by domain whitelist at sentoku.ucr.edu) ==============================End of - Headers==============================
Consider the possibility that it might be visible light from the hot cathode coming down the column, i.e. like a flashlight.
This argument is abetted by the sharp outside edge, from apertures along the way, and a diffuse inner edge and the fact that you see it without a specimen loaded. However, it isn't immediately obvious how or why it changes size and shape with all of your other perturbations.
Can you partly eclipse it by moving apertures around?
Ron Anderson
bozhilov-at-ucr.edu wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Hi all, } } I am trying to find resolution of a problem with our CM300 TEM with a } LaB6 cathode. } } When trying to get diffraction from an almost amorphous material I } have noticed a presence of relatively weak but clearly visible } perfectly circular ring of intensity around the central transmitted } spot. The ring is not centered around the transmitted spot. Its } position varies when operating the beam shift controls, its size } varies with the C2 aperture size when in diffraction mode. It is also } present when there is no sample under the beam. When there is } strongly reflecting crystalline material the ring is almost invisible } due to its weak relative intensity. It is present at any accelerating } voltage. } } I am trying to get some ideas about what might be causing the ring } and how to eliminate it. } } The ring is also visible when in LM imaging mode (image is formed by } the diffraction lens) } and when the beam is focused to a spot. In LM mode the ring has } strange shape. It has sharp circular outline on its outer edge and it } has irregular shape and outline on its inner edge. } } Using the free lens control option I have made the following } observations: } } In LM imaging mode with beam focused to a spot. } When changing the C1 lens current -the ring changes size and focus } but does not rotate. } changing C2 lens current - change in focus only, plus some rotation } changing Twin lens current - change of size and focus, no rotation } changing Objective lens current - change in size and focus, some } rotation } changing Diffraction lens current - change in size and focus, no } rotation } changing Intermediate lens current - change in size and focus, no } rotation } changing P1 lens current - change in size and focus, some rotation } changing P2 lens current - rotation only } } In diffraction mode with fully spread beam: } position varies when operating the beam shift controls } size varies with the C2 aperture size. } } FEI support engineers have not been succesful in identifing the } problem so far. It was suggested that it is due to undersaturated } LaB6 cathode, a possibility which we have clearly eliminated as a } possible source. } } Thanks for any hints our suggestions. } } Krassimir. } _______________________________________ } Krassimir N. Bozhilov } Central Facility for Advanced Microscopy and Microanalysis } University of California } Riverside, CA 92521 } } tel 951 827 2998 } fax 951 827 2489 } bozhilov-at-ucr.edu } _______________________________________ } } } } ==============================Original Headers============================== } 14, 19 -- From bozhilov-at-ucr.edu Tue May 8 13:47:05 2007 } 14, 19 -- Received: from sentoku.ucr.edu (sentoku.ucr.edu [138.23.226.244]) } 14, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l48Il5CQ018432 } 14, 19 -- for {microscopy-at-microscopy.com} ; Tue, 8 May 2007 13:47:05 -0500 } 14, 19 -- Received: from [138.23.185.162] (igppb116g.ucr.edu [138.23.185.162]) } 14, 19 -- by sentoku.ucr.edu (MOS 3.6.6-GR) } 14, 19 -- with ESMTP id BJQ30585 (AUTH via LOGINBEFORESMTP) } 14, 19 -- for {microscopy-at-microscopy.com} ; } 14, 19 -- Tue, 8 May 2007 11:47:04 -0700 (PDT) } 14, 19 -- Mime-Version: 1.0 (Apple Message framework v752.2) } 14, 19 -- Content-Transfer-Encoding: 7bit } 14, 19 -- Message-Id: {102C9F08-32B8-4A7D-9620-81C1F05F2CE1-at-ucr.edu} } 14, 19 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed } 14, 19 -- To: microscopy-at-microscopy.com } 14, 19 -- From: KN Bozhilov {bozhilov-at-ucr.edu} } 14, 19 -- Subject: Diffracation ring problem } 14, 19 -- Date: Tue, 8 May 2007 11:47:03 -0700 } 14, 19 -- X-Mailer: Apple Mail (2.752.2) } 14, 19 -- X-Junkmail-Whitelist: YES (by domain whitelist at sentoku.ucr.edu) } ==============================End of - Headers============================== } } }
==============================Original Headers============================== 6, 19 -- From randerson20-at-tampabay.rr.com Tue May 8 14:44:37 2007 6, 19 -- Received: from ms-smtp-01.tampabay.rr.com (ms-smtp-01.tampabay.rr.com [65.32.5.131]) 6, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l48JibIA031194 6, 19 -- for {Microscopy-at-Microscopy.Com} ; Tue, 8 May 2007 14:44:37 -0500 6, 19 -- Received: from [127.0.0.1] (rrcs-24-73-73-214.se.biz.rr.com [24.73.73.214]) 6, 19 -- by ms-smtp-01.tampabay.rr.com (8.13.6/8.13.6) with ESMTP id l48JiYet014135; 6, 19 -- Tue, 8 May 2007 15:44:36 -0400 (EDT) 6, 19 -- Message-ID: {4640D320.4010203-at-tampabay.rr.com} 6, 19 -- Date: Tue, 08 May 2007 15:44:32 -0400 6, 19 -- From: Ronald Anderson {randerson20-at-tampabay.rr.com} 6, 19 -- User-Agent: Thunderbird 1.5.0.10 (Windows/20070221) 6, 19 -- MIME-Version: 1.0 6, 19 -- To: bozhilov-at-ucr.edu, Listserver {Microscopy-at-Microscopy.Com} 6, 19 -- Subject: Re: [Microscopy] Diffracation ring problem 6, 19 -- References: {200705081847.l48IlJhr018618-at-ns.microscopy.com} 6, 19 -- In-Reply-To: {200705081847.l48IlJhr018618-at-ns.microscopy.com} 6, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 6, 19 -- Content-Transfer-Encoding: 7bit 6, 19 -- X-Virus-Scanned: Symantec AntiVirus Scan Engine ==============================End of - Headers==============================
When the beam is focused to a probe in LM mode moving the C2 or the diffrcation aperture does not eclipse the ring. It chages only the intensity of the light or when moved too far blocks all the light. Only by introducing and moving the objective aperture one can select/ block part of the ring, respectively the central spot.
Krassimir.
On May 8, 2007, at 12:46 PM, randerson20-at-tampabay.rr.com wrote:
} } } } ---------------------------------------------------------------------- } ------ } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } Consider the possibility that it might be visible light from the hot } cathode coming down the column, i.e. like a flashlight. } } This argument is abetted by the sharp outside edge, from apertures } along } the way, and a diffuse inner edge and the fact that you see it } without a } specimen loaded. However, it isn't immediately obvious how or why it } changes size and shape with all of your other perturbations. } } Can you partly eclipse it by moving apertures around? } } Ron Anderson } } bozhilov-at-ucr.edu wrote: } } --------------------------------------------------------------------- } } ------- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } } America } } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } } MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } --------------------------------------------------------------------- } } ------- } } } } Hi all, } } } } I am trying to find resolution of a problem with our CM300 TEM with a } } LaB6 cathode. } } } } When trying to get diffraction from an almost amorphous material I } } have noticed a presence of relatively weak but clearly visible } } perfectly circular ring of intensity around the central transmitted } } spot. The ring is not centered around the transmitted spot. Its } } position varies when operating the beam shift controls, its size } } varies with the C2 aperture size when in diffraction mode. It is also } } present when there is no sample under the beam. When there is } } strongly reflecting crystalline material the ring is almost invisible } } due to its weak relative intensity. It is present at any accelerating } } voltage. } } } } I am trying to get some ideas about what might be causing the ring } } and how to eliminate it. } } } } The ring is also visible when in LM imaging mode (image is formed by } } the diffraction lens) } } and when the beam is focused to a spot. In LM mode the ring has } } strange shape. It has sharp circular outline on its outer edge and it } } has irregular shape and outline on its inner edge. } } } } Using the free lens control option I have made the following } } observations: } } } } In LM imaging mode with beam focused to a spot. } } When changing the C1 lens current -the ring changes size and focus } } but does not rotate. } } changing C2 lens current - change in focus only, plus some rotation } } changing Twin lens current - change of size and focus, no rotation } } changing Objective lens current - change in size and focus, some } } rotation } } changing Diffraction lens current - change in size and focus, no } } rotation } } changing Intermediate lens current - change in size and focus, no } } rotation } } changing P1 lens current - change in size and focus, some rotation } } changing P2 lens current - rotation only } } } } In diffraction mode with fully spread beam: } } position varies when operating the beam shift controls } } size varies with the C2 aperture size. } } } } FEI support engineers have not been succesful in identifing the } } problem so far. It was suggested that it is due to undersaturated } } LaB6 cathode, a possibility which we have clearly eliminated as a } } possible source. } } } } Thanks for any hints our suggestions. } } } } Krassimir. } } _______________________________________ } } Krassimir N. Bozhilov } } Central Facility for Advanced Microscopy and Microanalysis } } University of California } } Riverside, CA 92521 } } } } tel 951 827 2998 } } fax 951 827 2489 } } bozhilov-at-ucr.edu } } _______________________________________ } } } } } } } } ==============================Original } } Headers============================== } } 14, 19 -- From bozhilov-at-ucr.edu Tue May 8 13:47:05 2007 } } 14, 19 -- Received: from sentoku.ucr.edu (sentoku.ucr.edu } } [138.23.226.244]) } } 14, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } } ESMTP id l48Il5CQ018432 } } 14, 19 -- for {microscopy-at-microscopy.com} ; Tue, 8 May 2007 } } 13:47:05 -0500 } } 14, 19 -- Received: from [138.23.185.162] (igppb116g.ucr.edu } } [138.23.185.162]) } } 14, 19 -- by sentoku.ucr.edu (MOS 3.6.6-GR) } } 14, 19 -- with ESMTP id BJQ30585 (AUTH via LOGINBEFORESMTP) } } 14, 19 -- for {microscopy-at-microscopy.com} ; } } 14, 19 -- Tue, 8 May 2007 11:47:04 -0700 (PDT) } } 14, 19 -- Mime-Version: 1.0 (Apple Message framework v752.2) } } 14, 19 -- Content-Transfer-Encoding: 7bit } } 14, 19 -- Message-Id: {102C9F08-32B8-4A7D-9620-81C1F05F2CE1-at-ucr.edu} } } 14, 19 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; } } format=flowed } } 14, 19 -- To: microscopy-at-microscopy.com } } 14, 19 -- From: KN Bozhilov {bozhilov-at-ucr.edu} } } 14, 19 -- Subject: Diffracation ring problem } } 14, 19 -- Date: Tue, 8 May 2007 11:47:03 -0700 } } 14, 19 -- X-Mailer: Apple Mail (2.752.2) } } 14, 19 -- X-Junkmail-Whitelist: YES (by domain whitelist at } } sentoku.ucr.edu) } } ==============================End of - } } Headers============================== } } } } } } } } } ==============================Original } Headers============================== } 6, 19 -- From randerson20-at-tampabay.rr.com Tue May 8 14:44:37 2007 } 6, 19 -- Received: from ms-smtp-01.tampabay.rr.com (ms- } smtp-01.tampabay.rr.com [65.32.5.131]) } 6, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP } id l48JibIA031194 } 6, 19 -- for {Microscopy-at-Microscopy.Com} ; Tue, 8 May 2007 14:44:37 } -0500 } 6, 19 -- Received: from [127.0.0.1] } (rrcs-24-73-73-214.se.biz.rr.com [24.73.73.214]) } 6, 19 -- by ms-smtp-01.tampabay.rr.com (8.13.6/8.13.6) with ESMTP } id l48JiYet014135; } 6, 19 -- Tue, 8 May 2007 15:44:36 -0400 (EDT) } 6, 19 -- Message-ID: {4640D320.4010203-at-tampabay.rr.com} } 6, 19 -- Date: Tue, 08 May 2007 15:44:32 -0400 } 6, 19 -- From: Ronald Anderson {randerson20-at-tampabay.rr.com} } 6, 19 -- User-Agent: Thunderbird 1.5.0.10 (Windows/20070221) } 6, 19 -- MIME-Version: 1.0 } 6, 19 -- To: bozhilov-at-ucr.edu, Listserver {Microscopy-at-Microscopy.Com} } 6, 19 -- Subject: Re: [Microscopy] Diffracation ring problem } 6, 19 -- References: {200705081847.l48IlJhr018618-at-ns.microscopy.com} } 6, 19 -- In-Reply-To: {200705081847.l48IlJhr018618-at-ns.microscopy.com} } 6, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed } 6, 19 -- Content-Transfer-Encoding: 7bit } 6, 19 -- X-Virus-Scanned: Symantec AntiVirus Scan Engine } ==============================End of - } Headers==============================
==============================Original Headers============================== 6, 21 -- From bozhilov-at-ucr.edu Tue May 8 16:19:34 2007 6, 21 -- Received: from sentry.ucr.edu (sentry.ucr.edu [138.23.226.224]) 6, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l48LJYuO011829 6, 21 -- for {microscopy-at-microscopy.com} ; Tue, 8 May 2007 16:19:34 -0500 6, 21 -- Received: from [138.23.185.162] (igppb116g.ucr.edu [138.23.185.162]) 6, 21 -- by sentry.ucr.edu (MOS 3.6.6-GR) 6, 21 -- with ESMTP id ERG46680 (AUTH via LOGINBEFORESMTP); 6, 21 -- Tue, 8 May 2007 14:19:30 -0700 (PDT) 6, 21 -- In-Reply-To: {200705081946.l48JkjKd002181-at-ns.microscopy.com} 6, 21 -- References: {200705081946.l48JkjKd002181-at-ns.microscopy.com} 6, 21 -- Mime-Version: 1.0 (Apple Message framework v752.2) 6, 21 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 6, 21 -- Message-Id: {13EA274E-9182-48C9-9AAB-F00506AD735D-at-ucr.edu} 6, 21 -- Cc: microscopy-at-microscopy.com 6, 21 -- Content-Transfer-Encoding: 7bit 6, 21 -- From: KN Bozhilov {bozhilov-at-ucr.edu} 6, 21 -- Subject: Re: [Microscopy] Re: Diffracation ring problem 6, 21 -- Date: Tue, 8 May 2007 14:19:30 -0700 6, 21 -- To: randerson20-at-tampabay.rr.com 6, 21 -- X-Mailer: Apple Mail (2.752.2) 6, 21 -- X-Junkmail-Whitelist: YES (by domain whitelist at sentry.ucr.edu) ==============================End of - Headers==============================
Peter, It's much easier to control a cleave in silicon the thinner the sample. Back thin your sample using a lapping fixture to keep the sample parallel faced. Then simply use a fine scribe to start and propagate your cleave.
Another option for you is to use the Tripod Polisher(R) for your cross section. You could easily polish your samples to a specific site and control the thickness of them very accurately. It would take you a little longer than simply cleaving, but if you have to back thin your samples, it would be a wash. If you need any literature on the TP technique, please contact me offline.
Disclaimer: South Bay Technology manufactures and sells the Tripod Polisher(R).
-Scott
Scott D. Walck, Ph.D. Technical Director South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
US Toll Free: 1-800-728-2233 Tel: (949) 492-2600 Fax: (949) 492-1499
www.southbaytech.com walck-at-southbaytech.com
-----Original Message----- X-from: peter.tomic-at-renwireless.com [mailto:peter.tomic-at-renwireless.com] Sent: Tuesday, May 08, 2007 10:37 AM To: Walck-at-SouthBayTech.com
Microscopy Folks,
I'd like to get some input on sample preparation with respect to silicon.
PROBLEM:
I have an FEI XL-50 FESEM that was originally designed to accept flat samples, essentially silicon wafers. It does not have what one would consider a typical exchange port. The exchange port is robotically controlled, and the maximum sample height, including stub, must be 5 mm or less. I must view these samples on edge, i.e. 90 degrees. This presents a problem in that I must cleave two parallel sections very close to each other. Perhaps diamond cutters can do this, but my hands are too shaky.
QUESTION:
Is there a device, or method, that would allow me to make these cleaves ~ 8 mm apart with any reasonable control? I found stubs that are low profile, 38 and 90 degrees, from the nice people at Ted Pella, but I still have this issue of doing two parallel cleaves very close together.
Just for info. purposes I am in the silicon MEMS development arena.
If you feel your reply is of general interest to this community, please reply to all, or you may contact me directly.
Regards to all in this small world,
Peter Tomic Renaissance Wireless Corp.
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==============================Original Headers============================== 13, 25 -- From peter.tomic-at-renwireless.com Tue May 8 12:33:06 2007 13, 25 -- Received: from mail.rw.local (4.piok2.xdsl.nauticom.net [209.195.176.101]) 13, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l48HX6om005343 13, 25 -- for {Microscopy-at-microscopy.com} ; Tue, 8 May 2007 12:33:06 -0500 13, 25 -- Received: from localhost (localhost [127.0.0.1]) 13, 25 -- by mail.rw.local (Postfix) with ESMTP id 97112241A0 13, 25 -- for {Microscopy-at-microscopy.com} ; Tue, 8 May 2007 13:33:05 -0400 (EDT) 13, 25 -- Received: from mail.rw.local ([127.0.0.1]) 13, 25 -- by localhost (mail.rw.local [127.0.0.1]) (amavisd-new, port 10024) with ESMTP 13, 25 -- id 30832-08 for {Microscopy-at-microscopy.com} ; 13, 25 -- Tue, 8 May 2007 13:33:05 -0400 (EDT) 13, 25 -- Received: from [127.0.0.1] (unknown [10.11.10.142]) 13, 25 -- by mail.rw.local (Postfix) with ESMTP id 2E11024168 13, 25 -- for {Microscopy-at-microscopy.com} ; Tue, 8 May 2007 13:33:05 -0400 (EDT) 13, 25 -- Message-ID: {4640B45E.6040307-at-renwireless.com} 13, 25 -- Date: Tue, 08 May 2007 13:33:18 -0400 13, 25 -- From: Peter Tomic {peter.tomic-at-renwireless.com} 13, 25 -- Organization: Renaissance Wireless Corp. 13, 25 -- User-Agent: Thunderbird 1.5.0.10 (Windows/20070221) 13, 25 -- MIME-Version: 1.0 13, 25 -- To: Microscopy-at-microscopy.com 13, 25 -- Subject: Silicon Cross-section sample preparation 13, 25 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 13, 25 -- Content-Transfer-Encoding: 7bit 13, 25 -- X-Virus-Scanned: amavisd-new at renwireless.com ==============================End of - Headers==============================
==============================Original Headers============================== 24, 22 -- From walck-at-southbaytech.com Tue May 8 16:51:28 2007 24, 22 -- Received: from flpvm23.prodigy.net (flpvm23.prodigy.net [207.115.20.53]) 24, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l48LpRLZ024095 24, 22 -- for {Microscopy-at-microscopy.com} ; Tue, 8 May 2007 16:51:28 -0500 24, 22 -- X-ORBL: [64.169.217.123] 24, 22 -- Received: from dynamicbl8uno3 (adsl-64-169-217-123.dsl.lsan03.pacbell.net [64.169.217.123]) 24, 22 -- by flpvm23.prodigy.net (8.13.8 out.dk.spool/8.13.8) with ESMTP id l48LpUjP007984; 24, 22 -- Tue, 8 May 2007 14:51:30 -0700 24, 22 -- From: "Scott Walck" {walck-at-southbaytech.com} 24, 22 -- To: {peter.tomic-at-renwireless.com} 24, 22 -- Cc: {Microscopy-at-microscopy.com} 24, 22 -- Subject: RE: [Microscopy] Silicon Cross-section sample preparation 24, 22 -- Date: Tue, 8 May 2007 14:51:51 -0700 24, 22 -- Message-ID: {007601c791bb$163c7040$7801a8c0-at-dynamicbl8uno3} 24, 22 -- MIME-Version: 1.0 24, 22 -- Content-Type: text/plain; 24, 22 -- charset="us-ascii" 24, 22 -- Content-Transfer-Encoding: 7bit 24, 22 -- X-Mailer: Microsoft Office Outlook 11 24, 22 -- In-Reply-To: {200705081736.l48HauXU011121-at-ns.microscopy.com} 24, 22 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 24, 22 -- Thread-Index: AceRl4Aub45e4dtXTx+x2jJTNjpLbAAIU49A ==============================End of - Headers==============================
At the request of an investigator who was interested in plant microtubules, we fixed arabidopsis hypocotyles in:
2.5% formaldehyde, 2% glutaraldehyde, in PEMT (100mM pipes-KOH, pH 6.9, 5mM EGTA, 2 mM MgCl2, 0.05% Triton X-100) for 1h at RT
This was followed by standard 2% OsO4, ETOH dehydration and embedding in Spurr's resin.
The results were less than pleasing. The membranes seemed soft with little crisp clarity anywhere. In contrast we fixed maize leaves using:
2.5% glut + 2% PAF in 0.1M cacodylate, pH 6.8, as the primary fix with the rest the same as above and got great membranes, sharp chloroplast granae stacks, etc.
Only real difference was the buffer. Have any of you used a similar pipes buffer and do you have any comments/ideas of why the one prep was good and the other not?
Thanks, Debby
Debby Sherman, Director Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www.agriculture.purdue.edu/microscopy
==============================Original Headers============================== 11, 22 -- From dsherman-at-purdue.edu Tue May 8 17:21:46 2007 11, 22 -- Received: from 1061exfe01.adpc.purdue.edu (1061exfe01.adpc.purdue.edu [128.210.63.221]) 11, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l48MLkbb003574 11, 22 -- for {microscopy-at-microscopy.com} ; Tue, 8 May 2007 17:21:46 -0500 11, 22 -- Received: from exchange.purdue.edu ([172.21.6.24]) by 1061exfe01.adpc.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 11, 22 -- Tue, 8 May 2007 18:21:46 -0400 11, 22 -- Received: from 74.140.109.174 ([74.140.109.174]) by EXCH04.purdue.lcl ([172.21.6.26]) via Exchange Front-End Server exch.itap.purdue.edu ([128.210.63.104]) with Microsoft Exchange Server HTTP-DAV ; 11, 22 -- Tue, 8 May 2007 22:21:46 +0000 11, 22 -- User-Agent: Microsoft-Entourage/11.3.3.061214 11, 22 -- Date: Tue, 08 May 2007 18:21:46 -0400 11, 22 -- Subject: Plant fixation-pipes 11, 22 -- From: Debby Sherman {dsherman-at-purdue.edu} 11, 22 -- To: "message to: MSA list" {microscopy-at-microscopy.com} 11, 22 -- Message-ID: {C266703A.CCEF%dsherman-at-purdue.edu} 11, 22 -- Thread-Topic: Plant fixation-pipes 11, 22 -- Thread-Index: AceRv0N2gfJrsf2yEdutAAAKlcoUxg== 11, 22 -- Mime-version: 1.0 11, 22 -- Content-type: text/plain; 11, 22 -- charset="ISO-8859-1" 11, 22 -- X-OriginalArrivalTime: 08 May 2007 22:21:46.0692 (UTC) FILETIME=[43E09040:01C791BF] 11, 22 -- Content-Transfer-Encoding: 8bit 11, 22 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l48MLkbb003574 ==============================End of - Headers==============================
Debby, The first thing that I saw was Triton X-100! I don't imagine that including a surfactant with your primary fixative would be any good for the membranes. I have used straight PIPES (without the additives)in plant tissues with good results (but not better than with cacodylate or phosphate buffers). Kim
dsherman-at-purdue.edu wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Hi all, } } At the request of an investigator who was interested in plant microtubules, } we fixed arabidopsis hypocotyles in: } } 2.5% formaldehyde, 2% glutaraldehyde, in PEMT (100mM pipes-KOH, pH } 6.9, 5mM EGTA, 2 mM MgCl2, 0.05% Triton X-100) for 1h at RT } } This was followed by standard 2% OsO4, ETOH dehydration and embedding in } Spurr's resin. } } The results were less than pleasing. The membranes seemed soft with little } crisp clarity anywhere. In contrast we fixed maize leaves using: } } 2.5% glut + 2% PAF in 0.1M cacodylate, pH 6.8, as the primary fix with the } rest the same as above and got great membranes, sharp chloroplast granae } stacks, etc. } } Only real difference was the buffer. Have any of you used a similar pipes } buffer and do you have any comments/ideas of why the one prep was good and } the other not? } } Thanks, } Debby } } Debby Sherman, Director Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } S-052 Whistler Building } 170 S. University Street } West Lafayette, IN 47907 } http://www.agriculture.purdue.edu/microscopy } } } } ==============================Original Headers============================== } 11, 22 -- From dsherman-at-purdue.edu Tue May 8 17:21:46 2007 } 11, 22 -- Received: from 1061exfe01.adpc.purdue.edu (1061exfe01.adpc.purdue.edu [128.210.63.221]) } 11, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l48MLkbb003574 } 11, 22 -- for {microscopy-at-microscopy.com} ; Tue, 8 May 2007 17:21:46 -0500 } 11, 22 -- Received: from exchange.purdue.edu ([172.21.6.24]) by 1061exfe01.adpc.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); } 11, 22 -- Tue, 8 May 2007 18:21:46 -0400 } 11, 22 -- Received: from 74.140.109.174 ([74.140.109.174]) by EXCH04.purdue.lcl ([172.21.6.26]) via Exchange Front-End Server exch.itap.purdue.edu ([128.210.63.104]) with Microsoft Exchange Server HTTP-DAV ; } 11, 22 -- Tue, 8 May 2007 22:21:46 +0000 } 11, 22 -- User-Agent: Microsoft-Entourage/11.3.3.061214 } 11, 22 -- Date: Tue, 08 May 2007 18:21:46 -0400 } 11, 22 -- Subject: Plant fixation-pipes } 11, 22 -- From: Debby Sherman {dsherman-at-purdue.edu} } 11, 22 -- To: "message to: MSA list" {microscopy-at-microscopy.com} } 11, 22 -- Message-ID: {C266703A.CCEF%dsherman-at-purdue.edu} } 11, 22 -- Thread-Topic: Plant fixation-pipes } 11, 22 -- Thread-Index: AceRv0N2gfJrsf2yEdutAAAKlcoUxg== } 11, 22 -- Mime-version: 1.0 } 11, 22 -- Content-type: text/plain; } 11, 22 -- charset="ISO-8859-1" } 11, 22 -- X-OriginalArrivalTime: 08 May 2007 22:21:46.0692 (UTC) FILETIME=[43E09040:01C791BF] } 11, 22 -- Content-Transfer-Encoding: 8bit } 11, 22 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l48MLkbb003574 } ==============================End of - Headers============================== } }
-- Kim Rensing Ph.D. Assistant Research Professor Manager, Microscopy and Imaging Facility
University of Calgary Health Sciences Centre B129 - 3330 Hospital Drive NW Calgary, AB, Canada T2N 4N1 403-220-3488 krensing-at-ucalgary.ca
==============================Original Headers============================== 4, 22 -- From krensing-at-ucalgary.ca Tue May 8 17:40:14 2007 4, 22 -- Received: from smtp2.ucalgary.ca (smtp2.ucalgary.ca [136.159.34.223]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l48MeDYD015305 4, 22 -- for {microscopy-at-microscopy.com} ; Tue, 8 May 2007 17:40:13 -0500 4, 22 -- Received: from [136.159.164.163] (unknown [136.159.164.163]) 4, 22 -- by smtp2.ucalgary.ca (Postfix) with ESMTP id 6063210009 4, 22 -- for {microscopy-at-microscopy.com} ; Tue, 8 May 2007 16:40:09 -0600 (MDT) 4, 22 -- Message-ID: {4640FC42.8060402-at-ucalgary.ca} 4, 22 -- Date: Tue, 08 May 2007 16:40:02 -0600 4, 22 -- From: Kim Rensing {krensing-at-ucalgary.ca} 4, 22 -- Organization: Microscopy and Imaging Facility, U. of Calgary 4, 22 -- User-Agent: Thunderbird 1.5.0.10 (Windows/20070221) 4, 22 -- MIME-Version: 1.0 4, 22 -- To: microscopy-at-microscopy.com 4, 22 -- Subject: Re: [Microscopy] Plant fixation-pipes 4, 22 -- References: {200705082228.l48MSFEO012827-at-ns.microscopy.com} 4, 22 -- In-Reply-To: {200705082228.l48MSFEO012827-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- X-UCalgary-MailScanner-Information: Please contact IT Help Desk at (403) 220-5555 for more information 4, 22 -- X-UCalgary-MailScanner: Found to be clean 4, 22 -- X-UCalgary-MailScanner-From: krensing-at-ucalgary.ca ==============================End of - Headers==============================
We have a Kevex Sigma Microanalyzer Level LPX3 which continually says that it is receiving no x-rays. The detector has recently been rebuilt and the system still does not work. Does anyone know of a company which repairs the Kevex electronics on site in Canada?
==============================Original Headers============================== 3, 21 -- From robinson32-at-sympatico.ca Tue May 8 19:20:59 2007 3, 21 -- Received: from bay0-omc3-s24.bay0.hotmail.com (bay0-omc3-s24.bay0.hotmail.com [65.54.246.224]) 3, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l490KxaF028288 3, 21 -- for {Microscopy-at-microscopy.com} ; Tue, 8 May 2007 19:20:59 -0500 3, 21 -- Received: from hotmail.com ([65.54.162.35]) by bay0-omc3-s24.bay0.hotmail.com with Microsoft SMTPSVC(6.0.3790.2668); 3, 21 -- Tue, 8 May 2007 17:20:58 -0700 3, 21 -- Received: from mail pickup service by hotmail.com with Microsoft SMTPSVC; 3, 21 -- Tue, 8 May 2007 17:20:58 -0700 3, 21 -- Message-ID: {BAY108-F250B140C816209D61AF92EBA3B0-at-phx.gbl} 3, 21 -- Received: from 65.54.162.200 by by108fd.bay108.hotmail.msn.com with HTTP; 3, 21 -- Wed, 09 May 2007 00:20:56 GMT 3, 21 -- X-Originating-IP: [70.54.69.188] 3, 21 -- X-Originating-Email: [robinson32-at-sympatico.ca] 3, 21 -- X-Sender: robinson32-at-sympatico.ca 3, 21 -- From: "L ROBINSON" {robinson32-at-sympatico.ca} 3, 21 -- To: Microscopy-at-microscopy.com 3, 21 -- Subject: EDS Repair 3, 21 -- Date: Wed, 09 May 2007 00:20:56 +0000 3, 21 -- Mime-Version: 1.0 3, 21 -- Content-Type: text/plain; format=flowed 3, 21 -- X-OriginalArrivalTime: 09 May 2007 00:20:58.0674 (UTC) FILETIME=[EACA6920:01C791CF] ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both rosslm-at-missouri.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: rosslm-at-missouri.edu Name: Lou Ross
Organization: University of Missouri-Columbia
Title-Subject: [Filtered] 5th Annual Short Course on Computer-Assisted Image Analysis
Question: The Electron Microscopy Core Facility at the University of Missouri-Columbia is hosting the 5th Annual Short Course and Workshop on Computer-Assisted Image Analysis and Measurement taught by Dr. John C. Russ on June 27-29, 2007. This popular course is intended to familiarize users of image analysis hardware and software with the fundamental principles and methods available to obtain meaningful results.
Image analysis and measurement techniques are utilized in a broad range of applications and are usually concerned with extracting a numerical values (number, size, shape, etc.) or location of objects from the image. In other cases, global structural parameters such as the volume and surface of features are of interest. These measurements may require image processing to correct defects, enhance features, compare multiple images, recognize objects, or other steps. Measurements on these individual features, or on the image as a whole, must then be obtained and interpreted in a proper stereological context to obtain useful data about the objects. Statistical interpretation of the data allows comparisons of different populations, understanding of distribution plots, and other inferences about the original objects. Structural modeling and geometric probability can be used to develop models for this interpretation.
The course relies heavily on tightly coupled lectures and hands-on exercises covering a wide variety of methodologies, approaches and tools, through a set of practical, step-by-step instructions to minimize the learning curve. No specific background is assumed, although users should already be familiar with microscopy or other imaging technologies. The software platform for the examples and exercises is Adobe Photoshop utilizing a comprehensive set of plug-ins from Reindeer Graphics. Included with the registration fee is a half-day primer on Photoshop prior to the beginning of the course.
Dr. Russ is the internationally acclaimed author of innumerable articles and several books on image analysis techniques and digital imaging, including the The Image Processing Handbook. He is widely known for his workshops and short courses and has helped to develop software packages to make image analysis methods more accessible to non-specialists. Many of the examples used in the course involve light or electron microscope images, but students are invited to bring their own images (TIFF files) for discussion and analysis. Ample time is allotted at the end of the course for individual instruction.
The registration fee is $1100. Enrollment is limited to 20 attendees and there are still a few openings. More information can be found at: http://www.emc.missouri.edu/works.htm or by contacting the course coordinator Lou Ross at 573.882.4777 or rosslm-at-missouri.edu.
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.com/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both frederic.diana-at-gmail.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: frederic.diana-at-gmail.com Name: Frederic
Organization: UCSB
Title-Subject: [Filtered] Autofocusing system
Question: Hi everyone!
I am looking for an autofocusing system which would be used for photoluminescence mapping. Basically I am looking for a system which can translate an objective lens to adjust the focus on the sample.
Depending of the size of the wafer, cristallographic oriention and direction you want to cut it, a solution could be to use a diamant wire saw, either to cut the sample or, better, to cut only two groves, where the silicon will (more or less cleanly, (100) works easier than (111)) clive. On such machines,it's very simple to cut parallel groves. Of coarse, if you have a 8" wafer, it will be less easy to find a saw which is big enought !
An other solution, if you have access to a power laser light, from the kind used for PLD (pulsed laser deposition), is to draw such groves with the laser. By that way you don't have any mechanical stress on the sample, and it will clive or brake very easy along the groves. It's very practical to have a wafer drawn with a grid patern, where one can brake squares or rectangles samples on demand. Of coarse, cristallography has its laws, and it brakes not always where one want !
Of coarse, in both cases, one makes the groves on the back side of the sample.
Hope it helps
-- J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
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Microscopy Folks,
I'd like to get some input on sample preparation with respect to silicon.
PROBLEM:
I have an FEI XL-50 FESEM that was originally designed to accept flat samples, essentially silicon wafers. It does not have what one would consider a typical exchange port. The exchange port is robotically controlled, and the maximum sample height, including stub, must be 5 mm or less. I must view these samples on edge, i.e. 90 degrees. This presents a problem in that I must cleave two parallel sections very close to each other. Perhaps diamond cutters can do this, but my hands are too shaky.
QUESTION:
Is there a device, or method, that would allow me to make these cleaves ~ 8 mm apart with any reasonable control? I found stubs that are low profile, 38 and 90 degrees, from the nice people at Ted Pella, but I still have this issue of doing two parallel cleaves very close together.
Just for info. purposes I am in the silicon MEMS development arena.
If you feel your reply is of general interest to this community, please reply to all, or you may contact me directly.
Regards to all in this small world,
Peter Tomic Renaissance Wireless Corp.
-- This email message is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies of the original message. If you are the intended recipient, please be advised that the content of this message is subject to access, review and disclosure by the sender's Email System Administrator.
==============================Original Headers============================== 13, 25 -- From peter.tomic-at-renwireless.com Tue May 8 12:33:06 2007 13, 25 -- Received: from mail.rw.local (4.piok2.xdsl.nauticom.net [209.195.176.101]) 13, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l48HX6om005343 13, 25 -- for {Microscopy-at-microscopy.com} ; Tue, 8 May 2007 12:33:06 -0500 13, 25 -- Received: from localhost (localhost [127.0.0.1]) 13, 25 -- by mail.rw.local (Postfix) with ESMTP id 97112241A0 13, 25 -- for {Microscopy-at-microscopy.com} ; Tue, 8 May 2007 13:33:05 -0400 (EDT) 13, 25 -- Received: from mail.rw.local ([127.0.0.1]) 13, 25 -- by localhost (mail.rw.local [127.0.0.1]) (amavisd-new, port 10024) with ESMTP 13, 25 -- id 30832-08 for {Microscopy-at-microscopy.com} ; 13, 25 -- Tue, 8 May 2007 13:33:05 -0400 (EDT) 13, 25 -- Received: from [127.0.0.1] (unknown [10.11.10.142]) 13, 25 -- by mail.rw.local (Postfix) with ESMTP id 2E11024168 13, 25 -- for {Microscopy-at-microscopy.com} ; Tue, 8 May 2007 13:33:05 -0400 (EDT) 13, 25 -- Message-ID: {4640B45E.6040307-at-renwireless.com} 13, 25 -- Date: Tue, 08 May 2007 13:33:18 -0400 13, 25 -- From: Peter Tomic {peter.tomic-at-renwireless.com} 13, 25 -- Organization: Renaissance Wireless Corp. 13, 25 -- User-Agent: Thunderbird 1.5.0.10 (Windows/20070221) 13, 25 -- MIME-Version: 1.0 13, 25 -- To: Microscopy-at-microscopy.com 13, 25 -- Subject: Silicon Cross-section sample preparation 13, 25 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 13, 25 -- Content-Transfer-Encoding: 7bit 13, 25 -- X-Virus-Scanned: amavisd-new at renwireless.com ==============================End of - Headers==============================
Hi Peter, I know you were working on III-V's a few years back so cleaving Si seems a lot more difficult in comparison. I have found the best way to cleave silicon wafers is rather different to GaAs or InP. Perhaps this is general knowledge and you know this already, but I hope it's worth sharing with the list if nothing else. The problem is that Si prefers to cleave along (111) rather than (110) and so you get an angled face on the cleave, which is usually rather uneven and often doesn't run straight. This is even worse when you are cleaving close to an existing edge, which attracts the crack front as it propagates (good for making low-angle cleaved specimens, but a problem for what you are trying to do). It is possible to make Si cleave along (110) by cleaving the wafer without any support. I suspect this works because the crack is propagating supersonically or something like that; I read a description of the technique in a paper from the 1970's but I can't remember the reference. So, to cleave Si along (110): make a single scribe mark on the top surface with a good sharp diamond a couple of mm long at the edge of the wafer. Then, hold the wafer just between forefinger and thumb in both hands, with the top wafer surface under your fingers and the scribe mark between the tips of your fingers. Put a thumbnail under the scribe mark and then bend the wafer down, pulling apart slightly at the same time. If it cleaves well, it will do so very quickly with an audible 'ping'. It's a good idea to do this over a large clean surface in case you drop either part. This is not so hard to do on a whole wafer (although trying not to drop an 8" wafer is fun, I haven't tried 300mm), but as the pieces get smaller it gets more difficult. It should be possible to get an 8mm wide strip, but a 5mm strip might be difficult or impossible. You could back thin the wafer but of course this isn't straightforward for something } 1" in diameter, and any scratches on the back might make the cleave deviate from its path. Like a lot of these things it's a lot easier to demonstrate than describe in text, and it takes a little practice to get the hang of it, so I would try on a few spare wafers first. I guess you could cleave a 10mm wide strip and grind it down to a couple of mm before mounting it for SEM.
-----Original Message----- X-from: peter.tomic-at-renwireless.com [mailto:peter.tomic-at-renwireless.com] Sent: 08 May 2007 18:35 To: Richard Beanland
Microscopy Folks,
I'd like to get some input on sample preparation with respect to silicon.
PROBLEM:
I have an FEI XL-50 FESEM that was originally designed to accept flat samples, essentially silicon wafers. It does not have what one would consider a typical exchange port. The exchange port is robotically controlled, and the maximum sample height, including stub, must be 5 mm or less. I must view these samples on edge, i.e. 90 degrees. This presents a problem in that I must cleave two parallel sections very close to each other. Perhaps diamond cutters can do this, but my hands are too shaky.
QUESTION:
Is there a device, or method, that would allow me to make these cleaves ~ 8 mm apart with any reasonable control? I found stubs that are low profile, 38 and 90 degrees, from the nice people at Ted Pella, but I still have this issue of doing two parallel cleaves very close together.
Just for info. purposes I am in the silicon MEMS development arena.
If you feel your reply is of general interest to this community, please reply to all, or you may contact me directly.
Regards to all in this small world,
Peter Tomic Renaissance Wireless Corp.
-- This email message is for the sole use of the intended recipient(s) and may contain confidential and privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies of the original message. If you are the intended recipient, please be advised that the content of this message is subject to access, review and disclosure by the sender's Email System Administrator.
==============================Original Headers============================== 13, 25 -- From peter.tomic-at-renwireless.com Tue May 8 12:33:06 2007 13, 25 -- Received: from mail.rw.local (4.piok2.xdsl.nauticom.net [209.195.176.101]) 13, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l48HX6om005343 13, 25 -- for {Microscopy-at-microscopy.com} ; Tue, 8 May 2007 12:33:06 -0500 13, 25 -- Received: from localhost (localhost [127.0.0.1]) 13, 25 -- by mail.rw.local (Postfix) with ESMTP id 97112241A0 13, 25 -- for {Microscopy-at-microscopy.com} ; Tue, 8 May 2007 13:33:05 -0400 (EDT) 13, 25 -- Received: from mail.rw.local ([127.0.0.1]) 13, 25 -- by localhost (mail.rw.local [127.0.0.1]) (amavisd-new, port 10024) with ESMTP 13, 25 -- id 30832-08 for {Microscopy-at-microscopy.com} ; 13, 25 -- Tue, 8 May 2007 13:33:05 -0400 (EDT) 13, 25 -- Received: from [127.0.0.1] (unknown [10.11.10.142]) 13, 25 -- by mail.rw.local (Postfix) with ESMTP id 2E11024168 13, 25 -- for {Microscopy-at-microscopy.com} ; Tue, 8 May 2007 13:33:05 -0400 (EDT) 13, 25 -- Message-ID: {4640B45E.6040307-at-renwireless.com} 13, 25 -- Date: Tue, 08 May 2007 13:33:18 -0400 13, 25 -- From: Peter Tomic {peter.tomic-at-renwireless.com} 13, 25 -- Organization: Renaissance Wireless Corp. 13, 25 -- User-Agent: Thunderbird 1.5.0.10 (Windows/20070221) 13, 25 -- MIME-Version: 1.0 13, 25 -- To: Microscopy-at-microscopy.com 13, 25 -- Subject: Silicon Cross-section sample preparation 13, 25 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 13, 25 -- Content-Transfer-Encoding: 7bit 13, 25 -- X-Virus-Scanned: amavisd-new at renwireless.com ==============================End of - Headers==============================
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==============================Original Headers============================== 23, 34 -- From richard.beanland-at-bookham.com Wed May 9 04:25:50 2007 23, 34 -- Received: from mail78.messagelabs.com (mail78.messagelabs.com [195.245.230.131]) 23, 34 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l499PnnN017010 23, 34 -- for {microscopy-at-microscopy.com} ; Wed, 9 May 2007 04:25:49 -0500 23, 34 -- X-VirusChecked: Checked 23, 34 -- X-Env-Sender: richard.beanland-at-bookham.com 23, 34 -- X-Msg-Ref: server-6.tower-78.messagelabs.com!1178702738!42757334!19 23, 34 -- X-StarScan-Version: 5.5.10.7.1; banners=bookham.com,-,- 23, 34 -- X-Originating-IP: [213.249.209.179] 23, 34 -- Received: (qmail 7787 invoked from network); 9 May 2007 09:25:48 -0000 23, 34 -- Received: from unknown (HELO cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI) (213.249.209.179) 23, 34 -- by server-6.tower-78.messagelabs.com with SMTP; 9 May 2007 09:25:48 -0000 23, 34 -- Received: from cas-smx-02.BOOKHAM.ENTERPRISE.PRI ([10.4.0.223]) by cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI with Microsoft SMTPSVC(6.0.3790.1830); 23, 34 -- Wed, 9 May 2007 10:26:47 +0100 23, 34 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 23, 34 -- Content-class: urn:content-classes:message 23, 34 -- MIME-Version: 1.0 23, 34 -- Content-Type: text/plain; 23, 34 -- charset="us-ascii" 23, 34 -- Subject: RE: [Microscopy] Silicon Cross-section sample preparation 23, 34 -- Date: Wed, 9 May 2007 10:26:46 +0100 23, 34 -- Message-ID: {9645D3E33E4C6548B12A7B25F611533E46C94C-at-cas-smx-02.BOOKHAM.ENTERPRISE.PRI} 23, 34 -- In-Reply-To: {200705081734.l48HYU4u007250-at-ns.microscopy.com} 23, 34 -- X-MS-Has-Attach: 23, 34 -- X-MS-TNEF-Correlator: 23, 34 -- Thread-Topic: [Microscopy] Silicon Cross-section sample preparation 23, 34 -- Thread-Index: AceRl5Y3SQeKavPPTXWrFceDxp3GzQAfFlIg 23, 34 -- References: {200705081734.l48HYU4u007250-at-ns.microscopy.com} 23, 34 -- From: "Richard Beanland" {richard.beanland-at-bookham.com} 23, 34 -- To: {peter.tomic-at-renwireless.com} 23, 34 -- Cc: {microscopy-at-microscopy.com} 23, 34 -- X-OriginalArrivalTime: 09 May 2007 09:26:47.0225 (UTC) FILETIME=[2A72BA90:01C7921C] 23, 34 -- Content-Transfer-Encoding: 8bit 23, 34 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l499PnnN017010 ==============================End of - Headers==============================
Yes, III-V compounds cleave so much easier, but now I'm working on Si MEMS. There's virtually no heat dissipation, therefore substrate thickness isn't an issue.
I've gotten several good suggestions, including yours. For the list, they are;
1. Laser serration
2. Thinning the Si [My samples are usually .025" thick.]
3. Wire Saw
4. Pre-scribing before cleaving
One other caveat in all this sample preparation is that the device constructed on top of the substrate is amorphous.
When all else fails, use an FIB, I always say.
I never get the easy problems.
Thanks to all of you.
Peter
Richard Beanland wrote: } Hi Peter, } I know you were working on III-V's a few years back so cleaving } Si seems a lot more difficult in comparison. I have found the best way } to cleave silicon wafers is rather different to GaAs or InP. Perhaps } this is general knowledge and you know this already, but I hope it's } worth sharing with the list if nothing else. } The problem is that Si prefers to cleave along (111) rather than (110) } and so you get an angled face on the cleave, which is usually rather } uneven and often doesn't run straight. This is even worse when you are } cleaving close to an existing edge, which attracts the crack front as it } propagates (good for making low-angle cleaved specimens, but a problem } for what you are trying to do). } It is possible to make Si cleave along (110) by cleaving the wafer } without any support. I suspect this works because the crack is } propagating supersonically or something like that; I read a description } of the technique in a paper from the 1970's but I can't remember the } reference. } So, to cleave Si along (110): make a single scribe mark on the top } surface with a good sharp diamond a couple of mm long at the edge of the } wafer. Then, hold the wafer just between forefinger and thumb in both } hands, with the top wafer surface under your fingers and the scribe mark } between the tips of your fingers. Put a thumbnail under the scribe mark } and then bend the wafer down, pulling apart slightly at the same time. } If it cleaves well, it will do so very quickly with an audible 'ping'. } It's a good idea to do this over a large clean surface in case you drop } either part. } This is not so hard to do on a whole wafer (although trying not to } drop an 8" wafer is fun, I haven't tried 300mm), but as the pieces get } smaller it gets more difficult. It should be possible to get an 8mm } wide strip, but a 5mm strip might be difficult or impossible. You could } back thin the wafer but of course this isn't straightforward for } something } 1" in diameter, and any scratches on the back might make the } cleave deviate from its path. Like a lot of these things it's a lot } easier to demonstrate than describe in text, and it takes a little } practice to get the hang of it, so I would try on a few spare wafers } first. I guess you could cleave a 10mm wide strip and grind it down to } a couple of mm before mounting it for SEM. } } Good luck! } } Richard } } ________________________________________ } Richard Beanland } Materials Analysis } Bookham } Caswell } Towcester } Northants } NN12 8EQ } United Kingdom } Tel. +44 1327 356362 } Fax. +44 1327 356775 } http://www.bookham.com } ________________________________________ } } -----Original Message----- } From: peter.tomic-at-renwireless.com [mailto:peter.tomic-at-renwireless.com] } Sent: 08 May 2007 18:35 } To: Richard Beanland } Subject: [Microscopy] Silicon Cross-section sample preparation } } } } } ------------------------------------------------------------------------ } ---- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ } ---- } } Microscopy Folks, } } I'd like to get some input on sample preparation with respect to } silicon. } } PROBLEM: } } I have an FEI XL-50 FESEM that was originally designed to accept flat } samples, essentially silicon wafers. It does not have what one would } consider a typical exchange port. The exchange port is robotically } controlled, and the maximum sample height, including stub, must be 5 mm } or less. I must view these samples on edge, i.e. 90 degrees. This } presents a problem in that I must cleave two parallel sections very } close to each other. Perhaps diamond cutters can do this, but my hands } are too shaky. } } QUESTION: } } Is there a device, or method, that would allow me to make these cleaves } ~ 8 mm apart with any reasonable control? I found stubs that are low } profile, 38 and 90 degrees, from the nice people at Ted Pella, but I } still have this issue of doing two parallel cleaves very close together. } } Just for info. purposes I am in the silicon MEMS development arena. } } If you feel your reply is of general interest to this community, please } reply to all, or you may contact me directly. } } Regards to all in this small world, } } Peter Tomic } Renaissance Wireless Corp. } }
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==============================Original Headers============================== 17, 27 -- From peter.tomic-at-renwireless.com Wed May 9 08:01:30 2007 17, 27 -- Received: from mail.rw.local (4.piok2.xdsl.nauticom.net [209.195.176.101]) 17, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l49D1T4U011473 17, 27 -- for {microscopy-at-microscopy.com} ; Wed, 9 May 2007 08:01:30 -0500 17, 27 -- Received: from localhost (localhost [127.0.0.1]) 17, 27 -- by mail.rw.local (Postfix) with ESMTP id 94A4D24251; 17, 27 -- Wed, 9 May 2007 09:01:29 -0400 (EDT) 17, 27 -- Received: from mail.rw.local ([127.0.0.1]) 17, 27 -- by localhost (mail.rw.local [127.0.0.1]) (amavisd-new, port 10024) with ESMTP 17, 27 -- id 17775-07; Wed, 9 May 2007 09:01:28 -0400 (EDT) 17, 27 -- Received: from [127.0.0.1] (unknown [10.11.10.142]) 17, 27 -- by mail.rw.local (Postfix) with ESMTP id 6DC2624240; 17, 27 -- Wed, 9 May 2007 09:01:28 -0400 (EDT) 17, 27 -- Message-ID: {4641C638.20301-at-renwireless.com} 17, 27 -- Date: Wed, 09 May 2007 09:01:44 -0400 17, 27 -- From: Peter Tomic {peter.tomic-at-renwireless.com} 17, 27 -- Organization: Renaissance Wireless Corp. 17, 27 -- User-Agent: Thunderbird 1.5.0.10 (Windows/20070221) 17, 27 -- MIME-Version: 1.0 17, 27 -- To: Richard Beanland {richard.beanland-at-bookham.com} 17, 27 -- CC: microscopy-at-microscopy.com 17, 27 -- Subject: Re: [Microscopy] Silicon Cross-section sample preparation 17, 27 -- References: {200705081734.l48HYU4u007250-at-ns.microscopy.com} {9645D3E33E4C6548B12A7B25F611533E46C94C-at-cas-smx-02.BOOKHAM.ENTERPRISE.PRI} 17, 27 -- In-Reply-To: {9645D3E33E4C6548B12A7B25F611533E46C94C-at-cas-smx-02.BOOKHAM.ENTERPRISE.PRI} 17, 27 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 17, 27 -- Content-Transfer-Encoding: 7bit 17, 27 -- X-Virus-Scanned: amavisd-new at renwireless.com ==============================End of - Headers==============================
June 24-27. 2007 University of California Santa Barbara
Register now and join other users of scanning probe microscopy as they meet and discuss their work informally with colleagues from all over the world at the fifth annual Seeing at the Nanoscale Conference, University of California, Santa Barbara. Sponsored by Veeco Instruments and the California NanoSystems Institute (CNSI), the conference includes two-and-one-half day technical presentations and a poster session on the following topics:
* Extending the Limits of SPM: High Speed Scanning, Ultra High Resolution Imaging, Multiple Probe SPM
* From Single Biomolecules To Cells: Using AFM and Combined AFM-Optical Techniques to Probe Biological Structures and Forces
* Next Generation Materials and Polymer Systems
* Beyond Topography: Measurement of Physical Properties at the Nanoscale - Nanomechanical, Electrical, Optical, Magnetic and Thermal
* Instruments and Probes - New Tools and Techniques for Nanoscience
For more information and to register, please go to www.veeco.com/Nanoconference
==============================Original Headers============================== 14, 20 -- From MCarlyle-at-veeco.com Wed May 9 10:57:19 2007 14, 20 -- Received: from sboexch2.int.veeco.com (SBOEXCH2.VEECO.com [68.111.35.167]) 14, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l49FvJZo026528 14, 20 -- for {Microscopy-at-Microscopy.com} ; Wed, 9 May 2007 10:57:19 -0500 14, 20 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 14, 20 -- content-class: urn:content-classes:message 14, 20 -- MIME-Version: 1.0 14, 20 -- Content-Type: text/plain; 14, 20 -- charset="us-ascii" 14, 20 -- Subject: SPM Users present cutting edge research 14, 20 -- Date: Wed, 9 May 2007 08:57:09 -0700 14, 20 -- Message-ID: {625BD6DD96DE554DBB0558FFE70E0F4201200565-at-sboexch2.int.veeco.com} 14, 20 -- X-MS-Has-Attach: 14, 20 -- X-MS-TNEF-Correlator: 14, 20 -- Thread-Topic: SPM Users present cutting edge research 14, 20 -- Thread-Index: AceSUrKJN3mIgLpjS+Oi37BgnFkPjw== 14, 20 -- From: "Marlene Carlyle" {MCarlyle-at-veeco.com} 14, 20 -- To: {Microscopy-at-Microscopy.com} 14, 20 -- Content-Transfer-Encoding: 8bit 14, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l49FvJZo026528 ==============================End of - Headers==============================
Depending on the size of the wafer, it will have either a flat or a notch on one place. This is the key to being parallel with the crystal lattice. So, align this key and use a carbide scribe (local hardware store or Home Depot--$5) and make two linear scribes then two lateral ones to get two small pieces of die. Mount on the Pella stubs and you should be good to go.
gary g.
At 09:35 AM 5/8/2007, you wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 6, 20 -- From gary-at-gaugler.com Wed May 9 11:34:24 2007 6, 20 -- Received: from qsmtp4.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 6, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l49GYOFq006116 6, 20 -- for {microscopy-at-microscopy.com} ; Wed, 9 May 2007 11:34:24 -0500 6, 20 -- Message-Id: {200705091634.l49GYOFq006116-at-ns.microscopy.com} 6, 20 -- Received: (qmail 1900 invoked from network); 9 May 2007 09:34:23 -0700 6, 20 -- Received: by simscan 1.1.0 ppid: 1894, pid: 1897, t: 0.1209s 6, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 6, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 6, 20 -- by qsmtp4 with SMTP; 9 May 2007 09:34:23 -0700 6, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.1.0.9 6, 20 -- Date: Wed, 09 May 2007 09:34:27 -0800 6, 20 -- To: peter.tomic-at-renwireless.com 6, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 6, 20 -- Subject: Re: [Microscopy] Silicon Cross-section sample preparation 6, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 6, 20 -- In-Reply-To: {200705081735.l48HZ5gi008227-at-ns.microscopy.com} 6, 20 -- References: {200705081735.l48HZ5gi008227-at-ns.microscopy.com} 6, 20 -- Mime-Version: 1.0 6, 20 -- Content-Type: text/plain; charset=us-ascii; format=flowed; x-avg-checked=avg-ok-C4C4242 ==============================End of - Headers==============================
During the last year I have attended sevaral demonstrations of whole slide image capture devices. For example, the TISSUEscope (Biomedical Photometrics) describes brightfield, fluorescence and confocal imaging and others - the Aperio - does brightfield imaging. There are others.
Does this emerging technology have a place in a core imaging facility - in other words, do users find that it replaces some other microscope modalities, e.g. I can see it very useful for measuring the frequency of rare events since they can see the entire slide.
My worry is that the data files are huge. Can analysis packages handle it easily? Do all computers have to be updated (64-bit, limitless RAM)? High throughput analysis to me implies speedy analysis, don't want to watch endless loading times, computer crashes.
I am curious about hearing any experience from users of whole slide scanners.
Thank you. Judy
Judy Trogadis Bio-Imaging Coordinator St. Michael's Hospital, 7Queen 30 Bond St. Toronto, ON M5B 1W8, Canada ph: 416-864-6060 x6337 pager: 416-685-9219 fax: 416-864-6043 trogadisj-at-smh.toronto.on.ca
==============================Original Headers============================== 9, 19 -- From trogadisj-at-smh.toronto.on.ca Wed May 9 11:49:31 2007 9, 19 -- Received: from smh.toronto.on.ca (mail.smh.toronto.on.ca [199.71.175.103]) 9, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l49GnUlX017680 9, 19 -- for {microscopy-at-microscopy.com} ; Wed, 9 May 2007 11:49:31 -0500 9, 19 -- Received: from ([172.27.15.58]) 9, 19 -- by beethoven.smh.toronto.on.ca with ESMTP id KP-GCT47.60776510; 9, 19 -- Wed, 09 May 2007 12:49:15 -0400 9, 19 -- Received: from SMTPOUT-MTA by beethoven.smh.toronto.on.ca 9, 19 -- with Novell_GroupWise; Wed, 09 May 2007 12:49:15 -0400 9, 19 -- Message-Id: {s641c34b.020-at-beethoven.smh.toronto.on.ca} 9, 19 -- X-Mailer: Novell GroupWise Internet Agent 6.5.7 9, 19 -- Date: Wed, 09 May 2007 12:48:50 -0400 9, 19 -- From: "Judy Trogadis" {TrogadisJ-at-smh.toronto.on.ca} 9, 19 -- To: {Microscopy-at-microscopy.com} 9, 19 -- Subject: microscopy-macroscopy 9, 19 -- Mime-Version: 1.0 9, 19 -- Content-Type: text/plain; charset=US-ASCII 9, 19 -- Content-Transfer-Encoding: 7bit 9, 19 -- Content-Disposition: inline ==============================End of - Headers==============================
I'm in desperate need of a copy of Sun Solaris OS 4.0.2 and a hard drive for a Sun SPARC IPC workstation. We have a Kratos XSAM 800 which is controlled by the SPARC computer, but unfortunately the computer has malfunctioned and we have no backup copies of the OS (the instrument was purchased "used"). I've already contacted Sun but they haven't been able to help. and I haven't received a reply from Kratos.
Can anyone on the list help?
Thanks in advance.
--Sue Kent
Susan M. Kent Principal Staff Scientist Continental AG Automotive Systems Division 21440 W. Lake Cook Road Deer Park, IL 60010 847-862-0216 (Desk) 847-343-5145 (Mobile) 847-862-8330 (Fax) Email: Susan.Kent-at-us.contiautomotive.com www.contiautomotive.com
______________________________________________________________________ This email has been scanned by the MessageLabs Email Security System. For more information please visit http://www.messagelabs.com/email ______________________________________________________________________
==============================Original Headers============================== 11, 32 -- From Susan.Kent-at-us.contiautomotive.com Wed May 9 14:11:43 2007 11, 32 -- Received: from mail153.messagelabs.com (mail153.messagelabs.com [216.82.253.51]) 11, 32 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l49JBhA9000463 11, 32 -- for {Microscopy-at-microscopy.com} ; Wed, 9 May 2007 14:11:43 -0500 11, 32 -- X-VirusChecked: Checked 11, 32 -- X-Env-Sender: Susan.Kent-at-us.contiautomotive.com 11, 32 -- X-Msg-Ref: server-5.tower-153.messagelabs.com!1178737902!8797097!1 11, 32 -- X-StarScan-Version: 5.5.10.7.1; banners=.,-,- 11, 32 -- X-Originating-IP: [129.188.136.8] 11, 32 -- Received: (qmail 12038 invoked from network); 9 May 2007 19:11:42 -0000 11, 32 -- Received: from motgate8.mot.com (HELO motgate8.mot.com) (129.188.136.8) 11, 32 -- by server-5.tower-153.messagelabs.com with SMTP; 9 May 2007 19:11:42 -0000 11, 32 -- Received: from il06exr01.mot.com (il06exr01.mot.com [129.188.137.131]) 11, 32 -- by motgate8.mot.com (8.12.11/Motorola) with ESMTP id l49JBg0x029085 11, 32 -- for {Microscopy-at-microscopy.com} ; Wed, 9 May 2007 12:11:42 -0700 (MST) 11, 32 -- Received: from il06vts03.mot.com (il06vts03.mot.com [129.188.137.143]) 11, 32 -- by il06exr01.mot.com (8.13.5/Vontu) with SMTP id l49JBfgJ025985 11, 32 -- for {Microscopy-at-microscopy.com} ; Wed, 9 May 2007 14:11:41 -0500 (CDT) 11, 32 -- Received: from dplc001.cig.mot.com (dplc001.cig.mot.com [10.21.43.133]) 11, 32 -- by il06exr01.mot.com (8.13.5/8.13.0) with ESMTP id l49JBf48025973 11, 32 -- for {Microscopy-at-microscopy.com} ; Wed, 9 May 2007 14:11:41 -0500 (CDT) 11, 32 -- Subject: XPS/ESCA: Need Help with Sun Hardware and Software 11, 32 -- To: Microscopy-at-microscopy.com 11, 32 -- X-Mailer: Lotus Notes Release 6.5.4 March 27, 2005 11, 32 -- Message-ID: {OF2B053001.63373870-ON862572D6.006830CD-862572D6.00696FCD-at-contiteves.com} 11, 32 -- From: Susan.Kent-at-us.contiautomotive.com 11, 32 -- Date: Wed, 9 May 2007 14:07:14 -0500 11, 32 -- X-MIMETrack: Serialize by Router on dplc001/srvc/na/au/cag(Release 6.5.6|March 06, 2007) at 11, 32 -- 05/09/2007 02:07:16 PM 11, 32 -- MIME-Version: 1.0 11, 32 -- Content-type: text/plain; charset=US-ASCII 11, 32 -- X-Vontu: Pass ==============================End of - Headers==============================
One of the major electron microscopy meetings this year will be held in the heart of the Inca empire. Machu Picchu, one of the world's most famous and wondrous places is nearby.
Leading microscopists from across the Americas (and from the rest of the world) will be presenting their latest work.
This is a reminder that abstracts (in a two-page format, very similar to the M and M format) are due by the end of this month (May, 2007).
Details of how to register (which must be done before the abstract is submitted) and of how to submit the abstract, can be found at the web site: http://www.ciasem2007.com/ (To change to English click the little blue button to the right.) The web site also lists invited speakers and gives details of the program. -- ........... 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
==============================Original Headers============================== 10, 21 -- From jae5-at-lehigh.edu Wed May 9 14:29:20 2007 10, 21 -- Received: from rain.CC.Lehigh.EDU (rain.CC.Lehigh.EDU [128.180.39.20]) 10, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l49JTJvu012168 10, 21 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 9 May 2007 14:29:20 -0500 10, 21 -- Received: from [127.0.0.1] (Dyn055141.mat.Lehigh.EDU [128.180.55.141]) 10, 21 -- (authenticated bits=0) 10, 21 -- by rain.CC.Lehigh.EDU (8.14.1/8.14.1) with ESMTP id l49JTI1x016376 10, 21 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NOT) 10, 21 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 9 May 2007 15:29:19 -0400 10, 21 -- Message-ID: {4642210E.6020408-at-lehigh.edu} 10, 21 -- Date: Wed, 09 May 2007 15:29:18 -0400 10, 21 -- From: Alwyn Eades {jae5-at-lehigh.edu} 10, 21 -- Organization: Lehigh University 10, 21 -- User-Agent: Thunderbird 1.5.0.10 (Windows/20070221) 10, 21 -- MIME-Version: 1.0 10, 21 -- To: MSA listserver {Microscopy-at-MSA.Microscopy.Com} 10, 21 -- Subject: 9th InterAmerican Congress on Electron Microscopy; Cusco, Peru 10, 21 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 10, 21 -- Content-Transfer-Encoding: 7bit 10, 21 -- X-Virus-Scanned: ClamAV 0.90.2/3224/Wed May 9 11:25:29 2007 on rain.CC.Lehigh.EDU 10, 21 -- X-Virus-Status: Clean ==============================End of - Headers==============================
I'm making subbed slides and just wondering if there is anyone who knows the chemistry/function of the chromium potassium sulfate (chrome alum) in the mix, and if there is a new-age environmentally-friendly substitute for it, or can it be eliminated - to what effect? I'm working on the assumption that it has a function. I know that only small quantities of Chrome alum are used, but would like to eliminate it if possible.
To avoid too much traffic about alternatives, let me say I've tried a number of other things for getting epoxy semi-thins to stick to glass, and this method works for me. I did already see a note in the archives about a "pinch of gelatine in the water bath" and that doesn't have the chromium compound. I've also read an alternative use of the Mayer's egg albumin - instead of smearing the slide and drying, adding some to the flotation water - and that also doesn't contain chromium. I like the way water drops sit nicely on the subbed slide so rows of sequential sections can be arrayed in the droplets without mixing.
Thanks in advance for any insights.
Dale Callaham
==============================Original Headers============================== 7, 19 -- From dac-at-research.umass.edu Wed May 9 15:29:53 2007 7, 19 -- Received: from race2.oit.umass.edu (race2.oit.umass.edu [128.119.101.38]) 7, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l49KTr08024803 7, 19 -- for {Microscopy-at-microscopy.com} ; Wed, 9 May 2007 15:29:53 -0500 7, 19 -- Received: from [172.30.55.164] (eutopia.bio.umass.edu [128.119.55.30]) 7, 19 -- (authenticated bits=0) 7, 19 -- by race2.oit.umass.edu (8.13.7/8.13.7) with ESMTP id l49KTq5g023611 7, 19 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NOT) 7, 19 -- for {Microscopy-at-microscopy.com} ; Wed, 9 May 2007 16:29:52 -0400 7, 19 -- Message-ID: {46423DAD.8060408-at-research.umass.edu} 7, 19 -- Date: Wed, 09 May 2007 16:31:25 -0500 7, 19 -- From: Dale Callaham {dac-at-research.umass.edu} 7, 19 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.0; en-US; rv:1.8.1.2) Gecko/20070222 SeaMonkey/1.1.1 7, 19 -- MIME-Version: 1.0 7, 19 -- To: Microscopy Listserver {Microscopy-at-microscopy.com} 7, 19 -- Subject: Gelatin subbed slides - eliminate the chromium? 7, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 7, 19 -- Content-Transfer-Encoding: 7bit 7, 19 -- X-Whitelist: TRUE ==============================End of - Headers==============================
I need to tap in to the wisdom of the list about dual beam argon ion mills.
We have a Bal-Tec RES100 ion mill that we are having difficulty maintaining. It had been in disuse for a couple of years, and has been repaired and upgraded recently, at the factory, to be as close to a RES101 as possible. We are having issues such as the guns becoming contaminated and needing service much more frequently than we think they should. Routine maintenance seems to be difficult, too. We think we are looking at a high maintenance instrument here. I would like to know what kind of experience other labs have had with this instrument.
I would also like to hear from people who have experience using/maintaining the RES100/101 or the Gatan PIPS, or both. We need some comparative information, so we can make a decision about how to proceed with our ion milling needs.
Any input would be appreciated.
--John
John Chandler Manager, EM Lab Colorado School of Mines Golden, CO 80401 jpchandl-at-mines.edu 303-384-2203
==============================Original Headers============================== 9, 19 -- From jpchandl-at-mines.edu Wed May 9 15:42:15 2007 9, 19 -- Received: from inspire.Mines.EDU (inspire.Mines.EDU [138.67.130.5]) 9, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l49KgEDO003975 9, 19 -- for {microscopy-at-microscopy.com} ; Wed, 9 May 2007 15:42:14 -0500 9, 19 -- Received: from emlab1 (emlab1.Mines.EDU [138.67.28.173]) 9, 19 -- by inspire.Mines.EDU (8.13.1/8.13.1) with ESMTP id l49KgD7T016786 9, 19 -- for {microscopy-at-microscopy.com} ; Wed, 9 May 2007 14:42:13 -0600 9, 19 -- From: "John Chandler" {jpchandl-at-mines.edu} 9, 19 -- To: {microscopy-at-microscopy.com} 9, 19 -- Subject: Looking for recommendations about dual beam ion mills 9, 19 -- Date: Wed, 9 May 2007 14:42:13 -0600 9, 19 -- Message-ID: {001e01c7927a$861c8570$ad1c438a-at-mines.edu} 9, 19 -- MIME-Version: 1.0 9, 19 -- Content-Type: text/plain; 9, 19 -- charset="us-ascii" 9, 19 -- Content-Transfer-Encoding: 7bit 9, 19 -- X-Mailer: Microsoft Office Outlook 11 9, 19 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 9, 19 -- Thread-Index: AceSeoX7iyPEBSG3T3O4yAVZe0waUw== ==============================End of - Headers==============================
If you haven't tried coating your slides with 1% aminopropyltriethoxysilane (APTS) in water then you are missing out on a much better alternative. The water really beads up on it. I used to use chrome-gel but it doesn't compare.
At 03:30 PM 05/09/07, you wrote:
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
Dale- Have you tried the SuperFrost Plus slides? They are pre-treated with something proprietary that helps sections adhere. They can be purchased from any number of vendors, although I believe that they all come from the same manufacturer. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Weill-Cornell Medical College
} } } Hi all, } } } } } } I am trying to find resolution of a problem with our CM300 TEM with a } } } LaB6 cathode. } } } } } } When trying to get diffraction from an almost amorphous material I } } } have noticed a presence of relatively weak but clearly visible } } } perfectly circular ring of intensity around the central transmitted } } } spot. The ring is not centered around the transmitted spot. Its } } } position varies when operating the beam shift controls, its size } } } varies with the C2 aperture size when in diffraction mode. It is also } } } present when there is no sample under the beam. When there is } } } strongly reflecting crystalline material the ring is almost invisible } } } due to its weak relative intensity. It is present at any accelerating } } } voltage. } } } } } } I am trying to get some ideas about what might be causing the ring } } } and how to eliminate it. } } } } } } The ring is also visible when in LM imaging mode (image is formed by } } } the diffraction lens) } } } and when the beam is focused to a spot. In LM mode the ring has } } } strange shape. It has sharp circular outline on its outer edge and it } } } has irregular shape and outline on its inner edge. } } } } } } Using the free lens control option I have made the following } } } observations: } } } } } } In LM imaging mode with beam focused to a spot. } } } When changing the C1 lens current -the ring changes size and focus } } } but does not rotate. } } } changing C2 lens current - change in focus only, plus some rotation } } } changing Twin lens current - change of size and focus, no rotation } } } changing Objective lens current - change in size and focus, some } } } rotation } } } changing Diffraction lens current - change in size and focus, no } } } rotation } } } changing Intermediate lens current - change in size and focus, no } } } rotation } } } changing P1 lens current - change in size and focus, some rotation } } } changing P2 lens current - rotation only } } } } } } In diffraction mode with fully spread beam: } } } position varies when operating the beam shift controls } } } size varies with the C2 aperture size. } } } } } } FEI support engineers have not been succesful in identifing the } } } problem so far. It was suggested that it is due to undersaturated } } } LaB6 cathode, a possibility which we have clearly eliminated as a } } } possible source. } } } } } } Thanks for any hints our suggestions. } } } } } } Krassimir. } } } _______________________________________ } } } Krassimir N. Bozhilov } } } Central Facility for Advanced Microscopy and Microanalysis } } } University of California } } } Riverside, CA 92521 } } } } } } tel 951 827 2998 } } } fax 951 827 2489 } } } bozhilov-at-ucr.edu } } } _______________________________________ } } } } On May 8, 2007, at 12:46 PM, randerson20-at-tampabay.rr.com wrote: } } } } } } ---------------------------------------------------------------------- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } } America } } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } } MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------- } } } } Consider the possibility that it might be visible light from the hot } } cathode coming down the column, i.e. like a flashlight. } } } } This argument is abetted by the sharp outside edge, from apertures } } along } } the way, and a diffuse inner edge and the fact that you see it } } without a } } specimen loaded. However, it isn't immediately obvious how or why it } } changes size and shape with all of your other perturbations. } } } } Can you partly eclipse it by moving apertures around? } } } } Ron Anderson } } } } bozhilov-at-ucr.edu wrote: } } } --------------------------------------------------------------------- } } } ------- } } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } } } America } } } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } } } MicroscopyListserver } } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } --------------------------------------------------------------------- } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
The fact that the ring changes as the lenses are adjusted suggests that it is not anything to do with light from the filament.
Since the ring changes in dia. as the C2 aperture is changed suggests that it is something to do with the C2 aperture.
Do you have an unusually high X-ray background?
Do you clean your C2 apertures yourself?
I'm thinking that the C2 apertures have been overheated in the cleaning process, leading to recrystalisation and a rough edge to the inside bore of the apertures?
You might be able to confirm this by doing convergent beam diffraction - the ragged edge of the C2 aperture should be visible in the CBDP.
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==============================Original Headers============================== 9, 16 -- From larry-at-celtic.freewire.co.uk Wed May 9 16:05:24 2007 9, 16 -- Received: from get.freewire.net (get.freewire.net [86.54.106.202] (may be forged)) 9, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l49L5O5q006033 9, 16 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 9 May 2007 16:05:24 -0500 9, 16 -- Received: from [217.154.251.60] (th6dc-217-154-251-60.dial.mistral.co.uk [217.154.251.60] (may be forged)) 9, 16 -- by get.freewire.net (8.13.6/8.11.6) with ESMTP id l49L5IHF025740 9, 16 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 9 May 2007 22:05:20 +0100 9, 16 -- Mime-Version: 1.0 9, 16 -- Message-Id: {p06210200c267e4f0f513-at-[217.154.253.101]} 9, 16 -- In-Reply-To: {200705082125.l48LP5XL021706-at-ns.microscopy.com} 9, 16 -- References: {200705082125.l48LP5XL021706-at-ns.microscopy.com} 9, 16 -- Date: Wed, 9 May 2007 21:59:29 +0100 9, 16 -- To: Microscopy-at-MSA.Microscopy.Com 9, 16 -- From: Larry Stoter {larry-at-celtic.freewire.co.uk} 9, 16 -- Subject: Re: [Microscopy] Diffracation ring problem 9, 16 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
You do not need the chrome alum, but the slides will be "stickier" with it. I have no idea why....
I'd agree with the rest of the crowd about APS or Plus slides, but sometimes they just aren't enough & subbing is still the way to go, horrible as the process may be.
Tamara
On Wed, 9 May 2007 15:31:13 -0500 dac-at-research.umass.edu wrote: } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy } Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Hi everyone, } } I'm making subbed slides and just wondering if there is } anyone who knows } the chemistry/function of the chromium potassium sulfate } (chrome alum) } in the mix, and if there is a new-age } environmentally-friendly } substitute for it, or can it be eliminated - to what } effect? I'm working } on the assumption that it has a function. I know that } only small } quantities of Chrome alum are used, but would like to } eliminate it if } possible. } } To avoid too much traffic about alternatives, let me say } I've tried a } number of other things for getting epoxy semi-thins to } stick to glass, } and this method works for me. I did already see a note } in the archives } about a "pinch of gelatine in the water bath" and that } doesn't have the } chromium compound. I've also read an alternative use of } the Mayer's egg } albumin - instead of smearing the slide and drying, } adding some to the } flotation water - and that also doesn't contain } chromium. I like the way } water drops sit nicely on the subbed slide so rows of } sequential } sections can be arrayed in the droplets without mixing. } } } Thanks in advance for any insights. } } } Dale Callaham } } ==============================Original } Headers============================== } 7, 19 -- From dac-at-research.umass.edu Wed May 9 15:29:53 } 2007 } 7, 19 -- Received: from race2.oit.umass.edu } (race2.oit.umass.edu [128.119.101.38]) } 7, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) } with ESMTP id l49KTr08024803 } 7, 19 -- for {Microscopy-at-microscopy.com} ; Wed, 9 May } 2007 15:29:53 -0500 } 7, 19 -- Received: from [172.30.55.164] } (eutopia.bio.umass.edu [128.119.55.30]) } 7, 19 -- (authenticated bits=0) } 7, 19 -- by race2.oit.umass.edu (8.13.7/8.13.7) with } ESMTP id l49KTq5g023611 } 7, 19 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA } bits=256 verify=NOT) } 7, 19 -- for {Microscopy-at-microscopy.com} ; Wed, 9 May } 2007 16:29:52 -0400 } 7, 19 -- Message-ID: } {46423DAD.8060408-at-research.umass.edu} } 7, 19 -- Date: Wed, 09 May 2007 16:31:25 -0500 } 7, 19 -- From: Dale Callaham {dac-at-research.umass.edu} } 7, 19 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT } 5.0; en-US; rv:1.8.1.2) Gecko/20070222 SeaMonkey/1.1.1 } 7, 19 -- MIME-Version: 1.0 } 7, 19 -- To: Microscopy Listserver } {Microscopy-at-microscopy.com} } 7, 19 -- Subject: Gelatin subbed slides - eliminate the } chromium? } 7, 19 -- Content-Type: text/plain; charset=ISO-8859-1; } format=flowed } 7, 19 -- Content-Transfer-Encoding: 7bit } 7, 19 -- X-Whitelist: TRUE } ==============================End of - } Headers==============================
I agree with the other comments: Ferrocyanide, decreasing dehydration times (penetration is immediate in monolayers) and using picric acid probably can improve membrane contrast. (personally I already tried UAc in methanol without satisfaction)
Please share your experience with us.
Best regards,
Stephane
--- tbargar-at-unmc.edu wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } } Dear listers, } } Anyone out there have any advice on how to enhance } the contrast and } definition of the membranes of the cristae of } mitochondria? The samples } brought to me are monolayer cell cultures of cancer } cells grown on } Thermanox coverslips. This is how I'm currently } processing the samples: } Primary fixation is 2% glutaraldehyde, 2% } paraformaldehyde, 0.5% acrolein } in 0.1M Sorensen's phosphate buffer pH 7.2. } Post-fixation in 1%Osmium } Tetroxide in 0.1M Sorensen's phosphate buffer. } Dehydration in 50, 70, 90, } 95, 100%X3 ethanol solutions. embedding in } Araldite. Sections are stained } with 2% uranyl acetate aqueous 15 minutes and } Reynold's lead citrate 10 } minutes. The density of the cytoplasm and } mitochondrial matrix are } similar with the result that the contrast of the } mitochondria is similar to } the cytoplasm. The mitochondria and it's membranes } (outer and that of the } cristae) don't really "stand out". The researchers } involved want to see } really contrasty mitochondrial cristae. } } The next thing I'm going to try is post-fixation } with a mix of osmium } tetroxide and potassium ferrocyanide. } } Anyone have any other ideas? Thanks in advance. } } } Tom Bargar } University of Nebraska Medical Center } Core Electron Microscopy Research Facility } 986395 Nebraska Medical Center } Omaha, NE 68198-6395 } 402-559-7347 } tbargar-at-unmc.edu } } } ==============================Original } Headers============================== } 8, 20 -- From tbargar-at-unmc.edu Tue May 1 15:39:39 } 2007 } 8, 20 -- Received: from zixvpm02.unmc.edu } (zixvpm02.unmc.edu [192.198.54.127]) } 8, 20 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with ESMTP id } l41KddQH023357 } 8, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, } 1 May 2007 15:39:39 -0500 } 8, 20 -- Received: from zixvpm02.unmc.edu (ZixVPM } [127.0.0.1]) } 8, 20 -- by Outbound.unmc.edu (Proprietary) with } ESMTP id 8A304A0061 } 8, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, } 1 May 2007 15:39:39 -0500 (CDT) } 8, 20 -- Received: from unmcnotes01.unmc.edu } (host-137-197-103-35.unmc.edu [137.197.103.35]) } 8, 20 -- by zixvpm02.unmc.edu (Proprietary) with } ESMTP id 72E1039804A } 8, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, } 1 May 2007 15:39:38 -0500 (CDT) } 8, 20 -- Subject: Help with enhancing contrast of } Mitochondrial cristae } 8, 20 -- To: Microscopy-at-MSA.Microscopy.Com } 8, 20 -- X-Mailer: Lotus Notes Release 7.0.1 January } 17, 2006 } 8, 20 -- Message-ID: } {OFDC8670F1.FEB83264-ON862572CE.006F5771-862572CE.00717D6E-at-unmc.edu} } 8, 20 -- From: Tom W Bargar {tbargar-at-unmc.edu} } 8, 20 -- Date: Tue, 1 May 2007 15:39:27 -0500 } 8, 20 -- X-MIMETrack: Serialize by Router on } UNMCNOTES01.UNMC.EDU/Servers/UNEBR at 05/01/2007 } 03:39:38 } 8, 20 -- PM } 8, 20 -- MIME-Version: 1.0 } 8, 20 -- Content-type: text/plain; charset=US-ASCII } ==============================End of - } Headers============================== }
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==============================Original Headers============================== 9, 21 -- From nizets2-at-yahoo.com Thu May 10 06:15:21 2007 9, 21 -- Received: from web37402.mail.mud.yahoo.com (web37402.mail.mud.yahoo.com [209.191.91.134]) 9, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l4ABFKJw009245 9, 21 -- for {microscopy-at-microscopy.com} ; Thu, 10 May 2007 06:15:21 -0500 9, 21 -- Received: (qmail 27735 invoked by uid 60001); 10 May 2007 11:15:20 -0000 9, 21 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 9, 21 -- s=s1024; d=yahoo.com; 9, 21 -- h=X-YMail-OSG:Received:Date:From:Subject:To:Cc:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding:Message-ID; 9, 21 -- b=xdVFiLCF4Y+sO8r3CpubJCf2uWbS/tsa8x0ab/hPl+5HFUOENjJ6yIvxMiFISE4+rsVrEo+9g9amUFa4qIQOAGV0FBzymuaE0sNVDp0SglZljLy8sAz3+WIuaj+c/D2DZnTf4n+Vivh1RJ9XqSQvZy3HvK1FSpB028byJ8YI300=; 9, 21 -- X-YMail-OSG: mktHeDIVM1kftlmU8fPc3P3OX17YmyvCJT352ac98PdSr0qhC7X.fCqkPbxI6mkxa09GcWQMmEhQCWAHO8TzXOi4W.ueJ.8eBX54nP.Y7I3f9c1fcF6eyrST0RA3CoDbTEtqWbHpql.Ve33A65i7tH.ZVhYJ5Jn8TfeDlO7pDP4W.DeEznuXRaE- 9, 21 -- Received: from [80.122.101.102] by web37402.mail.mud.yahoo.com via HTTP; Thu, 10 May 2007 04:15:20 PDT 9, 21 -- Date: Thu, 10 May 2007 04:15:20 -0700 (PDT) 9, 21 -- From: Stephane Nizet {nizets2-at-yahoo.com} 9, 21 -- Subject: Re: [Microscopy] Help with enhancing contrast of Mitochondrial cristae 9, 21 -- To: tbargar-at-unmc.edu 9, 21 -- Cc: microscopy-at-microscopy.com 9, 21 -- In-Reply-To: {200705012045.l41Kjoma031136-at-ns.microscopy.com} 9, 21 -- MIME-Version: 1.0 9, 21 -- Content-Type: text/plain; charset=iso-8859-1 9, 21 -- Content-Transfer-Encoding: 8bit 9, 21 -- Message-ID: {635982.26761.qm-at-web37402.mail.mud.yahoo.com} ==============================End of - Headers==============================
I just thought I should mention something that I think I read years ago in one of M.A. Hayat's books. He mentions that cacodylate and some other buffers when incorporated into a fixative produce higher contrast because they are more extractive whereas phosphate buffers probably preserve cell contents better but at the expense of contrast.
I just thought that I should add this because you mention Sorenson's buffer in the fixative. I'm sure that many of the other points have an effect but just wondered if the buffer could be contributing to your problems as well.
Malcolm
Malcolm Haswell e.m. unit School of Health, Natural and Social Sciences Fleming Building University of Sunderland Tyne & Wear SR1 3SD UK e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- X-from: nizets2-at-yahoo.com
John
I work with four Gatan PIPS daily and have found them to be quite acceptable in ease of both maintenance and use.
We currently do maintenance only when the ion guns stop functioning, at that point the machine is taken out of service and a full cleaning performed. This happens about every five to six months and leaves the PIPS out of service for two days (One day to clean, pump down over night, and alignment the next day). The diaphragm pump needs new diaphragms once a year (about an hours work) but not too much else.
Work load is typically one to two ceramic samples per day, with an occasional silicon or glass sample thrown in. The ceramic samples are lapped to about six microns and mill for an hour to two hours. Silicon and glass are left about ten microns thick and take much longer (up to three days), not so much due to the thickness as to the low kVs used and letting the sample "cool".
Gatan offers a digital camera/zoom lens set and an external diaphragm pump as extras. In my opinion, they are a "must have".
If you have any further questions please feel free to contact me.
Neal R. Zimmermann
On Wed, 2007-05-09 at 15:43 -0500, jpchandl-at-mines.edu wrote: } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } I need to tap in to the wisdom of the list about dual beam argon ion mills. } } We have a Bal-Tec RES100 ion mill that we are having difficulty maintaining. } It had been in disuse for a couple of years, and has been repaired and } upgraded recently, at the factory, to be as close to a RES101 as possible. } We are having issues such as the guns becoming contaminated and needing } service much more frequently than we think they should. Routine maintenance } seems to be difficult, too. We think we are looking at a high maintenance } instrument here. I would like to know what kind of experience other labs } have had with this instrument. } } I would also like to hear from people who have experience using/maintaining } the RES100/101 or the Gatan PIPS, or both. We need some comparative } information, so we can make a decision about how to proceed with our ion } milling needs. } } Any input would be appreciated. } } --John } } John Chandler } Manager, EM Lab } Colorado School of Mines } Golden, CO 80401 } jpchandl-at-mines.edu } 303-384-2203 } } } } } ==============================Original Headers============================== } 9, 19 -- From jpchandl-at-mines.edu Wed May 9 15:42:15 2007 } 9, 19 -- Received: from inspire.Mines.EDU (inspire.Mines.EDU [138.67.130.5]) } 9, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l49KgEDO003975 } 9, 19 -- for {microscopy-at-microscopy.com} ; Wed, 9 May 2007 15:42:14 -0500 } 9, 19 -- Received: from emlab1 (emlab1.Mines.EDU [138.67.28.173]) } 9, 19 -- by inspire.Mines.EDU (8.13.1/8.13.1) with ESMTP id l49KgD7T016786 } 9, 19 -- for {microscopy-at-microscopy.com} ; Wed, 9 May 2007 14:42:13 -0600 } 9, 19 -- From: "John Chandler" {jpchandl-at-mines.edu} } 9, 19 -- To: {microscopy-at-microscopy.com} } 9, 19 -- Subject: Looking for recommendations about dual beam ion mills } 9, 19 -- Date: Wed, 9 May 2007 14:42:13 -0600 } 9, 19 -- Message-ID: {001e01c7927a$861c8570$ad1c438a-at-mines.edu} } 9, 19 -- MIME-Version: 1.0 } 9, 19 -- Content-Type: text/plain; } 9, 19 -- charset="us-ascii" } 9, 19 -- Content-Transfer-Encoding: 7bit } 9, 19 -- X-Mailer: Microsoft Office Outlook 11 } 9, 19 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 } 9, 19 -- Thread-Index: AceSeoX7iyPEBSG3T3O4yAVZe0waUw== } ==============================End of - Headers==============================
==============================Original Headers============================== 11, 24 -- From nealzimm-at-cpinternet.com Thu May 10 08:08:28 2007 11, 24 -- Received: from cpinternet.com (mail.cpinternet.com [209.240.224.81]) 11, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4AD8S9W001596 11, 24 -- for {microscopy-at-microscopy.com} ; Thu, 10 May 2007 08:08:28 -0500 11, 24 -- Received: from [192.168.1.102] (unverified [216.251.191.121]) 11, 24 -- by cpinternet.com (SurgeMail 3.8j) with ESMTP id 171217060-1881964 11, 24 -- for multiple; Thu, 10 May 2007 08:08:27 -0500 CDT 11, 24 -- Subject: Re: [Microscopy] Looking for recommendations about dual beam ion 11, 24 -- mills 11, 24 -- From: Neal Zimmermann {nealzimm-at-cpinternet.com} 11, 24 -- To: jpchandl-at-mines.edu, microscopy-at-microscopy.com 11, 24 -- In-Reply-To: {200705092043.l49Kh2W8005876-at-ns.microscopy.com} 11, 24 -- References: {200705092043.l49Kh2W8005876-at-ns.microscopy.com} 11, 24 -- Content-Type: text/plain 11, 24 -- Date: Thu, 10 May 2007 08:08:14 -0500 11, 24 -- Message-Id: {1178802494.3663.19.camel-at-localhost.localdomain} 11, 24 -- Mime-Version: 1.0 11, 24 -- X-Mailer: Evolution 2.8.3 (2.8.3-2.fc6) 11, 24 -- Content-Transfer-Encoding: 7bit 11, 24 -- X-Authenticated-User: nealzimm-at-cpinternet.com 11, 24 -- X-NotAscii: charset=us-ascii 11, 24 -- X-Encryption: SSL encrypted 11, 24 -- X-IP-stats: No info recorded yet ip=216.251.191.121 11, 24 -- X-Originating-IP: 216.251.191.121 ==============================End of - Headers==============================
I got my recipe for chrome-alum from an old copy of Berlin and Miksche's "Botanical Microtechnique and Cytochemistry". In the chapter covering microtomy, in the section under adhesives, there is a subsection for gelatin adhesives. The first recipe is for "Haupt's adhesive". This is prepared by dissolving 1g of gelatine in 100 cc water, smearing a thin film of this onto slides, and then flooding with 4% formalin. The second recipe is for the chorme-alum that we all know and love. The paragraph which introduces the chrome-alum adhesive technique begins: "A gelatin adhesive that does not use formalin may be prepared and used as follows...".
I know I'm telling this poorly, but the point is that I think the chrome-alum is used to "fix" the gelatine in place in a manner analogous to what the formaldehyde would do in the Haupt's solution. This is the purpose (I believe) in using chrome in the tanning industry. Furthermore, the treatment of the gelatine layer with formaldehyde or chrome would then leave free aldehydes/uncomplexed chrome atoms embedded in the gelatine which might then react with your sections, adhering them to the slide. Using glutaraldehyde instead of formaldehyde might even provide a stronger bond, if that is what you decide to go with.
So here is a possible reason for the chrome-alum and a possible (although old-school) alternative that still uses gelatine as the adhesive but does not contain chromium.
Andy Bowling Plant Physiologist USDA-ARS-SWSRU
==============================Original Headers============================== 5, 30 -- From Andrew.Bowling-at-ARS.USDA.GOV Thu May 10 08:38:50 2007 5, 30 -- Received: from messagescreen4.ars.usda.gov (messagescreen4.ars.usda.gov [199.133.180.151]) 5, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4ADcn3d013639 5, 30 -- for {microscopy-at-microscopy.com} ; Thu, 10 May 2007 08:38:49 -0500 5, 30 -- Received: from CO-MAILBH-02.ARSNET.ARS.USDA.GOV ([199.133.183.227]) 5, 30 -- by messagescreen4.ars.usda.gov (8.13.8/8.13.8) with ESMTP id l4ADcnMH021136 5, 30 -- for {microscopy-at-microscopy.com} ; Thu, 10 May 2007 08:38:49 -0500 5, 30 -- Received: from CO-MAIL-03.ARSNET.ARS.USDA.GOV ([10.100.2.202]) by CO-MAILBH-02.ARSNET.ARS.USDA.GOV with Microsoft SMTPSVC(6.0.3790.3959); 5, 30 -- Thu, 10 May 2007 07:38:49 -0600 5, 30 -- Content-class: urn:content-classes:message 5, 30 -- MIME-Version: 1.0 5, 30 -- Content-Type: text/plain; 5, 30 -- charset="us-ascii" 5, 30 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 5, 30 -- Subject: RE: Gelatin subbed slides - eliminate the chromium? 5, 30 -- Date: Thu, 10 May 2007 07:38:49 -0600 5, 30 -- Message-ID: {8017F94146BF634DA9414E4B9088525B07F382-at-CO-MAIL-03.ARSNET.ARS.USDA.GOV} 5, 30 -- X-MS-Has-Attach: 5, 30 -- X-MS-TNEF-Correlator: 5, 30 -- Thread-Topic: RE: Gelatin subbed slides - eliminate the chromium? 5, 30 -- Thread-Index: AceTEOhcx6ZpfXvXRquZ2KUGmRxDUg== 5, 30 -- From: "Bowling, Andrew" {Andrew.Bowling-at-ARS.USDA.GOV} 5, 30 -- To: {microscopy-at-microscopy.com} 5, 30 -- X-OriginalArrivalTime: 10 May 2007 13:38:49.0402 (UTC) FILETIME=[8A6191A0:01C79308] 5, 30 -- X-MessageScreenMessageID: 1178804329.932632.1254.3150888985 5, 30 -- X-MessageScreenContentScore: Score of 0 assigned to Content 5, 30 -- X-MessageScreenUCEScore: Score of 0 assigned to UCE 5, 30 -- X-MessageScreen: Analyzed by IntelliReach MessageScreen(tm) 5, 30 -- Content-Transfer-Encoding: 8bit 5, 30 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4ADcn3d013639 ==============================End of - Headers==============================
Neal, I wouldn't expect a 10um Si sample to take more than 30-60 mins to thin in a PIPS. I can only think you are working at a relatively high milling angle (} 5 degrees) and so have to turn the beam energy down very low so you don't damage your sample. Why not experiment with one sample using full power (6kV) and 3 degrees incidence angle, finishing off with 10 mins at 2kV? It could increase your throughput more than 20 times!
-----Original Message----- X-from: nealzimm-at-cpinternet.com [mailto:nealzimm-at-cpinternet.com] Sent: 10 May 2007 14:10 To: Richard Beanland
John
I work with four Gatan PIPS daily and have found them to be quite acceptable in ease of both maintenance and use.
We currently do maintenance only when the ion guns stop functioning, at that point the machine is taken out of service and a full cleaning performed. This happens about every five to six months and leaves the PIPS out of service for two days (One day to clean, pump down over night, and alignment the next day). The diaphragm pump needs new diaphragms once a year (about an hours work) but not too much else.
Work load is typically one to two ceramic samples per day, with an occasional silicon or glass sample thrown in. The ceramic samples are lapped to about six microns and mill for an hour to two hours. Silicon and glass are left about ten microns thick and take much longer (up to three days), not so much due to the thickness as to the low kVs used and letting the sample "cool".
Gatan offers a digital camera/zoom lens set and an external diaphragm pump as extras. In my opinion, they are a "must have".
If you have any further questions please feel free to contact me.
Neal R. Zimmermann
On Wed, 2007-05-09 at 15:43 -0500, jpchandl-at-mines.edu wrote: } } } ------------------------------------------------------------------------ ---- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ ---- } } I need to tap in to the wisdom of the list about dual beam argon ion mills. } } We have a Bal-Tec RES100 ion mill that we are having difficulty maintaining. } It had been in disuse for a couple of years, and has been repaired and } upgraded recently, at the factory, to be as close to a RES101 as possible. } We are having issues such as the guns becoming contaminated and needing } service much more frequently than we think they should. Routine maintenance } seems to be difficult, too. We think we are looking at a high maintenance } instrument here. I would like to know what kind of experience other labs } have had with this instrument. } } I would also like to hear from people who have experience using/maintaining } the RES100/101 or the Gatan PIPS, or both. We need some comparative } information, so we can make a decision about how to proceed with our ion } milling needs. } } Any input would be appreciated. } } --John } } John Chandler } Manager, EM Lab } Colorado School of Mines } Golden, CO 80401 } jpchandl-at-mines.edu } 303-384-2203 } } } } } ==============================Original Headers============================== } 9, 19 -- From jpchandl-at-mines.edu Wed May 9 15:42:15 2007 } 9, 19 -- Received: from inspire.Mines.EDU (inspire.Mines.EDU [138.67.130.5]) } 9, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l49KgEDO003975 } 9, 19 -- for {microscopy-at-microscopy.com} ; Wed, 9 May 2007 15:42:14 -0500 } 9, 19 -- Received: from emlab1 (emlab1.Mines.EDU [138.67.28.173]) } 9, 19 -- by inspire.Mines.EDU (8.13.1/8.13.1) with ESMTP id l49KgD7T016786 } 9, 19 -- for {microscopy-at-microscopy.com} ; Wed, 9 May 2007 14:42:13 -0600 } 9, 19 -- From: "John Chandler" {jpchandl-at-mines.edu} } 9, 19 -- To: {microscopy-at-microscopy.com} } 9, 19 -- Subject: Looking for recommendations about dual beam ion mills } 9, 19 -- Date: Wed, 9 May 2007 14:42:13 -0600 } 9, 19 -- Message-ID: {001e01c7927a$861c8570$ad1c438a-at-mines.edu} } 9, 19 -- MIME-Version: 1.0 } 9, 19 -- Content-Type: text/plain; } 9, 19 -- charset="us-ascii" } 9, 19 -- Content-Transfer-Encoding: 7bit } 9, 19 -- X-Mailer: Microsoft Office Outlook 11 } 9, 19 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 } 9, 19 -- Thread-Index: AceSeoX7iyPEBSG3T3O4yAVZe0waUw== } ==============================End of - Headers==============================
==============================Original Headers============================== 11, 24 -- From nealzimm-at-cpinternet.com Thu May 10 08:08:28 2007 11, 24 -- Received: from cpinternet.com (mail.cpinternet.com [209.240.224.81]) 11, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4AD8S9W001596 11, 24 -- for {microscopy-at-microscopy.com} ; Thu, 10 May 2007 08:08:28 -0500 11, 24 -- Received: from [192.168.1.102] (unverified [216.251.191.121]) 11, 24 -- by cpinternet.com (SurgeMail 3.8j) with ESMTP id 171217060-1881964 11, 24 -- for multiple; Thu, 10 May 2007 08:08:27 -0500 CDT 11, 24 -- Subject: Re: [Microscopy] Looking for recommendations about dual beam ion 11, 24 -- mills 11, 24 -- From: Neal Zimmermann {nealzimm-at-cpinternet.com} 11, 24 -- To: jpchandl-at-mines.edu, microscopy-at-microscopy.com 11, 24 -- In-Reply-To: {200705092043.l49Kh2W8005876-at-ns.microscopy.com} 11, 24 -- References: {200705092043.l49Kh2W8005876-at-ns.microscopy.com} 11, 24 -- Content-Type: text/plain 11, 24 -- Date: Thu, 10 May 2007 08:08:14 -0500 11, 24 -- Message-Id: {1178802494.3663.19.camel-at-localhost.localdomain} 11, 24 -- Mime-Version: 1.0 11, 24 -- X-Mailer: Evolution 2.8.3 (2.8.3-2.fc6) 11, 24 -- Content-Transfer-Encoding: 7bit 11, 24 -- X-Authenticated-User: nealzimm-at-cpinternet.com 11, 24 -- X-NotAscii: charset=us-ascii 11, 24 -- X-Encryption: SSL encrypted 11, 24 -- X-IP-stats: No info recorded yet ip=216.251.191.121 11, 24 -- X-Originating-IP: 216.251.191.121 ==============================End of - Headers==============================
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==============================Original Headers============================== 23, 34 -- From richard.beanland-at-bookham.com Thu May 10 09:09:28 2007 23, 34 -- Received: from mail72.messagelabs.com (mail72.messagelabs.com [193.109.255.147]) 23, 34 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l4AE9Riq025754 23, 34 -- for {microscopy-at-microscopy.com} ; Thu, 10 May 2007 09:09:27 -0500 23, 34 -- X-VirusChecked: Checked 23, 34 -- X-Env-Sender: richard.beanland-at-bookham.com 23, 34 -- X-Msg-Ref: server-10.tower-72.messagelabs.com!1178806165!33452822!1 23, 34 -- X-StarScan-Version: 5.5.10.7.1; banners=bookham.com,-,- 23, 34 -- X-Originating-IP: [213.249.209.179] 23, 34 -- Received: (qmail 31853 invoked from network); 10 May 2007 14:09:26 -0000 23, 34 -- Received: from unknown (HELO cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI) (213.249.209.179) 23, 34 -- by server-10.tower-72.messagelabs.com with SMTP; 10 May 2007 14:09:26 -0000 23, 34 -- Received: from cas-smx-02.BOOKHAM.ENTERPRISE.PRI ([10.4.0.223]) by cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI with Microsoft SMTPSVC(6.0.3790.1830); 23, 34 -- Thu, 10 May 2007 15:10:39 +0100 23, 34 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 23, 34 -- Content-class: urn:content-classes:message 23, 34 -- MIME-Version: 1.0 23, 34 -- Content-Type: text/plain; 23, 34 -- charset="us-ascii" 23, 34 -- Subject: RE: [Microscopy] Re: Looking for recommendations about dual beam ion 23, 34 -- Date: Thu, 10 May 2007 15:10:38 +0100 23, 34 -- Message-ID: {9645D3E33E4C6548B12A7B25F611533E46C9B1-at-cas-smx-02.BOOKHAM.ENTERPRISE.PRI} 23, 34 -- In-Reply-To: {200705101309.l4AD9WRs003477-at-ns.microscopy.com} 23, 34 -- X-MS-Has-Attach: 23, 34 -- X-MS-TNEF-Correlator: 23, 34 -- Thread-Topic: [Microscopy] Re: Looking for recommendations about dual beam ion 23, 34 -- Thread-Index: AceTBKSYNfWLTtdMSI21epLGVDAwXgABIZoA 23, 34 -- References: {200705101309.l4AD9WRs003477-at-ns.microscopy.com} 23, 34 -- From: "Richard Beanland" {richard.beanland-at-bookham.com} 23, 34 -- To: {nealzimm-at-cpinternet.com} 23, 34 -- Cc: {microscopy-at-microscopy.com} 23, 34 -- X-OriginalArrivalTime: 10 May 2007 14:10:39.0130 (UTC) FILETIME=[FCAAEFA0:01C7930C] 23, 34 -- Content-Transfer-Encoding: 8bit 23, 34 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4AE9Riq025754 ==============================End of - Headers==============================
Dear all, I heard some people to use a solution containing citric acid and H2O2 to remove the damage during ion milling. Does anyone have experience with this? What kind of damage it can remove, amorphorized pits, deposited contamination? Does it require a following cleaning process? Can someone with experience kindly give a detailed description of this technique? Thanks
Shu Miao
==============================Original Headers============================== 2, 21 -- From shu-at-caltech.edu Thu May 10 09:52:34 2007 2, 21 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 2, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4AEqXQ1005732 2, 21 -- for {Microscopy-at-microscopy.com} ; Thu, 10 May 2007 09:52:34 -0500 2, 21 -- Received: from fire-dog.its.caltech.edu (fire-dog [192.168.1.4]) 2, 21 -- by fire-ox-postvirus (Postfix) with ESMTP id 575F0352F3 2, 21 -- for {Microscopy-at-microscopy.com} ; Thu, 10 May 2007 07:52:33 -0700 (PDT) 2, 21 -- Received: from sue (sue.its.caltech.edu [131.215.239.44]) 2, 21 -- (using TLSv1 with cipher EDH-RSA-DES-CBC3-SHA (168/168 bits)) 2, 21 -- (No client certificate requested) 2, 21 -- by fire-ox.its.caltech.edu (Postfix) with ESMTP id 32A0A3531E 2, 21 -- for {Microscopy-at-microscopy.com} ; Thu, 10 May 2007 07:52:32 -0700 (PDT) 2, 21 -- Date: Thu, 10 May 2007 07:52:31 -0700 (PDT) 2, 21 -- From: Shu Miao {shu-at-caltech.edu} 2, 21 -- X-X-Sender: shu-at-sue 2, 21 -- To: Microscopy-at-microscopy.com 2, 21 -- Subject: Etching of ion milled sample 2, 21 -- Message-ID: {Pine.GSO.4.58.0705100744230.14730-at-sue} 2, 21 -- MIME-Version: 1.0 2, 21 -- Content-Type: TEXT/PLAIN; charset=US-ASCII 2, 21 -- X-Spam-Scanned: at Caltech-ITS on fire-dog by amavisd-2.4.5 ==============================End of - Headers==============================
"I've tried a number of other things for getting epoxy semi-thins to stick to glass, "
You didn't state how thick nor what area your sections are, but for me semi-thins are nominally 2.0 microns thick and usually no larger than 4 mm per side
I use no slide subbing method. I clean 1x3" glass slides with an ethanol rinse, then air dry at room temperature or blow down with hair dryer.
I then collect sections on a drop or two of distilled water on the slide, transfered there from the microtome with a clean fine tipped artists brush. I collect about 8-12 sections per drop.
Then I warm the slide beneath the water drop from below using an alcohol lamp, fairly hot, but not to boil, of course. After drying by heating the sections stick quite well. I can then stain, usually with 0.2 micron filtered toluidine blue, again heating but gently this time, for about a minute, until stain "develops" the section. Then rinse that stain off with distilled water from a squirt bottle, even direct the spray right onto the sections to get rid of any ppt. Dry again gently with flame - then view on the microscope.
If you are cutting much thicker sections, say 10 micron or more, or sections have much larger area, maybe this would not work, tho I have not tried this method for those larger sections.
Hope this helps, but I have a sneaky feeling that I've missed the point of what you are trying to do and that some kind of subbing is quite necessary for accomplishing your goals. Anyway, thought I'd toss this out for what its worth.
Gib -- Gilbert (Gib) Ahlstrand, Electron Microscopist, Imaging Center, University of Minnesota 123 Snyder Hall St. Paul, MN 55108 http://www.cbs.umn.edu/ic
dac-at-research.umass.edu wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Hi everyone, } } I'm making subbed slides and just wondering if there is anyone who knows } the chemistry/function of the chromium potassium sulfate (chrome alum) } in the mix, and if there is a new-age environmentally-friendly } substitute for it, or can it be eliminated - to what effect? I'm working } on the assumption that it has a function. I know that only small } quantities of Chrome alum are used, but would like to eliminate it if } possible. } } To avoid too much traffic about alternatives, let me say I've tried a } number of other things for getting epoxy semi-thins to stick to glass, } and this method works for me. I did already see a note in the archives } about a "pinch of gelatine in the water bath" and that doesn't have the } chromium compound. I've also read an alternative use of the Mayer's egg } albumin - instead of smearing the slide and drying, adding some to the } flotation water - and that also doesn't contain chromium. I like the way } water drops sit nicely on the subbed slide so rows of sequential } sections can be arrayed in the droplets without mixing. } } } Thanks in advance for any insights. } } } Dale Callaham }
==============================Original Headers============================== 11, 21 -- From ahlst007-at-umn.edu Thu May 10 10:11:06 2007 11, 21 -- Received: from mta-w2.tc.umn.edu (mta-w2.tc.umn.edu [134.84.119.6]) 11, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4AFB65o017391 11, 21 -- for {Microscopy-at-Microscopy.com} ; Thu, 10 May 2007 10:11:06 -0500 11, 21 -- Received: from [160.94.39.28] (x160-94-39-28.cbs.umn.edu [160.94.39.28]) 11, 21 -- by mta-w2.tc.umn.edu (UMN smtpd) with ESMTP 11, 21 -- for {Microscopy-at-Microscopy.com} ; Thu, 10 May 2007 10:10:26 -0500 (CDT) 11, 21 -- X-Umn-Remote-Mta: [N] x160-94-39-28.cbs.umn.edu [160.94.39.28] #+LO+TS+AU+HN 11, 21 -- Message-ID: {46433548.9020406-at-umn.edu} 11, 21 -- Date: Thu, 10 May 2007 10:07:52 -0500 11, 21 -- From: Gib Ahlstrand {ahlst007-at-umn.edu} 11, 21 -- Reply-To: ahlst007-at-umn.edu 11, 21 -- Organization: Imaging Center UM 11, 21 -- User-Agent: Thunderbird 1.5.0.10 (Macintosh/20070221) 11, 21 -- MIME-Version: 1.0 11, 21 -- To: Microscopy-at-Microscopy.com 11, 21 -- Subject: Re: Gelatin subbed slides - eliminate the chromium? 11, 21 -- References: {200705092033.l49KXnAh030483-at-ns.microscopy.com} 11, 21 -- In-Reply-To: {200705092033.l49KXnAh030483-at-ns.microscopy.com} 11, 21 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 11, 21 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
I am so glad to see the discussion on the contrast issue continues as I have been hearing more and more reports on this problem and I myself have been experiencing it too. Many hypotheses have been suggested on the causes of the problem but it seems that there were always evidences supporting different arguments. Practically, besides using short dehydration and infiltration times for monolayer cells, I now HAVE TO use potassium ferrocyanide with OsO4 in order to get good contrast (even for bulk tissue). When I find out the contrast is low after sections are being cut, I re-counterstain sections with 15% UA in methanol (it can give a muddy appearance if it is over done). In addition, I make sure to use a relatively fresh OsO4 stock solution.
I have been working in EM core facilities for a long time and have dealt with all kinds of samples. One question I have for people like me is whether this is a new problem or has always been this way. I did not remember contrast was so much an issue in the good old days even with standard dehydration times and when OsO4 was used alone. Personally I think this problem has something to do with the reagents we use now days, particularly OsO4. I did notice the results were different when I use different OsO4 (liquid vs. crystal, old vs. fresh….). I am wondering what other people’s experiences are with OsO4 and how OsO4 solution is prepared and stored in different laboratories. I would really love to hear from chemist more details on how OsO4 works and from vendors if there has been any change in their sources for OsO4.
Thank all very much
Hong Emory SOM EM
On May 10, 2007, at 8:10 AM, malcolm.haswell-at-sunderland.ac.uk wrote: } } ----------------------------------------------------------------------- } ----- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } ----- } } Dear Tom } } I just thought I should mention something that I think I read years ago } in one of M.A. Hayat's books. He mentions that cacodylate and some } other } buffers when incorporated into a fixative produce higher contrast } because they are more extractive whereas phosphate buffers probably } preserve cell contents better but at the expense of contrast. } } I just thought that I should add this because you mention Sorenson's } buffer in the fixative. I'm sure that many of the other points have an } effect but just wondered if the buffer could be contributing to your } problems as well. } } Malcolm } } } Malcolm Haswell } e.m. unit } School of Health, Natural and Social Sciences } Fleming Building } University of Sunderland } Tyne & Wear } SR1 3SD } UK } e-mail: malcolm.haswell-at-sunderland.ac.uk } } } ----- Original Message ----- } X-from: nizets2-at-yahoo.com } Date: Thursday, May 10, 2007 12:23 pm } Subject: [Microscopy] Re: Help with enhancing contrast of Mitochondrial } cristae } } } } } } } } } -------------------------------------------------------------------- } } -------- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } } AmericaTo Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserverOn-Line Help } } http://www.microscopy.com/MicroscopyListserver/FAQ.html------------- } } --------------------------------------------------------------- } } } } Dear Tom, } } } } I agree with the other comments: Ferrocyanide, } } decreasing dehydration times (penetration is immediate } } in monolayers) and using picric acid probably can } } improve membrane contrast. } } (personally I already tried UAc in methanol without } } satisfaction) } } } } Please share your experience with us. } } } } Best regards, } } } } Stephane } } } } --- tbargar-at-unmc.edu wrote: } } } } } } } } } } } } } } } } -------------------------------------------------------------------- } } -------- } } } The Microscopy ListServer -- CoSponsor: The } } } Microscopy Society of America } } } To Subscribe/Unsubscribe -- } } } http://www.microscopy.com/MicroscopyListserver } } } On-Line Help } } } } } http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } } } -------------------------------------------------------------------- } } -------- } } } } } } } } } Dear listers, } } } } } } Anyone out there have any advice on how to enhance } } } the contrast and } } } definition of the membranes of the cristae of } } } mitochondria? The samples } } } brought to me are monolayer cell cultures of cancer } } } cells grown on } } } Thermanox coverslips. This is how I'm currently } } } processing the samples: } } } Primary fixation is 2% glutaraldehyde, 2% } } } paraformaldehyde, 0.5% acrolein } } } in 0.1M Sorensen's phosphate buffer pH 7.2. } } } Post-fixation in 1%Osmium } } } Tetroxide in 0.1M Sorensen's phosphate buffer. } } } Dehydration in 50, 70, 90, } } } 95, 100%X3 ethanol solutions. embedding in } } } Araldite. Sections are stained } } } with 2% uranyl acetate aqueous 15 minutes and } } } Reynold's lead citrate 10 } } } minutes. The density of the cytoplasm and } } } mitochondrial matrix are } } } similar with the result that the contrast of the } } } mitochondria is similar to } } } the cytoplasm. The mitochondria and it's membranes } } } (outer and that of the } } } cristae) don't really "stand out". The researchers } } } involved want to see } } } really contrasty mitochondrial cristae. } } } } } } The next thing I'm going to try is post-fixation } } } with a mix of osmium } } } tetroxide and potassium ferrocyanide. } } } } } } Anyone have any other ideas? Thanks in advance. } } } } } } } } } Tom Bargar } } } University of Nebraska Medical Center } } } Core Electron Microscopy Research Facility } } } 986395 Nebraska Medical Center } } } Omaha, NE 68198-6395 } } } 402-559-7347 } } } tbargar-at-unmc.edu } } } } } } } } } ==============================Original } } } Headers============================== } } } 8, 20 -- From tbargar-at-unmc.edu Tue May 1 15:39:39 } } } 2007 } } } 8, 20 -- Received: from zixvpm02.unmc.edu } } } (zixvpm02.unmc.edu [192.198.54.127]) } } } 8, 20 -- by ns.microscopy.com } } } (8.12.11.20060308/8.12.8) with ESMTP id } } } l41KddQH023357 } } } 8, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, } } } 1 May 2007 15:39:39 -0500 } } } 8, 20 -- Received: from zixvpm02.unmc.edu (ZixVPM } } } [127.0.0.1]) } } } 8, 20 -- by Outbound.unmc.edu (Proprietary) with } } } ESMTP id 8A304A0061 } } } 8, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, } } } 1 May 2007 15:39:39 -0500 (CDT) } } } 8, 20 -- Received: from unmcnotes01.unmc.edu } } } (host-137-197-103-35.unmc.edu [137.197.103.35]) } } } 8, 20 -- by zixvpm02.unmc.edu (Proprietary) with } } } ESMTP id 72E1039804A } } } 8, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, } } } 1 May 2007 15:39:38 -0500 (CDT) } } } 8, 20 -- Subject: Help with enhancing contrast of } } } Mitochondrial cristae } } } 8, 20 -- To: Microscopy-at-MSA.Microscopy.Com } } } 8, 20 -- X-Mailer: Lotus Notes Release 7.0.1 January } } } 17, 2006 } } } 8, 20 -- Message-ID: } } } } } {OFDC8670F1.FEB83264-ON862572CE.006F5771-862572CE.00717D6E-at-unmc.edu} } } } 8, 20 -- From: Tom W Bargar {tbargar-at-unmc.edu} } } } 8, 20 -- Date: Tue, 1 May 2007 15:39:27 -0500 } } } 8, 20 -- X-MIMETrack: Serialize by Router on } } } UNMCNOTES01.UNMC.EDU/Servers/UNEBR at 05/01/2007 } } } 03:39:38 } } } 8, 20 -- PM } } } 8, 20 -- MIME-Version: 1.0 } } } 8, 20 -- Content-type: text/plain; charset=US-ASCII } } } ==============================End of - } } } Headers============================== } } } } } } } } } __________________________________________________ } } Do You Yahoo!? } } Tired of spam? 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Mail has the best spam protection around } } http://mail.yahoo.com } } } } ==============================Original } } Headers==============================9, 21 -- From } } nizets2-at-yahoo.com Thu May 10 06:15:21 2007 } } 9, 21 -- Received: from web37402.mail.mud.yahoo.com } } (web37402.mail.mud.yahoo.com [209.191.91.134]) } } 9, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP } } id l4ABFKJw009245 } } 9, 21 -- for {microscopy-at-microscopy.com} ; Thu, 10 May 2007 } } 06:15:21 -0500 } } 9, 21 -- Received: (qmail 27735 invoked by uid 60001); 10 May 2007 } } 11:15:20 -0000 } } 9, 21 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; } } 9, 21 -- s=s1024; d=yahoo.com; } } 9, 21 -- h=X-YMail-OSG:Received:Date:From:Subject:To:Cc:In-Reply- } } To:MIME-Version:Content-Type:Content-Transfer-Encoding:Message-ID; } } 9, 21 -- } } } b=xdVFiLCF4Y+sO8r3CpubJCf2uWbS/tsa8x0ab/ } hPl+5HFUOENjJ6yIvxMiFISE4+rsVrEo+9g9amUFa4qIQOAGV0FBzymuaE0sNVDp0SglZlj } Ly8sAz3+WIuaj+c/D2DZnTf4n+Vivh1RJ9XqSQvZy3HvK1FSpB028byJ8YI300=;9, } 21 -- X-YMail-OSG: } mktHeDIVM1kftlmU8fPc3P3OX17YmyvCJT352ac98PdSr0qhC7X.fCqkPbxI6mkxa09GcWQ } MmEhQCWAHO8TzXOi4W.ueJ.8eBX54nP.Y7I3f9c1fcF6eyrST0RA3CoDbTEtqWbHpql.Ve3 } 3A65i7tH.ZVhYJ5Jn8TfeDlO7pDP4W.DeEznuXRaE- } } 9, 21 -- Received: from [80.122.101.102] by } } web37402.mail.mud.yahoo.com via HTTP; Thu, 10 May 2007 04:15:20 PDT } } 9, 21 -- Date: Thu, 10 May 2007 04:15:20 -0700 (PDT) } } 9, 21 -- From: Stephane Nizet {nizets2-at-yahoo.com} } } 9, 21 -- Subject: Re: [Microscopy] Help with enhancing contrast of } } Mitochondrial cristae } } 9, 21 -- To: tbargar-at-unmc.edu } } 9, 21 -- Cc: microscopy-at-microscopy.com } } 9, 21 -- In-Reply-To: {200705012045.l41Kjoma031136-at-ns.microscopy.com} } } 9, 21 -- MIME-Version: 1.0 } } 9, 21 -- Content-Type: text/plain; charset=iso-8859-1 } } 9, 21 -- Content-Transfer-Encoding: 8bit } } 9, 21 -- Message-ID: {635982.26761.qm-at-web37402.mail.mud.yahoo.com} } } ==============================End of - } } Headers============================== } } ==============================Original } Headers============================== } 8, 37 -- From malcolm.haswell-at-sunderland.ac.uk Thu May 10 07:06:52 2007 } 8, 37 -- Received: from max1.sunderland.ac.uk (max1.sunderland.ac.uk } [157.228.29.83]) } 8, 37 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id } l4AC6os7021493 } 8, 37 -- for {Microscopy-at-microscopy.com} ; Thu, 10 May 2007 07:06:51 } -0500 } 8, 37 -- Received: (qmail 1859 invoked from network); 10 May 2007 } 12:06:45 -0000 } 8, 37 -- Received: from localhost (127.0.0.1) } 8, 37 -- by max1.sunderland.ac.uk with SMTP; 10 May 2007 12:06:45 } -0000 } 8, 37 -- Received: from max1.sunderland.ac.uk ([127.0.0.1]) } 8, 37 -- by localhost (max1.sunderland.ac.uk [127.0.0.1]) } (amavisd-new, port 10024) } 8, 37 -- with SMTP id 25620-10 for {Microscopy-at-microscopy.com} ; } 8, 37 -- Thu, 10 May 2007 13:06:38 +0100 (BST) } 8, 37 -- Received: (qmail 1200 invoked by uid 607); 10 May 2007 } 12:06:34 -0000 } 8, 37 -- Received: from [157.228.37.117] (HELO } hermes.sunderland.ac.uk) (157.228.37.117) } 8, 37 -- by max1.sunderland.ac.uk (qpsmtpd/0.28) with ESMTP; Thu, } 10 May 2007 13:06:34 +0100 } 8, 37 -- Received: from sunderland.ac.uk (localhost [127.0.0.1]) } 8, 37 -- by hermes.sunderland.ac.uk } 8, 37 -- (iPlanet Messaging Server 5.2 Patch 2 (built Jul 14 2004)) } 8, 37 -- with ESMTP id {0JHT00GFAQZ00H-at-hermes.sunderland.ac.uk} for } 8, 37 -- Microscopy-at-microscopy.com; Thu, 10 May 2007 13:06:36 +0100 } (BST) } 8, 37 -- Received: from [157.228.75.11] by hermes.sunderland.ac.uk } (mshttpd); Thu, } 8, 37 -- 10 May 2007 13:06:36 +0100 } 8, 37 -- Date: Thu, 10 May 2007 13:06:36 +0100 } 8, 37 -- From: Malcolm Haswell {malcolm.haswell-at-sunderland.ac.uk} } 8, 37 -- Subject: Re: [Microscopy] Re: Help with enhancing contrast of } Mitochondrial } 8, 37 -- cristae } 8, 37 -- To: Microscopy MSA {Microscopy-at-microscopy.com} } 8, 37 -- Cc: tbargar-at-unmc.edu } 8, 37 -- Message-id: {135574d1354105.1354105135574d-at-sunderland.ac.uk} } 8, 37 -- MIME-version: 1.0 } 8, 37 -- X-Mailer: iPlanet Messenger Express 5.2 Patch 2 (built Jul 14 } 2004) } 8, 37 -- Content-type: text/plain; charset=us-ascii } 8, 37 -- Content-language: en } 8, 37 -- Content-transfer-encoding: 7BIT } 8, 37 -- Content-disposition: inline } 8, 37 -- X-Accept-Language: en } 8, 37 -- Priority: normal } 8, 37 -- X-Virus-Scanned: by iCritical at max1.sunderland.ac.uk } ==============================End of - } Headers==============================
==============================Original Headers============================== 9, 27 -- From hyi-at-emory.edu Thu May 10 10:40:47 2007 9, 27 -- Received: from virginia.cc.emory.edu (virginia.cc.emory.edu [170.140.8.222]) 9, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4AFelNb029509 9, 27 -- for {microscopy-at-microscopy.com} ; Thu, 10 May 2007 10:40:47 -0500 9, 27 -- Received: from virginia (virginia [170.140.8.222]) 9, 27 -- by virginia.cc.emory.edu (8.13.8/8.13.8) with ESMTP id l4AFekQN025533 9, 27 -- for {microscopy-at-microscopy.com} ; Thu, 10 May 2007 11:40:46 -0400 (EDT) 9, 27 -- Received: from virginia (unknown [127.0.0.1]) 9, 27 -- by virginia (Symantec Mail Security) with ESMTP id 8C3CB24FB 9, 27 -- for {microscopy-at-microscopy.com} ; Thu, 10 May 2007 11:40:46 -0400 (EDT) 9, 27 -- X-AuditID: aa8c08de-00000005000004ef-d9-46433cfd4036 9, 27 -- Received: from [170.140.233.63] (unknown [170.140.233.63]) 9, 27 -- by virginia (Symantec Mail Security) with ESMTP id C50A924FC 9, 27 -- for {microscopy-at-microscopy.com} ; Thu, 10 May 2007 11:40:45 -0400 (EDT) 9, 27 -- Mime-Version: 1.0 (Apple Message framework v622) 9, 27 -- In-Reply-To: {200705101210.l4ACA3fM026200-at-ns.microscopy.com} 9, 27 -- References: {200705101210.l4ACA3fM026200-at-ns.microscopy.com} 9, 27 -- Content-Type: text/plain; charset=WINDOWS-1252; delsp=yes; format=flowed 9, 27 -- Message-Id: {8c61afeb0634c079641ccf803afaa602-at-emory.edu} 9, 27 -- From: Hong Yi {hyi-at-emory.edu} 9, 27 -- Subject: [Microscopy] Help with enhancing contrast of Mitochondrial 9, 27 -- Date: Thu, 10 May 2007 12:40:07 -0400 9, 27 -- To: microscopy-at-microscopy.com 9, 27 -- X-Mailer: Apple Mail (2.622) 9, 27 -- X-Brightmail-Tracker: AAAAAA== 9, 27 -- Content-Transfer-Encoding: 8bit 9, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4AFelNb029509 ==============================End of - Headers==============================
More osmium questions: I still think that a lot of the trouble with low-contrast ("missing") membranes in TC samples is due to the containers - if you run a cell pellet (in a polypropylene Eppendorf tube) side-by-side with cells in their culture dishes (polystyrene), the pelleted cells will look normal, but the dish cells will have the "ghost" membranes....and the PP tube is still clear, but the PS dish has browned a bit after osmication. Using a reduced osmium (OPF) avoids the problem. I can't figure out why the post treatments some people use reverse this, and I've never tried those - I use OPF for TC samples, but plan to give those a whirl in my free time (maybe when I retire?!).
Any chemists able to hop in on the container issue? Please?
Tamara
On Thu, 10 May 2007 10:42:01 -0500 hyi-at-emory.edu wrote: } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy } Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Dear All: } } I am so glad to see the discussion on the } contrast issue } continues as I have been hearing more and more reports } on this problem } and I myself have been experiencing it too. Many } hypotheses have been } suggested on the causes of the problem but it seems that } there were } always evidences supporting different } arguments. Practically, besides } using short dehydration and infiltration times for } monolayer cells, I } now HAVE TO use potassium ferrocyanide with OsO4 in } order to get good } contrast (even for bulk tissue). When I find out the } contrast is low } after sections are being cut, I re-counterstain sections } with 15% UA in } methanol (it can give a muddy appearance if it is over } done). In } addition, I make sure to use a relatively fresh OsO4 } stock solution. } } I have been working in EM core facilities for a } long time and } have dealt with all kinds of samples. One question I } have for people } like me is whether this is a new problem or has always } been this way. I } did not remember contrast was so much an issue in the } good old days } even with standard dehydration times and when OsO4 was } used } alone. Personally I think this problem has something to } do with the } reagents we use now days, particularly OsO4. I did } notice the results } were different when I use different OsO4 (liquid vs. } crystal, old vs. } fresh….). I am wondering what other people’s experiences } are with OsO4 } and how OsO4 solution is prepared and stored in } different } laboratories. I would really love to hear from chemist } more details on } how OsO4 works and from vendors if there has been any } change in their } sources for OsO4. } } Thank all very much } } Hong } Emory SOM EM } } } On May 10, 2007, at 8:10 AM, } malcolm.haswell-at-sunderland.ac.uk wrote: } } } } ----------------------------------------------------------------------- } } ----- } } The Microscopy ListServer -- CoSponsor: The Microscopy } } Society of } } America } } To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver } } On-Line Help } } http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ----------------------------------------------------------------------- } } ----- } } } } Dear Tom } } } } I just thought I should mention something that I think I } } read years ago } } in one of M.A. Hayat's books. He mentions that } } cacodylate and some } } other } } buffers when incorporated into a fixative produce higher } } contrast } } because they are more extractive whereas phosphate } } buffers probably } } preserve cell contents better but at the expense of } } contrast. } } } } I just thought that I should add this because you } } mention Sorenson's } } buffer in the fixative. I'm sure that many of the other } } points have an } } effect but just wondered if the buffer could be } } contributing to your } } problems as well. } } } } Malcolm } } } } } } Malcolm Haswell } } e.m. unit } } School of Health, Natural and Social Sciences } } Fleming Building } } University of Sunderland } } Tyne & Wear } } SR1 3SD } } UK } } e-mail: malcolm.haswell-at-sunderland.ac.uk } } } } } } ----- Original Message ----- } } X-from: nizets2-at-yahoo.com } } Date: Thursday, May 10, 2007 12:23 pm } } Subject: [Microscopy] Re: Help with enhancing contrast } } of Mitochondrial } } cristae } } } } } } } } } } } } } } -------------------------------------------------------------------- } } } -------- } } } The Microscopy ListServer -- CoSponsor: The Microscopy } } } Society of } } } AmericaTo Subscribe/Unsubscribe -- } } } http://www.microscopy.com/MicroscopyListserverOn-Line } } } Help } } } http://www.microscopy.com/MicroscopyListserver/FAQ.html------------- } } } --------------------------------------------------------------- } } } } } } Dear Tom, } } } } } } I agree with the other comments: Ferrocyanide, } } } decreasing dehydration times (penetration is immediate } } } in monolayers) and using picric acid probably can } } } improve membrane contrast. } } } (personally I already tried UAc in methanol without } } } satisfaction) } } } } } } Please share your experience with us. } } } } } } Best regards, } } } } } } Stephane } } } } } } --- tbargar-at-unmc.edu wrote: } } } } } } } } } } } } } } } } } } } } } } -------------------------------------------------------------------- } } } -------- } } } } The Microscopy ListServer -- CoSponsor: The } } } } Microscopy Society of America } } } } To Subscribe/Unsubscribe -- } } } } http://www.microscopy.com/MicroscopyListserver } } } } On-Line Help } } } } } } } http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } } -------------------------------------------------------------------- } } } -------- } } } } } } } } } } } } Dear listers, } } } } } } } } Anyone out there have any advice on how to enhance } } } } the contrast and } } } } definition of the membranes of the cristae of } } } } mitochondria? The samples } } } } brought to me are monolayer cell cultures of cancer } } } } cells grown on } } } } Thermanox coverslips. This is how I'm currently } } } } processing the samples: } } } } Primary fixation is 2% glutaraldehyde, 2% } } } } paraformaldehyde, 0.5% acrolein } } } } in 0.1M Sorensen's phosphate buffer pH 7.2. } } } } Post-fixation in 1%Osmium } } } } Tetroxide in 0.1M Sorensen's phosphate buffer. } } } } Dehydration in 50, 70, 90, } } } } 95, 100%X3 ethanol solutions. embedding in } } } } Araldite. Sections are stained } } } } with 2% uranyl acetate aqueous 15 minutes and } } } } Reynold's lead citrate 10 } } } } minutes. The density of the cytoplasm and } } } } mitochondrial matrix are } } } } similar with the result that the contrast of the } } } } mitochondria is similar to } } } } the cytoplasm. The mitochondria and it's membranes } } } } (outer and that of the } } } } cristae) don't really "stand out". The researchers } } } } involved want to see } } } } really contrasty mitochondrial cristae. } } } } } } } } The next thing I'm going to try is post-fixation } } } } with a mix of osmium } } } } tetroxide and potassium ferrocyanide. } } } } } } } } Anyone have any other ideas? Thanks in advance. } } } } } } } } } } } } Tom Bargar } } } } University of Nebraska Medical Center } } } } Core Electron Microscopy Research Facility } } } } 986395 Nebraska Medical Center } } } } Omaha, NE 68198-6395 } } } } 402-559-7347 } } } } tbargar-at-unmc.edu } } } } } } } } } } } } ==============================Original } } } } Headers============================== } } } } 8, 20 -- From tbargar-at-unmc.edu Tue May 1 15:39:39 } } } } 2007 } } } } 8, 20 -- Received: from zixvpm02.unmc.edu } } } } (zixvpm02.unmc.edu [192.198.54.127]) } } } } 8, 20 -- by ns.microscopy.com } } } } (8.12.11.20060308/8.12.8) with ESMTP id } } } } l41KddQH023357 } } } } 8, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, } } } } 1 May 2007 15:39:39 -0500 } } } } 8, 20 -- Received: from zixvpm02.unmc.edu (ZixVPM } } } } [127.0.0.1]) } } } } 8, 20 -- by Outbound.unmc.edu (Proprietary) with } } } } ESMTP id 8A304A0061 } } } } 8, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, } } } } 1 May 2007 15:39:39 -0500 (CDT) } } } } 8, 20 -- Received: from unmcnotes01.unmc.edu } } } } (host-137-197-103-35.unmc.edu [137.197.103.35]) } } } } 8, 20 -- by zixvpm02.unmc.edu (Proprietary) with } } } } ESMTP id 72E1039804A } } } } 8, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, } } } } 1 May 2007 15:39:38 -0500 (CDT) } } } } 8, 20 -- Subject: Help with enhancing contrast of } } } } Mitochondrial cristae } } } } 8, 20 -- To: Microscopy-at-MSA.Microscopy.Com } } } } 8, 20 -- X-Mailer: Lotus Notes Release 7.0.1 January } } } } 17, 2006 } } } } 8, 20 -- Message-ID: } } } } } } } {OFDC8670F1.FEB83264-ON862572CE.006F5771-862572CE.00717D6E-at-unmc.edu} } } } } 8, 20 -- From: Tom W Bargar {tbargar-at-unmc.edu} } } } } 8, 20 -- Date: Tue, 1 May 2007 15:39:27 -0500 } } } } 8, 20 -- X-MIMETrack: Serialize by Router on } } } } UNMCNOTES01.UNMC.EDU/Servers/UNEBR at 05/01/2007 } } } } 03:39:38 } } } } 8, 20 -- PM } } } } 8, 20 -- MIME-Version: 1.0 } } } } 8, 20 -- Content-type: text/plain; charset=US-ASCII } } } } ==============================End of - } } } } Headers============================== } } } } } } } } } } } } } __________________________________________________ } } } Do You Yahoo!? } } } Tired of spam? Yahoo! Mail has the best spam protection } } } around } } } http://mail.yahoo.com } } } } } } ==============================Original } } } Headers==============================9, 21 -- From } } } nizets2-at-yahoo.com Thu May 10 06:15:21 2007 } } } 9, 21 -- Received: from web37402.mail.mud.yahoo.com } } } (web37402.mail.mud.yahoo.com [209.191.91.134]) } } } 9, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8)
This message has bounced from the listserver several times. I am trying again but have deleted the protocol to see if this is the cause.
Yes, I am familiar with the older method requiring dry acetone. But the half life of APTS in water pH 7 at 24 C is 8.4 hrs (see nice summary of chemical properties and safety at http://www.inchem.org/documents/sids/sids/919302.pdf ). So my advice is make it fresh and skip the hassles with acetone. This means you can use plastic slide holders to dip!
At 05:16 PM 05/09/07, you wrote: } Hi Tom, } } Ok, I'm interested... I have used 3-aminopropyltriethoxysilane (same } thing?) at 1% in acetone to treat glass so that things with aldehydes } would stick, but the notes I had on that (method from Hans Ris) indicate } absolutely dry acetone - that water would decompose it. So you mix it with } water? } } Thanks, } } Dale } } Thomas E. Phillips wrote: } } If you haven't tried coating your slides with 1% } } aminopropyltriethoxysilane (APTS) in water then you are missing out on a } } much better alternative. The water really beads up on it. I used to use } } chrome-gel but it doesn't compare. } } } } At 03:30 PM 05/09/07, you wrote: } } } } } } } ---------------------------------------------------------------------------- } } } } } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
Thank you, Tamara. I have seen what you described as well. I even lifted cells from half of culture dish and processed it in parallel (different resin though) in a Eppendorf tube. The contrast of cells in tube was somewhat better than those from the dish. However, I am puzzled by the fact that we process brain vibratome sections in dishes routinely and have not had much problem with contrast. If the argument for this is that the vibratome sections are not attached to the plastic, then so is monolayer cells on glass coverslip. I have seen low contrast in cells cultured on glass coverslip as well.
Nevertheless, I do think container could be a possible cause, and do want to hear from explanation from a chemist.
Hong
On May 10, 2007, at 12:01 PM, Tamara A Howard wrote:
} More osmium questions: I still think that a lot of the trouble with } low-contrast ("missing") membranes in TC samples is due to the } containers - if you run a cell pellet (in a polypropylene Eppendorf } tube) side-by-side with cells in their culture dishes (polystyrene), } the pelleted cells will look normal, but the dish cells will have the } "ghost" membranes....and the PP tube is still clear, but the PS dish } has browned a bit after osmication. Using a reduced osmium (OPF) } avoids the problem. I can't figure out why the post treatments some } people use reverse this, and I've never tried those - I use OPF for TC } samples, but plan to give those a whirl in my free time (maybe when I } retire?!). } } Any chemists able to hop in on the container issue? Please? } } Tamara } } On Thu, 10 May 2007 10:42:01 -0500 } hyi-at-emory.edu wrote: } } ---------------------------------------------------------------------- } } ------ } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } } America } } To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------- } } ------ } } Dear All: } } I am so glad to see the discussion on the contrast issue } } continues as I have been hearing more and more reports on this } } problem and I myself have been experiencing it too. Many hypotheses } } have been suggested on the causes of the problem but it seems that } } there were always evidences supporting different arguments. } } Practically, besides using short dehydration and infiltration times } } for monolayer cells, I now HAVE TO use potassium ferrocyanide with } } OsO4 in order to get good contrast (even for bulk tissue). When I } } find out the contrast is low after sections are being cut, I } } re-counterstain sections with 15% UA in methanol (it can give a } } muddy appearance if it is over done). In addition, I make sure to } } use a relatively fresh OsO4 stock solution. I have been } } working in EM core facilities for a long time and have dealt with } } all kinds of samples. One question I have for people like me is } } whether this is a new problem or has always been this way. I did not } } remember contrast was so much an issue in the good old days even } } with standard dehydration times and when OsO4 was used alone. } } Personally I think this problem has something to do with the } } reagents we use now days, particularly OsO4. I did notice the results } } were different when I use different OsO4 (liquid vs. crystal, old } } vs. fresh….). I am wondering what other people’s experiences are } } with OsO4 and how OsO4 solution is prepared and stored in different } } laboratories. I would really love to hear from chemist more details } } on how OsO4 works and from vendors if there has been any change in } } their sources for OsO4. } } Thank all very much } } Hong } } Emory SOM EM } } On May 10, 2007, at 8:10 AM, malcolm.haswell-at-sunderland.ac.uk wrote: } } } } } } --------------------------------------------------------------------- } } } -- ----- } } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } } } America } } } To Subscribe/Unsubscribe -- } } } http://www.microscopy.com/MicroscopyListserver } } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } --------------------------------------------------------------------- } } } -- ----- } } } } } } Dear Tom } } } } } } I just thought I should mention something that I think I read years } } } ago } } } in one of M.A. Hayat's books. He mentions that cacodylate and some } } } other } } } buffers when incorporated into a fixative produce higher contrast } } } because they are more extractive whereas phosphate buffers probably } } } preserve cell contents better but at the expense of contrast. } } } } } } I just thought that I should add this because you mention Sorenson's } } } buffer in the fixative. I'm sure that many of the other points have } } } an } } } effect but just wondered if the buffer could be contributing to your } } } problems as well. } } } } } } Malcolm } } } } } } } } } Malcolm Haswell } } } e.m. unit } } } School of Health, Natural and Social Sciences } } } Fleming Building } } } University of Sunderland } } } Tyne & Wear } } } SR1 3SD } } } UK } } } e-mail: malcolm.haswell-at-sunderland.ac.uk } } } } } } } } } ----- Original Message ----- } } } X-from: nizets2-at-yahoo.com } } } Date: Thursday, May 10, 2007 12:23 pm } } } Subject: [Microscopy] Re: Help with enhancing contrast of } } } Mitochondrial } } } cristae } } } } } } } } } } } } } } } } } } } -------------------------------------------------------------------- } } } } -------- } } } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } } } } AmericaTo Subscribe/Unsubscribe -- } } } } http://www.microscopy.com/MicroscopyListserverOn-Line Help } } } } http://www.microscopy.com/MicroscopyListserver/FAQ.html------------- } } } } --------------------------------------------------------------- } } } } } } } } Dear Tom, } } } } } } } } I agree with the other comments: Ferrocyanide, } } } } decreasing dehydration times (penetration is immediate } } } } in monolayers) and using picric acid probably can } } } } improve membrane contrast. } } } } (personally I already tried UAc in methanol without } } } } satisfaction) } } } } } } } } Please share your experience with us. } } } } } } } } Best regards, } } } } } } } } Stephane } } } } } } } } --- tbargar-at-unmc.edu wrote: } } } } } } } } } } } } } } } } } } } } } } } } } } } } -------------------------------------------------------------------- } } } } -------- } } } } } The Microscopy ListServer -- CoSponsor: The } } } } } Microscopy Society of America } } } } } To Subscribe/Unsubscribe -- } } } } } http://www.microscopy.com/MicroscopyListserver } } } } } On-Line Help } } } } } } } } } http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } } } } -------------------------------------------------------------------- } } } } -------- } } } } } } } } } } } } } } } Dear listers, } } } } } } } } } } Anyone out there have any advice on how to enhance } } } } } the contrast and } } } } } definition of the membranes of the cristae of } } } } } mitochondria? The samples } } } } } brought to me are monolayer cell cultures of cancer } } } } } cells grown on } } } } } Thermanox coverslips. This is how I'm currently } } } } } processing the samples: } } } } } Primary fixation is 2% glutaraldehyde, 2% } } } } } paraformaldehyde, 0.5% acrolein } } } } } in 0.1M Sorensen's phosphate buffer pH 7.2. } } } } } Post-fixation in 1%Osmium } } } } } Tetroxide in 0.1M Sorensen's phosphate buffer. } } } } } Dehydration in 50, 70, 90, } } } } } 95, 100%X3 ethanol solutions. embedding in } } } } } Araldite. Sections are stained } } } } } with 2% uranyl acetate aqueous 15 minutes and } } } } } Reynold's lead citrate 10 } } } } } minutes. The density of the cytoplasm and } } } } } mitochondrial matrix are } } } } } similar with the result that the contrast of the } } } } } mitochondria is similar to } } } } } the cytoplasm. The mitochondria and it's membranes } } } } } (outer and that of the } } } } } cristae) don't really "stand out". The researchers } } } } } involved want to see } } } } } really contrasty mitochondrial cristae. } } } } } } } } } } The next thing I'm going to try is post-fixation } } } } } with a mix of osmium } } } } } tetroxide and potassium ferrocyanide. } } } } } } } } } } Anyone have any other ideas? Thanks in advance. } } } } } } } } } } } } } } } Tom Bargar } } } } } University of Nebraska Medical Center } } } } } Core Electron Microscopy Research Facility } } } } } 986395 Nebraska Medical Center } } } } } Omaha, NE 68198-6395 } } } } } 402-559-7347 } } } } } tbargar-at-unmc.edu } } } } } } } } } } } } } } } ==============================Original } } } } } Headers============================== } } } } } 8, 20 -- From tbargar-at-unmc.edu Tue May 1 15:39:39 } } } } } 2007 } } } } } 8, 20 -- Received: from zixvpm02.unmc.edu } } } } } (zixvpm02.unmc.edu [192.198.54.127]) } } } } } 8, 20 -- by ns.microscopy.com } } } } } (8.12.11.20060308/8.12.8) with ESMTP id } } } } } l41KddQH023357 } } } } } 8, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, } } } } } 1 May 2007 15:39:39 -0500 } } } } } 8, 20 -- Received: from zixvpm02.unmc.edu (ZixVPM } } } } } [127.0.0.1]) } } } } } 8, 20 -- by Outbound.unmc.edu (Proprietary) with } } } } } ESMTP id 8A304A0061 } } } } } 8, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, } } } } } 1 May 2007 15:39:39 -0500 (CDT) } } } } } 8, 20 -- Received: from unmcnotes01.unmc.edu } } } } } (host-137-197-103-35.unmc.edu [137.197.103.35]) } } } } } 8, 20 -- by zixvpm02.unmc.edu (Proprietary) with } } } } } ESMTP id 72E1039804A } } } } } 8, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, } } } } } 1 May 2007 15:39:38 -0500 (CDT) } } } } } 8, 20 -- Subject: Help with enhancing contrast of } } } } } Mitochondrial cristae } } } } } 8, 20 -- To: Microscopy-at-MSA.Microscopy.Com } } } } } 8, 20 -- X-Mailer: Lotus Notes Release 7.0.1 January } } } } } 17, 2006 } } } } } 8, 20 -- Message-ID: } } } } } } } } } {OFDC8670F1.FEB83264-ON862572CE.006F5771-862572CE.00717D6E-at-unmc.edu} } } } } } 8, 20 -- From: Tom W Bargar {tbargar-at-unmc.edu} } } } } } 8, 20 -- Date: Tue, 1 May 2007 15:39:27 -0500 } } } } } 8, 20 -- X-MIMETrack: Serialize by Router on } } } } } UNMCNOTES01.UNMC.EDU/Servers/UNEBR at 05/01/2007 } } } } } 03:39:38 } } } } } 8, 20 -- PM } } } } } 8, 20 -- MIME-Version: 1.0 } } } } } 8, 20 -- Content-type: text/plain; charset=US-ASCII } } } } } ==============================End of - } } } } } Headers============================== } } } } } } } } } } } } } } } } } __________________________________________________ } } } } Do You Yahoo!? } } } } Tired of spam? Yahoo! Mail has the best spam protection around } } } } http://mail.yahoo.com } } } } } } } } ==============================Original } } } } Headers==============================9, 21 -- From } } } } nizets2-at-yahoo.com Thu May 10 06:15:21 2007 } } } } 9, 21 -- Received: from web37402.mail.mud.yahoo.com } } } } (web37402.mail.mud.yahoo.com [209.191.91.134]) } } } } 9, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP } } } } id l4ABFKJw009245 } } } } 9, 21 -- for {microscopy-at-microscopy.com} ; Thu, 10 May 2007 } } } } 06:15:21 -0500 } } } } 9, 21 -- Received: (qmail 27735 invoked by uid 60001); 10 May 2007 } } } } 11:15:20 -0000 } } } } 9, 21 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; } } } } 9, 21 -- s=s1024; d=yahoo.com; } } } } 9, 21 -- h=X-YMail-OSG:Received:Date:From:Subject:To:Cc:In-Reply- } } } } To:MIME-Version:Content-Type:Content-Transfer-Encoding:Message-ID; } } } } 9, 21 -- } } } } } } } b=xdVFiLCF4Y+sO8r3CpubJCf2uWbS/tsa8x0ab/ } } } hPl+5HFUOENjJ6yIvxMiFISE4+rsVrEo+9g9amUFa4qIQOAGV0FBzymuaE0sNVDp0SglZ } } } lj } } } Ly8sAz3+WIuaj+c/D2DZnTf4n+Vivh1RJ9XqSQvZy3HvK1FSpB028byJ8YI300=;9, } } } 21 -- X-YMail-OSG: } } } mktHeDIVM1kftlmU8fPc3P3OX17YmyvCJT352ac98PdSr0qhC7X.fCqkPbxI6mkxa09Gc } } } WQ } } } MmEhQCWAHO8TzXOi4W.ueJ.8eBX54nP.Y7I3f9c1fcF6eyrST0RA3CoDbTEtqWbHpql.V } } } e3 3A65i7tH.ZVhYJ5Jn8TfeDlO7pDP4W.DeEznuXRaE- } } } } 9, 21 -- Received: from [80.122.101.102] by } } } } web37402.mail.mud.yahoo.com via HTTP; Thu, 10 May 2007 04:15:20 PDT } } } } 9, 21 -- Date: Thu, 10 May 2007 04:15:20 -0700 (PDT) } } } } 9, 21 -- From: Stephane Nizet {nizets2-at-yahoo.com} } } } } 9, 21 -- Subject: Re: [Microscopy] Help with enhancing contrast of } } } } Mitochondrial cristae } } } } 9, 21 -- To: tbargar-at-unmc.edu } } } } 9, 21 -- Cc: microscopy-at-microscopy.com } } } } 9, 21 -- In-Reply-To: } } } } {200705012045.l41Kjoma031136-at-ns.microscopy.com} } } } } 9, 21 -- MIME-Version: 1.0 } } } } 9, 21 -- Content-Type: text/plain; charset=iso-8859-1 } } } } 9, 21 -- Content-Transfer-Encoding: 8bit } } } } 9, 21 -- Message-ID: {635982.26761.qm-at-web37402.mail.mud.yahoo.com} } } } } ==============================End of - } } } } Headers============================== } } } } } } ==============================Original } } } Headers============================== } } } 8, 37 -- From malcolm.haswell-at-sunderland.ac.uk Thu May 10 07:06:52 } } } 2007 } } } 8, 37 -- Received: from max1.sunderland.ac.uk (max1.sunderland.ac.uk } } } [157.228.29.83]) } } } 8, 37 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP } } } id l4AC6os7021493 } } } 8, 37 -- for {Microscopy-at-microscopy.com} ; 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charset=WINDOWS-1252; delsp=yes; } } format=flowed } } 9, 27 -- Message-Id: {8c61afeb0634c079641ccf803afaa602-at-emory.edu} } } 9, 27 -- From: Hong Yi {hyi-at-emory.edu} } } 9, 27 -- Subject: [Microscopy] Help with enhancing contrast of } } Mitochondrial } } 9, 27 -- Date: Thu, 10 May 2007 12:40:07 -0400 } } 9, 27 -- To: microscopy-at-microscopy.com } } 9, 27 -- X-Mailer: Apple Mail (2.622) } } 9, 27 -- X-Brightmail-Tracker: AAAAAA== } } 9, 27 -- Content-Transfer-Encoding: 8bit } } 9, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } } ns.microscopy.com id l4AFelNb029509 } } ==============================End of - } } Headers============================== } } *************************** } Tamara Howard } Cell Biology & Physiology } UNM-HSC } Albuquerque, NM } ***************************
==============================Original Headers============================== 7, 28 -- From hyi-at-emory.edu Thu May 10 11:32:48 2007 7, 28 -- Received: from virginia.cc.emory.edu (virginia.cc.emory.edu [170.140.8.222]) 7, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4AGWmdl032561 7, 28 -- for {Microscopy-at-microscopy.com} ; Thu, 10 May 2007 11:32:48 -0500 7, 28 -- Received: from virginia (virginia [170.140.8.222]) 7, 28 -- by virginia.cc.emory.edu (8.13.8/8.13.8) with ESMTP id l4AGWmRp017410; 7, 28 -- Thu, 10 May 2007 12:32:48 -0400 (EDT) 7, 28 -- Received: from virginia (unknown [127.0.0.1]) 7, 28 -- by virginia (Symantec Mail Security) with ESMTP id 12CF124FC; 7, 28 -- Thu, 10 May 2007 12:32:48 -0400 (EDT) 7, 28 -- X-AuditID: aa8c08de-00000005000004ef-37-4643492e53bb 7, 28 -- Received: from [170.140.233.63] (unknown [170.140.233.63]) 7, 28 -- by virginia (Symantec Mail Security) with ESMTP id E77FC24EA; 7, 28 -- Thu, 10 May 2007 12:32:46 -0400 (EDT) 7, 28 -- In-Reply-To: {web-15022351-at-theta.unm.edu} 7, 28 -- References: {200705101542.l4AFg1eh031668-at-ns.microscopy.com} {web-15022351-at-theta.unm.edu} 7, 28 -- Mime-Version: 1.0 (Apple Message framework v622) 7, 28 -- Content-Type: text/plain; charset=WINDOWS-1252; delsp=yes; format=flowed 7, 28 -- Message-Id: {f7f922faf3a50f16bff1f9b83c83c1ae-at-emory.edu} 7, 28 -- Cc: Tamara A Howard {thoward-at-unm.edu} 7, 28 -- From: Hong Yi {hyi-at-emory.edu} 7, 28 -- Subject: Re: [Microscopy] Help with enhancing contrast of Mitochondrial 7, 28 -- Date: Thu, 10 May 2007 13:32:07 -0400 7, 28 -- To: Microscopy-at-microscopy.com 7, 28 -- X-Mailer: Apple Mail (2.622) 7, 28 -- X-Brightmail-Tracker: AAAAAA== 7, 28 -- Content-Transfer-Encoding: 8bit 7, 28 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4AGWmdl032561 ==============================End of - Headers==============================
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Dr. Hong- I only saw the problem of the membrane "disappearing" in 1978, when I started preparing layers of epithelial cells. I found a way to work around it and haven't (fortunately) rediscovered the problem since then. Carol Heckman Department of Biological Sciences Bowling Green State University
==============================Original Headers============================== 3, 15 -- From heckman-at-bgnet.bgsu.edu Thu May 10 20:53:27 2007 3, 15 -- Received: from smtp01.bgsu.edu (smtp01.bgsu.edu [129.1.5.17]) 3, 15 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4B1rRJP001436 3, 15 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 10 May 2007 20:53:27 -0500 3, 15 -- Received: from localhost (webmail.bgsu.edu [129.1.5.22]) 3, 15 -- by smtp01.bgsu.edu (Switch-3.2.5/Switch-3.1.6) with ESMTP id l4B1rMas006953; 3, 15 -- Thu, 10 May 2007 21:53:22 -0400 (EDT) 3, 15 -- MIME-Version: 1.0 3, 15 -- Date: Thu, 10 May 2007 21:53:22 -0400 3, 15 -- From: "heckman-at-bgnet.bgsu.edu" {heckman-at-bgnet.bgsu.edu} 3, 15 -- Message-Id: {1178848402-26100.00024.00758-smmsdV2.1.6-at-smtp.bgsu.edu} 3, 15 -- X-SMMS-Source: 72.241.89.214 3, 15 -- To: Microscopy-at-MSA.Microscopy.Com, {hyi-at-emory.edu} 3, 15 -- Subject: Re: [Microscopy] Help with enhancing contrast of Mitochondrial 3, 15 -- In-Reply-To: {200705101541.l4AFfYqc030814-at-ns.microscopy.com} ==============================End of - Headers==============================
I'll try to reply to both your questions with this one post.
Because I use and maintain PIPS does not make me an "expert" at ion milling. I have noticed a few things that do/do not happen under the different milling conditions we work with.
Low kv milling seems to give lower instances of twinning and stacking faults in the thin films I have worked with. But I do not think that milling damage goes away at lower kVs. I see a background of "mottling" in the films I have imaged. This is what I assume is the damage from milling and occurs at 5kV milling as well as 2.6kV milling.
The milling times are so long for the reason that most of the thin-films-on-glass (silicon) are created to study the behavior as deposited - no anneal. This makes the films fragile to the point that even the technique of encasing the sample in a brass tube does not work - the glue bond bonding the coupons together is greater than the bonding of the film to the substrate.
For mechanically prepared films (which our lead microscopist prefers over FIB cut samples) I had to come up with a way of bonding a glass cover slip to the substrate and trapping the film between. As has been mentioned here, glass behaves poorly in an ion beam. As it becomes very thin, it melts and (usually) pulls the epoxy and film off of the substrate.
The only way to stop the melting is to go to low kVs and a low duty cycle. For most samples this is 3.5kV and 1 minute of milling for every 5 minutes. And as I am only around for 8 hours a day, this makes for a 3 day milling session.
I have been able to prepare sections of Pb (actually PbZrTiOx) that give spectacular lattice (although disappointingly similar to silicon). As far as reducing the time for milling silicon samples, at ten microns thickness and 3kV milling on non-temperature sensitive samples, milling times are usually in the 15 to 20 minute range.
Neal
On Thu, 2007-05-10 at 09:10 -0500, richard.beanland-at-bookham.com wrote: } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Neal, } I wouldn't expect a 10um Si sample to take more than 30-60 mins } to thin in a PIPS. } I can only think you are working at a relatively high milling angle } (} 5 degrees) and so have to turn the beam energy down very low so you } don't damage your sample. Why not experiment with one sample using full } power (6kV) and 3 degrees incidence angle, finishing off with 10 mins at } 2kV? It could increase your throughput more than 20 times! } } Richard } } } } ________________________________________ } Richard Beanland } Materials Analysis } Bookham } Caswell } Towcester } Northants } NN12 8EQ } United Kingdom } Tel. +44 1327 356362 } Fax. +44 1327 356775 } http://www.bookham.com } ________________________________________ } } } -----Original Message----- } X-from: nealzimm-at-cpinternet.com [mailto:nealzimm-at-cpinternet.com] } Sent: 10 May 2007 14:10 } To: Richard Beanland } Subject: [Microscopy] Re: Looking for recommendations about dual beam } ion } } } } } ------------------------------------------------------------------------ } ---- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ } ---- } } John } } I work with four Gatan PIPS daily and have found them to be quite } acceptable in ease of both maintenance and use. } } We currently do maintenance only when the ion guns stop functioning, } at that point the machine is taken out of service and a full cleaning } performed. This happens about every five to six months and leaves } the PIPS out of service for two days (One day to clean, pump down } over night, and alignment the next day). The diaphragm pump needs } new diaphragms once a year (about an hours work) but not too much else. } } Work load is typically one to two ceramic samples per day, with an } occasional silicon or glass sample thrown in. The ceramic samples } are lapped to about six microns and mill for an hour to two hours. } Silicon and glass are left about ten microns thick and take much } longer (up to three days), not so much due to the thickness as to } the low kVs used and letting the sample "cool". } } Gatan offers a digital camera/zoom lens set and an external diaphragm } pump as extras. In my opinion, they are a "must have". } } If you have any further questions please feel free to contact me. } } Neal R. Zimmermann } } } } On Wed, 2007-05-09 at 15:43 -0500, jpchandl-at-mines.edu wrote: } } } } } } } ------------------------------------------------------------------------ } ---- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } ------------------------------------------------------------------------ } ---- } } } } I need to tap in to the wisdom of the list about dual beam argon ion } mills. } } } } We have a Bal-Tec RES100 ion mill that we are having difficulty } maintaining. } } It had been in disuse for a couple of years, and has been repaired and } } upgraded recently, at the factory, to be as close to a RES101 as } possible. } } We are having issues such as the guns becoming contaminated and } needing } } service much more frequently than we think they should. Routine } maintenance } } seems to be difficult, too. We think we are looking at a high } maintenance } } instrument here. I would like to know what kind of experience other } labs } } have had with this instrument. } } } } I would also like to hear from people who have experience } using/maintaining } } the RES100/101 or the Gatan PIPS, or both. We need some comparative } } information, so we can make a decision about how to proceed with our } ion } } milling needs. } } } } Any input would be appreciated. } } } } --John } } } } John Chandler } } Manager, EM Lab } } Colorado School of Mines } } Golden, CO 80401 } } jpchandl-at-mines.edu } } 303-384-2203 } } } } } } } } } } ==============================Original } Headers============================== } } 9, 19 -- From jpchandl-at-mines.edu Wed May 9 15:42:15 2007 } } 9, 19 -- Received: from inspire.Mines.EDU (inspire.Mines.EDU } [138.67.130.5]) } } 9, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id l49KgEDO003975 } } 9, 19 -- for {microscopy-at-microscopy.com} ; Wed, 9 May 2007 } 15:42:14 -0500 } } 9, 19 -- Received: from emlab1 (emlab1.Mines.EDU [138.67.28.173]) } } 9, 19 -- by inspire.Mines.EDU (8.13.1/8.13.1) with ESMTP id } l49KgD7T016786 } } 9, 19 -- for {microscopy-at-microscopy.com} ; Wed, 9 May 2007 } 14:42:13 -0600 } } 9, 19 -- From: "John Chandler" {jpchandl-at-mines.edu} } } 9, 19 -- To: {microscopy-at-microscopy.com} } } 9, 19 -- Subject: Looking for recommendations about dual beam ion } mills } } 9, 19 -- Date: Wed, 9 May 2007 14:42:13 -0600 } } 9, 19 -- Message-ID: {001e01c7927a$861c8570$ad1c438a-at-mines.edu} } } 9, 19 -- MIME-Version: 1.0 } } 9, 19 -- Content-Type: text/plain; } } 9, 19 -- charset="us-ascii" } } 9, 19 -- Content-Transfer-Encoding: 7bit } } 9, 19 -- X-Mailer: Microsoft Office Outlook 11 } } 9, 19 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 } } 9, 19 -- Thread-Index: AceSeoX7iyPEBSG3T3O4yAVZe0waUw== } } ==============================End of - } Headers============================== } } } ==============================Original } Headers============================== } 11, 24 -- From nealzimm-at-cpinternet.com Thu May 10 08:08:28 2007 } 11, 24 -- Received: from cpinternet.com (mail.cpinternet.com } [209.240.224.81]) } 11, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id l4AD8S9W001596 } 11, 24 -- for {microscopy-at-microscopy.com} ; Thu, 10 May 2007 } 08:08:28 -0500 } 11, 24 -- Received: from [192.168.1.102] (unverified [216.251.191.121]) } 11, 24 -- by cpinternet.com (SurgeMail 3.8j) with ESMTP id } 171217060-1881964 } 11, 24 -- for multiple; Thu, 10 May 2007 08:08:27 -0500 CDT } 11, 24 -- Subject: Re: [Microscopy] Looking for recommendations about } dual beam ion } 11, 24 -- mills } 11, 24 -- From: Neal Zimmermann {nealzimm-at-cpinternet.com} } 11, 24 -- To: jpchandl-at-mines.edu, microscopy-at-microscopy.com } 11, 24 -- In-Reply-To: {200705092043.l49Kh2W8005876-at-ns.microscopy.com} } 11, 24 -- References: {200705092043.l49Kh2W8005876-at-ns.microscopy.com} } 11, 24 -- Content-Type: text/plain } 11, 24 -- Date: Thu, 10 May 2007 08:08:14 -0500 } 11, 24 -- Message-Id: {1178802494.3663.19.camel-at-localhost.localdomain} } 11, 24 -- Mime-Version: 1.0 } 11, 24 -- X-Mailer: Evolution 2.8.3 (2.8.3-2.fc6) } 11, 24 -- Content-Transfer-Encoding: 7bit } 11, 24 -- X-Authenticated-User: nealzimm-at-cpinternet.com } 11, 24 -- X-NotAscii: charset=us-ascii } 11, 24 -- X-Encryption: SSL encrypted } 11, 24 -- X-IP-stats: No info recorded yet ip=216.251.191.121 } 11, 24 -- X-Originating-IP: 216.251.191.121 } ==============================End of - } Headers============================== } } ======================================================================= } This e-mail is intended for the person it is addressed to only. 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Any use, } forwarding, printing or copying of this message is strictly prohibited. } No part of this message can be considered a request for goods or } services. } ======================================================================= } } } ==============================Original Headers============================== } 23, 34 -- From richard.beanland-at-bookham.com Thu May 10 09:09:28 2007 } 23, 34 -- Received: from mail72.messagelabs.com (mail72.messagelabs.com [193.109.255.147]) } 23, 34 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l4AE9Riq025754 } 23, 34 -- for {microscopy-at-microscopy.com} ; Thu, 10 May 2007 09:09:27 -0500 } 23, 34 -- X-VirusChecked: Checked } 23, 34 -- X-Env-Sender: richard.beanland-at-bookham.com } 23, 34 -- X-Msg-Ref: server-10.tower-72.messagelabs.com!1178806165!33452822!1 } 23, 34 -- X-StarScan-Version: 5.5.10.7.1; banners=bookham.com,-,- } 23, 34 -- X-Originating-IP: [213.249.209.179] } 23, 34 -- Received: (qmail 31853 invoked from network); 10 May 2007 14:09:26 -0000 } 23, 34 -- Received: from unknown (HELO cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI) (213.249.209.179) } 23, 34 -- by server-10.tower-72.messagelabs.com with SMTP; 10 May 2007 14:09:26 -0000 } 23, 34 -- Received: from cas-smx-02.BOOKHAM.ENTERPRISE.PRI ([10.4.0.223]) by cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI with Microsoft SMTPSVC(6.0.3790.1830); } 23, 34 -- Thu, 10 May 2007 15:10:39 +0100 } 23, 34 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 } 23, 34 -- Content-class: urn:content-classes:message } 23, 34 -- MIME-Version: 1.0 } 23, 34 -- Content-Type: text/plain; } 23, 34 -- charset="us-ascii" } 23, 34 -- Subject: RE: [Microscopy] Re: Looking for recommendations about dual beam ion } 23, 34 -- Date: Thu, 10 May 2007 15:10:38 +0100 } 23, 34 -- Message-ID: {9645D3E33E4C6548B12A7B25F611533E46C9B1-at-cas-smx-02.BOOKHAM.ENTERPRISE.PRI} } 23, 34 -- In-Reply-To: {200705101309.l4AD9WRs003477-at-ns.microscopy.com} } 23, 34 -- X-MS-Has-Attach: } 23, 34 -- X-MS-TNEF-Correlator: } 23, 34 -- Thread-Topic: [Microscopy] Re: Looking for recommendations about dual beam ion } 23, 34 -- Thread-Index: AceTBKSYNfWLTtdMSI21epLGVDAwXgABIZoA } 23, 34 -- References: {200705101309.l4AD9WRs003477-at-ns.microscopy.com} } 23, 34 -- From: "Richard Beanland" {richard.beanland-at-bookham.com} } 23, 34 -- To: {nealzimm-at-cpinternet.com} } 23, 34 -- Cc: {microscopy-at-microscopy.com} } 23, 34 -- X-OriginalArrivalTime: 10 May 2007 14:10:39.0130 (UTC) FILETIME=[FCAAEFA0:01C7930C] } 23, 34 -- Content-Transfer-Encoding: 8bit } 23, 34 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4AE9Riq025754 } ==============================End of - Headers==============================
==============================Original Headers============================== 11, 22 -- From nealzimm-at-cpinternet.com Fri May 11 07:47:04 2007 11, 22 -- Received: from cpinternet.com (mail.cpinternet.com [209.240.224.81]) 11, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4BCl3QP022768 11, 22 -- for {microscopy-at-microscopy.com} ; Fri, 11 May 2007 07:47:03 -0500 11, 22 -- Received: from [192.168.1.102] (unverified [216.251.190.54]) 11, 22 -- by cpinternet.com (SurgeMail 3.8j) with ESMTP id 172671156-1881964 11, 22 -- for {microscopy-at-microscopy.com} ; Fri, 11 May 2007 07:47:03 -0500 CDT 11, 22 -- Subject: Re: [Microscopy] Looking for recommendations about dual beam ion 11, 22 -- From: Neal Zimmermann {nealzimm-at-cpinternet.com} 11, 22 -- To: microscopy-at-microscopy.com 11, 22 -- In-Reply-To: {200705101410.l4AEAngB027743-at-ns.microscopy.com} 11, 22 -- References: {200705101410.l4AEAngB027743-at-ns.microscopy.com} 11, 22 -- Content-Type: text/plain 11, 22 -- Date: Fri, 11 May 2007 07:46:51 -0500 11, 22 -- Message-Id: {1178887611.5048.22.camel-at-localhost.localdomain} 11, 22 -- Mime-Version: 1.0 11, 22 -- X-Mailer: Evolution 2.8.3 (2.8.3-2.fc6) 11, 22 -- Content-Transfer-Encoding: 7bit 11, 22 -- X-Authenticated-User: nealzimm-at-cpinternet.com 11, 22 -- X-Encryption: SSL encrypted 11, 22 -- X-IP-stats: Incoming Last 2, First 1, in=4, out=0, spam=0 ip=216.251.190.54 11, 22 -- X-Originating-IP: 216.251.190.54 ==============================End of - Headers==============================
~ 8 mm apart? Did I read this correctly? Not microns? We use a Disco saw routinely. If it's ~ 8 mm apart, you can use any diamond saw. If it's microns you need a FIB.
John Mardinly Intel Corporation This email does not represent an opinion of Intel Corporation.
-----Original Message----- X-from: peter.tomic-at-renwireless.com [mailto:peter.tomic-at-renwireless.com] Sent: Tuesday, May 08, 2007 10:33 AM To: Mardinly, John
Microscopy Folks,
I'd like to get some input on sample preparation with respect to silicon.
PROBLEM:
I have an FEI XL-50 FESEM that was originally designed to accept flat samples, essentially silicon wafers. It does not have what one would consider a typical exchange port. The exchange port is robotically controlled, and the maximum sample height, including stub, must be 5 mm or less. I must view these samples on edge, i.e. 90 degrees. This presents a problem in that I must cleave two parallel sections very close to each other. Perhaps diamond cutters can do this, but my hands are too shaky.
QUESTION:
Is there a device, or method, that would allow me to make these cleaves ~ 8 mm apart with any reasonable control? I found stubs that are low profile, 38 and 90 degrees, from the nice people at Ted Pella, but I still have this issue of doing two parallel cleaves very close together.
Just for info. purposes I am in the silicon MEMS development arena.
If you feel your reply is of general interest to this community, please reply to all, or you may contact me directly.
Regards to all in this small world,
Peter Tomic Renaissance Wireless Corp.
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==============================Original Headers============================== 13, 25 -- From peter.tomic-at-renwireless.com Tue May 8 12:33:06 2007 13, 25 -- Received: from mail.rw.local (4.piok2.xdsl.nauticom.net [209.195.176.101]) 13, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l48HX6om005343 13, 25 -- for {Microscopy-at-microscopy.com} ; Tue, 8 May 2007 12:33:06 -0500 13, 25 -- Received: from localhost (localhost [127.0.0.1]) 13, 25 -- by mail.rw.local (Postfix) with ESMTP id 97112241A0 13, 25 -- for {Microscopy-at-microscopy.com} ; Tue, 8 May 2007 13:33:05 -0400 (EDT) 13, 25 -- Received: from mail.rw.local ([127.0.0.1]) 13, 25 -- by localhost (mail.rw.local [127.0.0.1]) (amavisd-new, port 10024) with ESMTP 13, 25 -- id 30832-08 for {Microscopy-at-microscopy.com} ; 13, 25 -- Tue, 8 May 2007 13:33:05 -0400 (EDT) 13, 25 -- Received: from [127.0.0.1] (unknown [10.11.10.142]) 13, 25 -- by mail.rw.local (Postfix) with ESMTP id 2E11024168 13, 25 -- for {Microscopy-at-microscopy.com} ; Tue, 8 May 2007 13:33:05 -0400 (EDT) 13, 25 -- Message-ID: {4640B45E.6040307-at-renwireless.com} 13, 25 -- Date: Tue, 08 May 2007 13:33:18 -0400 13, 25 -- From: Peter Tomic {peter.tomic-at-renwireless.com} 13, 25 -- Organization: Renaissance Wireless Corp. 13, 25 -- User-Agent: Thunderbird 1.5.0.10 (Windows/20070221) 13, 25 -- MIME-Version: 1.0 13, 25 -- To: Microscopy-at-microscopy.com 13, 25 -- Subject: Silicon Cross-section sample preparation 13, 25 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 13, 25 -- Content-Transfer-Encoding: 7bit 13, 25 -- X-Virus-Scanned: amavisd-new at renwireless.com ==============================End of - Headers==============================
==============================Original Headers============================== 21, 41 -- From john.mardinly-at-intel.com Fri May 11 11:52:10 2007 21, 41 -- Received: from mga03.intel.com (mga03.intel.com [143.182.124.21]) 21, 41 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4BGq9aJ017647 21, 41 -- for {Microscopy-at-msa.microscopy.com} ; Fri, 11 May 2007 11:52:09 -0500 21, 41 -- Received: from azsmga001.ch.intel.com ([10.2.17.19]) 21, 41 -- by azsmga101.ch.intel.com with ESMTP; 11 May 2007 09:52:06 -0700 21, 41 -- X-ExtLoop1: 1 21, 41 -- X-IronPort-AV: E=Sophos;i="4.14,523,1170662400"; 21, 41 -- d="scan'208";a="227063762" 21, 41 -- Received: from fmsmsxpoc001.fm.intel.com (HELO fmsmsxpoc001.amr.corp.intel.com) ([132.233.49.22]) 21, 41 -- by azsmga001.ch.intel.com with ESMTP; 11 May 2007 09:52:06 -0700 21, 41 -- Received: from fmsmsx334.amr.corp.intel.com (132.233.42.1) by 21, 41 -- fmsmsxpoc001.amr.corp.intel.com (132.233.49.22) with Microsoft SMTP Server id 21, 41 -- 8.0.685.24; Fri, 11 May 2007 09:52:03 -0700 21, 41 -- Received: from fmsmsx333.amr.corp.intel.com ([132.233.42.2]) by 21, 41 -- fmsmsx334.amr.corp.intel.com with Microsoft SMTPSVC(6.0.3790.1830); Fri, 11 21, 41 -- May 2007 09:52:03 -0700 21, 41 -- Received: from scsmsx411.amr.corp.intel.com ([10.3.90.30]) by 21, 41 -- fmsmsx333.amr.corp.intel.com with Microsoft SMTPSVC(6.0.3790.1830); Fri, 11 21, 41 -- May 2007 09:52:02 -0700 21, 41 -- Received: from scsmsx415.amr.corp.intel.com ([10.3.90.34]) by 21, 41 -- scsmsx411.amr.corp.intel.com with Microsoft SMTPSVC(6.0.3790.1830); Fri, 11 21, 41 -- May 2007 09:52:02 -0700 21, 41 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 21, 41 -- Content-Class: urn:content-classes:message 21, 41 -- MIME-Version: 1.0 21, 41 -- Content-Type: text/plain; charset="us-ascii" 21, 41 -- Subject: RE: [Microscopy] Silicon Cross-section sample preparation 21, 41 -- Date: Fri, 11 May 2007 09:52:02 -0700 21, 41 -- Message-ID: {F3CB8931ABF8294DB889E977150CAD68705D13-at-scsmsx415.amr.corp.intel.com} 21, 41 -- In-Reply-To: {200705081733.l48HXCwi005463-at-ns.microscopy.com} 21, 41 -- X-MS-Has-Attach: 21, 41 -- X-MS-TNEF-Correlator: 21, 41 -- Thread-Topic: [Microscopy] Silicon Cross-section sample preparation 21, 41 -- thread-index: AceRlvbRaPk7gaA0T/uFk3LNuosddwCVT62g 21, 41 -- From: "Mardinly, John" {john.mardinly-at-intel.com} 21, 41 -- To: {peter.tomic-at-renwireless.com} 21, 41 -- CC: {Microscopy-at-msa.microscopy.com} 21, 41 -- X-OriginalArrivalTime: 11 May 2007 16:52:02.0270 (UTC) FILETIME=[B2AE97E0:01C793EC] 21, 41 -- Content-Transfer-Encoding: 8bit 21, 41 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4BGq9aJ017647 ==============================End of - Headers==============================
Larry- That's the kind of reference that police forensics departments should have. Try contacting libraries at places like John Jay College of Criminal Justice here in NYC (its part of the City University of NY system), or simialr schools near you. Lee
} } Can anyone recommend references on identifying mammalian species by } analysis of their hair? } } } Thanks, } Larry Zagorski
-- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Weill-Cornell Medical College
I have an inquiry for a source for charcoal marker beads of about 30µm in size. These are to be used to mark plant roots to determine growth rate. We have a paper that used this technique but they did not give a source for the charcoal beads.
I have not been able to reach the authors so hoped one of you would have a source.
Debby
Debby Sherman, Director Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www.agriculture.purdue.edu/microscopy
==============================Original Headers============================== 9, 22 -- From dsherman-at-purdue.edu Fri May 11 12:31:34 2007 9, 22 -- Received: from 1061exfe01.adpc.purdue.edu (1061exfe01.adpc.purdue.edu [128.210.63.221]) 9, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4BHVWGq025053 9, 22 -- for {microscopy-at-microscopy.com} ; Fri, 11 May 2007 12:31:33 -0500 9, 22 -- Received: from exchange.purdue.edu ([172.21.6.23]) by 1061exfe01.adpc.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 9, 22 -- Fri, 11 May 2007 13:31:30 -0400 9, 22 -- Received: from 128.210.161.94 ([128.210.161.94]) by EXCH04.purdue.lcl ([172.21.6.26]) via Exchange Front-End Server exch.itap.purdue.edu ([128.210.63.104]) with Microsoft Exchange Server HTTP-DAV ; 9, 22 -- Fri, 11 May 2007 17:31:30 +0000 9, 22 -- User-Agent: Microsoft-Entourage/11.3.3.061214 9, 22 -- Date: Fri, 11 May 2007 13:31:29 -0400 9, 22 -- Subject: Charcoal markers 9, 22 -- From: Debby Sherman {dsherman-at-purdue.edu} 9, 22 -- To: "message to: MSA list" {microscopy-at-microscopy.com} 9, 22 -- Message-ID: {C26A20B1.1C050%dsherman-at-purdue.edu} 9, 22 -- Thread-Topic: Charcoal markers 9, 22 -- Thread-Index: AceT8jVcc/A93P/lEduiwgARJN08Mg== 9, 22 -- Mime-version: 1.0 9, 22 -- Content-type: text/plain; 9, 22 -- charset="ISO-8859-1" 9, 22 -- X-OriginalArrivalTime: 11 May 2007 17:31:30.0712 (UTC) FILETIME=[36622980:01C793F2] 9, 22 -- Content-Transfer-Encoding: 8bit 9, 22 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4BHVWGq025053 ==============================End of - Headers==============================
I'd like to contribute a little to the ion milling damage issue being discussed. I think that I have used every brand of ion mill on the market in my career at some point or another. When I was with PPG, glass substrates were the only cross section samples that I dealt with and they were the most tricky. With those samples, I had the most trouble with the PIPS(TM) instrument. If you look at the milling rates with other instruments, the PIPS(TM) Penning ion guns are similar in ion sputtering rates to other ion guns in other companies' ion mills (except the Technoorg-Linda gun with the Einzel focusing lens). It is actually very hard to compare them because everyone does it differently. I believe that the issue wasn't one of the ion beam, but so for the design of the clamp-type DuoPost(TM) that I was using to mill at low angles. The DuoPost(TM) does not have a large mass, and the gripping method does not have a great thermal path to extract heat away from the sample. Glass also doesn't have a great thermal conductivity. If I did not operate the PIPS with lowered ion gun performance parameters, I would soften my glass samples and they would soften and "droop down" during milling. This did not occur with any other ion mill that I used that had a more massive sample holder with better heat path. These other mills included the ones from BalTec (used extensively), EA Fischione (used extensively), and Technoorg-Linda (only a few samples).
With respect to ion damage from ion milling, I think that Arpad Barna's group's work is definitive on the topic. He ion milled Si and GaAs samples at different energies at 5 degrees and then immediately evaporated Al on top to protect the samples from oxidation when they were taken out of the ion mill. He then prepared cross sections of those samples and measured the thickness of the amorphous damage region on the surface due to the ion milling by using HREM. For Si, they saw that at 3 kV (about the limit for conventional ion guns) an amorphous region 120 Angstroms was formed. Using the Technoorg-Linda low energy gun at 250 eV, they achieved an amorphous region of about 10 Angstroms. (A. Barna, B. Pecz; Amorphisation and surface morphology development at low-energy ion milling; Ultramicroscopy 70, 1998.) In other studies and examples, they have also shown that the mottling from ion milled and FIB samples using the low energy guns can be completely eliminated. I have some of the data from that work if you are interested in seeing the results.
Disclaimer: South Bay Technology, Inc. is the exclusive distributer for Technoorg-Linda products in the United States.
-Scott
Scott D. Walck, Ph.D. Technical Director South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
US Toll Free: 1-800-728-2233 Tel: (949) 492-2600 Fax: (949) 492-1499
www.southbaytech.com walck-at-southbaytech.com
-----Original Message----- X-from: nealzimm-at-cpinternet.com [mailto:nealzimm-at-cpinternet.com] Sent: Friday, May 11, 2007 5:53 AM To: Walck-at-SouthBayTech.com
Richard, Witold;
I'll try to reply to both your questions with this one post.
Because I use and maintain PIPS does not make me an "expert" at ion milling. I have noticed a few things that do/do not happen under the different milling conditions we work with.
Low kv milling seems to give lower instances of twinning and stacking faults in the thin films I have worked with. But I do not think that milling damage goes away at lower kVs. I see a background of "mottling" in the films I have imaged. This is what I assume is the damage from milling and occurs at 5kV milling as well as 2.6kV milling.
The milling times are so long for the reason that most of the thin-films-on-glass (silicon) are created to study the behavior as deposited - no anneal. This makes the films fragile to the point that even the technique of encasing the sample in a brass tube does not work - the glue bond bonding the coupons together is greater than the bonding of the film to the substrate.
For mechanically prepared films (which our lead microscopist prefers over FIB cut samples) I had to come up with a way of bonding a glass cover slip to the substrate and trapping the film between. As has been mentioned here, glass behaves poorly in an ion beam. As it becomes very thin, it melts and (usually) pulls the epoxy and film off of the substrate.
The only way to stop the melting is to go to low kVs and a low duty cycle. For most samples this is 3.5kV and 1 minute of milling for every 5 minutes. And as I am only around for 8 hours a day, this makes for a 3 day milling session.
I have been able to prepare sections of Pb (actually PbZrTiOx) that give spectacular lattice (although disappointingly similar to silicon). As far as reducing the time for milling silicon samples, at ten microns thickness and 3kV milling on non-temperature sensitive samples, milling times are usually in the 15 to 20 minute range.
Neal
On Thu, 2007-05-10 at 09:10 -0500, richard.beanland-at-bookham.com wrote: } } } ---------------------------------------------------------------------- } ------ The Microscopy ListServer -- CoSponsor: The Microscopy Society } of America To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } Neal, } I wouldn't expect a 10um Si sample to take more than 30-60 mins to } thin in a PIPS. } I can only think you are working at a relatively high milling angle } (} 5 degrees) and so have to turn the beam energy down very low so you } don't damage your sample. Why not experiment with one sample using } full power (6kV) and 3 degrees incidence angle, finishing off with 10 } mins at 2kV? It could increase your throughput more than 20 times! } } Richard } } } } ________________________________________ } Richard Beanland } Materials Analysis } Bookham } Caswell } Towcester } Northants } NN12 8EQ } United Kingdom } Tel. +44 1327 356362 } Fax. +44 1327 356775 } http://www.bookham.com } ________________________________________ } } } -----Original Message----- } X-from: nealzimm-at-cpinternet.com [mailto:nealzimm-at-cpinternet.com] } Sent: 10 May 2007 14:10 } To: Richard Beanland } Subject: [Microscopy] Re: Looking for recommendations about dual beam } ion } } } } } ---------------------------------------------------------------------- } -- } ---- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -- } ---- } } John } } I work with four Gatan PIPS daily and have found them to be quite } acceptable in ease of both maintenance and use. } } We currently do maintenance only when the ion guns stop functioning, } at that point the machine is taken out of service and a full cleaning } performed. This happens about every five to six months and leaves the } PIPS out of service for two days (One day to clean, pump down over } night, and alignment the next day). The diaphragm pump needs new } diaphragms once a year (about an hours work) but not too much else. } } Work load is typically one to two ceramic samples per day, with an } occasional silicon or glass sample thrown in. The ceramic samples are } lapped to about six microns and mill for an hour to two hours. } Silicon and glass are left about ten microns thick and take much } longer (up to three days), not so much due to the thickness as to the } low kVs used and letting the sample "cool". } } Gatan offers a digital camera/zoom lens set and an external diaphragm } pump as extras. In my opinion, they are a "must have". } } If you have any further questions please feel free to contact me. } } Neal R. Zimmermann } } } } On Wed, 2007-05-09 at 15:43 -0500, jpchandl-at-mines.edu wrote: } } } } } } } ---------------------------------------------------------------------- } -- } ---- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } ---------------------------------------------------------------------- } -- } ---- } } } } I need to tap in to the wisdom of the list about dual beam argon ion } mills. } } } } We have a Bal-Tec RES100 ion mill that we are having difficulty } maintaining. } } It had been in disuse for a couple of years, and has been repaired } } and upgraded recently, at the factory, to be as close to a RES101 as } possible. } } We are having issues such as the guns becoming contaminated and } needing } } service much more frequently than we think they should. Routine } maintenance } } seems to be difficult, too. We think we are looking at a high } maintenance } } instrument here. I would like to know what kind of experience other } labs } } have had with this instrument. } } } } I would also like to hear from people who have experience } using/maintaining } } the RES100/101 or the Gatan PIPS, or both. We need some comparative } } information, so we can make a decision about how to proceed with our } ion } } milling needs. } } } } Any input would be appreciated. } } } } --John } } } } John Chandler } } Manager, EM Lab } } Colorado School of Mines } } Golden, CO 80401 } } jpchandl-at-mines.edu } } 303-384-2203 } } } } } } } } } } ==============================Original } Headers============================== } } 9, 19 -- From jpchandl-at-mines.edu Wed May 9 15:42:15 2007 9, 19 -- } } Received: from inspire.Mines.EDU (inspire.Mines.EDU } [138.67.130.5]) } } 9, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id l49KgEDO003975 } } 9, 19 -- for {microscopy-at-microscopy.com} ; Wed, 9 May 2007 } 15:42:14 -0500 } } 9, 19 -- Received: from emlab1 (emlab1.Mines.EDU [138.67.28.173]) } } 9, 19 -- by inspire.Mines.EDU (8.13.1/8.13.1) with ESMTP id } l49KgD7T016786 } } 9, 19 -- for {microscopy-at-microscopy.com} ; Wed, 9 May 2007 } 14:42:13 -0600 } } 9, 19 -- From: "John Chandler" {jpchandl-at-mines.edu} 9, 19 -- To: } } {microscopy-at-microscopy.com} 9, 19 -- Subject: Looking for } } recommendations about dual beam ion } mills } } 9, 19 -- Date: Wed, 9 May 2007 14:42:13 -0600 9, 19 -- Message-ID: } } {001e01c7927a$861c8570$ad1c438a-at-mines.edu} } } 9, 19 -- MIME-Version: 1.0 } } 9, 19 -- Content-Type: text/plain; } } 9, 19 -- charset="us-ascii" } } 9, 19 -- Content-Transfer-Encoding: 7bit 9, 19 -- X-Mailer: } } Microsoft Office Outlook 11 9, 19 -- X-MimeOLE: Produced By } } Microsoft MimeOLE V6.00.2900.3028 9, 19 -- Thread-Index: } } AceSeoX7iyPEBSG3T3O4yAVZe0waUw== ==============================End } } of - } Headers============================== } } } ==============================Original } Headers============================== } 11, 24 -- From nealzimm-at-cpinternet.com Thu May 10 08:08:28 2007 11, 24 } -- Received: from cpinternet.com (mail.cpinternet.com } [209.240.224.81]) } 11, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id l4AD8S9W001596 } 11, 24 -- for {microscopy-at-microscopy.com} ; Thu, 10 May 2007 } 08:08:28 -0500 } 11, 24 -- Received: from [192.168.1.102] (unverified [216.251.191.121]) } 11, 24 -- by cpinternet.com (SurgeMail 3.8j) with ESMTP id } 171217060-1881964 } 11, 24 -- for multiple; Thu, 10 May 2007 08:08:27 -0500 CDT } 11, 24 -- Subject: Re: [Microscopy] Looking for recommendations about } dual beam ion } 11, 24 -- mills } 11, 24 -- From: Neal Zimmermann {nealzimm-at-cpinternet.com} 11, 24 -- } To: jpchandl-at-mines.edu, microscopy-at-microscopy.com 11, 24 -- } In-Reply-To: {200705092043.l49Kh2W8005876-at-ns.microscopy.com} } 11, 24 -- References: {200705092043.l49Kh2W8005876-at-ns.microscopy.com} } 11, 24 -- Content-Type: text/plain } 11, 24 -- Date: Thu, 10 May 2007 08:08:14 -0500 11, 24 -- Message-Id: } {1178802494.3663.19.camel-at-localhost.localdomain} } 11, 24 -- Mime-Version: 1.0 } 11, 24 -- X-Mailer: Evolution 2.8.3 (2.8.3-2.fc6) 11, 24 -- } Content-Transfer-Encoding: 7bit 11, 24 -- X-Authenticated-User: } nealzimm-at-cpinternet.com 11, 24 -- X-NotAscii: charset=us-ascii 11, 24 } -- X-Encryption: SSL encrypted 11, 24 -- X-IP-stats: No info recorded } yet ip=216.251.191.121 11, 24 -- X-Originating-IP: 216.251.191.121 } ==============================End of - } Headers============================== } } ====================================================================== } = This e-mail is intended for the person it is addressed to only. 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Any use, } forwarding, printing or copying of this message is strictly prohibited. } No part of this message can be considered a request for goods or } services. } ====================================================================== } = } } } ==============================Original } Headers============================== } 23, 34 -- From richard.beanland-at-bookham.com Thu May 10 09:09:28 2007 } 23, 34 -- Received: from mail72.messagelabs.com (mail72.messagelabs.com [193.109.255.147]) } 23, 34 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l4AE9Riq025754 } 23, 34 -- for {microscopy-at-microscopy.com} ; Thu, 10 May 2007 09:09:27 -0500 } 23, 34 -- X-VirusChecked: Checked } 23, 34 -- X-Env-Sender: richard.beanland-at-bookham.com 23, 34 -- } X-Msg-Ref: server-10.tower-72.messagelabs.com!1178806165!33452822!1 } 23, 34 -- X-StarScan-Version: 5.5.10.7.1; banners=bookham.com,-,- 23, } 34 -- X-Originating-IP: [213.249.209.179] 23, 34 -- Received: (qmail } 31853 invoked from network); 10 May 2007 14:09:26 -0000 23, 34 -- } Received: from unknown (HELO cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI) (213.249.209.179) } 23, 34 -- by server-10.tower-72.messagelabs.com with SMTP; 10 May 2007 14:09:26 -0000 } 23, 34 -- Received: from cas-smx-02.BOOKHAM.ENTERPRISE.PRI ([10.4.0.223]) by cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI with Microsoft SMTPSVC(6.0.3790.1830); } 23, 34 -- Thu, 10 May 2007 15:10:39 +0100 } 23, 34 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 23, 34 -- } Content-class: urn:content-classes:message 23, 34 -- MIME-Version: 1.0 } 23, 34 -- Content-Type: text/plain; } 23, 34 -- charset="us-ascii" } 23, 34 -- Subject: RE: [Microscopy] Re: Looking for recommendations } about dual beam ion 23, 34 -- Date: Thu, 10 May 2007 15:10:38 +0100 } 23, 34 -- Message-ID: } {9645D3E33E4C6548B12A7B25F611533E46C9B1-at-cas-smx-02.BOOKHAM.ENTERPRISE. } PRI} 23, 34 -- In-Reply-To: } {200705101309.l4AD9WRs003477-at-ns.microscopy.com} } 23, 34 -- X-MS-Has-Attach: } 23, 34 -- X-MS-TNEF-Correlator: } 23, 34 -- Thread-Topic: [Microscopy] Re: Looking for recommendations } about dual beam ion 23, 34 -- Thread-Index: } AceTBKSYNfWLTtdMSI21epLGVDAwXgABIZoA } 23, 34 -- References: {200705101309.l4AD9WRs003477-at-ns.microscopy.com} } 23, 34 -- From: "Richard Beanland" {richard.beanland-at-bookham.com} 23, } 34 -- To: {nealzimm-at-cpinternet.com} 23, 34 -- Cc: } {microscopy-at-microscopy.com} 23, 34 -- X-OriginalArrivalTime: 10 May } 2007 14:10:39.0130 (UTC) FILETIME=[FCAAEFA0:01C7930C] 23, 34 -- } Content-Transfer-Encoding: 8bit 23, 34 -- X-MIME-Autoconverted: from } quoted-printable to 8bit by ns.microscopy.com id l4AE9Riq025754 } ==============================End of - } Headers==============================
==============================Original Headers============================== 11, 22 -- From nealzimm-at-cpinternet.com Fri May 11 07:47:04 2007 11, 22 -- Received: from cpinternet.com (mail.cpinternet.com [209.240.224.81]) 11, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4BCl3QP022768 11, 22 -- for {microscopy-at-microscopy.com} ; Fri, 11 May 2007 07:47:03 -0500 11, 22 -- Received: from [192.168.1.102] (unverified [216.251.190.54]) 11, 22 -- by cpinternet.com (SurgeMail 3.8j) with ESMTP id 172671156-1881964 11, 22 -- for {microscopy-at-microscopy.com} ; Fri, 11 May 2007 07:47:03 -0500 CDT 11, 22 -- Subject: Re: [Microscopy] Looking for recommendations about dual beam ion 11, 22 -- From: Neal Zimmermann {nealzimm-at-cpinternet.com} 11, 22 -- To: microscopy-at-microscopy.com 11, 22 -- In-Reply-To: {200705101410.l4AEAngB027743-at-ns.microscopy.com} 11, 22 -- References: {200705101410.l4AEAngB027743-at-ns.microscopy.com} 11, 22 -- Content-Type: text/plain 11, 22 -- Date: Fri, 11 May 2007 07:46:51 -0500 11, 22 -- Message-Id: {1178887611.5048.22.camel-at-localhost.localdomain} 11, 22 -- Mime-Version: 1.0 11, 22 -- X-Mailer: Evolution 2.8.3 (2.8.3-2.fc6) 11, 22 -- Content-Transfer-Encoding: 7bit 11, 22 -- X-Authenticated-User: nealzimm-at-cpinternet.com 11, 22 -- X-Encryption: SSL encrypted 11, 22 -- X-IP-stats: Incoming Last 2, First 1, in=4, out=0, spam=0 ip=216.251.190.54 11, 22 -- X-Originating-IP: 216.251.190.54 ==============================End of - Headers==============================
==============================Original Headers============================== 22, 22 -- From walck-at-southbaytech.com Fri May 11 12:42:35 2007 22, 22 -- Received: from flpi102.sbcis.sbc.com (flpi102.sbcis.sbc.com [207.115.20.71]) 22, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4BHgZrx005622 22, 22 -- for {Microscopy-at-microscopy.com} ; Fri, 11 May 2007 12:42:35 -0500 22, 22 -- X-ORBL: [64.169.217.123] 22, 22 -- Received: from dynamicbl8uno3 (adsl-64-169-217-123.dsl.lsan03.pacbell.net [64.169.217.123]) 22, 22 -- by flpi102.sbcis.sbc.com (8.13.8 out.dk.spool/8.13.8) with ESMTP id l4BHgLVU020749; 22, 22 -- Fri, 11 May 2007 10:42:22 -0700 22, 22 -- From: "Scott Walck" {walck-at-southbaytech.com} 22, 22 -- To: {Microscopy-at-microscopy.com} 22, 22 -- Cc: {nealzimm-at-cpinternet.com} 22, 22 -- Subject: RE: [Microscopy] Re: Looking for recommendations about dual beam ion 22, 22 -- Date: Fri, 11 May 2007 10:42:28 -0700 22, 22 -- Message-ID: {002d01c793f3$bf1292f0$7801a8c0-at-dynamicbl8uno3} 22, 22 -- MIME-Version: 1.0 22, 22 -- Content-Type: text/plain; 22, 22 -- charset="us-ascii" 22, 22 -- Content-Transfer-Encoding: 7bit 22, 22 -- X-Mailer: Microsoft Office Outlook 11 22, 22 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 22, 22 -- In-Reply-To: {200705111252.l4BCqYea028524-at-ns.microscopy.com} 22, 22 -- Thread-Index: AceTyz6GLfgJgdXUQD604ooQaJ5lGQAIEt2g ==============================End of - Headers==============================
Roy, did you try using Indium wire to make a seal? Philips used it to make UHV seals at EM 4x0 TEMs. On the other hand the groove should not be too large. You may get Indium wire up to 1mm diameter (or even more).
Best, Stefan Diller
----- Original Message ----- X-from: {rbeavers-at-mail.smu.edu} To: {stefan.diller-at-t-online.de} Sent: Friday, May 11, 2007 7:51 PM
Roy,
In one of my past lives I used to work with a lot of older vacuum equipment that was difficult to get off the shelf O-rings for. I would make my own O-rings with frequency. I used to get viton O-ring material of varying diameters on a spool (like rope).
I guess you could try a larger diameter pre-made O-ring and cut it down, I can't speak about that specifically.
As far as cutting and gluing. I did a diagonal cut using a very sharp razor blade, cutting both ends at the same time. I would glue it with, straight from the local hardware store, Crazy Glue. Then take fine sandpaper - I think I used 600 grit - (but I honestly don't remember that exact detail) and sanded the joint so it was smooth.
I tried butt joints, but never seemed to get them to work, so ended up always doing a diagonal cut. I'm sure others have had success that way, but I didn't.
Good luck.
dj
On Fri, 11 May 2007, rbeavers-at-mail.smu.edu wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Group, } } Has anyone had any experience making a custom Viton O-ring by splicing a } larger one? } } Have a custom chamber operating at 10e-6 torr range and no standard } O-ring seems to fit properly. } } If you have experience with type of splice (butt or diagonal) or glue } used would love to hear about it. } } Thanks } } Roy Beavers } } Southern Methodist University } Department of Geological Sciences } P.O. Box 750395 } Dallas, TX 75275 } Voice: 214-768-2756 } Fax: 214-768-2701 } Email: rbeavers-at-smu.edu } } } ==============================Original Headers============================== } 8, 23 -- From rbeavers-at-mail.smu.edu Fri May 11 12:46:53 2007 } 8, 23 -- Received: from s31xub.systems.smu.edu (s31xub.systems.smu.edu [129.119.70.73]) } 8, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4BHkq8N012896 } 8, 23 -- for {Microscopy-at-microscopy.com} ; Fri, 11 May 2007 12:46:53 -0500 } 8, 23 -- Received: from s31xe7.systems.smu.edu ([129.119.70.140]) by s31xub.systems.smu.edu with Microsoft SMTPSVC(6.0.3790.3959); } 8, 23 -- Fri, 11 May 2007 12:47:16 -0500 } 8, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 } 8, 23 -- Content-class: urn:content-classes:message } 8, 23 -- MIME-Version: 1.0 } 8, 23 -- Content-Type: text/plain; } 8, 23 -- charset="us-ascii" } 8, 23 -- Subject: O-ring help } 8, 23 -- Date: Fri, 11 May 2007 12:47:16 -0500 } 8, 23 -- Message-ID: {F8AF6BF6CD1CC040AF35B0C2D1680BBB021A6F74-at-s31xe7.systems.smu.edu} } 8, 23 -- X-MS-Has-Attach: } 8, 23 -- X-MS-TNEF-Correlator: } 8, 23 -- Thread-Topic: O-ring help } 8, 23 -- Thread-Index: AceT9GoWreqLTSl6SqKSSWHaSbozRg== } 8, 23 -- From: "Beavers, Roy" {rbeavers-at-mail.smu.edu} } 8, 23 -- To: "Microscopy Listserver" {Microscopy-at-microscopy.com} } 8, 23 -- X-OriginalArrivalTime: 11 May 2007 17:47:17.0174 (UTC) FILETIME=[6A84CD60:01C793F4] } 8, 23 -- Content-Transfer-Encoding: 8bit } 8, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4BHkq8N012896 } ==============================End of - Headers============================== }
==============================Original Headers============================== 10, 19 -- From dljones-at-bestweb.net Fri May 11 13:13:55 2007 10, 19 -- Received: from vms040pub.verizon.net (vms040pub.verizon.net [206.46.252.40]) 10, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4BIDsKt008255 10, 19 -- for {Microscopy-at-microscopy.com} ; Fri, 11 May 2007 13:13:55 -0500 10, 19 -- Received: from localhost ([71.247.123.208]) 10, 19 -- by vms040.mailsrvcs.net (Sun Java System Messaging Server 6.2-6.01 (built Apr 10, 19 -- 3 2006)) with ESMTPA id {0JHW002YP2M87KIC-at-vms040.mailsrvcs.net} for 10, 19 -- Microscopy-at-microscopy.com; Fri, 11 May 2007 13:13:25 -0500 (CDT) 10, 19 -- Date: Fri, 11 May 2007 14:14:11 -0400 (Eastern Daylight Time) 10, 19 -- From: "David L. Jones" {dljones-at-bestweb.net} 10, 19 -- Subject: Re: [Microscopy] O-ring help 10, 19 -- In-reply-to: {200705111751.l4BHp2gJ023535-at-ns.microscopy.com} 10, 19 -- X-X-Sender: dljones-at-pop-croton.bestweb.net 10, 19 -- To: rbeavers-at-mail.smu.edu 10, 19 -- Cc: Microscopy-at-microscopy.com 10, 19 -- Message-id: {Pine.WNT.4.64.0705111359450.3084-at-H-F1} 10, 19 -- MIME-version: 1.0 10, 19 -- Content-type: TEXT/PLAIN; charset=US-ASCII; format=flowed 10, 19 -- References: {200705111751.l4BHp2gJ023535-at-ns.microscopy.com} ==============================End of - Headers==============================
Hi Larry You should check ou the FBI website Forensic Science Communications Volume 6 (2004) Microscopy of Hair Part 1: A Practical Guide and Manual for Human Hairs and Microscopy of Hair Part II: A Practical Guide and Manual for Animal Hairs by Douglas W. Deedrick and Sandra L. Koch
We use these as a principle resource in our Imaging and Analysis Course
Rick,
Richard Harris Manager - Imaging, Information and Data Systems Biotron Experimental Climate Change Research Facility University of Western Ontario, London Ontario, CANADA. N6A 5B7 Ph. 519-661-2111 ext. 86780 Fax 519-661-3935 e-mail rjharris-at-uwo.ca web: www.biotron.uwo.ca
-----Original Message----- X-from: LarryZ-at-fai.us [mailto:LarryZ-at-fai.us] Sent: Friday, May 11, 2007 1:20 PM To: rjharris-at-uwo.ca
Can anyone recommend references on identifying mammalian species by analysis of their hair?
Thanks, Larry Zagorski
==============================Original Headers============================== 3, 20 -- From LarryZ-at-fai.us Fri May 11 12:15:37 2007 3, 20 -- Received: from mx.cbeyond.com (smtp.atl.cbeyond.com [66.180.96.29]) 3, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4BHFajE032360 3, 20 -- for {microscopy-at-microscopy.com} ; Fri, 11 May 2007 12:15:37 -0500 3, 20 -- Received: from [69.15.9.254] (port=1513 helo=[192.168.2.5]) 3, 20 -- by mx.cbeyond.com with esmtpa (Exim 4.62) 3, 20 -- (envelope-from {LarryZ-at-fai.us} ) 3, 20 -- id 1HmYja-0000RW-Dq 3, 20 -- for microscopy-at-microscopy.com; Fri, 11 May 2007 13:16:50 -0400 3, 20 -- Message-ID: {4644A4C2.3080802-at-fai.us} 3, 20 -- Date: Fri, 11 May 2007 13:15:46 -0400 3, 20 -- From: Larry Zagorski {LarryZ-at-fai.us} 3, 20 -- Reply-To: LarryZ-at-fai.us 3, 20 -- Organization: FAI 3, 20 -- User-Agent: Thunderbird 1.5.0.10 (Windows/20070221) 3, 20 -- MIME-Version: 1.0 3, 20 -- To: microscopy-at-microscopy.com 3, 20 -- Subject: Mammalian Identified by Hair Imaging 3, 20 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 3, 20 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
==============================Original Headers============================== 15, 26 -- From rjharris-at-uwo.ca Fri May 11 13:42:38 2007 15, 26 -- Received: from uwo.ca (v320-147-lb.its.uwo.ca [129.100.74.147]) 15, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4BIgc6q020178 15, 26 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 11 May 2007 13:42:38 -0500 15, 26 -- Received: from groucho.mail.uwo.pri (brutus.mail.uwo.pri [172.29.32.39]) 15, 26 -- by groucho.mail.uwo.pri 15, 26 -- (Sun Java System Messaging Server 6.2-6.03 (built May 18 2006)) 15, 26 -- with ESMTP id {0JHW00M7A3Z0ICF0-at-groucho.mail.uwo.pri} for 15, 26 -- Microscopy-at-MSA.Microscopy.Com; Fri, 11 May 2007 14:42:36 -0400 (EDT) 15, 26 -- Received: from RICKNOTEBOOK ([129.100.68.106]) 15, 26 -- by groucho.mail.uwo.pri (Sun Java System Messaging Server 6.2-6.03 (built May 15, 26 -- 18 2006)) with ESMTPS id {0JHW00M5E3YZIY30-at-groucho.mail.uwo.pri} for 15, 26 -- Microscopy-at-MSA.Microscopy.Com; Fri, 11 May 2007 14:42:35 -0400 (EDT) 15, 26 -- Date: Fri, 11 May 2007 14:42:35 -0400 15, 26 -- From: Richard Harris {rjharris-at-uwo.ca} 15, 26 -- Subject: RE: [Microscopy] Mammalian Identified by Hair Imaging 15, 26 -- In-reply-to: {200705111719.l4BHJvmG009681-at-ns.microscopy.com} 15, 26 -- To: LarryZ-at-fai.us 15, 26 -- Cc: MSA Listserver {Microscopy-at-MSA.Microscopy.Com} 15, 26 -- Message-id: {0JHW00M5F3YZIY30-at-groucho.mail.uwo.pri} 15, 26 -- MIME-version: 1.0 15, 26 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 15, 26 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 15, 26 -- Content-type: text/plain; charset=us-ascii 15, 26 -- Content-transfer-encoding: 7BIT 15, 26 -- Thread-index: AceT8Jl+IbnfTbYcRJu0SQv3c1OZRgACllcA ==============================End of - Headers==============================
MarcoRubber has a full selection of o-rings. They are online and prompt. This is their URL: http://www.marcorubber.com/ For most rings, I find the cost for 10 to be the same as the cost for one.
There are good charts for ISO, DIN, Italian, British, Japanese, French, Swedish, USA, and standard-metric at this URL: http://mdmetric.com/or/gb01.htm Maryland Metric
regards,
Jim
PS: No commercial connection, just a HAPPY enduser.
PPS: Can't wait for the slew of "O-o-O" messages.
} From mail-at-ns.microscopy.com Fri May 11 13:47:10 2007 } Date: Fri, 11 May 2007 12:47:38 -0500 } To: jquinn-at-www.matscieng.sunysb.edu } From: rbeavers-at-mail.smu.edu } Reply-to: rbeavers-at-mail.smu.edu } X-Resent-From: "Microscopy Listserver" {microscopy-at-microscopy.com} } Subject: [Microscopy] O-ring help } Errors-To: MicroscopyListSpamFilter-at-microscopy.com } X-lewp: MicroscopyListSpam NAGS } } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Group, } } Has anyone had any experience making a custom Viton O-ring by splicing a } larger one? } } Have a custom chamber operating at 10e-6 torr range and no standard } O-ring seems to fit properly. } } If you have experience with type of splice (butt or diagonal) or glue } used would love to hear about it. } } Thanks } } Roy Beavers } } Southern Methodist University } Department of Geological Sciences } P.O. Box 750395 } Dallas, TX 75275 } Voice: 214-768-2756 } Fax: 214-768-2701 } Email: rbeavers-at-smu.edu } } } ==============================Original Headers============================== } 8, 23 -- From rbeavers-at-mail.smu.edu Fri May 11 12:46:53 2007 } 8, 23 -- Received: from s31xub.systems.smu.edu (s31xub.systems.smu.edu [129.119.70.73]) } 8, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4BHkq8N012896 } 8, 23 -- for {Microscopy-at-microscopy.com} ; Fri, 11 May 2007 12:46:53 -0500 } 8, 23 -- Received: from s31xe7.systems.smu.edu ([129.119.70.140]) by s31xub.systems.smu.edu with Microsoft SMTPSVC(6.0.3790.3959); } 8, 23 -- Fri, 11 May 2007 12:47:16 -0500 } 8, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 } 8, 23 -- Content-class: urn:content-classes:message } 8, 23 -- MIME-Version: 1.0 } 8, 23 -- Content-Type: text/plain; } 8, 23 -- charset="us-ascii" } 8, 23 -- Subject: O-ring help } 8, 23 -- Date: Fri, 11 May 2007 12:47:16 -0500 } 8, 23 -- Message-ID: {F8AF6BF6CD1CC040AF35B0C2D1680BBB021A6F74-at-s31xe7.systems.smu.edu} } 8, 23 -- X-MS-Has-Attach: } 8, 23 -- X-MS-TNEF-Correlator: } 8, 23 -- Thread-Topic: O-ring help } 8, 23 -- Thread-Index: AceT9GoWreqLTSl6SqKSSWHaSbozRg== } 8, 23 -- From: "Beavers, Roy" {rbeavers-at-mail.smu.edu} } 8, 23 -- To: "Microscopy Listserver" {Microscopy-at-microscopy.com} } 8, 23 -- X-OriginalArrivalTime: 11 May 2007 17:47:17.0174 (UTC) FILETIME=[6A84CD60:01C793F4] } 8, 23 -- Content-Transfer-Encoding: 8bit } 8, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4BHkq8N012896 } ==============================End of - Headers============================== }
==============================Original Headers============================== 10, 12 -- From jquinn-at-www.matscieng.sunysb.edu Fri May 11 15:51:54 2007 10, 12 -- Received: from www.matscieng.sunysb.edu (www.matscieng.sunysb.edu [129.49.36.33]) 10, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4BKpraB001379 10, 12 -- for {microscopy-at-microscopy.com} ; Fri, 11 May 2007 15:51:54 -0500 10, 12 -- Received: (from jquinn-at-localhost) 10, 12 -- by www.matscieng.sunysb.edu (8.11.6/8.11.6) id l4BKpJB18297 10, 12 -- for microscopy-at-microscopy.com; Fri, 11 May 2007 16:51:19 -0400 10, 12 -- Date: Fri, 11 May 2007 16:51:19 -0400 10, 12 -- From: Jim Quinn {jquinn-at-www.matscieng.sunysb.edu} 10, 12 -- Message-Id: {200705112051.l4BKpJB18297-at-www.matscieng.sunysb.edu} 10, 12 -- To: microscopy-at-microscopy.com 10, 12 -- Subject: re: O-ring help ==============================End of - Headers==============================
Dear Listers: We have a malfunctioning focusing screen on a 100CX II TEM and we have been unable to find the cause of the problem, in spite of phone help from our local JEOL service office and on-site help from an independent service company. Apparently, the motor is receiving about 14 V when it should be receiving 24 V. The screen doesn't lift now, because it has fallen off its track; the motor was constantly running while it was receiving the incorrect voltage. Has anyone seen this problem before, or has a suggestion on how it might be corrected?
Yolande Berta Georgia Tech Center for Nanostructure Characterization and Fabricaton Atlanta, GA 30332-0245
--
==============================Original Headers============================== 5, 30 -- From yb4-at-mail.gatech.edu Fri May 11 22:20:35 2007 5, 30 -- Received: from deliverator8.gatech.edu (deliverator8.gatech.edu [130.207.165.183]) 5, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4C3KZjY018158 5, 30 -- for {microscopy-at-microscopy.com} ; Fri, 11 May 2007 22:20:35 -0500 5, 30 -- Received: from deliverator8.gatech.edu (localhost [127.0.0.1]) 5, 30 -- by localhost (Postfix) with SMTP id 10BE18F 5, 30 -- for {microscopy-at-microscopy.com} ; Fri, 11 May 2007 23:20:35 -0400 (EDT) 5, 30 -- (envelope-from yb4-at-mail.gatech.edu) 5, 30 -- Received: from webmail4.gatech.edu (webmail4.prism.gatech.edu [130.207.171.134]) 5, 30 -- by deliverator8.gatech.edu (Postfix) with ESMTP id F1D4860 5, 30 -- for {microscopy-at-microscopy.com} ; Fri, 11 May 2007 23:20:34 -0400 (EDT) 5, 30 -- (envelope-from yb4-at-mail.gatech.edu) 5, 30 -- Received: from localhost (localhost [127.0.0.1]) 5, 30 -- by webmail4.gatech.edu (Postfix) with ESMTP id D4EDE2077 5, 30 -- for {microscopy-at-microscopy.com} ; Fri, 11 May 2007 23:20:34 -0400 (EDT) 5, 30 -- (envelope-from yb4-at-mail.gatech.edu) 5, 30 -- Received: from adsl-75-3-15-50.dsl.chcgil.sbcglobal.net (adsl-75-3-15-50.dsl.chcgil.sbcglobal.net [75.3.15.50]) 5, 30 -- by webmail.mail.gatech.edu (IMP) with HTTP 5, 30 -- for {yb4-at-train.mail.gatech.edu} ; Fri, 11 May 2007 23:20:34 -0400 5, 30 -- Message-ID: {1178940034.46453282c7212-at-webmail.mail.gatech.edu} 5, 30 -- Date: Fri, 11 May 2007 23:20:34 -0400 5, 30 -- From: Yolande Berta {yb4-at-mail.gatech.edu} 5, 30 -- To: microscopy-at-microscopy.com 5, 30 -- Subject: focusing screen on JEOL 100CX II 5, 30 -- MIME-Version: 1.0 5, 30 -- Content-Type: text/plain; charset=ISO-8859-1 5, 30 -- Content-Transfer-Encoding: 8bit 5, 30 -- User-Agent: Internet Messaging Program (IMP) 3.2.5 5, 30 -- X-Originating-IP: 75.3.15.50 5, 30 -- X-Authenticated-User: yb4 ==============================End of - Headers==============================
We could use some suggestions/collective experience about solid substrates for doing SEM imaging of dispersed small particles.
What we are imaging: silicate and oxide mineral grains and silicate glass particles down to the 0.1 micron size range (maybe smaller). Objective is to disperse the particle on a substrate and do total particle counting and automated measuring without "losing" the smallest particles from the data set.
What we need: A substrate for dispersing the particles that would fit two requirements: 1) have a pretty low average Z (carbon would be ideal but I'm willing to consider other ideas), so that my particles will stand out strongly in BSE, and then 2) be as smooth as possible on the sub-micron scale. This last requirement is the tricky one because although there are lots of carbon substrates available commercially, so far we haven't found any that don't have sub-micron scratches and other roughness such that particles in the 0.1 micron size range don't get lost in the surface topography.
Well, that's about all I can think of. Thanks to all of you in advance.
Roy Christoffersen FE-STEM Facility Manager ARES Directorate NASA Johnson Space Center, Houston, TX rcsaic-at-sbcglobal.net
==============================Original Headers============================== 10, 24 -- From rcsaic-at-sbcglobal.net Mon May 14 08:29:19 2007 10, 24 -- Received: from smtp114.sbc.mail.mud.yahoo.com (smtp114.sbc.mail.mud.yahoo.com [68.142.198.213]) 10, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l4EDTIqY011500 10, 24 -- for {Microscopy-at-microscopy.com} ; Mon, 14 May 2007 08:29:19 -0500 10, 24 -- Message-Id: {200705141329.l4EDTIqY011500-at-ns.microscopy.com} 10, 24 -- Received: (qmail 51458 invoked from network); 14 May 2007 13:29:18 -0000 10, 24 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 10, 24 -- s=s1024; d=sbcglobal.net; 10, 24 -- h=Received:X-YMail-OSG:From:To:Subject:Date:MIME-Version:Content-Type:Content-Transfer-Encoding:X-Mailer:Thread-Index:X-MimeOLE; 10, 24 -- b=M/6g2fasyr9sJvhlcrAZdv8JJjEBs45sUMviE9v4eg3YKap5vROm8D1FkJxZbbHDQ1M9a/7Ni6cFYougsFxS3ktxVACaPqdi+j0oVeI6jkP51iUt/rOq+wkl1WFjxnQAG4LgW92mCvbCRnkk41AyLtpTD5CI29LSneizgbb9WEU= ; 10, 24 -- Received: from unknown (HELO STUDYDESKTOP) (rcsaic-at-sbcglobal.net-at-65.69.0.179 with login) 10, 24 -- by smtp114.sbc.mail.mud.yahoo.com with SMTP; 14 May 2007 13:29:18 -0000 10, 24 -- X-YMail-OSG: L0bdRMAVM1mmitUT9y78tG1lrhpYKeX1mEb6uuZtFP.ulSaU6FckuWN0oDcMgMHq3EYfaPNe1w-- 10, 24 -- From: "Roy Christoffersen" {rcsaic-at-sbcglobal.net} 10, 24 -- To: {Microscopy-at-microscopy.com} 10, 24 -- Subject: SEM-help finding smooth, low-Z substrates 10, 24 -- Date: Mon, 14 May 2007 08:29:17 -0500 10, 24 -- MIME-Version: 1.0 10, 24 -- Content-Type: text/plain; 10, 24 -- charset="us-ascii" 10, 24 -- Content-Transfer-Encoding: 7bit 10, 24 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 10, 24 -- Thread-Index: AceWK971Nh/2wWHeQviipHHcsxZtPA== 10, 24 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 ==============================End of - Headers==============================
Neal, Scott, I think I may have missed a couple of posts on this thread, but I'd like to clarify my comments... There were two things - first, that three days to get a sample by ion milling is pretty unusual, and doesn't reflect my experience with the PIPS. Second, the angle of incidence is another parameter in addition to beam energy which you can vary and might give a better or more efficient way of making samples.
If you assume that the heating of the sample is due to the kinetic energy of the ion beam, and that the most important component of the beam's velocity is that which is perpendicular to the sample surface, then heating of the sample should roughly increase with the square of the angle of incidence (using E=0.5mv^2 and sin(theta)=theta for small angles). So you can reduce the heating of the sample about 25 times by dropping from 5 degrees incidence angle to 1 degree - which is much more efficient than reducing the beam energy by 25 times. The milling rate doesn't drop off as quickly as the reduction in heating. At least this is my way of explaining why I melt my InP specimens at 6 degrees incidence angle but get better results than my old ion mill (which had LN2 cooling and minimum 10 degrees incidence angle) using 2 degrees in the PIPS.
Having said all this, I don't work with glass samples and if you have a way of making good samples which is sucessful, that's a good thing (finding a protocol which works can be hard enough!).
-----Original Message----- X-from: walck-at-southbaytech.com [mailto:walck-at-southbaytech.com] Sent: 11 May 2007 18:44 To: Richard Beanland
I'd like to contribute a little to the ion milling damage issue being discussed. I think that I have used every brand of ion mill on the market in my career at some point or another. When I was with PPG, glass substrates were the only cross section samples that I dealt with and they were the most tricky. With those samples, I had the most trouble with the PIPS(TM) instrument. If you look at the milling rates with other instruments, the PIPS(TM) Penning ion guns are similar in ion sputtering rates to other ion guns in other companies' ion mills (except the Technoorg-Linda gun with the Einzel focusing lens). It is actually very hard to compare them because everyone does it differently. I believe that the issue wasn't one of the ion beam, but so for the design of the clamp-type DuoPost(TM) that I was using to mill at low angles. The DuoPost(TM) does not have a large mass, and the gripping method does not have a great thermal path to extract heat away from the sample. Glass also doesn't have a great thermal conductivity. If I did not operate the PIPS with lowered ion gun performance parameters, I would soften my glass samples and they would soften and "droop down" during milling. This did not occur with any other ion mill that I used that had a more massive sample holder with better heat path. These other mills included the ones from BalTec (used extensively), EA Fischione (used extensively), and Technoorg-Linda (only a few samples).
With respect to ion damage from ion milling, I think that Arpad Barna's group's work is definitive on the topic. He ion milled Si and GaAs samples at different energies at 5 degrees and then immediately evaporated Al on top to protect the samples from oxidation when they were taken out of the ion mill. He then prepared cross sections of those samples and measured the thickness of the amorphous damage region on the surface due to the ion milling by using HREM. For Si, they saw that at 3 kV (about the limit for conventional ion guns) an amorphous region 120 Angstroms was formed. Using the Technoorg-Linda low energy gun at 250 eV, they achieved an amorphous region of about 10 Angstroms. (A. Barna, B. Pecz; Amorphisation and surface morphology development at low-energy ion milling; Ultramicroscopy 70, 1998.) In other studies and examples, they have also shown that the mottling from ion milled and FIB samples using the low energy guns can be completely eliminated. I have some of the data from that work if you are interested in seeing the results.
Disclaimer: South Bay Technology, Inc. is the exclusive distributer for Technoorg-Linda products in the United States.
-Scott
Scott D. Walck, Ph.D. Technical Director South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
US Toll Free: 1-800-728-2233 Tel: (949) 492-2600 Fax: (949) 492-1499
www.southbaytech.com walck-at-southbaytech.com
-----Original Message----- X-from: nealzimm-at-cpinternet.com [mailto:nealzimm-at-cpinternet.com] Sent: Friday, May 11, 2007 5:53 AM To: Walck-at-SouthBayTech.com
Richard, Witold;
I'll try to reply to both your questions with this one post.
Because I use and maintain PIPS does not make me an "expert" at ion milling. I have noticed a few things that do/do not happen under the different milling conditions we work with.
Low kv milling seems to give lower instances of twinning and stacking faults in the thin films I have worked with. But I do not think that milling damage goes away at lower kVs. I see a background of "mottling" in the films I have imaged. This is what I assume is the damage from milling and occurs at 5kV milling as well as 2.6kV milling.
The milling times are so long for the reason that most of the thin-films-on-glass (silicon) are created to study the behavior as deposited - no anneal. This makes the films fragile to the point that even the technique of encasing the sample in a brass tube does not work - the glue bond bonding the coupons together is greater than the bonding of the film to the substrate.
For mechanically prepared films (which our lead microscopist prefers over FIB cut samples) I had to come up with a way of bonding a glass cover slip to the substrate and trapping the film between. As has been mentioned here, glass behaves poorly in an ion beam. As it becomes very thin, it melts and (usually) pulls the epoxy and film off of the substrate.
The only way to stop the melting is to go to low kVs and a low duty cycle. For most samples this is 3.5kV and 1 minute of milling for every 5 minutes. And as I am only around for 8 hours a day, this makes for a 3 day milling session.
I have been able to prepare sections of Pb (actually PbZrTiOx) that give spectacular lattice (although disappointingly similar to silicon). As far as reducing the time for milling silicon samples, at ten microns thickness and 3kV milling on non-temperature sensitive samples, milling times are usually in the 15 to 20 minute range.
Neal
On Thu, 2007-05-10 at 09:10 -0500, richard.beanland-at-bookham.com wrote: } } } ---------------------------------------------------------------------- } ------ The Microscopy ListServer -- CoSponsor: The Microscopy Society
} of America To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } Neal, } I wouldn't expect a 10um Si sample to take more than 30-60 mins to } thin in a PIPS. } I can only think you are working at a relatively high milling angle } (} 5 degrees) and so have to turn the beam energy down very low so you } don't damage your sample. Why not experiment with one sample using } full power (6kV) and 3 degrees incidence angle, finishing off with 10 } mins at 2kV? It could increase your throughput more than 20 times! } } Richard } } } } ________________________________________ } Richard Beanland } Materials Analysis } Bookham } Caswell } Towcester } Northants } NN12 8EQ } United Kingdom } Tel. +44 1327 356362 } Fax. +44 1327 356775 } http://www.bookham.com } ________________________________________ } } } -----Original Message----- } X-from: nealzimm-at-cpinternet.com [mailto:nealzimm-at-cpinternet.com] } Sent: 10 May 2007 14:10 } To: Richard Beanland } Subject: [Microscopy] Re: Looking for recommendations about dual beam } ion } } } } } ---------------------------------------------------------------------- } -- } ---- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -- } ---- } } John } } I work with four Gatan PIPS daily and have found them to be quite } acceptable in ease of both maintenance and use. } } We currently do maintenance only when the ion guns stop functioning, } at that point the machine is taken out of service and a full cleaning } performed. This happens about every five to six months and leaves the
} PIPS out of service for two days (One day to clean, pump down over } night, and alignment the next day). The diaphragm pump needs new } diaphragms once a year (about an hours work) but not too much else. } } Work load is typically one to two ceramic samples per day, with an } occasional silicon or glass sample thrown in. The ceramic samples are
} lapped to about six microns and mill for an hour to two hours. } Silicon and glass are left about ten microns thick and take much } longer (up to three days), not so much due to the thickness as to the } low kVs used and letting the sample "cool". } } Gatan offers a digital camera/zoom lens set and an external diaphragm } pump as extras. In my opinion, they are a "must have". } } If you have any further questions please feel free to contact me. } } Neal R. Zimmermann } } } } On Wed, 2007-05-09 at 15:43 -0500, jpchandl-at-mines.edu wrote: } } } } } } } ---------------------------------------------------------------------- } -- } ---- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } ---------------------------------------------------------------------- } -- } ---- } } } } I need to tap in to the wisdom of the list about dual beam argon ion } mills. } } } } We have a Bal-Tec RES100 ion mill that we are having difficulty } maintaining. } } It had been in disuse for a couple of years, and has been repaired } } and upgraded recently, at the factory, to be as close to a RES101 as } possible. } } We are having issues such as the guns becoming contaminated and } needing } } service much more frequently than we think they should. Routine } maintenance } } seems to be difficult, too. We think we are looking at a high } maintenance } } instrument here. I would like to know what kind of experience other } labs } } have had with this instrument. } } } } I would also like to hear from people who have experience } using/maintaining } } the RES100/101 or the Gatan PIPS, or both. We need some comparative
} } information, so we can make a decision about how to proceed with our } ion } } milling needs. } } } } Any input would be appreciated. } } } } --John } } } } John Chandler } } Manager, EM Lab } } Colorado School of Mines } } Golden, CO 80401 } } jpchandl-at-mines.edu } } 303-384-2203 } } } } } } } } } } ==============================Original } Headers============================== } } 9, 19 -- From jpchandl-at-mines.edu Wed May 9 15:42:15 2007 9, 19 -- } } Received: from inspire.Mines.EDU (inspire.Mines.EDU } [138.67.130.5]) } } 9, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id l49KgEDO003975 } } 9, 19 -- for {microscopy-at-microscopy.com} ; Wed, 9 May 2007 } 15:42:14 -0500 } } 9, 19 -- Received: from emlab1 (emlab1.Mines.EDU [138.67.28.173]) } } 9, 19 -- by inspire.Mines.EDU (8.13.1/8.13.1) with ESMTP id } l49KgD7T016786 } } 9, 19 -- for {microscopy-at-microscopy.com} ; Wed, 9 May 2007 } 14:42:13 -0600 } } 9, 19 -- From: "John Chandler" {jpchandl-at-mines.edu} 9, 19 -- To: } } {microscopy-at-microscopy.com} 9, 19 -- Subject: Looking for } } recommendations about dual beam ion } mills } } 9, 19 -- Date: Wed, 9 May 2007 14:42:13 -0600 9, 19 -- Message-ID: } } {001e01c7927a$861c8570$ad1c438a-at-mines.edu} } } 9, 19 -- MIME-Version: 1.0 } } 9, 19 -- Content-Type: text/plain; } } 9, 19 -- charset="us-ascii" } } 9, 19 -- Content-Transfer-Encoding: 7bit 9, 19 -- X-Mailer: } } Microsoft Office Outlook 11 9, 19 -- X-MimeOLE: Produced By } } Microsoft MimeOLE V6.00.2900.3028 9, 19 -- Thread-Index: } } AceSeoX7iyPEBSG3T3O4yAVZe0waUw== ==============================End } } of - } Headers============================== } } } ==============================Original } Headers============================== } 11, 24 -- From nealzimm-at-cpinternet.com Thu May 10 08:08:28 2007 11, 24
} -- Received: from cpinternet.com (mail.cpinternet.com } [209.240.224.81]) } 11, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id l4AD8S9W001596 } 11, 24 -- for {microscopy-at-microscopy.com} ; Thu, 10 May 2007 } 08:08:28 -0500 } 11, 24 -- Received: from [192.168.1.102] (unverified [216.251.191.121]) } 11, 24 -- by cpinternet.com (SurgeMail 3.8j) with ESMTP id } 171217060-1881964 } 11, 24 -- for multiple; Thu, 10 May 2007 08:08:27 -0500 CDT } 11, 24 -- Subject: Re: [Microscopy] Looking for recommendations about } dual beam ion } 11, 24 -- mills } 11, 24 -- From: Neal Zimmermann {nealzimm-at-cpinternet.com} 11, 24 -- } To: jpchandl-at-mines.edu, microscopy-at-microscopy.com 11, 24 -- } In-Reply-To: {200705092043.l49Kh2W8005876-at-ns.microscopy.com} } 11, 24 -- References: {200705092043.l49Kh2W8005876-at-ns.microscopy.com} } 11, 24 -- Content-Type: text/plain } 11, 24 -- Date: Thu, 10 May 2007 08:08:14 -0500 11, 24 -- Message-Id: } {1178802494.3663.19.camel-at-localhost.localdomain} } 11, 24 -- Mime-Version: 1.0 } 11, 24 -- X-Mailer: Evolution 2.8.3 (2.8.3-2.fc6) 11, 24 -- } Content-Transfer-Encoding: 7bit 11, 24 -- X-Authenticated-User: } nealzimm-at-cpinternet.com 11, 24 -- X-NotAscii: charset=us-ascii 11, 24 } -- X-Encryption: SSL encrypted 11, 24 -- X-IP-stats: No info recorded } yet ip=216.251.191.121 11, 24 -- X-Originating-IP: 216.251.191.121 } ==============================End of - } Headers============================== } } ====================================================================== } = This e-mail is intended for the person it is addressed to only. The } information contained in it may be confidential and/or protected by } law. If you are not the intended recipient of this message, you must } not make any use of this information, or copy or show it to any } person. Please contact us immediately to tell us that you have } received this e-mail, and return the original to us. Any use, } forwarding, printing or copying of this message is strictly prohibited. } No part of this message can be considered a request for goods or } services. } ====================================================================== } = } } } ==============================Original } Headers============================== } 23, 34 -- From richard.beanland-at-bookham.com Thu May 10 09:09:28 2007 } 23, 34 -- Received: from mail72.messagelabs.com (mail72.messagelabs.com [193.109.255.147]) } 23, 34 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l4AE9Riq025754 } 23, 34 -- for {microscopy-at-microscopy.com} ; Thu, 10 May 2007 09:09:27 -0500 } 23, 34 -- X-VirusChecked: Checked } 23, 34 -- X-Env-Sender: richard.beanland-at-bookham.com 23, 34 -- } X-Msg-Ref: server-10.tower-72.messagelabs.com!1178806165!33452822!1 } 23, 34 -- X-StarScan-Version: 5.5.10.7.1; banners=bookham.com,-,- 23, } 34 -- X-Originating-IP: [213.249.209.179] 23, 34 -- Received: (qmail } 31853 invoked from network); 10 May 2007 14:09:26 -0000 23, 34 -- } Received: from unknown (HELO cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI) (213.249.209.179) } 23, 34 -- by server-10.tower-72.messagelabs.com with SMTP; 10 May 2007 14:09:26 -0000 } 23, 34 -- Received: from cas-smx-02.BOOKHAM.ENTERPRISE.PRI ([10.4.0.223]) by cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI with Microsoft SMTPSVC(6.0.3790.1830); } 23, 34 -- Thu, 10 May 2007 15:10:39 +0100 } 23, 34 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 23, 34 -- } Content-class: urn:content-classes:message 23, 34 -- MIME-Version: 1.0
==============================Original Headers============================== 11, 22 -- From nealzimm-at-cpinternet.com Fri May 11 07:47:04 2007 11, 22 -- Received: from cpinternet.com (mail.cpinternet.com [209.240.224.81]) 11, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4BCl3QP022768 11, 22 -- for {microscopy-at-microscopy.com} ; Fri, 11 May 2007 07:47:03 -0500 11, 22 -- Received: from [192.168.1.102] (unverified [216.251.190.54]) 11, 22 -- by cpinternet.com (SurgeMail 3.8j) with ESMTP id 172671156-1881964 11, 22 -- for {microscopy-at-microscopy.com} ; Fri, 11 May 2007 07:47:03 -0500 CDT 11, 22 -- Subject: Re: [Microscopy] Looking for recommendations about dual beam ion 11, 22 -- From: Neal Zimmermann {nealzimm-at-cpinternet.com} 11, 22 -- To: microscopy-at-microscopy.com 11, 22 -- In-Reply-To: {200705101410.l4AEAngB027743-at-ns.microscopy.com} 11, 22 -- References: {200705101410.l4AEAngB027743-at-ns.microscopy.com} 11, 22 -- Content-Type: text/plain 11, 22 -- Date: Fri, 11 May 2007 07:46:51 -0500 11, 22 -- Message-Id: {1178887611.5048.22.camel-at-localhost.localdomain} 11, 22 -- Mime-Version: 1.0 11, 22 -- X-Mailer: Evolution 2.8.3 (2.8.3-2.fc6) 11, 22 -- Content-Transfer-Encoding: 7bit 11, 22 -- X-Authenticated-User: nealzimm-at-cpinternet.com 11, 22 -- X-Encryption: SSL encrypted 11, 22 -- X-IP-stats: Incoming Last 2, First 1, in=4, out=0, spam=0 ip=216.251.190.54 11, 22 -- X-Originating-IP: 216.251.190.54 ==============================End of - Headers==============================
==============================Original Headers============================== 22, 22 -- From walck-at-southbaytech.com Fri May 11 12:42:35 2007 22, 22 -- Received: from flpi102.sbcis.sbc.com (flpi102.sbcis.sbc.com [207.115.20.71]) 22, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4BHgZrx005622 22, 22 -- for {Microscopy-at-microscopy.com} ; Fri, 11 May 2007 12:42:35 -0500 22, 22 -- X-ORBL: [64.169.217.123] 22, 22 -- Received: from dynamicbl8uno3 (adsl-64-169-217-123.dsl.lsan03.pacbell.net [64.169.217.123]) 22, 22 -- by flpi102.sbcis.sbc.com (8.13.8 out.dk.spool/8.13.8) with ESMTP id l4BHgLVU020749; 22, 22 -- Fri, 11 May 2007 10:42:22 -0700 22, 22 -- From: "Scott Walck" {walck-at-southbaytech.com} 22, 22 -- To: {Microscopy-at-microscopy.com} 22, 22 -- Cc: {nealzimm-at-cpinternet.com} 22, 22 -- Subject: RE: [Microscopy] Re: Looking for recommendations about dual beam ion 22, 22 -- Date: Fri, 11 May 2007 10:42:28 -0700 22, 22 -- Message-ID: {002d01c793f3$bf1292f0$7801a8c0-at-dynamicbl8uno3} 22, 22 -- MIME-Version: 1.0 22, 22 -- Content-Type: text/plain; 22, 22 -- charset="us-ascii" 22, 22 -- Content-Transfer-Encoding: 7bit 22, 22 -- X-Mailer: Microsoft Office Outlook 11 22, 22 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 22, 22 -- In-Reply-To: {200705111252.l4BCqYea028524-at-ns.microscopy.com} 22, 22 -- Thread-Index: AceTyz6GLfgJgdXUQD604ooQaJ5lGQAIEt2g ==============================End of - Headers==============================
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==============================Original Headers============================== 42, 34 -- From richard.beanland-at-bookham.com Mon May 14 09:07:35 2007 42, 34 -- Received: from mail80.messagelabs.com (mail80.messagelabs.com [195.245.230.163]) 42, 34 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l4EE7YrH023998 42, 34 -- for {microscopy-at-microscopy.com} ; Mon, 14 May 2007 09:07:34 -0500 42, 34 -- X-VirusChecked: Checked 42, 34 -- X-Env-Sender: richard.beanland-at-bookham.com 42, 34 -- X-Msg-Ref: server-11.tower-80.messagelabs.com!1179151652!42754681!1 42, 34 -- X-StarScan-Version: 5.5.10.7.1; banners=bookham.com,-,- 42, 34 -- X-Originating-IP: [213.249.209.179] 42, 34 -- Received: (qmail 7030 invoked from network); 14 May 2007 14:07:33 -0000 42, 34 -- Received: from unknown (HELO cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI) (213.249.209.179) 42, 34 -- by server-11.tower-80.messagelabs.com with SMTP; 14 May 2007 14:07:33 -0000 42, 34 -- Received: from cas-smx-02.BOOKHAM.ENTERPRISE.PRI ([10.4.0.223]) by cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI with Microsoft SMTPSVC(6.0.3790.1830); 42, 34 -- Mon, 14 May 2007 15:08:48 +0100 42, 34 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 42, 34 -- Content-class: urn:content-classes:message 42, 34 -- MIME-Version: 1.0 42, 34 -- Content-Type: text/plain; 42, 34 -- charset="us-ascii" 42, 34 -- Subject: RE: [Microscopy] Looking for recommendations about dual beam ion 42, 34 -- Date: Mon, 14 May 2007 15:08:47 +0100 42, 34 -- Message-ID: {9645D3E33E4C6548B12A7B25F611533E46CA81-at-cas-smx-02.BOOKHAM.ENTERPRISE.PRI} 42, 34 -- In-Reply-To: {200705111743.l4BHhVYJ007595-at-ns.microscopy.com} 42, 34 -- X-MS-Has-Attach: 42, 34 -- X-MS-TNEF-Correlator: 42, 34 -- Thread-Topic: [Microscopy] Looking for recommendations about dual beam ion 42, 34 -- Thread-Index: AceT9B0D8Z/fqVm1Qdq71wHdAeJAPwCOlqTA 42, 34 -- References: {200705111743.l4BHhVYJ007595-at-ns.microscopy.com} 42, 34 -- From: "Richard Beanland" {richard.beanland-at-bookham.com} 42, 34 -- To: {walck-at-southbaytech.com} , {nealzimm-at-cpinternet.com} 42, 34 -- Cc: {microscopy-at-microscopy.com} 42, 34 -- X-OriginalArrivalTime: 14 May 2007 14:08:48.0648 (UTC) FILETIME=[6477B880:01C79631] 42, 34 -- Content-Transfer-Encoding: 8bit 42, 34 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4EE7YrH023998 ==============================End of - Headers==============================
How about using a TEM carbon support film? Disperse your particles on the support film then mount the grid on a "Faraday cup"
I made a TEM grid support by taking about a 1cm long piece of 1/4" graphite rod, drilling out the center 6-7mm deep, and using a 1/8" (3.1mm) end mill to make a small recess for the grid in the end of the rod. I used carbon dag to glue the rod upright on an SEM stub and presto... instant TEM grid holder with near zero background!
Cheers, Henk
At 09:32 AM 05/14/07, you wrote:
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://www.ceof.ohio-state.edu Time is that quality of nature which keeps events from happening all at once. Lately it doesn't seem to be working.
==============================Original Headers============================== 10, 26 -- From colijn.1-at-osu.edu Mon May 14 09:26:18 2007 10, 26 -- Received: from ER6S1.ENG.OHIO-STATE.EDU (er6s1.ecr6.ohio-state.edu [164.107.76.2]) 10, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4EEQHns003274 10, 26 -- for {microscopy-at-microscopy.com} ; Mon, 14 May 2007 09:26:18 -0500 10, 26 -- Received: from CONVERSION-DAEMON.er6s1.eng.ohio-state.edu by 10, 26 -- er6s1.eng.ohio-state.edu (PMDF V6.2-1x11 #31056) 10, 26 -- id {01MGKMOJSKHCA9S5A4-at-er6s1.eng.ohio-state.edu} for 10, 26 -- microscopy-at-microscopy.com; Mon, 14 May 2007 10:26:15 -0400 (EDT) 10, 26 -- Received: from HOC1.ecr6.ohio-state.edu 10, 26 -- (hoc1.ceof.ohio-state.edu [164.107.76.179]) by er6s1.eng.ohio-state.edu 10, 26 -- (PMDF V6.2-1x11 #31056) 10, 26 -- with ESMTPA id {01MGKMOJBPXQA9VIVU-at-er6s1.eng.ohio-state.edu} ; Mon, 10, 26 -- 14 May 2007 10:26:14 -0400 (EDT) 10, 26 -- Date: Mon, 14 May 2007 10:26:49 -0400 10, 26 -- From: "Hendrik O. Colijn" {colijn.1-at-osu.edu} 10, 26 -- Subject: Re: [Microscopy] SEM-help finding smooth, low-Z substrates 10, 26 -- In-reply-to: {200705141332.l4EDWGNm014144-at-ns.microscopy.com} 10, 26 -- Sender: colijn-at-er6s1.ecr6.ohio-state.edu 10, 26 -- To: rcsaic-at-sbcglobal.net 10, 26 -- Cc: microscopy-at-microscopy.com 10, 26 -- Message-id: {7.0.1.0.2.20070514102004.03842288-at-osu.edu} 10, 26 -- MIME-version: 1.0 10, 26 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 10, 26 -- Content-type: text/plain; charset=us-ascii; format=flowed 10, 26 -- X-Env-From: auth/colijn.1-at-osu.edu 10, 26 -- References: {200705141332.l4EDWGNm014144-at-ns.microscopy.com} ==============================End of - Headers==============================
} Caleb Adams and I have been carrying on a conversation about training and jobs in microscopy. Can any of you provide him with further information? Since Caleb is not on the listserver, please respond to him directly (Caleb Adams {cannon_adamscl-at-yahoo.com} )
Thanks, Barbara Foster, President
Microscopy/Microscopy Education 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 W: www.MicroscopyEducation.com
MME is now scheduling customized, on-site courses through July. Call us today for details.
P. S. Need a good general reference or light microscopy text for next semester? Call us today to learn more about "Optimizing LIght Microscopy". Copies still available through MME... even for class-room lots ... and we give quantity discounts. Call Ken Piel at (972)954-8011.
} Barbara, } } Thank you for the thorough info. I have been working } with my Leica CME microscope for about 4 months and } wanted to know all the equipment that would be good to } have with my microscope. I currently have the } following accessories: slides and cover slips, } dissecting tool set, darkfield, cleaner, slide and } microscope case, and petri dish. I will be done with } my high school within 1 to 2 years and from there } would like to work in a non medical field of } microscopy but in the meantime is there some online } course in microscopy that I could take? } } Sincerely, } Caleb } --- Barbara Foster {bfoster-at-mme1.com} wrote: } } } Hi, Caleb } } } } That's a pretty hard call to make because there are } } so many factors. } } 1. Where is microscopy done? } } Microscopy is used universally, but your best bet } } would be in } } industry or as a lab tech in a university's central } } service lab. } } 2. What certifications are required? } } If you do microscopy in a clinical lab, you need to } } have the training } } and certification to be a lab tech. I think that } } that is usually } } either a 2 year or 4 year program. In industry, a } } lot of training is } } done "on the job", so if you have a basic } } understanding of science } } (the simple physics of optics is important... you } } can buy a second } } hand high school physics book or a review book. } } Also, our book, } } "Optimizing Light Microscopy" has what you need) and } } are willing to } } start low and work your way up, that's a possibility } } 3. What type of microscopy? } } It would probably be easiest to start with light } } (optical) } } microscopy, then expand in to atomic force or } } electron microscopy. } } 4. Where to find jobs? } } I'd start traveling around the industrial parks in } } the neighborhood } } and trying to identify businesses that might use } } microscopy for } } quality control. Also, Chem & Engineering News } } (www.acs.org) has a } } job bank... and you might try the traditional road } } on-line } } (Monster.com, etc.). Also, you'd be surprised at } } what your friends } } and family "don't know that they know." Start } } chatting them up, to } } see if they know someone who knows someone who knows } } someone. } } } } I guess that's it. I'd suggest that you plan for } } some advanced } } education along the line, but if you can get into a } } company at an } } entry level, they might help you pay for an } } associates degree or even } } further. Local societies (I don't know where you } } are, but the New } } York Microscopical Society) offer weekend courses } } and some colleges } } such as LeHigh offer 1 week/intensive programs that } } will really give } } you a boost. Also, the Royal Microscopical Society } } in Oxford, } } England offers a whole series of 1 week courses } } (that's how I got } } started). Sounds strange, but sometimes it is less } } expensive to } } travel to the UK than to spend a week at a course } } here! Also, I know } } that San Joaquin Delta College in California has a } } full 2 year } } program that has a great reputation. Dr. Judy Murphy } } runs it.... you } } might want to investigate it further. } } } } I hope that all of this is helpful. } } } } Please let me know if you decide to make this career } } shift. } } Best regards, } } Barbara } } } } At 11:25 AM 5/11/2007, Caleb Adams wrote: } } } Barbara, } } } } } } How hard is it to get a job doing microscopy of } } some } } } sort when all you have is a high school education } } and } } } where would I look to find those jobs? } } } } } } Caleb } } } } } } } } } } } } } ____________________________________________________________________________________ } } } Bored stiff? Loosen up... } } } Download and play hundreds of games for free on } } Yahoo! Games. } } } http://games.yahoo.com/games/front } } } } } } } } } ____________________________________________________________________________________Pinpoint customers who are looking for what you sell. } http://searchmarketing.yahoo.com/
==============================Original Headers============================== 11, 16 -- From bfoster-at-mme1.com Mon May 14 09:49:32 2007 11, 16 -- Received: from mail.plhosting.com (qmail1i.plhosting.com [65.39.254.96]) 11, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4EEnVaH015113 11, 16 -- for {microscopy-at-microscopy.com} ; Mon, 14 May 2007 09:49:31 -0500 11, 16 -- Message-Id: {200705141449.l4EEnVaH015113-at-ns.microscopy.com} 11, 16 -- Received: (qmail 5740 invoked by uid 0); 14 May 2007 14:49:31 -0000 11, 16 -- Received: from unknown (HELO barbsd505.mme1.com) (bfoster-at-mme1.com-at-65.17.57.99) 11, 16 -- by qmail1i.plhosting.com with ESMTPA; 14 May 2007 14:49:30 -0000 11, 16 -- X-Mailer: QUALCOMM Windows Eudora Version 7.1.0.9 11, 16 -- Date: Mon, 14 May 2007 09:48:59 -0500 11, 16 -- To: microscopy-at-microscopy.com, Caleb Adams {cannon_adamscl-at-yahoo.com} 11, 16 -- From: Barbara Foster {bfoster-at-mme1.com} 11, 16 -- Subject: Posting for non-MSA member Re: Microscope Job - Please respond 11, 16 -- diretly 11, 16 -- Mime-Version: 1.0 11, 16 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
Can you use a high-Z background rather than low? That should give you dark on light in place of light on dark...
If so, you might want to investigate metal-glass braze foil. Typically it contains Si, Cr, Fe, Ni, or some combination (don't have the specs handy). Since it is a glassy metal, there is no grain structure to interfere with BSE imaging and the surface is relatively smooth. There are some undulations/waves in the surface formed when frozen from the molten state, but maybe not too much to interfere.
Woody White BWXT Services
-----Original Message----- X-from: rcsaic-at-sbcglobal.net [mailto:rcsaic-at-sbcglobal.net] Sent: Monday, May 14, 2007 9:31 AM To: White, Woody N.
Dear Listers,
We could use some suggestions/collective experience about solid substrates for doing SEM imaging of dispersed small particles.
What we are imaging: silicate and oxide mineral grains and silicate glass particles down to the 0.1 micron size range (maybe smaller). Objective is to disperse the particle on a substrate and do total particle counting and automated measuring without "losing" the smallest particles from the data set.
What we need: A substrate for dispersing the particles that would fit two requirements: 1) have a pretty low average Z (carbon would be ideal but I'm willing to consider other ideas), so that my particles will stand out strongly in BSE, and then 2) be as smooth as possible on the sub-micron scale. This last requirement is the tricky one because although there are lots of carbon substrates available commercially, so far we haven't found any that don't have sub-micron scratches and other roughness such that particles in the 0.1 micron size range don't get lost in the surface topography.
Well, that's about all I can think of. Thanks to all of you in advance.
Roy Christoffersen FE-STEM Facility Manager ARES Directorate NASA Johnson Space Center, Houston, TX rcsaic-at-sbcglobal.net
==============================Original Headers============================== 10, 24 -- From rcsaic-at-sbcglobal.net Mon May 14 08:29:19 2007 10, 24 -- Received: from smtp114.sbc.mail.mud.yahoo.com (smtp114.sbc.mail.mud.yahoo.com [68.142.198.213]) 10, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l4EDTIqY011500 10, 24 -- for {Microscopy-at-microscopy.com} ; Mon, 14 May 2007 08:29:19 -0500 10, 24 -- Message-Id: {200705141329.l4EDTIqY011500-at-ns.microscopy.com} 10, 24 -- Received: (qmail 51458 invoked from network); 14 May 2007 13:29:18 -0000 10, 24 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 10, 24 -- s=s1024; d=sbcglobal.net; 10, 24 -- h=Received:X-YMail-OSG:From:To:Subject:Date:MIME-Version:Content-Type:Co ntent-Transfer-Encoding:X-Mailer:Thread-Index:X-MimeOLE; 10, 24 -- b=M/6g2fasyr9sJvhlcrAZdv8JJjEBs45sUMviE9v4eg3YKap5vROm8D1FkJxZbbHDQ1M9a/ 7Ni6cFYougsFxS3ktxVACaPqdi+j0oVeI6jkP51iUt/rOq+wkl1WFjxnQAG4LgW92mCvbCRn kk41AyLtpTD5CI29LSneizgbb9WEU= ; 10, 24 -- Received: from unknown (HELO STUDYDESKTOP) (rcsaic-at-sbcglobal.net-at-65.69.0.179 with login) 10, 24 -- by smtp114.sbc.mail.mud.yahoo.com with SMTP; 14 May 2007 13:29:18 -0000 10, 24 -- X-YMail-OSG: L0bdRMAVM1mmitUT9y78tG1lrhpYKeX1mEb6uuZtFP.ulSaU6FckuWN0oDcMgMHq3EYfaPNe 1w-- 10, 24 -- From: "Roy Christoffersen" {rcsaic-at-sbcglobal.net} 10, 24 -- To: {Microscopy-at-microscopy.com} 10, 24 -- Subject: SEM-help finding smooth, low-Z substrates 10, 24 -- Date: Mon, 14 May 2007 08:29:17 -0500 10, 24 -- MIME-Version: 1.0 10, 24 -- Content-Type: text/plain; 10, 24 -- charset="us-ascii" 10, 24 -- Content-Transfer-Encoding: 7bit 10, 24 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 10, 24 -- Thread-Index: AceWK971Nh/2wWHeQviipHHcsxZtPA== 10, 24 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 ==============================End of - Headers==============================
==============================Original Headers============================== 21, 28 -- From nwwhite-at-bwxt.com Mon May 14 09:55:16 2007 21, 28 -- Received: from bwxt.com (lynsmtp1.bwxt.com [131.184.146.8]) 21, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4EEtFIp022787 21, 28 -- for {microscopy-at-msa.microscopy.com} ; Mon, 14 May 2007 09:55:15 -0500 21, 28 -- Received: from ([131.184.13.224]) 21, 28 -- by lynsmtp1.bwxt.com with ESMTP id 5202488.4866904; 21, 28 -- Mon, 14 May 2007 10:54:55 -0400 21, 28 -- Received: from BWXSPO01.BWXS.BWXTECH.NET ([131.184.13.31]) by bwxslynbh01.BWXS.BWXTECH.NET with Microsoft SMTPSVC(6.0.3790.1830); 21, 28 -- Mon, 14 May 2007 10:54:55 -0400 21, 28 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 21, 28 -- Content-class: urn:content-classes:message 21, 28 -- MIME-Version: 1.0 21, 28 -- Content-Type: text/plain; 21, 28 -- charset="us-ascii" 21, 28 -- Subject: RE: [Microscopy] SEM-help finding smooth, low-Z substrates 21, 28 -- Date: Mon, 14 May 2007 10:54:55 -0400 21, 28 -- Message-ID: {360DDAABED436B49BA397357CCEE8BB7F87905-at-BWXSPO01.BWXS.BWXTECH.NET} 21, 28 -- In-Reply-To: {200705141330.l4EDUVV8012296-at-ns.microscopy.com} 21, 28 -- X-MS-Has-Attach: 21, 28 -- X-MS-TNEF-Correlator: 21, 28 -- Thread-Topic: [Microscopy] SEM-help finding smooth, low-Z substrates 21, 28 -- Thread-Index: AceWLBKBxGJ3pGwDSWiXH6Hk+fOmKgACsMkg 21, 28 -- From: "White, Woody N." {NWWhite-at-bwxt.com} 21, 28 -- To: {rcsaic-at-sbcglobal.net} , 21, 28 -- "MicroscopyListServer" {Microscopy-at-msa.microscopy.com} 21, 28 -- X-OriginalArrivalTime: 14 May 2007 14:54:55.0549 (UTC) FILETIME=[D5AB4ED0:01C79637] 21, 28 -- Content-Transfer-Encoding: 8bit 21, 28 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4EEtFIp022787 ==============================End of - Headers==============================
Yes, John, it's 8 mm, not microns. The issue is holding the device while making the second cleave or cut. The sample would have to be mounted on something to use a diamond saw, which is available. However, I do need to keep the face of the surface I'm looking at undisturbed with whatever method I use.
Regards,
Peter
Mardinly, John wrote: } ~ 8 mm apart? Did I read this correctly? Not microns? We use a Disco saw } routinely. If it's ~ 8 mm apart, you can use any diamond saw. If it's } microns you need a FIB. } } John Mardinly } Intel Corporation } This email does not represent an opinion of Intel Corporation. } } -----Original Message----- } From: peter.tomic-at-renwireless.com [mailto:peter.tomic-at-renwireless.com] } Sent: Tuesday, May 08, 2007 10:33 AM } To: Mardinly, John } Subject: [Microscopy] Silicon Cross-section sample preparation } } } } } ------------------------------------------------------------------------ } ---- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ } ---- } } Microscopy Folks, } } I'd like to get some input on sample preparation with respect to } silicon. } } PROBLEM: } } I have an FEI XL-50 FESEM that was originally designed to accept flat } samples, essentially silicon wafers. It does not have what one would } consider a typical exchange port. The exchange port is robotically } controlled, and the maximum sample height, including stub, must be 5 mm } or less. I must view these samples on edge, i.e. 90 degrees. This } presents a problem in that I must cleave two parallel sections very } close to each other. Perhaps diamond cutters can do this, but my hands } are too shaky. } } QUESTION: } } Is there a device, or method, that would allow me to make these cleaves } ~ 8 mm apart with any reasonable control? I found stubs that are low } profile, 38 and 90 degrees, from the nice people at Ted Pella, but I } still have this issue of doing two parallel cleaves very close together. } } Just for info. purposes I am in the silicon MEMS development arena. } } If you feel your reply is of general interest to this community, please } reply to all, or you may contact me directly. } } Regards to all in this small world, } } Peter Tomic } Renaissance Wireless Corp. } } }
==============================Original Headers============================== 6, 26 -- From peter.tomic-at-renwireless.com Mon May 14 10:33:20 2007 6, 26 -- Received: from mail.rw.local (4.piok2.xdsl.nauticom.net [209.195.176.101]) 6, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4EFXJSP006437 6, 26 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 14 May 2007 10:33:20 -0500 6, 26 -- Received: from localhost (localhost [127.0.0.1]) 6, 26 -- by mail.rw.local (Postfix) with ESMTP id D2F46246B5; 6, 26 -- Mon, 14 May 2007 11:33:17 -0400 (EDT) 6, 26 -- Received: from mail.rw.local ([127.0.0.1]) 6, 26 -- by localhost (mail.rw.local [127.0.0.1]) (amavisd-new, port 10024) with ESMTP 6, 26 -- id 26580-04; Mon, 14 May 2007 11:33:17 -0400 (EDT) 6, 26 -- Received: from [127.0.0.1] (unknown [10.11.10.110]) 6, 26 -- by mail.rw.local (Postfix) with ESMTP id A2C0918AC7; 6, 26 -- Mon, 14 May 2007 11:33:16 -0400 (EDT) 6, 26 -- Message-ID: {464882B3.7010905-at-renwireless.com} 6, 26 -- Date: Mon, 14 May 2007 11:39:31 -0400 6, 26 -- From: Peter Tomic {peter.tomic-at-renwireless.com} 6, 26 -- User-Agent: Thunderbird 2.0.0.0 (Windows/20070326) 6, 26 -- MIME-Version: 1.0 6, 26 -- To: "Mardinly, John" {john.mardinly-at-intel.com} 6, 26 -- CC: Microscopy-at-MSA.Microscopy.Com 6, 26 -- Subject: Re: [Microscopy] Silicon Cross-section sample preparation 6, 26 -- References: {F3CB8931ABF8294DB889E977150CAD68705D13-at-scsmsx415.amr.corp.intel.com} 6, 26 -- In-Reply-To: {F3CB8931ABF8294DB889E977150CAD68705D13-at-scsmsx415.amr.corp.intel.com} 6, 26 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 6, 26 -- Content-Transfer-Encoding: 7bit 6, 26 -- X-Virus-Scanned: amavisd-new at renwireless.com ==============================End of - Headers==============================
Dear Roy, There is a material called "glassy carbon", which is smooth, solid and can be polished to a mirror surface. It is very hard, but is pure carbon and so it makes a good substrate for BSE studies. Some of the EM catalogue companies used to supply it, they probably still do. I get my glassy carbon planchets (0.5 inch discs) from Canemco (www.canemco.com) and stick them onto my SEM stubs. Good luck,
-----Original Message----- X-from: rcsaic-at-sbcglobal.net [mailto:rcsaic-at-sbcglobal.net] Sent: May 14, 2007 6:38 AM To: mager-at-interchange.ubc.ca
Dear Listers,
We could use some suggestions/collective experience about solid substrates for doing SEM imaging of dispersed small particles.
What we are imaging: silicate and oxide mineral grains and silicate glass particles down to the 0.1 micron size range (maybe smaller). Objective is to disperse the particle on a substrate and do total particle counting and automated measuring without "losing" the smallest particles from the data set.
What we need: A substrate for dispersing the particles that would fit two requirements: 1) have a pretty low average Z (carbon would be ideal but I'm willing to consider other ideas), so that my particles will stand out strongly in BSE, and then 2) be as smooth as possible on the sub-micron scale. This last requirement is the tricky one because although there are lots of carbon substrates available commercially, so far we haven't found any that don't have sub-micron scratches and other roughness such that particles in the 0.1 micron size range don't get lost in the surface topography.
Well, that's about all I can think of. Thanks to all of you in advance.
Roy Christoffersen FE-STEM Facility Manager ARES Directorate NASA Johnson Space Center, Houston, TX rcsaic-at-sbcglobal.net
==============================Original Headers============================== 10, 24 -- From rcsaic-at-sbcglobal.net Mon May 14 08:29:19 2007 10, 24 -- Received: from smtp114.sbc.mail.mud.yahoo.com (smtp114.sbc.mail.mud.yahoo.com [68.142.198.213]) 10, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l4EDTIqY011500 10, 24 -- for {Microscopy-at-microscopy.com} ; Mon, 14 May 2007 08:29:19 -0500 10, 24 -- Message-Id: {200705141329.l4EDTIqY011500-at-ns.microscopy.com} 10, 24 -- Received: (qmail 51458 invoked from network); 14 May 2007 13:29:18 -0000 10, 24 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 10, 24 -- s=s1024; d=sbcglobal.net; 10, 24 -- h=Received:X-YMail-OSG:From:To:Subject:Date:MIME-Version:Content-Type:Conten t-Transfer-Encoding:X-Mailer:Thread-Index:X-MimeOLE; 10, 24 -- b=M/6g2fasyr9sJvhlcrAZdv8JJjEBs45sUMviE9v4eg3YKap5vROm8D1FkJxZbbHDQ1M9a/7Ni6 cFYougsFxS3ktxVACaPqdi+j0oVeI6jkP51iUt/rOq+wkl1WFjxnQAG4LgW92mCvbCRnkk41AyLt pTD5CI29LSneizgbb9WEU= ; 10, 24 -- Received: from unknown (HELO STUDYDESKTOP) (rcsaic-at-sbcglobal.net-at-65.69.0.179 with login) 10, 24 -- by smtp114.sbc.mail.mud.yahoo.com with SMTP; 14 May 2007 13:29:18 -0000 10, 24 -- X-YMail-OSG: L0bdRMAVM1mmitUT9y78tG1lrhpYKeX1mEb6uuZtFP.ulSaU6FckuWN0oDcMgMHq3EYfaPNe1w-- 10, 24 -- From: "Roy Christoffersen" {rcsaic-at-sbcglobal.net} 10, 24 -- To: {Microscopy-at-microscopy.com} 10, 24 -- Subject: SEM-help finding smooth, low-Z substrates 10, 24 -- Date: Mon, 14 May 2007 08:29:17 -0500 10, 24 -- MIME-Version: 1.0 10, 24 -- Content-Type: text/plain; 10, 24 -- charset="us-ascii" 10, 24 -- Content-Transfer-Encoding: 7bit 10, 24 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 10, 24 -- Thread-Index: AceWK971Nh/2wWHeQviipHHcsxZtPA== 10, 24 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 ==============================End of - Headers==============================
==============================Original Headers============================== 18, 35 -- From mager-at-interchange.ubc.ca Mon May 14 10:41:03 2007 18, 35 -- Received: from mr7.mail-relay.ubc.ca (mr7.mail-relay.ubc.ca [137.82.45.13]) 18, 35 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4EFf3t3017958 18, 35 -- for {microscopy-at-microscopy.com} ; Mon, 14 May 2007 10:41:03 -0500 18, 35 -- Received: from mr7.mail-relay.ubc.ca (localhost [127.0.0.1]) 18, 35 -- by localhost (Postfix) with SMTP id 0F85C12CD8 18, 35 -- for {microscopy-at-microscopy.com} ; Mon, 14 May 2007 08:41:03 -0700 (PDT) 18, 35 -- Received: from mta1.interchange.ubc.ca (mta1.interchange.ubc.ca [142.103.145.69]) 18, 35 -- by mr7.mail-relay.ubc.ca (Postfix) with ESMTP 18, 35 -- for {microscopy-at-microscopy.com} ; Mon, 14 May 2007 08:41:02 -0700 (PDT) 18, 35 -- Received: from Mary (desk.staff.mmat.ubc.ca [137.82.16.178]) 18, 35 -- by smtp.interchange.ubc.ca 18, 35 -- (iPlanet Messaging Server 5.2 HotFix 1.21 (built Sep 8 2003)) 18, 35 -- with ESMTPS id {0JI100D7GFKEDM-at-smtp.interchange.ubc.ca} for 18, 35 -- microscopy-at-microscopy.com; Mon, 14 May 2007 08:41:02 -0700 (PDT) 18, 35 -- Date: Mon, 14 May 2007 08:40:31 -0700 18, 35 -- From: Mary Mager {mager-at-interchange.ubc.ca} 18, 35 -- Subject: RE: [Microscopy] SEM-help finding smooth, low-Z substrates 18, 35 -- In-reply-to: {200705141338.l4EDc805020316-at-ns.microscopy.com} 18, 35 -- To: rcsaic-at-sbcglobal.net 18, 35 -- Cc: "'Microscopy'" {microscopy-at-microscopy.com} 18, 35 -- Reply-to: mager-at-interchange.ubc.ca 18, 35 -- Message-id: {0JI100D7JFKEDM-at-smtp.interchange.ubc.ca} 18, 35 -- Organization: Materials Eng. UBC 18, 35 -- MIME-version: 1.0 18, 35 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 18, 35 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 18, 35 -- Content-type: text/plain; charset=us-ascii 18, 35 -- Content-transfer-encoding: 7bit 18, 35 -- Thread-index: AceWLRvzitVWDnmlQTWENpgnIDjrzAAED52g 18, 35 -- X-UBC-Scanned: Sophos PureMessage 5.3.1.294258, Antispam-Engine: 2.5.1.298604, Antispam-Data: 2007.5.14.82433 18, 35 -- X-UBC-Relayed: Relayed through mail-relay.ubc.ca 18, 35 -- X-PerlMx-Spam: Probability=7%, Report=__C230066_P5 0, __CP_URI_IN_BODY 0, __CT 0, __CTE 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __HAS_X_MAILER 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0, __STOCK_PHRASE_24 0, __USER_AGENT_MS_GENERIC 0 18, 35 -- X-Spam-Level: 18, 35 -- X-Spam-Flag: No ==============================End of - Headers==============================
Just to expand on Mary's suggestion, yes, most of the EM Supply houses sell them including us at Ladd Research. See our Specially Smooth Carbon Planchets at http://www.laddresearch.com/General_Catalog/Chapter_4/Specimen_Mounts/plan/plan.html
DISCLAIMER: Ladd Research has been selling EM supplies including carbon planchets for more than 50 years
At 11:45 AM 5/14/2007, you wrote:
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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==============================Original Headers============================== 14, 27 -- From jd-at-laddresearch.com Mon May 14 10:52:12 2007 14, 27 -- Received: from lancia.electric.net (lancia.electric.net [216.129.90.248]) 14, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4EFqBUI029566 14, 27 -- for {microscopy-at-microscopy.com} ; Mon, 14 May 2007 10:52:12 -0500 14, 27 -- Received: from root by lancia.electric.net with emc1-ok (Exim 4.62) 14, 27 -- (envelope-from {jd-at-laddresearch.com} ) 14, 27 -- id 1HncqI-0000XG-WD; Mon, 14 May 2007 08:52:11 -0700 14, 27 -- Received: by emcmailer; Mon, 14 May 2007 08:52:10 -0700 14, 27 -- Received: from [216.204.198.170] (helo=NewServer.laddresearch.com) 14, 27 -- by lancia.electric.net with esmtps (TLSv1:AES256-SHA:256) 14, 27 -- (Exim 4.62) 14, 27 -- (envelope-from {jd-at-laddresearch.com} ) 14, 27 -- id 1HncqH-0000VR-VM; Mon, 14 May 2007 08:52:10 -0700 14, 27 -- X-Mailer: QUALCOMM Windows Eudora Version 7.1.0.9 14, 27 -- Date: Mon, 14 May 2007 11:51:36 -0400 14, 27 -- To: mager-at-interchange.ubc.ca 14, 27 -- From: jd {jd-at-laddresearch.com} 14, 27 -- Subject: Re: [Microscopy] RE: SEM-help finding smooth, low-Z substrates 14, 27 -- Cc: Microscopy listserver {microscopy-at-microscopy.com} 14, 27 -- In-Reply-To: {200705141545.l4EFjNMR028507-at-ns.microscopy.com} 14, 27 -- References: {200705141545.l4EFjNMR028507-at-ns.microscopy.com} 14, 27 -- Mime-Version: 1.0 14, 27 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 14, 27 -- X-Outbound-IP: 216.204.198.170 14, 27 -- X-Env-From: jd-at-laddresearch.com 14, 27 -- X-Virus-Status: Scanned by VirusSMART (c) 14, 27 -- Message-Id: {E1HncqI-0000XG-WD-at-lancia.electric.net} ==============================End of - Headers==============================
The Delaware Biotechnology Institute (DBI) is looking to fill a part-time temporary position of microscopist in its state-of-the-art Bioimaging Center.
DBI, a unit of the University of Delaware, was established in 1999 to position Delaware as a leader in biotechnology, is a partnership involving state government, the Delaware institutions of higher education, and area industry, and is housed in a 72,000 ft2 state-of-the-art research facility located at the Delaware Technology Park in Newark, DE. DBI’s mission is to engage in leading-edge scientific discovery in the life sciences, provide biotechnology-based education, and promote economic development. The major interdisciplinary research areas include human health, agriculture, including plant molecular biology and avian genomics, environmental ecosystems, and biomaterials.
The Institute houses several core facilities including a microarray center, a bioinformatics center and a bioimaging center. The bioimaging center houses a broad range of microscopy instrumentation, including conventional fluorescence, confocal, multiphoton, atomic force, laser microdissection, transmission and field emission scanning microscopes and their ancillary sample preparation equipment. Additional details are available on the web at www.dbi.udel.edu/bioimaging.
Under the general direction of the Director of the Bioimaging Center, the part-time microscopist will provide research support for light and confocal microscopy as well as scanning and transmission electron microscopy experiments in a multi-user environment. This includes, but is not limited to training users and/or performing all steps of experimental design, sample preparation, data acquisition and analysis. This part-time temporary position is available immediately and at least through December 31, 2007. Candidates should have a minimum of a Bachelor’s degree in the life sciences. Prior microscopy experience would be highly advantageous. Compensation is commensurate with experience. Please inquire or send resume via email to Dr. Kirk Czymmek (kirk-at-udel.edu).
Kirk J. Czymmek, Ph.D. 15 Innovation Way, Suite 117 Delaware Biotechnology Institute University of Delaware Newark DE 19711 (302) 831-3450 kirk-at-udel.edu
==============================Original Headers============================== 8, 18 -- From kirk-at-udel.edu Mon May 14 11:01:55 2007 8, 18 -- Received: from md2.nss.udel.edu (md2.nss.udel.edu [128.175.1.12]) 8, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4EG1tOo008725 8, 18 -- for {Microscopy-at-microscopy.com} ; Mon, 14 May 2007 11:01:55 -0500 8, 18 -- Received: from [128.175.253.124] (host-253-124.nss.udel.edu [128.175.253.124]) 8, 18 -- by md2.nss.udel.edu (MOS 3.8.2-GA) 8, 18 -- with ESMTP id EIB03720; 8, 18 -- Mon, 14 May 2007 12:01:54 -0400 (EDT) 8, 18 -- Message-ID: {464887EB.10806-at-udel.edu} 8, 18 -- Date: Mon, 14 May 2007 12:01:47 -0400 8, 18 -- From: kirk {kirk-at-udel.edu} 8, 18 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.12) Gecko/20050915 8, 18 -- X-Accept-Language: en-us, en 8, 18 -- MIME-Version: 1.0 8, 18 -- To: Microscopy-at-microscopy.com 8, 18 -- Subject: Temporary Microscopist Position 8, 18 -- Content-Type: text/plain; charset=windows-1252; format=flowed 8, 18 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
Peter, If you thin the samples, then a perfect holding device is the adhesive on a Post-it(TM) note. If you don't thin your sample, but can scribe it well enough, then consider using a flat silicone sheet to hold the sample. Both will hold the sample for your cleave and both are just resilient enough. At 90-100 µm thick, the Post-It(TM) note works great for the MicroCleave(TM) technique. You have to shear it off the pack so that it doesn't have any curl and then using regular tape, tape it to a flat block with the adhesive side up. Carefully place your samples on it, being careful not to put the good side down on the adhesive -you won't get the adhesive off. On the rough ground down side, the adhesive doesn't stick when you lift the sample off.
-Scott
Scott D. Walck, Ph.D. Technical Director South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
US Toll Free: 1-800-728-2233 Tel: (949) 492-2600 Fax: (949) 492-1499
www.southbaytech.com walck-at-southbaytech.com
-----Original Message----- X-from: peter.tomic-at-renwireless.com [mailto:peter.tomic-at-renwireless.com] Sent: Monday, May 14, 2007 8:37 AM To: Walck-at-SouthBayTech.com
Yes, John, it's 8 mm, not microns. The issue is holding the device while making the second cleave or cut. The sample would have to be mounted on something to use a diamond saw, which is available. However, I do need to keep the face of the surface I'm looking at undisturbed with whatever method I use.
Regards,
Peter
Mardinly, John wrote: } ~ 8 mm apart? Did I read this correctly? Not microns? We use a Disco } saw routinely. If it's ~ 8 mm apart, you can use any diamond saw. If } it's microns you need a FIB. } } John Mardinly } Intel Corporation } This email does not represent an opinion of Intel Corporation. } } -----Original Message----- } From: peter.tomic-at-renwireless.com [mailto:peter.tomic-at-renwireless.com] } Sent: Tuesday, May 08, 2007 10:33 AM } To: Mardinly, John } Subject: [Microscopy] Silicon Cross-section sample preparation } } } } } ---------------------------------------------------------------------- } -- } ---- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -- } ---- } } Microscopy Folks, } } I'd like to get some input on sample preparation with respect to } silicon. } } PROBLEM: } } I have an FEI XL-50 FESEM that was originally designed to accept flat } samples, essentially silicon wafers. It does not have what one would } consider a typical exchange port. The exchange port is robotically } controlled, and the maximum sample height, including stub, must be 5 } mm or less. I must view these samples on edge, i.e. 90 degrees. This } presents a problem in that I must cleave two parallel sections very } close to each other. Perhaps diamond cutters can do this, but my } hands are too shaky. } } QUESTION: } } Is there a device, or method, that would allow me to make these } cleaves ~ 8 mm apart with any reasonable control? I found stubs that } are low profile, 38 and 90 degrees, from the nice people at Ted Pella, } but I still have this issue of doing two parallel cleaves very close together. } } Just for info. purposes I am in the silicon MEMS development arena. } } If you feel your reply is of general interest to this community, } please reply to all, or you may contact me directly. } } Regards to all in this small world, } } Peter Tomic } Renaissance Wireless Corp. } } }
==============================Original Headers============================== 6, 26 -- From peter.tomic-at-renwireless.com Mon May 14 10:33:20 2007 6, 26 -- Received: from mail.rw.local (4.piok2.xdsl.nauticom.net [209.195.176.101]) 6, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4EFXJSP006437 6, 26 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 14 May 2007 10:33:20 -0500 6, 26 -- Received: from localhost (localhost [127.0.0.1]) 6, 26 -- by mail.rw.local (Postfix) with ESMTP id D2F46246B5; 6, 26 -- Mon, 14 May 2007 11:33:17 -0400 (EDT) 6, 26 -- Received: from mail.rw.local ([127.0.0.1]) 6, 26 -- by localhost (mail.rw.local [127.0.0.1]) (amavisd-new, port 10024) with ESMTP 6, 26 -- id 26580-04; Mon, 14 May 2007 11:33:17 -0400 (EDT) 6, 26 -- Received: from [127.0.0.1] (unknown [10.11.10.110]) 6, 26 -- by mail.rw.local (Postfix) with ESMTP id A2C0918AC7; 6, 26 -- Mon, 14 May 2007 11:33:16 -0400 (EDT) 6, 26 -- Message-ID: {464882B3.7010905-at-renwireless.com} 6, 26 -- Date: Mon, 14 May 2007 11:39:31 -0400 6, 26 -- From: Peter Tomic {peter.tomic-at-renwireless.com} 6, 26 -- User-Agent: Thunderbird 2.0.0.0 (Windows/20070326) 6, 26 -- MIME-Version: 1.0 6, 26 -- To: "Mardinly, John" {john.mardinly-at-intel.com} 6, 26 -- CC: Microscopy-at-MSA.Microscopy.Com 6, 26 -- Subject: Re: [Microscopy] Silicon Cross-section sample preparation 6, 26 -- References: {F3CB8931ABF8294DB889E977150CAD68705D13-at-scsmsx415.amr.corp.intel.com} 6, 26 -- In-Reply-To: {F3CB8931ABF8294DB889E977150CAD68705D13-at-scsmsx415.amr.corp.intel.com} 6, 26 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 6, 26 -- Content-Transfer-Encoding: 7bit 6, 26 -- X-Virus-Scanned: amavisd-new at renwireless.com ==============================End of - Headers==============================
==============================Original Headers============================== 16, 23 -- From walck-at-southbaytech.com Mon May 14 13:07:43 2007 16, 23 -- Received: from flpvm09.prodigy.net (flpvm09.prodigy.net [207.115.20.39]) 16, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4EI7hRk022667 16, 23 -- for {Microscopy-at-microscopy.com} ; Mon, 14 May 2007 13:07:43 -0500 16, 23 -- X-ORBL: [64.169.217.123] 16, 23 -- Received: from dynamicbl8uno3 (adsl-64-169-217-123.dsl.lsan03.pacbell.net [64.169.217.123]) 16, 23 -- by flpvm09.prodigy.net (8.13.8 out.dk.spool/8.13.8) with ESMTP id l4EI7df1007427; 16, 23 -- Mon, 14 May 2007 11:07:41 -0700 16, 23 -- From: "Scott Walck" {walck-at-southbaytech.com} 16, 23 -- To: {peter.tomic-at-renwireless.com} 16, 23 -- Cc: {Microscopy-at-microscopy.com} 16, 23 -- Subject: RE: [Microscopy] Re: Silicon Cross-section sample preparation 16, 23 -- Date: Mon, 14 May 2007 11:07:58 -0700 16, 23 -- Message-ID: {010201c79652$ce186b60$7801a8c0-at-dynamicbl8uno3} 16, 23 -- MIME-Version: 1.0 16, 23 -- Content-Type: text/plain; 16, 23 -- charset="iso-8859-1" 16, 23 -- X-Mailer: Microsoft Office Outlook 11 16, 23 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 16, 23 -- In-Reply-To: {200705141536.l4EFaj78012214-at-ns.microscopy.com} 16, 23 -- Thread-Index: AceWPa4D2Gu39IP8TVq1FJNzIDTikAAFGIyw 16, 23 -- Content-Transfer-Encoding: 8bit 16, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4EI7hRk022667 ==============================End of - Headers==============================
On May 14, 2007, at 6:29 AM, rcsaic-at-sbcglobal.net wrote:
} A substrate for dispersing the particles that would fit two } requirements: 1) have a pretty low average Z (carbon would be ideal } but I'm } willing to consider other ideas), so that my particles will stand out } strongly in BSE, and then 2) be as smooth as possible on the sub-micron } scale. This last requirement is the tricky one because although there } are } lots of carbon substrates available commercially, so far we haven't } found } any that don't have sub-micron scratches and other roughness such that } particles in the 0.1 micron size range don't get lost in the surface } topography
Dear Roy, Have you tried carbon evaporated onto freshly-cleaved mica? The layer can be floated off and deposited onto lacy carbon so that there will be many areas several um across that have only the thin carbon substrate. The layer can be made to be only tens of nm thick, so it is very unlikely to have ~100 nm steps. Yours, Bill Tivol, PhD EM Scientist Electron Cryo-Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
==============================Original Headers============================== 5, 22 -- From tivol-at-caltech.edu Mon May 14 13:14:08 2007 5, 22 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 5, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4EIE7be030887 5, 22 -- for {microscopy-at-msa.microscopy.com} ; Mon, 14 May 2007 13:14:07 -0500 5, 22 -- Received: from water-dog.its.caltech.edu (water-dog [192.168.1.26]) 5, 22 -- by earth-ox-postvirus (Postfix) with ESMTP id 935D91AB53 5, 22 -- for {microscopy-at-msa.microscopy.com} ; Mon, 14 May 2007 11:14:05 -0700 (PDT) 5, 22 -- Received: from [192.168.159.233] (jpix-01.caltech.edu [131.215.2.133]) 5, 22 -- by water-ox.its.caltech.edu (Postfix) with ESMTP id 308CF1AB75 5, 22 -- for {microscopy-at-msa.microscopy.com} ; Mon, 14 May 2007 11:13:58 -0700 (PDT) 5, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 5, 22 -- In-Reply-To: {200705141329.l4EDTTaf011644-at-ns.microscopy.com} 5, 22 -- References: {200705141329.l4EDTTaf011644-at-ns.microscopy.com} 5, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 5, 22 -- Message-Id: {6cf514fb3021c0813cf36c66e1b77213-at-caltech.edu} 5, 22 -- Content-Transfer-Encoding: 7bit 5, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 5, 22 -- Subject: Re: [Microscopy] SEM-help finding smooth, low-Z substrates 5, 22 -- Date: Mon, 14 May 2007 11:25:40 -0700 5, 22 -- To: microscopy-at-msa.microscopy.com 5, 22 -- X-Mailer: Apple Mail (2.624) 5, 22 -- X-Spam-Scanned: at Caltech-ITS on water-dog by amavisd-2.4.5 ==============================End of - Headers==============================
We have a Leica EMPACT high pressure freezer and upon last start up the hydraulic fluid reservoir had gone dry. I refilled with MCH and followed the operation manual's instructions on getting air bubbles out of the line (basically just using a long skinny rod to plunge the sealing "bb" at the bottom of the reservoir until the bubbles stop coming out). Now I'm getting an error message that prompts me to fill the chamber with hydraulic fluid.
I can reboot the system and get rid of the message, but only for one freezing run---then it returns. Many reboots, freezes, and lots of bb plunging later, I'm still getting the message. The pressure readings during the HPF runs have been mostly good, but there is an occasional stinker----more than I remember on average than in past runs.
I'm guessing there's an elusive bubble in the line somewhere, but has anyone else run across this? More to the point, does anyone have an easy fix?
Thanks all.
Randy
Randy Tindall Senior EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount= Week&NavType=Both&Type=TimePlan
==============================Original Headers============================== 8, 23 -- From TindallR-at-missouri.edu Mon May 14 13:26:59 2007 8, 23 -- Received: from um-tsmtpout1.um.umsystem.edu (um-tsmtpout1.um.umsystem.edu [209.106.228.23]) 8, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4EIQweR013489 8, 23 -- for {microscopy-at-microscopy.com} ; Mon, 14 May 2007 13:26:58 -0500 8, 23 -- Received: from UM-XMAIL08.um.umsystem.edu ([209.106.228.34]) by um-tsmtpout1.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 8, 23 -- Mon, 14 May 2007 13:26:58 -0500 8, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 8, 23 -- Content-class: urn:content-classes:message 8, 23 -- MIME-Version: 1.0 8, 23 -- Content-Type: text/plain; 8, 23 -- charset="us-ascii" 8, 23 -- Subject: Leica HPF EMPACT question 8, 23 -- Date: Mon, 14 May 2007 13:26:58 -0500 8, 23 -- Message-ID: {91108EF9255B394CBF8B7E3789814A4101C0B9F5-at-UM-XMAIL08.um.umsystem.edu} 8, 23 -- X-MS-Has-Attach: 8, 23 -- X-MS-TNEF-Correlator: 8, 23 -- Thread-Topic: Leica HPF EMPACT question 8, 23 -- Thread-Index: AceWVXTtFM4T49y8Tvi3hVD3DhO5/A== 8, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 8, 23 -- To: {microscopy-at-microscopy.com} 8, 23 -- X-OriginalArrivalTime: 14 May 2007 18:26:58.0845 (UTC) FILETIME=[755834D0:01C79655] 8, 23 -- Content-Transfer-Encoding: 8bit 8, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4EIQweR013489 ==============================End of - Headers==============================
The Zatkoff (www.zatkoff.com) company also carries an extensive stock of O-rings. Before I tried making an O-ring on my own I would certainly check with them to see if they couldn't furnish something to meet my needs. -- Wilbur C. Bigelow, Professor Emeritus Materials Sci. & Engr., Univ. of Michigan Ann Arbor, Michigan 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-975-0858 Address mail to: 2911 Whittier Court Ann Arbor, MI 48104-6731
==============================Original Headers============================== 1, 14 -- From bigelow-at-umich.edu Mon May 14 16:23:01 2007 1, 14 -- Received: from hackers.mr.itd.umich.edu (smtp.mail.umich.edu [141.211.14.81]) 1, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4ELN0WN027922 1, 14 -- for {microscopy-at-microscopy.com} ; Mon, 14 May 2007 16:23:00 -0500 1, 14 -- Received: FROM [141.212.131.221] (bigelow-g4.engin.umich.edu [141.212.131.221]) 1, 14 -- BY hackers.mr.itd.umich.edu ID 4648D334.113A0.21325 ; 1, 14 -- 14 May 2007 17:23:00 -0400 1, 14 -- Mime-Version: 1.0 1, 14 -- Message-Id: {p06210201c26e82f8bc7e-at-[141.212.131.221]} 1, 14 -- Date: Mon, 14 May 2007 17:22:58 -0400 1, 14 -- To: Microscopy Listserver {microscopy-at-microscopy.com} 1, 14 -- From: Wil Bigelow {bigelow-at-umich.edu} 1, 14 -- Subject: [Microscopy] R: O-rings 1, 14 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
Roy Christoffersen wrote: ==============================================================
We could use some suggestions/collective experience about solid substrates for doing SEM imaging of dispersed small particles.
What we are imaging: silicate and oxide mineral grains and silicate glass particles down to the 0.1 micron size range (maybe smaller). Objective is to disperse the particle on a substrate and do total particle counting and automated measuring without "losing" the smallest particles from the data set.
What we need: A substrate for dispersing the particles that would fit two requirements: 1) have a pretty low average Z (carbon would be ideal but I'm willing to consider other ideas), so that my particles will stand out strongly in BSE, and then 2) be as smooth as possible on the sub-micron scale. This last requirement is the tricky one because although there are lots of carbon substrates available commercially, so far we haven't found any that don't have sub-micron scratches and other roughness such that particles in the 0.1 micron size range don't get lost in the surface topography.
Well, that's about all I can think of. Thanks to all of you in advance.
Roy Christoffersen FE-STEM Facility Manager ARES Directorate NASA Johnson Space Center, Houston, TX rcsaic-at-sbcglobal.net ======================================================================= In my opinion, you should consider HOPG (highly ordered pyrolytic graphite), see URL
For those who are not familiar with this unique material and its novel properties, it can be cleaved much like mica into very thin strippings, actually more easily than mica. This is done by pressing a piece of Scotch tape, either single sided or double sided onto the flat plate surface of the HOPG and then literally, stripping off a layer of HOPG which is left strongly adhering to the tape. This tape can then be mounted on a conventional SEM mount, perhaps with a double sided conductive carbon disc. Many additional strippings can be made from the resulting block until it is all consumed.
The resulting stripping is highly (mirror) reflective, demonstrating the virtually zero porosity in the HOPG stripped layer. And depending on the grade of HOPG selected, there will be regions of varying area sizes of atomically smooth HOPG. You will not "lose" particles in the structure of the HOPG.
There have been some suggestions by others, but even though we too offer glassy carbon, we recommend the HOPG over the glassy carbon for several reasons: a) Glassy carbon exhibits some porosity (depending on grade) on the scale of your fine particles that you don't want to lose and b) since one can get a number of individual strippings out of each HOPG plate, the cost per stripping (e.g. sample) is far less than the cost of a piece of glassy carbon.
HOPG is used widely in the field of AFM, one of the reasons being exactly the reason you gave: ".......don't have sub-micron scratches and other roughness such that particles in the 0.1 micron size range don't get lost in the surface topography". Your specifications are consistent with the needs of AFM users looking at nano-sized particles (which can not be cleaved).
Disclaimer: SPI Supplies offers a complete line of HOPG and glassy carbon products and therefore we have a vested interest in seeing more of these items being used by researchers. See URL http://www.2spi.com/catalog/mounts/vitreous.html
Chuck
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==============================Original Headers============================== 21, 20 -- From cgarber-at-2spi.com Mon May 14 16:47:47 2007 21, 20 -- Received: from diskless5.axs2000.net (mail.axs2000.net [209.120.196.43]) 21, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4ELllMw007235 21, 20 -- for {microscopy-at-msa.microscopy.com} ; Mon, 14 May 2007 16:47:47 -0500 21, 20 -- Received: from [192.168.122.43] ([212.150.94.34]) 21, 20 -- (authenticated bits=0) 21, 20 -- by diskless5.axs2000.net (8.12.11/8.12.11) with ESMTP id l4ELli5J018723; 21, 20 -- Mon, 14 May 2007 17:47:45 -0400 21, 20 -- X-IDV-FirstRcvd: [212.150.94.34] 21, 20 -- X-IDV-HELO: [192.168.122.43] 21, 20 -- X-IDV-Authenticated-User: cgarber 21, 20 -- Message-ID: {4648D8D9.8050209-at-2spi.com} 21, 20 -- Date: Mon, 14 May 2007 17:47:05 -0400 21, 20 -- From: "Garber, Charles A" {cgarber-at-2spi.com} 21, 20 -- User-Agent: Thunderbird 2.0.0.0 (Windows/20070326) 21, 20 -- MIME-Version: 1.0 21, 20 -- To: Microscopy Listserver {microscopy-at-msa.microscopy.com} 21, 20 -- Subject: SEM-help finding smooth, low-Z substrates 21, 20 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 21, 20 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
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Question: Roy, Although I normall don't work with particles that small, most of our particle analysis is performed on carbon conductive tabs from Ted Pella Inc (catalog # 16084-4). There is some texture to the tabs, but usually the particles stand proud of the tab. With a backscatter image, the higher z particles will stand out from the carbon. The problem with analyzing such a fine powder will be particle separation. We tend to sprinkle a small sample on the tab instead of pressing the tab into the powder, just to get a dispersion, provide spearation between particles, and to have the particle be proud of the tab.
Gerald Shulke Materials Characterization Labs DaimlerChrysler- Chrysler Group
Looks like Carbon is the idea material for your research. Have you tried to use natural graphite crystal -- it has beautiful layer structure and large flat surface area as well. Find a geologist and ask a piece of graphic crystal, stick it onto your SEM stub, then peel off first 3-10 layers to have fresh surface ready for use. Use sharp knife and peel from edge -- I saw some other people used tape to peel the top layers off. We used to make fresh "flaw-free" graphite surface for STM analysis.
Regards, ---------------------------------------------- Xiang Yang Electron Microscopy Center Saint Mary's University Science Building, Suite 135 Halifax, NS Canada B3H 3C3 Tel: (902) 496-8292 (lab) (902) 420-5709 (off) http://fgsr.smu.ca/emc/
----- Original Message ----- X-from: {rcsaic-at-sbcglobal.net} To: {xyang-at-SMU.CA} Sent: Monday, May 14, 2007 10:34 AM
Roy,
The artful use of a heat gun directed at a hot melt glue covered SEM stub will produce a lustrous surface on the low Z, low outgassing glue substrate. Perhaps while the glue is still warm and adhesive, you could sprinkle your sample onto it.
Bart Cannon Cannon Microprobe Seattle
==============================Original Headers============================== 4, 22 -- From cannonmp-at-comcast.net Mon May 14 19:50:45 2007 4, 22 -- Received: from rwcrmhc14.comcast.net (rwcrmhc14.comcast.net [204.127.192.84]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4F0oi49018028 4, 22 -- for {Microscopy-at-microscopy.com} ; Mon, 14 May 2007 19:50:45 -0500 4, 22 -- Received: from bartg84ti61htn (c-71-197-166-133.hsd1.or.comcast.net[71.197.166.133]) 4, 22 -- by comcast.net (rwcrmhc14) with SMTP 4, 22 -- id {20070515005044m1400iou33e} ; Tue, 15 May 2007 00:50:44 +0000 4, 22 -- Message-ID: {003601c7968a$cced1050$85a6c547-at-bartg84ti61htn} 4, 22 -- From: "cannonmp" {cannonmp-at-comcast.net} 4, 22 -- To: {Microscopy-at-microscopy.com} 4, 22 -- Subject: Smooth Low Z Substrate 4, 22 -- Date: Mon, 14 May 2007 17:48:48 -0700 4, 22 -- MIME-Version: 1.0 4, 22 -- Content-Type: text/plain; 4, 22 -- format=flowed; 4, 22 -- charset="iso-8859-1"; 4, 22 -- reply-type=original 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- X-Priority: 3 4, 22 -- X-MSMail-Priority: Normal 4, 22 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2180 4, 22 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 ==============================End of - Headers==============================
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Email: tnkeim-at-sbcglobal.net Name: Thomas Keim
Organization: Dentist
Title-Subject: [Filtered] Run Zeiss 100W from Instek Power Supply
Question: Has anyone tried powering the Zeiss 100W halogen light source (Zeiss 467259, for Standards and Photomicroscopes) with an Intek GPR-1810HD power supply? Any issues?
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Title-Subject: [Filtered] Cleaning Surfaces of Polysulfone membrane for SEM
Question: Hi,
I am doing SEM and FESEM scanning of Polysulfone membrane. Although I always wash the membrane with water and alchol prior to scanning but I am getting foreign particles on new (comes sealed from industry) and casted membranes surface. I dont know what these particle are how to clean them ( I suppose they are some dust embedded on porous membrane surface)
Can anybody tell me some standard process to get rid of these foreign particles on the surface. I can send Pics showing these particles on the surface.
I have obtained a sample of freeze-dried bacteria, and would like to prepare it for the SEM. Is there something more than just double-stick carbon disk on a stub and sputter coating if it's already freeze dried?
--Justin A. Kraft Atlantis Academy
==============================Original Headers============================== 2, 26 -- From kraftpiano-at-gmail.com Tue May 15 07:33:16 2007 2, 26 -- Received: from nz-out-0506.google.com (nz-out-0506.google.com [64.233.162.233]) 2, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4FCXGpg014226 2, 26 -- for {microscopy-at-microscopy.com} ; Tue, 15 May 2007 07:33:16 -0500 2, 26 -- Received: by nz-out-0506.google.com with SMTP id x3so124604nzd 2, 26 -- for {microscopy-at-microscopy.com} ; Tue, 15 May 2007 05:33:16 -0700 (PDT) 2, 26 -- DKIM-Signature: a=rsa-sha1; c=relaxed/relaxed; 2, 26 -- d=gmail.com; s=beta; 2, 26 -- h=domainkey-signature:received:received:message-id:date:from:to:subject:mime-version:content-type:content-transfer-encoding:content-disposition; 2, 26 -- b=lgLXtNSv/bWorA7IhC3jUw/KVSe9uu7bmLhxbt2V9xRrQCXBJMG+GhN5dhyy5KVrbgWIQF9gYIRigCR6Pz8jXcq5ZeOTMfUSL/UJwmiE4V1xXlm4smpxGReB08HBVcN4EKlC/wDd9I9Vq2q6JfRDE8K9ElZ9DZhNSvL87ivdU8E= 2, 26 -- DomainKey-Signature: a=rsa-sha1; c=nofws; 2, 26 -- d=gmail.com; s=beta; 2, 26 -- h=received:message-id:date:from:to:subject:mime-version:content-type:content-transfer-encoding:content-disposition; 2, 26 -- b=TWJE/WPCtkZ9cdZAQ63AdcRXNVqxnniuEL7zlKHyNBfvgXEf3w3ekcjJdbJEA/7Hw93QMAfOWzhFtTzULPLjSf3Tm89tyVe58nH7VinyqWnYLoujWEMva9D2rLepbNDM4ZbJZUZdsLTHC/lpyGVSgCIpdky7swJKV3mRnoyJ/h4= 2, 26 -- Received: by 10.114.149.2 with SMTP id w2mr643190wad.1179232395835; 2, 26 -- Tue, 15 May 2007 05:33:15 -0700 (PDT) 2, 26 -- Received: by 10.114.78.15 with HTTP; Tue, 15 May 2007 05:33:15 -0700 (PDT) 2, 26 -- Message-ID: {25e2b0d20705150533t7a830a14w4fd0094468659a64-at-mail.gmail.com} 2, 26 -- Date: Tue, 15 May 2007 08:33:15 -0400 2, 26 -- From: "Justin Kraft" {kraftpiano-at-gmail.com} 2, 26 -- To: microscopy-at-microscopy.com 2, 26 -- Subject: SEM Bacterial sample prep. 2, 26 -- MIME-Version: 1.0 2, 26 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 2, 26 -- Content-Transfer-Encoding: 7bit 2, 26 -- Content-Disposition: inline ==============================End of - Headers==============================
I'd like to get some high-magnification images of the 1.00mm tungsten carbide balls that we use in our ball point and roller ball pens. The images I've been able to get with light microscopy do not show enough surface detail. Can anyone recommend the best type of imaging for this study and a facility that will do that sort of imaging for us inexpensively?
Thanks very much,
Robert Zonis Product Development Chemist, LMTC Sanford L.P. - A Newell Rubbermaid Company 3 Sharpie Way Shelbyville, TN 37160 Direct: +1 (931) 685-6635 Fax: +1 (931) 685-6613 robert.zonis-at-sanford.com www.papermate.com
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==============================Original Headers============================== 9, 34 -- From Robert.Zonis-at-sanford.com Tue May 15 08:08:47 2007 9, 34 -- Received: from mail51.messagelabs.com (mail51.messagelabs.com [216.82.244.51]) 9, 34 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l4FD8kf9003197 9, 34 -- for {Microscopy-at-microscopy.com} ; Tue, 15 May 2007 08:08:47 -0500 9, 34 -- X-VirusChecked: Checked 9, 34 -- X-Env-Sender: Robert.Zonis-at-sanford.com 9, 34 -- X-Msg-Ref: server-11.tower-51.messagelabs.com!1179234524!43756649!1 9, 34 -- X-StarScan-Version: 5.5.10.7.1; banners=sanford.com,-,- 9, 34 -- X-Originating-IP: [12.2.115.11] 9, 34 -- Received: (qmail 17956 invoked from network); 15 May 2007 13:08:44 -0000 9, 34 -- Received: from gnat.newellco.com (HELO nafepncsvpout2.nr.ad.newellco.com) (12.2.115.11) 9, 34 -- by server-11.tower-51.messagelabs.com with SMTP; 15 May 2007 13:08:44 -0000 9, 34 -- Received: from nafepncsefe02.nr.ad.newellco.com (unverified) by nafepncsvpout2.nr.ad.newellco.com 9, 34 -- (Content Technologies SMTPRS 4.3.17) with ESMTP id {T7f9b30f9650a059a2116e4-at-nafepncsvpout2.nr.ad.newellco.com} for {Microscopy-at-microscopy.com} ; 9, 34 -- Tue, 15 May 2007 08:08:44 -0500 9, 34 -- Received: from nashbsasebe01.nr.ad.newellco.com ([10.217.223.198]) by nafepncsefe02.nr.ad.newellco.com with Microsoft SMTPSVC(6.0.3790.1830); 9, 34 -- Tue, 15 May 2007 08:08:44 -0500 9, 34 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 9, 34 -- Content-class: urn:content-classes:message 9, 34 -- MIME-Version: 1.0 9, 34 -- Content-Type: text/plain; 9, 34 -- charset="iso-8859-1" 9, 34 -- Subject: High-magnification imaging help and recommendations 9, 34 -- Date: Tue, 15 May 2007 08:08:41 -0500 9, 34 -- Message-ID: {8809C39726DC5F43824A3D3BE5D0286A0C868313-at-nashbsasebe01.nr.ad.newellco.com} 9, 34 -- X-MS-Has-Attach: 9, 34 -- X-MS-TNEF-Correlator: 9, 34 -- Thread-Topic: High-magnification imaging help and recommendations 9, 34 -- Thread-Index: AceW8b9UX12A6aMCTM2NqlyFcBlw0QAADgrg 9, 34 -- From: "Zonis, Robert" {Robert.Zonis-at-sanford.com} 9, 34 -- To: {Microscopy-at-microscopy.com} 9, 34 -- X-OriginalArrivalTime: 15 May 2007 13:08:44.0328 (UTC) FILETIME=[2A89E280:01C796F2] 9, 34 -- Content-Transfer-Encoding: 8bit 9, 34 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4FD8kf9003197 ==============================End of - Headers==============================
Hi Randy, Sounds like a problem we had recently with our EM PACT2. We tried draining the entire hydraulic fluid chamber and refilling, as well as using a stick to release any air bubbles. This was still not working, and the problem ended up being a failed detector for the fluid level. We contacted Leica and received a new detector for the instrument, which was quite easy to install. So you may want to contact them as well to discuss the problem. Hope that helps!
Shauna
Shauna deVarennes, B.Sc, AHT, RLAT Research Technician Diagnostic Imaging and Microscopy Viral Diseases Division National Microbiology Laboratory
Canadian Science Centre for Human and Animal Health 1015 Arlington Street, Room H4125 Winnipeg, Manitoba, R3E 3R2
We have a Leica EMPACT high pressure freezer and upon last start up the hydraulic fluid reservoir had gone dry. I refilled with MCH and followed the operation manual's instructions on getting air bubbles out of the line (basically just using a long skinny rod to plunge the sealing "bb" at the bottom of the reservoir until the bubbles stop coming out). Now I'm getting an error message that prompts me to fill the chamber with hydraulic fluid.
I can reboot the system and get rid of the message, but only for one freezing run---then it returns. Many reboots, freezes, and lots of bb plunging later, I'm still getting the message. The pressure readings during the HPF runs have been mostly good, but there is an occasional stinker----more than I remember on average than in past runs.
I'm guessing there's an elusive bubble in the line somewhere, but has anyone else run across this? More to the point, does anyone have an easy fix?
Thanks all.
Randy
Randy Tindall Senior EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
==============================Original Headers============================== 13, 20 -- From shauna_devarennes-at-phac-aspc.gc.ca Tue May 15 09:38:33 2007 13, 20 -- Received: from scmze002.ssan.egs-seg.gc.ca (scmze002.ssan.egs-seg.gc.ca [205.194.19.86]) 13, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4FEcWhD016973 13, 20 -- for {Microscopy-at-microscopy.com} ; Tue, 15 May 2007 09:38:32 -0500 13, 20 -- X-SBRS: 3.5 13, 20 -- Received: from unknown (HELO nav99.hc-sc.gc.ca) ([198.103.172.247]) 13, 20 -- by scmze000.ssan.egs-seg.gc.ca with SMTP; 15 May 2007 14:37:32 +0000 13, 20 -- Received: from smta00.hc-sc.gc.ca ([10.241.40.152]) 13, 20 -- by nav99.hc-sc.gc.ca (SMSSMTP 4.1.0.19) with SMTP id M2007051510331203974 13, 20 -- for {Microscopy-at-Microscopy.com} ; Tue, 15 May 2007 10:33:12 -0400 13, 20 -- Subject: Re: Leica HPF EM PACT question 13, 20 -- To: Microscopy-at-microscopy.com 13, 20 -- X-Mailer: Lotus Notes Release 5.0.5 September 22, 2000 13, 20 -- Message-ID: {OF871DB19C.B25E3E9E-ON862572DC.004F7E9A-at-hc-sc.gc.ca} 13, 20 -- From: Shauna deVarennes {shauna_devarennes-at-phac-aspc.gc.ca} 13, 20 -- Date: Tue, 15 May 2007 09:36:58 -0500 13, 20 -- X-MIMETrack: Serialize by Router on SMTA00/HC-SC/GC/CA(Release 6.5.5|November 30, 2005) at 13, 20 -- 2007-05-15 10:37:32 AM 13, 20 -- MIME-Version: 1.0 13, 20 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
} I have obtained a sample of freeze-dried bacteria, and would like to } prepare it for the SEM. Is there something more than just } double-stick carbon disk on a stub and sputter coating if it's } already } freeze dried? } } --Justin A. Kraft } Atlantis Academy
X-from my - limited - experience with preparing Bacteria for SEM, I can say that I would NOT recommend to just apply 'freeze-dried bacteria' to a double-stick carbon disk, and then sputter-coat the stuff. These freeze-dried bacteria are most likely completely collapsed and therefore will look 'collapsed' and 'dead'. I would suggest to recultivate the cells in a suitable culture medium, then either fix them chemically, apply them to a filter (like 'Nuclepore'), dehydrate, followed by critical-point drying and sputter coating, OR: apply them to a filter, fix them physically (plunge-freezing), then dehydrate at low temperature by freeze-drying at -80 C, then sputter-coating at low temperature or even rotary-shadowing at low-temperature ... and then SEM. Depending on the type of bacteria, many variations of these protocols are possible and might be necessary.
regards Reinhard Rachel
---------------------- PD Dr. Reinhard Rachel Universitaet Regensburg Centre for EM - NWF III - Lehrstuhl fuer Anatomie Universitaetsstr. 31 D-93053 Regensburg - Germany
==============================Original Headers============================== 8, 25 -- From reinhard.rachel-at-biologie.uni-regensburg.de Tue May 15 12:47:30 2007 8, 25 -- Received: from rrzmta2.rz.uni-regensburg.de (rrzmta2.rz.uni-regensburg.de [194.94.155.53]) 8, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4FHlUKI000390 8, 25 -- for {Microscopy-at-microscopy.com} ; Tue, 15 May 2007 12:47:30 -0500 8, 25 -- Received: from rrzmta2.rz.uni-regensburg.de (localhost [127.0.0.1]) 8, 25 -- by localhost (Postfix) with SMTP id 068594F3D; 8, 25 -- Tue, 15 May 2007 19:47:37 +0200 (CEST) 8, 25 -- Received: from localhost (donald1.rz.uni-regensburg.de [132.199.4.91]) 8, 25 -- by rrzmta2.rz.uni-regensburg.de (Postfix) with ESMTP id E90964E25; 8, 25 -- Tue, 15 May 2007 19:47:36 +0200 (CEST) 8, 25 -- Received: from pc55154.vkl.uni-regensburg.de (pc55154.vkl.uni-regensburg.de [132.199.82.169]) 8, 25 -- by webmail.uni-regensburg.de (IMP) with HTTP 8, 25 -- for {rar04520-at-rrzlic2.uni-regensburg.de} ; Tue, 15 May 2007 19:47:28 +0200 8, 25 -- Message-ID: {1179251248.4649f230cb74d-at-webmail.uni-regensburg.de} 8, 25 -- Date: Tue, 15 May 2007 19:47:28 +0200 8, 25 -- From: reinhard rachel {reinhard.rachel-at-biologie.uni-regensburg.de} 8, 25 -- To: kraftpiano-at-gmail.com, Microscopy-at-microscopy.com 8, 25 -- Subject: [Microscopy] SEM Bacterial sample prep. 8, 25 -- References: {200705151233.l4FCXUdC014990-at-ns.microscopy.com} 8, 25 -- In-Reply-To: {200705151233.l4FCXUdC014990-at-ns.microscopy.com} 8, 25 -- MIME-Version: 1.0 8, 25 -- Content-Type: text/plain; charset=ISO-8859-1 8, 25 -- Content-Transfer-Encoding: 8bit 8, 25 -- User-Agent: Internet Messaging Program (IMP) 3.2.8 8, 25 -- X-Originating-IP: 132.199.82.169 ==============================End of - Headers==============================
Dear Robert, No one else has answered your question, so I will try. I think your best bet would be to image the surface of your balls by Scanning Electron Microscopy (SEM). This should be available in the closest university or at most failure analysis consulting engineering firms. SEMs are fairly common and your requirements are fairly simple. The SEM should be able to go from 50 times magnification to 10,000 times with a good depth of focus, so you can see the details on the rounded surface. You should be able to get some good images in less than an hour. Good luck.
-----Original Message----- X-from: Robert.Zonis-at-sanford.com [mailto:Robert.Zonis-at-sanford.com] Sent: May 15, 2007 6:13 AM To: mager-at-interchange.ubc.ca
To all:
I'd like to get some high-magnification images of the 1.00mm tungsten carbide balls that we use in our ball point and roller ball pens. The images I've been able to get with light microscopy do not show enough surface detail. Can anyone recommend the best type of imaging for this study and a facility that will do that sort of imaging for us inexpensively?
Thanks very much,
Robert Zonis Product Development Chemist, LMTC Sanford L.P. - A Newell Rubbermaid Company 3 Sharpie Way Shelbyville, TN 37160 Direct: +1 (931) 685-6635 Fax: +1 (931) 685-6613 robert.zonis-at-sanford.com www.papermate.com
This message may contain information that is confidential and/or protected by law. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution, copying or communication of this message is strictly prohibited. If you have received this communication in error, please contact the sender immediately and delete the message. Please note that although we will take all commercially reasonable efforts to prevent viruses from being transmitted from our systems, it is the responsibility of the recipient to check for and prevent adverse action by viruses on its own systems.
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==============================Original Headers============================== 9, 34 -- From Robert.Zonis-at-sanford.com Tue May 15 08:08:47 2007 9, 34 -- Received: from mail51.messagelabs.com (mail51.messagelabs.com [216.82.244.51]) 9, 34 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l4FD8kf9003197 9, 34 -- for {Microscopy-at-microscopy.com} ; Tue, 15 May 2007 08:08:47 -0500 9, 34 -- X-VirusChecked: Checked 9, 34 -- X-Env-Sender: Robert.Zonis-at-sanford.com 9, 34 -- X-Msg-Ref: server-11.tower-51.messagelabs.com!1179234524!43756649!1 9, 34 -- X-StarScan-Version: 5.5.10.7.1; banners=sanford.com,-,- 9, 34 -- X-Originating-IP: [12.2.115.11] 9, 34 -- Received: (qmail 17956 invoked from network); 15 May 2007 13:08:44 -0000 9, 34 -- Received: from gnat.newellco.com (HELO nafepncsvpout2.nr.ad.newellco.com) (12.2.115.11) 9, 34 -- by server-11.tower-51.messagelabs.com with SMTP; 15 May 2007 13:08:44 -0000 9, 34 -- Received: from nafepncsefe02.nr.ad.newellco.com (unverified) by nafepncsvpout2.nr.ad.newellco.com 9, 34 -- (Content Technologies SMTPRS 4.3.17) with ESMTP id {T7f9b30f9650a059a2116e4-at-nafepncsvpout2.nr.ad.newellco.com} for {Microscopy-at-microscopy.com} ; 9, 34 -- Tue, 15 May 2007 08:08:44 -0500 9, 34 -- Received: from nashbsasebe01.nr.ad.newellco.com ([10.217.223.198]) by nafepncsefe02.nr.ad.newellco.com with Microsoft SMTPSVC(6.0.3790.1830); 9, 34 -- Tue, 15 May 2007 08:08:44 -0500 9, 34 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 9, 34 -- Content-class: urn:content-classes:message 9, 34 -- MIME-Version: 1.0 9, 34 -- Content-Type: text/plain; 9, 34 -- charset="iso-8859-1" 9, 34 -- Subject: High-magnification imaging help and recommendations 9, 34 -- Date: Tue, 15 May 2007 08:08:41 -0500 9, 34 -- Message-ID: {8809C39726DC5F43824A3D3BE5D0286A0C868313-at-nashbsasebe01.nr.ad.newellco.com} 9, 34 -- X-MS-Has-Attach: 9, 34 -- X-MS-TNEF-Correlator: 9, 34 -- Thread-Topic: High-magnification imaging help and recommendations 9, 34 -- Thread-Index: AceW8b9UX12A6aMCTM2NqlyFcBlw0QAADgrg 9, 34 -- From: "Zonis, Robert" {Robert.Zonis-at-sanford.com} 9, 34 -- To: {Microscopy-at-microscopy.com} 9, 34 -- X-OriginalArrivalTime: 15 May 2007 13:08:44.0328 (UTC) FILETIME=[2A89E280:01C796F2] 9, 34 -- Content-Transfer-Encoding: 8bit 9, 34 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4FD8kf9003197 ==============================End of - Headers==============================
==============================Original Headers============================== 18, 36 -- From mager-at-interchange.ubc.ca Tue May 15 12:58:40 2007 18, 36 -- Received: from mr7.mail-relay.ubc.ca (gladstone.mail-relay.ubc.ca [137.82.45.27]) 18, 36 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4FHweEX012050 18, 36 -- for {microscopy-at-microscopy.com} ; Tue, 15 May 2007 12:58:40 -0500 18, 36 -- Received: from mr7.mail-relay.ubc.ca (localhost [127.0.0.1]) 18, 36 -- by localhost (Postfix) with SMTP id 9DE1812C36 18, 36 -- for {microscopy-at-microscopy.com} ; Tue, 15 May 2007 10:58:39 -0700 (PDT) 18, 36 -- Received: from mta1.interchange.ubc.ca (mta1.interchange.ubc.ca [142.103.145.69]) 18, 36 -- by mr7.mail-relay.ubc.ca (Postfix) with ESMTP 18, 36 -- for {microscopy-at-microscopy.com} ; Tue, 15 May 2007 10:58:38 -0700 (PDT) 18, 36 -- Received: from Mary (desk.staff.mmat.ubc.ca [137.82.16.178]) 18, 36 -- by smtp.interchange.ubc.ca 18, 36 -- (iPlanet Messaging Server 5.2 HotFix 1.21 (built Sep 8 2003)) 18, 36 -- with ESMTPS id {0JI3000QLGLQ4G-at-smtp.interchange.ubc.ca} for 18, 36 -- microscopy-at-microscopy.com; Tue, 15 May 2007 10:58:38 -0700 (PDT) 18, 36 -- Date: Tue, 15 May 2007 10:58:07 -0700 18, 36 -- From: Mary Mager {mager-at-interchange.ubc.ca} 18, 36 -- Subject: RE: [Microscopy] High-magnification imaging help and recommendations 18, 36 -- In-reply-to: {200705151313.l4FDDSRK012039-at-ns.microscopy.com} 18, 36 -- To: Robert.Zonis-at-sanford.com 18, 36 -- Cc: "'Microscopy'" {microscopy-at-microscopy.com} 18, 36 -- Reply-to: mager-at-interchange.ubc.ca 18, 36 -- Message-id: {0JI3000QMGLQ4G-at-smtp.interchange.ubc.ca} 18, 36 -- Organization: Materials Eng. UBC 18, 36 -- MIME-version: 1.0 18, 36 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 18, 36 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 18, 36 -- Content-type: text/plain; charset=iso-8859-1 18, 36 -- Thread-index: AceW8tQ3Tson15MWR1ilugSqslG8LQAJpoHQ 18, 36 -- X-UBC-Scanned: Sophos PureMessage 5.3.1.294258, Antispam-Engine: 2.5.1.298604, Antispam-Data: 2007.5.15.104435 18, 36 -- X-UBC-Relayed: Relayed through mail-relay.ubc.ca 18, 36 -- X-PerlMx-Spam: Probability=7%, Report=__C230066_P5 0, __CP_MEDIA_BODY 0, __CP_URI_IN_BODY 0, __CT 0, __CTE 0, __CT_TEXT_PLAIN 0, __FRAUD_419_REFNUM 0, __HAS_MSGID 0, __HAS_X_MAILER 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0, __STOCK_PHRASE_24 0, __USER_AGENT_MS_GENERIC 0 18, 36 -- X-Spam-Level: 18, 36 -- X-Spam-Flag: No 18, 36 -- Content-Transfer-Encoding: 8bit 18, 36 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4FHweEX012050 ==============================End of - Headers==============================
What a hoot! Your message made me LOL...I'll be chuckling about that for the rest of the day;-) Thanks!
On May 15, 2007, at 1:59 PM, mager-at-interchange.ubc.ca wrote:
} } } } ---------------------------------------------------------------------- } ------ } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } Dear Robert, } No one else has answered your question, so I will try. I think your } best bet } would be to image the surface of your balls by Scanning Electron } Microscopy } (SEM). This should be available in the closest university or at } most failure } analysis consulting engineering firms. SEMs are fairly common and your } requirements are fairly simple. The SEM should be able to go from } 50 times } magnification to 10,000 times with a good depth of focus, so you } can see the } details on the rounded surface. You should be able to get some good } images } in less than an hour. } Good luck. } } Mary Fletcher (nee Mager) } Electron Microscopist } Materials Eng. UBC } #309 - 6350 Stores Road } Vancouver, B.C. V6T 1Z4 } Canada } Tel: 604-822-5648 } Fax: 604-822-3619 } email: mager-at-interchange.ubc.ca } } -----Original Message----- } X-from: Robert.Zonis-at-sanford.com [mailto:Robert.Zonis-at-sanford.com] } Sent: May 15, 2007 6:13 AM } To: mager-at-interchange.ubc.ca } Subject: [Microscopy] High-magnification imaging help and } recommendations } } } } } ---------------------------------------------------------------------- } ------ } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } } To all: } } I'd like to get some high-magnification images of the 1.00mm tungsten } carbide balls that we use in our ball point and roller ball pens. } The images } I've been able to get with light microscopy do not show enough surface } detail. Can anyone recommend the best type of imaging for this } study and a } facility that will do that sort of imaging for us inexpensively? } } Thanks very much, } } Robert Zonis } Product Development Chemist, LMTC } Sanford L.P. - A Newell Rubbermaid Company } 3 Sharpie Way } Shelbyville, TN 37160 } Direct: +1 (931) 685-6635 } Fax: +1 (931) 685-6613 } robert.zonis-at-sanford.com } www.papermate.com } } } This message may contain information that is confidential and/or } protected } by law. If the reader of this message is not the intended } recipient, you are } hereby notified that any dissemination, distribution, copying or } communication of this message is strictly prohibited. If you have } received } this communication in error, please contact the sender immediately and } delete the message. Please note that although we will take all } commercially } reasonable efforts to prevent viruses from being transmitted from our } systems, it is the responsibility of the recipient to check for and } prevent } adverse action by viruses on its own systems. } } ______________________________________________________________________ } This email has been scanned by the MessageLabs Email Security System. } For more information please visit http://www.messagelabs.com/email } ______________________________________________________________________ } } } ==============================Original } Headers============================== } 9, 34 -- From Robert.Zonis-at-sanford.com Tue May 15 08:08:47 2007 } 9, 34 -- Received: from mail51.messagelabs.com (mail51.messagelabs.com } [216.82.244.51]) } 9, 34 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id } l4FD8kf9003197 } 9, 34 -- for {Microscopy-at-microscopy.com} ; Tue, 15 May 2007 08:08:47 } -0500 } 9, 34 -- X-VirusChecked: Checked } 9, 34 -- X-Env-Sender: Robert.Zonis-at-sanford.com } 9, 34 -- X-Msg-Ref: server-11.tower-51.messagelabs.com!1179234524! } 43756649!1 } 9, 34 -- X-StarScan-Version: 5.5.10.7.1; banners=sanford.com,-,- } 9, 34 -- X-Originating-IP: [12.2.115.11] } 9, 34 -- Received: (qmail 17956 invoked from network); 15 May 2007 } 13:08:44 } -0000 } 9, 34 -- Received: from gnat.newellco.com (HELO } nafepncsvpout2.nr.ad.newellco.com) (12.2.115.11) } 9, 34 -- by server-11.tower-51.messagelabs.com with SMTP; 15 May } 2007 } 13:08:44 -0000 } 9, 34 -- Received: from nafepncsefe02.nr.ad.newellco.com } (unverified) by } nafepncsvpout2.nr.ad.newellco.com } 9, 34 -- (Content Technologies SMTPRS 4.3.17) with ESMTP id } {T7f9b30f9650a059a2116e4-at-nafepncsvpout2.nr.ad.newellco.com} for } {Microscopy-at-microscopy.com} ; } 9, 34 -- Tue, 15 May 2007 08:08:44 -0500 } 9, 34 -- Received: from nashbsasebe01.nr.ad.newellco.com } ([10.217.223.198]) } by nafepncsefe02.nr.ad.newellco.com with Microsoft SMTPSVC } (6.0.3790.1830); } 9, 34 -- Tue, 15 May 2007 08:08:44 -0500 } 9, 34 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 } 9, 34 -- Content-class: urn:content-classes:message } 9, 34 -- MIME-Version: 1.0 } 9, 34 -- Content-Type: text/plain; } 9, 34 -- charset="iso-8859-1" } 9, 34 -- Subject: High-magnification imaging help and recommendations } 9, 34 -- Date: Tue, 15 May 2007 08:08:41 -0500 } 9, 34 -- Message-ID: } {8809C39726DC5F43824A3D3BE5D0286A0C868313-at-nashbsasebe01.nr.ad.newellco } .com} } 9, 34 -- X-MS-Has-Attach: } 9, 34 -- X-MS-TNEF-Correlator: } 9, 34 -- Thread-Topic: High-magnification imaging help and } recommendations } 9, 34 -- Thread-Index: AceW8b9UX12A6aMCTM2NqlyFcBlw0QAADgrg } 9, 34 -- From: "Zonis, Robert" {Robert.Zonis-at-sanford.com} } 9, 34 -- To: {Microscopy-at-microscopy.com} } 9, 34 -- X-OriginalArrivalTime: 15 May 2007 13:08:44.0328 (UTC) } FILETIME=[2A89E280:01C796F2] } 9, 34 -- Content-Transfer-Encoding: 8bit } 9, 34 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } ns.microscopy.com id l4FD8kf9003197 } ==============================End of - } Headers============================== } } } } ==============================Original } Headers============================== } 18, 36 -- From mager-at-interchange.ubc.ca Tue May 15 12:58:40 2007 } 18, 36 -- Received: from mr7.mail-relay.ubc.ca (gladstone.mail- } relay.ubc.ca [137.82.45.27]) } 18, 36 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id l4FHweEX012050 } 18, 36 -- for {microscopy-at-microscopy.com} ; Tue, 15 May 2007 } 12:58:40 -0500 } 18, 36 -- Received: from mr7.mail-relay.ubc.ca (localhost [127.0.0.1]) } 18, 36 -- by localhost (Postfix) with SMTP id 9DE1812C36 } 18, 36 -- for {microscopy-at-microscopy.com} ; Tue, 15 May 2007 } 10:58:39 -0700 (PDT) } 18, 36 -- Received: from mta1.interchange.ubc.ca } (mta1.interchange.ubc.ca [142.103.145.69]) } 18, 36 -- by mr7.mail-relay.ubc.ca (Postfix) with ESMTP } 18, 36 -- for {microscopy-at-microscopy.com} ; Tue, 15 May 2007 } 10:58:38 -0700 (PDT) } 18, 36 -- Received: from Mary (desk.staff.mmat.ubc.ca [137.82.16.178]) } 18, 36 -- by smtp.interchange.ubc.ca } 18, 36 -- (iPlanet Messaging Server 5.2 HotFix 1.21 (built Sep 8 } 2003)) } 18, 36 -- with ESMTPS id {0JI3000QLGLQ4G-at-smtp.interchange.ubc.ca} for } 18, 36 -- microscopy-at-microscopy.com; Tue, 15 May 2007 10:58:38 } -0700 (PDT) } 18, 36 -- Date: Tue, 15 May 2007 10:58:07 -0700 } 18, 36 -- From: Mary Mager {mager-at-interchange.ubc.ca} } 18, 36 -- Subject: RE: [Microscopy] High-magnification imaging help } and recommendations } 18, 36 -- In-reply-to: {200705151313.l4FDDSRK012039-at-ns.microscopy.com} } 18, 36 -- To: Robert.Zonis-at-sanford.com } 18, 36 -- Cc: "'Microscopy'" {microscopy-at-microscopy.com} } 18, 36 -- Reply-to: mager-at-interchange.ubc.ca } 18, 36 -- Message-id: {0JI3000QMGLQ4G-at-smtp.interchange.ubc.ca} } 18, 36 -- Organization: Materials Eng. 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********************************************************************** Beth Richardson Electron Microscopy Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
http://www.plantbio.uga.edu/emlab/
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==============================Original Headers============================== 12, 21 -- From beth-at-plantbio.uga.edu Tue May 15 13:09:41 2007 12, 21 -- Received: from dogwood.plantbio.uga.edu (dogwood.plantbio.uga.edu [128.192.26.2]) 12, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4FI9f2F023772 12, 21 -- for {microscopy-at-microscopy.com} ; Tue, 15 May 2007 13:09:41 -0500 12, 21 -- Received: from [128.192.26.46] ([128.192.26.46]) 12, 21 -- (authenticated user beth-at-plantbio.uga.edu) 12, 21 -- by dogwood.plantbio.uga.edu 12, 21 -- (using TLSv1/SSLv3 with cipher AES128-SHA (128 bits)); 12, 21 -- Tue, 15 May 2007 14:09:38 -0400 12, 21 -- In-Reply-To: {200705151759.l4FHxWXd013259-at-ns.microscopy.com} 12, 21 -- References: {200705151759.l4FHxWXd013259-at-ns.microscopy.com} 12, 21 -- Mime-Version: 1.0 (Apple Message framework v752.3) 12, 21 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 12, 21 -- Message-Id: {24ED5E1C-20A1-403E-9FD0-7D72128815B5-at-plantbio.uga.edu} 12, 21 -- Cc: microscopy microscopy {microscopy-at-microscopy.com} 12, 21 -- Content-Transfer-Encoding: 7bit 12, 21 -- From: Beth Richardson {beth-at-plantbio.uga.edu} 12, 21 -- Subject: Re: [Microscopy] RE: High-magnification imaging help and recommendations 12, 21 -- Date: Tue, 15 May 2007 14:10:17 -0400 12, 21 -- To: mager-at-interchange.ubc.ca 12, 21 -- X-Mailer: Apple Mail (2.752.3) ==============================End of - Headers==============================
You also might consider a stereomicroscope with well-adjusted reflected light. Zeiss' stereomicroscope LUMAR V.12 e.g. has a magnification up to 150x and I know there's a new one coming, the V.20, with magnifications up to 400x (if I remember well). Maybe this could do your job! If you want I could make a few photos for you, but than contact me off-list please! Greets,
Sven Terclavers
-----Original Message----- X-from: mager-at-interchange.ubc.ca [mailto:mager-at-interchange.ubc.ca] Sent: 15 May 2007 20:01 To: sven.terclavers-at-med.kuleuven.be
Dear Robert, No one else has answered your question, so I will try. I think your best bet would be to image the surface of your balls by Scanning Electron Microscopy (SEM). This should be available in the closest university or at most failure analysis consulting engineering firms. SEMs are fairly common and your requirements are fairly simple. The SEM should be able to go from 50 times magnification to 10,000 times with a good depth of focus, so you can see the details on the rounded surface. You should be able to get some good images in less than an hour. Good luck.
-----Original Message----- X-from: Robert.Zonis-at-sanford.com [mailto:Robert.Zonis-at-sanford.com] Sent: May 15, 2007 6:13 AM To: mager-at-interchange.ubc.ca
To all:
I'd like to get some high-magnification images of the 1.00mm tungsten carbide balls that we use in our ball point and roller ball pens. The images I've been able to get with light microscopy do not show enough surface detail. Can anyone recommend the best type of imaging for this study and a facility that will do that sort of imaging for us inexpensively?
Thanks very much,
Robert Zonis Product Development Chemist, LMTC Sanford L.P. - A Newell Rubbermaid Company 3 Sharpie Way Shelbyville, TN 37160 Direct: +1 (931) 685-6635 Fax: +1 (931) 685-6613 robert.zonis-at-sanford.com www.papermate.com
This message may contain information that is confidential and/or protected by law. If the reader of this message is not the intended recipient, you are hereby notified that any dissemination, distribution, copying or communication of this message is strictly prohibited. If you have received this communication in error, please contact the sender immediately and delete the message. Please note that although we will take all commercially reasonable efforts to prevent viruses from being transmitted from our systems, it is the responsibility of the recipient to check for and prevent adverse action by viruses on its own systems.
______________________________________________________________________ This email has been scanned by the MessageLabs Email Security System. For more information please visit http://www.messagelabs.com/email ______________________________________________________________________
==============================Original Headers============================== 9, 34 -- From Robert.Zonis-at-sanford.com Tue May 15 08:08:47 2007 9, 34 -- Received: from mail51.messagelabs.com (mail51.messagelabs.com [216.82.244.51]) 9, 34 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l4FD8kf9003197 9, 34 -- for {Microscopy-at-microscopy.com} ; Tue, 15 May 2007 08:08:47 -0500 9, 34 -- X-VirusChecked: Checked 9, 34 -- X-Env-Sender: Robert.Zonis-at-sanford.com 9, 34 -- X-Msg-Ref: server-11.tower-51.messagelabs.com!1179234524!43756649!1 9, 34 -- X-StarScan-Version: 5.5.10.7.1; banners=sanford.com,-,- 9, 34 -- X-Originating-IP: [12.2.115.11] 9, 34 -- Received: (qmail 17956 invoked from network); 15 May 2007 13:08:44 -0000 9, 34 -- Received: from gnat.newellco.com (HELO nafepncsvpout2.nr.ad.newellco.com) (12.2.115.11) 9, 34 -- by server-11.tower-51.messagelabs.com with SMTP; 15 May 2007 13:08:44 -0000 9, 34 -- Received: from nafepncsefe02.nr.ad.newellco.com (unverified) by nafepncsvpout2.nr.ad.newellco.com 9, 34 -- (Content Technologies SMTPRS 4.3.17) with ESMTP id {T7f9b30f9650a059a2116e4-at-nafepncsvpout2.nr.ad.newellco.com} for {Microscopy-at-microscopy.com} ; 9, 34 -- Tue, 15 May 2007 08:08:44 -0500 9, 34 -- Received: from nashbsasebe01.nr.ad.newellco.com ([10.217.223.198]) by nafepncsefe02.nr.ad.newellco.com with Microsoft SMTPSVC(6.0.3790.1830); 9, 34 -- Tue, 15 May 2007 08:08:44 -0500 9, 34 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 9, 34 -- Content-class: urn:content-classes:message 9, 34 -- MIME-Version: 1.0 9, 34 -- Content-Type: text/plain; 9, 34 -- charset="iso-8859-1" 9, 34 -- Subject: High-magnification imaging help and recommendations 9, 34 -- Date: Tue, 15 May 2007 08:08:41 -0500 9, 34 -- Message-ID: {8809C39726DC5F43824A3D3BE5D0286A0C868313-at-nashbsasebe01.nr.ad.newellco.com} 9, 34 -- X-MS-Has-Attach: 9, 34 -- X-MS-TNEF-Correlator: 9, 34 -- Thread-Topic: High-magnification imaging help and recommendations 9, 34 -- Thread-Index: AceW8b9UX12A6aMCTM2NqlyFcBlw0QAADgrg 9, 34 -- From: "Zonis, Robert" {Robert.Zonis-at-sanford.com} 9, 34 -- To: {Microscopy-at-microscopy.com} 9, 34 -- X-OriginalArrivalTime: 15 May 2007 13:08:44.0328 (UTC) FILETIME=[2A89E280:01C796F2] 9, 34 -- Content-Transfer-Encoding: 8bit 9, 34 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4FD8kf9003197 ==============================End of - Headers==============================
==============================Original Headers============================== 18, 36 -- From mager-at-interchange.ubc.ca Tue May 15 12:58:40 2007 18, 36 -- Received: from mr7.mail-relay.ubc.ca (gladstone.mail-relay.ubc.ca [137.82.45.27]) 18, 36 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4FHweEX012050 18, 36 -- for {microscopy-at-microscopy.com} ; Tue, 15 May 2007 12:58:40 -0500 18, 36 -- Received: from mr7.mail-relay.ubc.ca (localhost [127.0.0.1]) 18, 36 -- by localhost (Postfix) with SMTP id 9DE1812C36 18, 36 -- for {microscopy-at-microscopy.com} ; Tue, 15 May 2007 10:58:39 -0700 (PDT) 18, 36 -- Received: from mta1.interchange.ubc.ca (mta1.interchange.ubc.ca [142.103.145.69]) 18, 36 -- by mr7.mail-relay.ubc.ca (Postfix) with ESMTP 18, 36 -- for {microscopy-at-microscopy.com} ; Tue, 15 May 2007 10:58:38 -0700 (PDT) 18, 36 -- Received: from Mary (desk.staff.mmat.ubc.ca [137.82.16.178]) 18, 36 -- by smtp.interchange.ubc.ca 18, 36 -- (iPlanet Messaging Server 5.2 HotFix 1.21 (built Sep 8 2003)) 18, 36 -- with ESMTPS id {0JI3000QLGLQ4G-at-smtp.interchange.ubc.ca} for 18, 36 -- microscopy-at-microscopy.com; Tue, 15 May 2007 10:58:38 -0700 (PDT) 18, 36 -- Date: Tue, 15 May 2007 10:58:07 -0700 18, 36 -- From: Mary Mager {mager-at-interchange.ubc.ca} 18, 36 -- Subject: RE: [Microscopy] High-magnification imaging help and recommendations 18, 36 -- In-reply-to: {200705151313.l4FDDSRK012039-at-ns.microscopy.com} 18, 36 -- To: Robert.Zonis-at-sanford.com 18, 36 -- Cc: "'Microscopy'" {microscopy-at-microscopy.com} 18, 36 -- Reply-to: mager-at-interchange.ubc.ca 18, 36 -- Message-id: {0JI3000QMGLQ4G-at-smtp.interchange.ubc.ca} 18, 36 -- Organization: Materials Eng. UBC 18, 36 -- MIME-version: 1.0 18, 36 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 18, 36 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 18, 36 -- Content-type: text/plain; charset=iso-8859-1 18, 36 -- Thread-index: AceW8tQ3Tson15MWR1ilugSqslG8LQAJpoHQ 18, 36 -- X-UBC-Scanned: Sophos PureMessage 5.3.1.294258, Antispam-Engine: 2.5.1.298604, Antispam-Data: 2007.5.15.104435 18, 36 -- X-UBC-Relayed: Relayed through mail-relay.ubc.ca 18, 36 -- X-PerlMx-Spam: Probability=7%, Report=__C230066_P5 0, __CP_MEDIA_BODY 0, __CP_URI_IN_BODY 0, __CT 0, __CTE 0, __CT_TEXT_PLAIN 0, __FRAUD_419_REFNUM 0, __HAS_MSGID 0, __HAS_X_MAILER 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0, __STOCK_PHRASE_24 0, __USER_AGENT_MS_GENERIC 0 18, 36 -- X-Spam-Level: 18, 36 -- X-Spam-Flag: No 18, 36 -- Content-Transfer-Encoding: 8bit 18, 36 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4FHweEX012050 ==============================End of - Headers==============================
I agree with Sven with one exception: you might want to try oblique illumination or even darkfield. The off-set illumination will give a shadowing effect that will bring out surface information.
Hope this was helpful, Barbara Foster, President
Microscopy/Microscopy Education 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 W: www.MicroscopyEducation.com
MME is now scheduling customized, on-site courses through July. Call us today for details.
P. S. Need a good general reference or light microscopy text for next semester? Call us today to learn more about "Optimizing LIght Microscopy". Copies still available through MME... even for class-room lots ... and we give quantity discounts. Call Ken Piel at (972)954-8011. At 02:28 PM 5/15/2007, Sven.Terclavers-at-med.kuleuven.be wrote:
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==============================Original Headers============================== 10, 17 -- From bfoster-at-mme1.com Tue May 15 15:45:38 2007 10, 17 -- Received: from mail.plhosting.com (qmail1h.plhosting.com [65.39.254.134]) 10, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4FKjbW3017969 10, 17 -- for {microscopy-at-microscopy.com} ; Tue, 15 May 2007 15:45:37 -0500 10, 17 -- Message-Id: {200705152045.l4FKjbW3017969-at-ns.microscopy.com} 10, 17 -- Received: (qmail 3840 invoked by uid 0); 15 May 2007 20:45:37 -0000 10, 17 -- Received: from unknown (HELO barbsd505.mme1.com) (bfoster-at-mme1.com-at-65.17.57.99) 10, 17 -- by qmail1h.plhosting.com with ESMTPA; 15 May 2007 20:45:36 -0000 10, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 7.1.0.9 10, 17 -- Date: Tue, 15 May 2007 15:45:03 -0500 10, 17 -- To: Sven.Terclavers-at-med.kuleuven.be, microscopy-at-microscopy.com 10, 17 -- From: Barbara Foster {bfoster-at-mme1.com} 10, 17 -- Subject: Re: [Microscopy] High-magnification imaging help and 10, 17 -- In-Reply-To: {200705151858.l4FIwveH007661-at-ns.microscopy.com} 10, 17 -- References: {200705151858.l4FIwveH007661-at-ns.microscopy.com} 10, 17 -- Mime-Version: 1.0 10, 17 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
Dear Robert, I suggest trying Atomic Force Microscopy. This will provide true 3-dimensional information. Furthermore, we have proprietary software that can compute Rk roughness parameters from this data. The Rk parameters are a unique take on roughness and find use when wear and lubrication (liquid supply) are important. Further information about Rk is available at http://www.asmicro.com/software/rk.htm
Disclaimer: we are in business to provide the services and software described above.
regards, Don ============================================= Don Chernoff, Ph.D., President Advanced Surface Microscopy, Inc. E-Mail: donc-at-asmicro.com 3250 N. Post Rd., Ste. 120 Voice: 317-895-5630 INDIANAPOLIS IN 46226 USA Toll free: 800-374-8557 (in USA & Canada) web: http://www.asmicro.com Fax: 317-895-5652 [business activities: analytical services in AFM, AFM probes, consulting, training, calibration and test specimens, calibration and measurement software, used NanoScope equipment.] =============================================
----- Original Message ----- From: Robert.Zonis-at-sanford.com To: donc-at-asmicro.com Sent: Tuesday, May 15, 2007 9:12 AM Subject: [a] [Microscopy] High-magnification imaging help and recommendations
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To all:
I'd like to get some high-magnification images of the 1.00mm tungsten carbide balls that we use in our ball point and roller ball pens. The images I've been able to get with light microscopy do not show enough surface detail. Can anyone recommend the best type of imaging for this study and a facility that will do that sort of imaging for us inexpensively?
Thanks very much,
Robert Zonis Product Development Chemist, LMTC Sanford L.P. - A Newell Rubbermaid Company 3 Sharpie Way Shelbyville, TN 37160 Direct: +1 (931) 685-6635 Fax: +1 (931) 685-6613 robert.zonis-at-sanford.com www.papermate.com
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==============================Original Headers============================== 9, 34 -- From Robert.Zonis-at-sanford.com Tue May 15 08:08:47 2007 9, 34 -- Received: from mail51.messagelabs.com (mail51.messagelabs.com [216.82.244.51]) 9, 34 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l4FD8kf9003197 9, 34 -- for {Microscopy-at-microscopy.com} ; Tue, 15 May 2007 08:08:47 -0500 9, 34 -- X-VirusChecked: Checked 9, 34 -- X-Env-Sender: Robert.Zonis-at-sanford.com 9, 34 -- X-Msg-Ref: server-11.tower-51.messagelabs.com!1179234524!43756649!1 9, 34 -- X-StarScan-Version: 5.5.10.7.1; banners=sanford.com,-,- 9, 34 -- X-Originating-IP: [12.2.115.11] 9, 34 -- Received: (qmail 17956 invoked from network); 15 May 2007 13:08:44 -0000 9, 34 -- Received: from gnat.newellco.com (HELO nafepncsvpout2.nr.ad.newellco.com) (12.2.115.11) 9, 34 -- by server-11.tower-51.messagelabs.com with SMTP; 15 May 2007 13:08:44 -0000 9, 34 -- Received: from nafepncsefe02.nr.ad.newellco.com (unverified) by nafepncsvpout2.nr.ad.newellco.com 9, 34 -- (Content Technologies SMTPRS 4.3.17) with ESMTP id {T7f9b30f9650a059a2116e4-at-nafepncsvpout2.nr.ad.newellco.com} for {Microscopy-at-microscopy.com} ; 9, 34 -- Tue, 15 May 2007 08:08:44 -0500 9, 34 -- Received: from nashbsasebe01.nr.ad.newellco.com ([10.217.223.198]) by nafepncsefe02.nr.ad.newellco.com with Microsoft SMTPSVC(6.0.3790.1830); 9, 34 -- Tue, 15 May 2007 08:08:44 -0500 9, 34 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 9, 34 -- Content-class: urn:content-classes:message 9, 34 -- MIME-Version: 1.0 9, 34 -- Content-Type: text/plain; 9, 34 -- charset="iso-8859-1" 9, 34 -- Subject: High-magnification imaging help and recommendations 9, 34 -- Date: Tue, 15 May 2007 08:08:41 -0500 9, 34 -- Message-ID: {8809C39726DC5F43824A3D3BE5D0286A0C868313-at-nashbsasebe01.nr.ad.newellco.com} 9, 34 -- X-MS-Has-Attach: 9, 34 -- X-MS-TNEF-Correlator: 9, 34 -- Thread-Topic: High-magnification imaging help and recommendations 9, 34 -- Thread-Index: AceW8b9UX12A6aMCTM2NqlyFcBlw0QAADgrg 9, 34 -- From: "Zonis, Robert" {Robert.Zonis-at-sanford.com} 9, 34 -- To: {Microscopy-at-microscopy.com} 9, 34 -- X-OriginalArrivalTime: 15 May 2007 13:08:44.0328 (UTC) FILETIME=[2A89E280:01C796F2] 9, 34 -- Content-Transfer-Encoding: 8bit 9, 34 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4FD8kf9003197 ==============================End of - Headers==============================
==============================Original Headers============================== 23, 22 -- From donc-at-asmicro.com Tue May 15 22:42:56 2007 23, 22 -- Received: from smtp104.sbc.mail.mud.yahoo.com (smtp104.sbc.mail.mud.yahoo.com [68.142.198.203]) 23, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l4G3gtbT010825 23, 22 -- for {microscopy-at-microscopy.com} ; Tue, 15 May 2007 22:42:55 -0500 23, 22 -- Received: (qmail 33567 invoked from network); 16 May 2007 03:42:55 -0000 23, 22 -- Received: from unknown (HELO ASM11) (asmicro-at-sbcglobal.net-at-68.57.205.80 with login) 23, 22 -- by smtp104.sbc.mail.mud.yahoo.com with SMTP; 16 May 2007 03:42:54 -0000 23, 22 -- X-YMail-OSG: _6x_c9UVM1nZzqeaHvOw46YmOjKDlj6azaNht1B.al746Sl5D1O.whr5qMC.tOenBJDmOe0pyIOG_ECoSrKzg5VcKIyE52D3xl2I022iuyTunTdTQ7l.UOyMSdWNibeAnZyHOPLH2yfffilsQlQvFGQdOU6Xy686CMLbL_twAY1K 23, 22 -- Message-ID: {002f01c7976b$ff503590$3301a8c0-at-ASM11} 23, 22 -- From: "Don Chernoff at ASM" {donc-at-asmicro.com} 23, 22 -- To: {Robert.Zonis-at-sanford.com} , "Microscopy List" {microscopy-at-microscopy.com} 23, 22 -- References: {200705151312.l4FDC274008488-at-ns.microscopy.com} 23, 22 -- Subject: Re: [a] [Microscopy] High-magnification imaging help and recommendations 23, 22 -- Date: Tue, 15 May 2007 23:39:41 -0400 23, 22 -- MIME-Version: 1.0 23, 22 -- Content-Type: text/plain; 23, 22 -- charset="iso-8859-1" 23, 22 -- Content-Transfer-Encoding: 7bit 23, 22 -- X-Priority: 3 23, 22 -- X-MSMail-Priority: Normal 23, 22 -- X-Mailer: Microsoft Outlook Express 6.00.2800.1807 23, 22 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1896 ==============================End of - Headers==============================
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Email: mbisher-at-princeton.edu Name: Peggy Bisher
Question: We are doing some spring cleaning and came across some boxes of K-type JEOL filaments. There are 14 of them total.
They are free, to the first person who emails me. Just pay for the postage to mail them.
Margaret(Peggy) E. Bisher Electron Microscopy & Histology Core Facility Manager Department of Molecular Biology Princeton University Moffett Laboratory, Room 113 Princeton, New Jersey Office: (609) 258-7026 Fax: (609) 258-8468 mbisher-at-princeton.edu
Thank you all for your numerous and very instructive answers. I am happy to finally have clear answers and clear solutions to this question.
Best regards, Stephane
____________________________________________________________________________________ 8:00? 8:25? 8:40? Find a flick in no time with the Yahoo! Search movie showtime shortcut. http://tools.search.yahoo.com/shortcuts/#news
==============================Original Headers============================== 4, 20 -- From nizets2-at-yahoo.com Wed May 16 00:01:03 2007 4, 20 -- Received: from web37412.mail.mud.yahoo.com (web37412.mail.mud.yahoo.com [209.191.91.144]) 4, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l4G512Ui020980 4, 20 -- for {microscopy-at-microscopy.com} ; Wed, 16 May 2007 00:01:03 -0500 4, 20 -- Received: (qmail 93530 invoked by uid 60001); 16 May 2007 05:01:02 -0000 4, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 4, 20 -- s=s1024; d=yahoo.com; 4, 20 -- h=X-YMail-OSG:Received:Date:From:Subject:To:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding:Message-ID; 4, 20 -- b=a7Dr2MosG5WEaBvx9gLw5m5warO6IZVF/O98NX+hE/mXVGd1LZ71qwHi9k33Jg8YDBYKT09wlG7gsKBtdFRw8d7gU3bcsS9tzULFSxbS2jsXSmMcjASrYBvYXC4h6zZ1k1Al4gYUoewMoEH4C8Ap9usKw4mKnVHPj09DpGHGhLE=; 4, 20 -- X-YMail-OSG: 2rqpr_sVM1nzI.R91F3HUiCH4Iz3tGl7cp2pS6_VNLLNvxAL3HdjEFQFM63_945Yn1eW0UKBwJcy8dAO_GNXTrbpiQ3u5O_j57.UDsx3NlhL_Q1GRBATu3p1EW0A2w-- 4, 20 -- Received: from [80.121.56.195] by web37412.mail.mud.yahoo.com via HTTP; Tue, 15 May 2007 22:01:02 PDT 4, 20 -- Date: Tue, 15 May 2007 22:01:02 -0700 (PDT) 4, 20 -- From: Stephane Nizet {nizets2-at-yahoo.com} 4, 20 -- Subject: [Microscopy] TEM : objective aperture and EDX - THANKS!! 4, 20 -- To: microscopy-at-microscopy.com 4, 20 -- In-Reply-To: {0JHI008H8Z6UJO-at-smtp.interchange.ubc.ca} 4, 20 -- MIME-Version: 1.0 4, 20 -- Content-Type: text/plain; charset=iso-8859-1 4, 20 -- Content-Transfer-Encoding: 8bit 4, 20 -- Message-ID: {404935.92836.qm-at-web37412.mail.mud.yahoo.com} ==============================End of - Headers==============================
I have tried all types of optical and atomic force microscopes for this sort of thing and they are all plagued a limited depth of field (or just aren't quite as good as SEM). You might even get better images using an SLR digital camera and extension rings rather than a cheap stereo microscope simply because you can easily turn the F stop down to 22.
However, really the only way to go is a scanning electron microscope (SEM). The resolution is in nanometres and the incredible depth of field simply puts it in another league (and I a light microscopist by trade). I remember viewing a burnt pulverised fuel ash cenosphere and marvelling at the surface detail where you could clearly see the crater holes where volatile material had burnt off from inside. Turning the cenosphere round we found that it has split in half and you could see the multi micro-channels left in the carbon residue leading to surface craters. Likewise pollen & hard bodied insects look fantastic. You have the perfect sample, it's dead, hard, unaffected by vacuum and even conducts, you simply have to fix them to an SEM stub and sputter coat. You don't even have any colour in the sample to worry about. And you can get elemental analyses and size data from the specimens (but it costs more and you probably know that already).
I would have thought any local university/college (ideally a materials department) would image your biro balls using SEM for under $200, as it's only a few hours work if you only have a few samples. Ring round if money is tight as prices do vary. The SEM will have dedicated support staff to help you. There are also many small technical support companies who will do this type of SEM work. You probably don't need a sophisticated SEM, which will keep the cost down. I've tried atomic force etc.. and for surface detail, & if your sample can stand it, nothing compares.
Keith
--------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Institute of Ophthalmology UCL, London EC1V 9EL
-----Original Message----- X-from: donc-at-asmicro.com [mailto:donc-at-asmicro.com] Sent: 16 May 2007 04:50 To: keith.morris-at-ucl.ac.uk
Dear Robert, I suggest trying Atomic Force Microscopy. This will provide true 3-dimensional information. Furthermore, we have proprietary software that can compute Rk roughness parameters from this data. The Rk parameters are a unique take on roughness and find use when wear and lubrication (liquid supply) are important. Further information about Rk is available at http://www.asmicro.com/software/rk.htm
Disclaimer: we are in business to provide the services and software described above.
regards, Don ============================================= Don Chernoff, Ph.D., President Advanced Surface Microscopy, Inc. E-Mail: donc-at-asmicro.com 3250 N. Post Rd., Ste. 120 Voice: 317-895-5630 INDIANAPOLIS IN 46226 USA Toll free: 800-374-8557 (in USA & Canada) web: http://www.asmicro.com Fax: 317-895-5652 [business activities: analytical services in AFM, AFM probes, consulting, training, calibration and test specimens, calibration and measurement software, used NanoScope equipment.] =============================================
----- Original Message ----- From: Robert.Zonis-at-sanford.com To: donc-at-asmicro.com Sent: Tuesday, May 15, 2007 9:12 AM Subject: [a] [Microscopy] High-magnification imaging help and recommendations
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To all:
I'd like to get some high-magnification images of the 1.00mm tungsten carbide balls that we use in our ball point and roller ball pens. The images I've been able to get with light microscopy do not show enough surface detail. Can anyone recommend the best type of imaging for this study and a facility that will do that sort of imaging for us inexpensively?
Thanks very much,
Robert Zonis Product Development Chemist, LMTC Sanford L.P. - A Newell Rubbermaid Company 3 Sharpie Way Shelbyville, TN 37160 Direct: +1 (931) 685-6635 Fax: +1 (931) 685-6613 robert.zonis-at-sanford.com www.papermate.com
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==============================Original Headers============================== 9, 34 -- From Robert.Zonis-at-sanford.com Tue May 15 08:08:47 2007 9, 34 -- Received: from mail51.messagelabs.com (mail51.messagelabs.com [216.82.244.51]) 9, 34 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l4FD8kf9003197 9, 34 -- for {Microscopy-at-microscopy.com} ; Tue, 15 May 2007 08:08:47 -0500 9, 34 -- X-VirusChecked: Checked 9, 34 -- X-Env-Sender: Robert.Zonis-at-sanford.com 9, 34 -- X-Msg-Ref: server-11.tower-51.messagelabs.com!1179234524!43756649!1 9, 34 -- X-StarScan-Version: 5.5.10.7.1; banners=sanford.com,-,- 9, 34 -- X-Originating-IP: [12.2.115.11] 9, 34 -- Received: (qmail 17956 invoked from network); 15 May 2007 13:08:44 -0000 9, 34 -- Received: from gnat.newellco.com (HELO nafepncsvpout2.nr.ad.newellco.com) (12.2.115.11) 9, 34 -- by server-11.tower-51.messagelabs.com with SMTP; 15 May 2007 13:08:44 -0000 9, 34 -- Received: from nafepncsefe02.nr.ad.newellco.com (unverified) by nafepncsvpout2.nr.ad.newellco.com 9, 34 -- (Content Technologies SMTPRS 4.3.17) with ESMTP id {T7f9b30f9650a059a2116e4-at-nafepncsvpout2.nr.ad.newellco.com} for {Microscopy-at-microscopy.com} ; 9, 34 -- Tue, 15 May 2007 08:08:44 -0500 9, 34 -- Received: from nashbsasebe01.nr.ad.newellco.com ([10.217.223.198]) by nafepncsefe02.nr.ad.newellco.com with Microsoft SMTPSVC(6.0.3790.1830); 9, 34 -- Tue, 15 May 2007 08:08:44 -0500 9, 34 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 9, 34 -- Content-class: urn:content-classes:message 9, 34 -- MIME-Version: 1.0 9, 34 -- Content-Type: text/plain; 9, 34 -- charset="iso-8859-1" 9, 34 -- Subject: High-magnification imaging help and recommendations 9, 34 -- Date: Tue, 15 May 2007 08:08:41 -0500 9, 34 -- Message-ID: {8809C39726DC5F43824A3D3BE5D0286A0C868313-at-nashbsasebe01.nr.ad.newellco.com} 9, 34 -- X-MS-Has-Attach: 9, 34 -- X-MS-TNEF-Correlator: 9, 34 -- Thread-Topic: High-magnification imaging help and recommendations 9, 34 -- Thread-Index: AceW8b9UX12A6aMCTM2NqlyFcBlw0QAADgrg 9, 34 -- From: "Zonis, Robert" {Robert.Zonis-at-sanford.com} 9, 34 -- To: {Microscopy-at-microscopy.com} 9, 34 -- X-OriginalArrivalTime: 15 May 2007 13:08:44.0328 (UTC) FILETIME=[2A89E280:01C796F2] 9, 34 -- Content-Transfer-Encoding: 8bit 9, 34 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4FD8kf9003197 ==============================End of - Headers==============================
==============================Original Headers============================== 23, 22 -- From donc-at-asmicro.com Tue May 15 22:42:56 2007 23, 22 -- Received: from smtp104.sbc.mail.mud.yahoo.com (smtp104.sbc.mail.mud.yahoo.com [68.142.198.203]) 23, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l4G3gtbT010825 23, 22 -- for {microscopy-at-microscopy.com} ; Tue, 15 May 2007 22:42:55 -0500 23, 22 -- Received: (qmail 33567 invoked from network); 16 May 2007 03:42:55 -0000 23, 22 -- Received: from unknown (HELO ASM11) (asmicro-at-sbcglobal.net-at-68.57.205.80 with login) 23, 22 -- by smtp104.sbc.mail.mud.yahoo.com with SMTP; 16 May 2007 03:42:54 -0000 23, 22 -- X-YMail-OSG: _6x_c9UVM1nZzqeaHvOw46YmOjKDlj6azaNht1B.al746Sl5D1O.whr5qMC.tOenBJDmOe0pyIOG _ECoSrKzg5VcKIyE52D3xl2I022iuyTunTdTQ7l.UOyMSdWNibeAnZyHOPLH2yfffilsQlQvFGQd OU6Xy686CMLbL_twAY1K 23, 22 -- Message-ID: {002f01c7976b$ff503590$3301a8c0-at-ASM11} 23, 22 -- From: "Don Chernoff at ASM" {donc-at-asmicro.com} 23, 22 -- To: {Robert.Zonis-at-sanford.com} , "Microscopy List" {microscopy-at-microscopy.com} 23, 22 -- References: {200705151312.l4FDC274008488-at-ns.microscopy.com} 23, 22 -- Subject: Re: [a] [Microscopy] High-magnification imaging help and recommendations 23, 22 -- Date: Tue, 15 May 2007 23:39:41 -0400 23, 22 -- MIME-Version: 1.0 23, 22 -- Content-Type: text/plain; 23, 22 -- charset="iso-8859-1" 23, 22 -- Content-Transfer-Encoding: 7bit 23, 22 -- X-Priority: 3 23, 22 -- X-MSMail-Priority: Normal 23, 22 -- X-Mailer: Microsoft Outlook Express 6.00.2800.1807 23, 22 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1896 ==============================End of - Headers==============================
==============================Original Headers============================== 38, 23 -- From keith.morris-at-ucl.ac.uk Wed May 16 07:34:28 2007 38, 23 -- Received: from smtp4.global.net.uk (smtp4.global.net.uk [80.189.92.92]) 38, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4GCYRUm031684 38, 23 -- for {Microscopy-at-microscopy.com} ; Wed, 16 May 2007 07:34:28 -0500 38, 23 -- Received: from 80.242.adsl.brightview.com ([80.189.242.80] helo=loungepc) 38, 23 -- by smtp4.global.net.uk with esmtp (Exim 4.42) 38, 23 -- id 1HoIiM-0007kd-QT; Wed, 16 May 2007 13:34:47 +0100 38, 23 -- From: "Keith Morris" {keith.morris-at-ucl.ac.uk} 38, 23 -- To: {donc-at-asmicro.com} 38, 23 -- Cc: {Microscopy-at-microscopy.com} 38, 23 -- References: {200705160349.l4G3nYqW024460-at-ns.microscopy.com} 38, 23 -- Subject: RE: [Microscopy] Re: [a] High-magnification imaging help and recommendations 38, 23 -- Date: Wed, 16 May 2007 13:34:33 +0100 38, 23 -- Message-ID: {001501c797b6$8ed7fc30$0401a8c0-at-loungepc} 38, 23 -- MIME-Version: 1.0 38, 23 -- Content-Type: text/plain; 38, 23 -- charset="us-ascii" 38, 23 -- Content-Transfer-Encoding: 7bit 38, 23 -- X-Mailer: Microsoft Office Outlook 11 38, 23 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 38, 23 -- Thread-Index: AceXbTr5BefqiHNsTWuJqCSca4hNxwAQ0Bkg 38, 23 -- In-Reply-To: {200705160349.l4G3nYqW024460-at-ns.microscopy.com} 38, 23 -- Authenticated-Sender: ==============================End of - Headers==============================
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Email: xiuhuixin-at-yahoo.com.cn Name: Huixin Xiu
Title-Subject: [Filtered] STEM-EDX
Question: Dear all,
I have a weird problem with my STEM-EDX results. The sample is a FIB-prepared multi-layered cross-sectional sample. The structure of the sample is around 800nm Au, 50nm Ni, 50nm Au and 50nm Ni and also 500nm AlGaN. When I put the probe (the diametre about 2nm) in the Au/Ni area, there were very obvious Ga peaks. I thought it was due to X-ray scattering everywhere, but the problem is that the Ga K peak is spuriously strong in Au/Ni area compared with K peak. It seems that the Ga K intensity follows the trend of Au L intensity, but the Ga L intensity does not. When the probe was in AlGaN area, this phenomenon was not present. Could anybody kindly help me to explain why? Many thanks.
The key bit of information is in the first line of your description... "a FIB-prepared..." The FIBs use a Ga ion source. You are seeing the ion implantation which always occurs (to a greater or lesser extent) in every FIB sputtered foil.
I generally see ~1% Ga in most of my ex-situ liftout samples which have been milled at 30kV. Redeposition of sputtered material can contain up to 30% Ga.
Cheers, Henk
At 08:59 AM 05/16/07, you wrote:
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Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://www.ceof.ohio-state.edu Time is that quality of nature which keeps events from happening all at once. Lately it doesn't seem to be working.
==============================Original Headers============================== 12, 26 -- From colijn.1-at-osu.edu Wed May 16 09:26:59 2007 12, 26 -- Received: from ER6S1.ENG.OHIO-STATE.EDU (er6s1.ecr6.ohio-state.edu [164.107.76.2]) 12, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4GEQxf7025201 12, 26 -- for {microscopy-at-microscopy.com} ; Wed, 16 May 2007 09:26:59 -0500 12, 26 -- Received: from CONVERSION-DAEMON.er6s1.eng.ohio-state.edu by 12, 26 -- er6s1.eng.ohio-state.edu (PMDF V6.2-1x11 #31056) 12, 26 -- id {01MGNFA5G42OAADV5L-at-er6s1.eng.ohio-state.edu} for 12, 26 -- microscopy-at-microscopy.com; Wed, 16 May 2007 10:26:58 -0400 (EDT) 12, 26 -- Received: from HOC1.ecr6.ohio-state.edu 12, 26 -- (hoc1.ceof.ohio-state.edu [164.107.76.179]) by er6s1.eng.ohio-state.edu 12, 26 -- (PMDF V6.2-1x11 #31056) 12, 26 -- with ESMTPA id {01MGNFA51BXCAAAJFS-at-er6s1.eng.ohio-state.edu} ; Wed, 12, 26 -- 16 May 2007 10:26:58 -0400 (EDT) 12, 26 -- Date: Wed, 16 May 2007 10:27:34 -0400 12, 26 -- From: "Hendrik O. Colijn" {colijn.1-at-osu.edu} 12, 26 -- Subject: Re: [Microscopy] viaWWW: STEM-EDX 12, 26 -- In-reply-to: {200705161259.l4GCx5Uh014326-at-ns.microscopy.com} 12, 26 -- Sender: colijn-at-er6s1.ecr6.ohio-state.edu 12, 26 -- To: xiuhuixin-at-yahoo.com.cn 12, 26 -- Cc: Microscopy Listserver {microscopy-at-microscopy.com} 12, 26 -- Message-id: {7.0.1.0.2.20070516102338.037cb210-at-osu.edu} 12, 26 -- MIME-version: 1.0 12, 26 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 12, 26 -- Content-type: text/plain; charset=us-ascii; format=flowed 12, 26 -- X-Env-From: auth/colijn.1-at-osu.edu 12, 26 -- References: {200705161259.l4GCx5Uh014326-at-ns.microscopy.com} ==============================End of - Headers==============================
You should consider doing a low-energy, low-angle "cleaning" as the last step in the FIB process. This can be effective in removing much of the Ga implanted from previous FIB steps.
Cheers
Roger A. Ristau, PhD Electron Microscopy Specialist Institute of Materials Science 97 North Eagleville Road University of Connecticut Storrs, CT 06269 vox: 860-486-5379 fax: 860-486-4745
} } Email: xiuhuixin-at-yahoo.com.cn } Name: Huixin Xiu } } Title-Subject: [Filtered] STEM-EDX } } Question: Dear all, } } I have a weird problem with my STEM-EDX results. The sample is a FIB-prepared } multi-layered cross-sectional sample. The structure of the sample is around } 800nm Au, 50nm Ni, 50nm Au and 50nm Ni and also 500nm AlGaN. When I put the } probe (the diametre about 2nm) in the Au/Ni area, there were very obvious Ga } peaks. I thought it was due to X-ray scattering everywhere, but the problem is } that the Ga K peak is spuriously strong in Au/Ni area compared with K peak. It } seems that the Ga K intensity follows the trend of Au L intensity, but the Ga } L intensity does not. When the probe was in AlGaN area, this phenomenon was } not present. Could anybody kindly help me to explain why? Many thanks. } } Yours sincerely, } Helen }
==============================Original Headers============================== 6, 17 -- From raristau-at-ims.uconn.edu Wed May 16 11:07:41 2007 6, 17 -- Received: from mail1.uits.uconn.edu (mail1.uits.uconn.edu [137.99.25.203]) 6, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4GG7fv3006624 6, 17 -- for {microscopy-at-microscopy.com} ; Wed, 16 May 2007 11:07:41 -0500 6, 17 -- Received: from [137.99.20.118] (d20h118.public.uconn.edu [137.99.20.118]) 6, 17 -- by mail1.uits.uconn.edu (8.13.6/8.11.6) with ESMTP id l4GG7dqN024947; 6, 17 -- Wed, 16 May 2007 12:07:40 -0400 6, 17 -- User-Agent: Microsoft-Entourage/10.1.4.030702.0 6, 17 -- Date: Wed, 16 May 2007 12:07:37 -0400 6, 17 -- Subject: Re: [Microscopy] viaWWW: STEM-EDX 6, 17 -- From: Roger Ristau {raristau-at-ims.uconn.edu} 6, 17 -- To: {xiuhuixin-at-yahoo.com.cn} , {microscopy-at-microscopy.com} 6, 17 -- Message-ID: {C270A489.2320%raristau-at-ims.uconn.edu} 6, 17 -- In-Reply-To: {200705161302.l4GD2YWG019513-at-ns.microscopy.com} 6, 17 -- Mime-version: 1.0 6, 17 -- Content-type: text/plain; charset="US-ASCII" 6, 17 -- Content-transfer-encoding: 7bit ==============================End of - Headers==============================
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Email: rkisensee-at-ors-labs.com Name: Bob Isensee
Organization: Oneida Research Services, Inc.
Title-Subject: [Filtered] Jeol 820 stage motors
Question: Does anyone know of where we might find replacement x-y stage motors for a JEOL 820 SEM.(new used or bone yard ---anything!) JEOL won't have anything until August and the motor manufacturer doesn't make this style of motor anymore.
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both sbledsoe-at-iupui.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: sbledsoe-at-iupui.edu Name: Sharon Bledsoe
Title-Subject: [Filtered] Need Help From Imaging Experts
Question: Here is my problem: I can only use Mac iMovie to edit my digital video. I can not do this on a PC. ONLY iMovie. To save a single frame from the video, my choices are either PICT or JPEG format.
My Question is: To save an origianl image, which is the best format, PICT or JPEG?
I'm not interested in hearing about Tiffs, BMPs or any other format as that is not one of my choices.
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.org/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both mbisher-at-princeton.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: mbisher-at-princeton.edu Name: Peggy Bisher
Organization: Princeton University
Title-Subject: [Filtered] Free JEOL Tungsten Filaments Are PROMISED
Question: To all of you who wrote and asked for the filaments, thank you. They have been promised. It was nice to get messages from so many interested people, I'm sorry I did not have more to go around....
JPEG with the lowest compression you can get. PICT is a format on the way out (it was never used much anyway). David
On May 16, 2007, at 6:34 PM, sbledsoe-at-iupui.edu wrote:
} } } } ---------------------------------------------------------------------- } ------ } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } This Question/Comment was submitted to the Microscopy Listserver } using the WWW based Form at http://microscopy.com/ } MicroscopyListserver/MLFormMail.html } ---------------------------------------------------------------------- } ----- } Remember this posting is most likely not from a Subscriber, so when } replying } please copy both sbledsoe-at-iupui.edu as well as the MIcroscopy } Listserver } ---------------------------------------------------------------------- } ----- } } Email: sbledsoe-at-iupui.edu } Name: Sharon Bledsoe } } Title-Subject: [Filtered] Need Help From Imaging Experts } } Question: Here is my problem: I can only use Mac iMovie to edit my } digital video. I can not do this on a PC. ONLY iMovie. To save a } single frame from the video, my choices are either PICT or JPEG } format. } } My Question is: To save an origianl image, which is the best } format, PICT or JPEG? } } I'm not interested in hearing about Tiffs, BMPs or any other format } as that is not one of my choices. } } ---------------------------------------------------------------------- } ----- } } ==============================Original } Headers============================== } 7, 11 -- From zaluzec-at-microscopy.com Wed May 16 20:28:23 2007 } 7, 11 -- Received: from [206.69.208.22] (mac22.zaluzec.com } [206.69.208.22]) } 7, 11 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP } id l4H1SMmq032569 } 7, 11 -- for {microscopy-at-microscopy.com} ; Wed, 16 May 2007 } 20:28:22 -0500 } 7, 11 -- Mime-Version: 1.0 } 7, 11 -- Message-Id: {p06240801c27160281d64-at-[206.69.208.22]} } 7, 11 -- Date: Wed, 16 May 2007 20:28:21 -0500 } 7, 11 -- To: microscopy-at-microscopy.com } 7, 11 -- From: sbledsoe-at-iupui.edu (by way of MicroscopyListserver) } 7, 11 -- Subject: viaWWW: Need Help From Imaging Experts } 7, 11 -- Content-Type: text/plain; charset="us-ascii" } ==============================End of - } Headers==============================
==============================Original Headers============================== 5, 22 -- From Elliott-at-arizona.edu Wed May 16 22:15:37 2007 5, 22 -- Received: from smtpgate.email.arizona.edu (frodo.email.arizona.edu [128.196.132.210]) 5, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4H3FbqS003788 5, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 16 May 2007 22:15:37 -0500 5, 22 -- Received: from frodos_amavis (amavis6.email.arizona.edu [10.0.0.209]) 5, 22 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id 11CEA34054 5, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 16 May 2007 20:15:37 -0700 (MST) 5, 22 -- Received: from [192.168.0.30] (doctorelliott.us [70.57.226.104]) 5, 22 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id D43913406C 5, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 16 May 2007 20:15:33 -0700 (MST) 5, 22 -- Mime-Version: 1.0 (Apple Message framework v752.2) 5, 22 -- In-Reply-To: {200705170134.l4H1Y4Ep015000-at-ns.microscopy.com} 5, 22 -- References: {200705170134.l4H1Y4Ep015000-at-ns.microscopy.com} 5, 22 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 5, 22 -- Message-Id: {88560BC0-DBAC-4ADE-8FBE-6BE92B16E601-at-arizona.edu} 5, 22 -- Content-Transfer-Encoding: 7bit 5, 22 -- From: David Elliott {Elliott-at-arizona.edu} 5, 22 -- Subject: Re: [Microscopy] viaWWW: Need Help From Imaging Experts 5, 22 -- Date: Wed, 16 May 2007 20:15:32 -0700 5, 22 -- To: Microscopy ListServer {Microscopy-at-MSA.Microscopy.Com} 5, 22 -- X-Mailer: Apple Mail (2.752.2) 5, 22 -- X-Virus-Scanned: amavisd-new at email.arizona.edu ==============================End of - Headers==============================
Does anyone have any tips for preparing and imaging bacterial flagella at reasonably high resolutions? I need to examine the flagella and measure potential differences in their width, so I'd like to image them at around 100-200kx. The flagella are about 25nm wide.
So far, I've been making cell-free flagella preps and adsorbing this to carbon-coated copper or nickel grids, and then staining with PTA. They don't look too bad, but I'd like to know if there would be better methods for this kind of thing.
Cheers,
-- Scott J. Coutts ------------------------------------------------------------------------- Bacterial Pathogenesis Research Group Department of Microbiology PO Box 53, Monash University, 3800, Australia Phone +61 3 9905 8592, Fax +61 3 9905 4811 -------------------------------------------------------------------------
==============================Original Headers============================== 6, 26 -- From scott.coutts-at-med.monash.edu.au Thu May 17 00:45:48 2007 6, 26 -- Received: from cartman.its.monash.edu.au (cartman.its.monash.edu.au [130.194.13.166]) 6, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4H5jlj4018159 6, 26 -- for {microscopy-at-microscopy.com} ; Thu, 17 May 2007 00:45:48 -0500 6, 26 -- Received: from curly.its.monash.edu.au ([130.194.13.87]) 6, 26 -- by cartman.its.monash.edu.au 6, 26 -- (Sun Java System Messaging Server 6.2-4.03 (built Sep 22 2005)) 6, 26 -- with ESMTP id {0JI60060O804D030-at-cartman.its.monash.edu.au} for 6, 26 -- microscopy-at-microscopy.com; Thu, 17 May 2007 15:45:45 +1000 (EST) 6, 26 -- Received: from curly.its.monash.edu.au (localhost.localdomain [127.0.0.1]) 6, 26 -- by localhost (Postfix) with ESMTP id D2456AB542 for 6, 26 -- {microscopy-at-microscopy.com} ; Thu, 17 May 2007 15:45:45 +1000 (EST) 6, 26 -- Received: from [130.194.200.122] 6, 26 -- (MicrobSCouttsDT.med.monash.edu.au [130.194.200.122]) 6, 26 -- by curly.its.monash.edu.au (Postfix) with ESMTP id BEAE64FB06 for 6, 26 -- {microscopy-at-microscopy.com} ; Thu, 17 May 2007 15:45:45 +1000 (EST) 6, 26 -- Date: Thu, 17 May 2007 15:46:46 +1000 6, 26 -- From: Scott Coutts {scott.coutts-at-med.monash.edu.au} 6, 26 -- Subject: Bacterial flagella 6, 26 -- To: microscopy-at-microscopy.com 6, 26 -- Message-id: {464BEC46.9060906-at-med.monash.edu.au} 6, 26 -- Organization: Monash University 6, 26 -- MIME-version: 1.0 6, 26 -- Content-type: text/plain; charset=ISO-8859-1; format=flowed 6, 26 -- Content-transfer-encoding: 7bit 6, 26 -- User-Agent: Thunderbird 1.5.0.10 (Windows/20070221) ==============================End of - Headers==============================
} Does anyone have any tips for preparing and imaging bacterial } flagella } at reasonably high resolutions? I need to examine the flagella and } measure potential differences in their width, so I'd like to image } them at around 100-200kx. The flagella are about 25nm wide. } } So far, I've been making cell-free flagella preps and adsorbing this } to carbon-coated copper or nickel grids, and then staining with PTA. } They } don't look too bad, but I'd like to know if there would be better } methods for this kind of thing.
Carbon-coated (copper) grids: yes if you just want to know their width, negative staining appears appropriate and a final magnification of 30.000 to 50.000 x (on film, on CCD-camera) should be sufficient. Stain: PTA might be ok, but you may also try other stains in parallel samples, like Uranylacetate or Uranylformate. You also may add some sugar derivatives (glucose, trehalose) for preventing / minimizing the collapse of the flagella upon air-drying. (how much time do you have? how many samples?how much effort do you want to put into this project?) Ultimately, you may try cryo-TEM of unstained samples, this will give you the samples with highest quality.
best regards, Reinhard Rachel
---------------------- PD Dr. Reinhard Rachel Universitaet Regensburg Centre for EM - NWF III - Lehrstuhl fuer Anatomie Universitaetsstr. 31 D-93053 Regensburg - Germany tel +49 941 943 2837, 1720, 1666(TEM) fax +49 941 943 2868 mail reinhard.rachel-at-biologie.uni-r.de office: VKL 3.1.29
==============================Original Headers============================== 6, 25 -- From reinhard.rachel-at-biologie.uni-regensburg.de Thu May 17 05:58:38 2007 6, 25 -- Received: from rrzmta2.rz.uni-regensburg.de (rrzmta2.rz.uni-regensburg.de [194.94.155.53]) 6, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4HAwbxN003556 6, 25 -- for {Microscopy-at-microscopy.com} ; Thu, 17 May 2007 05:58:37 -0500 6, 25 -- Received: from rrzmta2.rz.uni-regensburg.de (localhost [127.0.0.1]) 6, 25 -- by localhost (Postfix) with SMTP id DEDE64F63 6, 25 -- for {Microscopy-at-microscopy.com} ; Thu, 17 May 2007 12:58:40 +0200 (CEST) 6, 25 -- Received: from localhost (donald1.rz.uni-regensburg.de [132.199.4.91]) 6, 25 -- by rrzmta2.rz.uni-regensburg.de (Postfix) with ESMTP id CF2D14EE1 6, 25 -- for {Microscopy-at-microscopy.com} ; Thu, 17 May 2007 12:58:40 +0200 (CEST) 6, 25 -- Received: from dhcp8003.rz.uni-regensburg.de (dhcp8003.rz.uni-regensburg.de [132.199.87.3]) 6, 25 -- by webmail.uni-regensburg.de (IMP) with HTTP 6, 25 -- for {rar04520-at-rrzlic2.uni-regensburg.de} ; Thu, 17 May 2007 12:58:36 +0200 6, 25 -- Message-ID: {1179399516.464c355c0d4b4-at-webmail.uni-regensburg.de} 6, 25 -- Date: Thu, 17 May 2007 12:58:36 +0200 6, 25 -- From: reinhard rachel {reinhard.rachel-at-biologie.uni-regensburg.de} 6, 25 -- To: Microscopy-at-microscopy.com 6, 25 -- Subject: TEM: Bacterial flagella 6, 25 -- References: {200705170546.l4H5kHqp018621-at-ns.microscopy.com} 6, 25 -- In-Reply-To: {200705170546.l4H5kHqp018621-at-ns.microscopy.com} 6, 25 -- MIME-Version: 1.0 6, 25 -- Content-Type: text/plain; charset=ISO-8859-1 6, 25 -- Content-Transfer-Encoding: 8bit 6, 25 -- User-Agent: Internet Messaging Program (IMP) 3.2.8 6, 25 -- X-Originating-IP: 132.199.87.3 ==============================End of - Headers==============================
Hi, I am looking for surplus, excess, unused JEOL 733 through 8600 vintage electron microprobes. If you have one (733, 840, 6300, 8600) that you need to get rid of or are using as an SEM only and want to get rid of the WDS's I would like to speak with you.
Cheers Mike
-- ******************************************************************** Michael M. Cheatham 312 Heroy Geology Laboratory Phone (315)-443-1261 Syracuse University Fax (315)-443-3363 Syracuse, NY 13244-1070
owner of PLASMACHEM-L: http://listserv.syr.edu/archives/plasmachem-l.html owner of XRF-L: http://listserv.syr.edu/archives/xrf-l.html owner of TIMS-L: http://listserv.syr.edu/archives/tims-l.html owner of SIRIS-L: http://listserv.syr.edu/archives/siris-l.html ********************************************************************
==============================Original Headers============================== 6, 18 -- From mmcheath-at-syr.edu Thu May 17 07:25:27 2007 6, 18 -- Received: from mx5.syr.edu (mx5.syr.edu [128.230.18.67]) 6, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4HCPRYW016962 6, 18 -- for {microscopy-at-microscopy.com} ; Thu, 17 May 2007 07:25:27 -0500 6, 18 -- Received: from [128.230.24.156] (mmcheath.syr.edu [128.230.24.156]) 6, 18 -- (authenticated bits=0) 6, 18 -- by mx5.syr.edu (8.13.7/8.13.7) with ESMTP id l4HCPMc6017072 6, 18 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 6, 18 -- for {microscopy-at-microscopy.com} ; Thu, 17 May 2007 08:25:23 -0400 6, 18 -- Mime-Version: 1.0 6, 18 -- Message-Id: {a06240801c271f9603826-at-[128.230.24.156]} 6, 18 -- Date: Thu, 17 May 2007 08:25:21 -0400 6, 18 -- To: "Microscopy Listserver" {microscopy-at-microscopy.com} 6, 18 -- From: Michael Cheatham {mmcheath-at-syr.edu} 6, 18 -- Subject: Excess JEOL 733 through 8600 Electron microprobes 6, 18 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 6, 18 -- X-Scanner: InterScan AntiVirus for Sendmail 6, 18 -- X-Virus-Scanned: by amavisd-new ==============================End of - Headers==============================
} JPEG with the lowest compression you can get. } PICT is a format on the way out (it was never used much anyway).
Although I don't have any problem with this suggestion --- if there were any benefit for the OP to export single frames as PICT (e.g., avoiding lossy compression), there are a number of softwares capable of converting PICT to any format desired (e.g., Photoshop), which would include batch processing.
cheerios :o) michael shaffer
SEM/MLA Research Coordinator {http://www.mun.ca/creait/maf/} Inco Innovation Centre Memorial University St. John's, Newfoundland
} On May 16, 2007, at 6:34 PM, sbledsoe-at-iupui.edu wrote: } } Question: Here is my problem: I can only use Mac iMovie to edit my } } digital video. I can not do this on a PC. ONLY iMovie. To save a } } single frame from the video, my choices are either PICT or JPEG } } format. } } } } My Question is: To save an origianl image, which is the } best format, } } PICT or JPEG? } } } } I'm not interested in hearing about Tiffs, BMPs or any } other format as that is not one of my choices.
==============================Original Headers============================== 7, 20 -- From michael-at-shaffer.net Thu May 17 07:27:29 2007 7, 20 -- Received: from n034.sc1.he.tucows.com (smtpout1440.sc1.he.tucows.com [64.97.157.140]) 7, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4HCRTOL020556 7, 20 -- for {Microscopy-at-microscopy.com} ; Thu, 17 May 2007 07:27:29 -0500 7, 20 -- Received: from roamingwolf (134.153.130.141) by n034.sc1.he.tucows.com (7.2.069.1) (authenticated as michael-at-shaffer.net) 7, 20 -- id 4630CE3D0038447E for Microscopy-at-microscopy.com; Thu, 17 May 2007 12:27:28 +0000 7, 20 -- From: "michael shaffer" {michael-at-shaffer.net} 7, 20 -- To: "MSA Microscopy list" {Microscopy-at-microscopy.com} 7, 20 -- References: {200705170316.l4H3G40D004455-at-ns.microscopy.com} 7, 20 -- Subject: RE: [Microscopy] Re: viaWWW: Need Help From Imaging Experts 7, 20 -- Date: Thu, 17 May 2007 09:57:26 -0230 7, 20 -- Message-ID: {002901c7987e$bb7cd490$8d829986-at-CREAIT.MUN.CA} 7, 20 -- MIME-Version: 1.0 7, 20 -- Content-Type: text/plain; 7, 20 -- charset="US-ASCII" 7, 20 -- Content-Transfer-Encoding: 7bit 7, 20 -- X-Mailer: Microsoft Office Outlook 11 7, 20 -- In-Reply-To: {200705170316.l4H3G40D004455-at-ns.microscopy.com} 7, 20 -- thread-index: AceYMSqqlyd3bRRPRoWJv5vKs1cx/gATN1hA 7, 20 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 ==============================End of - Headers==============================
Scott- Hit Pubmed and look up the pioneering experts' papers, Namba, DeRosier especially (hope your institution has e-Journal access to J Mol Biol all the way back!). I've not read them to see how they compensate for flattening. But in my field of interest, muscle myosin filaments, the classic way to minimize flattening is Tannic Acid followed by UrAc for neg stain, as in (1986) Stewart, M. and Kensler, R. W. Arrangement of myosin heads in relaxed thick filaments from frog skeletal muscle. J. Mol. Biol. 192:831-851. I think a still earlier 1985 paper about scorpion filaments may have introduced the method.
I think Kensler has used that method ever since, certainly in 2ournal of Structural Biology 137, 154-163 (2002) where it is described in detail. I'll send you that paper off-list.
-mike reedy-
At 12:50 AM -0500 5/17/07, scott.coutts-at-med.monash.edu.au wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
-- -mike reedy-
************************ Michael K. Reedy, M.D. Duke Univ. Med. Center Dept. Cell Biology, Box 3011 (for U.S. Mail) 458 Alex Sands Bldg, Research Dr (courier) Durham, NC 27710
This really looks like a job for AFM. The sample prep would be a lot easier (no staining) and the results would be more accurate.
Hope this is helpful,
Barbara Foster, President
Microscopy/Microscopy Education 313 S Jupiter Rd, Suite 100 Allen, TX 75002 P: 972-954-8011 W: www.MicroscopyEducation.com
MME is now scheduling customized, on-site courses through July. Call us today for details.
P. S. Need a good general reference or light microscopy text for next semester? Call us today to learn more about "Optimizing LIght Microscopy". Copies still available through MME... even for class-room lots ... and we give quantity discounts. Call Ken Piel at (972)954-8011.
At 09:57 AM 5/17/2007, scott.coutts-at-med.monash.edu.au wrote:
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==============================Original Headers============================== 14, 17 -- From bfoster-at-mme1.com Thu May 17 10:11:40 2007 14, 17 -- Received: from mail.plhosting.com (qmail1h.plhosting.com [65.39.254.134]) 14, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4HFBecT023247 14, 17 -- for {microscopy-at-microscopy.com} ; Thu, 17 May 2007 10:11:40 -0500 14, 17 -- Message-Id: {200705171511.l4HFBecT023247-at-ns.microscopy.com} 14, 17 -- Received: (qmail 7094 invoked by uid 0); 17 May 2007 15:11:39 -0000 14, 17 -- Received: from unknown (HELO barbsd505.mme1.com) (bfoster-at-mme1.com-at-65.17.57.99) 14, 17 -- by qmail1h.plhosting.com with ESMTPA; 17 May 2007 15:11:38 -0000 14, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 7.1.0.9 14, 17 -- Date: Thu, 17 May 2007 10:11:01 -0500 14, 17 -- To: scott.coutts-at-med.monash.edu.au, microscopy-at-microscopy.com 14, 17 -- From: Barbara Foster {bfoster-at-mme1.com} 14, 17 -- Subject: Re: [Microscopy] Bacterial flagella 14, 17 -- In-Reply-To: {200705170548.l4H5mQj0021929-at-ns.microscopy.com} 14, 17 -- References: {200705170548.l4H5mQj0021929-at-ns.microscopy.com} 14, 17 -- Mime-Version: 1.0 14, 17 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
This may be a bit more fundamental than what you are looking for, but here goes:
If you go to the MSA website, you can find a statement about ethical image processing that in essence says that images need to be stored uncompressed. JPEG is a format that includes lossy compression, and I believe that PICT can also contain lossy compression in the form of JPEG. Both therefore don't seem to be the best choices.
I also found this on the web:
In the next version of its operating system, MacOS X, Apple has decided to replace PICT by PDF. This means that the use of PICT for exchanging data will slowly diminish.
It looks like sooner or later you will have to find a different choice anyway. I would look at the whole chain of acquisition to see if you cannot switch to something else.
Why can't you use anything else? Does it have to do with the original file format that can only be read by iMovie? Or are you tied to the Mac platform?
mike
Michael Bode, Ph.D.
General Manager
OLYMPUS SOFT IMAGING SOLUTIONS 12596 West Bayaud Ave #300 Lakewood, CO 80228 USA Tel.: +1 (303) 234-9270 Fax.: +1 (303) 234-9271 E-mail: Mike.Bode-at-olympus-sis.com www.olympus-sis.com
-----Original Message----- X-from: sbledsoe-at-iupui.edu [mailto:sbledsoe-at-iupui.edu] Sent: Wednesday, May 16, 2007 19:39 To: Mike Bode
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MicroscopyListserver/MLFormMail.html ------------------------------------------------------------------------ --- Remember this posting is most likely not from a Subscriber, so when replying please copy both sbledsoe-at-iupui.edu as well as the MIcroscopy Listserver ------------------------------------------------------------------------ ---
Email: sbledsoe-at-iupui.edu Name: Sharon Bledsoe
Title-Subject: [Filtered] Need Help From Imaging Experts
Question: Here is my problem: I can only use Mac iMovie to edit my digital video. I can not do this on a PC. ONLY iMovie. To save a single frame from the video, my choices are either PICT or JPEG format.
My Question is: To save an origianl image, which is the best format, PICT or JPEG?
I'm not interested in hearing about Tiffs, BMPs or any other format as that is not one of my choices.
Looking for second condenser (C2) low-background 15, 30, 50 and 100 micron apertures for FEI CM300 TEM.
These are not the standard Pt thin foil discs. I got a quote for a set for approx. $8000 from FEI. Any alternative suppliers out there?
Thanks,
Krassimir. _______________________________________ Krassimir N. Bozhilov Central Facility for Advanced Microscopy and Microanalysis University of California Riverside, CA 92521
tel 951 827 2998 fax 951 827 2489 bozhilov-at-ucr.edu _______________________________________
==============================Original Headers============================== 7, 19 -- From bozhilov-at-ucr.edu Thu May 17 11:35:52 2007 7, 19 -- Received: from sentinel.ucr.edu (sentinel.ucr.edu [138.23.226.228]) 7, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4HGZqYO015701 7, 19 -- for {microscopy-at-microscopy.com} ; Thu, 17 May 2007 11:35:52 -0500 7, 19 -- Received: from [138.23.185.162] (igppb116g.ucr.edu [138.23.185.162]) 7, 19 -- by sentinel.ucr.edu (MOS 3.6.6-GR) 7, 19 -- with ESMTP id ELY89098 (AUTH via LOGINBEFORESMTP) 7, 19 -- for {microscopy-at-microscopy.com} ; 7, 19 -- Thu, 17 May 2007 09:35:47 -0700 (PDT) 7, 19 -- Mime-Version: 1.0 (Apple Message framework v752.2) 7, 19 -- Content-Transfer-Encoding: 7bit 7, 19 -- Message-Id: {8A1BE97E-989A-4B5C-A8FB-162874C89F6C-at-ucr.edu} 7, 19 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 7, 19 -- To: microscopy-at-microscopy.com 7, 19 -- From: KN Bozhilov {bozhilov-at-ucr.edu} 7, 19 -- Subject: C2 apertures 7, 19 -- Date: Thu, 17 May 2007 09:35:46 -0700 7, 19 -- X-Mailer: Apple Mail (2.752.2) 7, 19 -- X-Junkmail-Whitelist: YES (by domain whitelist at sentinel.ucr.edu) ==============================End of - Headers==============================
Krassimir; That sounds like typical prices for FIB cut apertures for field emission TEMs, but is enough to make anyone say 'ouch!' I bought some cheap ones from Pella (~$60 as I recall) a little smaller than what I needed, and FIB cut them myself. If you have a FIB, it is pretty easy.
John Mardinly Intel
This is not an opinion of Intel Corporation.
-----Original Message----- X-from: bozhilov-at-ucr.edu [mailto:bozhilov-at-ucr.edu] Sent: Thursday, May 17, 2007 9:36 AM To: Mardinly, John
Looking for second condenser (C2) low-background 15, 30, 50 and 100 micron apertures for FEI CM300 TEM.
These are not the standard Pt thin foil discs. I got a quote for a set for approx. $8000 from FEI. Any alternative suppliers out there?
Thanks,
Krassimir. _______________________________________ Krassimir N. Bozhilov Central Facility for Advanced Microscopy and Microanalysis University of California Riverside, CA 92521
tel 951 827 2998 fax 951 827 2489 bozhilov-at-ucr.edu _______________________________________
==============================Original Headers============================== 7, 19 -- From bozhilov-at-ucr.edu Thu May 17 11:35:52 2007 7, 19 -- Received: from sentinel.ucr.edu (sentinel.ucr.edu [138.23.226.228]) 7, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4HGZqYO015701 7, 19 -- for {microscopy-at-microscopy.com} ; Thu, 17 May 2007 11:35:52 -0500 7, 19 -- Received: from [138.23.185.162] (igppb116g.ucr.edu [138.23.185.162]) 7, 19 -- by sentinel.ucr.edu (MOS 3.6.6-GR) 7, 19 -- with ESMTP id ELY89098 (AUTH via LOGINBEFORESMTP) 7, 19 -- for {microscopy-at-microscopy.com} ; 7, 19 -- Thu, 17 May 2007 09:35:47 -0700 (PDT) 7, 19 -- Mime-Version: 1.0 (Apple Message framework v752.2) 7, 19 -- Content-Transfer-Encoding: 7bit 7, 19 -- Message-Id: {8A1BE97E-989A-4B5C-A8FB-162874C89F6C-at-ucr.edu} 7, 19 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 7, 19 -- To: microscopy-at-microscopy.com 7, 19 -- From: KN Bozhilov {bozhilov-at-ucr.edu} 7, 19 -- Subject: C2 apertures 7, 19 -- Date: Thu, 17 May 2007 09:35:46 -0700 7, 19 -- X-Mailer: Apple Mail (2.752.2) 7, 19 -- X-Junkmail-Whitelist: YES (by domain whitelist at sentinel.ucr.edu) ==============================End of - Headers==============================
==============================Original Headers============================== 16, 38 -- From john.mardinly-at-intel.com Thu May 17 11:49:21 2007 16, 38 -- Received: from mga09.intel.com (mga09.intel.com [134.134.136.24]) 16, 38 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4HGnKlY027440 16, 38 -- for {Microscopy-at-msa.microscopy.com} ; Thu, 17 May 2007 11:49:21 -0500 16, 38 -- Received: from fmsmga002.fm.intel.com ([10.253.24.26]) 16, 38 -- by orsmga102.jf.intel.com with ESMTP; 17 May 2007 09:49:09 -0700 16, 38 -- X-ExtLoop1: 1 16, 38 -- X-IronPort-AV: E=Sophos;i="4.14,549,1170662400"; 16, 38 -- d="scan'208";a="87838105" 16, 38 -- Received: from fmsmsxpoc001.fm.intel.com (HELO fmsmsxpoc001.amr.corp.intel.com) ([132.233.49.22]) 16, 38 -- by fmsmga002.fm.intel.com with ESMTP; 17 May 2007 09:49:10 -0700 16, 38 -- Received: from fmsmsx334.amr.corp.intel.com (132.233.42.1) by 16, 38 -- fmsmsxpoc001.amr.corp.intel.com (132.233.49.22) with Microsoft SMTP Server id 16, 38 -- 8.0.685.24; Thu, 17 May 2007 09:49:09 -0700 16, 38 -- Received: from scsmsx412.amr.corp.intel.com ([10.3.90.31]) by 16, 38 -- fmsmsx334.amr.corp.intel.com with Microsoft SMTPSVC(6.0.3790.1830); Thu, 17 16, 38 -- May 2007 09:49:09 -0700 16, 38 -- Received: from scsmsx415.amr.corp.intel.com ([10.3.90.34]) by 16, 38 -- scsmsx412.amr.corp.intel.com with Microsoft SMTPSVC(6.0.3790.1830); Thu, 17 16, 38 -- May 2007 09:49:09 -0700 16, 38 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 16, 38 -- Content-Class: urn:content-classes:message 16, 38 -- MIME-Version: 1.0 16, 38 -- Content-Type: text/plain; charset="us-ascii" 16, 38 -- Subject: RE: [Microscopy] C2 apertures 16, 38 -- Date: Thu, 17 May 2007 09:49:08 -0700 16, 38 -- Message-ID: {F3CB8931ABF8294DB889E977150CAD687BC457-at-scsmsx415.amr.corp.intel.com} 16, 38 -- In-Reply-To: {200705171635.l4HGZwrY015822-at-ns.microscopy.com} 16, 38 -- X-MS-Has-Attach: 16, 38 -- X-MS-TNEF-Correlator: 16, 38 -- Thread-Topic: [Microscopy] C2 apertures 16, 38 -- Thread-Index: AceYoXQTXT3fB0dnRWSkO11BnFDRzAAAR0SQ 16, 38 -- From: "Mardinly, John" {john.mardinly-at-intel.com} 16, 38 -- To: {bozhilov-at-ucr.edu} 16, 38 -- CC: {Microscopy-at-msa.microscopy.com} 16, 38 -- X-OriginalArrivalTime: 17 May 2007 16:49:09.0188 (UTC) FILETIME=[49FEE040:01C798A3] 16, 38 -- Content-Transfer-Encoding: 8bit 16, 38 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4HGnKlY027440 ==============================End of - Headers==============================
These are not disk foils. They look like a small cylinder with 3 mm diameter and about 5-6 mm height.
_______________________________________ Krassimir N. Bozhilov Central Facility for Advanced Microscopy and Microanalysis University of California Riverside, CA 92521
tel 951 827 2998 fax 951 827 2489 bozhilov-at-ucr.edu _______________________________________
On May 17, 2007, at 9:51 AM, john.mardinly-at-intel.com wrote:
} } } } ---------------------------------------------------------------------- } ------ } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } Krassimir; } That sounds like typical prices for FIB cut apertures for field } emission TEMs, but is enough to make anyone say 'ouch!' I bought some } cheap ones from Pella (~$60 as I recall) a little smaller than what I } needed, and FIB cut them myself. If you have a FIB, it is pretty easy. } } John Mardinly } Intel } } This is not an opinion of Intel Corporation. } } -----Original Message----- } X-from: bozhilov-at-ucr.edu [mailto:bozhilov-at-ucr.edu] } Sent: Thursday, May 17, 2007 9:36 AM } To: Mardinly, John } Subject: [Microscopy] C2 apertures } } } } } ---------------------------------------------------------------------- } -- } ---- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -- } ---- } } Looking for second condenser (C2) low-background 15, 30, 50 and 100 } micron apertures for FEI CM300 TEM. } } These are not the standard Pt thin foil discs. } I got a quote for a set for approx. $8000 from FEI. Any alternative } suppliers out there? } } Thanks, } } Krassimir. } _______________________________________ } Krassimir N. Bozhilov } Central Facility for Advanced Microscopy and Microanalysis } University of California } Riverside, CA 92521 } } tel 951 827 2998 } fax 951 827 2489 } bozhilov-at-ucr.edu } _______________________________________ } } } } ==============================Original } Headers============================== } 7, 19 -- From bozhilov-at-ucr.edu Thu May 17 11:35:52 2007 } 7, 19 -- Received: from sentinel.ucr.edu (sentinel.ucr.edu } [138.23.226.228]) } 7, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id l4HGZqYO015701 } 7, 19 -- for {microscopy-at-microscopy.com} ; Thu, 17 May 2007 } 11:35:52 -0500 } 7, 19 -- Received: from [138.23.185.162] (igppb116g.ucr.edu } [138.23.185.162]) } 7, 19 -- by sentinel.ucr.edu (MOS 3.6.6-GR) } 7, 19 -- with ESMTP id ELY89098 (AUTH via LOGINBEFORESMTP) } 7, 19 -- for {microscopy-at-microscopy.com} ; } 7, 19 -- Thu, 17 May 2007 09:35:47 -0700 (PDT) } 7, 19 -- Mime-Version: 1.0 (Apple Message framework v752.2) } 7, 19 -- Content-Transfer-Encoding: 7bit } 7, 19 -- Message-Id: {8A1BE97E-989A-4B5C-A8FB-162874C89F6C-at-ucr.edu} } 7, 19 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; } format=flowed } 7, 19 -- To: microscopy-at-microscopy.com } 7, 19 -- From: KN Bozhilov {bozhilov-at-ucr.edu} } 7, 19 -- Subject: C2 apertures } 7, 19 -- Date: Thu, 17 May 2007 09:35:46 -0700 } 7, 19 -- X-Mailer: Apple Mail (2.752.2) } 7, 19 -- X-Junkmail-Whitelist: YES (by domain whitelist at } sentinel.ucr.edu) } ==============================End of - } Headers============================== } } } ==============================Original } Headers============================== } 16, 38 -- From john.mardinly-at-intel.com Thu May 17 11:49:21 2007 } 16, 38 -- Received: from mga09.intel.com (mga09.intel.com } [134.134.136.24]) } 16, 38 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id l4HGnKlY027440 } 16, 38 -- for {Microscopy-at-msa.microscopy.com} ; Thu, 17 May 2007 } 11:49:21 -0500 } 16, 38 -- Received: from fmsmga002.fm.intel.com ([10.253.24.26]) } 16, 38 -- by orsmga102.jf.intel.com with ESMTP; 17 May 2007 } 09:49:09 -0700 } 16, 38 -- X-ExtLoop1: 1 } 16, 38 -- X-IronPort-AV: E=Sophos;i="4.14,549,1170662400"; } 16, 38 -- d="scan'208";a="87838105" } 16, 38 -- Received: from fmsmsxpoc001.fm.intel.com (HELO } fmsmsxpoc001.amr.corp.intel.com) ([132.233.49.22]) } 16, 38 -- by fmsmga002.fm.intel.com with ESMTP; 17 May 2007 } 09:49:10 -0700 } 16, 38 -- Received: from fmsmsx334.amr.corp.intel.com } (132.233.42.1) by } 16, 38 -- fmsmsxpoc001.amr.corp.intel.com (132.233.49.22) with } Microsoft SMTP Server id } 16, 38 -- 8.0.685.24; Thu, 17 May 2007 09:49:09 -0700 } 16, 38 -- Received: from scsmsx412.amr.corp.intel.com } ([10.3.90.31]) by } 16, 38 -- fmsmsx334.amr.corp.intel.com with Microsoft SMTPSVC } (6.0.3790.1830); Thu, 17 } 16, 38 -- May 2007 09:49:09 -0700 } 16, 38 -- Received: from scsmsx415.amr.corp.intel.com } ([10.3.90.34]) by } 16, 38 -- scsmsx412.amr.corp.intel.com with Microsoft SMTPSVC } (6.0.3790.1830); Thu, 17 } 16, 38 -- May 2007 09:49:09 -0700 } 16, 38 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 } 16, 38 -- Content-Class: urn:content-classes:message } 16, 38 -- MIME-Version: 1.0 } 16, 38 -- Content-Type: text/plain; charset="us-ascii" } 16, 38 -- Subject: RE: [Microscopy] C2 apertures } 16, 38 -- Date: Thu, 17 May 2007 09:49:08 -0700 } 16, 38 -- Message-ID: } {F3CB8931ABF8294DB889E977150CAD687BC457-at-scsmsx415.amr.corp.intel.com} } 16, 38 -- In-Reply-To: {200705171635.l4HGZwrY015822-at-ns.microscopy.com} } 16, 38 -- X-MS-Has-Attach: } 16, 38 -- X-MS-TNEF-Correlator: } 16, 38 -- Thread-Topic: [Microscopy] C2 apertures } 16, 38 -- Thread-Index: AceYoXQTXT3fB0dnRWSkO11BnFDRzAAAR0SQ } 16, 38 -- From: "Mardinly, John" {john.mardinly-at-intel.com} } 16, 38 -- To: {bozhilov-at-ucr.edu} } 16, 38 -- CC: {Microscopy-at-msa.microscopy.com} } 16, 38 -- X-OriginalArrivalTime: 17 May 2007 16:49:09.0188 (UTC) } FILETIME=[49FEE040:01C798A3] } 16, 38 -- Content-Transfer-Encoding: 8bit } 16, 38 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } ns.microscopy.com id l4HGnKlY027440 } ==============================End of - } Headers==============================
==============================Original Headers============================== 7, 21 -- From bozhilov-at-ucr.edu Thu May 17 12:14:50 2007 7, 21 -- Received: from sentinel.ucr.edu (sentinel.ucr.edu [138.23.226.228]) 7, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4HHEkQr007386 7, 21 -- for {microscopy-at-microscopy.com} ; Thu, 17 May 2007 12:14:49 -0500 7, 21 -- Received: from [138.23.185.162] (igppb116g.ucr.edu [138.23.185.162]) 7, 21 -- by sentinel.ucr.edu (MOS 3.6.6-GR) 7, 21 -- with ESMTP id ELY95914 (AUTH via LOGINBEFORESMTP); 7, 21 -- Thu, 17 May 2007 10:08:03 -0700 (PDT) 7, 21 -- In-Reply-To: {200705171651.l4HGpfus030874-at-ns.microscopy.com} 7, 21 -- References: {200705171651.l4HGpfus030874-at-ns.microscopy.com} 7, 21 -- Mime-Version: 1.0 (Apple Message framework v752.2) 7, 21 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 7, 21 -- Message-Id: {6A414FCC-B6A4-4273-AE52-2D5AE229B5D2-at-ucr.edu} 7, 21 -- Cc: john.mardinly-at-intel.com 7, 21 -- Content-Transfer-Encoding: 7bit 7, 21 -- From: KN Bozhilov {bozhilov-at-ucr.edu} 7, 21 -- Subject: Re: [Microscopy] RE: C2 apertures 7, 21 -- Date: Thu, 17 May 2007 10:08:02 -0700 7, 21 -- To: microscopy-at-microscopy.com 7, 21 -- X-Mailer: Apple Mail (2.752.2) 7, 21 -- X-Junkmail-Whitelist: YES (by domain whitelist at sentinel.ucr.edu) ==============================End of - Headers==============================
Since Ladd produces the vast majority of apertures in the U.S, we surely can make them up for you. Please contact us and we'll discuss materials, hole sizes, etc. To see samples of what we can do see our website at: http://www.laddresearch.com/Key_Products/Apertures/apertures.html
Disclaimer: Ladd has produced EM supplies and a wide variety of apertures for over 50 years.
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==============================Original Headers============================== 12, 27 -- From jd-at-laddresearch.com Thu May 17 12:15:36 2007 12, 27 -- Received: from arrows.electric.net (arrows.electric.net [216.129.90.200]) 12, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4HHFZOh008172 12, 27 -- for {microscopy-at-microscopy.com} ; Thu, 17 May 2007 12:15:35 -0500 12, 27 -- Received: from root by arrows.electric.net with emc1-ok (Exim 4.62) 12, 27 -- (envelope-from {jd-at-laddresearch.com} ) 12, 27 -- id 1HojZe-0000zY-TH; Thu, 17 May 2007 10:15:34 -0700 12, 27 -- Received: by emcmailer; Thu, 17 May 2007 10:15:34 -0700 12, 27 -- Received: from [216.204.198.170] (helo=NewServer.laddresearch.com) 12, 27 -- by arrows.electric.net with esmtps (TLSv1:AES256-SHA:256) 12, 27 -- (Exim 4.62) 12, 27 -- (envelope-from {jd-at-laddresearch.com} ) 12, 27 -- id 1HojZc-0000x9-W9; Thu, 17 May 2007 10:15:33 -0700 12, 27 -- X-Mailer: QUALCOMM Windows Eudora Version 7.1.0.9 12, 27 -- Date: Thu, 17 May 2007 13:14:59 -0400 12, 27 -- To: bozhilov-at-ucr.edu 12, 27 -- From: jd {jd-at-laddresearch.com} 12, 27 -- Subject: Re: [Microscopy] C2 apertures 12, 27 -- Cc: Microscopy listserver {microscopy-at-microscopy.com} 12, 27 -- In-Reply-To: {200705171642.l4HGgJD0026309-at-ns.microscopy.com} 12, 27 -- References: {200705171642.l4HGgJD0026309-at-ns.microscopy.com} 12, 27 -- Mime-Version: 1.0 12, 27 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 12, 27 -- X-Outbound-IP: 216.204.198.170 12, 27 -- X-Env-From: jd-at-laddresearch.com 12, 27 -- X-Virus-Status: Scanned by VirusSMART (c) 12, 27 -- Message-Id: {E1HojZe-0000zY-TH-at-arrows.electric.net} ==============================End of - Headers==============================
The Zatkoff Company (www.zatkoff.com) also carries an extensive stock of O-rings. Before trying to make an O-ring yourself I would recommend checking with them to see if they couldn't furnish something to meet your needs. -- Wilbur C. Bigelow, Professor Emeritus Materials Sci. & Engr., Univ. of Michigan Ann Arbor, Michigan 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-975-0858 Address mail to: 2911 Whittier Court Ann Arbor, MI 48104-6731
==============================Original Headers============================== 1, 14 -- From bigelow-at-umich.edu Thu May 17 14:37:50 2007 1, 14 -- Received: from skycaptain.mr.itd.umich.edu (smtp.mail.umich.edu [141.211.93.160]) 1, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4HJbojt001489 1, 14 -- for {microscopy-at-microscopy.com} ; Thu, 17 May 2007 14:37:50 -0500 1, 14 -- Received: FROM [141.212.131.221] (bigelow-g4.engin.umich.edu [141.212.131.221]) 1, 14 -- BY skycaptain.mr.itd.umich.edu ID 464CAF0D.60D1B.9874 ; 1, 14 -- 17 May 2007 15:37:49 -0400 1, 14 -- Mime-Version: 1.0 1, 14 -- Message-Id: {p06240801c2725ef43d43-at-[141.212.131.221]} 1, 14 -- Date: Thu, 17 May 2007 15:37:47 -0400 1, 14 -- To: Microscopy Listserver {microscopy-at-microscopy.com} 1, 14 -- From: Wil Bigelow {bigelow-at-umich.edu} 1, 14 -- Subject: [Microscopy}RE: O-ring 1, 14 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
One of our students wants to work on 2010F at 120 KV. At this moment the beam is very weak if he follows a technical note from JEOL on our 2010F TEM. He was told he can get bright beam if he has a device called Short switch for 120KV. Local service engineer can install the short switch easily. But JEOL USA told him that the device costs about $11,000.00 and he has to wait for four months for delivery. But he needs it urgently.
Is there any one who has the device and our student may borrow or rent? Or is there any idea to get bright beam at 120KV on 2010F?
Thanks a lot.
Zhenquan Liu
Zhenquan Liu Senior Research Specialist School of Materials Arizona State University 901 S Palm Walk PSA 213 Tempe, 85287-1704 Tel (480) 965 4512 Fax (480) 965 9004
==============================Original Headers============================== 9, 24 -- From Zhenquan.Liu-at-asu.edu Thu May 17 15:58:55 2007 9, 24 -- Received: from epo-int1.asu.edu (epo-int1.asu.edu [129.219.187.20]) 9, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4HKwtC3014720 9, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 17 May 2007 15:58:55 -0500 9, 24 -- Received: from EX03.asurite.ad.asu.edu (excl1-b0.asurite.ad.asu.edu [129.219.12.197]) 9, 24 -- by epo-int1.asu.edu (Switch-3.1.8/Switch-3.1.7/asu-postoffice-prod) with ESMTP id l4HKwprS028596 9, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 17 May 2007 13:58:52 -0700 9, 24 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 9, 24 -- Content-class: urn:content-classes:message 9, 24 -- MIME-Version: 1.0 9, 24 -- Content-Type: text/plain; 9, 24 -- charset="US-ASCII" 9, 24 -- Subject: Short Switch for 120KV on 2010F TEM 9, 24 -- Date: Thu, 17 May 2007 13:58:52 -0700 9, 24 -- Message-ID: {60D5D23B9F20DF4C8DC3991979F9094AFA56A0-at-EX03.asurite.ad.asu.edu} 9, 24 -- X-MS-Has-Attach: 9, 24 -- X-MS-TNEF-Correlator: 9, 24 -- Thread-Topic: Short Switch for 120KV on 2010F TEM 9, 24 -- Thread-Index: AceYxk6zfzgfAr37RTmOMPYxIcZS0w== 9, 24 -- From: "Zhenquan Liu" {Zhenquan.Liu-at-asu.edu} 9, 24 -- To: {Microscopy-at-microscopy.com} 9, 24 -- X-Virus-Scanned: by amavisd-new 9, 24 -- Content-Transfer-Encoding: 8bit 9, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4HKwtC3014720 ==============================End of - Headers==============================
Krassimir; 5-6 mm height? I would suspect that the actual aperture must be inserted in some kind of holder, because it would seem to be difficult to form an aperture from solid material that thick. The Pella web site shows cross-sections of Philips apertures that are 3 mm diameter, and 0.25 mm high, with a taper so that the thickness where the beam goes is much less. However, I don't see anything 5-6 mm high in any catalogs, so if the apertures are one piece, you might be stuck with the FEI deal.
John Mardinly Intel
This is not an opinion of Intel Corp.
-----Original Message----- X-from: KN Bozhilov [mailto:bozhilov-at-ucr.edu] Sent: Thursday, May 17, 2007 10:08 AM To: microscopy-at-microscopy.com Cc: Mardinly, John
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.com/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both abansal-at-ilypsa.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: abansal-at-ilypsa.com Name: Anu
Organization: Ilypsa
Title-Subject: [Filtered] Sectioning polymer beads for optical microscopy
Question: Does anyone have experience sectioning 100 to 200 micron sized polystryrene beads for optical microscopy? What are the best embedding media and microtomy conditions to get good sections?
I am having some sectioning difficulties with cross sections of cultured cells grown on Thermanox coverslips.
The monolayer of cells grown on the coverslips have been processed by the progressive lowering of temperature technique (in an AFS), using ethanol as the dehydration solvent, and then followed by embedding in Lowicryl HM20.
The problem I am having is that the Thermanox coverslips are separating from the Lowicryl HM20 resin block making cross-sectional ultrathin sectioning (and semi-thin for that matter) that includes the coverslip difficult.
The separation is easily seen through the binocular head of the ultramicrotome even before I start trimming the block down. The resin simply hasn't 'bonded' to the coverslip. (I am able to remove the coverslip from the block and then get good ultrathin sections of the cells embedded in the resin but we would prefer to keep the coverslip in place if at all possible).
A parallel run of cells grown on Thermanox coverslips, processed at room temperature and embedded in conventional epoxy resin are working fine. I can easily get good ultrathin cross sections from these which includes the coverslip and cells.
My question is:
Has anyone had success with ultrathin cross sections of Thermanox coverslips and cells embedded in Lowicryl resins?
If so, any tricks or tips?
Many thanks, regards
Allan
Allan Mitchell Technical Manager Otago Centre for Electron Microscopy Department of Anatomy and Structural Biology School of Medical Sciences P.O. Box 913 Dunedin New Zealand
Phone (03) 479 5642 or 479 7301 Fax (03) 479 5086 or 479 7254
==============================Original Headers============================== 18, 20 -- From allan.mitchell-at-stonebow.otago.ac.nz Thu May 17 19:53:36 2007 18, 20 -- Received: from mailhub2.otago.ac.nz (mailhub2.otago.ac.nz [139.80.64.247]) 18, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4I0rZMW020878 18, 20 -- for {microscopy-at-msa.microscopy.com} ; Thu, 17 May 2007 19:53:35 -0500 18, 20 -- Received: from galadriel.otago.ac.nz (galadriel.otago.ac.nz [139.80.64.213]) 18, 20 -- by mailhub2.otago.ac.nz (8.13.8/8.13.8) with ESMTP id l4I0rOtw016069 18, 20 -- for {microscopy-at-msa.microscopy.com} ; Fri, 18 May 2007 12:53:33 +1200 18, 20 -- Received: from allan.otago.ac.nz ([139.80.40.92]) 18, 20 -- by galadriel.otago.ac.nz with esmtp (Exim 4.50) 18, 20 -- id 1HoqWh-0007Ht-Lg 18, 20 -- for microscopy-at-msa.microscopy.com; Fri, 18 May 2007 12:40:59 +1200 18, 20 -- Mime-Version: 1.0 (Apple Message framework v752.2) 18, 20 -- Content-Transfer-Encoding: 7bit 18, 20 -- Message-Id: {CF50D0A5-E4AB-49A6-AC84-B70F527A74B9-at-stonebow.otago.ac.nz} 18, 20 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 18, 20 -- To: microscopy-at-msa.microscopy.com 18, 20 -- From: Allan Mitchell {allan.mitchell-at-stonebow.otago.ac.nz} 18, 20 -- Subject: Cultured cells, thermanox coverslips and the PLT technique 18, 20 -- Date: Fri, 18 May 2007 12:40:58 +1200 18, 20 -- X-Mailer: Apple Mail (2.752.2) ==============================End of - Headers==============================
These sound like the "Top Hat" condenser apertures (since that is what they look like in cross section). They have a ~250micron "brim" which fits in the slots in the regular aperture holder rod and are ~3mm tall in the center. As with most apertures they have a tapered hole with the small end at the top. The purpose is to reduce the hard X-ray contribution (hole count) to the EDS spectrum. On FEG instruments with an adjustable C1 aperture, you can get essentially the same effect by using the C1 aperture as your beam limiter and using a C2 aperture slightly larger than the beam to stop the brehmstrahlung.
$8000 sounds ridiculously high for an aperture, even Platinum Top Hats. I got a package from FEI (p/n 9432 061 67001 for the 70micron) for ~$800. I don't remember offhand the number of pieces per package.
Henk
At 07:05 PM 5/17/2007, you wrote:
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==============================Original Headers============================== 10, 26 -- From colijn.1-at-osu.edu Thu May 17 19:58:07 2007 10, 26 -- Received: from ER6S1.ENG.OHIO-STATE.EDU (er6s1.ecr6.ohio-state.edu [164.107.76.2]) 10, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4I0w6Eh031712 10, 26 -- for {microscopy-at-microscopy.com} ; Thu, 17 May 2007 19:58:07 -0500 10, 26 -- Received: from CONVERSION-DAEMON.er6s1.eng.ohio-state.edu by 10, 26 -- er6s1.eng.ohio-state.edu (PMDF V6.2-1x11 #31056) 10, 26 -- id {01MGPFLZBYK0AA1TPQ-at-er6s1.eng.ohio-state.edu} for 10, 26 -- microscopy-at-microscopy.com; Thu, 17 May 2007 20:58:06 -0400 (EDT) 10, 26 -- Received: from HOC3.osu.edu 10, 26 -- (d149-67-33-174.col.wideopenwest.com [67.149.174.33]) 10, 26 -- by er6s1.eng.ohio-state.edu (PMDF V6.2-1x11 #31056) 10, 26 -- with ESMTPA id {01MGPFLY1E2MAAP4UD-at-er6s1.eng.ohio-state.edu} ; Thu, 10, 26 -- 17 May 2007 20:58:05 -0400 (EDT) 10, 26 -- Date: Thu, 17 May 2007 20:57:59 -0400 10, 26 -- From: "Hendrik O. Colijn" {colijn.1-at-osu.edu} 10, 26 -- Subject: Re: [Microscopy] C2 apertures 10, 26 -- In-reply-to: {200705172305.l4HN5u9b030905-at-ns.microscopy.com} 10, 26 -- Sender: colijn-at-er6s1.ecr6.ohio-state.edu 10, 26 -- To: john.mardinly-at-intel.com 10, 26 -- Cc: Microscopy Listserver {microscopy-at-microscopy.com} 10, 26 -- Message-id: {7.0.1.0.2.20070517204654.0037cc80-at-osu.edu} 10, 26 -- MIME-version: 1.0 10, 26 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 10, 26 -- Content-type: text/plain; charset=us-ascii; format=flowed 10, 26 -- X-Env-From: auth/colijn.1-at-osu.edu 10, 26 -- References: {200705172305.l4HN5u9b030905-at-ns.microscopy.com} ==============================End of - Headers==============================
Contact John Bishop -at- Norsam Technologies. http://www.norsam.com/ I talked to him in 2006 about a custom aperture strip for our Hitachi FIB. He may be able to make something for you but I don't know that it would be any cheaper.
Owen Mills Michigan Tech Univ.
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Krassimir; } 5-6 mm height? I would suspect that the actual aperture must be } inserted in some kind of holder, because it would seem to be difficult } to form an aperture from solid material that thick. The Pella web site } shows cross-sections of Philips apertures that are 3 mm diameter, and } 0.25 mm high, with a taper so that the thickness where the beam goes is } much less. However, I don't see anything 5-6 mm high in any catalogs, so } if the apertures are one piece, you might be stuck with the FEI deal. } } John Mardinly } Intel } } This is not an opinion of Intel Corp. } } -----Original Message----- } X-from: KN Bozhilov [mailto:bozhilov-at-ucr.edu] } Sent: Thursday, May 17, 2007 10:08 AM } To: microscopy-at-microscopy.com } Cc: Mardinly, John } Subject: Re: [Microscopy] RE: C2 apertures } } These are not disk foils. They look like a small cylinder with 3 mm } diameter and about 5-6 mm height. } } _______________________________________ } Krassimir N. Bozhilov } Central Facility for Advanced Microscopy and Microanalysis } University of California } Riverside, CA 92521 } } tel 951 827 2998 } fax 951 827 2489 } bozhilov-at-ucr.edu } _______________________________________ } } } On May 17, 2007, at 9:51 AM, john.mardinly-at-intel.com wrote: } } } } } } } } } ---------------------------------------------------------------------- } } } ------ } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } } America } } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } } MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------- } } } ------ } } } } Krassimir; } } That sounds like typical prices for FIB cut apertures for field } } emission TEMs, but is enough to make anyone say 'ouch!' I bought some } } cheap ones from Pella (~$60 as I recall) a little smaller than what I } } needed, and FIB cut them myself. If you have a FIB, it is pretty easy. } } } } John Mardinly } } Intel } } } } This is not an opinion of Intel Corporation. } } } } -----Original Message----- } } X-from: bozhilov-at-ucr.edu [mailto:bozhilov-at-ucr.edu] } } Sent: Thursday, May 17, 2007 9:36 AM } } To%3
==============================Original Headers============================== 7, 39 -- From opmills-at-mtu.edu Thu May 17 20:38:56 2007 7, 39 -- Received: from node7.mtu.edu (node7.mtu.edu [141.219.69.7]) 7, 39 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4I1cuk3012236 7, 39 -- for {Microscopy-at-microscopy.com} ; Thu, 17 May 2007 20:38:56 -0500 7, 39 -- Received: from node6.edge.dcsint.mtu.edu (node6.mtu.edu [141.219.69.6]) 7, 39 -- by node7.mtu.edu (8.13.1/8.13.1) with ESMTP id l4I1ctvu012427 7, 39 -- for {Microscopy-at-microscopy.com} ; Thu, 17 May 2007 21:38:55 -0400 7, 39 -- Received: from node34.edge.dcsint.mtu.edu (node34.mtu.edu [141.219.69.34]) 7, 39 -- by node6.edge.dcsint.mtu.edu (8.12.11.20060308/8.12.8) with ESMTP id l4I1ctAv027550 7, 39 -- for {Microscopy-at-microscopy.com} ; Thu, 17 May 2007 21:38:55 -0400 7, 39 -- Received: from mail.mtu.edu (campus0.mtu.edu [141.219.70.8]) 7, 39 -- by node34.edge.dcsint.mtu.edu (8.13.8/8.13.8) with ESMTP id l4I1ctAA007025 7, 39 -- for {Microscopy-at-microscopy.com} ; Thu, 17 May 2007 21:38:55 -0400 7, 39 -- (envelope-from opmills-at-mtu.edu) 7, 39 -- Received: from node5.edge.dcsint.mtu.edu (node5.mtu.edu [141.219.69.5]) 7, 39 -- by mail.mtu.edu (8.13.6+Sun/8.11.6) with ESMTP id l4I1csuE027420 7, 39 -- for {Microscopy-at-microscopy.com} ; Thu, 17 May 2007 21:38:54 -0400 (EDT) 7, 39 -- Received: from huskymail.mtu.edu (node75.mtu.edu [141.219.69.75]) 7, 39 -- by node5.edge.dcsint.mtu.edu (8.12.11.20060308/8.12.3/auth_ssl.mc v1.0) with ESMTP id l4I1cs4T015615 7, 39 -- for {Microscopy-at-microscopy.com} ; Thu, 17 May 2007 21:38:54 -0400 7, 39 -- Received: from 24.236.189.168 7, 39 -- (SquirrelMail authenticated user opmills) 7, 39 -- by huskymail.mtu.edu with HTTP; 7, 39 -- Thu, 17 May 2007 21:38:54 -0400 (EDT) 7, 39 -- Message-ID: {36388.24.236.189.168.1179452334.squirrel-at-huskymail.mtu.edu} 7, 39 -- In-Reply-To: {200705180058.l4I0wYIo000853-at-ns.microscopy.com} 7, 39 -- References: {200705180058.l4I0wYIo000853-at-ns.microscopy.com} 7, 39 -- Date: Thu, 17 May 2007 21:38:54 -0400 (EDT) 7, 39 -- Subject: Re: [Microscopy] C2 apertures 7, 39 -- From: opmills-at-mtu.edu 7, 39 -- To: Microscopy-at-microscopy.com 7, 39 -- User-Agent: SquirrelMail/1.4.8 7, 39 -- MIME-Version: 1.0 7, 39 -- Content-Type: text/plain;charset=iso-8859-1 7, 39 -- Content-Transfer-Encoding: 8bit 7, 39 -- X-Priority: 3 (Normal) 7, 39 -- Importance: Normal 7, 39 -- X-PMX-Version: 5.3.1.294258, Antispam-Engine: 2.5.0.283055, Antispam-Data: 2007.4.7.54434 7, 39 -- X-PerlMx-Spam: Gauge=X, Probability=10%, Report='PRIORITY_NO_NAME 0.716, NO_REAL_NAME 0, __C230066_P5 0, __CP_URI_IN_BODY 0, __CT 0, __CTE 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __HAS_X_PRIORITY 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0, __STOCK_PHRASE_7 0, __USER_AGENT 0' ==============================End of - Headers==============================
I agree with Henk here, that it is probably a "Top Hat" aperture. But these are typically about 1 mm in height (and hard to get a hold of for a JEOL 2000FX). My question to the group is, isn't there a hard, fixed aperture below the C2 aperture that is in the column and thicker like the one he is descibing?
-Scott
Scott D. Walck, Ph.D. Technical Director South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
US Toll Free: 1-800-728-2233 Tel: (949) 492-2600 Fax: (949) 492-1499
www.southbaytech.com walck-at-southbaytech.com
-----Original Message----- X-from: colijn.1-at-osu.edu [mailto:colijn.1-at-osu.edu] Sent: Thursday, May 17, 2007 6:01 PM To: Walck-at-SouthBayTech.com
Hi Krassimir and John,
These sound like the "Top Hat" condenser apertures (since that is what they look like in cross section). They have a ~250micron "brim" which fits in the slots in the regular aperture holder rod and are ~3mm tall in the center. As with most apertures they have a tapered hole with the small end at the top. The purpose is to reduce the hard X-ray contribution (hole count) to the EDS spectrum. On FEG instruments with an adjustable C1 aperture, you can get essentially the same effect by using the C1 aperture as your beam limiter and using a C2 aperture slightly larger than the beam to stop the brehmstrahlung.
$8000 sounds ridiculously high for an aperture, even Platinum Top Hats. I got a package from FEI (p/n 9432 061 67001 for the 70micron) for ~$800. I don't remember offhand the number of pieces per package.
Henk
At 07:05 PM 5/17/2007, you wrote:
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==============================Original Headers============================== 10, 26 -- From colijn.1-at-osu.edu Thu May 17 19:58:07 2007 10, 26 -- Received: from ER6S1.ENG.OHIO-STATE.EDU (er6s1.ecr6.ohio-state.edu [164.107.76.2]) 10, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4I0w6Eh031712 10, 26 -- for {microscopy-at-microscopy.com} ; Thu, 17 May 2007 19:58:07 -0500 10, 26 -- Received: from CONVERSION-DAEMON.er6s1.eng.ohio-state.edu by 10, 26 -- er6s1.eng.ohio-state.edu (PMDF V6.2-1x11 #31056) 10, 26 -- id {01MGPFLZBYK0AA1TPQ-at-er6s1.eng.ohio-state.edu} for 10, 26 -- microscopy-at-microscopy.com; Thu, 17 May 2007 20:58:06 -0400 (EDT) 10, 26 -- Received: from HOC3.osu.edu 10, 26 -- (d149-67-33-174.col.wideopenwest.com [67.149.174.33]) 10, 26 -- by er6s1.eng.ohio-state.edu (PMDF V6.2-1x11 #31056) 10, 26 -- with ESMTPA id {01MGPFLY1E2MAAP4UD-at-er6s1.eng.ohio-state.edu} ; Thu, 10, 26 -- 17 May 2007 20:58:05 -0400 (EDT) 10, 26 -- Date: Thu, 17 May 2007 20:57:59 -0400 10, 26 -- From: "Hendrik O. Colijn" {colijn.1-at-osu.edu} 10, 26 -- Subject: Re: [Microscopy] C2 apertures 10, 26 -- In-reply-to: {200705172305.l4HN5u9b030905-at-ns.microscopy.com} 10, 26 -- Sender: colijn-at-er6s1.ecr6.ohio-state.edu 10, 26 -- To: john.mardinly-at-intel.com 10, 26 -- Cc: Microscopy Listserver {microscopy-at-microscopy.com} 10, 26 -- Message-id: {7.0.1.0.2.20070517204654.0037cc80-at-osu.edu} 10, 26 -- MIME-version: 1.0 10, 26 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 10, 26 -- Content-type: text/plain; charset=us-ascii; format=flowed 10, 26 -- X-Env-From: auth/colijn.1-at-osu.edu 10, 26 -- References: {200705172305.l4HN5u9b030905-at-ns.microscopy.com} ==============================End of - Headers==============================
==============================Original Headers============================== 19, 21 -- From walck-at-southbaytech.com Thu May 17 22:45:14 2007 19, 21 -- Received: from flpvm23.prodigy.net (flpvm23.prodigy.net [207.115.20.53]) 19, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4I3jD7V025675 19, 21 -- for {Microscopy-at-microscopy.com} ; Thu, 17 May 2007 22:45:13 -0500 19, 21 -- X-ORBL: [64.169.217.123] 19, 21 -- Received: from dynamicbl8uno3 (adsl-64-169-217-123.dsl.lsan03.pacbell.net [64.169.217.123]) 19, 21 -- by flpvm23.prodigy.net (8.13.8 out.dk.spool/8.13.8) with ESMTP id l4I3jNEK032553 19, 21 -- for {Microscopy-at-microscopy.com} ; Thu, 17 May 2007 20:45:23 -0700 19, 21 -- From: "Scott Walck" {walck-at-southbaytech.com} 19, 21 -- To: {Microscopy-at-microscopy.com} 19, 21 -- Subject: RE: [Microscopy] Re: C2 apertures 19, 21 -- Date: Thu, 17 May 2007 20:45:39 -0700 19, 21 -- Message-ID: {004501c798ff$010c1540$7801a8c0-at-dynamicbl8uno3} 19, 21 -- MIME-Version: 1.0 19, 21 -- Content-Type: text/plain; 19, 21 -- charset="US-ASCII" 19, 21 -- Content-Transfer-Encoding: 7bit 19, 21 -- X-Mailer: Microsoft Office Outlook 11 19, 21 -- In-Reply-To: {200705180100.l4I10jMW005753-at-ns.microscopy.com} 19, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 19, 21 -- Thread-Index: AceY5/gPj1VTNd0cR1S3iDOLy0KoSgAFpArg ==============================End of - Headers==============================
In some of the older JEOL scopes (e.g. 200CX, 2000EX??) with a conventional (non C-O) objective lens you could get an optional HAX kit. This fit in the large upper bore of the pole-piece to reduce the brehmstrahlung contribution. In a scope with a condenser-objective, there is no room to put an additional absorber. There are no apertures between the C2 and the sample unless you want to call the upper bore of the objective an "aperture". {...grin...}
Krassimir was looking for an aperture for a CM300. There aren't any apertures between C2 and the sample.
Cheers, Henk
At 11:47 PM 05/17/07, you wrote:
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==============================Original Headers============================== 10, 26 -- From colijn.1-at-osu.edu Fri May 18 07:19:07 2007 10, 26 -- Received: from ER6S1.ENG.OHIO-STATE.EDU (er6s1.ecr6.ohio-state.edu [164.107.76.2]) 10, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4ICJ6gb015980 10, 26 -- for {microscopy-at-microscopy.com} ; Fri, 18 May 2007 07:19:06 -0500 10, 26 -- Received: from CONVERSION-DAEMON.er6s1.eng.ohio-state.edu by 10, 26 -- er6s1.eng.ohio-state.edu (PMDF V6.2-1x11 #31056) 10, 26 -- id {01MGQ3EAU8Y8A9UO2K-at-er6s1.eng.ohio-state.edu} for 10, 26 -- microscopy-at-microscopy.com; Fri, 18 May 2007 08:19:06 -0400 (EDT) 10, 26 -- Received: from HOC1.ecr6.ohio-state.edu 10, 26 -- (hoc1.ceof.ohio-state.edu [164.107.76.179]) by er6s1.eng.ohio-state.edu 10, 26 -- (PMDF V6.2-1x11 #31056) 10, 26 -- with ESMTPA id {01MGQ3EAHJ7UAAR18Q-at-er6s1.eng.ohio-state.edu} ; Fri, 10, 26 -- 18 May 2007 08:19:06 -0400 (EDT) 10, 26 -- Date: Fri, 18 May 2007 08:19:42 -0400 10, 26 -- From: "Hendrik O. Colijn" {colijn.1-at-osu.edu} 10, 26 -- Subject: Re: [Microscopy] C2 apertures 10, 26 -- In-reply-to: {200705180347.l4I3lDdv028296-at-ns.microscopy.com} 10, 26 -- Sender: colijn-at-er6s1.ecr6.ohio-state.edu 10, 26 -- To: walck-at-southbaytech.com 10, 26 -- Cc: Microscopy Listserver {microscopy-at-microscopy.com} 10, 26 -- Message-id: {7.0.1.0.2.20070518080457.0380fdb8-at-osu.edu} 10, 26 -- MIME-version: 1.0 10, 26 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 10, 26 -- Content-type: text/plain; charset=us-ascii; format=flowed 10, 26 -- X-Env-From: auth/colijn.1-at-osu.edu 10, 26 -- References: {200705180347.l4I3lDdv028296-at-ns.microscopy.com} ==============================End of - Headers==============================
With all due respect, making an O-ring is not a particularly difficult adventure. In fact, the very reference you are citing below has a kit Zatkoff sells to make your own custom O-rings.
(I don't know if one has to register with Zatkoff to see that link, but the registration is simple and quick if one is interested in looking into this.)
I certainly agree that it is better to just buy the proper O-ring, but sometimes the size you need is just not available easily without having to buy far more than you could need in the foreseeable future. I have had numerous occassions where just making the specific sized O-ring needed is just the simplest, most direct way to go.
dj
On Thu, 17 May 2007, bigelow-at-umich.edu wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } The Zatkoff Company (www.zatkoff.com) also carries an extensive stock } of O-rings. Before trying to make an O-ring yourself I would } recommend checking with them to see if they couldn't furnish } something to meet your needs. } -- } Wilbur C. Bigelow, Professor Emeritus } Materials Sci. & Engr., Univ. of Michigan } Ann Arbor, Michigan 48109-2136 } e-mail: bigelow-at-umich.edu; } Fx:734-763-4788; Ph:734-975-0858 } Address mail to: 2911 Whittier Court } Ann Arbor, MI 48104-6731 } } ==============================Original Headers============================== } 1, 14 -- From bigelow-at-umich.edu Thu May 17 14:37:50 2007 } 1, 14 -- Received: from skycaptain.mr.itd.umich.edu (smtp.mail.umich.edu [141.211.93.160]) } 1, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4HJbojt001489 } 1, 14 -- for {microscopy-at-microscopy.com} ; Thu, 17 May 2007 14:37:50 -0500 } 1, 14 -- Received: FROM [141.212.131.221] (bigelow-g4.engin.umich.edu [141.212.131.221]) } 1, 14 -- BY skycaptain.mr.itd.umich.edu ID 464CAF0D.60D1B.9874 ; } 1, 14 -- 17 May 2007 15:37:49 -0400 } 1, 14 -- Mime-Version: 1.0 } 1, 14 -- Message-Id: {p06240801c2725ef43d43-at-[141.212.131.221]} } 1, 14 -- Date: Thu, 17 May 2007 15:37:47 -0400 } 1, 14 -- To: Microscopy Listserver {microscopy-at-microscopy.com} } 1, 14 -- From: Wil Bigelow {bigelow-at-umich.edu} } 1, 14 -- Subject: [Microscopy}RE: O-ring } 1, 14 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" } ==============================End of - Headers============================== }
==============================Original Headers============================== 9, 19 -- From dljones-at-bestweb.net Fri May 18 07:55:47 2007 9, 19 -- Received: from vms044pub.verizon.net (vms044pub.verizon.net [206.46.252.44]) 9, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4ICtlq3028006 9, 19 -- for {Microscopy-at-microscopy.com} ; Fri, 18 May 2007 07:55:47 -0500 9, 19 -- Received: from localhost ([71.247.243.101]) 9, 19 -- by vms044.mailsrvcs.net (Sun Java System Messaging Server 6.2-6.01 (built Apr 9, 19 -- 3 2006)) with ESMTPA id {0JI8008M7MKBY727-at-vms044.mailsrvcs.net} for 9, 19 -- Microscopy-at-microscopy.com; Fri, 18 May 2007 07:55:29 -0500 (CDT) 9, 19 -- Date: Fri, 18 May 2007 08:56:19 -0400 (Eastern Daylight Time) 9, 19 -- From: "David L. Jones" {dljones-at-bestweb.net} 9, 19 -- Subject: Re: [Microscopy] [Microscopy}RE: O-ring 9, 19 -- In-reply-to: {200705171942.l4HJgrad007713-at-ns.microscopy.com} 9, 19 -- X-X-Sender: dljones-at-pop-croton.bestweb.net 9, 19 -- To: bigelow-at-umich.edu 9, 19 -- Cc: Microscopy-at-microscopy.com 9, 19 -- Message-id: {Pine.WNT.4.64.0705180846320.3096-at-H-F1} 9, 19 -- MIME-version: 1.0 9, 19 -- Content-type: TEXT/PLAIN; charset=US-ASCII; format=flowed 9, 19 -- References: {200705171942.l4HJgrad007713-at-ns.microscopy.com} ==============================End of - Headers==============================
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Organization: University of Zaragoza
Title-Subject: [Filtered] TEM and RuO4
Question: Dear Microscopist, my question is about TEM and the staining with RuO4. I am using this method to enhance the contrast of my sample and it is working properly, but I am trying to find some bibliography or some explanation about the mechanism, it is not very well described in any of the papers and book I could have a look. I know it is an oxydation proccess but how it is attaching to the molecule and more important to me, how it is modificating the size!!
Thank you for your help in advance! Kind Regards, patricia
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Email: thatch-at-mit.edu Name: Tori Hatch
Organization: HHMI
Title-Subject: [Filtered] automatic grid stainer
Question: I need to replace my Leica EM stainer. Leica is coming out with a new machine soon, but I'd like to hear from anyone who has experience with other automatic grid stainers.
In that case I would suggest to ask Apple. Contact them and tell them that for reasons of scientific Ethics you need the images in TIFF format (or any other format that you think might work), and ask them if they have a converter for their iMovie Product. They claim to be super user-friendly. Put them to the test.
mike
Michael Bode, Ph.D.
General Manager
OLYMPUS SOFT IMAGING SOLUTIONS 12596 West Bayaud Ave #300 Lakewood, CO 80228 USA Tel.: +1 (303) 234-9270 Fax.: +1 (303) 234-9271 E-mail: Mike.Bode-at-olympus-sis.com www.olympus-sis.com
-----Original Message----- X-from: Sharon Bledsoe [mailto:sbledsoe-at-iupui.edu] Sent: Friday, May 18, 2007 06:12 To: Mike Bode
} to replace PICT by PDF. This means that the use of PICT for exchanging } data will slowly diminish. } } It looks like sooner or later you will have to find a different choice } anyway. I would look at the whole chain of acquisition to see if you } cannot switch to something else. } } Why can't you use anything else? Does it have to do with the original } file format that can only be read by iMovie? Or are you tied to the Mac
} platform? } } mike } } } Michael Bode, Ph.D. } } General Manager } } OLYMPUS SOFT IMAGING SOLUTIONS } 12596 West Bayaud Ave #300 } Lakewood, CO 80228 } USA } Tel.: +1 (303) 234-9270 } Fax.: +1 (303) 234-9271 } E-mail: Mike.Bode-at-olympus-sis.com } www.olympus-sis.com } } -----Original Message----- } X-from: sbledsoe-at-iupui.edu [mailto:sbledsoe-at-iupui.edu] } Sent: Wednesday, May 16, 2007 19:39 } To: Mike Bode } Subject: [Microscopy] viaWWW: Need Help From Imaging Experts } } } } } ----------------------------------------------------------------------- } - } ---- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver
==============================Original Headers============================== 16, 25 -- From Mike.Bode-at-olympus-sis.com Fri May 18 09:59:35 2007 16, 25 -- Received: from mail.soft-imaging.de (ns.soft-imaging.de [62.180.61.130]) 16, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4IExYvC032651 16, 25 -- for {Microscopy-at-microscopy.com} ; Fri, 18 May 2007 09:59:35 -0500 16, 25 -- Received: from muenster.olympus-sis.com (muenster.olympus-sis.com [10.26.20.9]) 16, 25 -- by mail.soft-imaging.de (8.13.3/8.13.3/SuSE Linux 0.7) with ESMTP id l4IExcWK004449 16, 25 -- for {Microscopy-at-microscopy.com} ; Fri, 18 May 2007 16:59:40 +0200 16, 25 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 16, 25 -- Content-class: urn:content-classes:message 16, 25 -- MIME-Version: 1.0 16, 25 -- Content-Type: text/plain; 16, 25 -- charset="us-ascii" 16, 25 -- Subject: RE: [Microscopy] RE: viaWWW: Need Help From Imaging Experts 16, 25 -- Date: Fri, 18 May 2007 16:56:18 +0200 16, 25 -- Message-ID: {78B53BA1C5A2D9449EDA30A98800EC9493964F-at-ms-s-gws.soft-imaging.net} 16, 25 -- In-Reply-To: {a06240800c27347fa66ce-at-[134.68.251.140]} 16, 25 -- X-MS-Has-Attach: 16, 25 -- X-MS-TNEF-Correlator: 16, 25 -- Thread-Topic: [Microscopy] RE: viaWWW: Need Help From Imaging Experts 16, 25 -- Thread-Index: AceZRcBOo29V+b3ORW6do0b/i3cmNwAFostQ 16, 25 -- References: {200705171631.l4HGVqWh015155-at-ns.microscopy.com} {a06240800c27347fa66ce-at-[134.68.251.140]} 16, 25 -- From: "Mike Bode" {Mike.Bode-at-olympus-sis.com} 16, 25 -- To: {Microscopy-at-microscopy.com} 16, 25 -- Content-Transfer-Encoding: 8bit 16, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4IExYvC032651 ==============================End of - Headers==============================
The best reference I know is Trent, J.S. et al, Macromolecules, 1983, 16, 589. Trent deals with a variety of issues including specificity of staining by RuO4 and OsO4 and the chemistry of the reactions.
I suggest you also look in the Polymer Microscopy, by Sawyer and Grubbs, 2nd ed for other references.
Regards.
Disclaimer: The comments and opinions are those of the author alone and do not reflect any official position by Exxon Mobil Chemical Company.
Gary M. Brown Microscopy Specialist ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: 281 834 2387 fax: 281 834 2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
"Happiness equals reality minus expectations." Tom Magliozzi
pforcen-at-unizar .es To gary.m.brown-at-exxonmobil.com 05/18/07 08:29 cc AM Subject [Microscopy] viaWWW: TEM and RuO4 Please respond to pforcen-at-unizar .es
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Email: pforcen-at-unizar.es Name: Patricia
Organization: University of Zaragoza
Title-Subject: [Filtered] TEM and RuO4
Question: Dear Microscopist, my question is about TEM and the staining with RuO4. I am using this method to enhance the contrast of my sample and it is working properly, but I am trying to find some bibliography or some explanation about the mechanism, it is not very well described in any of the papers and book I could have a look. I know it is an oxydation proccess but how it is attaching to the molecule and more important to me, how it is modificating the size!!
Thank you for your help in advance! Kind Regards, patricia
I see what he means , just tried iMovie on OS X 10.3.
It seems to me though, that about 5 years ago, it DID let me save to TIFF and there were more options, perhaps trying to get an Old copy of iMovie? or try on an old machine if you still have OS 9?
Also can you open the clip in Quicktime Pro? Maybe Quicktime Pro will let you pull a tiff off of it, it too will save a frame.
Not a great solution, but if you are in a pinch.....
Lou Ann
Mike.Bode-at-olympus-sis.com wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } } In that case I would suggest to ask Apple. Contact them and tell them } that for reasons of scientific Ethics you need the images in TIFF format } (or any other format that you think might work), and ask them if they } have a converter for their iMovie Product. They claim to be super } user-friendly. Put them to the test. } } mike } } Michael Bode, Ph.D. } } General Manager } } OLYMPUS SOFT IMAGING SOLUTIONS } 12596 West Bayaud Ave #300 } Lakewood, CO 80228 } USA } Tel.: +1 (303) 234-9270 } Fax.: +1 (303) 234-9271 } E-mail: Mike.Bode-at-olympus-sis.com } www.olympus-sis.com } } -----Original Message----- } X-from: Sharon Bledsoe [mailto:sbledsoe-at-iupui.edu] } Sent: Friday, May 18, 2007 06:12 } To: Mike Bode } Subject: [Microscopy] RE: viaWWW: Need Help From Imaging Experts } } IF I had a choice I would choose to save my images as TIFF's, however, } the iMovie program will not let me. iMoive will only let me save as } PICT or JPEG. } } And I am not tied to the Mac, I'm chained to it. } } } } } } } ----------------------------------------------------------------------- } } ----- The Microscopy ListServer -- CoSponsor: The Microscopy Society } } of America To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ----------------------------------------------------------------------- } } ----- } } } } Hi Sharon, } } } } This may be a bit more fundamental than what you are looking for, but } } here goes: } } } } If you go to the MSA website, you can find a statement about ethical } } image processing that in essence says that images need to be stored } } uncompressed. JPEG is a format that includes lossy compression, and I } } believe that PICT can also contain lossy compression in the form of } } JPEG. Both therefore don't seem to be the best choices. } } } } I also found this on the web: } } } } In the next version of its operating system, MacOS X, Apple has decided } } } } } } to replace PICT by PDF. This means that the use of PICT for exchanging } } data will slowly diminish. } } } } It looks like sooner or later you will have to find a different choice } } anyway. I would look at the whole chain of acquisition to see if you } } cannot switch to something else. } } } } Why can't you use anything else? Does it have to do with the original } } file format that can only be read by iMovie? Or are you tied to the Mac } } } } } } platform? } } } } mike } } } } } } Michael Bode, Ph.D. } } } } General Manager } } } } OLYMPUS SOFT IMAGING SOLUTIONS } } 12596 West Bayaud Ave #300 } } Lakewood, CO 80228 } } USA } } Tel.: +1 (303) 234-9270 } } Fax.: +1 (303) 234-9271 } } E-mail: Mike.Bode-at-olympus-sis.com } } www.olympus-sis.com } } } } -----Original Message----- } } X-from: sbledsoe-at-iupui.edu [mailto:sbledsoe-at-iupui.edu] } } Sent: Wednesday, May 16, 2007 19:39 } } To: Mike Bode } } Subject: [Microscopy] viaWWW: Need Help From Imaging Experts } } } } } } } } } } ----------------------------------------------------------------------- } } - } } ---- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } } America To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ----------------------------------------------------------------------- } } - } } ---- } } } } This Question/Comment was submitted to the Microscopy Listserver using } } the WWW based Form at } } http://microscopy.com/MicroscopyListserver/MLFormMail.html } } ----------------------------------------------------------------------- } } - } } --- } } Remember this posting is most likely not from a Subscriber, so when } } replying } } please copy both sbledsoe-at-iupui.edu as well as the MIcroscopy } } Listserver } } ----------------------------------------------------------------------- } } - } } --- } } } } Email: sbledsoe-at-iupui.edu } } Name: Sharon Bledsoe } } } } Title-Subject: [Filtered] Need Help From Imaging Experts } } } } Question: Here is my problem: I can only use Mac iMovie to edit my } } digital video. I can not do this on a PC. ONLY iMovie. To save a } } single frame from the video, my choices are either PICT or JPEG format. } } } } } } My Question is: To save an origianl image, which is the best format, } } PICT or JPEG? } } } } I'm not interested in hearing about Tiffs, BMPs or any other format as } } that is not one of my choices. } } } } ----------------------------------------------------------------------- } } - } } --- } } } } ==============================Original } } Headers============================== } } 7, 11 -- From zaluzec-at-microscopy.com Wed May 16 20:28:23 2007 7, 11 -- } } Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) } } 7, 11 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } } ESMTP id l4H1SMmq032569 } } 7, 11 -- for {microscopy-at-microscopy.com} ; Wed, 16 May 2007 } } 20:28:22 -0500 } } 7, 11 -- Mime-Version: 1.0 } } 7, 11 -- Message-Id: {p06240801c27160281d64-at-[206.69.208.22]} } } 7, 11 -- Date: Wed, 16 May 2007 20:28:21 -0500 7, 11 -- To: } } microscopy-at-microscopy.com 7, 11 -- From: sbledsoe-at-iupui.edu (by way of } } MicroscopyListserver) 7, 11 -- Subject: viaWWW: Need Help From Imaging } } Experts 7, 11 -- Content-Type: text/plain; charset="us-ascii" } } ==============================End of - } } Headers============================== } } } } } } ==============================Original } } Headers============================== } } 26, 25 -- From Mike.Bode-at-olympus-sis.com Thu May 17 11:19:31 2007 26, } } 25 -- Received: from mail.soft-imaging.de (ns.soft-imaging.de } } [62.180.61.130]) } } 26, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } } ESMTP id l4HGJTqI003826 } } 26, 25 -- for {Microscopy-at-microscopy.com} ; Thu, 17 May 2007 } } 11:19:30 -0500 } } 26, 25 -- Received: from muenster.olympus-sis.com } } (muenster.olympus-sis.com [10.26.20.9]) } } 26, 25 -- by mail.soft-imaging.de (8.13.3/8.13.3/SuSE Linux } } 0.7) with ESMTP id l4HGJQd6014862 } } 26, 25 -- for {Microscopy-at-microscopy.com} ; Thu, 17 May 2007 } } 18:19:28 +0200 } } 26, 25 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 26, 25 -- } } Content-class: urn:content-classes:message 26, 25 -- MIME-Version: 1.0 } } 26, 25 -- Content-Type: text/plain; } } 26, 25 -- charset="us-ascii" } } 26, 25 -- Subject: RE: [Microscopy] viaWWW: Need Help From Imaging } } Experts 26, 25 -- Date: Thu, 17 May 2007 18:13:46 +0200 26, 25 -- } } Message-ID: } } {78B53BA1C5A2D9449EDA30A98800EC949394DD-at-ms-s-gws.soft-imaging.net} } } 26, 25 -- In-Reply-To: {200705170138.l4H1cq2c024253-at-ns.microscopy.com} } } 26, 25 -- X-MS-Has-Attach: } } 26, 25 -- X-MS-TNEF-Correlator: } } 26, 25 -- Thread-Topic: [Microscopy] viaWWW: Need Help From Imaging } } Experts 26, 25 -- Thread-Index: AceYJCKWCu3O+yiVSbqbAjFTBkbwPwAeNnzw } } 26, 25 -- References: {200705170138.l4H1cq2c024253-at-ns.microscopy.com} } } 26, 25 -- From: "Mike Bode" {Mike.Bode-at-olympus-sis.com} 26, 25 -- To: } } {Microscopy-at-microscopy.com} 26, 25 -- Content-Transfer-Encoding: 8bit } } 26, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } } ns.microscopy.com id l4HGJTqI003826 ==============================End } } of - Headers============================== } } } } } } ==============================Original Headers============================== } 16, 25 -- From Mike.Bode-at-olympus-sis.com Fri May 18 09:59:35 2007 } 16, 25 -- Received: from mail.soft-imaging.de (ns.soft-imaging.de [62.180.61.130]) } 16, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4IExYvC032651 } 16, 25 -- for {Microscopy-at-microscopy.com} ; Fri, 18 May 2007 09:59:35 -0500 } 16, 25 -- Received: from muenster.olympus-sis.com (muenster.olympus-sis.com [10.26.20.9]) } 16, 25 -- by mail.soft-imaging.de (8.13.3/8.13.3/SuSE Linux 0.7) with ESMTP id l4IExcWK004449 } 16, 25 -- for {Microscopy-at-microscopy.com} ; Fri, 18 May 2007 16:59:40 +0200 } 16, 25 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 } 16, 25 -- Content-class: urn:content-classes:message } 16, 25 -- MIME-Version: 1.0 } 16, 25 -- Content-Type: text/plain; } 16, 25 -- charset="us-ascii" } 16, 25 -- Subject: RE: [Microscopy] RE: viaWWW: Need Help From Imaging Experts } 16, 25 -- Date: Fri, 18 May 2007 16:56:18 +0200 } 16, 25 -- Message-ID: {78B53BA1C5A2D9449EDA30A98800EC9493964F-at-ms-s-gws.soft-imaging.net} } 16, 25 -- In-Reply-To: {a06240800c27347fa66ce-at-[134.68.251.140]} } 16, 25 -- X-MS-Has-Attach: } 16, 25 -- X-MS-TNEF-Correlator: } 16, 25 -- Thread-Topic: [Microscopy] RE: viaWWW: Need Help From Imaging Experts } 16, 25 -- Thread-Index: AceZRcBOo29V+b3ORW6do0b/i3cmNwAFostQ } 16, 25 -- References: {200705171631.l4HGVqWh015155-at-ns.microscopy.com} {a06240800c27347fa66ce-at-[134.68.251.140]} } 16, 25 -- From: "Mike Bode" {Mike.Bode-at-olympus-sis.com} } 16, 25 -- To: {Microscopy-at-microscopy.com} } 16, 25 -- Content-Transfer-Encoding: 8bit } 16, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4IExYvC032651 } ==============================End of - Headers============================== } }
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Lou Ann Miller, MT(ASCP) Service Supervisor Center for Microscopic Imaging College of Veterinary Medicine Rm 1204 VMBSB 2001 S Lincoln Ave Urbana, IL 61802
217/244-1567 http://treefrog.cvm.uiuc.edu
==============================Original Headers============================== 12, 19 -- From lamiller-at-uiuc.edu Fri May 18 10:17:26 2007 12, 19 -- Received: from expredir6.cites.uiuc.edu (expredir6.cites.uiuc.edu [128.174.5.97]) 12, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4IFHQUv019632 12, 19 -- for {microscopy-at-microscopy.com} ; Fri, 18 May 2007 10:17:26 -0500 12, 19 -- Received: from redkite.cvm.uiuc.edu (redkite.cvm.uiuc.edu [130.126.18.157]) 12, 19 -- by expredir6.cites.uiuc.edu (8.14.1/8.14.1) with ESMTP id l4IFHQu3027443 12, 19 -- for {microscopy-at-microscopy.com} ; Fri, 18 May 2007 10:17:26 -0500 (CDT) 12, 19 -- Message-ID: {464DC386.6010806-at-uiuc.edu} 12, 19 -- Date: Fri, 18 May 2007 10:17:26 -0500 12, 19 -- From: Lou Ann Miller {lamiller-at-uiuc.edu} 12, 19 -- Reply-To: lamiller-at-uiuc.edu 12, 19 -- User-Agent: Thunderbird 2.0.0.0 (Macintosh/20070326) 12, 19 -- MIME-Version: 1.0 12, 19 -- To: microscopy-at-microscopy.com 12, 19 -- Subject: Re: [Microscopy] viaWWW: Need Help From Imaging Experts 12, 19 -- References: {200705181500.l4IF0Scg001229-at-ns.microscopy.com} 12, 19 -- In-Reply-To: {200705181500.l4IF0Scg001229-at-ns.microscopy.com} 12, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 12, 19 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
We have had the same problem in that only the epoxy resins will bind to the thermanox, and aclar for that matter. Are you doing PLT and HM20 embedding for immunolabeling, or just to avoid room temp dehydration? I know some people who have had success immunolabeling after etching the epoxy resin away (just an idea). We have had the most success growing cells on a Transwell Plate (sold through Corning here in Canada) that has a porous polyester membrane on the bottom. The HM20 resin is able to penetrate through the pores and will hold everything together during sectioning allowing for decent thin sectioning. Expect some wrinkles and getting a perfect flat cross section is difficult because the membrane is pretty flexible, but it works. Why is it so important to have the coverslip present?
Hope this helps,
Garnet
} Hi all } } I am having some sectioning difficulties with cross sections of } cultured cells grown on Thermanox coverslips. } } The monolayer of cells grown on the coverslips have been processed by } the progressive lowering of temperature technique (in an AFS), using } ethanol as the dehydration solvent, and then followed by embedding in } Lowicryl HM20. } } The problem I am having is that the Thermanox coverslips are } separating from the Lowicryl HM20 resin block making cross-sectional } ultrathin sectioning (and semi-thin for that matter) that includes } the coverslip difficult. } } The separation is easily seen through the binocular head of the } ultramicrotome even before I start trimming the block down. The } resin simply hasn't 'bonded' to the coverslip. (I am able to } remove the coverslip from the block and then get good ultrathin } sections of the cells embedded in the resin but we would prefer to } keep the coverslip in place if at all possible). } } A parallel run of cells grown on Thermanox coverslips, processed at } room temperature and embedded in conventional epoxy resin are working } fine. I can easily get good ultrathin cross sections from these } which includes the coverslip and cells. } } My question is: } } Has anyone had success with ultrathin cross sections of Thermanox } coverslips and cells embedded in Lowicryl resins? } } If so, any tricks or tips? } } } Many thanks, regards } } Allan } } } } Allan Mitchell } Technical Manager } Otago Centre for Electron Microscopy } Department of Anatomy and Structural Biology } School of Medical Sciences } P.O. Box 913 } Dunedin } New Zealand } } Phone (03) 479 5642 or 479 7301 } Fax (03) 479 5086 or 479 7254 } } } } ==============================Original Headers============================== } 18, 20 -- From allan.mitchell-at-stonebow.otago.ac.nz Thu May 17 19:53:36 2007 } 18, 20 -- Received: from mailhub2.otago.ac.nz (mailhub2.otago.ac.nz } [139.80.64.247]) } 18, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id l4I0rZMW020878 } 18, 20 -- for {microscopy-at-msa.microscopy.com} ; Thu, 17 May 2007 } 19:53:35 -0500 } 18, 20 -- Received: from galadriel.otago.ac.nz } (galadriel.otago.ac.nz [139.80.64.213]) } 18, 20 -- by mailhub2.otago.ac.nz (8.13.8/8.13.8) with ESMTP id } l4I0rOtw016069 } 18, 20 -- for {microscopy-at-msa.microscopy.com} ; Fri, 18 May 2007 } 12:53:33 +1200 } 18, 20 -- Received: from allan.otago.ac.nz ([139.80.40.92]) } 18, 20 -- by galadriel.otago.ac.nz with esmtp (Exim 4.50) } 18, 20 -- id 1HoqWh-0007Ht-Lg } 18, 20 -- for microscopy-at-msa.microscopy.com; Fri, 18 May 2007 } 12:40:59 +1200 } 18, 20 -- Mime-Version: 1.0 (Apple Message framework v752.2) } 18, 20 -- Content-Transfer-Encoding: 7bit } 18, 20 -- Message-Id: } {CF50D0A5-E4AB-49A6-AC84-B70F527A74B9-at-stonebow.otago.ac.nz} } 18, 20 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed } 18, 20 -- To: microscopy-at-msa.microscopy.com } 18, 20 -- From: Allan Mitchell {allan.mitchell-at-stonebow.otago.ac.nz} } 18, 20 -- Subject: Cultured cells, thermanox coverslips and the PLT technique } 18, 20 -- Date: Fri, 18 May 2007 12:40:58 +1200 } 18, 20 -- X-Mailer: Apple Mail (2.752.2) } ==============================End of - Headers==============================
-- Garnet Martens
Research Manager BioImaging Facility University of British Columbia 6270 University Blvd. Vancouver, B.C. Canada V6T 1Z4
phone 604-822-3354
==============================Original Headers============================== 10, 31 -- From gmartens-at-interchange.ubc.ca Fri May 18 10:21:46 2007 10, 31 -- Received: from mr8.mail-relay.ubc.ca (mr8.mail-relay.ubc.ca [137.82.45.15]) 10, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4IFLiAx029426 10, 31 -- for {microscopy-at-msa.microscopy.com} ; Fri, 18 May 2007 10:21:46 -0500 10, 31 -- Received: from mr8.mail-relay.ubc.ca (localhost [127.0.0.1]) 10, 31 -- by localhost (Postfix) with SMTP id 3E5FD10B16 10, 31 -- for {microscopy-at-msa.microscopy.com} ; Fri, 18 May 2007 08:21:44 -0700 (PDT) 10, 31 -- Received: from mta2.interchange.ubc.ca (mta2.interchange.ubc.ca [142.103.145.70]) 10, 31 -- by mr8.mail-relay.ubc.ca (Postfix) with ESMTP 10, 31 -- for {microscopy-at-msa.microscopy.com} ; Fri, 18 May 2007 08:21:43 -0700 (PDT) 10, 31 -- Received: from [137.82.85.216] (0511.emlab.ubc.ca [137.82.85.216]) 10, 31 -- by smtp.interchange.ubc.ca 10, 31 -- (iPlanet Messaging Server 5.2 HotFix 1.21 (built Sep 8 2003)) 10, 31 -- with ESMTPA id {0JI8002EBTC69Z-at-smtp.interchange.ubc.ca} for 10, 31 -- microscopy-at-msa.microscopy.com; Fri, 18 May 2007 08:21:43 -0700 (PDT) 10, 31 -- Date: Fri, 18 May 2007 08:21:42 -0700 10, 31 -- From: Garnet Martens {gmartens-at-interchange.ubc.ca} 10, 31 -- Subject: Re: [Microscopy] Cultured cells, 10, 31 -- thermanox coverslips and the PLT technique 10, 31 -- In-reply-to: {200705180056.l4I0uE02026743-at-ns.microscopy.com} 10, 31 -- To: allan.mitchell-at-stonebow.otago.ac.nz 10, 31 -- Cc: microscopy-at-msa.microscopy.com 10, 31 -- Message-id: {a06240801c27372db7f35-at-[137.82.85.216]} 10, 31 -- MIME-version: 1.0 10, 31 -- Content-type: text/plain; format=flowed; charset=us-ascii 10, 31 -- References: {200705180056.l4I0uE02026743-at-ns.microscopy.com} 10, 31 -- X-UBC-Scanned: Sophos PureMessage 5.3.1.294258, Antispam-Engine: 2.5.0.283055, Antispam-Data: 2007.5.18.80041 10, 31 -- X-UBC-Relayed: Relayed through mail-relay.ubc.ca 10, 31 -- X-PerlMx-Spam: Probability=7%, Report=__C230066_P5 0, __CT 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0, __STOCK_PHRASE_24 0 10, 31 -- X-Spam-Level: 10, 31 -- X-Spam-Flag: No ==============================End of - Headers==============================
The PICT files captured through iMovie are lossless, so this would be the flavor of choice. You can then easily convert PICT to TIFF if you wish.
Cheers,
-- Tracy E. Anderson Microscopist and digital imaging specialist Imaging Center University of Minnesota College of Biological Sciences Phone: 612.624.3454 Fax: 612.624.1799 http://www.cbs.umn.edu/ic/ -- "Science and art belong to the whole world, and before them vanish the barriers of nationality." - Goethe
On 18 May 2007, Mike.Bode-at-olympus-sis.com wrote: } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } } In that case I would suggest to ask Apple. Contact them and tell them } that for reasons of scientific Ethics you need the images in TIFF format } (or any other format that you think might work), and ask them if they } have a converter for their iMovie Product. They claim to be super } user-friendly. Put them to the test. } } mike } } Michael Bode, Ph.D. } } General Manager } } OLYMPUS SOFT IMAGING SOLUTIONS } 12596 West Bayaud Ave #300 } Lakewood, CO 80228 } USA } Tel.: +1 (303) 234-9270 } Fax.: +1 (303) 234-9271 } E-mail: Mike.Bode-at-olympus-sis.com } www.olympus-sis.com } } -----Original Message----- } X-from: Sharon Bledsoe [mailto:sbledsoe-at-iupui.edu] } Sent: Friday, May 18, 2007 06:12 } To: Mike Bode } Subject: [Microscopy] RE: viaWWW: Need Help From Imaging Experts } } IF I had a choice I would choose to save my images as TIFF's, however, } the iMovie program will not let me. iMoive will only let me save as } PICT or JPEG. } } And I am not tied to the Mac, I'm chained to it. } } } } } } ----------------------------------------------------------------------- } } ----- The Microscopy ListServer -- CoSponsor: The Microscopy Society } } of America To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ----------------------------------------------------------------------- } } ----- } } } } Hi Sharon, } } } } This may be a bit more fundamental than what you are looking for, but } } here goes: } } } } If you go to the MSA website, you can find a statement about ethical } } image processing that in essence says that images need to be stored } } uncompressed. JPEG is a format that includes lossy compression, and I } } believe that PICT can also contain lossy compression in the form of } } JPEG. Both therefore don't seem to be the best choices. } } } } I also found this on the web: } } } } In the next version of its operating system, MacOS X, Apple has decided } } } to replace PICT by PDF. This means that the use of PICT for exchanging } } data will slowly diminish. } } } } It looks like sooner or later you will have to find a different choice } } anyway. I would look at the whole chain of acquisition to see if you } } cannot switch to something else. } } } } Why can't you use anything else? Does it have to do with the original } } file format that can only be read by iMovie? Or are you tied to the Mac } } } platform? } } } } mike } } } } } } Michael Bode, Ph.D. } } } } General Manager } } } } OLYMPUS SOFT IMAGING SOLUTIONS } } 12596 West Bayaud Ave #300 } } Lakewood, CO 80228 } } USA } } Tel.: +1 (303) 234-9270 } } Fax.: +1 (303) 234-9271 } } E-mail: Mike.Bode-at-olympus-sis.com } } www.olympus-sis.com } } } } -----Original Message----- } } X-from: sbledsoe-at-iupui.edu [mailto:sbledsoe-at-iupui.edu] } } Sent: Wednesday, May 16, 2007 19:39 } } To: Mike Bode } } Subject: [Microscopy] viaWWW: Need Help From Imaging Experts } } } } } } } } } } ----------------------------------------------------------------------- } } - } } ---- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } } America To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ----------------------------------------------------------------------- } } - } } ---- } } } } This Question/Comment was submitted to the Microscopy Listserver using } } the WWW based Form at } } http://microscopy.com/MicroscopyListserver/MLFormMail.html } } ----------------------------------------------------------------------- } } - } } --- } } Remember this posting is most likely not from a Subscriber, so when } } replying } } please copy both sbledsoe-at-iupui.edu as well as the MIcroscopy } } Listserver } } ----------------------------------------------------------------------- } } - } } --- } } } } Email: sbledsoe-at-iupui.edu } } Name: Sharon Bledsoe } } } } Title-Subject: [Filtered] Need Help From Imaging Experts } } } } Question: Here is my problem: I can only use Mac iMovie to edit my } } digital video. I can not do this on a PC. ONLY iMovie. To save a } } single frame from the video, my choices are either PICT or JPEG format. } } } } } } My Question is: To save an origianl image, which is the best format, } } PICT or JPEG? } } } } I'm not interested in hearing about Tiffs, BMPs or any other format as } } that is not one of my choices. } } } } ----------------------------------------------------------------------- } } - } } --- } } } } ==============================Original } } Headers============================== } } 7, 11 -- From zaluzec-at-microscopy.com Wed May 16 20:28:23 2007 7, 11 -- } } Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) } } 7, 11 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } } ESMTP id l4H1SMmq032569 } } 7, 11 -- for {microscopy-at-microscopy.com} ; Wed, 16 May 2007 } } 20:28:22 -0500 } } 7, 11 -- Mime-Version: 1.0 } } 7, 11 -- Message-Id: {p06240801c27160281d64-at-[206.69.208.22]} } } 7, 11 -- Date: Wed, 16 May 2007 20:28:21 -0500 7, 11 -- To: } } microscopy-at-microscopy.com 7, 11 -- From: sbledsoe-at-iupui.edu (by way of } } MicroscopyListserver) 7, 11 -- Subject: viaWWW: Need Help From Imaging } } Experts 7, 11 -- Content-Type: text/plain; charset="us-ascii" } } ==============================End of - } } Headers============================== } } } } } } ==============================Original } } Headers============================== } } 26, 25 -- From Mike.Bode-at-olympus-sis.com Thu May 17 11:19:31 2007 26, } } 25 -- Received: from mail.soft-imaging.de (ns.soft-imaging.de } } [62.180.61.130]) } } 26, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } } ESMTP id l4HGJTqI003826 } } 26, 25 -- for {Microscopy-at-microscopy.com} ; Thu, 17 May 2007 } } 11:19:30 -0500 } } 26, 25 -- Received: from muenster.olympus-sis.com } } (muenster.olympus-sis.com [10.26.20.9]) } } 26, 25 -- by mail.soft-imaging.de (8.13.3/8.13.3/SuSE Linux } } 0.7) with ESMTP id l4HGJQd6014862 } } 26, 25 -- for {Microscopy-at-microscopy.com} ; Thu, 17 May 2007 } } 18:19:28 +0200 } } 26, 25 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 26, 25 -- } } Content-class: urn:content-classes:message 26, 25 -- MIME-Version: 1.0 } } 26, 25 -- Content-Type: text/plain; } } 26, 25 -- charset="us-ascii" } } 26, 25 -- Subject: RE: [Microscopy] viaWWW: Need Help From Imaging } } Experts 26, 25 -- Date: Thu, 17 May 2007 18:13:46 +0200 26, 25 -- } } Message-ID: } } {78B53BA1C5A2D9449EDA30A98800EC949394DD-at-ms-s-gws.soft-imaging.net} } } 26, 25 -- In-Reply-To: {200705170138.l4H1cq2c024253-at-ns.microscopy.com} } } 26, 25 -- X-MS-Has-Attach: } } 26, 25 -- X-MS-TNEF-Correlator: } } 26, 25 -- Thread-Topic: [Microscopy] viaWWW: Need Help From Imaging } } Experts 26, 25 -- Thread-Index: AceYJCKWCu3O+yiVSbqbAjFTBkbwPwAeNnzw } } 26, 25 -- References: {200705170138.l4H1cq2c024253-at-ns.microscopy.com} } } 26, 25 -- From: "Mike Bode" {Mike.Bode-at-olympus-sis.com} 26, 25 -- To: } } {Microscopy-at-microscopy.com} 26, 25 -- Content-Transfer-Encoding: 8bit } } 26, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } } ns.microscopy.com id l4HGJTqI003826 ==============================End } } of - Headers============================== } } } } ==============================Original Headers============================== } 16, 25 -- From Mike.Bode-at-olympus-sis.com Fri May 18 09:59:35 2007 } 16, 25 -- Received: from mail.soft-imaging.de (ns.soft-imaging.de } [62.180.61.130]) } 16, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } l4IExYvC032651 } 16, 25 -- for {Microscopy-at-microscopy.com} ; Fri, 18 May 2007 09:59:35 -0500 } 16, 25 -- Received: from muenster.olympus-sis.com (muenster.olympus-sis.com } [10.26.20.9]) } 16, 25 -- by mail.soft-imaging.de (8.13.3/8.13.3/SuSE Linux 0.7) with ESMTP } id l4IExcWK004449 } 16, 25 -- for {Microscopy-at-microscopy.com} ; Fri, 18 May 2007 16:59:40 +0200 } 16, 25 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 } 16, 25 -- Content-class: urn:content-classes:message } 16, 25 -- MIME-Version: 1.0 } 16, 25 -- Content-Type: text/plain; } 16, 25 -- charset="us-ascii" } 16, 25 -- Subject: RE: [Microscopy] RE: viaWWW: Need Help From Imaging } Experts } 16, 25 -- Date: Fri, 18 May 2007 16:56:18 +0200 } 16, 25 -- Message-ID: } {78B53BA1C5A2D9449EDA30A98800EC9493964F-at-ms-s-gws.soft-imaging.net} } 16, 25 -- In-Reply-To: {a06240800c27347fa66ce-at-[134.68.251.140]} } 16, 25 -- X-MS-Has-Attach: } 16, 25 -- X-MS-TNEF-Correlator: } 16, 25 -- Thread-Topic: [Microscopy] RE: viaWWW: Need Help From Imaging } Experts } 16, 25 -- Thread-Index: AceZRcBOo29V+b3ORW6do0b/i3cmNwAFostQ } 16, 25 -- References: {200705171631.l4HGVqWh015155-at-ns.microscopy.com} } {a06240800c27347fa66ce-at-[134.68.251.140]} } 16, 25 -- From: "Mike Bode" {Mike.Bode-at-olympus-sis.com} } 16, 25 -- To: {Microscopy-at-microscopy.com} } 16, 25 -- Content-Transfer-Encoding: 8bit } 16, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } ns.microscopy.com id l4IExYvC032651 } ==============================End of - } Headers============================== }
==============================Original Headers============================== 10, 20 -- From trazy-at-umn.edu Fri May 18 10:26:14 2007 10, 20 -- Received: from mta-a2.tc.umn.edu (mta-a2.tc.umn.edu [134.84.119.206]) 10, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4IFQE8g006446 10, 20 -- for {microscopy-at-microscopy.com} ; Fri, 18 May 2007 10:26:14 -0500 10, 20 -- Received: from vanguard.software.umn.edu (vanguard.software.umn.edu [128.101.65.55]) 10, 20 -- by mta-a2.tc.umn.edu (UMN smtpd) with ESMTP 10, 20 -- Fri, 18 May 2007 10:26:07 -0500 (CDT) 10, 20 -- X-Umn-Remote-Mta: [N] vanguard.software.umn.edu [128.101.65.55] #+LO+TR 10, 20 -- Received: (from nobody-at-localhost) 10, 20 -- by vanguard.software.umn.edu (8.12.11/8.12.8) id l4IFQ7db017180; 10, 20 -- Fri, 18 May 2007 10:26:07 -0500 10, 20 -- Message-Id: {200705181526.l4IFQ7db017180-at-vanguard.software.umn.edu} 10, 20 -- Date: Fri, 18 May 2007 10:26:07 CDT 10, 20 -- From: tracy {trazy-at-umn.edu} 10, 20 -- Subject: Re: [Microscopy] viaWWW: Need Help From Imaging Experts 10, 20 -- To: Mike.Bode-at-olympus-sis.com, microscopy-at-microscopy.com 10, 20 -- MIME-Version: 1.0 10, 20 -- Content-Type: TEXT/plain; CHARSET=US-ASCII 10, 20 -- X-Tick-Nemesis: American Maid 10, 20 -- X-remote-user-ip: 160.94.80.149 ==============================End of - Headers==============================
These are straightforward C2 apertures and sit in the C2 aperture holder rod. They do have the function of the "top hat" apertures but they do not have the "top hat" shape. It is a simple cylinder with 3mm diameter and (after I measured again) 4 mm height. I do not know what is the material it is made of. I am surprised that these apertures turn out to be so esotheric objects, after all CM300 is not that of a surreptitious instrument.
Krassimir. _______________________________________ Krassimir N. Bozhilov Central Facility for Advanced Microscopy and Microanalysis University of California Riverside, CA 92521
tel 951 827 2998 fax 951 827 2489 bozhilov-at-ucr.edu _______________________________________
On May 17, 2007, at 8:47 PM, walck-at-southbaytech.com wrote:
} } } } ---------------------------------------------------------------------- } ------ } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } I agree with Henk here, that it is probably a "Top Hat" aperture. } But these } are typically about 1 mm in height (and hard to get a hold of for a } JEOL } 2000FX). My question to the group is, isn't there a hard, fixed } aperture } below the C2 aperture that is in the column and thicker like the } one he is } descibing? } } } -Scott } } Scott D. Walck, Ph.D. } Technical Director } South Bay Technology, Inc. } 1120 Via Callejon } San Clemente, CA 92673 } } US Toll Free: 1-800-728-2233 } Tel: (949) 492-2600 } Fax: (949) 492-1499 } } www.southbaytech.com } walck-at-southbaytech.com } } -----Original Message----- } X-from: colijn.1-at-osu.edu [mailto:colijn.1-at-osu.edu] } Sent: Thursday, May 17, 2007 6:01 PM } To: Walck-at-SouthBayTech.com } Subject: [Microscopy] Re: C2 apertures } } } } } ---------------------------------------------------------------------- } ------ } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } Hi Krassimir and John, } } These sound like the "Top Hat" condenser apertures (since that is } what they } look like in cross section). They have a ~250micron "brim" } which fits in the slots in the regular aperture holder rod and are } ~3mm tall } in the center. As with most apertures they have a tapered hole } with the } small end at the top. The purpose is to reduce the hard X-ray } contribution } (hole count) to the EDS spectrum. On FEG instruments with an } adjustable C1 } aperture, you can get essentially the same effect by using the C1 } aperture } as your beam limiter and using a C2 aperture slightly larger than } the beam } to stop the brehmstrahlung. } } $8000 sounds ridiculously high for an aperture, even Platinum Top } Hats. I } got a package from FEI (p/n 9432 061 67001 for the } 70micron) for ~$800. I don't remember offhand the number of } pieces per } package. } } Henk } } At 07:05 PM 5/17/2007, you wrote: } } } } } --------------------------------------------------------------------- } } -- } } ----- The Microscopy ListServer -- CoSponsor: The Microscopy Society } } of America To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } --------------------------------------------------------------------- } } -- } } ----- } } } } Krassimir; } } 5-6 mm height? I would suspect that the actual aperture must } } be inserted in some kind of holder, because it would seem to be } } difficult to form an aperture from solid material that thick. The } } Pella } } web site shows cross-sections of Philips apertures that are 3 mm } } diameter, and } } 0.25 mm high, with a taper so that the thickness where the beam } } goes is } } much less. However, I don't see anything 5-6 mm high in any catalogs, } } so if the apertures are one piece, you might be stuck with the FEI } } deal. } } } } John Mardinly } } Intel } } } } This is not an opinion of Intel Corp. } } } } -----Original Message----- } } X-from: KN Bozhilov [mailto:bozhilov-at-ucr.edu] } } Sent: Thursday, May 17, 2007 10:08 AM } } To: microscopy-at-microscopy.com } } Cc: Mardinly, John } } Subject: Re: [Microscopy] RE: C2 apertures } } } } These are not disk foils. They look like a small cylinder with 3 mm } } diameter and about 5-6 mm height. } } } } _______________________________________ } } Krassimir N. Bozhilov } } Central Facility for Advanced Microscopy and Microanalysis University } } of California Riverside, CA 92521 } } } } tel 951 827 2998 } } fax 951 827 2489 } } bozhilov-at-ucr.edu } } _______________________________________ } } } } } } On May 17, 2007, at 9:51 AM, john.mardinly-at-intel.com wrote: } } } } } } } } } } } } } } -------------------------------------------------------------------- } } } -- } } } } } ------ } } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } } } America To Subscribe/Unsubscribe -- http://www.microscopy.com/ } } } MicroscopyListserver On-Line Help } } } http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } -------------------------------------------------------------------- } } } -- } } } } } ------ } } } } } } Krassimir; } } } That sounds like typical prices for FIB cut apertures for } } } field emission TEMs, but is enough to make anyone say 'ouch!' I } } } bought some cheap ones from Pella (~$60 as I recall) a little } } } smaller than what I needed, and FIB cut them myself. If you have } } } a FIB, } it is pretty easy. } } } } } } John Mardinly } } } Intel } } } } } } This is not an opinion of Intel Corporation. } } } } } } -----Original Message----- } } } X-from: bozhilov-at-ucr.edu [mailto:bozhilov-at-ucr.edu] } } } Sent: Thursday, May 17, 2007 9:36 AM } } } To: Mardinly, John } } } Subject: [Microscopy] C2 apertures } } } } } } } } } } } } } } } -------------------------------------------------------------------- } } } -- } } } } } -- } } } ---- } } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } } } America To Subscribe/Unsubscribe -- } } } http://www.microscopy.com/MicroscopyListserver } } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } -------------------------------------------------------------------- } } } -- } } } } } -- } } } ---- } } } } } } Looking for second condenser (C2) low-background 15, 30, 50 and 100 } } } micron apertures for FEI CM300 TEM. } } } } } } These are not the standard Pt thin foil discs. } } } I got a quote for a set for approx. $8000 from FEI. Any alternative } } } suppliers out there? } } } } } } Thanks, } } } } } } Krassimir. } } } _______________________________________ } } } Krassimir N. Bozhilov } } } Central Facility for Advanced Microscopy and Microanalysis } } } University of California Riverside, CA 92521 } } } } } } tel 951 827 2998 } } } fax 951 827 2489 } } } bozhilov-at-ucr.edu } } } _______________________________________ } } } } } } } } } } } } ==============================Original } } } Headers============================== } } } 7, 19 -- From bozhilov-at-ucr.edu Thu May 17 11:35:52 2007 7, 19 -- } } } Received: from sentinel.ucr.edu (sentinel.ucr.edu } } } [138.23.226.228]) } } } 7, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } } } ESMTP id l4HGZqYO015701 } } } 7, 19 -- for {microscopy-at-microscopy.com} ; Thu, 17 May 2007 } } } 11:35:52 -0500 } } } 7, 19 -- Received: from [138.23.185.162] (igppb116g.ucr.edu } } } [138.23.185.162]) } } } 7, 19 -- by sentinel.ucr.edu (MOS 3.6.6-GR) } } } 7, 19 -- with ESMTP id ELY89098 (AUTH via LOGINBEFORESMTP) } } } 7, 19 -- for {microscopy-at-microscopy.com} ; } } } 7, 19 -- Thu, 17 May 2007 09:35:47 -0700 (PDT) } } } 7, 19 -- Mime-Version: 1.0 (Apple Message framework v752.2) 7, 19 -- } } } Content-Transfer-Encoding: 7bit 7, 19 -- Message-Id: } } } {8A1BE97E-989A-4B5C-A8FB-162874C89F6C-at-ucr.edu} } } } 7, 19 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; } } } format=flowed 7, 19 -- To: microscopy-at-microscopy.com 7, 19 -- From: } } } KN Bozhilov {bozhilov-at-ucr.edu} 7, 19 -- Subject: C2 apertures 7, 19 } } } -- Date: Thu, 17 May 2007 09:35:46 -0700 7, 19 -- X-Mailer: Apple } } } Mail (2.752.2) 7, 19 -- X-Junkmail-Whitelist: YES (by domain } } } whitelist at } } } sentinel.ucr.edu) } } } ==============================End of - } } } Headers============================== } } } } } } } } } ==============================Original } } } Headers============================== } } } 16, 38 -- From john.mardinly-at-intel.com Thu May 17 11:49:21 2007 16, } } } 38 -- Received: from mga09.intel.com (mga09.intel.com } } } [134.134.136.24]) } } } 16, 38 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } } } ESMTP id l4HGnKlY027440 } } } 16, 38 -- for {Microscopy-at-msa.microscopy.com} ; Thu, 17 May 2007 } } } 11:49:21 -0500 } } } 16, 38 -- Received: from fmsmga002.fm.intel.com ([10.253.24.26]) } } } 16, 38 -- by orsmga102.jf.intel.com with ESMTP; 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Thu, 17 15, 40 -- May 2007 } } 16:03:40 } } -0700 15, 40 -- Received: from scsmsx415.amr.corp.intel.com } } ([10.3.90.34]) by 15, 40 -- scsmsx411.amr.corp.intel.com with } } Microsoft SMTPSVC(6.0.3790.1830); Thu, 17 15, 40 -- May 2007 } } 16:03:40 } } -0700 15, 40 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 15, 40 } } -- Content-Class: urn:content-classes:message 15, 40 -- MIME- } } Version: } } 1.0 15, 40 -- Content-Type: text/plain; charset="us-ascii" } } 15, 40 -- Subject: RE: [Microscopy] RE: C2 apertures 15, 40 -- Date: } } Thu, 17 May 2007 16:03:39 -0700 15, 40 -- Message-ID: } } {F3CB8931ABF8294DB889E977150CAD687BC87B-at-scsmsx415.amr.corp.intel.com} } } 15, 40 -- In-Reply-To: {6A414FCC-B6A4-4273-AE52-2D5AE229B5D2-at-ucr.edu} } } 15, 40 -- X-MS-Has-Attach: } } 15, 40 -- X-MS-TNEF-Correlator: } } 15, 40 -- Thread-Topic: [Microscopy] RE: C2 apertures 15, 40 -- } } Thread-Index: AceYpf0XDwx27eZbSi6nsHj548GM0QAMMmuQ } } 15, 40 -- From: "Mardinly, John" {john.mardinly-at-intel.com} 15, 40 -- } } To: "KN Bozhilov" {bozhilov-at-ucr.edu} , {microscopy-at-microscopy.com} 15, } } 40 -- X-OriginalArrivalTime: 17 May 2007 23:03:40.0519 (UTC) } } FILETIME=[9BF3DF70:01C798D7] 15, 40 -- Content-Transfer-Encoding: } } 8bit } } 15, 40 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } } ns.microscopy.com id l4HN3f3W028250 ==============================End } } of - Headers============================== } } Hendrik O. Colijn colijn.1-at-osu.edu } OSU Campus Electron Optics Facility (614) 292-0674 } 040 Fontana Labs, 116 W. 19th Ave www.ceof.ohio-state.edu } } } ==============================Original } Headers============================== } 10, 26 -- From colijn.1-at-osu.edu Thu May 17 19:58:07 2007 10, 26 -- } Received: } from ER6S1.ENG.OHIO-STATE.EDU (er6s1.ecr6.ohio-state.edu } [164.107.76.2]) } 10, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id } l4I0w6Eh031712 } 10, 26 -- for {microscopy-at-microscopy.com} ; Thu, 17 May 2007 19:58:07 } -0500 } 10, 26 -- Received: from CONVERSION-DAEMON.er6s1.eng.ohio-state.edu } by 10, } 26 -- er6s1.eng.ohio-state.edu (PMDF V6.2-1x11 #31056) 10, 26 -- id } {01MGPFLZBYK0AA1TPQ-at-er6s1.eng.ohio-state.edu} for 10, 26 -- } microscopy-at-microscopy.com; Thu, 17 May 2007 20:58:06 -0400 (EDT) } 10, 26 -- } Received: from HOC3.osu.edu 10, 26 -- } (d149-67-33-174.col.wideopenwest.com } [67.149.174.33]) 10, 26 -- by er6s1.eng.ohio-state.edu (PMDF } V6.2-1x11 } #31056) 10, 26 -- with ESMTPA id } {01MGPFLY1E2MAAP4UD-at-er6s1.eng.ohio-state.edu} ; Thu, 10, 26 -- 17 } May 2007 } 20:58:05 -0400 (EDT) 10, 26 -- Date: Thu, 17 May 2007 20:57:59 } -0400 10, 26 } -- From: "Hendrik O. Colijn" {colijn.1-at-osu.edu} 10, 26 -- Subject: Re: } [Microscopy] C2 apertures 10, 26 -- In-reply-to: } {200705172305.l4HN5u9b030905-at-ns.microscopy.com} } 10, 26 -- Sender: colijn-at-er6s1.ecr6.ohio-state.edu 10, 26 -- To: } john.mardinly-at-intel.com 10, 26 -- Cc: Microscopy Listserver } {microscopy-at-microscopy.com} 10, 26 -- Message-id: } {7.0.1.0.2.20070517204654.0037cc80-at-osu.edu} } 10, 26 -- MIME-version: 1.0 } 10, 26 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 10, 26 -- } Content-type: text/plain; charset=us-ascii; format=flowed 10, 26 -- } X-Env-From: auth/colijn.1-at-osu.edu 10, 26 -- References: } {200705172305.l4HN5u9b030905-at-ns.microscopy.com} } ==============================End of - } Headers============================== } } } ==============================Original } Headers============================== } 19, 21 -- From walck-at-southbaytech.com Thu May 17 22:45:14 2007 } 19, 21 -- Received: from flpvm23.prodigy.net (flpvm23.prodigy.net } [207.115.20.53]) } 19, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id l4I3jD7V025675 } 19, 21 -- for {Microscopy-at-microscopy.com} ; Thu, 17 May 2007 } 22:45:13 -0500 } 19, 21 -- X-ORBL: [64.169.217.123] } 19, 21 -- Received: from dynamicbl8uno3 } (adsl-64-169-217-123.dsl.lsan03.pacbell.net [64.169.217.123]) } 19, 21 -- by flpvm23.prodigy.net (8.13.8 out.dk.spool/8.13.8) with } ESMTP id l4I3jNEK032553 } 19, 21 -- for {Microscopy-at-microscopy.com} ; Thu, 17 May 2007 } 20:45:23 -0700 } 19, 21 -- From: "Scott Walck" {walck-at-southbaytech.com} } 19, 21 -- To: {Microscopy-at-microscopy.com} } 19, 21 -- Subject: RE: [Microscopy] Re: C2 apertures } 19, 21 -- Date: Thu, 17 May 2007 20:45:39 -0700 } 19, 21 -- Message-ID: {004501c798ff$010c1540$7801a8c0-at-dynamicbl8uno3} } 19, 21 -- MIME-Version: 1.0 } 19, 21 -- Content-Type: text/plain; } 19, 21 -- charset="US-ASCII" } 19, 21 -- Content-Transfer-Encoding: 7bit } 19, 21 -- X-Mailer: Microsoft Office Outlook 11 } 19, 21 -- In-Reply-To: {200705180100.l4I10jMW005753-at-ns.microscopy.com} } 19, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 } 19, 21 -- Thread-Index: AceY5/gPj1VTNd0cR1S3iDOLy0KoSgAFpArg } ==============================End of - } Headers==============================
==============================Original Headers============================== 8, 21 -- From bozhilov-at-ucr.edu Fri May 18 10:31:41 2007 8, 21 -- Received: from sentoku.ucr.edu (sentoku.ucr.edu [138.23.226.244]) 8, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4IFVauD020453 8, 21 -- for {microscopy-at-microscopy.com} ; Fri, 18 May 2007 10:31:39 -0500 8, 21 -- Received: from [138.23.185.162] (igppb116g.ucr.edu [138.23.185.162]) 8, 21 -- by sentoku.ucr.edu (MOS 3.6.6-GR) 8, 21 -- with ESMTP id BKI19837 (AUTH via LOGINBEFORESMTP); 8, 21 -- Fri, 18 May 2007 08:31:28 -0700 (PDT) 8, 21 -- In-Reply-To: {200705180347.l4I3ljZX029085-at-ns.microscopy.com} 8, 21 -- References: {200705180347.l4I3ljZX029085-at-ns.microscopy.com} 8, 21 -- Mime-Version: 1.0 (Apple Message framework v752.2) 8, 21 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 8, 21 -- Message-Id: {E7D75C44-AC1B-40AA-9052-B7D943393B59-at-ucr.edu} 8, 21 -- Cc: walck-at-southbaytech.com 8, 21 -- Content-Transfer-Encoding: 7bit 8, 21 -- From: KN Bozhilov {bozhilov-at-ucr.edu} 8, 21 -- Subject: Re: [Microscopy] C2 apertures 8, 21 -- Date: Fri, 18 May 2007 08:31:28 -0700 8, 21 -- To: microscopy-at-microscopy.com 8, 21 -- X-Mailer: Apple Mail (2.752.2) 8, 21 -- X-Junkmail-Whitelist: YES (by domain whitelist at sentoku.ucr.edu) ==============================End of - Headers==============================
In that case, PICT would indeed be the format of choice. I found some information on the web that said the PICT format CAN include JPEG compression. That may be a user defined option, then.
Do you know if the iMovie format that is used before extracting he inividual frames uses mPEG compression?
mike
Michael Bode, Ph.D.
General Manager
OLYMPUS SOFT IMAGING SOLUTIONS 12596 West Bayaud Ave #300 Lakewood, CO 80228 USA Tel.: +1 (303) 234-9270 Fax.: +1 (303) 234-9271 E-mail: Mike.Bode-at-olympus-sis.com www.olympus-sis.com
-----Original Message----- X-from: tracy [mailto:trazy-at-umn.edu] Sent: Friday, May 18, 2007 09:26 To: Mike Bode; microscopy-at-microscopy.com
OK...
Now I'm a little confused.........
Because I was taught that a Pic file, like a screen shot is only a rasterized 72 dpi image.
So I threw a 6400 dpi image into iMovie, and saved as a pic file, and then opened in photoshop, and it says it's a 640 by 480 .............72 dpi image
Was the conversion on the import into iMovie, or is this what I've always found with pic files, 72 dpi cause that's what the Mac does with a pic file?
So I save as JPEG, and it's a 243x144 pixel image and still only 72 dpi.
If I use the OS X grab-it function for a screen shot, it actually saves it in TIFF, but again, only 72 bit dpi.
In which case it wouldn't make any difference just to use the System application "Grab-it", select to capture selection, and save using Grab-it, as both would be 72 dpi, and there would be no more conversion involved
Lou Ann
trazy-at-umn.edu wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } } The PICT files captured through iMovie are lossless, so this would be } the flavor of choice. You can then easily convert PICT to TIFF if } you wish. } } Cheers, } } -- } Tracy E. Anderson } }
==============================Original Headers============================== 16, 19 -- From lamiller-at-uiuc.edu Fri May 18 10:45:23 2007 16, 19 -- Received: from expredir5.cites.uiuc.edu (expredir5.cites.uiuc.edu [128.174.5.96]) 16, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4IFjMCi007315 16, 19 -- for {microscopy-at-microscopy.com} ; Fri, 18 May 2007 10:45:23 -0500 16, 19 -- Received: from redkite.cvm.uiuc.edu (redkite.cvm.uiuc.edu [130.126.18.157]) 16, 19 -- by expredir5.cites.uiuc.edu (8.14.1/8.14.1) with ESMTP id l4IFjL03025767 16, 19 -- for {microscopy-at-microscopy.com} ; Fri, 18 May 2007 10:45:21 -0500 (CDT) 16, 19 -- Message-ID: {464DCA12.9010409-at-uiuc.edu} 16, 19 -- Date: Fri, 18 May 2007 10:45:22 -0500 16, 19 -- From: Lou Ann Miller {lamiller-at-uiuc.edu} 16, 19 -- Reply-To: lamiller-at-uiuc.edu 16, 19 -- User-Agent: Thunderbird 2.0.0.0 (Macintosh/20070326) 16, 19 -- MIME-Version: 1.0 16, 19 -- To: microscopy-at-microscopy.com 16, 19 -- Subject: Re: [Microscopy] Re: viaWWW: Need Help From Imaging Experts 16, 19 -- References: {200705181526.l4IFQg4i007630-at-ns.microscopy.com} 16, 19 -- In-Reply-To: {200705181526.l4IFQg4i007630-at-ns.microscopy.com} 16, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 16, 19 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
iMovie deals with NTSC (TV) resolution (unless you're working in HDTV), which is 640x480 at 72dpi. So yes, any large image file brought into iMovie will be scaled down accordingly.
Presumably, the original poster is working with a video captured with NTSC resolution, so there would be no loss when capturing individual frames.
Cheers,
- Tracy E. Anderson Microscopist and digital imaging specialist Imaging Center University of Minnesota College of Biological Sciences Phone: 612.624.3454 Fax: 612.624.1799 http://www.cbs.umn.edu/ic/ -- "Science and art belong to the whole world, and before them vanish the barriers of nationality." - Goethe --
On 18 May 2007, lamiller-at-uiuc.edu wrote: } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } OK... } } Now I'm a little confused......... } } Because I was taught that a Pic file, like a screen shot is only a } rasterized 72 dpi image. } } } So I threw a 6400 dpi image into iMovie, and saved as a pic file, and } then opened in photoshop, and it says it's a 640 by 480 .............72 } dpi image } } } Was the conversion on the import into iMovie, or is this what I've } always found with pic files, 72 dpi cause that's what the Mac does with } a pic file? } } So I save as JPEG, and it's a 243x144 pixel image and still only 72 dpi. } } If I use the OS X grab-it function for a screen shot, it actually saves } it in TIFF, but again, only 72 bit dpi. } } In which case it wouldn't make any difference just to use the System } application "Grab-it", select to capture selection, and save using } Grab-it, as both would be 72 dpi, and there would be no more conversion } involved } } } } Lou Ann } } } trazy-at-umn.edu wrote: } } } ---------------------------------------------------------------------------- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } ---------------------------------------------------------------------------- } } } } } } The PICT files captured through iMovie are lossless, so this would be } } the flavor of choice. You can then easily convert PICT to TIFF if } } you wish. } } } } Cheers, } } } } -- } } Tracy E. Anderson } } } } } } } ==============================Original Headers============================== } 16, 19 -- From lamiller-at-uiuc.edu Fri May 18 10:45:23 2007 } 16, 19 -- Received: from expredir5.cites.uiuc.edu (expredir5.cites.uiuc.edu } [128.174.5.96]) } 16, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } l4IFjMCi007315 } 16, 19 -- for {microscopy-at-microscopy.com} ; Fri, 18 May 2007 10:45:23 -0500 } 16, 19 -- Received: from redkite.cvm.uiuc.edu (redkite.cvm.uiuc.edu } [130.126.18.157]) } 16, 19 -- by expredir5.cites.uiuc.edu (8.14.1/8.14.1) with ESMTP id } l4IFjL03025767 } 16, 19 -- for {microscopy-at-microscopy.com} ; Fri, 18 May 2007 10:45:21 -0500 } (CDT) } 16, 19 -- Message-ID: {464DCA12.9010409-at-uiuc.edu} } 16, 19 -- Date: Fri, 18 May 2007 10:45:22 -0500 } 16, 19 -- From: Lou Ann Miller {lamiller-at-uiuc.edu} } 16, 19 -- Reply-To: lamiller-at-uiuc.edu } 16, 19 -- User-Agent: Thunderbird 2.0.0.0 (Macintosh/20070326) } 16, 19 -- MIME-Version: 1.0 } 16, 19 -- To: microscopy-at-microscopy.com } 16, 19 -- Subject: Re: [Microscopy] Re: viaWWW: Need Help From Imaging } Experts } 16, 19 -- References: {200705181526.l4IFQg4i007630-at-ns.microscopy.com} } 16, 19 -- In-Reply-To: {200705181526.l4IFQg4i007630-at-ns.microscopy.com} } 16, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed } 16, 19 -- Content-Transfer-Encoding: 7bit } ==============================End of - } Headers============================== }
==============================Original Headers============================== 8, 20 -- From trazy-at-umn.edu Fri May 18 11:04:23 2007 8, 20 -- Received: from mta-w2.tc.umn.edu (mta-w2.tc.umn.edu [134.84.119.6]) 8, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4IG4NAT028201 8, 20 -- for {microscopy-at-microscopy.com} ; Fri, 18 May 2007 11:04:23 -0500 8, 20 -- Received: from barricade.software.umn.edu (barricade.software.umn.edu [128.101.65.74]) 8, 20 -- by mta-w2.tc.umn.edu (UMN smtpd) with ESMTP 8, 20 -- for {microscopy-at-microscopy.com} ; Fri, 18 May 2007 11:04:23 -0500 (CDT) 8, 20 -- X-Umn-Remote-Mta: [N] barricade.software.umn.edu [128.101.65.74] #+LO+TR 8, 20 -- Received: (from nobody-at-localhost) 8, 20 -- by barricade.software.umn.edu (8.12.11/8.12.8) id l4IG4Mna030948; 8, 20 -- Fri, 18 May 2007 11:04:22 -0500 8, 20 -- Message-Id: {200705181604.l4IG4Mna030948-at-barricade.software.umn.edu} 8, 20 -- Date: Fri, 18 May 2007 11:04:22 CDT 8, 20 -- From: tracy {trazy-at-umn.edu} 8, 20 -- Subject: Re: [Microscopy] viaWWW: Need Help From Imaging Experts 8, 20 -- To: microscopy-at-microscopy.com 8, 20 -- MIME-Version: 1.0 8, 20 -- Content-Type: TEXT/plain; CHARSET=US-ASCII 8, 20 -- X-Tick-Nemesis: American Maid 8, 20 -- X-remote-user-ip: 160.94.80.149 ==============================End of - Headers==============================
In that case, Grab-it can save directly to TIFF but would it be the same or worse than pict as a screen shot?
Lou Ann
trazy-at-umn.edu wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } iMovie deals with NTSC (TV) resolution (unless you're working in HDTV), which } is 640x480 at 72dpi. So yes, any large image file brought into iMovie will } be scaled down accordingly. } } Presumably, the original poster is working with a video captured with NTSC } resolution, so there would be no loss when capturing individual frames. } } Cheers, } } - } Tracy E. Anderson } Microscopist and digital imaging specialist } Imaging Center } University of Minnesota } College of Biological Sciences } Phone: 612.624.3454 } Fax: 612.624.1799 } http://www.cbs.umn.edu/ic/ } -- } "Science and art belong to the whole world, and before them vanish the } barriers of nationality." - Goethe } -- } } } On 18 May 2007, lamiller-at-uiuc.edu wrote: } } } } } } } } ---------------------------------------------------------------------------- } } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } } } ---------------------------------------------------------------------------- } } } OK... } } } } Now I'm a little confused......... } } } } Because I was taught that a Pic file, like a screen shot is only a } } rasterized 72 dpi image. } } } } } } So I threw a 6400 dpi image into iMovie, and saved as a pic file, and } } then opened in photoshop, and it says it's a 640 by 480 .............72 } } dpi image } } } } } } Was the conversion on the import into iMovie, or is this what I've } } always found with pic files, 72 dpi cause that's what the Mac does with } } a pic file? } } } } So I save as JPEG, and it's a 243x144 pixel image and still only 72 dpi. } } } } If I use the OS X grab-it function for a screen shot, it actually saves } } it in TIFF, but again, only 72 bit dpi. } } } } In which case it wouldn't make any difference just to use the System } } application "Grab-it", select to capture selection, and save using } } Grab-it, as both would be 72 dpi, and there would be no more conversion } } involved } } } } } } } } Lou Ann } } } } } } trazy-at-umn.edu wrote: } } } ---------------------------------------------------------------------------- } } } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } } } } America } } } } To Subscribe/Unsubscribe -- } } } } http://www.microscopy.com/MicroscopyListserver } } } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } } ---------------------------------------------------------------------------- } } } } The PICT files captured through iMovie are lossless, so this would be } } } the flavor of choice. You can then easily convert PICT to TIFF if } } } you wish. } } } } } } Cheers, } } } } } } -- } } } Tracy E. Anderson } } } } } } } } } } } } } ==============================Original } } } Headers============================== } } } 16, 19 -- From lamiller-at-uiuc.edu Fri May 18 10:45:23 2007 } } 16, 19 -- Received: from expredir5.cites.uiuc.edu (expredir5.cites.uiuc.edu } } [128.174.5.96]) } } 16, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } } l4IFjMCi007315 } } 16, 19 -- for {microscopy-at-microscopy.com} ; Fri, 18 May 2007 10:45:23 -0500 } } 16, 19 -- Received: from redkite.cvm.uiuc.edu (redkite.cvm.uiuc.edu } } [130.126.18.157]) } } 16, 19 -- by expredir5.cites.uiuc.edu (8.14.1/8.14.1) with ESMTP id } } l4IFjL03025767 } } 16, 19 -- for {microscopy-at-microscopy.com} ; Fri, 18 May 2007 10:45:21 -0500 } } (CDT) } } 16, 19 -- Message-ID: {464DCA12.9010409-at-uiuc.edu} } } 16, 19 -- Date: Fri, 18 May 2007 10:45:22 -0500 } } 16, 19 -- From: Lou Ann Miller {lamiller-at-uiuc.edu} } } 16, 19 -- Reply-To: lamiller-at-uiuc.edu } } 16, 19 -- User-Agent: Thunderbird 2.0.0.0 (Macintosh/20070326) } } 16, 19 -- MIME-Version: 1.0 } } 16, 19 -- To: microscopy-at-microscopy.com } } 16, 19 -- Subject: Re: [Microscopy] Re: viaWWW: Need Help From Imaging } } Experts } } 16, 19 -- References: {200705181526.l4IFQg4i007630-at-ns.microscopy.com} } } 16, 19 -- In-Reply-To: {200705181526.l4IFQg4i007630-at-ns.microscopy.com} } } 16, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed } } 16, 19 -- Content-Transfer-Encoding: 7bit } } ==============================End of - } } Headers============================== } } } } } } } } ==============================Original Headers============================== } 8, 20 -- From trazy-at-umn.edu Fri May 18 11:04:23 2007 } 8, 20 -- Received: from mta-w2.tc.umn.edu (mta-w2.tc.umn.edu [134.84.119.6]) } 8, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4IG4NAT028201 } 8, 20 -- for {microscopy-at-microscopy.com} ; Fri, 18 May 2007 11:04:23 -0500 } 8, 20 -- Received: from barricade.software.umn.edu (barricade.software.umn.edu [128.101.65.74]) } 8, 20 -- by mta-w2.tc.umn.edu (UMN smtpd) with ESMTP } 8, 20 -- for {microscopy-at-microscopy.com} ; Fri, 18 May 2007 11:04:23 -0500 (CDT) } 8, 20 -- X-Umn-Remote-Mta: [N] barricade.software.umn.edu [128.101.65.74] #+LO+TR } 8, 20 -- Received: (from nobody-at-localhost) } 8, 20 -- by barricade.software.umn.edu (8.12.11/8.12.8) id l4IG4Mna030948; } 8, 20 -- Fri, 18 May 2007 11:04:22 -0500 } 8, 20 -- Message-Id: {200705181604.l4IG4Mna030948-at-barricade.software.umn.edu} } 8, 20 -- Date: Fri, 18 May 2007 11:04:22 CDT } 8, 20 -- From: tracy {trazy-at-umn.edu} } 8, 20 -- Subject: Re: [Microscopy] viaWWW: Need Help From Imaging Experts } 8, 20 -- To: microscopy-at-microscopy.com } 8, 20 -- MIME-Version: 1.0 } 8, 20 -- Content-Type: TEXT/plain; CHARSET=US-ASCII } 8, 20 -- X-Tick-Nemesis: American Maid } 8, 20 -- X-remote-user-ip: 160.94.80.149 } ==============================End of - Headers============================== } }
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Lou Ann Miller, MT(ASCP) Service Supervisor Center for Microscopic Imaging College of Veterinary Medicine Rm 1204 VMBSB 2001 S Lincoln Ave Urbana, IL 61802
217/244-1567 http://treefrog.cvm.uiuc.edu
==============================Original Headers============================== 10, 19 -- From lamiller-at-uiuc.edu Fri May 18 11:18:19 2007 10, 19 -- Received: from expredir6.cites.uiuc.edu (expredir6.cites.uiuc.edu [128.174.5.97]) 10, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4IGIJ6L007437 10, 19 -- for {microscopy-at-microscopy.com} ; Fri, 18 May 2007 11:18:19 -0500 10, 19 -- Received: from redkite.cvm.uiuc.edu (redkite.cvm.uiuc.edu [130.126.18.157]) 10, 19 -- by expredir6.cites.uiuc.edu (8.14.1/8.14.1) with ESMTP id l4IGIJZ1010676 10, 19 -- for {microscopy-at-microscopy.com} ; Fri, 18 May 2007 11:18:19 -0500 (CDT) 10, 19 -- Message-ID: {464DD1CB.60309-at-uiuc.edu} 10, 19 -- Date: Fri, 18 May 2007 11:18:19 -0500 10, 19 -- From: Lou Ann Miller {lamiller-at-uiuc.edu} 10, 19 -- Reply-To: lamiller-at-uiuc.edu 10, 19 -- User-Agent: Thunderbird 2.0.0.0 (Macintosh/20070326) 10, 19 -- MIME-Version: 1.0 10, 19 -- To: microscopy-at-microscopy.com 10, 19 -- Subject: Re: [Microscopy] Re: viaWWW: Need Help From Imaging Experts 10, 19 -- References: {200705181605.l4IG50Io029204-at-ns.microscopy.com} 10, 19 -- In-Reply-To: {200705181605.l4IG50Io029204-at-ns.microscopy.com} 10, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 10, 19 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Download Graphic Converter from http://www.lemkesoft.com It will convert PICT files to TIFF or any of a plethora of other file formats. $30 shareware, free 30 day trial, nearly as powerful as Photoshop and *much* easier to use.
Phil
} I see what he means , just tried iMovie on OS X 10.3. } } } It seems to me though, that about 5 years ago, it DID let me save to } TIFF and there were more options, perhaps trying to get an Old copy of } iMovie? or try on an old machine if you still have OS 9? } } Also can you open the clip in Quicktime Pro? Maybe Quicktime Pro will } let you pull a tiff off of it, it too will save a frame. } } Not a great solution, but if you are in a pinch..... } } } } Lou Ann } } Mike.Bode-at-olympus-sis.com wrote: } } } In that case I would suggest to ask Apple. Contact them and tell them } } that for reasons of scientific Ethics you need the images in TIFF format } } (or any other format that you think might work), and ask them if they } } have a converter for their iMovie Product. They claim to be super } } user-friendly. Put them to the test. } } } } mike } } } } Michael Bode, Ph.D. } } } } General Manager } } } } OLYMPUS SOFT IMAGING SOLUTIONS } } 12596 West Bayaud Ave #300 } } Lakewood, CO 80228 } } USA } } Tel.: +1 (303) 234-9270 } } Fax.: +1 (303) 234-9271 } } E-mail: Mike.Bode-at-olympus-sis.com } } www.olympus-sis.com } } } } -----Original Message----- } } X-from: Sharon Bledsoe [mailto:sbledsoe-at-iupui.edu] } } Sent: Friday, May 18, 2007 06:12 } } To: Mike Bode } } Subject: [Microscopy] RE: viaWWW: Need Help From Imaging Experts } } } } IF I had a choice I would choose to save my images as TIFF's, however, } } the iMovie program will not let me. iMoive will only let me save as } } PICT or JPEG. } } } } And I am not tied to the Mac, I'm chained to it. } } } Hi Sharon, } } } } } } This may be a bit more fundamental than what you are looking for, but } } } here goes: } } } } } } If you go to the MSA website, you can find a statement about ethical } } } image processing that in essence says that images need to be stored } } } uncompressed. JPEG is a format that includes lossy compression, and I } } } believe that PICT can also contain lossy compression in the form of } } } JPEG. Both therefore don't seem to be the best choices. } } } } } } I also found this on the web: } } } } } } In the next version of its operating system, MacOS X, Apple has decided } } } } } } } } } } to replace PICT by PDF. This means that the use of PICT for exchanging } } } data will slowly diminish. } } } } } } It looks like sooner or later you will have to find a different choice } } } anyway. I would look at the whole chain of acquisition to see if you } } } cannot switch to something else. } } } } } } Why can't you use anything else? Does it have to do with the original } } } file format that can only be read by iMovie? Or are you tied to the Mac } } } } } } } } } } platform? } } } } } } mike } } } } } } } } } Michael Bode, Ph.D. } } } } } } General Manager } } } } } } OLYMPUS SOFT IMAGING SOLUTIONS } } } 12596 West Bayaud Ave #300 } } } Lakewood, CO 80228 } } } USA } } } Tel.: +1 (303) 234-9270 } } } Fax.: +1 (303) 234-9271 } } } E-mail: Mike.Bode-at-olympus-sis.com } } } www.olympus-sis.com } } } } Email: sbledsoe-at-iupui.edu } } } Name: Sharon Bledsoe } } } } } } Title-Subject: [Filtered] Need Help From Imaging Experts } } } } } } Question: Here is my problem: I can only use Mac iMovie to edit my } } } digital video. I can not do this on a PC. ONLY iMovie. To save a } } } single frame from the video, my choices are either PICT or JPEG format. } } } } } } } } } My Question is: To save an origianl image, which is the best format, } } } PICT or JPEG? } } } } } } I'm not interested in hearing about Tiffs, BMPs or any other format as } } } that is not one of my choices.
} ~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Lou Ann Miller, MT(ASCP) } Service Supervisor } Center for Microscopic Imaging } College of Veterinary Medicine } Rm 1204 VMBSB } 2001 S Lincoln Ave } Urbana, IL 61802 } } 217/244-1567 } http://treefrog.cvm.uiuc.edu
-- Philip Oshel Microscopy Facility Supervisor Biology Department 024C Brooks Hall Central Michigan University Mt. Pleasant, MI 48859 voice: (989) 774-3576 dept. fax: (989) 774-3462
==============================Original Headers============================== 5, 22 -- From oshel1pe-at-cmich.edu Fri May 18 11:18:31 2007 5, 22 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 5, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4IGIV8s007806 5, 22 -- for {Microscopy-at-microscopy.com} ; Fri, 18 May 2007 11:18:31 -0500 5, 22 -- Received: from egatea.central.cmich.local ([141.209.15.74]) 5, 22 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id l4IGfgqo015838 5, 22 -- for {Microscopy-at-microscopy.com} ; Fri, 18 May 2007 12:41:42 -0400 5, 22 -- Received: from [141.209.160.249] ([141.209.160.249]) by egatea.central.cmich.local with Microsoft SMTPSVC(6.0.3790.1830); 5, 22 -- Fri, 18 May 2007 12:18:26 -0400 5, 22 -- Mime-Version: 1.0 5, 22 -- Message-Id: {f06230907c2738198fb3e-at-[141.209.160.249]} 5, 22 -- In-Reply-To: {200705181522.l4IFMenS031360-at-ns.microscopy.com} 5, 22 -- References: {200705181522.l4IFMenS031360-at-ns.microscopy.com} 5, 22 -- Date: Fri, 18 May 2007 12:18:23 -0400 5, 22 -- To: Microscopy-at-microscopy.com 5, 22 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 5, 22 -- Subject: [Microscopy] Re: viaWWW: Need Help From Imaging Experts 5, 22 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 5, 22 -- X-OriginalArrivalTime: 18 May 2007 16:18:26.0671 (UTC) FILETIME=[2A2ED7F0:01C79968] 5, 22 -- X-CanItPRO-Stream: default 5, 22 -- X-Spam-Score: -4 () L_EXCH_MF 5, 22 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
The key to understanding the mechanism is that the structure of Ruthenium Tetroxide is the same as that of Osmium Tetroxide, a central heavy metal bound by double bonds to 4 oxygens. The reaction is the same as for Osmium when it interacts with organic material - breaking of one of the double bonds by the target and then cross linking of the target with the RuO4. Contrast is the result of added density of the heavy metal to the target.
Hayat is relatively quiet about the chemical reaction in his Fixation for Electron Microscopy. I seem to have my copy of his Positive Staining for Electron Microscopy at home so I don't know if he even covers it as a stain, my memory is that he doesn't. It is also not covered in Lewis and Knight's Staining Methods for Sectioned Material.
Paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-9354 Cell:204-781-1502 Fax:204-789-3926
==============================Original Headers============================== 12, 20 -- From paul_hazelton-at-umanitoba.ca Fri May 18 11:47:35 2007 12, 20 -- Received: from electra.cc.umanitoba.ca (electra.cc.umanitoba.ca [130.179.16.23]) 12, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4IGlXxl009903 12, 20 -- for {microscopy-at-microscopy.com} ; Fri, 18 May 2007 11:47:34 -0500 12, 20 -- Received: from umanitoba.ca (net40.medmb.umanitoba.ca [140.193.11.160]) 12, 20 -- (authenticated bits=0) 12, 20 -- by electra.cc.umanitoba.ca (8.13.0/8.13.0) with ESMTP id l4IGlR08019062; 12, 20 -- Fri, 18 May 2007 11:47:28 -0500 (CDT) 12, 20 -- Message-ID: {464DD81F.7090906-at-umanitoba.ca} 12, 20 -- Date: Fri, 18 May 2007 11:45:19 -0500 12, 20 -- From: paul r hazelton {paul_hazelton-at-umanitoba.ca} 12, 20 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.4) Gecko/20030624 Netscape/7.1 (ax) 12, 20 -- X-Accept-Language: en-us, en 12, 20 -- MIME-Version: 1.0 12, 20 -- To: pforcen-at-unizar.es, Microscopy Listserver {microscopy-at-microscopy.com} 12, 20 -- Subject: Re: [Microscopy] viaWWW: TEM and RuO4 12, 20 -- References: {200705181329.l4IDTJwU014370-at-ns.microscopy.com} 12, 20 -- In-Reply-To: {200705181329.l4IDTJwU014370-at-ns.microscopy.com} 12, 20 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 12, 20 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
As this discussion proceeds, please remember the pixel count is what's important and not the size of the image in inches or the DPI.
If you make that image bigger on the screen it will give you a few more pixels in the grabbed image, but where did those pixels come from? You have not changed anything in the native resolution of the movie images. If the image was originally 640x480, new pixels will not magically multiple.
In fact, if you don't increase the pixel count an integral number of times (e.g., 1280x960) you may introduce questions due to mapping. A new pixel will likely consist of contributions of multiple old pixels, or if you prefer, a single old pixel will contribute to multiple new pixels. Teh details depend on how the software magnifies small images. Still, you are not creating new information, so leave the image size at 640x480.
Personally, I still have little use for the DPI (actually PPI) value. Sure it will interact with some document programs to insert an image at the "proper size". I still prefer just knowing the pixel count and the image width and I can do the math myself. DPI just seems to confuse the matter for most folks.
Warren
-----Original Message----- X-from: lamiller-at-uiuc.edu [mailto:lamiller-at-uiuc.edu] Sent: Friday, May 18, 2007 11:26 AM To: wesaia-at-iastate.edu
I guess I should qualify the question...... I can make the image bigger on my screen while in iMovie.
Would then taking a bigger image as TIFF than one can with the save frame command, with Grab-it be of any benefit?
Such as making it smaller and increasing the dpi in photoshop mathmatically?
==============================Original Headers============================== 13, 32 -- From wesaia-at-iastate.edu Fri May 18 12:26:22 2007 13, 32 -- Received: from mailhub-3.iastate.edu (mailhub-3.iastate.edu [129.186.140.13]) 13, 32 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4IHQLnB022290 13, 32 -- for {Microscopy-at-msa.microscopy.com} ; Fri, 18 May 2007 12:26:22 -0500 13, 32 -- Received: from mailout-2.iastate.edu (mailout-2.iastate.edu [129.186.140.2]) 13, 32 -- by mailhub-3.iastate.edu (8.12.11.20060614/8.12.10) with SMTP id l4IHQLNq015119; 13, 32 -- Fri, 18 May 2007 12:26:21 -0500 13, 32 -- Received: from (owa.eng.iastate.edu [129.186.23.85]) by mailout-2.iastate.edu with smtp 13, 32 -- id 4485_885b9c9c_0564_11dc_97fe_001372578af6; 13, 32 -- Fri, 18 May 2007 12:23:45 -0500 13, 32 -- Received: from maire.eng.iastate.edu ([10.10.196.69]) by owa.eng.iastate.edu with Microsoft SMTPSVC(6.0.3790.3959); 13, 32 -- Fri, 18 May 2007 12:26:21 -0500 13, 32 -- Content-class: urn:content-classes:message 13, 32 -- MIME-Version: 1.0 13, 32 -- Content-Type: text/plain; 13, 32 -- charset="US-ASCII" 13, 32 -- Subject: RE: [Microscopy] viaWWW: Need Help From Imaging Experts 13, 32 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 13, 32 -- Date: Fri, 18 May 2007 12:27:13 -0500 13, 32 -- Message-ID: {16A330AC32056A40B32842EC4BB8D72701A175D4-at-maire.eng.iastate.edu} 13, 32 -- In-Reply-To: {200705181625.l4IGPZcP027497-at-ns.microscopy.com} 13, 32 -- X-MS-Has-Attach: 13, 32 -- X-MS-TNEF-Correlator: 13, 32 -- Thread-Topic: [Microscopy] viaWWW: Need Help From Imaging Experts 13, 32 -- Thread-Index: AceZaStFbxoPucVZRPGHKUo0hWluwQABeJzQ 13, 32 -- References: {200705181625.l4IGPZcP027497-at-ns.microscopy.com} 13, 32 -- From: "Straszheim, Warren E [CCE E]" {wesaia-at-iastate.edu} 13, 32 -- To: "MSA" {Microscopy-at-msa.microscopy.com} 13, 32 -- Cc: {lamiller-at-uiuc.edu} 13, 32 -- X-OriginalArrivalTime: 18 May 2007 17:26:21.0672 (UTC) FILETIME=[A712A680:01C79971] 13, 32 -- Content-Transfer-Encoding: 8bit 13, 32 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4IHQLnB022290 ==============================End of - Headers==============================
As Paul, I'm working from memory. However, I recall (hopefully accurate) information on mechanisms from Sawyer and Grubbs or Trent. I believe that the mechanisms for OsO4 and RuO4 oxidation of polyolefins is different. OsO4 adds across C=C bonds and does not react with C-C bonds at all. RuO4, on the other hand, scissions C-C or C=C bonds to produce oxygenated chain ends.
Regards,
Disclaimer: The comments and opinions are those of the author alone and do not reflect any official position by his employer.
Gary M. Brown Microscopy Specialist ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: 281 834 2387 fax: 281 834 2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
"Happiness equals reality minus expectations." Tom Magliozzi
paul_hazelton-at- umanitoba.ca To gary.m.brown-at-exxonmobil.com 05/18/07 11:50 cc AM Subject [Microscopy] Re: viaWWW: TEM and Please respond RuO4 to paul_hazelton-at- umanitoba.ca
The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
Patricia
The key to understanding the mechanism is that the structure of Ruthenium Tetroxide is the same as that of Osmium Tetroxide, a central heavy metal bound by double bonds to 4 oxygens. The reaction is the same as for Osmium when it interacts with organic material - breaking of one of the double bonds by the target and then cross linking of the target with the RuO4. Contrast is the result of added density of the heavy metal to the target.
Hayat is relatively quiet about the chemical reaction in his Fixation for Electron Microscopy. I seem to have my copy of his Positive Staining for Electron Microscopy at home so I don't know if he even covers it as a stain, my memory is that he doesn't. It is also not covered in Lewis and Knight's Staining Methods for Sectioned Material.
Paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-9354 Cell:204-781-1502 Fax:204-789-3926
==============================Original Headers============================== 12, 20 -- From paul_hazelton-at-umanitoba.ca Fri May 18 11:47:35 2007 12, 20 -- Received: from electra.cc.umanitoba.ca (electra.cc.umanitoba.ca [130.179.16.23]) 12, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4IGlXxl009903 12, 20 -- for {microscopy-at-microscopy.com} ; Fri, 18 May 2007 11:47:34 -0500 12, 20 -- Received: from umanitoba.ca (net40.medmb.umanitoba.ca [140.193.11.160]) 12, 20 -- (authenticated bits=0) 12, 20 -- by electra.cc.umanitoba.ca (8.13.0/8.13.0) with ESMTP id l4IGlR08019062; 12, 20 -- Fri, 18 May 2007 11:47:28 -0500 (CDT) 12, 20 -- Message-ID: {464DD81F.7090906-at-umanitoba.ca} 12, 20 -- Date: Fri, 18 May 2007 11:45:19 -0500 12, 20 -- From: paul r hazelton {paul_hazelton-at-umanitoba.ca} 12, 20 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.4) Gecko/20030624 Netscape/7.1 (ax) 12, 20 -- X-Accept-Language: en-us, en 12, 20 -- MIME-Version: 1.0 12, 20 -- To: pforcen-at-unizar.es, Microscopy Listserver {microscopy-at-microscopy.com} 12, 20 -- Subject: Re: [Microscopy] viaWWW: TEM and RuO4 12, 20 -- References: {200705181329.l4IDTJwU014370-at-ns.microscopy.com} 12, 20 -- In-Reply-To: {200705181329.l4IDTJwU014370-at-ns.microscopy.com} 12, 20 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 12, 20 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
==============================Original Headers============================== 34, 20 -- From gary.m.brown-at-exxonmobil.com Fri May 18 13:00:48 2007 34, 20 -- Received: from hoespc01.exxonmobil.com (hoespc01.exxonmobil.com [158.35.223.1]) 34, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4II0mqd002284 34, 20 -- for {microscopy-at-microscopy.com} ; Fri, 18 May 2007 13:00:48 -0500 34, 20 -- Received: from dalnmg04.na.xom.com (dalnmg04.na.xom.com [131.126.97.123]) 34, 20 -- by hoespc01.exxonmobil.com (Switch-3.1.11/Switch-3.1.11) with ESMTP id l4II0lK9027174; 34, 20 -- Fri, 18 May 2007 13:00:47 -0500 (CDT) 34, 20 -- In-Reply-To: {200705181650.l4IGoNfM013243-at-ns.microscopy.com} 34, 20 -- Subject: Re: [Microscopy] Re: viaWWW: TEM and RuO4 34, 20 -- Importance: 34, 20 -- To: microscopy-at-microscopy.com 34, 20 -- Cc: paul_hazelton-at-umanitoba.ca, pforcen-at-unizar.es 34, 20 -- X-Mailer: Lotus Notes 652HF109 January 14, 2005 34, 20 -- Message-ID: {OF6708A824.46B88797-ON862572DF.0061EB53-862572DF.0062F223-at-exxonmobil.com} 34, 20 -- From: gary.m.brown-at-exxonmobil.com 34, 20 -- Date: Fri, 18 May 2007 13:00:43 -0500 34, 20 -- X-MIMETrack: Serialize by Router on Dalnmg04.na.xom.com/S/ExxonMobil(652FP1HF193|March 34, 20 -- 02, 2006) at 05/18/2007 01:00:47 PM 34, 20 -- MIME-Version: 1.0 34, 20 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
Yes, that is the good question "What does iMovie do with the image when it is spread on the screen?"
An automatic .... it just spreads the pixels, actually may not be accurate.
I have a small box when I marginalize iMovie on my computer screen, say 4-5 inches ( also I do have a $4K Sony high resolution Pro-video camera), yet, as I do a lot of movies to DVD using iMovie, on a large TV screen, the product of iMovie does not appear to be pulling apart a 72 dpi small box., that's going from a 4 inch box to a 23 inch screen with only a slight loss of resolution, and that doesn't seem to fit the pulling apart of what we think of as a normal, email, web page weight images we know as 72 dpi.
So does iMovie just spit out the 72 dpi on the frame picture export work, and give back better stuff with more resolution with it's movie processing when the source was higher resolution? Or is the eye being fooled by the ~33 frames per second presentation?
I'll have to play with that when I get home,
Lou Ann
Straszheim, Warren E [CCE E] wrote: } As this discussion proceeds, please remember the pixel count is what's } important and not the size of the image in inches or the DPI. } } If you make that image bigger on the screen it will give you a few more } pixels in the grabbed image, but where did those pixels come from? You } have not changed anything in the native resolution of the movie images. } If the image was originally 640x480, new pixels will not magically } multiple. } } In fact, if you don't increase the pixel count an integral number of } times (e.g., 1280x960) you may introduce questions due to mapping. A new } pixel will likely consist of contributions of multiple old pixels, or if } you prefer, a single old pixel will contribute to multiple new pixels. } Teh details depend on how the software magnifies small images. Still, } you are not creating new information, so leave the image size at } 640x480. } } Personally, I still have little use for the DPI (actually PPI) value. } Sure it will interact with some document programs to insert an image at } the "proper size". I still prefer just knowing the pixel count and the } image width and I can do the math myself. DPI just seems to confuse the } matter for most folks. } } Warren } }
==============================Original Headers============================== 20, 19 -- From lamiller-at-uiuc.edu Fri May 18 13:46:48 2007 20, 19 -- Received: from expredir6.cites.uiuc.edu (expredir6.cites.uiuc.edu [128.174.5.97]) 20, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4IIkmAj014594 20, 19 -- for {microscopy-at-microscopy.com} ; Fri, 18 May 2007 13:46:48 -0500 20, 19 -- Received: from redkite.cvm.uiuc.edu (redkite.cvm.uiuc.edu [130.126.18.157]) 20, 19 -- by expredir6.cites.uiuc.edu (8.14.1/8.14.1) with ESMTP id l4IIkmjJ015748 20, 19 -- for {microscopy-at-microscopy.com} ; Fri, 18 May 2007 13:46:48 -0500 (CDT) 20, 19 -- Message-ID: {464DF498.3070506-at-uiuc.edu} 20, 19 -- Date: Fri, 18 May 2007 13:46:48 -0500 20, 19 -- From: Lou Ann Miller {lamiller-at-uiuc.edu} 20, 19 -- Reply-To: lamiller-at-uiuc.edu 20, 19 -- User-Agent: Thunderbird 2.0.0.0 (Macintosh/20070326) 20, 19 -- MIME-Version: 1.0 20, 19 -- To: microscopy-at-microscopy.com 20, 19 -- Subject: Re: [Microscopy] viaWWW: Need Help From Imaging Experts 20, 19 -- References: {200705181625.l4IGPZcP027497-at-ns.microscopy.com} {16A330AC32056A40B32842EC4BB8D72701A175D4-at-maire.eng.iastate.edu} 20, 19 -- In-Reply-To: {16A330AC32056A40B32842EC4BB8D72701A175D4-at-maire.eng.iastate.edu} 20, 19 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 20, 19 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
I would appreciate any insight that the microscopy community might have regarding safe practices and tools for cutting and trimming polymer samples. Our lab members generally use single- and double-edge razor blades for small jobs. Any suggestions on these or other tools, blade guards, etc. will be appreciated.
Thanks,
Disclaimer: The comments and opinions are those of the author alone and do not reflect any official position by ExxonMobil Chemical Company.
Gary M. Brown Microscopy Specialist ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: 281 834 2387 fax: 281 834 2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
"Happiness equals reality minus expectations." Tom Magliozzi
==============================Original Headers============================== 8, 18 -- From gary.m.brown-at-exxonmobil.com Fri May 18 15:33:00 2007 8, 18 -- Received: from dalspc01.exxonmobil.com (dalspc01.exxonmobil.com [131.126.223.1]) 8, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4IKX0E7029976 8, 18 -- for {microscopy-at-microscopy.com} ; Fri, 18 May 2007 15:33:00 -0500 8, 18 -- Received: from dalnmg04.na.xom.com (dalnmg04.na.xom.com [131.126.97.123]) 8, 18 -- by dalspc01.exxonmobil.com (Switch-3.1.11/Switch-3.1.11) with ESMTP id l4IKWwGS025609 8, 18 -- for {microscopy-at-microscopy.com} ; Fri, 18 May 2007 15:33:00 -0500 (CDT) 8, 18 -- Subject: Cutting tools and hazards 8, 18 -- Importance: 8, 18 -- To: microscopy-at-microscopy.com 8, 18 -- X-Mailer: Lotus Notes 652HF109 January 14, 2005 8, 18 -- Message-ID: {OF81EDDCD5.9E4BDC38-ON862572DF.00702DBB-862572DF.0070E207-at-exxonmobil.com} 8, 18 -- From: gary.m.brown-at-exxonmobil.com 8, 18 -- Date: Fri, 18 May 2007 15:32:56 -0500 8, 18 -- X-MIMETrack: Serialize by Router on Dalnmg04.na.xom.com/S/ExxonMobil(652FP1HF193|March 8, 18 -- 02, 2006) at 05/18/2007 03:33:00 PM 8, 18 -- MIME-Version: 1.0 8, 18 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
Gary's right, the reaction of the C=O with unsaturated C=C double bonds - such as seen with olefins results in a five atom circular structure between the original 2 carbons, and two O's and the Os from the OsO4. Also, as he points out, with olefins, as more OsO4 gets into the act you can see scissions of the C-C bond. At least that is the theory, I'm just a scope jockey/structural/molecular virologist. In my humble opinion it takes a very different brain than mine to understand that organic chemistry stuff.
Paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-9354 Cell:204-781-1502 Fax:204-789-3926
==============================Original Headers============================== 8, 21 -- From paul_hazelton-at-umanitoba.ca Fri May 18 15:33:42 2007 8, 21 -- Received: from electra.cc.umanitoba.ca (electra.cc.umanitoba.ca [130.179.16.23]) 8, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4IKXgvh030903 8, 21 -- for {microscopy-at-microscopy.com} ; Fri, 18 May 2007 15:33:42 -0500 8, 21 -- Received: from umanitoba.ca (net40.medmb.umanitoba.ca [140.193.11.160]) 8, 21 -- (authenticated bits=0) 8, 21 -- by electra.cc.umanitoba.ca (8.13.0/8.13.0) with ESMTP id l4IKXfvt021963; 8, 21 -- Fri, 18 May 2007 15:33:41 -0500 (CDT) 8, 21 -- Message-ID: {464E0D25.3040502-at-umanitoba.ca} 8, 21 -- Date: Fri, 18 May 2007 15:31:33 -0500 8, 21 -- From: paul r hazelton {paul_hazelton-at-umanitoba.ca} 8, 21 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.4) Gecko/20030624 Netscape/7.1 (ax) 8, 21 -- X-Accept-Language: en-us, en 8, 21 -- MIME-Version: 1.0 8, 21 -- To: pforcen-at-unizar.es 8, 21 -- CC: Microscopy Listserver {microscopy-at-microscopy.com} 8, 21 -- Subject: Re: [Microscopy] viaWWW: TEM and RuO4 8, 21 -- References: {200705181329.l4IDTJwU014370-at-ns.microscopy.com} 8, 21 -- In-Reply-To: {200705181329.l4IDTJwU014370-at-ns.microscopy.com} 8, 21 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 8, 21 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Apparently at one time, FEI sold sort of a super-TopHat condenser aperture that was essentially a number of condenser apertures stacked on each other with some sort of low-Z material in between. Perhaps that is the aperture you are describing. The amazingly high cost could be due to the obvious difficulty of aligning the stacked apertures.
Unless you need incredibly clean EDS spectra, try a normal Pt TopHat aperture or even a regular 250micron thick Pt condenser aperture. Of course, you could always try stacking some apertures yourself!
Cheers, Henk
At 07:05 PM 5/17/2007, you wrote:
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Hendrik O. Colijn colijn.1-at-osu.edu OSU Campus Electron Optics Facility (614) 292-0674 040 Fontana Labs, 116 W. 19th Ave www.ceof.ohio-state.edu
==============================Original Headers============================== 10, 25 -- From colijn.1-at-osu.edu Fri May 18 20:04:36 2007 10, 25 -- Received: from ER6S1.ENG.OHIO-STATE.EDU (er6s1.ecr6.ohio-state.edu [164.107.76.2]) 10, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4J14apL026604 10, 25 -- for {microscopy-at-microscopy.com} ; Fri, 18 May 2007 20:04:36 -0500 10, 25 -- Received: from CONVERSION-DAEMON.er6s1.eng.ohio-state.edu by 10, 25 -- er6s1.eng.ohio-state.edu (PMDF V6.2-1x11 #31056) 10, 25 -- id {01MGQU5CD2CGAA2DK4-at-er6s1.eng.ohio-state.edu} for 10, 25 -- microscopy-at-microscopy.com; Fri, 18 May 2007 21:04:35 -0400 (EDT) 10, 25 -- Received: from HOC3.osu.edu 10, 25 -- (d149-67-33-174.col.wideopenwest.com [67.149.174.33]) 10, 25 -- by er6s1.eng.ohio-state.edu (PMDF V6.2-1x11 #31056) 10, 25 -- with ESMTPA id {01MGQU5B3MCWA9MPCL-at-er6s1.eng.ohio-state.edu} ; Fri, 10, 25 -- 18 May 2007 21:04:34 -0400 (EDT) 10, 25 -- Date: Fri, 18 May 2007 20:50:33 -0400 10, 25 -- From: "Hendrik O. Colijn" {colijn.1-at-osu.edu} 10, 25 -- Subject: Re: [Microscopy] C2 apertures 10, 25 -- In-reply-to: {200705172305.l4HN5u9b030905-at-ns.microscopy.com} 10, 25 -- Sender: colijn-at-er6s1.ecr6.ohio-state.edu 10, 25 -- To: Microscopy Listserver {microscopy-at-microscopy.com} , bozhilov-at-ucr.edu 10, 25 -- Message-id: {7.0.1.0.2.20070518203334.04d3fb58-at-osu.edu} 10, 25 -- MIME-version: 1.0 10, 25 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 10, 25 -- Content-type: text/plain; charset=us-ascii; format=flowed 10, 25 -- X-Env-From: auth/colijn.1-at-osu.edu 10, 25 -- References: {200705172305.l4HN5u9b030905-at-ns.microscopy.com} ==============================End of - Headers==============================
Please, forget about the "dpi" rating of the image. What is important (as has been pointed out in a previous reply) is simply the _total number_ of pixels. After all, you can increase the dpi very simply by displaying the image on a tiny screen. The same number of pixels on fewer inches, so higher dpi. Of course, that did not improve the actual image content at all.
In a previous life, I wrote consumer level non-linear video editing software.
Usually, video editing software works in such a way that the final resolution is determined by the format you're exporting the movie to. If you're targetting a low-bandwidth "web movie", it is rendered to 320x240 or thereabouts. All source clips and images are re-rendered in this target resolution. If you added a high-resolution still image (from an 8 megapixel photo camera) in the middle of your movie, this image will be downsampled to the final image resolution, and not somehow stored in the movie in its original resolution.
If you target a more high-quality format such as DVD or even HD, all source clips and images are rendered into this higher resolution format. If your originals were in low quality, the resulting images are simply "stretched to fit". This is the same effect as when you enlarge your media player to full screen: there is no "extra information" - (after all, the media player would somehow have to "make it up" since it is not present in the movie data); the existing information is simply interpolated to give it a "smooth appearance". Added to the way human perception works for moving images, this gives an excellent _perceived_ quality.
But there's more.
If you're capturing movies with a digital camera, chances are very high that there is already some compression being performed during the capture. If it's a DV camera, the quality of these images is quite good; but if it's a modern MPEG-2 camera, you can expect lots of compression artefacts. Movies taken this way are simply not meant to be dissected frame-by-frame, as human perception for moving images and for still images is very different. This is extra visible if frames are captured interlaced (i.e. one field will contain all the odd lines, the next field all the even ones). This is great for moving images as this will actually increase the perceived quality, but looking at one such a field in a still will show hideous jagged edges.
So again, if you extract a single frame from a digitized, compressed movie, exporting it to uncompressed TIFF will not magically increase its quality. It will only not make it _worse_.
You will probably get better suggestions from the list if you don't limit our choices to "A or B", but tell us exactly what you're trying to accomplish.
Yes, that is the good question "What does iMovie do with the image when it is spread on the screen?"
An automatic .... it just spreads the pixels, actually may not be accurate.
I have a small box when I marginalize iMovie on my computer screen, say 4-5 inches ( also I do have a $4K Sony high resolution Pro-video camera), yet, as I do a lot of movies to DVD using iMovie, on a large TV screen, the product of iMovie does not appear to be pulling apart a 72 dpi small box., that's going from a 4 inch box to a 23 inch screen with only a slight loss of resolution, and that doesn't seem to fit the pulling apart of what we think of as a normal, email, web page weight images we know as 72 dpi.
So does iMovie just spit out the 72 dpi on the frame picture export work, and give back better stuff with more resolution with it's movie processing when the source was higher resolution? Or is the eye being fooled by the ~33 frames per second presentation?
I'll have to play with that when I get home,
Lou Ann
==============================Original Headers============================== 22, 27 -- From Sander.Stoks-at-fei.com Sat May 19 05:29:00 2007 22, 27 -- Received: from smtp-nl1.feico.com (smtp-nl1.feico.com [195.75.179.210]) 22, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4JASxSb014904 22, 27 -- for {Microscopy-at-Microscopy.Com} ; Sat, 19 May 2007 05:28:59 -0500 22, 27 -- X-WSS-ID: 0JIAAI5-02-1T7-01 22, 27 -- Received: from acht850.w2k.feico.com (unknown [10.150.55.50]) 22, 27 -- by smtp-nl1.feico.com (Tumbleweed MailGate) with SMTP id 8249D10F7FB5 22, 27 -- for {Microscopy-at-Microscopy.Com} ; Sat, 19 May 2007 03:30:05 -0700 (PDT) 22, 27 -- Received: From acht887.w2k.feico.com ([10.150.55.87]) by acht850.w2k.feico.com (WebShield SMTP v4.5 MR2); 22, 27 -- id 1179570538178; Sat, 19 May 2007 12:28:58 +0200 22, 27 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 22, 27 -- Content-class: urn:content-classes:message 22, 27 -- MIME-Version: 1.0 22, 27 -- Content-Type: text/plain; 22, 27 -- charset="iso-8859-1" 22, 27 -- Subject: RE: [Microscopy] Re: viaWWW: Need Help From Imaging Experts 22, 27 -- Date: Sat, 19 May 2007 12:28:57 +0200 22, 27 -- Message-ID: {9211E4E38C8130428654AD6A1081A2CD0BF4F064-at-acht887.w2k.feico.com} 22, 27 -- X-MS-Has-Attach: 22, 27 -- X-MS-TNEF-Correlator: 22, 27 -- Thread-Topic: [Microscopy] Re: viaWWW: Need Help From Imaging Experts 22, 27 -- Thread-Index: AceZg9TrsGFoaMvwSf6rL80oGpWu4gAeClts 22, 27 -- References: {200705181852.l4IIqwJe024402-at-ns.microscopy.com} 22, 27 -- From: "Stoks, Sander" {Sander.Stoks-at-fei.com} 22, 27 -- To: {Microscopy-at-Microscopy.Com} 22, 27 -- Content-Transfer-Encoding: 8bit 22, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4JASxSb014904 ==============================End of - Headers==============================
There is a thread currently discussing digital imaging and movies etc...
Let me take a moment to remind you (or reinforce with others have said or implied) namely : if you save a file in any "lossy" compressed format your changing as well as loosing data. This must be reported if you use any of this processed data for any scientific analysis.
The last time I checked there are some 60+ different image file formats.
Lossless formats include (but are not limited to ) the following: RAW, TIFF, BMP, PICT, PNG, PCX, EXR, SVG, TGA, JPEG2000
Lossy formats include: GIF, JPEG, MPEG, MOV, H263, Video
On top of this there are also lossy and lossless COMPRESSION methods, which may be encoded within/upon some of these formats. So life can get very complicated , you might use a lossless method and then compress the data using a lossy alogrithm.
The Microscopy Society of America (MSA) has defined a policy on ethical digital imaging, which is available on line under Reference/Education section of their WWW site (http://www.microscopy.org) . This ethical position applies to all digital images, be they: still , time series or movies.
While making movies from time series events etc.. is a valuable mechanism of looking through your data, (and I also do this frequently). Remember that digital imaging ethics requires that you store the original data using a procedure similiar to that documented below. If you analyze your data you should refer back to the original uncompressed data, never to the compressed data.
You also need to report these operation particuliarly if you downsample, compress etc your data. Consumer grade software does this routinely and likely does not tell you so. So be extremely careful when using some of this software for analysis your scientific data!
If you are recording directly to digital video, I would recommend that you record in RAW format store that data and then after the fact convert your images to some alternative display.
----------------------------------------------------------------------------------- Here is the MSA statement verbatum. -----------------------------------------------------------------------------------
The MSA position on digital image processing has been approved as follows:
"Ethical digital imaging requires that the original uncompressed image file be stored on archival media (e.g., CD-R) without any image manipulation or processing operation. All parameters of the production and acquisition of this file, as well as any subsequent processing steps, must be documented and reported to ensure reproducibility.
Generally, acceptable (non-reportable) imaging operations include gamma correction, histogram stretching, and brightness and contrast adjustments. All other operations (such as Unsharp-Masking, Gaussian Blur, etc.) must be directly identified by the author as part of the experimental methodology. However, for diffraction data or any other image data that is used for subsequent quantification, all imaging operations must be reported."
This policy was formulated by the Digital Image Processing & Ethics Group of the MSA Education Committee and was adopted as MSA policy at the Summer Council meeting August 2-3, 2003.
Nestor Your Friendly Neighborhood SysOp
==============================Original Headers============================== 20, 13 -- From zaluzec-at-microscopy.com Sat May 19 10:16:30 2007 20, 13 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) 20, 13 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4JFGTRx031039 20, 13 -- for {microscopy-at-microscopy.com} ; Sat, 19 May 2007 10:16:29 -0500 20, 13 -- Mime-Version: 1.0 20, 13 -- Message-Id: {p06240801c274b9743c66-at-[206.69.208.22]} 20, 13 -- In-Reply-To: {200705181545.l4IFjOOa007362-at-ns.microscopy.com} 20, 13 -- References: {200705181545.l4IFjOOa007362-at-ns.microscopy.com} 20, 13 -- Date: Sat, 19 May 2007 10:16:28 -0500 20, 13 -- To: microscopy-at-microscopy.com 20, 13 -- From: "Nestor J. Zaluzec" {zaluzec-at-microscopy.com} 20, 13 -- Subject: Digital Imaging, Digital Movies and Compression 20, 13 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
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Email: lamiller-at-uiuc.edu Name: Lou Ann Miller
Organization: U of Illinois
Title-Subject: [Filtered] Imaging
Question: Thanks Sander and Warren,
Those answers helped explain what happens with the video, which was what my question was if not stated very clearly, you don't have to beg me to give up on dpi, ;-)
But I was curious as to what exactly the video software was actually doing to the image when, and it's good to know what mixed resolution inputs and expected outputs will do, and that the camera is actually doing some compression ( like scanners might ) to the image.
I'm a videographer on weekends, basically putting our church services to DVD for shut ins ( we do pay a license for copyright) and the sermon movies online (stjohnsermons.org). What one does with what is dependent upon what works best for the task, and not the dpi. I don't use grab-it, but do use the export on the software.
For example, I capture action shots and dim lit areas far off (balcony is a long LONG ways back) images off the video frames than I do with my very very nice 8 mega pixel camera even with the zoom lens, better meaning I got the data I wanted with out blurring...... confirmation, school graduations etc etc. Also, since even my digital camera is a reflex, and sounds like the paparazzi in a place with excellent sound carrying capability and tiled floor place, video suits better than even the 8 mega pixel very nice lens system in some situations. For other uses, the camera wins over the video frames hands down.
I'm sure in microscopy it is the same, what imaging technique is used, very much dependent on what works best for the best results.
So usage and results, and what is being done to the image, do win over the dpi. I know people who buy pocket cameras the size of 1/2 a wallet with huge mega pixel value, but their images don't turn out as well as they expect for buying that mega pixel value ...... because they don't use a good lens system...and probably other reasons you can tell me about for sure :-)
Thanks for the discussion! I've enjoyed it.
Lou Ann
(email failed from my home computer so submitting via web)
Note that BMP _used_ to be a lossless format, but since (I think) Windows 2000 a valid BMP file _can_ in fact contain a JPEG compressed (i.e., lossy) image. I've yet to run into one of those "in the wild".
Also: GIF does not use lossy compression (but is never used for serious image processing).
Regarding "RAW video": Suppose you are capturing full NTSC frames (there are different opinions on what the exact resolution is, but let's take 720x540 for the example) at 30 full frames per second (actually 29.97), in a raw format with three bytes per pixel this would take more than 32MB per second of footage. This would fill up a normal-sized hard disk quite rapidly. What's worse, up to a few years ago consumer grade hard disks couldn't even store data at this speed. Compression wasn't added to movie formats just to save space and costs; it was mainly because there simply wasn't any other way of doing real-time image acquisition otherwise. I don't know of any current (affordable) video cameras which can store their captured data in a raw format, but perhaps the state of the art has advanced more than I know.
Also, there's the problem that even if you pick a well-known image format and stay away from the "obscure" ones, you'll still run into spotty support even on major software platforms. For example, I regulartly get "complaints" from people claiming my TIFF exporter is broken because Word can't read the resulting TIFFs. Usually, that's because they've chosen 16-bit export, and Word only partially implements the (huge) TIFF specification and simply can't read all valid TIFFs. Likewise, Windows Explorer won't show any thumbnails for those images, because it only handles 8 bit-per-channel TIFFs.
The whole of "image postprocessing" is a bit of a gray area. If you use postprocessing to downsample an image you should report it, but if you select "binning 2" on your camera when doing the acquisition, you're in effect doing the same thing. Even worse, some consumer-grade cameras perform a host of image post processing _in the camera itself_ to make their images look better. It's not uncommon to see an "artificial sharpening filter" being performed to get away with a cheaper lens system.
I agree with reporting on the entire post-processing setup though, and with storing your original images in a lossless format, preferably TIFF, with sufficient bit depth (if you have a 10 bit TEM camera, don't store your images as BMP even if that format is listed as "lossless"). Even if some of your software can't correctly handle these original files, there are plenty of (free) conversion libraries around (many open source, so they will _stay_ around) which can convert it to any format you currently need.
Regards, Sander Stoks (not speaking for FEI)
________________________________
X-from: zaluzec-at-microscopy.com [mailto:zaluzec-at-microscopy.com] Sent: Sat 5/19/2007 5:27 PM To: Stoks, Sander
Colleagues
There is a thread currently discussing digital imaging and movies etc...
Let me take a moment to remind you (or reinforce with others have said or implied) namely : if you save a file in any "lossy" compressed format your changing as well as loosing data. This must be reported if you use any of this processed data for any scientific analysis.
The last time I checked there are some 60+ different image file formats.
Lossless formats include (but are not limited to ) the following: RAW, TIFF, BMP, PICT, PNG, PCX, EXR, SVG, TGA, JPEG2000
Lossy formats include: GIF, JPEG, MPEG, MOV, H263, Video
On top of this there are also lossy and lossless COMPRESSION methods, which may be encoded within/upon some of these formats. So life can get very complicated , you might use a lossless method and then compress the data using a lossy alogrithm.
The Microscopy Society of America (MSA) has defined a policy on ethical digital imaging, which is available on line under Reference/Education section of their WWW site (http://www.microscopy.org) . This ethical position applies to all digital images, be they: still , time series or movies.
While making movies from time series events etc.. is a valuable mechanism of looking through your data, (and I also do this frequently). Remember that digital imaging ethics requires that you store the original data using a procedure similiar to that documented below. If you analyze your data you should refer back to the original uncompressed data, never to the compressed data.
You also need to report these operation particuliarly if you downsample, compress etc your data. Consumer grade software does this routinely and likely does not tell you so. So be extremely careful when using some of this software for analysis your scientific data!
If you are recording directly to digital video, I would recommend that you record in RAW format store that data and then after the fact convert your images to some alternative display.
----------------------------------------------------------------------------------- Here is the MSA statement verbatum. -----------------------------------------------------------------------------------
The MSA position on digital image processing has been approved as follows:
"Ethical digital imaging requires that the original uncompressed image file be stored on archival media (e.g., CD-R) without any image manipulation or processing operation. All parameters of the production and acquisition of this file, as well as any subsequent processing steps, must be documented and reported to ensure reproducibility.
Generally, acceptable (non-reportable) imaging operations include gamma correction, histogram stretching, and brightness and contrast adjustments. All other operations (such as Unsharp-Masking, Gaussian Blur, etc.) must be directly identified by the author as part of the experimental methodology. However, for diffraction data or any other image data that is used for subsequent quantification, all imaging operations must be reported."
This policy was formulated by the Digital Image Processing & Ethics Group of the MSA Education Committee and was adopted as MSA policy at the Summer Council meeting August 2-3, 2003.
Nestor Your Friendly Neighborhood SysOp
==============================Original Headers============================== 20, 13 -- From zaluzec-at-microscopy.com Sat May 19 10:16:30 2007 20, 13 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) 20, 13 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4JFGTRx031039 20, 13 -- for {microscopy-at-microscopy.com} ; Sat, 19 May 2007 10:16:29 -0500 20, 13 -- Mime-Version: 1.0 20, 13 -- Message-Id: {p06240801c274b9743c66-at-[206.69.208.22]} 20, 13 -- In-Reply-To: {200705181545.l4IFjOOa007362-at-ns.microscopy.com} 20, 13 -- References: {200705181545.l4IFjOOa007362-at-ns.microscopy.com} 20, 13 -- Date: Sat, 19 May 2007 10:16:28 -0500 20, 13 -- To: microscopy-at-microscopy.com 20, 13 -- From: "Nestor J. Zaluzec" {zaluzec-at-microscopy.com} 20, 13 -- Subject: Digital Imaging, Digital Movies and Compression 20, 13 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
==============================Original Headers============================== 33, 27 -- From Sander.Stoks-at-fei.com Sat May 19 14:27:54 2007 33, 27 -- Received: from smtp-nl1.feico.com (smtp-nl1.feico.com [195.75.179.210]) 33, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4JJRrWE027424 33, 27 -- for {microscopy-at-microscopy.com} ; Sat, 19 May 2007 14:27:54 -0500 33, 27 -- X-WSS-ID: 0JIAZGF-02-6EK-01 33, 27 -- Received: from acht850.w2k.feico.com (unknown [10.150.55.50]) 33, 27 -- by smtp-nl1.feico.com (Tumbleweed MailGate) with SMTP id 9380110F96C8 33, 27 -- for {microscopy-at-microscopy.com} ; Sat, 19 May 2007 12:29:02 -0700 (PDT) 33, 27 -- Received: From acht887.w2k.feico.com ([10.150.55.87]) by acht850.w2k.feico.com (WebShield SMTP v4.5 MR2); 33, 27 -- id 1179602871426; Sat, 19 May 2007 21:27:51 +0200 33, 27 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 33, 27 -- Content-class: urn:content-classes:message 33, 27 -- MIME-Version: 1.0 33, 27 -- Content-Type: text/plain; 33, 27 -- charset="iso-8859-1" 33, 27 -- Subject: RE: [Microscopy] Digital Imaging, Digital Movies and Compression 33, 27 -- Date: Sat, 19 May 2007 21:27:51 +0200 33, 27 -- Message-ID: {9211E4E38C8130428654AD6A1081A2CD0BF4F065-at-acht887.w2k.feico.com} 33, 27 -- X-MS-Has-Attach: 33, 27 -- X-MS-TNEF-Correlator: 33, 27 -- Thread-Topic: [Microscopy] Digital Imaging, Digital Movies and Compression 33, 27 -- Thread-Index: AceaKjw331Fna805QeWBCv3yauCb1gAHUfuX 33, 27 -- References: {200705191527.l4JFRVaA018445-at-ns.microscopy.com} 33, 27 -- From: "Stoks, Sander" {Sander.Stoks-at-fei.com} 33, 27 -- To: {microscopy-at-microscopy.com} 33, 27 -- Content-Transfer-Encoding: 8bit 33, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4JJRrWE027424 ==============================End of - Headers==============================
The problem here is that saving in iMovie has compressed the data to 640 by 480 (or as several have pointed out 10 or 15 variants of this) So you start out with really awful resolution. THEN the data is compressed by imovie to be able to handle the movie without needing terabyte drives. The compression of the movie is in MPEG 2 if it is DVD compliant. The codec can vary here if you are not trying to have a format that can be played on your TV. (At last count my computer has about 50 different codecs)
Before we start talking about frames, your data has thrown away anything bigger than 640 by 480 and compressed it alot.( This is an understatement.. ALOT!!!) The data that you grab will be 640 by 480 compressed. If you use a lossless format as many have suggested then you won't lose any more. Any method that you now use will never, never, never make the data better than 640 by 480.
All you are doing is interpolating the data up. This will not improve anything. If you are not careful the interpolation may include sharpening and/or smoothing. You would therefore need to document this. The reason that this all gets so complicated is that many of the suggestions are trying to "fix" the loss of resolution that occured when the data was aquired in imovie and this just can't be done.
The 72 dpi is meaningless for alot of reasons 1. The 72 dpi came from the screen resolution of computers in 1990. 2. The dpi is only really descriptive when you are talking about output. 3. Dpi is meaningless as the dot is defined so many different ways that you really have no idea what the dot is at any given time.
If you are restricted to movie format then best way to do it is to record on mini dv format. The sony cameras have an internal hardware compressor that is DVD compliant. It records data at 40 Megabytes per second. This is about 10 times the data rate that "direct digital" methods can achieve. The digital tape can record MUCH faster than a hard drive. After recording you can bring the data into thew computer over the IEEE 1394 (firewire) input. This is a frame accurate data transfer that does not change the data in any way. You do not lose one bit of data. The frames that you then can save need to be saved in a lossless manner. I would suggest you use Adobe Premier because it is a Mac program and you can save in a large variety of lossless formats.
Please understand that the chip that does the compression in the Sony cameras is one of those best kept secrets. Sony developed an unbelievably powerful chip so that they could own the mini dv format. They won but unfortuneately for them consumers are much more interested in BluRay and HD DVD
The documentation for publication needs to describe the acquisition chain as you are not recording anything that is uncompressed. The MSA guidelines that Nestor pointed you to would require that you report that hardware/software system so that others can reproduce your results
A HD tv camera would improve things greatly but I have not had time to run any tests to see exactly what the bandwidth of the system is. Since you would be putting 4 times as much data onto the tape you might have to use a chip that compresses it more than the Sony does for NTSC/PAL format.
John
John M. Mackenzie, Jr., PhD
Professor of Microbiology Coordinator, Center for Electron Microscopy North Carolina State University Phone (919) 515-2664 Fax (919) 515-8293
lamiller-at-uiuc.edu wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } I guess I should qualify the question...... I can make the image } bigger on my screen while in iMovie. } } } Would then taking a bigger image as TIFF than one can with the save } frame command, with Grab-it be of any benefit? } } Such as making it smaller and increasing the dpi in photoshop mathmatically? } } } } } ==============================Original Headers============================== } 7, 19 -- From lamiller-at-uiuc.edu Fri May 18 11:23:35 2007 } 7, 19 -- Received: from expredir4.cites.uiuc.edu (expredir4.cites.uiuc.edu [128.174.5.187]) } 7, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4IGNYTp022176 } 7, 19 -- for {microscopy-at-microscopy.com} ; Fri, 18 May 2007 11:23:34 -0500 } 7, 19 -- Received: from redkite.cvm.uiuc.edu (redkite.cvm.uiuc.edu [130.126.18.157]) } 7, 19 -- by expredir4.cites.uiuc.edu (8.14.1/8.14.1) with ESMTP id l4IGNXRb002933 } 7, 19 -- for {microscopy-at-microscopy.com} ; Fri, 18 May 2007 11:23:34 -0500 (CDT) } 7, 19 -- Message-ID: {464DD305.9000109-at-uiuc.edu} } 7, 19 -- Date: Fri, 18 May 2007 11:23:33 -0500 } 7, 19 -- From: Lou Ann Miller {lamiller-at-uiuc.edu} } 7, 19 -- Reply-To: lamiller-at-uiuc.edu } 7, 19 -- User-Agent: Thunderbird 2.0.0.0 (Macintosh/20070326) } 7, 19 -- MIME-Version: 1.0 } 7, 19 -- To: microscopy-at-microscopy.com } 7, 19 -- Subject: Re: [Microscopy] Re: viaWWW: Need Help From Imaging Experts } 7, 19 -- References: {200705181526.l4IFQg4i007630-at-ns.microscopy.com} {464DCA12.9010409-at-uiuc.edu} } 7, 19 -- In-Reply-To: {464DCA12.9010409-at-uiuc.edu} } 7, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed } 7, 19 -- Content-Transfer-Encoding: 7bit } ==============================End of - Headers============================== }
==============================Original Headers============================== 16, 20 -- From john_mackenzie-at-ncsu.edu Sun May 20 00:57:04 2007 16, 20 -- Received: from ms-smtp-05.southeast.rr.com (ms-smtp-05.southeast.rr.com [24.25.9.104]) 16, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4K5v3UN009602 16, 20 -- for {Microscopy-at-microscopy.com} ; Sun, 20 May 2007 00:57:04 -0500 16, 20 -- Received: from [127.0.0.1] (cpe-065-190-163-148.nc.res.rr.com [65.190.163.148]) 16, 20 -- by ms-smtp-05.southeast.rr.com (8.13.6/8.13.6) with ESMTP id l4K5v8iY013900; 16, 20 -- Sun, 20 May 2007 01:57:10 -0400 (EDT) 16, 20 -- Message-ID: {464FE32F.8010407-at-ncsu.edu} 16, 20 -- Date: Sun, 20 May 2007 01:57:03 -0400 16, 20 -- From: John Mackenzie {john_mackenzie-at-ncsu.edu} 16, 20 -- User-Agent: Thunderbird 1.5.0.10 (Windows/20070221) 16, 20 -- MIME-Version: 1.0 16, 20 -- To: lamiller-at-uiuc.edu 16, 20 -- CC: Microscopy-at-microscopy.com 16, 20 -- Subject: Re: [Microscopy] viaWWW: Need Help From Imaging Experts 16, 20 -- References: {200705181630.l4IGUKOe006345-at-ns.microscopy.com} 16, 20 -- In-Reply-To: {200705181630.l4IGUKOe006345-at-ns.microscopy.com} 16, 20 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 16, 20 -- Content-Transfer-Encoding: 7bit 16, 20 -- X-Virus-Scanned: Symantec AntiVirus Scan Engine ==============================End of - Headers==============================
Let's say you have a scale of 0-100 for your vacuum gauge. You need 95 in order to get a full emission reading on the SEM. The roughing pump works fine, brings it up to about 30, and the diffusion pump kicks in and rapidly brings it up to 90.
Here's the symptom:
The emission current never comes on. I can hear the high pitched sound of the high voltage, and occasionally the emission will peak at 20-30%, but then quickly die down again. There are two things that I can think of that could be wrong. First, there is a capacitor in the emission circuitry that is going a little haywire. Second, the vacuum system is fluctuating just above and then just below optimal levels.
Would low diffusion pump oil create a symptom like that where the vacuum fluctuates (Incidentally, the vacuum meter doesn't move, but I'm guessing that it isn't stable because of the fluctuating emission current). I thought that having a low amount of diffusion pump oil would just result in a lower vacuum, not necessarily a fluctuating one.
Thanks in advance,
Justin A. Kraft
==============================Original Headers============================== 6, 26 -- From kraftpiano-at-gmail.com Sun May 20 00:58:45 2007 6, 26 -- Received: from wa-out-1112.google.com (wa-out-1112.google.com [209.85.146.183]) 6, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4K5wjM6010992 6, 26 -- for {microscopy-at-microscopy.com} ; Sun, 20 May 2007 00:58:45 -0500 6, 26 -- Received: by wa-out-1112.google.com with SMTP id j5so55876wah 6, 26 -- for {microscopy-at-microscopy.com} ; Sat, 19 May 2007 22:58:54 -0700 (PDT) 6, 26 -- DKIM-Signature: a=rsa-sha1; c=relaxed/relaxed; 6, 26 -- d=gmail.com; s=beta; 6, 26 -- h=domainkey-signature:received:received:message-id:date:from:to:subject:mime-version:content-type:content-transfer-encoding:content-disposition; 6, 26 -- b=MEblRa87yPpgxhDh03egD8rxfH412qeiIPyOFHdh4EYWf/0cHefy6RRZ3PVfz/8Z+VlnrubAVpjPRPsjmPfgZgYqIBZKnufxObUupwXUiYdzC6I1GMq7vtx8Rbiqlz3YWQzFpsDqGmlP4q4tstOdAHrXZFOYBGvE+kwfn+i8OtY= 6, 26 -- DomainKey-Signature: a=rsa-sha1; c=nofws; 6, 26 -- d=gmail.com; s=beta; 6, 26 -- h=received:message-id:date:from:to:subject:mime-version:content-type:content-transfer-encoding:content-disposition; 6, 26 -- b=IlEzF6lyRKtIfOKckhSb59JXIxncjxt4MKHuD2wtAbHOjOHOHIzL7EX0hBwovCBMJeOXdSVp6vwkpmoHl9p2GqVpVsFeGjSuo4PlpFLXhj4AVyl1th2YT/YttI/41nF59cm3VtYbarD5rAgVeri3oC4Ap+JzmsTIympWrcx+sAQ= 6, 26 -- Received: by 10.114.210.2 with SMTP id i2mr1863371wag.1179640733372; 6, 26 -- Sat, 19 May 2007 22:58:53 -0700 (PDT) 6, 26 -- Received: by 10.114.78.15 with HTTP; Sat, 19 May 2007 22:58:52 -0700 (PDT) 6, 26 -- Message-ID: {25e2b0d20705192258p24954f1fhad190396029ff6dd-at-mail.gmail.com} 6, 26 -- Date: Sun, 20 May 2007 01:58:52 -0400 6, 26 -- From: "Justin Kraft" {kraftpiano-at-gmail.com} 6, 26 -- To: microscopy-at-microscopy.com 6, 26 -- Subject: SEM: Quick vacuum question. 6, 26 -- MIME-Version: 1.0 6, 26 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 6, 26 -- Content-Transfer-Encoding: 7bit 6, 26 -- Content-Disposition: inline ==============================End of - Headers==============================
1- diffusion pump oil may be too low; use at main diff pump 100cc, column diff pump 35cc oil filling. Warm up pumps, drain, check liquid (should be white/ yellow, not brownish). Use same oil as had been inside the pump. If you do not know if for ex. Santovac had been used, you have to clean the pumps thoroughly. Use NEW O-rings (special silicon? because of heat) to seal pumps again. Never use the old O-rings on the diff pump drains again!!!
2- You rebuild the complete column / pumping lines. There will be a lot of water inside all lines. This may take some days of continuos pumping to get rid of.
3-I suppose you checked the filament. Is it OK? Did you get a reading from the socket of the wehnelt? Is this OK? Should be some ohms... If not, clean the "feets" ofr the filament with very fine sandpaper. Sometimes long stored old filaments build up insulating layers...
4- Try some very low accelerating voltage, 2 KV or so. Are the fluctuations gone? If so, check for cleanliness of cathode / anode and keep the instrument pumping for some days...
----- Original Message ----- X-from: {kraftpiano-at-gmail.com} To: {stefan.diller-at-t-online.de} Sent: Sunday, May 20, 2007 8:08 AM
Hi
I am afraid on first sight your problem does not look like a "quick vacuum question".
Lets look at your comments regarding the vacuum and DP. With a level of fluid that is too small the DP will pump in "spurts" as the fluid boils off and then has to wait for condensation before it may re cycle. In most cases this will be adequate to hold a specific vacuum but it may take longer to reach a good operating pressure. The position of most manufacturers vacuum gauges means that they only give a rough clue as to what is going on in the gun so they will not help here. Removing the gun cable and replacing it with a gauge is the only way to really find out what is happening in the gun area - an engineer task in most laboratories.
Much more to the point, and in my view the problem, is the high voltage situation. If the vacuum is not good enough for the high voltage to operate it will switch off. If a component is breaking down in the high voltage circuitry it will switch off. To isolate the high voltage system from the vacuum may help in the investigation.
Switch off the high voltage and remove the cable from the tank. Wrap the cable end in aluminium foil and cover the tank connection socket. Switch on the high voltage watch the emission meter and listen. Same sounds? Same meter reactions? Nothing changes then it is a tank problem. Different symptoms then it is a high voltage cable or gun vacuum problem.
Can we find out more?
Steve Chapman Protrain for EM training & consultancy world wide www.emcourses.com Tel +44 1280 816512 Fax +44 1280 814007 Mobile +44 7711 606967
----- Original Message ----- X-from: {kraftpiano-at-gmail.com} To: {protrain-at-emcourses.com} Sent: Sunday, May 20, 2007 7:00 AM
Hi all,
Here when we produce a 'video' time-lapse on our optical microscope systems we are actually collecting a series of uncompressed TIF 'photographs' at 12-bit (4096 grey levels) or similar in a propriety format. These images are therefore discrete and if we want to 'extract' a quality still image we simply export/save one from the list, normally in .jpg hi-res compressed format for articles etc.. where the slight loss of image quality is unimportant given the image size in print, or as original uncompressed TIF format for image analysis if that program can't read the original file format. The cameras or detectors we use are typically things like an £10,000 Hammamatsu Orca ER B&W one. Technically at maximum resolutions of around 1280x1280 this is only just over a 1 megapixel camera, but of course the camera has excellent sensitivity at low light for fluorescence samples and has very low noise (I can see room for future improvement in resolution though). Our 'video' image sequences will thus be created from a series of microphotographs of live cells obtained using a £80k to £250k optical time-lapse wide-field or confocal optical microscope system. Images being captured at a maximum rate of one every few seconds unless we are prepared to take a hit in resolution (e.g. binning pixels) and/or noise for capturing rapid intra-cellular activity.
Thus to produce a 'proper' DVD type video, we export these TIF image sequences into a .mov (Apple) or .avi (PC) format. We are often only concerned that this exported video looks very similar to the original TIF sequence collected by say Kinetic Imaging AQM, OpenLabs, Leica LCS, Bio-Rad Lasersharp etc. For lab presentations we go for maximum quality during compression (or even lossless) as a movie of 100 Mb+ is no problem for a modern PC during presentation, and generally we just use the 'export to movie/video' option on the microscope/camera image capture software. As Microsoft Office on the PC now inconveniently doesn't support .mov video any more we mainly use £25 Quicktime Pro [v6] to convert the Apple .mov to PC .avi format afterwards (normally with the setting to minimum compression & maximum quality as our video sequences aren't that long, say up 1,000 images maximum and often far far less). On the odd occasions that we require a 100Mb+ video to be compressed to under 4Mb (say for web support of a published article) we then turn to our single £200 Adobe Premiere Pro (v1.5) software and play around with the export settings until we just hit 4Mb limit and image quality is adequate to illustrate the point (and we still have the original lossless image sequences should anyone wish to view them).
QuickTime Pro (and probably iMovie) can't compete with professional video software for this type of thing, but as it's so simple to use we stick to QuickTime Pro for routine video conversion of .mov to .avi (only I really know anything about using Adobe Premiere Pro here). For Apple users you would probably benefit with something like Final Cut Studio 2 as the professional video editing option, or indeed Adobe Premiere CS3 that runs on Apples (to get all features you will need boot-camp as well, but I doubt that's necessary for single frame export or further video compression). Note that you can run QuickTime .mov in PowerPoint 2003, but you have to insert it as a link to QuickTime viewer, and most users here prefer the simplicity of converting the video sequence to .avi which runs natively within PowerPoint. Plus you have to add the .jpg, .avi or .mov suffix on export from an Apple to get the PC to notice what type of file it is [e.g. for auto-play and preview].
So as we often want to routinely export our 'video' images into hi-res 12-bit uncompressed TIFF [from a microscope], we got a decent (and expensive) microscope camera and capture in lossless TIFF type format in the first place as a video time-lapse sequence (rather than use a cheap NTSC/PAL video camera). Generally these cameras were supplied by the microscope manufactures and/or the image capture/processing software company (e.g. OpenLabs, Zeiss, Leica), and we were happy with their recommendations (it’s a small percent of the system total cost and they often write the camera drivers).
I would add that rather like applying complex image processor filters to images, it's far easier just to play around with the export quality settings in something like £25 QuickTime Pro or better still £200 Adobe Premiere CS3 and see how far you have to go to get an acceptable image. This is easier than trying to think too much and too deeply about what would work best. The software normally offers a big guide hint with terms like 'maximum quality' and 'maximum compression' or 'minimum file size & minimum quality'. What is acceptable naturally varies widely depending on what the image is for, but normally for us it will look very very much like the original TIF image on casual inspection.
But if you have a compressed video image sequence to start with, or you have them archived as uncompressed TIFF sequences elsewhere in an OpenLabs propriety .Liff or Leica LCS .Lei file/folder, you have far less worries about what you do - in both cases you have the unadulterated original anyway should anyone raise questions and you are often just providing the small article photo or highly compressed on-line video. Thus the exported frames quality is just set to simply what is acceptable for the task in hand (in terms of image quality, file storage size and what you want to show). It's very easy to see when you have gone too far in terms of compression, bit depth etc - but make sure your VDU monitor and graphics card are set to a correct high bit depth, brightness/contrast and large pixel number screen 'size' that can show the exported image/video correctly. Plus 12-bit B&W images need to be exported converted to 256 grey levels (8-bit) or 16-bit for Photoshop to work with them correctly (off-line image analysis programs expect 12-bit and so will have no problems with the original).
In most cases we only really need to use our uncompressed 12-bit TIFF sequences for image analysis as it's pretty essential for image intensity measurements and it's known pixel area is easier to calibrate for things like manual count per unit area and object area (plus it's in that format anyway). For home use I would love to keep all my colour SLR photographs in .raw or TIF format, but in practice have to store them in hi-quality jpeg to prevent the hard drive filling up in a few weeks (don't edit and save .jpg images repeatedly though as you will lose detail with each re-compression file save - that’s another advantage of TIF/RAW and a patient disposition when working on them).
With 'proper' DVD/Camcorder video, I do find that working with a Sony MiniDV Camcorder video using cheap [Nero 7 type] authoring software to capture and write a DVD in a standard home PC always seems to stick in some compression and nasty pixelisation somewhere, even when I say please don't do that at all (compared to the original tape image quality on a nominally 625 line PAL TV). It's also worse captured to PC via its USB2 station compared to Firewire lead (as stated in the manual). However when I use the analogue PAL output via S-Video lead from the MiniDV camcorder straight into the analogue AV input of a dedicated TV DVD recorder the DVD image quality in XP/SP is excellent (as good as standard PAL TV on casual viewing). Seems strange given the two times analogue to digital conversion involved, so I assume it's just that the dedicated £300 TV DVD recorder is doing a far better job with writing DVDs than my old PC, as that’s all it does for the money (it can't play Doom 3 or run Photoshop though). John's just gone into this aspect in more detail (and I'm not NTSC friendly except for making sure I can play Region 1 DVD film releases on my TV). I can't use my works copy of Premiere Pro on this home PC with my Sony camcorder to test lossless or lower compression rates (as suggested by John) as it's got an AMD older style Athlon CPU chip (not compatible).
I know the above is the exact inverse of the question (and tends to ignore it), but hopefully it will be on interest to a few list-servers (particularly to those of us who find the majority of the electron microscope threads merely 'quite interesting').
Regards
Keith J Morris
PS Adobe Premiere is great value as Create Suite three (CS3) Production Premium, where you get:
Photoshop Adobe After Effects (graphics and visual effects) Adobe Premiere® Pro CS3 (video editing) Adobe Photoshop® CS3 Extended (photo-editing) Adobe Flash CS3 Professional (interactive web and instructional content), Adobe Illustrator CS3 (drawing/illustration) Adobe Soundbooth™ CS3 (film audio) Adobe Encore® CS3 (DVD authoring, but I tend to use Nero 7).
Normally available very cheaply as the complete CS3 package (so getting the less useful programs is just a free bonus). CS3 isn't released yet though.
Just a shame that Adobe, unlike Microsoft, have no concept of user friendly software - e.g. Adobe help always discusses a command in almost enough detail without ever saying where you can actually find it, and in Indesign (DT Publisher) they put the spell checker on the left under Edit, instead of on the right somewhere like Office and everyone else, so that it takes 5 minutes to find it first time (sure they aren't related to Apple?).
----------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Institute of Ophthalmology UCL, London EC1V 9EL
-----Original Message----- X-from: john_mackenzie-at-ncsu.edu [mailto:john_mackenzie-at-ncsu.edu] Sent: 20 May 2007 07:07 To: keith.morris-at-ucl.ac.uk
Folks
The problem here is that saving in iMovie has compressed the data to 640 by 480 (or as several have pointed out 10 or 15 variants of this) So you start out with really awful resolution. THEN the data is compressed by imovie to be able to handle the movie without needing terabyte drives. The compression of the movie is in MPEG 2 if it is DVD compliant. The codec can vary here if you are not trying to have a format that can be played on your TV. (At last count my computer has about 50 different codecs)
Before we start talking about frames, your data has thrown away anything bigger than 640 by 480 and compressed it alot.( This is an understatement.. ALOT!!!) The data that you grab will be 640 by 480 compressed. If you use a lossless format as many have suggested then you won't lose any more. Any method that you now use will never, never, never make the data better than 640 by 480.
All you are doing is interpolating the data up. This will not improve anything. If you are not careful the interpolation may include sharpening and/or smoothing. You would therefore need to document this. The reason that this all gets so complicated is that many of the suggestions are trying to "fix" the loss of resolution that occured when the data was aquired in imovie and this just can't be done.
The 72 dpi is meaningless for alot of reasons 1. The 72 dpi came from the screen resolution of computers in 1990. 2. The dpi is only really descriptive when you are talking about output. 3. Dpi is meaningless as the dot is defined so many different ways that you really have no idea what the dot is at any given time.
If you are restricted to movie format then best way to do it is to record on mini dv format. The sony cameras have an internal hardware compressor that is DVD compliant. It records data at 40 Megabytes per second. This is about 10 times the data rate that "direct digital" methods can achieve. The digital tape can record MUCH faster than a hard drive. After recording you can bring the data into thew computer over the IEEE 1394 (firewire) input. This is a frame accurate data transfer that does not change the data in any way. You do not lose one bit of data. The frames that you then can save need to be saved in a lossless manner. I would suggest you use Adobe Premier because it is a Mac program and you can save in a large variety of lossless formats.
Please understand that the chip that does the compression in the Sony cameras is one of those best kept secrets. Sony developed an unbelievably powerful chip so that they could own the mini dv format. They won but unfortuneately for them consumers are much more interested in BluRay and HD DVD
The documentation for publication needs to describe the acquisition chain as you are not recording anything that is uncompressed. The MSA guidelines that Nestor pointed you to would require that you report that hardware/software system so that others can reproduce your results
A HD tv camera would improve things greatly but I have not had time to run any tests to see exactly what the bandwidth of the system is. Since you would be putting 4 times as much data onto the tape you might have to use a chip that compresses it more than the Sony does for NTSC/PAL format.
John
John M. Mackenzie, Jr., PhD
Professor of Microbiology Coordinator, Center for Electron Microscopy North Carolina State University Phone (919) 515-2664 Fax (919) 515-8293
lamiller-at-uiuc.edu wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } I guess I should qualify the question...... I can make the image } bigger on my screen while in iMovie. } } } Would then taking a bigger image as TIFF than one can with the save } frame command, with Grab-it be of any benefit? } } Such as making it smaller and increasing the dpi in photoshop mathmatically? } } } } } ==============================Original Headers============================== } 7, 19 -- From lamiller-at-uiuc.edu Fri May 18 11:23:35 2007 } 7, 19 -- Received: from expredir4.cites.uiuc.edu (expredir4.cites.uiuc.edu [128.174.5.187]) } 7, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4IGNYTp022176 } 7, 19 -- for {microscopy-at-microscopy.com} ; Fri, 18 May 2007 11:23:34 -0500 } 7, 19 -- Received: from redkite.cvm.uiuc.edu (redkite.cvm.uiuc.edu [130.126.18.157]) } 7, 19 -- by expredir4.cites.uiuc.edu (8.14.1/8.14.1) with ESMTP id l4IGNXRb002933 } 7, 19 -- for {microscopy-at-microscopy.com} ; Fri, 18 May 2007 11:23:34 -0500 (CDT) } 7, 19 -- Message-ID: {464DD305.9000109-at-uiuc.edu} } 7, 19 -- Date: Fri, 18 May 2007 11:23:33 -0500 } 7, 19 -- From: Lou Ann Miller {lamiller-at-uiuc.edu} } 7, 19 -- Reply-To: lamiller-at-uiuc.edu } 7, 19 -- User-Agent: Thunderbird 2.0.0.0 (Macintosh/20070326) } 7, 19 -- MIME-Version: 1.0 } 7, 19 -- To: microscopy-at-microscopy.com } 7, 19 -- Subject: Re: [Microscopy] Re: viaWWW: Need Help From Imaging Experts } 7, 19 -- References: {200705181526.l4IFQg4i007630-at-ns.microscopy.com} {464DCA12.9010409-at-uiuc.edu} } 7, 19 -- In-Reply-To: {464DCA12.9010409-at-uiuc.edu} } 7, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed } 7, 19 -- Content-Transfer-Encoding: 7bit } ==============================End of - Headers============================== }
==============================Original Headers============================== 16, 20 -- From john_mackenzie-at-ncsu.edu Sun May 20 00:57:04 2007 16, 20 -- Received: from ms-smtp-05.southeast.rr.com (ms-smtp-05.southeast.rr.com [24.25.9.104]) 16, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4K5v3UN009602 16, 20 -- for {Microscopy-at-microscopy.com} ; Sun, 20 May 2007 00:57:04 -0500 16, 20 -- Received: from [127.0.0.1] (cpe-065-190-163-148.nc.res.rr.com [65.190.163.148]) 16, 20 -- by ms-smtp-05.southeast.rr.com (8.13.6/8.13.6) with ESMTP id l4K5v8iY013900; 16, 20 -- Sun, 20 May 2007 01:57:10 -0400 (EDT) 16, 20 -- Message-ID: {464FE32F.8010407-at-ncsu.edu} 16, 20 -- Date: Sun, 20 May 2007 01:57:03 -0400 16, 20 -- From: John Mackenzie {john_mackenzie-at-ncsu.edu} 16, 20 -- User-Agent: Thunderbird 1.5.0.10 (Windows/20070221) 16, 20 -- MIME-Version: 1.0 16, 20 -- To: lamiller-at-uiuc.edu 16, 20 -- CC: Microscopy-at-microscopy.com 16, 20 -- Subject: Re: [Microscopy] viaWWW: Need Help From Imaging Experts 16, 20 -- References: {200705181630.l4IGUKOe006345-at-ns.microscopy.com} 16, 20 -- In-Reply-To: {200705181630.l4IGUKOe006345-at-ns.microscopy.com} 16, 20 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 16, 20 -- Content-Transfer-Encoding: 7bit 16, 20 -- X-Virus-Scanned: Symantec AntiVirus Scan Engine ==============================End of - Headers==============================
==============================Original Headers============================== 53, 25 -- From keith.morris-at-ucl.ac.uk Sun May 20 08:16:34 2007 53, 25 -- Received: from smtp3.global.net.uk (smtp3.global.net.uk [80.189.92.91]) 53, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4KDGXWr016038 53, 25 -- for {Microscopy-at-microscopy.com} ; Sun, 20 May 2007 08:16:34 -0500 53, 25 -- Received: from [91.125.104.192] (helo=loungepc) 53, 25 -- by smtp3.global.net.uk with esmtp (Exim 4.66 (FreeBSD)) 53, 25 -- (envelope-from {keith.morris-at-ucl.ac.uk} ) 53, 25 -- id 1HplGr-0007gL-Fy 53, 25 -- for Microscopy-at-microscopy.com; Sun, 20 May 2007 14:16:25 +0100 53, 25 -- From: "Keith Morris" {keith.morris-at-ucl.ac.uk} 53, 25 -- To: {Microscopy-at-microscopy.com} 53, 25 -- References: {200705200606.l4K66rbX022528-at-ns.microscopy.com} 53, 25 -- Subject: WWW: Need Help From Imaging Experts 53, 25 -- Date: Sun, 20 May 2007 14:16:38 +0100 53, 25 -- Message-ID: {000901c79ae1$19b993e0$0401a8c0-at-loungepc} 53, 25 -- MIME-Version: 1.0 53, 25 -- Content-Type: text/plain; 53, 25 -- charset="iso-8859-1" 53, 25 -- X-Mailer: Microsoft Office Outlook 11 53, 25 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 53, 25 -- Thread-Index: AceapSEpYrJE2MCARbyu9aesTWQ4AAAB9jgg 53, 25 -- In-Reply-To: {200705200606.l4K66rbX022528-at-ns.microscopy.com} 53, 25 -- Authenticated-Sender: 53, 25 -- Content-Transfer-Encoding: 8bit 53, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4KDGXWr016038 ==============================End of - Headers==============================
I have gotten several off-line replies to my request for information on cutting tools and hazards. I appreciate any and all information and encourage replies to sent to the listserver for all to benefit. If, for privacy concerns, you choose to reply off-line, I will summarize your anonymous contribution with other feedback.
One last note: I will appreciate any comments, suggestions, concerns regarding any kind of cutting or preparation tool. This in an area where many of us have expertise that should be shared with the community at large.
Thanks,
Gary M. Brown Microscopy Specialist ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: 281 834 2387 fax: 281 834 2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
"Happiness equals reality minus expectations." Tom Magliozzi
==============================Original Headers============================== 8, 18 -- From gary.m.brown-at-exxonmobil.com Mon May 21 09:26:15 2007 8, 18 -- Received: from hoespc01.exxonmobil.com (hoespc01.exxonmobil.com [158.35.223.1]) 8, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4LEQEkv025636 8, 18 -- for {microscopy-at-microscopy.com} ; Mon, 21 May 2007 09:26:14 -0500 8, 18 -- Received: from dalnmg04.na.xom.com (dalnmg04.na.xom.com [131.126.97.123]) 8, 18 -- by hoespc01.exxonmobil.com (Switch-3.1.11/Switch-3.1.11) with ESMTP id l4LEQDBO003538 8, 18 -- for {microscopy-at-microscopy.com} ; Mon, 21 May 2007 09:26:13 -0500 (CDT) 8, 18 -- Subject: Re: Cutting tools and hazards 8, 18 -- Importance: 8, 18 -- To: microscopy-at-microscopy.com 8, 18 -- X-Mailer: Lotus Notes 652HF109 January 14, 2005 8, 18 -- Message-ID: {OF7638A511.AC313F68-ON862572E2.004EC14B-862572E2.004F4D5D-at-exxonmobil.com} 8, 18 -- From: gary.m.brown-at-exxonmobil.com 8, 18 -- Date: Mon, 21 May 2007 09:26:11 -0500 8, 18 -- X-MIMETrack: Serialize by Router on Dalnmg04.na.xom.com/S/ExxonMobil(652FP1HF193|March 8, 18 -- 02, 2006) at 05/21/2007 09:26:13 AM 8, 18 -- MIME-Version: 1.0 8, 18 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
Gary, You're not going to like this, the best safety tool is the operator. Having cut, hacked, dissected and whacked tires, hoses, belts, plastic bottles and all things rubber and polymer for over 25 years, I have only a few "guards" for you.
Rule one: before any cut is made the operator needs to look at the sample and ask: if the blade slips, where will it go? If the answer involves finger or other body parts change the configuration of the cut.
Rule two: when working under a stereomicroscope take a second to take the eyes away from the image and check the positions of the fingers, sample and blades. See rule one.
Rule three: always use sharp edges. Edges are sharp or they are not. One cut may dull an edge, but it's cheaper to replace an edge then pay emergency room bills. My worse cuts have always been with a dull knife.
Rule four: Never rush. Keep the supervisor from breathing down the operators neck, never tell a person working with a knife to hurry up.
Rule five: use the right tool for the sample. Cutting a wire reinforced hose may require mechanical saws or embedding material and a polishing wheel or a vice and a number 11 scalpel blade held by a pair of vice grips.
Rule six: always ask for assistance if you need it. Let someone else pull back a ply or component while you concentrate on the cut. Helper can were kevlar gloves and/or should use pliers, vice grips, forceps of other mechanical devices to keep there fingers out of the way.
These rules form the basic "guard" when cutting. Have I cut myself? Yes, usually when I break one of these rules.
Stay safe.............Frank
==============================Original Headers============================== 13, 18 -- From frank.karl-at-degussa.com Mon May 21 10:25:25 2007 13, 18 -- Received: from malmailout1.rz.itson.com (mailout1.degussa.com [193.100.56.173]) 13, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4LFPOZp005964 13, 18 -- for {microscopy-at-msa.microscopy.com} ; Mon, 21 May 2007 10:25:24 -0500 13, 18 -- Received: from mobuscomm01.mail.degussa.com (uss1026.applications.degussanet.com [10.88.88.98]) 13, 18 -- by malmailout1.rz.itson.com (8.13.4/8.13.4/Debian-3sarge3) with ESMTP id l4LFP0iq021693; 13, 18 -- Mon, 21 May 2007 17:25:02 +0200 13, 18 -- In-Reply-To: {200705182034.l4IKYjD2001719-at-ns.microscopy.com} 13, 18 -- Subject: Re: [Microscopy] Cutting tools and hazards 13, 18 -- To: gary.m.brown-at-exxonmobil.com, microscopy-at-msa.microscopy.com 13, 18 -- X-Mailer: Lotus Notes Release 6.5.2 June 01, 2004 13, 18 -- Message-ID: {OFB1F4AE22.691CCF4D-ON862572E2.00525BF9-852572E2.0054AFB4-at-degussa.com} 13, 18 -- From: frank.karl-at-degussa.com 13, 18 -- Date: Mon, 21 May 2007 11:24:58 -0400 13, 18 -- X-MIMETrack: Serialize by Router on MOBUSComm01/DHexternal/US(Release 6.5.1|January 21, 2004) at 13, 18 -- 05/21/2007 10:25:04 AM 13, 18 -- MIME-Version: 1.0 13, 18 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
Has anyone out there ever done electrochemical polishing on cerium? I have consulted two or three folks here with no luck. I also have Petzow's Metallographic Etching and a link to an online database of etchants. (Sorry, I cannot remember the author's name offhand and I'm pretty sure he monitors the list. Am I in trouble or what?) --
+++++++++++++++++++++++++++++
R. Ann Bliss Technologist, Chemistry Materials and Life Science Materials Science and Technology Division Lawrence Livermore National Laboratory
_____________________________
==============================Original Headers============================== 5, 16 -- From bliss5-at-llnl.gov Mon May 21 10:32:19 2007 5, 16 -- Received: from nspiron-3.llnl.gov (nspiron-3.llnl.gov [128.115.41.83]) 5, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4LFWI2A017555 5, 16 -- for {microscopy-at-sparc5.microscopy.com} ; Mon, 21 May 2007 10:32:18 -0500 5, 16 -- Received: from altaite.llnl.gov (HELO [134.9.108.10]) ([134.9.108.10]) 5, 16 -- by nspiron-3.llnl.gov with ESMTP; 21 May 2007 08:32:16 -0700 5, 16 -- X-Attachments: None 5, 16 -- X-IronPort-AV: i="4.14,561,1170662400"; 5, 16 -- d="scan'208"; a="4588966:sNHT11468682" 5, 16 -- Mime-Version: 1.0 5, 16 -- Message-Id: {p06230900c2776be04298-at-[134.9.108.10]} 5, 16 -- Date: Mon, 21 May 2007 08:32:14 -0700 5, 16 -- To: microscopy-at-ns.microscopy.com 5, 16 -- From: "R. Ann Bliss" {bliss5-at-llnl.gov} 5, 16 -- Subject: electrochemical polishing 5, 16 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
The online database you are referring to is the Metallographic Etchants Database by Henrik Kaker. It is available from South Bay Technology at
http://www.southbaytech.com/shop/med1.shtml.
I hope this helps.
Best regards-
David
David Henriks henriks-at-cox.net cell: 949-533-9100
-----Original Message----- X-from: bliss5-at-llnl.gov [mailto:bliss5-at-llnl.gov] Sent: Monday, May 21, 2007 8:38 AM To: Henriks-at-cox.net
Hello All:
Has anyone out there ever done electrochemical polishing on cerium? I have consulted two or three folks here with no luck. I also have Petzow's Metallographic Etching and a link to an online database of etchants. (Sorry, I cannot remember the author's name offhand and I'm pretty sure he monitors the list. Am I in trouble or what?) --
+++++++++++++++++++++++++++++
R. Ann Bliss Technologist, Chemistry Materials and Life Science Materials Science and Technology Division Lawrence Livermore National Laboratory
_____________________________
==============================Original Headers============================== 5, 16 -- From bliss5-at-llnl.gov Mon May 21 10:32:19 2007 5, 16 -- Received: from nspiron-3.llnl.gov (nspiron-3.llnl.gov [128.115.41.83]) 5, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4LFWI2A017555 5, 16 -- for {microscopy-at-sparc5.microscopy.com} ; Mon, 21 May 2007 10:32:18 -0500 5, 16 -- Received: from altaite.llnl.gov (HELO [134.9.108.10]) ([134.9.108.10]) 5, 16 -- by nspiron-3.llnl.gov with ESMTP; 21 May 2007 08:32:16 -0700 5, 16 -- X-Attachments: None 5, 16 -- X-IronPort-AV: i="4.14,561,1170662400"; 5, 16 -- d="scan'208"; a="4588966:sNHT11468682" 5, 16 -- Mime-Version: 1.0 5, 16 -- Message-Id: {p06230900c2776be04298-at-[134.9.108.10]} 5, 16 -- Date: Mon, 21 May 2007 08:32:14 -0700 5, 16 -- To: microscopy-at-ns.microscopy.com 5, 16 -- From: "R. Ann Bliss" {bliss5-at-llnl.gov} 5, 16 -- Subject: electrochemical polishing 5, 16 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
==============================Original Headers============================== 16, 26 -- From henriks-at-cox.net Mon May 21 11:34:17 2007 16, 26 -- Received: from fed1rmmtao104.cox.net (fed1rmmtao104.cox.net [68.230.241.42]) 16, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4LGYH5s030314 16, 26 -- for {microscopy-at-microscopy.com} ; Mon, 21 May 2007 11:34:17 -0500 16, 26 -- Received: from fed1rmimpo01.cox.net ([70.169.32.71]) 16, 26 -- by fed1rmmtao104.cox.net 16, 26 -- (InterMail vM.7.05.02.00 201-2174-114-20060621) with ESMTP 16, 26 -- id {20070521163417.SAVN15717.fed1rmmtao104.cox.net-at-fed1rmimpo01.cox.net} ; 16, 26 -- Mon, 21 May 2007 12:34:17 -0400 16, 26 -- Received: from DellE310 ([70.181.99.26]) 16, 26 -- by fed1rmimpo01.cox.net with bizsmtp 16, 26 -- id 1saC1X0040a9jum0000000; Mon, 21 May 2007 12:34:16 -0400 16, 26 -- From: "David Henriks" {henriks-at-cox.net} 16, 26 -- To: {bliss5-at-llnl.gov} 16, 26 -- Cc: {microscopy-at-microscopy.com} 16, 26 -- Subject: RE: [Microscopy] electrochemical polishing 16, 26 -- Date: Mon, 21 May 2007 09:33:52 -0700 16, 26 -- Message-ID: {008d01c79bc5$d2ded530$3c01a8c0-at-DellE310} 16, 26 -- MIME-Version: 1.0 16, 26 -- Content-Type: text/plain; 16, 26 -- charset="US-ASCII" 16, 26 -- Content-Transfer-Encoding: 7bit 16, 26 -- X-Mailer: Microsoft Office Outlook 11 16, 26 -- Thread-Index: Acebvfx1Ifm2lCTsQM+gP2jmej4S+wAByE5A 16, 26 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 16, 26 -- In-Reply-To: AAAAAMY7gB18BfNPgPDRzX6JX+Ek6SYA ==============================End of - Headers==============================
Please the link http://www.kaker.com/etch/demo/index.html.
Henrik
henriks-at-cox.net wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Ann: } } The online database you are referring to is the Metallographic Etchants } Database by Henrik Kaker. It is available from South Bay Technology at } } http://www.southbaytech.com/shop/med1.shtml. } } I hope this helps. } } Best regards- } } David } } David Henriks } henriks-at-cox.net } cell: 949-533-9100 } } -----Original Message----- } X-from: bliss5-at-llnl.gov [mailto:bliss5-at-llnl.gov] } Sent: Monday, May 21, 2007 8:38 AM } To: Henriks-at-cox.net } Subject: [Microscopy] electrochemical polishing } } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Hello All: } } Has anyone out there ever done electrochemical polishing on cerium? I } have consulted two or three folks here with no luck. I also have } Petzow's Metallographic Etching and a link to an online database of } etchants. (Sorry, I cannot remember the author's name offhand and I'm } pretty sure he monitors the list. Am I in trouble or what?) }
-- Henrik Kaker Ph.D. Metal Ravne d.o.o. SEM-EDS Laboratory Koroska cesta 14 SI-2390 Ravne Slovenia Phone: +386 2 870 7076 GSM: +386 31 380 875 http://www2.arnes.si/~sgszmera1/index.html
==============================Original Headers============================== 7, 26 -- From Henrik.Kaker-at-guest.arnes.si Mon May 21 13:50:25 2007 7, 26 -- Received: from avs3.arnes.si (avs3.arnes.si [193.2.1.68]) 7, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4LIoOmr012234 7, 26 -- for {microscopy-at-microscopy.com} ; Mon, 21 May 2007 13:50:25 -0500 7, 26 -- Received: from localhost (avs3.arnes.si [193.2.1.68]) 7, 26 -- by avs3.arnes.si (Postfix) with ESMTP id D66DD1D79B3 7, 26 -- for {microscopy-at-microscopy.com} ; Mon, 21 May 2007 20:50:23 +0200 (CEST) 7, 26 -- Received: from avs3.arnes.si ([193.2.1.68]) 7, 26 -- by localhost (avs3.arnes.si [193.2.1.68]) (amavisd-new, port 10024) 7, 26 -- with ESMTP id 58765-07 for {microscopy-at-microscopy.com} ; 7, 26 -- Mon, 21 May 2007 20:50:23 +0200 (CEST) 7, 26 -- Received: from [89.212.22.21] (89-212-22-21.static.dsl.t-2.net [89.212.22.21]) 7, 26 -- by avs3.arnes.si (Postfix) with ESMTP id 8006F1D79A5 7, 26 -- for {microscopy-at-microscopy.com} ; Mon, 21 May 2007 20:50:23 +0200 (CEST) 7, 26 -- Message-ID: {4651E9EF.6050309-at-guest.arnes.si} 7, 26 -- Date: Mon, 21 May 2007 20:50:23 +0200 7, 26 -- From: Henrik Kaker {Henrik.Kaker-at-guest.arnes.si} 7, 26 -- User-Agent: Thunderbird 2.0.0.0 (Windows/20070326) 7, 26 -- MIME-Version: 1.0 7, 26 -- To: microscopy-at-microscopy.com 7, 26 -- Subject: Re: [Microscopy] RE: electrochemical polishing 7, 26 -- References: {200705211634.l4LGYZDu030689-at-ns.microscopy.com} 7, 26 -- In-Reply-To: {200705211634.l4LGYZDu030689-at-ns.microscopy.com} 7, 26 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 7, 26 -- Content-Transfer-Encoding: 7bit 7, 26 -- X-Virus-Scanned: by amavisd-new at arnes.si ==============================End of - Headers==============================
A few of my colleagues are trying to study transport behaviour of lipid bilayer membranes. This requires a membrane to be deposited over a (tiny) hole between two aqeous compartments. The (in)stability of the suspended part of such a membrane is a major complication. I am wondering whether a support technique from the microscopy field might be helpful here. In particular I am curious about the possibility of using a holey carbon-like approach. Not being a holey-carbon expert myself, I would much appreciate if anyone could answer the questions underneath or provide similar or alternative suggestions.
1. What materials are typically used to make holey or lacey films. Just amorphous carbon and formvar?
2. What are the typical dimensions attainable for holey or lacey films, in terms of film thickness and hole size. For our purposes, 20-40 nm hole diameter would be ideal, but 100 - 200 nm would be very helpful already.
3. Are there good descriptions of the preparation or availability of such holey films?
4. Does anyone have experience in supporting lipid bilayers in different ways, e.g. for TEM investigation?
Thank you all for your help!
With kind regards,
Coen van den Brom
----------------------------
dr CR van den Brom Department of Nanoscale Chemistry School of Chemistry University of Birmingham Edgbaston Birmingham B15 2TT United Kingdom
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One very important rule that I always check: make sure the sharp side is down! Sounds stupid, but I can't tell you how many times people have cut themselves because the sharp edge was up! So, to prevent such disasters, I always ask "Is the sharp side down?" Maybe it's just that I'm getting older and catch myself every now and then with the sharp side up. And I will strongly second the rule about not rushing or allowing other distraction (conversations, etc)--when I have hurt myself the worst it has been someone standing there yammering at me.
Roger Moretz
On 5/21/07, frank.karl-at-degussa.com {frank.karl-at-degussa.com} wrote: } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Gary, } You're not going to like this, the best safety tool is the operator. } Having cut, hacked, dissected and whacked tires, hoses, belts, plastic } bottles and all things rubber and polymer for over 25 years, I have only a } few "guards" for you. } } Rule one: before any cut is made the operator needs to look at the sample } and ask: if the blade slips, where will it go? If the answer involves } finger or other body parts change the configuration of the cut. } } Rule two: when working under a stereomicroscope take a second to take the } eyes away from the image and check the positions of the fingers, sample and } blades. See rule one. } } Rule three: always use sharp edges. Edges are sharp or they are not. One } cut may dull an edge, but it's cheaper to replace an edge then pay } emergency room bills. My worse cuts have always been with a dull knife. } } Rule four: Never rush. Keep the supervisor from breathing down the } operators neck, never tell a person working with a knife to hurry up. } } Rule five: use the right tool for the sample. Cutting a wire reinforced } hose may require mechanical saws or embedding material and a polishing } wheel or a vice and a number 11 scalpel blade held by a pair of vice grips. } } Rule six: always ask for assistance if you need it. Let someone else pull } back a ply or component while you concentrate on the cut. Helper can were } kevlar gloves and/or should use pliers, vice grips, forceps of other } mechanical devices to keep there fingers out of the way. } } These rules form the basic "guard" when cutting. Have I cut myself? Yes, } usually when I break one of these rules. } } Stay safe.............Frank } } } } } } ==============================Original Headers============================== } 13, 18 -- From frank.karl-at-degussa.com Mon May 21 10:25:25 2007 } 13, 18 -- Received: from malmailout1.rz.itson.com (mailout1.degussa.com [193.100.56.173]) } 13, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4LFPOZp005964 } 13, 18 -- for {microscopy-at-msa.microscopy.com} ; Mon, 21 May 2007 10:25:24 -0500 } 13, 18 -- Received: from mobuscomm01.mail.degussa.com (uss1026.applications.degussanet.com [10.88.88.98]) } 13, 18 -- by malmailout1.rz.itson.com (8.13.4/8.13.4/Debian-3sarge3) with ESMTP id l4LFP0iq021693; } 13, 18 -- Mon, 21 May 2007 17:25:02 +0200 } 13, 18 -- In-Reply-To: {200705182034.l4IKYjD2001719-at-ns.microscopy.com} } 13, 18 -- Subject: Re: [Microscopy] Cutting tools and hazards } 13, 18 -- To: gary.m.brown-at-exxonmobil.com, microscopy-at-msa.microscopy.com } 13, 18 -- X-Mailer: Lotus Notes Release 6.5.2 June 01, 2004 } 13, 18 -- Message-ID: {OFB1F4AE22.691CCF4D-ON862572E2.00525BF9-852572E2.0054AFB4-at-degussa.com} } 13, 18 -- From: frank.karl-at-degussa.com } 13, 18 -- Date: Mon, 21 May 2007 11:24:58 -0400 } 13, 18 -- X-MIMETrack: Serialize by Router on MOBUSComm01/DHexternal/US(Release 6.5.1|January 21, 2004) at } 13, 18 -- 05/21/2007 10:25:04 AM } 13, 18 -- MIME-Version: 1.0 } 13, 18 -- Content-type: text/plain; charset=US-ASCII } ==============================End of - Headers============================== }
==============================Original Headers============================== 3, 28 -- From rcmoretz-at-gmail.com Mon May 21 21:28:04 2007 3, 28 -- Received: from nz-out-0506.google.com (nz-out-0506.google.com [64.233.162.235]) 3, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4M2S3MO001852 3, 28 -- for {Microscopy-at-microscopy.com} ; Mon, 21 May 2007 21:28:04 -0500 3, 28 -- Received: by nz-out-0506.google.com with SMTP id x3so2332215nzd 3, 28 -- for {Microscopy-at-microscopy.com} ; Mon, 21 May 2007 19:28:03 -0700 (PDT) 3, 28 -- DKIM-Signature: a=rsa-sha1; c=relaxed/relaxed; 3, 28 -- d=gmail.com; s=beta; 3, 28 -- h=domainkey-signature:received:received:message-id:date:from:to:subject:in-reply-to:mime-version:content-type:content-transfer-encoding:content-disposition:references; 3, 28 -- b=iXHSC7qV1SDQKecEn7SZ32Wv/STWm6AuJm/x4Jf2cTU4gJeS68V0cyxoPCaSU2VG76y+qD7bkREYqf1TWGBcw77NK0cbbJvIcSAReW26d/zlzpRvWJ9sF+K2ZyCwY2E0wErQNr7DVGFWEVKumcnRb4dCNRiDx9XtdTP7vRpAQlE= 3, 28 -- DomainKey-Signature: a=rsa-sha1; c=nofws; 3, 28 -- d=gmail.com; s=beta; 3, 28 -- h=received:message-id:date:from:to:subject:in-reply-to:mime-version:content-type:content-transfer-encoding:content-disposition:references; 3, 28 -- b=sczgI7ze1OBOgTPsfFfVUrfXKao8NbJJXcgqpu8ccgb8iRxe9Qk8flr+4ZCrkG3yxISd8tHNxUEGet3N7pyoCoC0hvZTGhXj16MsJyhRZGFiRbF5nB+yvSGdezpkAVQEA6gQHHfaAbud8tjrse0lbTQI8v0nbsMMWj9sfQEsGf0= 3, 28 -- Received: by 10.65.105.3 with SMTP id h3mr12133361qbm.1179800883370; 3, 28 -- Mon, 21 May 2007 19:28:03 -0700 (PDT) 3, 28 -- Received: by 10.64.193.8 with HTTP; Mon, 21 May 2007 19:28:03 -0700 (PDT) 3, 28 -- Message-ID: {950e3cfd0705211928m195170f7o95d135e6ff4c09e5-at-mail.gmail.com} 3, 28 -- Date: Mon, 21 May 2007 22:28:03 -0400 3, 28 -- From: "Roger Moretz" {rcmoretz-at-gmail.com} 3, 28 -- To: frank.karl-at-degussa.com, "Microscopy Listserv" {Microscopy-at-microscopy.com} 3, 28 -- Subject: Re: [Microscopy] Re: Cutting tools and hazards 3, 28 -- In-Reply-To: {200705211529.l4LFTuvv013962-at-ns.microscopy.com} 3, 28 -- MIME-Version: 1.0 3, 28 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 3, 28 -- Content-Transfer-Encoding: 7bit 3, 28 -- Content-Disposition: inline 3, 28 -- References: {200705211529.l4LFTuvv013962-at-ns.microscopy.com} ==============================End of - Headers==============================
I am participating in a workshop (again) where they want me to train 14 people to find GFP in Arabidopsis in both confocal and TEM. I have tried fixing young leaves and cotyledons (cut into {1mm strips) in 2% paraformaldehyde/0.1% glutaraldehyde in 0.1M Na cacodylate with 2mM CaCl2, washing, dehydrating, embedding in LR White, polymerized at 50C. I'm getting holes in the resin (I didn't particularly rush through dehydration, but I can prolong it, and I did not pull a vacuum, but I can), and I'm not seeing any GFP in 1 um sections in the confocal. I am reluctant to keep going with these samples and then try to do anti-GFP for TEM if I'm not seeing GFP (but the organizers of the workshop don't seem to care, as long as we go through the motions)(Sigh). I would be grateful for any tips about any step along the way from anybody!
Mahalo! Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
==============================Original Headers============================== 5, 19 -- From tina-at-pbrc.hawaii.edu Mon May 21 22:24:21 2007 5, 19 -- Received: from halia.pbrc.hawaii.edu (halia.pbrc.hawaii.edu [128.171.22.7]) 5, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4M3OKBV014227 5, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 21 May 2007 22:24:21 -0500 5, 19 -- Received: from halia.pbrc.hawaii.edu (localhost [127.0.0.1]) 5, 19 -- by halia.pbrc.hawaii.edu (8.12.11/8.12.11) with ESMTP id l4M3OFpZ013256 5, 19 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 5, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 21 May 2007 17:24:17 -1000 (HST) 5, 19 -- Received: from localhost by halia.pbrc.hawaii.edu (8.12.11/8.12.11/Submit) with ESMTP id l4M3OFsM013253 5, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 21 May 2007 17:24:15 -1000 (HST) 5, 19 -- X-Authentication-Warning: halia.pbrc.hawaii.edu: tina owned process doing -bs 5, 19 -- Date: Mon, 21 May 2007 17:24:14 -1000 (HST) 5, 19 -- From: Tina Carvalho {tina-at-pbrc.hawaii.edu} 5, 19 -- X-Sender: tina-at-halia 5, 19 -- To: Microscopy Listserver {Microscopy-at-MSA.Microscopy.Com} 5, 19 -- Subject: Arabidopsis GFP in TEM 5, 19 -- Message-ID: {Pine.GSO.4.21.0705211711040.13144-100000-at-halia} 5, 19 -- MIME-Version: 1.0 5, 19 -- Content-Type: TEXT/PLAIN; charset=US-ASCII ==============================End of - Headers==============================
Roger Moretz wrote: =============================================================== One very important rule that I always check: make sure the sharp side is down! Sounds stupid, but I can't tell you how many times people have cut themselves because the sharp edge was up! So, to prevent such disasters, I always ask "Is the sharp side down?" Maybe it's just that I'm getting older and catch myself every now and then with the sharp side up. And I will strongly second the rule about not rushing or allowing other distraction (conversations, etc)--when I have hurt myself the worst it has been someone standing there yammering at me. =============================================================== In our laboratory and production facilities, we do not permit the use of (the cheaper) double sided razor blades because of the reasons cited above. A single edge razor blade (we prefer the GEM type, see URL http://www.2spi.com/catalog/tools/smtol14.shtml is all that is permitted.
People would probably be amazed at the number of eye injuries that occur (not necessarily in EM labs) when one uses a razor (or scalpel) blade (of any type) not so much for cutting but for "lifting" or "prying", now the forces are not within the plane of the blade but are perpendicular to it, and as a result of the shear stresses against which the blade was never designed, a corner of the blade can easily "fly" off at high speed and injure the worker if not wearing eye protection. This is also a good reason why eye protection should be worn by anyone in the vicinity of anyone using any kind of a razor blade.
Over the years, we have been asked to do quite a number of failure analysis studies and reports on such blade fragments that caused eye injury (or even blindness). I don't recall that we have ever found a "material defect" in such studies, but there was apparently more than enough evidence to suggest that the user was improperly using the blade (in shear).
I also agree with Roger that idle "chit-chat" can be hazardous when one is trying to do work with potentially dangerous tools at the same time.
Disclaimer: SPI Supplies distributes the GEM single edge "scientific" razor blades so we would have a vested interest in seeing more of our customers using this type of blade.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
==============================Original Headers============================== 10, 20 -- From cgarber-at-2spi.com Tue May 22 02:20:20 2007 10, 20 -- Received: from diskless5.axs2000.net (mail.axs2000.net [209.120.196.43]) 10, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4M7KKJe002709 10, 20 -- for {microscopy-at-msa.microscopy.com} ; Tue, 22 May 2007 02:20:20 -0500 10, 20 -- Received: from [10.104.63.56] (62-50-216-20.client.stsn.net [62.50.216.20]) 10, 20 -- (authenticated bits=0) 10, 20 -- by diskless5.axs2000.net (8.12.11/8.12.11) with ESMTP id l4M7KIEB032151 10, 20 -- for {microscopy-at-msa.microscopy.com} ; Tue, 22 May 2007 03:20:19 -0400 10, 20 -- X-IDV-FirstRcvd: 62-50-216-20.client.stsn.net [62.50.216.20] 10, 20 -- X-IDV-HELO: [10.104.63.56] 10, 20 -- X-IDV-Authenticated-User: cgarber 10, 20 -- Message-ID: {465299A2.7010808-at-2spi.com} 10, 20 -- Date: Tue, 22 May 2007 03:20:02 -0400 10, 20 -- From: "Garber, Charles A" {cgarber-at-2spi.com} 10, 20 -- User-Agent: Thunderbird 2.0.0.0 (Windows/20070326) 10, 20 -- MIME-Version: 1.0 10, 20 -- To: Microscopy Listserver {microscopy-at-msa.microscopy.com} 10, 20 -- Subject: Cutting tools and hazards 10, 20 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 10, 20 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Here are some basic questions from the basic SEM user I am: I would like to visualize salt crystals by SEM and make EDX analysis if possible. I just leave a drop of salt solution on a carbon tab dry, that's all! My questions:
- Do I need to carbon-coat the crystals? - Is there a risk to evaporate the crystals under the electron beam and make the column dirty? (I usually don't use HV higher than 15 kV). Would carbon coating help dissipate the heat in this regard?
Now another question related to SEM but not to crystals: When I analyse fine particles by EDX, I never know if I analyse only the particle or also part of the material which is behind the particle. Is there a way to determine the depth of analysis of the material studied? I suppose that it depends on the HV and on the nature of the material itself, but I already would be happy to have a order of magnitude. Is this 10 nm or 2 µm deep? Lets say I analyze at 15kV (1) NaCl crystals (2) Quartz (3) Feldpar (4) Copper, what would be the expected "depth of analysis" in these samples? Does it depend on the size of the beam?
Best regards,
Stephane
____________________________________________________________________________________ Need Mail bonding? Go to the Yahoo! Mail Q&A for great tips from Yahoo! Answers users. http://answers.yahoo.com/dir/?link=list&sid=396546091
==============================Original Headers============================== 13, 19 -- From nizets2-at-yahoo.com Tue May 22 04:46:12 2007 13, 19 -- Received: from web37405.mail.mud.yahoo.com (web37405.mail.mud.yahoo.com [209.191.91.137]) 13, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l4M9kCVc017352 13, 19 -- for {microscopy-at-microscopy.com} ; Tue, 22 May 2007 04:46:12 -0500 13, 19 -- Received: (qmail 54717 invoked by uid 60001); 22 May 2007 09:46:08 -0000 13, 19 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 13, 19 -- s=s1024; d=yahoo.com; 13, 19 -- h=X-YMail-OSG:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding:Message-ID; 13, 19 -- b=zBbqWoaMZ9PlXpKnS7/04OoVZZIK8DFKFHcyjE3Fb/m3Zl4auB81Rs10Qpyy2eM+Nm+m0B7lAIn+OTWrzWZNMScRq4hkKwM1at2lB+Ok+wl3FvDKesHuEIr4DQMXBSph2qIICP+SmycYwMy8nLABeHvgi3DVBD88TCbeU3lTI7w=; 13, 19 -- X-YMail-OSG: _Sy6fjQVM1n_aA9U2sOeO5UGkSLDD5RhnQ74_ILWjiw02eS6PDlVt_Y7dn7dCMkDiw-- 13, 19 -- Received: from [80.122.101.102] by web37405.mail.mud.yahoo.com via HTTP; Tue, 22 May 2007 02:46:08 PDT 13, 19 -- Date: Tue, 22 May 2007 02:46:08 -0700 (PDT) 13, 19 -- From: Stephane Nizet {nizets2-at-yahoo.com} 13, 19 -- Subject: basic SEM: imaging salt crystals 13, 19 -- To: microscopy-at-microscopy.com 13, 19 -- MIME-Version: 1.0 13, 19 -- Content-Type: text/plain; charset=iso-8859-1 13, 19 -- Content-Transfer-Encoding: 8bit 13, 19 -- Message-ID: {210050.53719.qm-at-web37405.mail.mud.yahoo.com} ==============================End of - Headers==============================
I completely share this opinion. Seriousness and concentration are not an art, it is available to anybody wanting them. Correct training is also most important. I met a girl who embedded her Epon in eppendorf tubes. Then she took the tube in one hand (bare hands!) and cut through the plastic with the other hand using a razor blade! I mean, you don't need to be a genius to expect an accident one day or another.
I don't cut blocks for 25 years, but still for 15 good years and I never had the smallest accident. I don't think it is luck. Now I understand that one cannot always control what others do. In this case ask the person to wear plexiglas goggles, bullet-proof vest, kevlar gloves and there is still the risk that he cuts his ear!
Frank's rules are excellent, but most of them could be all replaced by one only rule: use your common sense. Rules 3 and 4 are critical and should be part of a correct training.
Best regards, Stephane
--- frank.karl-at-degussa.com wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Gary, } You're not going to like this, the best safety tool } is the operator. } Having cut, hacked, dissected and whacked tires, } hoses, belts, plastic } bottles and all things rubber and polymer for over } 25 years, I have only a } few "guards" for you. } } Rule one: before any cut is made the operator needs } to look at the sample } and ask: if the blade slips, where will it go? If } the answer involves } finger or other body parts change the configuration } of the cut. } } Rule two: when working under a stereomicroscope take } a second to take the } eyes away from the image and check the positions of } the fingers, sample and } blades. See rule one. } } Rule three: always use sharp edges. Edges are sharp } or they are not. One } cut may dull an edge, but it's cheaper to replace an } edge then pay } emergency room bills. My worse cuts have always been } with a dull knife. } } Rule four: Never rush. Keep the supervisor from } breathing down the } operators neck, never tell a person working with a } knife to hurry up. } } Rule five: use the right tool for the sample. } Cutting a wire reinforced } hose may require mechanical saws or embedding } material and a polishing } wheel or a vice and a number 11 scalpel blade held } by a pair of vice grips. } } Rule six: always ask for assistance if you need it. } Let someone else pull } back a ply or component while you concentrate on the } cut. Helper can were } kevlar gloves and/or should use pliers, vice grips, } forceps of other } mechanical devices to keep there fingers out of the } way. } } These rules form the basic "guard" when cutting. } Have I cut myself? Yes, } usually when I break one of these rules. } } Stay safe.............Frank } } } } } } ==============================Original } Headers============================== } 13, 18 -- From frank.karl-at-degussa.com Mon May 21 } 10:25:25 2007 } 13, 18 -- Received: from malmailout1.rz.itson.com } (mailout1.degussa.com [193.100.56.173]) } 13, 18 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with ESMTP id } l4LFPOZp005964 } 13, 18 -- for {microscopy-at-msa.microscopy.com} ; Mon, } 21 May 2007 10:25:24 -0500 } 13, 18 -- Received: from } mobuscomm01.mail.degussa.com } (uss1026.applications.degussanet.com [10.88.88.98]) } 13, 18 -- by malmailout1.rz.itson.com } (8.13.4/8.13.4/Debian-3sarge3) with ESMTP id } l4LFP0iq021693; } 13, 18 -- Mon, 21 May 2007 17:25:02 +0200 } 13, 18 -- In-Reply-To: } {200705182034.l4IKYjD2001719-at-ns.microscopy.com} } 13, 18 -- Subject: Re: [Microscopy] Cutting tools } and hazards } 13, 18 -- To: gary.m.brown-at-exxonmobil.com, } microscopy-at-msa.microscopy.com } 13, 18 -- X-Mailer: Lotus Notes Release 6.5.2 June } 01, 2004 } 13, 18 -- Message-ID: } {OFB1F4AE22.691CCF4D-ON862572E2.00525BF9-852572E2.0054AFB4-at-degussa.com} } 13, 18 -- From: frank.karl-at-degussa.com } 13, 18 -- Date: Mon, 21 May 2007 11:24:58 -0400 } 13, 18 -- X-MIMETrack: Serialize by Router on } MOBUSComm01/DHexternal/US(Release 6.5.1|January 21, } 2004) at } 13, 18 -- 05/21/2007 10:25:04 AM } 13, 18 -- MIME-Version: 1.0 } 13, 18 -- Content-type: text/plain; charset=US-ASCII } ==============================End of - } Headers============================== }
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==============================Original Headers============================== 11, 20 -- From nizets2-at-yahoo.com Tue May 22 07:09:31 2007 11, 20 -- Received: from web37415.mail.mud.yahoo.com (web37415.mail.mud.yahoo.com [209.191.91.147]) 11, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l4MC9UnH031630 11, 20 -- for {microscopy-at-microscopy.com} ; Tue, 22 May 2007 07:09:30 -0500 11, 20 -- Received: (qmail 85899 invoked by uid 60001); 22 May 2007 12:09:30 -0000 11, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 11, 20 -- s=s1024; d=yahoo.com; 11, 20 -- h=X-YMail-OSG:Received:Date:From:Subject:To:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding:Message-ID; 11, 20 -- b=mXZmtxIucOpqqdN5VfixsyF8X6jCeqZQnRwKvWT8uINLSI2HxWL3+9FWVajtuWOqLL7c6b0X6/+2TH1BS6B08lsbRZjV6OojIz/24PqTwyV4icfg0kofzqdU87mnqlcP4n8e2rPZgMjvO0hj6lkFmzO7YLxx88vbDOIIxHZ/irg=; 11, 20 -- X-YMail-OSG: 9V3DWqYVM1kva3b0Djsr_XdCG2uR1eQqWVdMdvOCP3Iidqe3KAAXGcs3CKPa2xdLgXO7qDgbeda18XmM0DxkBEHQNIg689BQJVML4J76eBmHYAiio2zTRpvoQYeG9WXjwXRQjshG6jxOOuJl3O1.v_ZhEXCAnS87zqH8J6LIhV7dzwPj17nPc1VfCOUwVxURSYJJn16Nl5Bk7zlIDg-- 11, 20 -- Received: from [80.122.101.102] by web37415.mail.mud.yahoo.com via HTTP; Tue, 22 May 2007 05:09:30 PDT 11, 20 -- Date: Tue, 22 May 2007 05:09:30 -0700 (PDT) 11, 20 -- From: Stephane Nizet {nizets2-at-yahoo.com} 11, 20 -- Subject: Re: [Microscopy] Re: Cutting tools and hazards 11, 20 -- To: microscopy-at-microscopy.com 11, 20 -- In-Reply-To: {200705211529.l4LFTmh2013672-at-ns.microscopy.com} 11, 20 -- MIME-Version: 1.0 11, 20 -- Content-Type: text/plain; charset=iso-8859-1 11, 20 -- Content-Transfer-Encoding: 8bit 11, 20 -- Message-ID: {603281.84638.qm-at-web37415.mail.mud.yahoo.com} ==============================End of - Headers==============================
300 ml/min. Perfuse either through the carotid artery or left ventricle (occlude descending aorta).
Best regards, Walter F. Bobrowski Sr. Scientist Pfizer Global R&D 2800 Plymouth Rd. Ann Arbor, MI 48105
TEL: 734-622-7814 FAX: 734-622-5718
"Like the strength of a steel rod, the true character of a person can only be known under extreme stress." - Leslie Fieger
-----Original Message----- X-from: mpease-at-jhmi.edu [mailto:mpease-at-jhmi.edu] Sent: Monday, May 21, 2007 7:25 PM To: Bobrowski, Walter
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==============================Original Headers============================== 7, 11 -- From zaluzec-at-microscopy.com Mon May 21 18:19:37 2007 7, 11 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) 7, 11 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4LNJbJx007966 7, 11 -- for {microscopy-at-microscopy.com} ; Mon, 21 May 2007 18:19:37 -0500 7, 11 -- Mime-Version: 1.0 7, 11 -- Message-Id: {p06240806c277d979e099-at-[206.69.208.22]} 7, 11 -- Date: Mon, 21 May 2007 18:19:36 -0500 7, 11 -- To: microscopy-at-microscopy.com 7, 11 -- From: mpease-at-jhmi.edu (by way of MicroscopyListserver) 7, 11 -- Subject: viaWWW: monkey perfusion methodology 7, 11 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
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==============================Original Headers============================== 20, 31 -- From Walter.Bobrowski-at-pfizer.com Tue May 22 07:27:56 2007 20, 31 -- Received: from gromsgom01.pfizer.com (gromsgo.pfizer.com [148.168.224.84]) 20, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4MCRtXc011019 20, 31 -- for {microscopy-at-microscopy.com} ; Tue, 22 May 2007 07:27:56 -0500 20, 31 -- Received: from mopamrexc01.amer.pfizer.com (mopamrexc01.pfizer.com [170.116.32.254]) 20, 31 -- by gromsgom01.pfizer.com (8.13.7/8.13.7) with ESMTP id l4MCRYAY010379; 20, 31 -- Tue, 22 May 2007 08:27:50 -0400 20, 31 -- Received: from mopamrexc03.amer.pfizer.com ([170.116.30.69]) by mopamrexc01.amer.pfizer.com with Microsoft SMTPSVC(6.0.3790.1830); 20, 31 -- Tue, 22 May 2007 08:27:46 -0400 20, 31 -- Received: from anaamrexm01.amer.pfizer.com ([162.48.101.10]) by mopamrexc03.amer.pfizer.com with Microsoft SMTPSVC(6.0.3790.1830); 20, 31 -- Tue, 22 May 2007 08:27:46 -0400 20, 31 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 20, 31 -- Content-class: urn:content-classes:message 20, 31 -- MIME-Version: 1.0 20, 31 -- Content-Type: text/plain; 20, 31 -- charset="us-ascii" 20, 31 -- Subject: RE: [Microscopy] viaWWW: monkey perfusion methodology 20, 31 -- Date: Tue, 22 May 2007 08:27:39 -0400 20, 31 -- Message-ID: {CF89AF6ABD93B746A55BCCD9480A4B3D09227763-at-anaamrexm01.amer.pfizer.com} 20, 31 -- In-Reply-To: {200705212324.l4LNOv2b016731-at-ns.microscopy.com} 20, 31 -- X-MS-Has-Attach: 20, 31 -- X-MS-TNEF-Correlator: 20, 31 -- Thread-Topic: [Microscopy] viaWWW: monkey perfusion methodology 20, 31 -- Thread-Index: Aceb/z/rFaL3idT1TMqHjz+q29pclAAbRiog 20, 31 -- From: "Bobrowski, Walter" {Walter.Bobrowski-at-pfizer.com} 20, 31 -- To: {mpease-at-jhmi.edu} , {microscopy-at-microscopy.com} 20, 31 -- X-OriginalArrivalTime: 22 May 2007 12:27:46.0092 (UTC) FILETIME=[9A3516C0:01C79C6C] 20, 31 -- X-Proofpoint-Virus-Version: vendor=fsecure engine=4.65.5502:2.3.11,1.2.37,4.0.164 definitions=2007-05-22_05:2007-05-22,2007-05-22,2007-05-22 signatures=0 20, 31 -- X-Proofpoint-Spam-Reason: safe 20, 31 -- Content-Transfer-Encoding: 8bit 20, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4MCRtXc011019 ==============================End of - Headers==============================
Can you screen for GFP by fluorescence after fixation, then dehydration, and finally embedding to see where the signal is lost?
You may have better luck using a water soluble resin such as Durcupan. I have not tried this resin for ICC but it is relatively soft so may work. Advantage is that you do not have to dehydrate in ETOH or acetone and this may help preserve the GFP signal.
Debby
Debby Sherman, Director Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907
} From: {tina-at-pbrc.hawaii.edu} } Reply-To: {tina-at-pbrc.hawaii.edu} } Date: Mon, 21 May 2007 22:26:18 -0500 } To: {dsherman-at-purdue.edu} } Subject: [Microscopy] Arabidopsis GFP in TEM } } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Hi, All- } } I am participating in a workshop (again) where they want me to train 14 } people to find GFP in Arabidopsis in both confocal and TEM. I have tried } fixing young leaves and cotyledons (cut into {1mm strips) in 2% } paraformaldehyde/0.1% glutaraldehyde in 0.1M Na cacodylate with 2mM CaCl2, } washing, dehydrating, embedding in LR White, polymerized at 50C. I'm } getting holes in the resin (I didn't particularly rush through } dehydration, but I can prolong it, and I did not pull a vacuum, but I } can), and I'm not seeing any GFP in 1 um sections in the confocal. I am } reluctant to keep going with these samples and then try to do anti-GFP for } TEM if I'm not seeing GFP (but the organizers of the workshop don't seem } to care, as long as we go through the motions)(Sigh). I would be grateful } for any tips about any step along the way from anybody! } } Mahalo! } Tina } } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } **************************************************************************** } } } ==============================Original Headers============================== } 5, 19 -- From tina-at-pbrc.hawaii.edu Mon May 21 22:24:21 2007 } 5, 19 -- Received: from halia.pbrc.hawaii.edu (halia.pbrc.hawaii.edu } [128.171.22.7]) } 5, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } l4M3OKBV014227 } 5, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 21 May 2007 22:24:21 -0500 } 5, 19 -- Received: from halia.pbrc.hawaii.edu (localhost [127.0.0.1]) } 5, 19 -- by halia.pbrc.hawaii.edu (8.12.11/8.12.11) with ESMTP id } l4M3OFpZ013256 } 5, 19 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) } 5, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 21 May 2007 17:24:17 -1000 } (HST) } 5, 19 -- Received: from localhost by halia.pbrc.hawaii.edu } (8.12.11/8.12.11/Submit) with ESMTP id l4M3OFsM013253 } 5, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 21 May 2007 17:24:15 -1000 } (HST) } 5, 19 -- X-Authentication-Warning: halia.pbrc.hawaii.edu: tina owned process } doing -bs } 5, 19 -- Date: Mon, 21 May 2007 17:24:14 -1000 (HST) } 5, 19 -- From: Tina Carvalho {tina-at-pbrc.hawaii.edu} } 5, 19 -- X-Sender: tina-at-halia } 5, 19 -- To: Microscopy Listserver {Microscopy-at-MSA.Microscopy.Com} } 5, 19 -- Subject: Arabidopsis GFP in TEM } 5, 19 -- Message-ID: {Pine.GSO.4.21.0705211711040.13144-100000-at-halia} } 5, 19 -- MIME-Version: 1.0 } 5, 19 -- Content-Type: TEXT/PLAIN; charset=US-ASCII } ==============================End of - Headers==============================
==============================Original Headers============================== 9, 23 -- From dsherman-at-purdue.edu Tue May 22 07:35:10 2007 9, 23 -- Received: from 1061exfe01a.itap.purdue.edu (1061exfe01a.itap.purdue.edu [128.210.1.8]) 9, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4MCZAOj022639 9, 23 -- for {microscopy-at-microscopy.com} ; Tue, 22 May 2007 07:35:10 -0500 9, 23 -- Received: from exchange.purdue.edu ([172.21.6.23]) by 1061exfe01a.itap.purdue.edu with Microsoft SMTPSVC(6.0.3790.3959); 9, 23 -- Tue, 22 May 2007 08:35:10 -0400 9, 23 -- Received: from 128.210.161.94 ([128.210.161.94]) by EXCH04.purdue.lcl ([172.21.6.26]) via Exchange Front-End Server exch.itap.purdue.edu ([128.210.63.104]) with Microsoft Exchange Server HTTP-DAV ; 9, 23 -- Tue, 22 May 2007 12:35:09 +0000 9, 23 -- User-Agent: Microsoft-Entourage/11.3.3.061214 9, 23 -- Date: Tue, 22 May 2007 08:35:08 -0400 9, 23 -- Subject: Re: [Microscopy] Arabidopsis GFP in TEM 9, 23 -- From: Debby Sherman {dsherman-at-purdue.edu} 9, 23 -- To: Tina Carvalho {tina-at-pbrc.hawaii.edu} , 9, 23 -- "message to: MSA list" {microscopy-at-microscopy.com} 9, 23 -- Message-ID: {C2785BBC.1C71C%dsherman-at-purdue.edu} 9, 23 -- Thread-Topic: [Microscopy] Arabidopsis GFP in TEM 9, 23 -- Thread-Index: AcecbaGa4FnM7QhgEdyyHwARJN08Mg== 9, 23 -- In-Reply-To: {200705220326.l4M3QIa9017954-at-ns.microscopy.com} 9, 23 -- Mime-version: 1.0 9, 23 -- Content-type: text/plain; 9, 23 -- charset="US-ASCII" 9, 23 -- Content-transfer-encoding: 7bit 9, 23 -- X-OriginalArrivalTime: 22 May 2007 12:35:10.0024 (UTC) FILETIME=[A2CFBC80:01C79C6D] ==============================End of - Headers==============================
0) Purchase a copy of Electron Flight Simulator or download a copy of WinXray (from McGill). You should run simulations to estimate your excitation volume.
1) Personally, I would not put a drop of liquid on carbon tabs or double sided tape. It is bad practice, as the water does interact with the adhesive.
Instead, use a silicon wafer (you can purchase precut wafers), carbon planchet, Be planchet, etc....
2) If the substrates are conducting (as are the ones mentioned), then you will not need much overcoating. Personally, a few nm of gold on an incline sample will do the trick.
regards,
Jim
PS: OoO away...............
} From mail-at-ns.microscopy.com Tue May 22 05:45:52 2007 } Return-Path: {mail-at-ns.microscopy.com} } Received: from ns.microscopy.com (microscopy.com [206.69.208.10]) } by www.matscieng.sunysb.edu (8.11.6/8.11.6) with ESMTP id l4M9jqJ16923 } for {jquinn-at-www.matscieng.sunysb.edu} ; Tue, 22 May 2007 05:45:52 -0400 } Received: from ns.microscopy.com (localhost.localdomain [127.0.0.1]) } by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4M9l99O018527 } for {jquinn-at-www.matscieng.sunysb.edu} ; Tue, 22 May 2007 04:47:09 -0500 } Received: (from mail-at-localhost) } by ns.microscopy.com (8.12.11.20060308/8.12.11/Submit) id l4M9l9uH018525; } Tue, 22 May 2007 04:47:09 -0500 } Date: Tue, 22 May 2007 04:47:09 -0500 } Message-Id: {200705220947.l4M9l9uH018525-at-ns.microscopy.com} } To: jquinn-at-www.matscieng.sunysb.edu } From: nizets2-at-yahoo.com } Reply-to: nizets2-at-yahoo.com } X-Resent-From: "Microscopy Listserver" {microscopy-at-microscopy.com} } Subject: [Microscopy] basic SEM: imaging salt crystals } Errors-To: MicroscopyListSpamFilter-at-microscopy.com } X-lewp: MicroscopyListSpam NAGS } Status: R } } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Dear listers, } } Here are some basic questions from the basic SEM user } I am: I would like to visualize salt crystals by SEM } and make EDX analysis if possible. I just leave a drop } of salt solution on a carbon tab dry, that's all! } My questions: } } - Do I need to carbon-coat the crystals? } - Is there a risk to evaporate the crystals under the } electron beam and make the column dirty? (I usually } don't use HV higher than 15 kV). Would carbon coating } help dissipate the heat in this regard? } } Now another question related to SEM but not to } crystals: } When I analyse fine particles by EDX, I never know if } I analyse only the particle or also part of the } material which is behind the particle. Is there a way } to determine the depth of analysis of the material } studied? I suppose that it depends on the HV and on } the nature of the material itself, but I already would } be happy to have a order of magnitude. Is this 10 nm } or 2 µm deep? } Lets say I analyze at 15kV (1) NaCl crystals (2) } Quartz (3) Feldpar (4) Copper, what would be the } expected "depth of analysis" in these samples? } Does it depend on the size of the beam? } } Best regards, } } Stephane } } } } } } } } } ____________________________________________________________________________________ } Need Mail bonding? } Go to the Yahoo! Mail Q&A for great tips from Yahoo! Answers users. } http://answers.yahoo.com/dir/?link=list&sid=396546091 } } ==============================Original Headers============================== } 13, 19 -- From nizets2-at-yahoo.com Tue May 22 04:46:12 2007 } 13, 19 -- Received: from web37405.mail.mud.yahoo.com (web37405.mail.mud.yahoo.com [209.191.91.137]) } 13, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l4M9kCVc017352 } 13, 19 -- for {microscopy-at-microscopy.com} ; Tue, 22 May 2007 04:46:12 -0500 } 13, 19 -- Received: (qmail 54717 invoked by uid 60001); 22 May 2007 09:46:08 -0000 } 13, 19 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; } 13, 19 -- s=s1024; d=yahoo.com; } 13, 19 -- h=X-YMail-OSG:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding:Message-ID; } 13, 19 -- b=zBbqWoaMZ9PlXpKnS7/04OoVZZIK8DFKFHcyjE3Fb/m3Zl4auB81Rs10Qpyy2eM+Nm+m0B7lAIn+OTWrzWZNMScRq4hkKwM1at2lB+Ok+wl3FvDKesHuEIr4DQMXBSph2qIICP+SmycYwMy8nLABeHvgi3DVBD88TCbeU3lTI7w=; } 13, 19 -- X-YMail-OSG: _Sy6fjQVM1n_aA9U2sOeO5UGkSLDD5RhnQ74_ILWjiw02eS6PDlVt_Y7dn7dCMkDiw-- } 13, 19 -- Received: from [80.122.101.102] by web37405.mail.mud.yahoo.com via HTTP; Tue, 22 May 2007 02:46:08 PDT } 13, 19 -- Date: Tue, 22 May 2007 02:46:08 -0700 (PDT) } 13, 19 -- From: Stephane Nizet {nizets2-at-yahoo.com} } 13, 19 -- Subject: basic SEM: imaging salt crystals } 13, 19 -- To: microscopy-at-microscopy.com } 13, 19 -- MIME-Version: 1.0 } 13, 19 -- Content-Type: text/plain; charset=iso-8859-1 } 13, 19 -- Content-Transfer-Encoding: 8bit } 13, 19 -- Message-ID: {210050.53719.qm-at-web37405.mail.mud.yahoo.com} } ==============================End of - Headers============================== }
} From mail-at-ns.microscopy.com Tue May 22 05:45:52 2007 } Date: Tue, 22 May 2007 04:47:09 -0500 } To: jquinn-at-www.matscieng.sunysb.edu } From: nizets2-at-yahoo.com } Reply-to: nizets2-at-yahoo.com } X-Resent-From: "Microscopy Listserver" {microscopy-at-microscopy.com} } Subject: [Microscopy] basic SEM: imaging salt crystals } Errors-To: MicroscopyListSpamFilter-at-microscopy.com } X-lewp: MicroscopyListSpam NAGS } } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Dear listers, } } Here are some basic questions from the basic SEM user } I am: I would like to visualize salt crystals by SEM } and make EDX analysis if possible. I just leave a drop } of salt solution on a carbon tab dry, that's all! } My questions: } } - Do I need to carbon-coat the crystals? } - Is there a risk to evaporate the crystals under the } electron beam and make the column dirty? (I usually } don't use HV higher than 15 kV). Would carbon coating } help dissipate the heat in this regard? } } Now another question related to SEM but not to } crystals: } When I analyse fine particles by EDX, I never know if } I analyse only the particle or also part of the } material which is behind the particle. Is there a way } to determine the depth of analysis of the material } studied? I suppose that it depends on the HV and on } the nature of the material itself, but I already would } be happy to have a order of magnitude. Is this 10 nm } or 2 µm deep? } Lets say I analyze at 15kV (1) NaCl crystals (2) } Quartz (3) Feldpar (4) Copper, what would be the } expected "depth of analysis" in these samples? } Does it depend on the size of the beam? } } Best regards, } } Stephane } } } } } } } } } ____________________________________________________________________________________ } Need Mail bonding? } Go to the Yahoo! Mail Q&A for great tips from Yahoo! Answers users. } http://answers.yahoo.com/dir/?link=list&sid=396546091 } } ==============================Original Headers============================== } 13, 19 -- From nizets2-at-yahoo.com Tue May 22 04:46:12 2007 } 13, 19 -- Received: from web37405.mail.mud.yahoo.com (web37405.mail.mud.yahoo.com [209.191.91.137]) } 13, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l4M9kCVc017352 } 13, 19 -- for {microscopy-at-microscopy.com} ; Tue, 22 May 2007 04:46:12 -0500 } 13, 19 -- Received: (qmail 54717 invoked by uid 60001); 22 May 2007 09:46:08 -0000 } 13, 19 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; } 13, 19 -- s=s1024; d=yahoo.com; } 13, 19 -- h=X-YMail-OSG:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding:Message-ID; } 13, 19 -- b=zBbqWoaMZ9PlXpKnS7/04OoVZZIK8DFKFHcyjE3Fb/m3Zl4auB81Rs10Qpyy2eM+Nm+m0B7lAIn+OTWrzWZNMScRq4hkKwM1at2lB+Ok+wl3FvDKesHuEIr4DQMXBSph2qIICP+SmycYwMy8nLABeHvgi3DVBD88TCbeU3lTI7w=; } 13, 19 -- X-YMail-OSG: _Sy6fjQVM1n_aA9U2sOeO5UGkSLDD5RhnQ74_ILWjiw02eS6PDlVt_Y7dn7dCMkDiw-- } 13, 19 -- Received: from [80.122.101.102] by web37405.mail.mud.yahoo.com via HTTP; Tue, 22 May 2007 02:46:08 PDT } 13, 19 -- Date: Tue, 22 May 2007 02:46:08 -0700 (PDT) } 13, 19 -- From: Stephane Nizet {nizets2-at-yahoo.com} } 13, 19 -- Subject: basic SEM: imaging salt crystals } 13, 19 -- To: microscopy-at-microscopy.com } 13, 19 -- MIME-Version: 1.0 } 13, 19 -- Content-Type: text/plain; charset=iso-8859-1 } 13, 19 -- Content-Transfer-Encoding: 8bit } 13, 19 -- Message-ID: {210050.53719.qm-at-web37405.mail.mud.yahoo.com} } ==============================End of - Headers============================== }
==============================Original Headers============================== 13, 12 -- From jquinn-at-www.matscieng.sunysb.edu Tue May 22 09:30:23 2007 13, 12 -- Received: from www.matscieng.sunysb.edu (www.matscieng.sunysb.edu [129.49.36.33]) 13, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4MEUM0l004297 13, 12 -- for {microscopy-at-microscopy.com} ; Tue, 22 May 2007 09:30:22 -0500 13, 12 -- Received: (from jquinn-at-localhost) 13, 12 -- by www.matscieng.sunysb.edu (8.11.6/8.11.6) id l4MESwq17328 13, 12 -- for microscopy-at-microscopy.com; Tue, 22 May 2007 10:28:58 -0400 13, 12 -- Date: Tue, 22 May 2007 10:28:58 -0400 13, 12 -- From: Jim Quinn {jquinn-at-www.matscieng.sunysb.edu} 13, 12 -- Message-Id: {200705221428.l4MESwq17328-at-www.matscieng.sunysb.edu} 13, 12 -- To: microscopy-at-microscopy.com 13, 12 -- Subject: re: SEM and EDAX ==============================End of - Headers==============================
cgarber-at-2spi.com wrote: } {snip} } } I also agree with Roger that idle "chit-chat" can be hazardous when one is trying to do work with potentially dangerous tools at the same time. } } Disclaimer: SPI Supplies distributes the GEM single edge "scientific" razor blades so we would have a vested interestin seeing more of our customers using this type of blade. } } Chuck } } =================================================== } Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 } President } SPI SUPPLIES FAX: 1-(610)-436-5755 } PO BOX 656 e-mail: cgarber-at-2spi.com } West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com } } WWW: http://www.2spi.com } You are the key to your own safety. Not the safety officer, not the rules in the lab, not the no slip floors, no spill containers, hoods or anything else in the end your safety rests in on you. Nothing can make you safe from you if your not safe in the way you do things. Just take the case of Lucky. He a man that has cut off the fingers of one hand, ruined the joints in the other arm , his back and more. He managed to blow up two guns reloading ammunition and runs his boat on the rocks nearly every time he goes out. Of all the fishing rods I have broken Lucky stepped on all 3. Nothing can be made safe for Lucky to use. I call him Lucky because he turned his head just a bolt snapped off a machine he was abusing and knocked out all his front teeth. Had he not turned around to look at the machine the bolt would have hit the base of his scull and killed him. I don't get close to Lucky when he is working or hunting and we fish in my boat.
When cutting proper tools for the job make it safer and easier. I have too many tools. I am obsessive about tools being sharp and properly shaped for the job. But I don't have many scars on my hands from knives. I do use double edged razor blade cut into 4 pieces an mounted in a handle for some things as described by Anna Teetsov at McCrone Associates in "Quarter-Razor Blade for Hand Sectioning" in the Prep School Section of Modern Microscopy www.modernmicroscopy.com/main.asp?article=50 {http://www.modernmicroscopy.com/main.asp?article=50} . I sure wouldn't use them with out a handle. I also find Schick razor blades useful in a handle thought somewhat expensive and hard to find. I sure wouldn't use either one with out a good handle on it.
In 30 years of working at hazardous work, farming, machine shops, welding and such before coming into a slightly safer lab environment every injury I was involved with was from not paying attention to what I was doing. Trying to hurry was the worst offender. Safety is in the person doing the work. Taking my time and planing ahead have got me though life with all my fingers and the only broken bone is my nose when I came out of the bath room and tripped over the kids dog we were keeping for them while they went to China 2 years ago.
The only serious accident I have been involved in was when I was taking a neighbor with brain damaged to the brain stem to a meeting and he fell and broke his pelvis as he got out of the car. It was years after I retired and we were talking and I failed to stand behind him as I normally do with anyone that has trouble with getting in and out of a car so I can catch them should they fall. I use a cane when I have that much trouble getting around and that's 2 or 3 times a year but many people let pride get in the way of good sense on things like that. Pride has no place where safety is concerned. Anything that make you less safe needs to be dealt with once an for all. If it is miss placed pride, contempt for a naive safety officers, cultural custom, urgency or anything that distracts you from the the task at hand you need to deal with it now.
No one can make you safe but you.
Gordon Gordon Couger Microscope documentation Archive http://science-info.net/
==============================Original Headers============================== 8, 19 -- From gcouger-at-science-info.net Tue May 22 11:40:16 2007 8, 19 -- Received: from smtp106.biz.mail.mud.yahoo.com (smtp106.biz.mail.mud.yahoo.com [68.142.200.254]) 8, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l4MGeGbI019072 8, 19 -- for {Microscopy-at-microscopy.com} ; Tue, 22 May 2007 11:40:16 -0500 8, 19 -- Received: (qmail 97146 invoked from network); 22 May 2007 16:40:16 -0000 8, 19 -- Received: from unknown (HELO ?192.168.1.100?) (gcouger-at-science-info.net-at-74.195.225.38 with plain) 8, 19 -- by smtp106.biz.mail.mud.yahoo.com with SMTP; 22 May 2007 16:40:15 -0000 8, 19 -- X-YMail-OSG: 2xcU98cVM1l.KknqsYvD0TsxXvS2lE1_8cd.pib7x028ripj7beRfkGdc5RgZnxt2cSeOSz7tW387AsrqFoFVLVQLtGhb.8BjExvYe57oqLGsqRmomw- 8, 19 -- Message-ID: {46531D10.5060605-at-science-info.net} 8, 19 -- Date: Tue, 22 May 2007 11:40:48 -0500 8, 19 -- From: Gordon Couger {gcouger-at-science-info.net} 8, 19 -- User-Agent: Thunderbird 1.5.0.10 (Macintosh/20070221) 8, 19 -- MIME-Version: 1.0 8, 19 -- To: cgarber-at-2spi.com, Microscopy {Microscopy-at-microscopy.com} 8, 19 -- Subject: Re: [Microscopy] Cutting tools and hazards 8, 19 -- References: {200705220727.l4M7RBHC013434-at-ns.microscopy.com} 8, 19 -- In-Reply-To: {200705220727.l4M7RBHC013434-at-ns.microscopy.com} 8, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 8, 19 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
On May 21, 2007, at 2:47 PM, c.r.vandenbrom-at-bham.ac.uk wrote:
} 1. What materials are typically used to make holey or lacey films. } Just amorphous carbon and formvar? } } 2. What are the typical dimensions attainable for holey or lacey } films, in terms of film thickness and hole size. For our purposes, } 20-40 nm hole diameter would be ideal, but 100 - 200 nm would be very } helpful already. } } 3. Are there good descriptions of the preparation or availability of } such holey films? } } 4. Does anyone have experience in supporting lipid bilayers in } different ways, e.g. for TEM investigation? } Dear Coen, 1. Typically holey formvar is produced, carbon is evaporated onto it, and then the formvar is dissolved away, leaving a holey carbon support film. This is unsuitable for supporting lipid bilayers for at least two reasons; first, the carbon will not be hydrophobic enough that lipid would wet it, and second, the areas of the holes are not well-defined enough to measure ion current as a function of bilayer area accurately. I know of techniques that can change the nature of the carbon--glow-discharging to increase hydrophyllicity, and letting the carbon age to increase hydrophobicity--but I do not think that the carbon ever gets truly hydrophobic. Once one has a holey formvar film, one can either leave it uncoated or evaporate or otherwise coat it with a variety of substances other than carbon, so perhaps a suitable hydrophobic coating can be found. 2. One can vary the hole size during preparation. Generally, one suspends hydrophyllic droplets in the formvar, so that when the formvar dries, the droplets leave holes where the formvar has been excluded. This results in holes of about the size of the droplets, which are typically of um dimension, not nm. However, holes can be produced by irradiation, so perhaps holes in your desired size range are possible. 3. Yes, but I don't have one at hand. 4. I have just dissolved the lipid in a suitable solvent, put a drop across a hole in a teflon wafer, let the dissolved lipid spread until a bilayer formed across the hole, and lowered the wafer into the water so that the wafer served to separate the two sides of a compartment or added water to both sides of the compartment after the wafer was in place. This way the area of the bilayer is known, and one could use a wafer with many holes of the desired total area. There are likely to be companies that can construct plastic wafers with appropriately sized holes, and maybe one or more of them will contact you through the list. Good luck. Yours, Bill Tivol, PhD EM Scientist Electron Cryo-Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
==============================Original Headers============================== 4, 22 -- From tivol-at-caltech.edu Tue May 22 12:03:13 2007 4, 22 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4MH3Dsj031003 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 22 May 2007 12:03:13 -0500 4, 22 -- Received: from water-dog.its.caltech.edu (water-dog [192.168.1.26]) 4, 22 -- by earth-ox-postvirus (Postfix) with ESMTP id 9CF3F1A9C7; 4, 22 -- Tue, 22 May 2007 10:03:05 -0700 (PDT) 4, 22 -- Received: from [192.168.159.233] (jpix-01.caltech.edu [131.215.2.133]) 4, 22 -- by fire-ox.its.caltech.edu (Postfix) with ESMTP id 8743E12D45; 4, 22 -- Tue, 22 May 2007 10:03:03 -0700 (PDT) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 22 -- In-Reply-To: {200705212147.l4LLl6Qs027444-at-ns.microscopy.com} 4, 22 -- References: {200705212147.l4LLl6Qs027444-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {b5d195f9838417b78e3c1ed77be94e69-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] Supporting lipid bilayers with holey carbon? 4, 22 -- Date: Tue, 22 May 2007 10:15:03 -0700 4, 22 -- To: microscopy-at-msa.microscopy.com, c.r.vandenbrom-at-bham.ac.uk 4, 22 -- X-Mailer: Apple Mail (2.624) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on water-dog by amavisd-2.4.5 ==============================End of - Headers==============================
On May 22, 2007, at 2:46 AM, nizets2-at-yahoo.com wrote:
} Here are some basic questions from the basic SEM user } I am: I would like to visualize salt crystals by SEM } and make EDX analysis if possible. I just leave a drop } of salt solution on a carbon tab dry, that's all! } My questions: } } - Do I need to carbon-coat the crystals? } - Is there a risk to evaporate the crystals under the } electron beam and make the column dirty? (I usually } don't use HV higher than 15 kV). Would carbon coating } help dissipate the heat in this regard? } } Now another question related to SEM but not to } crystals: } When I analyse fine particles by EDX, I never know if } I analyse only the particle or also part of the } material which is behind the particle. Is there a way } to determine the depth of analysis of the material } studied? I suppose that it depends on the HV and on } the nature of the material itself, but I already would } be happy to have a order of magnitude. Is this 10 nm } or 2 µm deep? } Lets say I analyze at 15kV (1) NaCl crystals (2) } Quartz (3) Feldpar (4) Copper, what would be the } expected "depth of analysis" in these samples? } Does it depend on the size of the beam? } Dear Stephane, If the crystals are conductive, you do not need to coat them, and if they are not, you could examine them at a voltage for which the production and escape of secondary electrons is the same as the beam electrons deposited in the crystal, giving no net change in charge. This might not be sufficient voltage to generate particular X-ray lines, however. Heat conductivity is generally high in substances for which charge conductivity is high, and for substances with low heat conductivity, using a low beam current with longer times of analysis can ameliorate specimen heating, so I would advise against carbon coating. In any case, carbon coating will not help with heat generated too far below the surface of the sample, but this may not be an issue. David Joy has written programs to determine the analysis volumes for various substances using electrons of various energies, and I think that these are freely available. For 15 kV or below, the depth would be closer to 10 nm than to 2 um--I don't have access to my tables of electron stopping powers, so I can't give you a better answer. The depth does not depend strongly on the size of the beam; each electron acts independently. If, however, excess charge is not dissipated, and the extent of the charged volume is larger and the charge per unit volume is smaller for a wider, but less intense beam, there may be a difference in the voltage at the surface of the specimen that could lead to changes in the depth of analysis. In the absence of charging effects--which will obviously be different for quartz than for copper--the penetration depth of electrons is approximately the same when measured in units of mg/cm^2 for all materials. It is greater in higher-Z materials, since they have a higher neutron/proton ratio, which adds mass but not stopping power. Yours, Bill Tivol, PhD EM Scientist Electron Cryo-Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
==============================Original Headers============================== 5, 23 -- From tivol-at-caltech.edu Tue May 22 12:31:53 2007 5, 23 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 5, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4MHVrAb010561 5, 23 -- for {microscopy-at-msa.microscopy.com} ; Tue, 22 May 2007 12:31:53 -0500 5, 23 -- Received: from water-dog.its.caltech.edu (water-dog [192.168.1.26]) 5, 23 -- by earth-ox-postvirus (Postfix) with ESMTP id 1F32F1AB2D 5, 23 -- for {microscopy-at-msa.microscopy.com} ; Tue, 22 May 2007 10:31:52 -0700 (PDT) 5, 23 -- Received: from [192.168.159.233] (jpix-01.caltech.edu [131.215.2.133]) 5, 23 -- by earth-ox.its.caltech.edu (Postfix) with ESMTP id 276F81AB2B 5, 23 -- for {microscopy-at-msa.microscopy.com} ; Tue, 22 May 2007 10:31:51 -0700 (PDT) 5, 23 -- Mime-Version: 1.0 (Apple Message framework v624) 5, 23 -- In-Reply-To: {200705220946.l4M9kK2w017493-at-ns.microscopy.com} 5, 23 -- References: {200705220946.l4M9kK2w017493-at-ns.microscopy.com} 5, 23 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 5, 23 -- Message-Id: {4d95550528f749a41a406af0272ee28f-at-caltech.edu} 5, 23 -- From: Bill Tivol {tivol-at-caltech.edu} 5, 23 -- Subject: Re: [Microscopy] basic SEM: imaging salt crystals 5, 23 -- Date: Tue, 22 May 2007 10:43:51 -0700 5, 23 -- To: microscopy-at-msa.microscopy.com 5, 23 -- X-Mailer: Apple Mail (2.624) 5, 23 -- X-Spam-Scanned: at Caltech-ITS on water-dog by amavisd-2.4.5 5, 23 -- Content-Transfer-Encoding: 8bit 5, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4MHVrAb010561 ==============================End of - Headers==============================
-- Coen: Try this as a technique for making holey carbon films: http://cryoem.ucsd.edu/procedures/H-carbonfilm.shtm
I wouldn't call it the final and absolute way to do it but it is the one we teach to our students and it works well. The holes we get are of varying size from less than a micron in diameter on up to several microns. If you want the smallest holes possible I would suggest sonnicating for a much longer time than suggested in the procedure.
Norm
______________________________________________________________ Norm Olson Cryoelectron Microscopy Facilities Manager 1510 Bonner Hall Department of Chemistry & Biochemistry, MC-0378 University of California San Diego La Jolla, CA 92093-0378 nholson-at-ucsd.edu http://cryoem.ucsd.edu Cell: (858)220-2183 (858)822-6718 - Office; (858)534-5846 - Fax ______________________________________________________________
3. Are there good descriptions of the preparation or availability of such holey films?
Thank you all for your help!
With kind regards,
Coen van den Brom
----------------------------
dr CR van den Brom Department of Nanoscale Chemistry School of Chemistry University of Birmingham Edgbaston Birmingham B15 2TT United Kingdom
Why a salt solution? You can just drop salt crystals onto a sticky tab on a stub and image a bunch of crystals. Different salts look different. Table salt is like a face centered cubic while marine salt is layered as is Kosher salt.
For EDS, the Cl shells are the ones to get set up for. Na Ka is at 1.041KeV while Cl Ka is at 2.621KeV. The only other transitions are K beta. Thus, about 6-8KV should be enough KV to get all peaks and do quant. To get just the/a crystal, do spot mode and put the spot on a crystal.
I would coat the crystals with Au/Pd or Pt or Pd. Au is probably best since its La peak is at 9.710KeV and its Ma 2.120KeV which puts that peak in between Na Ka and Cl Ka. I would not coat with C since if your beam is off of the crystal, you would not necessarily know if the C is from the coating or the sticky tab. It probably would be a giveaway since the tab C peak would be higher (much) than the coating C peak. But just eliminate this issue by not using C.
With coating and relatively low KV, you can get nice surface morphology. Counts will probably be low but if beam position is stable, just collect for enough time for good results.
The Electron Flight Simulator will tell you how deep a particular KV beam will penetrate into a specified material.
gary g.
At 01:47 AM 5/22/2007, you wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 10, 22 -- From gary-at-gaugler.com Tue May 22 13:26:37 2007 10, 22 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 10, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l4MIQaMg002370 10, 22 -- for {microscopy-at-microscopy.com} ; Tue, 22 May 2007 13:26:37 -0500 10, 22 -- Message-Id: {200705221826.l4MIQaMg002370-at-ns.microscopy.com} 10, 22 -- Received: (qmail 4107 invoked from network); 22 May 2007 11:26:36 -0700 10, 22 -- Received: by simscan 1.1.0 ppid: 4100, pid: 4102, t: 0.1374s 10, 22 -- scanners: regex: 1.1.0 attach: 1.1.0 10, 22 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 10, 22 -- by qsmtp3 with SMTP; 22 May 2007 11:26:36 -0700 10, 22 -- X-Mailer: QUALCOMM Windows Eudora Version 7.1.0.9 10, 22 -- Date: Tue, 22 May 2007 11:26:30 -0800 10, 22 -- To: nizets2-at-yahoo.com 10, 22 -- From: Gary Gaugler {gary-at-gaugler.com} 10, 22 -- Subject: Re: [Microscopy] basic SEM: imaging salt crystals 10, 22 -- Cc: MSA listserver {microscopy-at-microscopy.com} 10, 22 -- In-Reply-To: {200705220947.l4M9lxML020213-at-ns.microscopy.com} 10, 22 -- References: {200705220947.l4M9lxML020213-at-ns.microscopy.com} 10, 22 -- Mime-Version: 1.0 10, 22 -- Content-Type: text/plain; charset=iso-8859-1; format=flowed; x-avg-checked=avg-ok-7F1E14CA 10, 22 -- Content-Transfer-Encoding: 8bit 10, 22 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4MIQaMg002370 ==============================End of - Headers==============================
I have already seen a number of good answers, but I will chime in anyway.
I would suggest coating the samples unless you are operating in variable pressure mode (which may not be an option for you). I recall that NaCl charges as well as most any insulator. I don't imagine you would be doing quant on the sample since the lack of a known geometry would lead to substantial uncertainty in the results. Gary made a good point that it would be nice to use an element for coating that is only present in the coating. Otherwise, there will be uncertainty in the source of the peaks. I would expect some difference in the relative peak intensities depending on which element is used.
I wouldn't worry much about vaporizing the salt. (Maybe I should.) I don't expect you will be using nanoamps of current. Once again, VP mode would help minimize or prevent contamination.
I don't have the latest and greatest Monte Carlo programs. I do have an old one derived from David Joy's work. I ran a simulation on S at 15 kV and I estimate the excitation depth to be slightly less than 1 um. Running the same simulation for Na, I found the depth to be more like 2 um. For Cu, I found the depth to be on the order of 0.3 um. Therefore, I generally quote penetration depths of hundreds of nm up to a couple of microns. The dimensions of the interaction volume dwarf the spot size so much that I don't often bother considering it in calculating interaction volume.
As a fun little exercise, you might lay down a gold layer on copper or some other metal. The interaction volume for gold appears to be slightly less than 0.2 um (200 nm). Since we routinely coat samples with 10 nm of gold, we need a deeper penetration depth if we are ever going to see the sample through the gold. We don't have a problem seeing the sample unless we are using a very thick layer of gold or a low voltage. A 3 kV beam would have difficulty penetrating 10 nm of gold, but there would not be much excitation of sample x-rays at that low a voltage anyway.
Warren
-----Original Message----- X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] Sent: Tuesday, May 22, 2007 4:47 AM To: wesaia-at-iastate.edu
This Question was submitted to Ask-A-Microscopist by (hopechen-at-u.washington.edu) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, May 22, 2007 at 18:17:42 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both hopechen-at-u.washington.edu as well as to the Microscopy Listserver ---------------------------------------------------------------------------
I try to set up a high-speed microphotography system using an ultra-high speed camera and an inverted microscope. I'm now facing a problem that is how to incorporate flash tube light source with the microscope. The light source we has is Photogenic Radio Powerlight 2500DRUV and the flash tube is Photogenic C4-19 Flashtube. The flash tube is annular with an outside diameter about 1.8 inch and inner diameter about 1 inch. Our requirements to the microscope illumination system are: 1) collect as much light as possible to increase the illumination light intensity, which hopefully could allow us to adjust our high-speed camera's exposure time to as small as 10 ns, and 2) help us to get high resolution images. Iím thinking of using Kohler illumination, but due to the light source is a large annular light source I couldnít get idea Kohler illumination. Nelsonian illumination focus light to the specimen, so this illumination method might be better than Kohler illumination in terms of light intensity, but then I need to focus an annular light to a point. May I ask for your suggestions?
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If no one come up with a better idea, you may consider trying a truncated cone of Plexiglas that has one end large enough to capture most of the light from the flash tube and the other the size you need for the light source of the scope and fitted in the proper place over the condenser may work. You may have to frost the small end of the cone or cut the large end at an angle of the central axis of the microscope lighting to get the light as even as you want.
The cone works on the principle of Total Internal Reflection. I would use Plexiglas because it can be fire polished by finishing it with 400 grit wet/dry silicon carbide sandpaper and then a flame will melt the surface to a high polish. It takes a few tires to get the fire polishing down but you can clean up the mess and do it over a lot of times. Just keep the flame moving and the part spinning. The fire polishing happens very fast at about 180-200 C.
Using plastic any machine shop or anyone with a lathe can do the job in a few minutes and you don't need a optical shop to do the work.
Plexiglas is much more transparent to light than glass and doesn't block Ultra Violet. So it works with florescent light as well.
Good luck Gordon Gordon Couger Microscope documentation Archive http://science-info.net/
hopechen-at-u.washington.edu wrote: } } Email: hopechen-at-u.washington.edu } Name: Hope Chen } } Organization: University of Washington } } Education: Graduate College } } Location: Seattle. WA US } } Title: microscope illumination } } Question: Dear Microscopist, } } I try to set up a high-speed microphotography system using an ultra-high speed camera and an inverted microscope. I'm now facing a problem that is how to incorporate flash tube light source with the microscope. The light source we has is Photogenic Radio Powerlight 2500DRUV and the flash tube is Photogenic C4-19 Flashtube. The flash tube is annular with an outside diameter about 1.8 inch and inner diameter about 1 inch. Our requirements to the microscope illumination system are: 1) collect as much light as possible to increase the illumination light intensity, which hopefully could allow us to adjust our high-speed camera's exposure time to as small as 10 ns, and 2) help us to get high resolution images. } Iím thinking of using Kohler illumination, but due to the light source is a large annular light source I couldnít get idea Kohler illumination. Nelsonian illumination focus light to the specimen, so this illumination method might be better than Kohler illumination in terms of light intensity, but then I need to focus an annular light to a point. } May I ask for your suggestions? } } send with thanks, } Hope } } }
==============================Original Headers============================== 12, 20 -- From gcouger-at-science-info.net Wed May 23 01:23:33 2007 12, 20 -- Received: from smtp104.biz.mail.mud.yahoo.com (smtp104.biz.mail.mud.yahoo.com [68.142.200.252]) 12, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l4N6NWl2028178 12, 20 -- for {microscopy-at-microscopy.com} ; Wed, 23 May 2007 01:23:33 -0500 12, 20 -- Received: (qmail 83156 invoked from network); 23 May 2007 06:23:32 -0000 12, 20 -- Received: from unknown (HELO ?192.168.1.100?) (gcouger-at-science-info.net-at-74.195.225.38 with plain) 12, 20 -- by smtp104.biz.mail.mud.yahoo.com with SMTP; 23 May 2007 06:23:32 -0000 12, 20 -- X-YMail-OSG: DfdrI4wVM1kkamvqiTLJXpuwwB6qJaMiSd7oGxwxVsx.Ffkx3qVFLiwcb.hg.SGGtbfbMczx_IoS9bRmlqdUitiWxe912w0yo9iTGMrdsGIthd3x330- 12, 20 -- Message-ID: {4653DE05.4060902-at-science-info.net} 12, 20 -- Date: Wed, 23 May 2007 01:24:05 -0500 12, 20 -- From: Gordon Couger {gcouger-at-science-info.net} 12, 20 -- User-Agent: Thunderbird 1.5.0.10 (Macintosh/20070221) 12, 20 -- MIME-Version: 1.0 12, 20 -- To: hopechen-at-u.washington.edu, microscopy-at-microscopy.com 12, 20 -- Subject: Re: [Microscopy] AskAMicroscopist:High Speed microscope illumination 12, 20 -- References: {200705230240.l4N2emt9023509-at-ns.microscopy.com} 12, 20 -- In-Reply-To: {200705230240.l4N2emt9023509-at-ns.microscopy.com} 12, 20 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 12, 20 -- Content-Transfer-Encoding: 8bit 12, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4N6NWl2028178 ==============================End of - Headers==============================
I have a faculty member who wants to do a perfusion fix of brain (it's a rat or mouse I forget which). Normally I avoid using cacodylate buffer because of the arsenic. However, in the dim recesses of my memory I seem to recall that phosphate buffer is very prone to precipitate formation in brain tissue. Now I don't know if I'm remembering this correctly. So, what are your opinions? Would it be better to use a cacodylate buffer or should I stick with the safer less toxic phosphate buffer? As always thanks in advance for your help.
Tom Bargar University of Nebraska Medical Center Core Electron Microscopy Research Facility 986395 Nebraska Medical Center Omaha, NE 68198-6395 402-559-7347 tbargar-at-unmc.edu
==============================Original Headers============================== 5, 20 -- From tbargar-at-unmc.edu Wed May 23 08:44:23 2007 5, 20 -- Received: from zixvpm02.unmc.edu (zixvpm02.unmc.edu [192.198.54.127]) 5, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4NDiNN3016054 5, 20 -- for {Microscopy-at-MSA.Microscopy.com} ; Wed, 23 May 2007 08:44:23 -0500 5, 20 -- Received: from zixvpm02.unmc.edu (ZixVPM [127.0.0.1]) 5, 20 -- by Outbound.unmc.edu (Proprietary) with ESMTP id EDE07A0063 5, 20 -- for {Microscopy-at-MSA.Microscopy.com} ; Wed, 23 May 2007 08:44:22 -0500 (CDT) 5, 20 -- Received: from unmcnotes01.unmc.edu (host-137-197-103-35.unmc.edu [137.197.103.35]) 5, 20 -- by zixvpm02.unmc.edu (Proprietary) with ESMTP id C206139804A 5, 20 -- for {Microscopy-at-MSA.Microscopy.com} ; Wed, 23 May 2007 08:44:21 -0500 (CDT) 5, 20 -- Subject: Cacodylate buffer vs. phosphate buffer 5, 20 -- To: Microscopy-at-MSA.Microscopy.com 5, 20 -- X-Mailer: Lotus Notes Release 7.0.1 January 17, 2006 5, 20 -- Message-ID: {OFDC28CBBD.26B17965-ON862572E4.004ACE85-862572E4.004B786B-at-unmc.edu} 5, 20 -- From: Tom W Bargar {tbargar-at-unmc.edu} 5, 20 -- Date: Wed, 23 May 2007 08:44:20 -0500 5, 20 -- X-MIMETrack: Serialize by Router on UNMCNOTES01.UNMC.EDU/Servers/UNEBR at 05/23/2007 08:44:21 5, 20 -- AM 5, 20 -- MIME-Version: 1.0 5, 20 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
I have never used it for perfusion fixation but HEPES buffer works well in my standard fixes with 2% paraformaldehyde or 2% paraformaldehyde + 2.5% glutaraldehyde. Alternatively, PIPES is used by other researchers with success. I see no argument for retaining use of cacodylate. Good luck, Tom
At 08:46 AM 05/23/07, you wrote:
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For EDS analysis, if you coat your crystals with a high Z material, and your analysis software dont' have an "sample coating" option, you'll probably will have wrong results on crystals like NaCl. It's always difficult to have accurate results on Na (it evaoprate under the beam, but not enough to polluate the SEM !), but with gold coating, absorption from the Na-k by the gold layer will be much stronger than that from Cl-k, and the balance Na-Cl will be worse. I think that carbon is the less problematic coating for such analysis, and I would not use Au or Pt. Of coarse, imaging can be less nice than with Au, Pt or Ir coating.
For Monte Carlo simulation, you can download Casino (http://www.gel.usherbrooke.ca/casino/), with has some nice possibility, like multiple energies simulation. I use it complementary to Electron Flight Simulator.
A simple test in particule (or thin layer) analysis, is to put the particules on a substrate which don't have any element from the particules (of coarse, you must have some idees about these particules, and yes, you don't have any, that's what your are looking for !) and to perform measurement with different energies, from say 5 to 20 keV. As long as you see the x-ray lines of the subtrate in the spectrum, the primary energie is too hight. By the way, it is suffisent that one element from the substrate does not exist in the particule, if possible with a line at low energie, to be able to see it in the spectrum at all energies (Cu-L par exemple).
Hope it helps
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
wesaia-at-iastate.edu a écrit : } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } I have already seen a number of good answers, but I will chime in anyway. } } I would suggest coating the samples unless you are operating in variable pressure mode (which may not be an option for you). I recall that NaCl charges as well as most any insulator. I don't imagine you would be doing quant on the sample since the lack of a known geometry would lead to substantial uncertainty in the results. Gary made a good point that it would be nice to use an element for coating that is only present in the coating. Otherwise, there will be uncertainty in the source of the peaks. I would expect some difference in the relative peak intensities depending on which element is used. } } I wouldn't worry much about vaporizing the salt. (Maybe I should.) I don't expect you will be using nanoamps of current. Once again, VP mode would help minimize or prevent contamination. } } I don't have the latest and greatest Monte Carlo programs. I do have an old one derived from David Joy's work. I ran a simulation on S at 15 kV and I estimate the excitation depth to be slightly less than 1 um. Running the same simulation for Na, I found the depth to be more like 2 um. For Cu, I found the depth to be on the order of 0.3 um. Therefore, I generally quote penetration depths of hundreds of nm up to a couple of microns. The dimensions of the interaction volume dwarf the spot size so much that I don't often bother considering it in calculating interaction volume. } } As a fun little exercise, you might lay down a gold layer on copper or some other metal. The interaction volume for gold appears to be slightly less than 0.2 um (200 nm). Since we routinely coat samples with 10 nm of gold, we need a deeper penetration depth if we are ever going to see the sample through the gold. We don't have a problem seeing the sample unless we are using a very thick layer of gold or a low voltage. A 3 kV beam would have difficulty penetrating 10 nm of gold, but there would not be much excitation of sample x-rays at that low a voltage anyway. } } Warren } } -----Original Message----- } X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] } Sent: Tuesday, May 22, 2007 4:47 AM } To: wesaia-at-iastate.edu } Subject: [Microscopy] basic SEM: imaging salt crystals } } Dear listers, } } Here are some basic questions from the basic SEM user } I am: I would like to visualize salt crystals by SEM } and make EDX analysis if possible. I just leave a drop } of salt solution on a carbon tab dry, that's all! } My questions: } } - Do I need to carbon-coat the crystals? } - Is there a risk to evaporate the crystals under the } electron beam and make the column dirty? (I usually } don't use HV higher than 15 kV). Would carbon coating } help dissipate the heat in this regard? } } Now another question related to SEM but not to } crystals: } When I analyse fine particles by EDX, I never know if } I analyse only the particle or also part of the } material which is behind the particle. Is there a way } to determine the depth of analysis of the material } studied? I suppose that it depends on the HV and on } the nature of the material itself, but I already would } be happy to have a order of magnitude. Is this 10 nm } or 2 µm deep? } Lets say I analyze at 15kV (1) NaCl crystals (2) } Quartz (3) Feldpar (4) Copper, what would be the } expected "depth of analysis" in these samples? } Does it depend on the size of the beam? } } Best regards, } } Stephane } } } ==============================Original Headers============================== } 14, 32 -- From wesaia-at-iastate.edu Tue May 22 14:01:16 2007 } 14, 32 -- Received: from mailhub-3.iastate.edu (mailhub-3.iastate.edu [129.186.140.13]) } 14, 32 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4MJ1GKq014627 } 14, 32 -- for {Microscopy-at-msa.microscopy.com} ; Tue, 22 May 2007 14:01:16 -0500 } 14, 32 -- Received: from mailout-2.iastate.edu (mailout-2.iastate.edu [129.186.140.2]) } 14, 32 -- by mailhub-3.iastate.edu (8.12.11.20060614/8.12.10) with SMTP id l4MJ1Fgu010242; } 14, 32 -- Tue, 22 May 2007 14:01:15 -0500 } 14, 32 -- Received: from (owa.eng.iastate.edu [129.186.23.85]) by mailout-2.iastate.edu with smtp } 14, 32 -- id 5cd8_6f1d9afc_0896_11dc_8987_001372578af6; } 14, 32 -- Tue, 22 May 2007 13:58:31 -0500 } 14, 32 -- Received: from maire.eng.iastate.edu ([10.10.196.69]) by owa.eng.iastate.edu with Microsoft SMTPSVC(6.0.3790.3959); } 14, 32 -- Tue, 22 May 2007 14:01:16 -0500 } 14, 32 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 } 14, 32 -- Content-class: urn:content-classes:message } 14, 32 -- MIME-Version: 1.0 } 14, 32 -- Content-Type: text/plain; } 14, 32 -- charset="iso-8859-1" } 14, 32 -- Subject: RE: [Microscopy] basic SEM: imaging salt crystals } 14, 32 -- Date: Tue, 22 May 2007 14:02:12 -0500 } 14, 32 -- Message-ID: {16A330AC32056A40B32842EC4BB8D72701A17957-at-maire.eng.iastate.edu} } 14, 32 -- In-Reply-To: {200705220947.l4M9lGXG018658-at-ns.microscopy.com} } 14, 32 -- X-MS-Has-Attach: } 14, 32 -- X-MS-TNEF-Correlator: } 14, 32 -- Thread-Topic: [Microscopy] basic SEM: imaging salt crystals } 14, 32 -- Thread-Index: AcecVi/FQpp2YuiQSJyH5uE2Kaj7bAASoivg } 14, 32 -- References: {200705220947.l4M9lGXG018658-at-ns.microscopy.com} } 14, 32 -- From: "Straszheim, Warren E [CCE E]" {wesaia-at-iastate.edu} } 14, 32 -- To: "MSA" {Microscopy-at-msa.microscopy.com} } 14, 32 -- Cc: {nizets2-at-yahoo.com} } 14, 32 -- X-OriginalArrivalTime: 22 May 2007 19:01:16.0474 (UTC) FILETIME=[931779A0:01C79CA3] } 14, 32 -- Content-Transfer-Encoding: 8bit } 14, 32 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4MJ1GKq014627 } ==============================End of - Headers============================== }
==============================Original Headers============================== 11, 29 -- From jacques.faerber-at-ipcms.u-strasbg.fr Wed May 23 11:11:50 2007 11, 29 -- Received: from mailhost.u-strasbg.fr (mailhost.u-strasbg.fr [130.79.200.158]) 11, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4NGBoaZ009690 11, 29 -- for {microscopy-at-microscopy.com} ; Wed, 23 May 2007 11:11:50 -0500 11, 29 -- Received: from ipcms.u-strasbg.fr (ipcms.u-strasbg.fr [130.79.210.2]) 11, 29 -- by mailhost.u-strasbg.fr (8.13.8/jtpda-5.5pre1) with ESMTP id l4NGBlF3077978 11, 29 -- for {microscopy-at-microscopy.com} ; Wed, 23 May 2007 18:11:47 +0200 (CEST) 11, 29 -- Received: from [130.79.152.3] (odhinn.u-strasbg.fr [130.79.152.3]) 11, 29 -- by ipcms.u-strasbg.fr (Postfix) with ESMTP id F07283EC01C 11, 29 -- for {Microscopy-at-Microscopy.Com} ; Wed, 23 May 2007 18:08:30 +0200 (CEST) 11, 29 -- Message-ID: {4654670E.7030702-at-ipcms.u-strasbg.fr} 11, 29 -- Date: Wed, 23 May 2007 18:08:46 +0200 11, 29 -- From: "j.faerber" {jacques.faerber-at-ipcms.u-strasbg.fr} 11, 29 -- User-Agent: Thunderbird 1.5.0.10 (X11/20070306) 11, 29 -- MIME-Version: 1.0 11, 29 -- To: Microscopy-at-microscopy.com 11, 29 -- Subject: Re: [Microscopy] RE: basic SEM: imaging salt crystals 11, 29 -- References: {200705221906.l4MJ6UPQ023853-at-ns.microscopy.com} 11, 29 -- In-Reply-To: {200705221906.l4MJ6UPQ023853-at-ns.microscopy.com} 11, 29 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 11, 29 -- Content-Transfer-Encoding: 8bit 11, 29 -- X-IPCMS-MailScanner: Found to be clean 11, 29 -- X-IPCMS-MailScanner-From: jacques.faerber-at-ipcms.u-strasbg.fr 11, 29 -- X-Greylist: Sender IP whitelisted, not delayed by milter-greylist-3.0 (mailhost.u-strasbg.fr [130.79.200.158]); Wed, 23 May 2007 18:11:48 +0200 (CEST) 11, 29 -- X-Virus-Scanned: ClamAV 0.88.7/3286/Wed May 23 14:11:14 2007 on mr8.u-strasbg.fr 11, 29 -- X-Virus-Status: Clean 11, 29 -- X-Spam-Status: No, score=-0.3 required=5.0 tests=AWL autolearn=disabled 11, 29 -- version=3.1.8 11, 29 -- X-Spam-Checker-Version: SpamAssassin 3.1.8 (2007-02-13) on mr8.u-strasbg.fr ==============================End of - Headers==============================
What is a "sample coating option?" I don't see a need for this. Just don't tag the coating peak(s) with their element nomenclature. Why one would want to quant NaCl is another and separate issue.
An 8KV beam on thin coated NaCl will produce nice pix and generate the desired peaks (slowly). With a Zeiss Supra 55VP, 30u aperture, low current, 10KV, the crystals do not melt. If one is worried about it, use smaller aperture and squeeze down the beam if your SEM is set up that way.
gary g.
At 08:13 AM 5/23/2007, you wrote:
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==============================Original Headers============================== 13, 22 -- From gary-at-gaugler.com Wed May 23 11:27:32 2007 13, 22 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 13, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l4NGRVot021447 13, 22 -- for {microscopy-at-microscopy.com} ; Wed, 23 May 2007 11:27:32 -0500 13, 22 -- Message-Id: {200705231627.l4NGRVot021447-at-ns.microscopy.com} 13, 22 -- Received: (qmail 24794 invoked from network); 23 May 2007 09:27:30 -0700 13, 22 -- Received: by simscan 1.1.0 ppid: 24791, pid: 24792, t: 0.2779s 13, 22 -- scanners: regex: 1.1.0 attach: 1.1.0 13, 22 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 13, 22 -- by qsmtp3 with SMTP; 23 May 2007 09:27:30 -0700 13, 22 -- X-Mailer: QUALCOMM Windows Eudora Version 7.1.0.9 13, 22 -- Date: Wed, 23 May 2007 09:27:26 -0800 13, 22 -- To: jacques.faerber-at-ipcms.u-strasbg.fr 13, 22 -- From: Gary Gaugler {gary-at-gaugler.com} 13, 22 -- Subject: Re: [Microscopy] basic SEM: imaging salt crystals 13, 22 -- Cc: MSA listserver {microscopy-at-microscopy.com} 13, 22 -- In-Reply-To: {200705231613.l4NGDZ2q012512-at-ns.microscopy.com} 13, 22 -- References: {200705231613.l4NGDZ2q012512-at-ns.microscopy.com} 13, 22 -- Mime-Version: 1.0 13, 22 -- Content-Type: text/plain; charset=iso-8859-1; format=flowed; x-avg-checked=avg-ok-6150735E 13, 22 -- Content-Transfer-Encoding: 8bit 13, 22 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4NGRVot021447 ==============================End of - Headers==============================
Mary Ellen, The rate that Walter suggested seemed extremely high compared to my experience with smaller animals so I asked an anatomist with many decades of experience with monkeys. He has always used an initial flush of 500ml in 5 minutes time with buffer followed by 1.5 liters of fixative over 20 minutes of time for a typical 5kg animal. The appropriate size tubing and cannula in collaboration with the setting on the pump will enable an excellent fixation. Walter's method may well provide good results. I would love to see some brain slices and TEM micrographs at 10,000X. I'm open to different methods.
Larry
Walter.Bobrowski-at-pfizer.com wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } 300 ml/min. Perfuse either through the carotid artery or left ventricle } (occlude descending aorta). } } Best regards, } Walter F. Bobrowski } Sr. Scientist } Pfizer Global R&D } 2800 Plymouth Rd. } Ann Arbor, MI 48105 } } TEL: 734-622-7814 } FAX: 734-622-5718 } } "Like the strength of a steel rod, the true character of a person can } only be known under extreme stress." - Leslie Fieger } } } } } -----Original Message----- } X-from: mpease-at-jhmi.edu [mailto:mpease-at-jhmi.edu] } Sent: Monday, May 21, 2007 7:25 PM } To: Bobrowski, Walter } Subject: [Microscopy] viaWWW: monkey perfusion methodology } } } } } ------------------------------------------------------------------------ } ---- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ } ---- } } This Question/Comment was submitted to the Microscopy Listserver } using the WWW based Form at } http://www.microscopy.org/MicroscopyListserver/MLFormMail.html } ------------------------------------------------------------------------ } --- } Remember this posting is most likely not from a Subscriber, so when } replying } please copy both mpease-at-jhmi.edu as well as the MIcroscopy } Listserver } ------------------------------------------------------------------------ } --- } } Email: mpease-at-jhmi.edu } Name: Mary Ellen Pease } } Organization: Johns Hopkins Wilmer Eye Institute } } Title-Subject: [Filtered] monkey perfusion methodology } } Question: What is the recommended flow rate for perfusion of monkey } brain and eyes when using a pump rather than gravity feed of the } solutions? } } Thank you, } Mary Ellen } } ------------------------------------------------------------------------ } --- } } ==============================Original } Headers============================== } 7, 11 -- From zaluzec-at-microscopy.com Mon May 21 18:19:37 2007 } 7, 11 -- Received: from [206.69.208.22] (mac22.zaluzec.com } [206.69.208.22]) } 7, 11 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id l4LNJbJx007966 } 7, 11 -- for {microscopy-at-microscopy.com} ; Mon, 21 May 2007 } 18:19:37 -0500 } 7, 11 -- Mime-Version: 1.0 } 7, 11 -- Message-Id: {p06240806c277d979e099-at-[206.69.208.22]} } 7, 11 -- Date: Mon, 21 May 2007 18:19:36 -0500 } 7, 11 -- To: microscopy-at-microscopy.com } 7, 11 -- From: mpease-at-jhmi.edu (by way of MicroscopyListserver) } 7, 11 -- Subject: viaWWW: monkey perfusion methodology } 7, 11 -- Content-Type: text/plain; charset="us-ascii" } ==============================End of - } Headers============================== } } ---------------------------------------------------------------------- } LEGAL NOTICE } Unless expressly stated otherwise, this message is confidential and may be privileged. 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-- Larry Ackerman, Associate Specialist UCSF, Dept. of Anatomy, Rm S1347 513 Parnassus Ave., Box 0452 San Francisco, CA 94143
larry.ackerman-at-ucsf.edu
==============================Original Headers============================== 6, 28 -- From Larry.Ackerman-at-ucsf.edu Wed May 23 12:41:45 2007 6, 28 -- Received: from emfmcb02.ucsfmedicalcenter.org (emfmcb02.ucsfmedicalcenter.org [64.54.46.98]) 6, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4NHfijF002378 6, 28 -- for {Microscopy-at-microscopy.com} ; Wed, 23 May 2007 12:41:44 -0500 6, 28 -- Received: from 64.54.128.152 by emfmcb01.ucsfmedicalcenter.org with 6, 28 -- ESMTP (Tumbleweed Email Firewall SMTP Relay (Email Firewall v6.1.0)); 6, 28 -- Wed, 23 May 2007 10:53:00 -0700 6, 28 -- X-Server-Uuid: E2E48A14-EE5B-4280-A138-188440602EDD 6, 28 -- Received: from baluk.ucsf.edu ([128.218.123.88]) by exvs06.net.ucsf.edu 6, 28 -- with Microsoft SMTPSVC(6.0.3790.3959); Wed, 23 May 2007 10:41:19 -0700 6, 28 -- Message-ID: {46547CBF.8050702-at-ucsf.edu} 6, 28 -- Date: Wed, 23 May 2007 10:41:19 -0700 6, 28 -- From: "Larry Ackerman" {larry.ackerman-at-ucsf.edu} 6, 28 -- Reply-to: larry.ackerman-at-ucsf.edu 6, 28 -- Organization: UCSF, NeuroAnatomy 6, 28 -- User-Agent: Thunderbird 2.0.0.0 (Macintosh/20070326) 6, 28 -- MIME-Version: 1.0 6, 28 -- To: mpease-at-jhmi.edu, Microscopy-at-microscopy.com 6, 28 -- Subject: Re: [Microscopy] monkey perfusion methodology 6, 28 -- References: {200705221231.l4MCVVEO018334-at-ns.microscopy.com} 6, 28 -- In-Reply-To: {200705221231.l4MCVVEO018334-at-ns.microscopy.com} 6, 28 -- X-OriginalArrivalTime: 23 May 2007 17:41:19.0789 (UTC) 6, 28 -- FILETIME=[92752DD0:01C79D61] 6, 28 -- X-WSS-ID: 6A4AA0F00MC2362538-15-01 6, 28 -- Content-Type: text/plain; 6, 28 -- charset=iso-8859-1; 6, 28 -- format=flowed 6, 28 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Dear All, can somebody help me out with the name and species of a tiny little animal I found on an absinthe leaf? See http://www.elektronenmikroskopie.info/absinthe_animal.jpg
Websites: www.stefan-diller.com www.elektronenmikroskopie.info www.assisi.de Anfahrt: http://mail.map24.com/stefan.diller ---------------------------------------------------------------------------- ----------------------------------------- ----- Original Message ----- X-from: {gary-at-gaugler.com} To: {stefan.diller-at-t-online.de} Sent: Wednesday, May 23, 2007 6:33 PM
Stefan,
Do you have any more images? Different angles of view would help
Phil
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==============================Original Headers============================== 4, 22 -- From oshel1pe-at-cmich.edu Wed May 23 14:40:35 2007 4, 22 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4NJeZQG028163 4, 22 -- for {Microscopy-at-microscopy.com} ; Wed, 23 May 2007 14:40:35 -0500 4, 22 -- Received: from egatea.central.cmich.local ([141.209.15.74]) 4, 22 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id l4NK3Wqu023520 4, 22 -- for {Microscopy-at-microscopy.com} ; Wed, 23 May 2007 16:03:36 -0400 4, 22 -- Received: from [141.209.160.249] ([141.209.160.249]) by egatea.central.cmich.local with Microsoft SMTPSVC(6.0.3790.1830); 4, 22 -- Wed, 23 May 2007 15:40:28 -0400 4, 22 -- Mime-Version: 1.0 4, 22 -- Message-Id: {f0623090fc27a48cc5e95-at-[141.209.160.249]} 4, 22 -- In-Reply-To: {200705231913.l4NJDUWD022580-at-ns.microscopy.com} 4, 22 -- References: {200705231913.l4NJDUWD022580-at-ns.microscopy.com} 4, 22 -- Date: Wed, 23 May 2007 15:40:26 -0400 4, 22 -- To: Microscopy-at-microscopy.com 4, 22 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 4, 22 -- Subject: Re: [Microscopy] Unknown creature 4, 22 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 4, 22 -- X-OriginalArrivalTime: 23 May 2007 19:40:28.0770 (UTC) FILETIME=[37951C20:01C79D72] 4, 22 -- X-CanItPRO-Stream: default 4, 22 -- X-Spam-Score: -3.5 () INFO_TLD,L_EXCH_MF 4, 22 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
Our nanoscience program received some information about the Phenom-Ed from FEI, a tablettop SEM designed for education. We are considering whether it would useful for students to perform quick analysis of nanomaterials. Sounds like the instrument is still in the prototype stage, but we are wondering if anyone has any impressions they could share on this. From the price we were quoted, it might actually be within our budget.
Some info on this product is available at: http://investor.fei.com/phoenix.zhtml?c=60978&p=irol-newsArticle&ID=9682 99&highlight=
Thanks in advance for your feedback. -Phil ------------------------------------------ Phil Ahrenkiel, Assistant Professor Nanoscience and Nanoengineering Ph.D. Program South Dakota School of Mines and Technology 501 East St. Joseph Street Rapid City, South Dakota 57701 Office: EEP 221 Phone: (605)394-5238, Fax: (605)394-2365 Email: Phil.Ahrenkiel-at-sdsmt.edu
==============================Original Headers============================== 7, 20 -- From Phil.Ahrenkiel-at-sdsmt.edu Wed May 23 14:49:01 2007 7, 20 -- Received: from owa.sdsmt.edu (owa.sdsmt.edu [151.159.3.7]) 7, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4NJn0OO007452 7, 20 -- for {Microscopy-at-microscopy.com} ; Wed, 23 May 2007 14:49:00 -0500 7, 20 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 7, 20 -- Content-class: urn:content-classes:message 7, 20 -- MIME-Version: 1.0 7, 20 -- Content-Type: text/plain; 7, 20 -- charset="us-ascii" 7, 20 -- Subject: Phenom-Ed SEM 7, 20 -- Date: Wed, 23 May 2007 13:48:59 -0600 7, 20 -- Message-ID: {6FD01225FACD9C4AA3870724330536F4065B5B9F-at-sdsmt-ex01.SDSMT.LOCAL} 7, 20 -- X-MS-Has-Attach: 7, 20 -- X-MS-TNEF-Correlator: 7, 20 -- Thread-Topic: Phenom-Ed SEM 7, 20 -- thread-index: Acedc2ffJOT9yU/VTH23gIJB3edmtA== 7, 20 -- From: "Ahrenkiel, Scott \(Phil\) P." {Phil.Ahrenkiel-at-sdsmt.edu} 7, 20 -- To: {Microscopy-at-microscopy.com} 7, 20 -- Content-Transfer-Encoding: 8bit 7, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4NJn0OO007452 ==============================End of - Headers==============================
I would not call it "prototype stage" anymore. As coincidence would have it, I was at the official introduction of the Phenom at the Fontys University of Applied Sciences in Eindhoven, The Netherlands, earlier today. The instrument will be used there exactly for what you suggest.
There is (much) more information available at http:// {http://www.fei.com/phenom} www.fei.com/phenom.
I won't say more about the instrument here on the list (unless requested) since I am part of the team who actually designed it, and hence would come across as rather biased (in a positive way!). There are links on the website with contact information. Please click on them, we love talking about it!
Our nanoscience program received some information about the Phenom-Ed from FEI, a tablettop SEM designed for education. We are considering whether it would useful for students to perform quick analysis of nanomaterials. Sounds like the instrument is still in the prototype stage, but we are wondering if anyone has any impressions they could share on this. From the price we were quoted, it might actually be within our budget.
Some info on this product is available at: http://investor.fei.com/phoenix.zhtml?c=60978&p=irol-newsArticle&ID=9682 99&highlight=
Thanks in advance for your feedback. -Phil ------------------------------------------ Phil Ahrenkiel, Assistant Professor Nanoscience and Nanoengineering Ph.D. Program South Dakota School of Mines and Technology 501 East St. Joseph Street Rapid City, South Dakota 57701 Office: EEP 221 Phone: (605)394-5238, Fax: (605)394-2365 Email: Phil.Ahrenkiel-at-sdsmt.edu
==============================Original Headers============================== 7, 20 -- From Phil.Ahrenkiel-at-sdsmt.edu Wed May 23 14:49:01 2007 7, 20 -- Received: from owa.sdsmt.edu (owa.sdsmt.edu [151.159.3.7]) 7, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4NJn0OO007452 7, 20 -- for {Microscopy-at-microscopy.com} ; Wed, 23 May 2007 14:49:00 -0500 7, 20 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 7, 20 -- Content-class: urn:content-classes:message 7, 20 -- MIME-Version: 1.0 7, 20 -- Content-Type: text/plain; 7, 20 -- charset="us-ascii" 7, 20 -- Subject: Phenom-Ed SEM 7, 20 -- Date: Wed, 23 May 2007 13:48:59 -0600 7, 20 -- Message-ID: {6FD01225FACD9C4AA3870724330536F4065B5B9F-at-sdsmt-ex01.SDSMT.LOCAL} 7, 20 -- X-MS-Has-Attach: 7, 20 -- X-MS-TNEF-Correlator: 7, 20 -- Thread-Topic: Phenom-Ed SEM 7, 20 -- thread-index: Acedc2ffJOT9yU/VTH23gIJB3edmtA== 7, 20 -- From: "Ahrenkiel, Scott \(Phil\) P." {Phil.Ahrenkiel-at-sdsmt.edu} 7, 20 -- To: {Microscopy-at-microscopy.com} 7, 20 -- Content-Transfer-Encoding: 8bit 7, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4NJn0OO007452 ==============================End of - Headers==============================
==============================Original Headers============================== 20, 28 -- From Sander.Stoks-at-fei.com Wed May 23 15:30:29 2007 20, 28 -- Received: from smtp-nl1.feico.com (smtp-nl1.feico.com [195.75.179.210]) 20, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4NKUTu7019853 20, 28 -- for {microscopy-at-microscopy.com} ; Wed, 23 May 2007 15:30:29 -0500 20, 28 -- X-WSS-ID: 0JIIGYZ-02-UHC-01 20, 28 -- Received: from acht850.w2k.feico.com (unknown [10.150.55.50]) 20, 28 -- by smtp-nl1.feico.com (Tumbleweed MailGate) with SMTP id 51310110759F 20, 28 -- for {microscopy-at-microscopy.com} ; Wed, 23 May 2007 13:30:35 -0700 (PDT) 20, 28 -- Received: From acht887.w2k.feico.com ([10.150.55.87]) by acht850.w2k.feico.com (WebShield SMTP v4.5 MR2); 20, 28 -- id 1179952225849; Wed, 23 May 2007 22:30:25 +0200 20, 28 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 20, 28 -- Content-class: urn:content-classes:message 20, 28 -- MIME-Version: 1.0 20, 28 -- Content-Type: text/plain; 20, 28 -- charset="iso-8859-1" 20, 28 -- Subject: RE: [Microscopy] Phenom-Ed SEM 20, 28 -- Date: Wed, 23 May 2007 22:30:25 +0200 20, 28 -- Message-ID: {9211E4E38C8130428654AD6A1081A2CD0BF4F07C-at-acht887.w2k.feico.com} 20, 28 -- X-MS-Has-Attach: 20, 28 -- X-MS-TNEF-Correlator: 20, 28 -- Thread-Topic: [Microscopy] Phenom-Ed SEM 20, 28 -- Thread-Index: AceddAE622YIKLjISOuVfjEXX01ykAAAiDtl 20, 28 -- References: {200705231953.l4NJrAf7017176-at-ns.microscopy.com} 20, 28 -- From: "Stoks, Sander" {Sander.Stoks-at-fei.com} 20, 28 -- To: {microscopy-at-microscopy.com} 20, 28 -- Cc: {Phil.Ahrenkiel-at-sdsmt.edu} 20, 28 -- Content-Transfer-Encoding: 8bit 20, 28 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4NKUTu7019853 ==============================End of - Headers==============================
} From mail-at-ns.microscopy.com Wed May 23 15:48:05 2007 } Date: Wed, 23 May 2007 14:49:29 -0500 } To: jquinn-at-www.matscieng.sunysb.edu } From: Phil.Ahrenkiel-at-sdsmt.edu } Reply-to: Phil.Ahrenkiel-at-sdsmt.edu } X-Resent-From: "Microscopy Listserver" {microscopy-at-microscopy.com} } Subject: [Microscopy] Phenom-Ed SEM } Errors-To: MicroscopyListSpamFilter-at-microscopy.com } X-lewp: MicroscopyListSpam NAGS } } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } } Greetings- } } Our nanoscience program received some information about the Phenom-Ed } from FEI, a tablettop SEM designed for education. We are considering } whether it would useful for students to perform quick analysis of } nanomaterials. Sounds like the instrument is still in the prototype } stage, but we are wondering if anyone has any impressions they could } share on this. From the price we were quoted, it might actually be } within our budget. } } Some info on this product is available at: } http://investor.fei.com/phoenix.zhtml?c=60978&p=irol-newsArticle&ID=9682 } 99&highlight= } } Thanks in advance for your feedback. } -Phil } ------------------------------------------ } Phil Ahrenkiel, Assistant Professor } Nanoscience and Nanoengineering Ph.D. Program } South Dakota School of Mines and Technology } 501 East St. Joseph Street } Rapid City, South Dakota 57701 } Office: EEP 221 } Phone: (605)394-5238, Fax: (605)394-2365 } Email: Phil.Ahrenkiel-at-sdsmt.edu } } } } ==============================Original Headers============================== } 7, 20 -- From Phil.Ahrenkiel-at-sdsmt.edu Wed May 23 14:49:01 2007 } 7, 20 -- Received: from owa.sdsmt.edu (owa.sdsmt.edu [151.159.3.7]) } 7, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4NJn0OO007452 } 7, 20 -- for {Microscopy-at-microscopy.com} ; Wed, 23 May 2007 14:49:00 -0500 } 7, 20 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 } 7, 20 -- Content-class: urn:content-classes:message } 7, 20 -- MIME-Version: 1.0 } 7, 20 -- Content-Type: text/plain; } 7, 20 -- charset="us-ascii" } 7, 20 -- Subject: Phenom-Ed SEM } 7, 20 -- Date: Wed, 23 May 2007 13:48:59 -0600 } 7, 20 -- Message-ID: {6FD01225FACD9C4AA3870724330536F4065B5B9F-at-sdsmt-ex01.SDSMT.LOCAL} } 7, 20 -- X-MS-Has-Attach: } 7, 20 -- X-MS-TNEF-Correlator: } 7, 20 -- Thread-Topic: Phenom-Ed SEM } 7, 20 -- thread-index: Acedc2ffJOT9yU/VTH23gIJB3edmtA== } 7, 20 -- From: "Ahrenkiel, Scott \(Phil\) P." {Phil.Ahrenkiel-at-sdsmt.edu} } 7, 20 -- To: {Microscopy-at-microscopy.com} } 7, 20 -- Content-Transfer-Encoding: 8bit } 7, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4NJn0OO007452 } ==============================End of - Headers============================== }
Phil and others -
Tell me the cost and I will give you an answer.
It certainly looks like a great entry level unit.
But a few caveats.......
1) I would question the use of the word "nanoscale".
2) You will never get EDS out of it.
3) You cannot use large samples.
The last two would exclude 3/4 of my users.
However, I eagerly look forward to the cost.
kind regards,
Jim
PS: OoO away.........
} From mail-at-ns.microscopy.com Wed May 23 15:48:05 2007 } Date: Wed, 23 May 2007 14:49:29 -0500 } To: jquinn-at-www.matscieng.sunysb.edu } From: Phil.Ahrenkiel-at-sdsmt.edu } Reply-to: Phil.Ahrenkiel-at-sdsmt.edu } X-Resent-From: "Microscopy Listserver" {microscopy-at-microscopy.com} } Subject: [Microscopy] Phenom-Ed SEM } Errors-To: MicroscopyListSpamFilter-at-microscopy.com } X-lewp: MicroscopyListSpam NAGS } } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } } Greetings- } } Our nanoscience program received some information about the Phenom-Ed } from FEI, a tablettop SEM designed for education. We are considering } whether it would useful for students to perform quick analysis of } nanomaterials. Sounds like the instrument is still in the prototype } stage, but we are wondering if anyone has any impressions they could } share on this. From the price we were quoted, it might actually be } within our budget. } } Some info on this product is available at: } http://investor.fei.com/phoenix.zhtml?c=60978&p=irol-newsArticle&ID=9682 } 99&highlight= } } Thanks in advance for your feedback. } -Phil } ------------------------------------------ } Phil Ahrenkiel, Assistant Professor } Nanoscience and Nanoengineering Ph.D. Program } South Dakota School of Mines and Technology } 501 East St. Joseph Street } Rapid City, South Dakota 57701 } Office: EEP 221 } Phone: (605)394-5238, Fax: (605)394-2365 } Email: Phil.Ahrenkiel-at-sdsmt.edu } } } } ==============================Original Headers============================== } 7, 20 -- From Phil.Ahrenkiel-at-sdsmt.edu Wed May 23 14:49:01 2007 } 7, 20 -- Received: from owa.sdsmt.edu (owa.sdsmt.edu [151.159.3.7]) } 7, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4NJn0OO007452 } 7, 20 -- for {Microscopy-at-microscopy.com} ; Wed, 23 May 2007 14:49:00 -0500 } 7, 20 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 } 7, 20 -- Content-class: urn:content-classes:message } 7, 20 -- MIME-Version: 1.0 } 7, 20 -- Content-Type: text/plain; } 7, 20 -- charset="us-ascii" } 7, 20 -- Subject: Phenom-Ed SEM } 7, 20 -- Date: Wed, 23 May 2007 13:48:59 -0600 } 7, 20 -- Message-ID: {6FD01225FACD9C4AA3870724330536F4065B5B9F-at-sdsmt-ex01.SDSMT.LOCAL} } 7, 20 -- X-MS-Has-Attach: } 7, 20 -- X-MS-TNEF-Correlator: } 7, 20 -- Thread-Topic: Phenom-Ed SEM } 7, 20 -- thread-index: Acedc2ffJOT9yU/VTH23gIJB3edmtA== } 7, 20 -- From: "Ahrenkiel, Scott \(Phil\) P." {Phil.Ahrenkiel-at-sdsmt.edu} } 7, 20 -- To: {Microscopy-at-microscopy.com} } 7, 20 -- Content-Transfer-Encoding: 8bit } 7, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4NJn0OO007452 } ==============================End of - Headers============================== }
==============================Original Headers============================== 15, 13 -- From jquinn-at-www.matscieng.sunysb.edu Wed May 23 18:58:54 2007 15, 13 -- Received: from www.matscieng.sunysb.edu (www.matscieng.sunysb.edu [129.49.36.33]) 15, 13 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4NNwslg003573 15, 13 -- for {microscopy-at-microscopy.com} ; Wed, 23 May 2007 18:58:54 -0500 15, 13 -- Received: (from jquinn-at-localhost) 15, 13 -- by www.matscieng.sunysb.edu (8.11.6/8.11.6) id l4NNvOe20702; 15, 13 -- Wed, 23 May 2007 19:57:24 -0400 15, 13 -- Date: Wed, 23 May 2007 19:57:24 -0400 15, 13 -- From: Jim Quinn {jquinn-at-www.matscieng.sunysb.edu} 15, 13 -- Message-Id: {200705232357.l4NNvOe20702-at-www.matscieng.sunysb.edu} 15, 13 -- To: microscopy-at-microscopy.com 15, 13 -- Subject: Re: [Microscopy] Phenom-Ed SEM 15, 13 -- Cc: Phil.Ahrenkiel-at-sdsmt.edu ==============================End of - Headers==============================
jquinn-at-www.matscieng.sunysb.edu wrote: } Phil and others - } } Tell me the cost and I will give you an answer. } It certainly looks like a great entry level unit. } But a few caveats....... } 1) I would question the use of the word "nanoscale". } 2) You will never get EDS out of it. } 3) You cannot use large samples. } The last two would exclude 3/4 of my users. } However, I eagerly look forward to the cost. } kind regards, } Jim } PS: OoO away......... }
Well, 62,000 Euro is the RRP from the website.
-- Scott J. Coutts ------------------------------------------------------------------------- Bacterial Pathogenesis Research Group Department of Microbiology Box 53, Monash University, 3800, Australia Phone +61 3 9905 8592, Fax +61 3 9905 4811 -------------------------------------------------------------------------
==============================Original Headers============================== 4, 28 -- From scott.coutts-at-med.monash.edu.au Thu May 24 01:11:05 2007 4, 28 -- Received: from kenny.its.monash.edu.au (kenny.its.monash.edu.au [130.194.13.164]) 4, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4O6B4dR020940 4, 28 -- for {microscopy-at-microscopy.com} ; Thu, 24 May 2007 01:11:05 -0500 4, 28 -- Received: from moe.its.monash.edu.au ([130.194.13.88]) 4, 28 -- by kenny.its.monash.edu.au 4, 28 -- (Sun Java System Messaging Server 6.2-4.03 (built Sep 22 2005)) 4, 28 -- with ESMTP id {0JIJ00HJW7UETLB0-at-kenny.its.monash.edu.au} for 4, 28 -- microscopy-at-microscopy.com; Thu, 24 May 2007 16:11:03 +1000 (EST) 4, 28 -- Received: from moe.its.monash.edu.au (localhost.localdomain [127.0.0.1]) 4, 28 -- by localhost (Postfix) with ESMTP id 585BEAB542 for 4, 28 -- {microscopy-at-microscopy.com} ; Thu, 24 May 2007 16:11:03 +1000 (EST) 4, 28 -- Received: from [130.194.200.122] 4, 28 -- (MicrobSCouttsDT.med.monash.edu.au [130.194.200.122]) 4, 28 -- by moe.its.monash.edu.au (Postfix) with ESMTP id 3A3254FB07 for 4, 28 -- {microscopy-at-microscopy.com} ; Thu, 24 May 2007 16:11:03 +1000 (EST) 4, 28 -- Date: Thu, 24 May 2007 16:11:36 +1000 4, 28 -- From: Scott Coutts {scott.coutts-at-med.monash.edu.au} 4, 28 -- Subject: Re: [Microscopy] Re: Phenom-Ed SEM 4, 28 -- In-reply-to: {200705240002.l4O02orv009494-at-ns.microscopy.com} 4, 28 -- To: microscopy-at-microscopy.com 4, 28 -- Message-id: {46552C98.30406-at-med.monash.edu.au} 4, 28 -- Organization: Monash University 4, 28 -- MIME-version: 1.0 4, 28 -- Content-type: text/plain; charset=ISO-8859-1; format=flowed 4, 28 -- Content-transfer-encoding: 7bit 4, 28 -- References: {200705240002.l4O02orv009494-at-ns.microscopy.com} 4, 28 -- User-Agent: Thunderbird 1.5.0.10 (Windows/20070221) ==============================End of - Headers==============================
} Our nanoscience program received some information about the } Phenom-Ed from FEI, a tablettop SEM designed for education. } We are considering whether it would useful for students to } perform quick analysis of nanomaterials. ...
Regarding "nanomaterials", keep in mind that magnifications are not what they used to be. For example, it used to be that a SEM's mag number was based on a 4x5 polaroid, whereas now the mag number is based on a 10" window on the computer display. The white paper describes the Phenom capable of mag=20,000x which, relative to SEMs we are used to, would equate to 10,000x (1 cm = 1,000 nM on a 4x5).
Not to take away from what should be a very valuable and interesting tool in a science program for many high schools and colleges, and I only comment relative to the term "nanomaterials".
cheerios :o) michael shaffer
SEM/MLA Research Coordinator {http://www.mun.ca/creait/maf/} Inco Innovation Centre Memorial University St. John's, Newfoundland
==============================Original Headers============================== 7, 20 -- From michael-at-shaffer.net Thu May 24 06:36:19 2007 7, 20 -- Received: from n007.sc1.he.tucows.com (smtpout1446.sc1.he.tucows.com [64.97.157.146]) 7, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4OBaJp4007503 7, 20 -- for {Microscopy-at-microscopy.com} ; Thu, 24 May 2007 06:36:19 -0500 7, 20 -- Received: from roamingwolf (134.153.130.141) by n007.sc1.he.tucows.com (7.2.069.1) (authenticated as michael-at-shaffer.net) 7, 20 -- id 4630CC6B004AED22 for Microscopy-at-microscopy.com; Thu, 24 May 2007 11:36:18 +0000 7, 20 -- From: "michael shaffer" {michael-at-shaffer.net} 7, 20 -- To: "MSA Microscopy list" {Microscopy-at-microscopy.com} 7, 20 -- References: {200705231949.l4NJnIjx008103-at-ns.microscopy.com} 7, 20 -- Subject: RE: [Microscopy] Phenom-Ed SEM 7, 20 -- Date: Thu, 24 May 2007 09:09:09 -0230 7, 20 -- Message-ID: {000f01c79df8$2569cf10$8d829986-at-CREAIT.MUN.CA} 7, 20 -- MIME-Version: 1.0 7, 20 -- Content-Type: text/plain; 7, 20 -- charset="US-ASCII" 7, 20 -- Content-Transfer-Encoding: 7bit 7, 20 -- X-Mailer: Microsoft Office Outlook 11 7, 20 -- thread-index: AcedcuSS+ggTxhopQim0x4OAZEW60QAgrMaw 7, 20 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 7, 20 -- In-Reply-To: {200705231949.l4NJnIjx008103-at-ns.microscopy.com} ==============================End of - Headers==============================
Gary, in answer to your question, the last time I made a round robin to buy an EDS system (back 6 years ago or more), one manufacturer, but I don't remember who, had that option, in which you give your coating material and an estimated thickness. It takes the coating in account to correct the selective absorbtion of the X-ray lines from the sample by the coating. Perheps someone has informations on that possibility.
Of coarse, it has not much meaning to quantify NaCl by EDS (only to test your quanti software !). But, Na is a typical exemple of that problem : low energy lines will be absorbed selectively, in comparaison to higher energy lines. This particulary the case with O-k, l lines from transition metals, Na, Mg, etc. The quantification will not be correct, if you only ignore the presence of gold. The balance between Ca, K on one side, and Na or Mg (and O if one use the measured intensities) on the other side will be biased.
Of coarse, with 5 nm of a grainy gold coating (blue at daylight), where electrical conducitvity is done mostly by tunneling between gold islands, letting a large aera of the sample uncoated, you will not see a big error. But with a nice yellowish } 25 nm coating, which cover uniformely the sample, the effect can be very effective. I have recently seen pictures/samples with a uniform grainy surface from the coating, in a case where one was looking after Cl and S in a polymer. No hope to see something in such a case. And such situations are unfortunatly not unfrequent.
As said, Na is soon difficult to quantify correctly, in glasses for exemple. So I prefer to be carefull, and not add a source of errors.
Do you think I'm overcarefull ? Maybe !!!
jcqs
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
gary-at-gaugler.com a écrit : } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } What is a "sample coating option?" I don't see a need } for this. Just don't tag the coating peak(s) with their } element nomenclature. Why one would want to quant NaCl } is another and separate issue. } } An 8KV beam on thin coated NaCl will produce nice pix } and generate the desired peaks (slowly). With a Zeiss } Supra 55VP, 30u aperture, low current, 10KV, the crystals } do not melt. If one is worried about it, use smaller } aperture and squeeze down the beam if your SEM is set up } that way. } } gary g. } } } } } At 08:13 AM 5/23/2007, you wrote: } } } } } } } ---------------------------------------------------------------------------- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------------- } } } } Hi all } } } } One or two cent more! } } } } For EDS analysis, if you coat your crystals with a high Z material, and } } your analysis software dont' have an "sample coating" option, you'll } } probably will have wrong results on crystals like NaCl. It's always } } difficult to have accurate results on Na (it evaoprate under the beam, } } but not enough to polluate the SEM !), but with gold coating, absorption } } } } from the Na-k by the gold layer will be much stronger than that from } } } Cl-k, and the balance Na-Cl will be worse. I think that carbon is the } } less problematic coating for such analysis, and I would not use Au or } } Pt. Of coarse, imaging can be less nice than with Au, Pt or Ir coating. } } } } For Monte Carlo simulation, you can download Casino } } (http://www.gel.usherbrooke.ca/casino/), with has some nice possibility, } } like multiple energies simulation. I use it complementary to Electron } } Flight Simulator. } } } } A simple test in particule (or thin layer) analysis, is to put the } } particules on a substrate which don't have any element from the } } particules (of coarse, you must have some idees about these particules, } } and yes, you don't have any, that's what your are looking for !) and to } } perform measurement with different energies, from say 5 to 20 keV. As } } long as you see the x-ray lines of the subtrate in the spectrum, the } } primary energie is too hight. By the way, it is suffisent that one } } element from the substrate does not exist in the particule, if possible } } with a line at low energie, to be able to see it in the spectrum at all } } energies (Cu-L par exemple). } } } } Hope it helps } } } } J. Faerber } } IPCMS-GSI } } (Institut de Physique et Chimie des Matériaux de Strasbourg } } Groupe Surface et Interfaces) } } 23, rue de Loess ; BP43 } } 67034 Strasbourg CEDEX 2 } } France } } } } Tel 00 33(0)3 88 10 71 01 } } Fax 00 33(0)3 88 10 72 48 } } E-mail Jacques.Faerber-at-ipcms.u-strasbg.fr } } } } } } } } wesaia-at-iastate.edu a écrit : } } } } } } } ---------------------------------------------------------------------------- } } } } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } } } ---------------------------------------------------------------------------- } } } } } I have already seen a number of good answers, but I will chime in anyway. } } } } } } I would suggest coating the samples unless } } } } } you are operating in variable pressure mode } } (which may not be an option for you). I recall } } that NaCl charges as well as most any } } insulator. I don't imagine you would be doing } } quant on the sample since the lack of a known } } geometry would lead to substantial uncertainty } } in the results. Gary made a good point that it } } would be nice to use an element for coating } } that is only present in the coating. Otherwise, } } there will be uncertainty in the source of the } } peaks. I would expect some difference in the } } relative peak intensities depending on which element is used. } } } } } I wouldn't worry much about vaporizing the } } } } } salt. (Maybe I should.) I don't expect you will } } be using nanoamps of current. Once again, VP } } mode would help minimize or prevent contamination. } } } } } I don't have the latest and greatest Monte } } } } } Carlo programs. I do have an old one derived } } from David Joy's work. I ran a simulation on S } } at 15 kV and I estimate the excitation depth to } } be slightly less than 1 um. Running the same } } simulation for Na, I found the depth to be more } } like 2 um. For Cu, I found the depth to be on } } the order of 0.3 um. Therefore, I generally } } quote penetration depths of hundreds of nm up } } to a couple of microns. The dimensions of the } } interaction volume dwarf the spot size so much } } that I don't often bother considering it in calculating interaction volume. } } } } } As a fun little exercise, you might lay down } } } } } a gold layer on copper or some other metal. The } } interaction volume for gold appears to be } } slightly less than 0.2 um (200 nm). Since we } } routinely coat samples with 10 nm of gold, we } } need a deeper penetration depth if we are ever } } going to see the sample through the gold. We } } don't have a problem seeing the sample unless } } we are using a very thick layer of gold or a } } low voltage. A 3 kV beam would have difficulty } } penetrating 10 nm of gold, but there would not } } be much excitation of sample x-rays at that low a voltage anyway. } } } } } Warren } } } } } } -----Original Message----- } } } X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] } } } Sent: Tuesday, May 22, 2007 4:47 AM } } } To: wesaia-at-iastate.edu } } } Subject: [Microscopy] basic SEM: imaging salt crystals } } } } } } Dear listers, } } } } } } Here are some basic questions from the basic SEM user } } } I am: I would like to visualize salt crystals by SEM } } } and make EDX analysis if possible. I just leave a drop } } } of salt solution on a carbon tab dry, that's all! } } } My questions: } } } } } } - Do I need to carbon-coat the crystals? } } } - Is there a risk to evaporate the crystals under the } } } electron beam and make the column dirty? (I usually } } } don't use HV higher than 15 kV). Would carbon coating } } } help dissipate the heat in this regard? } } } } } } Now another question related to SEM but not to } } } crystals: } } } When I analyse fine particles by EDX, I never know if } } } I analyse only the particle or also part of the } } } material which is behind the particle. Is there a way } } } to determine the depth of analysis of the material } } } studied? I suppose that it depends on the HV and on } } } the nature of the material itself, but I already would } } } be happy to have a order of magnitude. Is this 10 nm } } } or 2 µm deep? } } } Lets say I analyze at 15kV (1) NaCl crystals (2) } } } Quartz (3) Feldpar (4) Copper, what would be the } } } expected "depth of analysis" in these samples? } } } Does it depend on the size of the beam? } } } } } } Best regards, } } } } } } Stephane } } } } } } } } } ==============================Original } } } } } Headers============================== } } } } } 14, 32 -- From wesaia-at-iastate.edu Tue May 22 14:01:16 2007 } } } 14, 32 -- Received: from } } } } } mailhub-3.iastate.edu (mailhub-3.iastate.edu [129.186.140.13]) } } } } } 14, 32 -- by ns.microscopy.com } } } } } (8.12.11.20060308/8.12.8) with ESMTP id l4MJ1GKq014627 } } } } } 14, 32 -- for } } } } } {Microscopy-at-msa.microscopy.com} ; Tue, 22 May 2007 14:01:16 -0500 } } } } } 14, 32 -- Received: from } } } } } mailout-2.iastate.edu (mailout-2.iastate.edu [129.186.140.2]) } } } } } 14, 32 -- by mailhub-3.iastate.edu } } } } } (8.12.11.20060614/8.12.10) with SMTP id l4MJ1Fgu010242; } } } } } 14, 32 -- Tue, 22 May 2007 14:01:15 -0500 } } } 14, 32 -- Received: from (owa.eng.iastate.edu } } } } } [129.186.23.85]) by mailout-2.iastate.edu with smtp } } } } } 14, 32 -- id 5cd8_6f1d9afc_0896_11dc_8987_001372578af6; 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Title-Subject: [Filtered] Re: [Microscopy] Phenom-Ed SEM
Question: Just to throw in another option. I saw a quick review of a Hitachi tabletop SEM. It's always good to see a comparison. I'm really not interested in either one. They are limited in function. Hitachi's model is BSE only and had limited stage manipulation capability. They maybe a good learning tool for someone who has never been exposed to an SEM before. It's not a replacement for a true SEM, just a fun toy.
I have an issue regarding perfusion fixed (glut & paraform. in cacodylate buffer) adult rat spinal cord.
I have cut cord cross sections approx. 1-1.5 mm. thick which were osmicated in a 1.0% OsO4/cacodylate buffer for 1.5 hrs. When I cut into the middle of the tissue block it was completely white.......the osmium did not penetrate beyond the surface of the tissue. I know that the heavily myelinated spinal cord can be a barrier to complete diffusion of osmium post fixation. These blocks will be sectioned for light microscopy stained with Toluidine Blue to access myelin fiber numbers, then potentially TEM examination.
Question: I have researched the literature and one paper suggested using 2.0% osmium and leaving the spinal cord blocks in overnight. Another suggested using potassium ferrocyanide with the osmium. One of our university EM experienced researchers suggested warming the osmium to 37 C. Any help would be greatly appreciated as I need an answer quickly.
Karen Bentley
Karen L. Bentley, M.S. Technical Director Electron Microscope Research Core University of Rochester Medical Center 575 Elmwood Avenue, Box 626 Rochester, NY 14642 585-275-1954
==============================Original Headers============================== 4, 26 -- From Karen_Bentley-at-URMC.Rochester.edu Thu May 24 09:32:43 2007 4, 26 -- Received: from w2k3ses2.urmc-sh.rochester.edu (ses02.urmc.rochester.edu [128.151.10.28]) 4, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4OEWhkW012749 4, 26 -- for {microscopy-at-microscopy.com} ; Thu, 24 May 2007 09:32:43 -0500 4, 26 -- Received: from mail pickup service by w2k3ses2.urmc-sh.rochester.edu with Microsoft SMTPSVC; 4, 26 -- Thu, 24 May 2007 10:32:42 -0400 4, 26 -- Received: from exsmtp02.urmc-sh.rochester.edu ([128.151.10.25]) by W2K3SES2.urmc-sh.rochester.edu; 4, 26 -- 24 May 2007 10:32:39 -0500 4, 26 -- Received: from e2k3ms2.urmc-sh.rochester.edu ([172.18.153.121]) by exsmtp02.urmc-sh.rochester.edu with Microsoft SMTPSVC(6.0.3790.1830); 4, 26 -- Thu, 24 May 2007 10:32:39 -0400 4, 26 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 4, 26 -- Content-class: urn:content-classes:message 4, 26 -- MIME-Version: 1.0 4, 26 -- Content-Type: text/plain; 4, 26 -- charset="us-ascii" 4, 26 -- Subject: Myelin Osmication Problem 4, 26 -- Date: Thu, 24 May 2007 10:32:13 -0400 4, 26 -- Message-ID: {26A221DAB4C7EE45A2A73DD9B33A1FA501597664-at-e2k3ms2.urmc-sh.rochester.edu} 4, 26 -- Thread-Topic: Myelin Osmication Problem 4, 26 -- thread-index: AceeEFHITwdKxwoNQ5uZrTf7mP3Ngw== 4, 26 -- From: "Bentley, Karen" {Karen_Bentley-at-URMC.Rochester.edu} 4, 26 -- To: {microscopy-at-microscopy.com} 4, 26 -- X-OriginalArrivalTime: 24 May 2007 14:32:39.0789 (UTC) FILETIME=[61A001D0:01C79E10] 4, 26 -- X-PRIVAWALL-ID: 00145e0b7c32_236c041 4, 26 -- Content-Transfer-Encoding: 8bit 4, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4OEWhkW012749 ==============================End of - Headers==============================
Is is possible for you to cut thinner cross-sections? I am working with a project that hands over to me fixed (similar fix to yours) rat spinal and ganglia sections around 40 micron. They are fully osmicated, black all the way through. I think they use a vibratome to cut those sections. Gib -- Gilbert (Gib) Ahlstrand, Electron Microscopist, Imaging Center, University of Minnesota 123 Snyder Hall St. Paul, MN 55108 http://www.cbs.umn.edu/ic
Karen_Bentley-at-URMC.Rochester.edu wrote: } } Hello Fellow Listers: } } I have an issue regarding perfusion fixed (glut & paraform. in } cacodylate buffer) adult rat spinal cord. } } I have cut cord cross sections approx. 1-1.5 mm. thick which were } osmicated in a 1.0% OsO4/cacodylate buffer for 1.5 hrs. When I cut into } the middle of the tissue block it was completely white.......the osmium } did not penetrate beyond the surface of the tissue. I know that the } heavily myelinated spinal cord can be a barrier to complete diffusion of } osmium post fixation. These blocks will be sectioned for light } microscopy stained with Toluidine Blue to access myelin fiber numbers, } then potentially TEM examination. } } Question: I have researched the literature and one paper suggested } using 2.0% osmium and leaving the spinal cord blocks in overnight. } Another suggested using potassium ferrocyanide with the osmium. One of } our university EM experienced researchers suggested warming the osmium } to 37 C. Any help would be greatly appreciated as I need an answer } quickly. } } Karen Bentley } } Karen L. Bentley, M.S. } Technical Director } Electron Microscope Research Core } University of Rochester Medical Center } 575 Elmwood Avenue, Box 626 } Rochester, NY 14642 } 585-275-1954
==============================Original Headers============================== 4, 21 -- From ahlst007-at-umn.edu Thu May 24 09:48:53 2007 4, 21 -- Received: from mta-w2.tc.umn.edu (mta-w2.tc.umn.edu [134.84.119.6]) 4, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4OEmrca024457 4, 21 -- for {Microscopy-at-Microscopy.com} ; Thu, 24 May 2007 09:48:53 -0500 4, 21 -- Received: from [160.94.39.28] (x160-94-39-28.cbs.umn.edu [160.94.39.28]) 4, 21 -- by mta-w2.tc.umn.edu (UMN smtpd) with ESMTP 4, 21 -- for {Microscopy-at-Microscopy.com} ; Thu, 24 May 2007 09:48:46 -0500 (CDT) 4, 21 -- X-Umn-Remote-Mta: [N] x160-94-39-28.cbs.umn.edu [160.94.39.28] #+LO+TS+AU+HN 4, 21 -- Message-ID: {4655A52B.4050301-at-umn.edu} 4, 21 -- Date: Thu, 24 May 2007 09:46:03 -0500 4, 21 -- From: Gib Ahlstrand {ahlst007-at-umn.edu} 4, 21 -- Reply-To: ahlst007-at-umn.edu 4, 21 -- Organization: Imaging Center UM 4, 21 -- User-Agent: Thunderbird 1.5.0.10 (Macintosh/20070221) 4, 21 -- MIME-Version: 1.0 4, 21 -- To: Microscopy-at-Microscopy.com 4, 21 -- Subject: Re: [Microscopy] Myelin Osmication Problem 4, 21 -- References: {200705241435.l4OEZm6p018374-at-ns.microscopy.com} 4, 21 -- In-Reply-To: {200705241435.l4OEZm6p018374-at-ns.microscopy.com} 4, 21 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 21 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Phil You might also want to check out Hitachi's Tabletop Microscope TM-1000 "Designed to fulfill the needs of applications that require more information than can be provided by a light microscope but doesn't require the expertise of a dedicated SEM operator" Don't own one, but have been impressed with the instrument at a couple of shows. Seems robust and reliable. Rick,
Richard Harris Manager - Imaging, Information and Data Systems Biotron Experimental Climate Change Research Facility University of Western Ontario, London Ontario, CANADA. N6A 5B7 Ph. 519-661-2111 ext. 86780 Fax 519-661-3935 e-mail rjharris-at-uwo.ca web: www.biotron.uwo.ca
-----Original Message----- X-from: Phil.Ahrenkiel-at-sdsmt.edu [mailto:Phil.Ahrenkiel-at-sdsmt.edu] Sent: Wednesday, May 23, 2007 3:53 PM To: rjharris-at-uwo.ca
Greetings-
Our nanoscience program received some information about the Phenom-Ed from FEI, a tablettop SEM designed for education. We are considering whether it would useful for students to perform quick analysis of nanomaterials. Sounds like the instrument is still in the prototype stage, but we are wondering if anyone has any impressions they could share on this. From the price we were quoted, it might actually be within our budget.
Some info on this product is available at: http://investor.fei.com/phoenix.zhtml?c=60978&p=irol-newsArticle&ID=9682 99&highlight=
Thanks in advance for your feedback. -Phil ------------------------------------------ Phil Ahrenkiel, Assistant Professor Nanoscience and Nanoengineering Ph.D. Program South Dakota School of Mines and Technology 501 East St. Joseph Street Rapid City, South Dakota 57701 Office: EEP 221 Phone: (605)394-5238, Fax: (605)394-2365 Email: Phil.Ahrenkiel-at-sdsmt.edu
==============================Original Headers============================== 7, 20 -- From Phil.Ahrenkiel-at-sdsmt.edu Wed May 23 14:49:01 2007 7, 20 -- Received: from owa.sdsmt.edu (owa.sdsmt.edu [151.159.3.7]) 7, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4NJn0OO007452 7, 20 -- for {Microscopy-at-microscopy.com} ; Wed, 23 May 2007 14:49:00 -0500 7, 20 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 7, 20 -- Content-class: urn:content-classes:message 7, 20 -- MIME-Version: 1.0 7, 20 -- Content-Type: text/plain; 7, 20 -- charset="us-ascii" 7, 20 -- Subject: Phenom-Ed SEM 7, 20 -- Date: Wed, 23 May 2007 13:48:59 -0600 7, 20 -- Message-ID: {6FD01225FACD9C4AA3870724330536F4065B5B9F-at-sdsmt-ex01.SDSMT.LOCAL} 7, 20 -- X-MS-Has-Attach: 7, 20 -- X-MS-TNEF-Correlator: 7, 20 -- Thread-Topic: Phenom-Ed SEM 7, 20 -- thread-index: Acedc2ffJOT9yU/VTH23gIJB3edmtA== 7, 20 -- From: "Ahrenkiel, Scott \(Phil\) P." {Phil.Ahrenkiel-at-sdsmt.edu} 7, 20 -- To: {Microscopy-at-microscopy.com} 7, 20 -- Content-Transfer-Encoding: 8bit 7, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4NJn0OO007452 ==============================End of - Headers==============================
==============================Original Headers============================== 15, 26 -- From rjharris-at-uwo.ca Thu May 24 10:10:51 2007 15, 26 -- Received: from uwo.ca (v320-146-lb.its.uwo.ca [129.100.74.146]) 15, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4OFApoX003926 15, 26 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 24 May 2007 10:10:51 -0500 15, 26 -- Received: from chico.mail.uwo.pri (whelan.mail.uwo.pri [172.29.32.40]) 15, 26 -- by chico.mail.uwo.pri 15, 26 -- (Sun Java System Messaging Server 6.2-6.03 (built May 18 2006)) 15, 26 -- with ESMTP id {0JIJ00CZ0WU3HS00-at-chico.mail.uwo.pri} for 15, 26 -- Microscopy-at-MSA.Microscopy.Com; Thu, 24 May 2007 11:10:51 -0400 (EDT) 15, 26 -- Received: from RICKNOTEBOOK ([129.100.68.106]) 15, 26 -- by chico.mail.uwo.pri (Sun Java System Messaging Server 6.2-6.03 (built May 18 15, 26 -- 2006)) with ESMTPS id {0JIJ0075JWU2K1Q1-at-chico.mail.uwo.pri} for 15, 26 -- Microscopy-at-MSA.Microscopy.Com; Thu, 24 May 2007 11:10:51 -0400 (EDT) 15, 26 -- Date: Thu, 24 May 2007 11:10:52 -0400 15, 26 -- From: Richard Harris {rjharris-at-uwo.ca} 15, 26 -- Subject: RE: [Microscopy] Phenom-Ed SEM 15, 26 -- In-reply-to: {200705231952.l4NJqtFP016518-at-ns.microscopy.com} 15, 26 -- To: Phil.Ahrenkiel-at-sdsmt.edu 15, 26 -- Cc: MSA Listserver {Microscopy-at-MSA.Microscopy.Com} 15, 26 -- Message-id: {0JIJ0075KWU2K1Q1-at-chico.mail.uwo.pri} 15, 26 -- MIME-version: 1.0 15, 26 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 15, 26 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 15, 26 -- Content-type: text/plain; charset=us-ascii 15, 26 -- Content-transfer-encoding: 7BIT 15, 26 -- Thread-index: Acedc/T3TM+Qy9DRR8O/l5gT3WEczwAoVJYg ==============================End of - Headers==============================
Osmium will penetrate 1 mm maximum but if both surfaces of a cross section of sp. cord are exposed you 'should' be OK with 2 exposed surfaces. Are the sections really 1-1.5 mm thick? Are you agitating the tissue? Is the osmium reasonably fresh? I don't see how 2% osmium will solve the problem and ferrocyanide won't help if the osmium does not arrive at the 'target'. Warming the osmium might help but I don't see it making a huge difference. I suggest fresh osmium, longer times in osmium, frequent (continuous?) agitation. If that fails try using a Vibratome to cut 0.5 -0.8 mm sections. If you are sectioning for TEM are you really going to cut all the way through a 1.5 mm slice? If not, make the sections thinner.
Geoff
Karen_Bentley-at-URMC.Rochester.edu wrote:
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-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
==============================Original Headers============================== 8, 35 -- From mcauliff-at-umdnj.edu Thu May 24 10:38:36 2007 8, 35 -- Received: from zix01.umdnj.edu (zix01.UMDNJ.EDU [130.219.34.124]) 8, 35 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4OFcZY0015872 8, 35 -- for {microscopy-at-msa.microscopy.com} ; Thu, 24 May 2007 10:38:35 -0500 8, 35 -- Received: from zix01.umdnj.edu (ZixVPM [127.0.0.1]) 8, 35 -- by Outbound.umdnj.edu (Proprietary) with ESMTP id 7191FA7B86 8, 35 -- for {microscopy-at-msa.microscopy.com} ; Thu, 24 May 2007 11:38:34 -0400 (EDT) 8, 35 -- Received: from umdnj.edu (imail.UMDNJ.EDU [130.219.34.129]) 8, 35 -- by zix01.umdnj.edu (Proprietary) with ESMTP id 62E40A7B41 8, 35 -- for {microscopy-at-msa.microscopy.com} ; Thu, 24 May 2007 11:38:33 -0400 (EDT) 8, 35 -- Received: from ([130.219.34.131]) 8, 35 -- by imail.umdnj.edu with ESMTP id KP-BRACD.169660679; 8, 35 -- Thu, 24 May 2007 11:38:01 -0400 8, 35 -- Received: from conversion-daemon.Polaris.umdnj.edu by Polaris.umdnj.edu 8, 35 -- (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 2004)) 8, 35 -- id {0JIJ00K01XZKMW-at-Polaris.umdnj.edu} (original mail from mcauliff-at-umdnj.edu) 8, 35 -- for microscopy-at-msa.microscopy.com; Thu, 24 May 2007 11:38:01 -0400 (EDT) 8, 35 -- Received: from [127.0.0.1] ([10.138.2.123]) 8, 35 -- by Polaris.umdnj.edu (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 8, 35 -- 2004)) with ESMTP id {0JIJ00E76Y3D9N-at-Polaris.umdnj.edu} ; Thu, 8, 35 -- 24 May 2007 11:38:01 -0400 (EDT) 8, 35 -- Date: Thu, 24 May 2007 11:38:01 -0400 8, 35 -- From: Geoff McAuliffe {mcauliff-at-umdnj.edu} 8, 35 -- Subject: Re: [Microscopy] Myelin Osmication Problem 8, 35 -- In-reply-to: {200705241433.l4OEXkQ3014514-at-ns.microscopy.com} 8, 35 -- To: Karen_Bentley-at-URMC.Rochester.edu, 8, 35 -- MicroscopyListserver {microscopy-at-msa.microscopy.com} 8, 35 -- Message-id: {4655B159.3070407-at-umdnj.edu} 8, 35 -- MIME-version: 1.0 8, 35 -- Content-type: text/plain; format=flowed; charset=ISO-8859-1 8, 35 -- Content-transfer-encoding: 7BIT 8, 35 -- X-Accept-Language: en-us, en 8, 35 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 8, 35 -- Gecko/20040804 Netscape/7.2 (ax) 8, 35 -- References: {200705241433.l4OEXkQ3014514-at-ns.microscopy.com} ==============================End of - Headers==============================
I got another telephone call at my lab yesterday from a solicitor. My phone number here is unlisted and does not show up on any caller id. I am getting 1 or 2 of these calls a week, too many to be random hits. I think some companies are harvesting my number from Listserv postings. Has anyone else had this experience?
Geoff
-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
==============================Original Headers============================== 6, 33 -- From mcauliff-at-umdnj.edu Thu May 24 10:56:31 2007 6, 33 -- Received: from zix03.umdnj.edu (zix03.UMDNJ.EDU [130.219.34.126]) 6, 33 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4OFuU5T027644 6, 33 -- for {microscopy-at-msa.microscopy.com} ; Thu, 24 May 2007 10:56:31 -0500 6, 33 -- Received: from zix03.umdnj.edu (ZixVPM [127.0.0.1]) 6, 33 -- by Outbound.umdnj.edu (Proprietary) with ESMTP id 048CA4BF73 6, 33 -- for {microscopy-at-msa.microscopy.com} ; Thu, 24 May 2007 11:56:30 -0400 (EDT) 6, 33 -- Received: from umdnj.edu (imail2.UMDNJ.EDU [130.219.34.139]) 6, 33 -- by zix03.umdnj.edu (Proprietary) with ESMTP id EAA034BFC4 6, 33 -- for {microscopy-at-msa.microscopy.com} ; Thu, 24 May 2007 11:45:51 -0400 (EDT) 6, 33 -- Received: from ([130.219.34.131]) 6, 33 -- by imail2.umdnj.edu with ESMTP id KP-BRCRF.102886870; 6, 33 -- Thu, 24 May 2007 11:45:19 -0400 6, 33 -- Received: from conversion-daemon.Polaris.umdnj.edu by Polaris.umdnj.edu 6, 33 -- (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 2004)) 6, 33 -- id {0JIJ00L01Y9LKS-at-Polaris.umdnj.edu} (original mail from mcauliff-at-umdnj.edu) 6, 33 -- for microscopy-at-msa.microscopy.com; Thu, 24 May 2007 11:45:19 -0400 (EDT) 6, 33 -- Received: from [127.0.0.1] ([10.138.2.123]) 6, 33 -- by Polaris.umdnj.edu (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 6, 33 -- 2004)) with ESMTP id {0JIJ00EFGYFI9N-at-Polaris.umdnj.edu} ; Thu, 6, 33 -- 24 May 2007 11:45:19 -0400 (EDT) 6, 33 -- Date: Thu, 24 May 2007 11:45:18 -0400 6, 33 -- From: Geoff McAuliffe {mcauliff-at-umdnj.edu} 6, 33 -- Subject: telephone solicitations 6, 33 -- To: MicroscopyListserver {microscopy-at-msa.microscopy.com} , 6, 33 -- Histonet {histonet-at-pathology.swmed.edu} 6, 33 -- Message-id: {4655B30E.6030502-at-umdnj.edu} 6, 33 -- MIME-version: 1.0 6, 33 -- Content-type: text/plain; format=flowed; charset=ISO-8859-1 6, 33 -- Content-transfer-encoding: 7BIT 6, 33 -- X-Accept-Language: en-us, en 6, 33 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 6, 33 -- Gecko/20040804 Netscape/7.2 (ax) ==============================End of - Headers==============================
Geoff, this is why I don't list my phone number in my Histonet posts any more. I was getting unsolicited phone calls.
Teri Johnson, HT(ASCP)QIHC Managing Director Histology Facility Stowers Institute for Medical Research 1000 E. 50th St. Kansas City, MO 64110
Dear Lists:
I got another telephone call at my lab yesterday from a solicitor. My phone number here is unlisted and does not show up on any caller id. I am getting 1 or 2 of these calls a week, too many to be random hits. I
think some companies are harvesting my number from Listserv postings. Has anyone else had this experience?
Geoff
-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
==============================Original Headers============================== 6, 33 -- From mcauliff-at-umdnj.edu Thu May 24 10:56:31 2007 6, 33 -- Received: from zix03.umdnj.edu (zix03.UMDNJ.EDU [130.219.34.126]) 6, 33 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4OFuU5T027644 6, 33 -- for {microscopy-at-msa.microscopy.com} ; Thu, 24 May 2007 10:56:31 -0500 6, 33 -- Received: from zix03.umdnj.edu (ZixVPM [127.0.0.1]) 6, 33 -- by Outbound.umdnj.edu (Proprietary) with ESMTP id 048CA4BF73 6, 33 -- for {microscopy-at-msa.microscopy.com} ; Thu, 24 May 2007 11:56:30 -0400 (EDT) 6, 33 -- Received: from umdnj.edu (imail2.UMDNJ.EDU [130.219.34.139]) 6, 33 -- by zix03.umdnj.edu (Proprietary) with ESMTP id EAA034BFC4 6, 33 -- for {microscopy-at-msa.microscopy.com} ; Thu, 24 May 2007 11:45:51 -0400 (EDT) 6, 33 -- Received: from ([130.219.34.131]) 6, 33 -- by imail2.umdnj.edu with ESMTP id KP-BRCRF.102886870; 6, 33 -- Thu, 24 May 2007 11:45:19 -0400 6, 33 -- Received: from conversion-daemon.Polaris.umdnj.edu by Polaris.umdnj.edu 6, 33 -- (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 2004)) 6, 33 -- id {0JIJ00L01Y9LKS-at-Polaris.umdnj.edu} (original mail from mcauliff-at-umdnj.edu) 6, 33 -- for microscopy-at-msa.microscopy.com; Thu, 24 May 2007 11:45:19 -0400 (EDT) 6, 33 -- Received: from [127.0.0.1] ([10.138.2.123]) 6, 33 -- by Polaris.umdnj.edu (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 6, 33 -- 2004)) with ESMTP id {0JIJ00EFGYFI9N-at-Polaris.umdnj.edu} ; Thu, 6, 33 -- 24 May 2007 11:45:19 -0400 (EDT) 6, 33 -- Date: Thu, 24 May 2007 11:45:18 -0400 6, 33 -- From: Geoff McAuliffe {mcauliff-at-umdnj.edu} 6, 33 -- Subject: telephone solicitations 6, 33 -- To: MicroscopyListserver {microscopy-at-msa.microscopy.com} , 6, 33 -- Histonet {histonet-at-pathology.swmed.edu} 6, 33 -- Message-id: {4655B30E.6030502-at-umdnj.edu} 6, 33 -- MIME-version: 1.0 6, 33 -- Content-type: text/plain; format=flowed; charset=ISO-8859-1 6, 33 -- Content-transfer-encoding: 7BIT 6, 33 -- X-Accept-Language: en-us, en 6, 33 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 6, 33 -- Gecko/20040804 Netscape/7.2 (ax) ==============================End of - Headers==============================
==============================Original Headers============================== 12, 21 -- From tjj-at-stowers-institute.org Thu May 24 11:10:33 2007 12, 21 -- Received: from stowers-institute.org (mail.stowers-institute.org [204.56.6.8]) 12, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4OGAXwI009119 12, 21 -- for {microscopy-at-msa.microscopy.com} ; Thu, 24 May 2007 11:10:33 -0500 12, 21 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 12, 21 -- Content-class: urn:content-classes:message 12, 21 -- MIME-Version: 1.0 12, 21 -- Content-Type: text/plain; 12, 21 -- charset="us-ascii" 12, 21 -- Subject: RE: [Microscopy] telephone solicitations 12, 21 -- Date: Thu, 24 May 2007 11:10:14 -0500 12, 21 -- Message-ID: {C28BAF593DC3314E9C0F3A50191C2E7807A3E0A1-at-EXCHKC03.stowers-institute.org} 12, 21 -- In-Reply-To: {200705241602.l4OG2Kws006559-at-ns.microscopy.com} 12, 21 -- X-MS-Has-Attach: 12, 21 -- X-MS-TNEF-Correlator: 12, 21 -- Thread-Topic: [Microscopy] telephone solicitations 12, 21 -- Thread-Index: AceeHPXk71s52vdfTpmbvuAsPCQbgQAAO87A 12, 21 -- From: "Johnson, Teri" {TJJ-at-stowers-institute.org} 12, 21 -- To: {microscopy-at-msa.microscopy.com} 12, 21 -- Content-Transfer-Encoding: 8bit 12, 21 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4OGAXwI009119 ==============================End of - Headers==============================
Geoff- I haven't been getting calls, but I do know that if I "Google" myself (don't we all do that sometimes?), I get pages of information, some of which is from my facilities' websites, some reference publications, but many hits are postings to this list. I am also frequently asked about my pricing for services, based on a price list that hasn't been on the facility's website in years, but persists out there on the www. We all must remember that there is no way to "erase" things once they hit cyberspace: a fact many new college grads are finding out when prospective employers find their embarrassing "My Space" postings. If these calls bother you, may I suggest that you create a alternate "signature" for your emails to the list, omitting your phone number. MSA members can find you from the member directory if we need to. That listing is password protected so outsiders can't get the info from there. Just remember, those of us who are frequent users of the listserver have put ourselves out in public through no fault of our wonderful Sysop, or anyone else. Its just the nature of the beast. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Weill-Cornell Medical College
On a similar note: I received a strange call from the publication Photonics Spectra two days ago. They read all my address info to me then asked me "What color are your eyes?". I told the woman that was a totally inappropriate question and she said they made them ask. I hung up - it was too weird.
Made me wonder if anyone else has had a bizarre exchange with them.
Beth
On May 24, 2007, at 11:57 AM, mcauliff-at-umdnj.edu wrote:
} } } } ---------------------------------------------------------------------- } ------ } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } Dear Lists: } } I got another telephone call at my lab yesterday from a solicitor. } My phone number here is unlisted and does not show up on any caller } id. } I am getting 1 or 2 of these calls a week, too many to be random } hits. I } think some companies are harvesting my number from Listserv postings. } Has anyone else had this experience? } } Geoff } } -- } -- } ********************************************** } Geoff McAuliffe, Ph.D. } Neuroscience and Cell Biology } Robert Wood Johnson Medical School } 675 Hoes Lane, Piscataway, NJ 08854 } voice: (732)-235-4583; fax: -4029 } mcauliff-at-umdnj.edu } ********************************************** } } } } ==============================Original } Headers============================== } 6, 33 -- From mcauliff-at-umdnj.edu Thu May 24 10:56:31 2007 } 6, 33 -- Received: from zix03.umdnj.edu (zix03.UMDNJ.EDU } [130.219.34.126]) } 6, 33 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP } id l4OFuU5T027644 } 6, 33 -- for {microscopy-at-msa.microscopy.com} ; Thu, 24 May 2007 } 10:56:31 -0500 } 6, 33 -- Received: from zix03.umdnj.edu (ZixVPM [127.0.0.1]) } 6, 33 -- by Outbound.umdnj.edu (Proprietary) with ESMTP id 048CA4BF73 } 6, 33 -- for {microscopy-at-msa.microscopy.com} ; Thu, 24 May 2007 } 11:56:30 -0400 (EDT) } 6, 33 -- Received: from umdnj.edu (imail2.UMDNJ.EDU [130.219.34.139]) } 6, 33 -- by zix03.umdnj.edu (Proprietary) with ESMTP id EAA034BFC4 } 6, 33 -- for {microscopy-at-msa.microscopy.com} ; Thu, 24 May 2007 } 11:45:51 -0400 (EDT) } 6, 33 -- Received: from ([130.219.34.131]) } 6, 33 -- by imail2.umdnj.edu with ESMTP id KP-BRCRF.102886870; } 6, 33 -- Thu, 24 May 2007 11:45:19 -0400 } 6, 33 -- Received: from conversion-daemon.Polaris.umdnj.edu by } Polaris.umdnj.edu } 6, 33 -- (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 } 2004)) } 6, 33 -- id {0JIJ00L01Y9LKS-at-Polaris.umdnj.edu} (original mail from } mcauliff-at-umdnj.edu) } 6, 33 -- for microscopy-at-msa.microscopy.com; Thu, 24 May 2007 } 11:45:19 -0400 (EDT) } 6, 33 -- Received: from [127.0.0.1] ([10.138.2.123]) } 6, 33 -- by Polaris.umdnj.edu (iPlanet Messaging Server 5.2 HotFix } 2.02 (built Oct 21 } 6, 33 -- 2004)) with ESMTP id {0JIJ00EFGYFI9N-at-Polaris.umdnj.edu} ; } Thu, } 6, 33 -- 24 May 2007 11:45:19 -0400 (EDT) } 6, 33 -- Date: Thu, 24 May 2007 11:45:18 -0400 } 6, 33 -- From: Geoff McAuliffe {mcauliff-at-umdnj.edu} } 6, 33 -- Subject: telephone solicitations } 6, 33 -- To: MicroscopyListserver {microscopy-at-msa.microscopy.com} , } 6, 33 -- Histonet {histonet-at-pathology.swmed.edu} } 6, 33 -- Message-id: {4655B30E.6030502-at-umdnj.edu} } 6, 33 -- MIME-version: 1.0 } 6, 33 -- Content-type: text/plain; format=flowed; charset=ISO-8859-1 } 6, 33 -- Content-transfer-encoding: 7BIT } 6, 33 -- X-Accept-Language: en-us, en } 6, 33 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en- } US; rv:1.7.2) } 6, 33 -- Gecko/20040804 Netscape/7.2 (ax) } ==============================End of - } Headers==============================
********************************************************************** Beth Richardson Electron Microscopy Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
http://www.plantbio.uga.edu/emlab/
Phone - (706) 542-1790 & FAX - (706) 542-1805
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) *******************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
************************************************************************ *** The Friends of the Marine Institute - Join Today! www.friendsofugami.org
==============================Original Headers============================== 14, 21 -- From beth-at-plantbio.uga.edu Thu May 24 11:54:43 2007 14, 21 -- Received: from dogwood.plantbio.uga.edu (dogwood.plantbio.uga.edu [128.192.26.2]) 14, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4OGshg1008379 14, 21 -- for {microscopy-at-microscopy.com} ; Thu, 24 May 2007 11:54:43 -0500 14, 21 -- Received: from [128.192.26.46] ([128.192.26.46]) 14, 21 -- (authenticated user beth-at-plantbio.uga.edu) 14, 21 -- by dogwood.plantbio.uga.edu 14, 21 -- (using TLSv1/SSLv3 with cipher AES128-SHA (128 bits)); 14, 21 -- Thu, 24 May 2007 12:54:39 -0400 14, 21 -- In-Reply-To: {200705241557.l4OFv6oQ028686-at-ns.microscopy.com} 14, 21 -- References: {200705241557.l4OFv6oQ028686-at-ns.microscopy.com} 14, 21 -- Mime-Version: 1.0 (Apple Message framework v752.3) 14, 21 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 14, 21 -- Message-Id: {890E2B0A-6BDF-409F-9D3A-933FAB0D99C4-at-plantbio.uga.edu} 14, 21 -- Cc: microscopy microscopy {microscopy-at-microscopy.com} 14, 21 -- Content-Transfer-Encoding: 7bit 14, 21 -- From: Beth Richardson {beth-at-plantbio.uga.edu} 14, 21 -- Subject: Photonics telephone solicitations 14, 21 -- Date: Thu, 24 May 2007 12:54:42 -0400 14, 21 -- To: mcauliff-at-umdnj.edu 14, 21 -- X-Mailer: Apple Mail (2.752.3) ==============================End of - Headers==============================
Dear Michael, I would check on that. I know that my Hitachi S-3000N, although it has no Polaroid camera, still bases its magnification on a 4" X 5" image printout. I always remember that a full-screen image is showing about twice the stated magnification. Regards,
-----Original Message----- X-from: michael-at-shaffer.net [mailto:michael-at-shaffer.net] Sent: May 24, 2007 4:44 AM To: mager-at-interchange.ubc.ca
Phil writes ...
} Our nanoscience program received some information about the } Phenom-Ed from FEI, a tablettop SEM designed for education. } We are considering whether it would useful for students to } perform quick analysis of nanomaterials. ...
Regarding "nanomaterials", keep in mind that magnifications are not what they used to be. For example, it used to be that a SEM's mag number was based on a 4x5 polaroid, whereas now the mag number is based on a 10" window on the computer display. The white paper describes the Phenom capable of mag=20,000x which, relative to SEMs we are used to, would equate to 10,000x (1 cm = 1,000 nM on a 4x5).
Not to take away from what should be a very valuable and interesting tool in a science program for many high schools and colleges, and I only comment relative to the term "nanomaterials".
cheerios :o) michael shaffer
SEM/MLA Research Coordinator {http://www.mun.ca/creait/maf/} Inco Innovation Centre Memorial University St. John's, Newfoundland
==============================Original Headers============================== 7, 20 -- From michael-at-shaffer.net Thu May 24 06:36:19 2007 7, 20 -- Received: from n007.sc1.he.tucows.com (smtpout1446.sc1.he.tucows.com [64.97.157.146]) 7, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4OBaJp4007503 7, 20 -- for {Microscopy-at-microscopy.com} ; Thu, 24 May 2007 06:36:19 -0500 7, 20 -- Received: from roamingwolf (134.153.130.141) by n007.sc1.he.tucows.com (7.2.069.1) (authenticated as michael-at-shaffer.net) 7, 20 -- id 4630CC6B004AED22 for Microscopy-at-microscopy.com; Thu, 24 May 2007 11:36:18 +0000 7, 20 -- From: "michael shaffer" {michael-at-shaffer.net} 7, 20 -- To: "MSA Microscopy list" {Microscopy-at-microscopy.com} 7, 20 -- References: {200705231949.l4NJnIjx008103-at-ns.microscopy.com} 7, 20 -- Subject: RE: [Microscopy] Phenom-Ed SEM 7, 20 -- Date: Thu, 24 May 2007 09:09:09 -0230 7, 20 -- Message-ID: {000f01c79df8$2569cf10$8d829986-at-CREAIT.MUN.CA} 7, 20 -- MIME-Version: 1.0 7, 20 -- Content-Type: text/plain; 7, 20 -- charset="US-ASCII" 7, 20 -- Content-Transfer-Encoding: 7bit 7, 20 -- X-Mailer: Microsoft Office Outlook 11 7, 20 -- thread-index: AcedcuSS+ggTxhopQim0x4OAZEW60QAgrMaw 7, 20 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 7, 20 -- In-Reply-To: {200705231949.l4NJnIjx008103-at-ns.microscopy.com} ==============================End of - Headers==============================
==============================Original Headers============================== 16, 35 -- From mager-at-interchange.ubc.ca Thu May 24 11:54:46 2007 16, 35 -- Received: from mr1.mail-relay.ubc.ca (mr1.mail-relay.ubc.ca [137.82.45.1]) 16, 35 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4OGsjcw008401 16, 35 -- for {microscopy-at-microscopy.com} ; Thu, 24 May 2007 11:54:45 -0500 16, 35 -- Received: from mr1.mail-relay.ubc.ca (localhost [127.0.0.1]) 16, 35 -- by localhost (Postfix) with SMTP id 0BB62ED0D 16, 35 -- for {microscopy-at-microscopy.com} ; Thu, 24 May 2007 09:54:45 -0700 (PDT) 16, 35 -- Received: from mta2.interchange.ubc.ca (mta2.interchange.ubc.ca [142.103.145.70]) 16, 35 -- by mr1.mail-relay.ubc.ca (Postfix) with ESMTP 16, 35 -- for {microscopy-at-microscopy.com} ; Thu, 24 May 2007 09:54:44 -0700 (PDT) 16, 35 -- Received: from Mary (desk.staff.mmat.ubc.ca [137.82.16.178]) 16, 35 -- by smtp.interchange.ubc.ca 16, 35 -- (iPlanet Messaging Server 5.2 HotFix 1.21 (built Sep 8 2003)) 16, 35 -- with ESMTPS id {0JIK006DS1N8AM-at-smtp.interchange.ubc.ca} for 16, 35 -- microscopy-at-microscopy.com; Thu, 24 May 2007 09:54:44 -0700 (PDT) 16, 35 -- Date: Thu, 24 May 2007 09:53:39 -0700 16, 35 -- From: Mary Mager {mager-at-interchange.ubc.ca} 16, 35 -- Subject: RE: [Microscopy] RE: Phenom-Ed SEM 16, 35 -- In-reply-to: {200705241143.l4OBhpBo016256-at-ns.microscopy.com} 16, 35 -- To: michael-at-shaffer.net 16, 35 -- Cc: "'Microscopy'" {microscopy-at-microscopy.com} 16, 35 -- Reply-to: mager-at-interchange.ubc.ca 16, 35 -- Message-id: {0JIK006DT1N8AM-at-smtp.interchange.ubc.ca} 16, 35 -- Organization: Materials Eng. UBC 16, 35 -- MIME-version: 1.0 16, 35 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 16, 35 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 16, 35 -- Content-type: text/plain; charset=us-ascii 16, 35 -- Content-transfer-encoding: 7bit 16, 35 -- Thread-index: Aced+M4vCM3FNqqLRvqJL2aHemtI+QAKtR/g 16, 35 -- X-UBC-Scanned: Sophos PureMessage 5.3.1.294258, Antispam-Engine: 2.5.0.283055, Antispam-Data: 2007.5.24.93834 16, 35 -- X-UBC-Relayed: Relayed through mail-relay.ubc.ca 16, 35 -- X-PerlMx-Spam: Probability=7%, Report=__C230066_P5 0, __CP_MEDIA_BODY 0, __CP_URI_IN_BODY 0, __CT 0, __CTE 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __HAS_X_MAILER 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0, __STOCK_PHRASE_24 0, __STOCK_PHRASE_7 0, __USER_AGENT_MS_GENERIC 0 16, 35 -- X-Spam-Level: 16, 35 -- X-Spam-Flag: No ==============================End of - Headers==============================
Although I no longer subscribe to a number of "freebie" magazines, this question is one that is very commonly asked. Other questions might include: place of birth, hair color, height, etc. I suppose it is some way to moitor if the solicitors are connecting with the clients. I have never had a followup call from a supervisor, however, verifying the data. } } On a similar note: } I received a strange call from the publication Photonics Spectra two } days ago. } They read all my address info to me then asked me "What color are } your eyes?". } I told the woman that was a totally inappropriate question and she } said they made them ask. } I hung up - it was too weird. } } Made me wonder if anyone else has had a bizarre exchange with them. } } Beth } } On May 24, 2007, at 11:57 AM, mcauliff-at-umdnj.edu wrote: } } } } } } } } } ----------------------------------------------------------------------
John J. Bozzola, Ph.D., Director Integrated Microscopy & Graphics Expertise (IMAGE) Southern Illinois University 750 Communications Drive - MC 4402 Carbondale, IL 62901 Telephone: 618-453-3730
I get telephone solicitations from mortgage brokers on a daily basis. I am on the no-call list at home, but I don't think this applies to a work phone. Sure is annoying.
John Mardinly Intel
This does not represent an opinion of Intel Corporation.
-----Original Message----- X-from: lcgould-at-med.cornell.edu [mailto:lcgould-at-med.cornell.edu] Sent: Thursday, May 24, 2007 9:16 AM To: Mardinly, John
Geoff- I haven't been getting calls, but I do know that if I "Google" myself (don't we all do that sometimes?), I get pages of information, some of which is from my facilities' websites, some reference publications, but many hits are postings to this list. I am also frequently asked about my pricing for services, based on a price list that hasn't been on the facility's website in years, but persists out there on the www. We all must remember that there is no way to "erase" things once they hit cyberspace: a fact many new college grads are finding out when prospective employers find their embarrassing "My Space" postings. If these calls bother you, may I suggest that you create a alternate "signature" for your emails to the list, omitting your phone number. MSA members can find you from the member directory if we need to. That listing is password protected so outsiders can't get the info from there. Just remember, those of us who are frequent users of the listserver have put ourselves out in public through no fault of our wonderful Sysop, or anyone else. Its just the nature of the beast. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Weill-Cornell Medical College
Not a Tardigrade. The features I'd really like to see are obscured, but 04 looks to show the prosoma of an eriophyid mite. The other images fit this, especially the setation and cuticular decorations. A good example is at http://www.insectimages.org/browse/detail.cfm?imgnum=1318030 (Not by me.) There are other good images on this site as well.
The hand-like feature is really nifty, but if it were claws of a tardigrade, they'd be on the ends of all limbs, and there would be 2 pairs of limbs at the posterior end. 04 looks to show
Phil
} Dear All, } see some new images at } www.elektronenmikroskopie.info/absinthe_animal/index.htm } } Thanks for all your emails helping me solve this riddle. :-)) } } } Kind regards, } Stefan } } } ---------------------------------------------------------------------------- } ----------------------------------------- } Stefan Diller - Scientific Photography } Arndtstrasse 22 } D - 97072 Wuerzburg Germany } ++49 - 931 - 7848700 Phone } ++49 - 931 - 7848701 Fax } ++49 - 175 - 717 70 51 Cell-Phone } } Websites: } www.stefan-diller.com } www.elektronenmikroskopie.info } www.assisi.de } Anfahrt: http://mail.map24.com/stefan.diller } ---------------------------------------------------------------------------- -- Philip Oshel Microscopy Facility Supervisor Biology Department 024C Brooks Hall Central Michigan University Mt. Pleasant, MI 48859
==============================Original Headers============================== 4, 22 -- From oshel1pe-at-cmich.edu Thu May 24 13:39:45 2007 4, 22 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4OIdiFu003100 4, 22 -- for {Microscopy-at-microscopy.com} ; Thu, 24 May 2007 13:39:45 -0500 4, 22 -- Received: from egatea.central.cmich.local ([141.209.15.74]) 4, 22 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id l4OJ2iqo001336 4, 22 -- for {Microscopy-at-microscopy.com} ; Thu, 24 May 2007 15:02:44 -0400 4, 22 -- Received: from [141.209.160.249] ([141.209.160.249]) by egatea.central.cmich.local with Microsoft SMTPSVC(6.0.3790.1830); 4, 22 -- Thu, 24 May 2007 14:39:41 -0400 4, 22 -- Mime-Version: 1.0 4, 22 -- Message-Id: {f06230903c27b892d035e-at-[141.209.160.249]} 4, 22 -- In-Reply-To: {200705241809.l4OI9Zle021790-at-ns.microscopy.com} 4, 22 -- References: {200705241809.l4OI9Zle021790-at-ns.microscopy.com} 4, 22 -- Date: Thu, 24 May 2007 14:39:39 -0400 4, 22 -- To: Microscopy-at-microscopy.com 4, 22 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 4, 22 -- Subject: Re: [Microscopy] Absinthe animal - follow-up... 4, 22 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 4, 22 -- X-OriginalArrivalTime: 24 May 2007 18:39:41.0980 (UTC) FILETIME=[E456DDC0:01C79E32] 4, 22 -- X-CanItPRO-Stream: default 4, 22 -- X-Spam-Score: -3.5 () INFO_TLD,L_EXCH_MF 4, 22 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
That's an excellent point about magnification. I hadn't been seriously shopping for an SEM in a while and just noticed that issue for the first time at MSA last year. Our two older scopes are both setup to record the magnification as for a 120-mm Polaroid print. I noticed that features in 50kx images in the new scopes were not as big as they were in our scopes. It was explained that magnification was now calculated according to displayed size which was seldom only 120 mm across.
When did the new rule take effect? I have long been accustomed to interpreting SEM images as 120mm across as for the old Polaroid film. It didn't matter that the image was displayed on a CRT twice as large. I just assumed magnification was calculated according to the print. Maybe it phased in as Polaroids phased out.
Warren ________________________________________ X-from: michael-at-shaffer.net [mailto:michael-at-shaffer.net] Sent: Thu 5/24/2007 6:37 AM To: wesaia-at-iastate.edu
Hi Warren,
I don't think that there was a "rule" that changed. As long as the first document from a microscope was relatively inflexible (a negative or Polaroid), it was quite easy to go back and get good numbers with information like "mag: 10,000x". This already changed when people started to use computers to look at images. If you reduce the image size by a factor of 2 (to insert in a publication, for example), you have already gone from 10,000x to 5,000x. Now, with digital acquisition and the flexibility to acquire a certain number of "pixels", the "magnification" has become a dangerous concept. If you received an image in the mail and it says "10,000x", you would have to know it's entire history to judge the validity of the magnification and make measurements.
A better number is probably one that does not assume a certain size of the final image to specify dimensions. We used to specify the "full width" of the image in microns or nm, today with the digital images, it is better to specify the dimension of a single pixel. That can then be used to directly measure or to display a scale bar, or even to calcualte the real magnification on paper.
The advantage of using magnification is that it immediately gives you an idea of size. If you look at something that was acquired at 100,000x, you know that it must be small.
Michael Bode, Ph.D.
General Manager
OLYMPUS SOFT IMAGING SOLUTIONS 12596 West Bayaud Ave #300 Lakewood, CO 80228 USA Tel.: +1 (303) 234-9270 Fax.: +1 (303) 234-9271 E-mail: Mike.Bode-at-olympus-sis.com www.olympus-sis.com
-----Original Message----- X-from: wesaia-at-iastate.edu [mailto:wesaia-at-iastate.edu] Sent: Thursday, May 24, 2007 15:23 To: Mike Bode
That's an excellent point about magnification. I hadn't been seriously shopping for an SEM in a while and just noticed that issue for the first time at MSA last year. Our two older scopes are both setup to record the magnification as for a 120-mm Polaroid print. I noticed that features in 50kx images in the new scopes were not as big as they were in our scopes. It was explained that magnification was now calculated according to displayed size which was seldom only 120 mm across.
When did the new rule take effect? I have long been accustomed to interpreting SEM images as 120mm across as for the old Polaroid film. It didn't matter that the image was displayed on a CRT twice as large. I just assumed magnification was calculated according to the print. Maybe it phased in as Polaroids phased out.
Warren ________________________________________ X-from: michael-at-shaffer.net [mailto:michael-at-shaffer.net] Sent: Thu 5/24/2007 6:37 AM To: wesaia-at-iastate.edu
Magnification, as you have indicated, can be a dangerous concept especially when one deals with digital images (in TEM, SEM, and LM). In our beginning microscopy classes, we try to emphasize that the micrometer bar is what is to be displayed on the image, never the magnifaction. However, as a relative means of orientation, one could state in the legend that the original image was examined at a instrumental mangification of 10,000x, etc.
JB
} Hi Warren, } } I don't think that there was a "rule" that } changed. As long as the first document from a } microscope was relatively inflexible (a negative } or Polaroid), it was quite easy to go back and } get good numbers with information like "mag: } 10,000x". This already changed when people } started to use computers to look at images. If } you reduce the image size by a factor of 2 (to } insert in a publication, for example), you have } already gone from 10,000x to 5,000x. Now, with } digital acquisition and the flexibility to } acquire a certain number of "pixels", the } "magnification" has become a dangerous concept. } If you received an image in the mail and it says } "10,000x", you would have to know it's entire } history to judge the validity of the } magnification and make measurements. } } A better number is probably one that does not } assume a certain size of the final image to } specify dimensions. We used to specify the "full } width" of the image in microns or nm, today with } the digital images, it is better to specify the } dimension of a single pixel. That can then be } used to directly measure or to display a scale } bar, or even to calcualte the real magnification } on paper. } } The advantage of using magnification is that it } immediately gives you an idea of size. If you } look at something that was acquired at 100,000x, } you know that it must be small. } } } Michael Bode, Ph.D. } } General Manager } } OLYMPUS SOFT IMAGING SOLUTIONS } 12596 West Bayaud Ave #300 } Lakewood, CO 80228 } USA } Tel.: +1 (303) 234-9270 } Fax.: +1 (303) 234-9271 } E-mail: Mike.Bode-at-olympus-sis.com } www.olympus-sis.com } } -----Original Message----- } X-from: wesaia-at-iastate.edu [mailto:wesaia-at-iastate.edu] } Sent: Thursday, May 24, 2007 15:23 } To: Mike Bode } Subject: [Microscopy] Phenom-Ed SEM } } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The } Microscopy Society of America To } Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver
John J. Bozzola, Ph.D., Director Integrated Microscopy & Graphics Expertise (IMAGE) Southern Illinois University 750 Communications Drive - MC 4402 Carbondale, IL 62901 Telephone: 618-453-3730
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (vlk7-at-alfred.edu) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, May 24, 2007 at 09:18:07 ---------------------------------------------------------------------------
Email: vlk7-at-alfred.edu Name: Victoria Knox
Organization: Alfred University
Education: Undergraduate College
Location: Alfred, NY USA
Question: I've been working with a program called Image-Pro plus to measure the diameter of micron-sized fibers from SEM micrographs. I need some assistance in how the program is measureing these fibers because I am getting inaccurate data that is off by at least 25% each time. I have tried to get the data using different measurements but it is still off. Any help you can provide would be greatly appreciated.
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.com/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both kay-at-natural-immunogenics.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: kay-at-natural-immunogenics.com Name: Kay Mitchen
Title-Subject: [Filtered] TEM PHILLIPS EM400
Question: We are looking for training of 2 additional operators and maintenance of a TEM reference Phillips EM 400
Please contact me urgently - feel free to call at 954/979-0885
I found your message particularly interesting since you thought that the results deviated from an already known value by 25%.
As you seem to have found out rather quickly, image processing packages rarely if ever divulge the methods they employ to obtain a result.
Fibers can be difficult to work with unless some assumptions are made about the shape of the fiber. A fiber with an elliptical cross section can appear circular, and a circular cross section appears to be elliptical in all but one possible intersection. Non-convex fibers can be a real dilemma.
Suppose that you could measure the diameters accurately - right to the last possible digit of precision. If the fibers are slightly different in size, then you see a range of sizes. If you happen to look at only a few fibers or possibly a set of fibers with a particular diameter, then you might be led to think that the measurements are off.
The problem you are investigating may not be as simple or as knowable as you might have hoped. So let's see what you are doing and maybe collectively we can assist you in finding out what is happening.
Cheers ----- Original Message ----- X-from: {vlk7-at-alfred.edu} To: {rboehrin-at-vt.edu} Sent: Thursday, May 24, 2007 7:14 PM
I've just read a paper where the author talks about block staining with uranyl acetate and lead citrate. UAc yes, but Pbcitrate? Has anyone heard of this? Does it work? The pictures in the paper were very nice; good contrast and detail. With the recent talk about general lack of contrast in specimens these days (I quite agree on that; I find contrast poorer now than in the past), could this be a new method?
All opinions please!
Cheers,
Diana
==============================Original Headers============================== 4, 18 -- From dianavd-at-eye.usyd.edu.au Thu May 24 20:17:20 2007 4, 18 -- Received: from galen.med.usyd.edu.au (nugalen.med.usyd.edu.au [129.78.36.39]) 4, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4P1HJEH027003 4, 18 -- for {Microscopy-at-microscopy.com} ; Thu, 24 May 2007 20:17:19 -0500 4, 18 -- Received: from [129.78.203.144] (helo=[129.78.203.144]) 4, 18 -- by galen.med.usyd.edu.au with esmtp (Exim 4.43) 4, 18 -- id 1HrOQe-0000Y4-Js 4, 18 -- for Microscopy-at-microscopy.com; Fri, 25 May 2007 11:17:16 +1000 4, 18 -- Mime-Version: 1.0 (Apple Message framework v752.2) 4, 18 -- Content-Transfer-Encoding: 7bit 4, 18 -- Message-Id: {88725CDF-65F1-43A6-ACCA-234E4E16B9AC-at-eye.usyd.edu.au} 4, 18 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 4, 18 -- To: Microscopy {Microscopy-at-microscopy.com} 4, 18 -- From: Diana van Driel {dianavd-at-eye.usyd.edu.au} 4, 18 -- Subject: lead citrate and block staining 4, 18 -- Date: Fri, 25 May 2007 11:17:10 +1000 4, 18 -- X-Mailer: Apple Mail (2.752.2) 4, 18 -- X-Spam-Score: -4.4 (----) ==============================End of - Headers==============================
Hi Diana I use Pb in block and not on the grid. Works great. NO lead pepper. See my article in Microscopy Today, January 2007. David
_____________________
David Elliott Ph.D. Assistant Professor - Department of Cell Biology and Anatomy Director, Research Microscopy Core Service University of Arizona College of Medicine PO Box 245004 Tucson, AZ 85724
Voice: 520-626-7870 Fax: 520-626-2097
On May 24, 2007, at 6:20 PM, dianavd-at-eye.usyd.edu.au wrote:
} } } } ---------------------------------------------------------------------- } ------ } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } I've just read a paper where the author talks about block staining } with uranyl acetate and lead citrate. UAc yes, but Pbcitrate? Has } anyone heard of this? Does it work? The pictures in the paper were } very nice; good contrast and detail. With the recent talk about } general lack of contrast in specimens these days (I quite agree on } that; I find contrast poorer now than in the past), could this be a } new method? } } All opinions please! } } Cheers, } } Diana } } ==============================Original } Headers============================== } 4, 18 -- From dianavd-at-eye.usyd.edu.au Thu May 24 20:17:20 2007 } 4, 18 -- Received: from galen.med.usyd.edu.au } (nugalen.med.usyd.edu.au [129.78.36.39]) } 4, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP } id l4P1HJEH027003 } 4, 18 -- for {Microscopy-at-microscopy.com} ; Thu, 24 May 2007 } 20:17:19 -0500 } 4, 18 -- Received: from [129.78.203.144] (helo=[129.78.203.144]) } 4, 18 -- by galen.med.usyd.edu.au with esmtp (Exim 4.43) } 4, 18 -- id 1HrOQe-0000Y4-Js } 4, 18 -- for Microscopy-at-microscopy.com; Fri, 25 May 2007 11:17:16 } +1000 } 4, 18 -- Mime-Version: 1.0 (Apple Message framework v752.2) } 4, 18 -- Content-Transfer-Encoding: 7bit } 4, 18 -- Message-Id: {88725CDF-65F1-43A6- } ACCA-234E4E16B9AC-at-eye.usyd.edu.au} } 4, 18 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; } format=flowed } 4, 18 -- To: Microscopy {Microscopy-at-microscopy.com} } 4, 18 -- From: Diana van Driel {dianavd-at-eye.usyd.edu.au} } 4, 18 -- Subject: lead citrate and block staining } 4, 18 -- Date: Fri, 25 May 2007 11:17:10 +1000 } 4, 18 -- X-Mailer: Apple Mail (2.752.2) } 4, 18 -- X-Spam-Score: -4.4 (----) } ==============================End of - } Headers==============================
==============================Original Headers============================== 9, 23 -- From Elliott-at-arizona.edu Thu May 24 21:13:19 2007 9, 23 -- Received: from smtpgate.email.arizona.edu (frodo.email.arizona.edu [128.196.132.210]) 9, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4P2DJI9006925 9, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 24 May 2007 21:13:19 -0500 9, 23 -- Received: from frodos_amavis (amavis5.email.arizona.edu [10.0.0.208]) 9, 23 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id 3C8D937822; 9, 23 -- Thu, 24 May 2007 19:13:19 -0700 (MST) 9, 23 -- Received: from [192.168.0.30] (doctorelliott.us [70.57.226.104]) 9, 23 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id 11E4F37818; 9, 23 -- Thu, 24 May 2007 19:13:14 -0700 (MST) 9, 23 -- Mime-Version: 1.0 (Apple Message framework v752.2) 9, 23 -- In-Reply-To: {200705250120.l4P1KbGG032083-at-ns.microscopy.com} 9, 23 -- References: {200705250120.l4P1KbGG032083-at-ns.microscopy.com} 9, 23 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 9, 23 -- Message-Id: {94D92403-DE8D-4FF1-B2D7-4BE2158957CD-at-arizona.edu} 9, 23 -- Content-Transfer-Encoding: 7bit 9, 23 -- From: David Elliott {Elliott-at-arizona.edu} 9, 23 -- Subject: Re: [Microscopy] lead citrate and block staining 9, 23 -- Date: Thu, 24 May 2007 19:13:13 -0700 9, 23 -- To: dianavd-at-eye.usyd.edu.au, 9, 23 -- Microscopy ListServer {Microscopy-at-MSA.Microscopy.Com} 9, 23 -- X-Mailer: Apple Mail (2.752.2) 9, 23 -- X-Virus-Scanned: amavisd-new at email.arizona.edu ==============================End of - Headers==============================
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Question: I am looking for a an EM service technician. If you know of an independent EM service technician for a JEOL electron microscope who services EMs in California please contact me.
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Title-Subject: [Filtered] Re: [Microscopy] AskAMicroscopist: help with Image-Pro plus
Question: Victoria,
We use Image Pro Plus for measurements too. Feel free to send me an e-mail off line with a reference picture, and I can try to help you. Between the 6 of us that use it here, we could figure it out. It could be the aspect ratio for pixels in your calibration set-up, so that a fiber not parallel wth the x-axis of the picture would have an incorrect measurement. it could also be that you are transforming the picture (different size or resolution of the image) which is different from your calibration.
The problem with your beast is that I cannot recognize feet. This suggests me that it may be not be an adult but a larva. In thise case "Viel Glück" ;-)
LG Stephane
--- stefan.diller-at-t-online.de wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Dear All, } see some new images at } www.elektronenmikroskopie.info/absinthe_animal/index.htm } } Thanks for all your emails helping me solve this } riddle. :-)) } } } Kind regards, } Stefan } } } ---------------------------------------------------------------------------- } ----------------------------------------- } Stefan Diller - Scientific Photography } Arndtstrasse 22 } D - 97072 Wuerzburg Germany } ++49 - 931 - 7848700 Phone } ++49 - 931 - 7848701 Fax } ++49 - 175 - 717 70 51 Cell-Phone } } Websites: } www.stefan-diller.com } www.elektronenmikroskopie.info } www.assisi.de } Anfahrt: http://mail.map24.com/stefan.diller } ---------------------------------------------------------------------------- } ----------------------------------------- } } } } ==============================Original } Headers============================== } 9, 24 -- From stefan.diller-at-t-online.de Thu May 24 } 13:04:40 2007 } 9, 24 -- Received: from mailout05.sul.t-online.com } (mailout05.sul.t-online.com [194.25.134.82]) } 9, 24 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with ESMTP id } l4OI4er8012028 } 9, 24 -- for {microscopy-at-microscopy.com} ; Thu, 24 } May 2007 13:04:40 -0500 } 9, 24 -- Received: from fwd32.aul.t-online.de } 9, 24 -- by mailout05.sul.t-online.com with smtp } 9, 24 -- id 1HrHfz-0005QS-00; Thu, 24 May 2007 } 20:04:39 +0200 } 9, 24 -- Received: from dose } (rf-SewZVZedv9u20AhtJ7BKqksmL0udyUCAmN3rz-VLcEzVJDhv0cT-at-[91.10.224.114]) } by fwd32.sul.t-online.de } 9, 24 -- with smtp id 1HrHfh-03qgS00; Thu, 24 May } 2007 20:04:21 +0200 } 9, 24 -- Message-ID: } {00b401c79e2d$f439e3e0$0d02a8c0-at-dose} } 9, 24 -- From: "Stefan Diller" } {stefan.diller-at-t-online.de} } 9, 24 -- To: {microscopy-at-microscopy.com} } 9, 24 -- Subject: Absinthe animal - follow-up... } 9, 24 -- Date: Thu, 24 May 2007 20:04:17 +0200 } 9, 24 -- MIME-Version: 1.0 } 9, 24 -- Content-Type: text/plain; } 9, 24 -- charset="iso-8859-1" } 9, 24 -- Content-Transfer-Encoding: 7bit } 9, 24 -- X-Priority: 3 } 9, 24 -- X-MSMail-Priority: Normal } 9, 24 -- X-Mailer: Microsoft Outlook Express } 6.00.2800.1807 } 9, 24 -- X-MimeOLE: Produced By Microsoft MimeOLE } V6.00.2800.1896 } 9, 24 -- X-ID: } rf-SewZVZedv9u20AhtJ7BKqksmL0udyUCAmN3rz-VLcEzVJDhv0cT } 9, 24 -- X-TOI-MSGID: } 2bce43ae-753f-445a-9d5e-0b87319859a4 } ==============================End of - } Headers============================== }
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==============================Original Headers============================== 7, 21 -- From nizets2-at-yahoo.com Fri May 25 02:51:46 2007 7, 21 -- Received: from web37412.mail.mud.yahoo.com (web37412.mail.mud.yahoo.com [209.191.91.144]) 7, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l4P7pjp5014276 7, 21 -- for {microscopy-at-microscopy.com} ; Fri, 25 May 2007 02:51:46 -0500 7, 21 -- Received: (qmail 84207 invoked by uid 60001); 25 May 2007 07:51:45 -0000 7, 21 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 7, 21 -- s=s1024; d=yahoo.com; 7, 21 -- h=X-YMail-OSG:Received:Date:From:Subject:To:Cc:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding:Message-ID; 7, 21 -- b=a7ToXPP/Vn93BTTeD50isyBVokgwFNTRt1K8sBNRDOehqr4QqJ85IcFNBe8IAIR0Sn6B3QooOkdKSW3c2P3CT72BscLNoT3BErIwFUmDWrul8droB8k78Lv2M5NwZScJWj3BmQ06RiiamC3+WtMJ8ealKJpnThodBZMLzDc95k8=; 7, 21 -- X-YMail-OSG: g0um48QVM1kLsAjh3ZPj0H2f6msr36CpO6ApqCJuYszA_Gdz85RVicN2NZON13iyvh_yC.zPUdtRAdQDxIcfDQ9lW9nSsLscYXECcRNztRUwS71oRJEbI2YryWc2n2kSap3ou7krHP3MC5KN.hcVIMJ1_eTAEXnalB8ZeeWzMcrP 7, 21 -- Received: from [80.122.101.102] by web37412.mail.mud.yahoo.com via HTTP; Fri, 25 May 2007 00:51:45 PDT 7, 21 -- Date: Fri, 25 May 2007 00:51:45 -0700 (PDT) 7, 21 -- From: Stephane Nizet {nizets2-at-yahoo.com} 7, 21 -- Subject: Re: [Microscopy] Absinthe animal - follow-up... 7, 21 -- To: stefan.diller-at-t-online.de 7, 21 -- Cc: microscopy-at-microscopy.com 7, 21 -- In-Reply-To: {200705241810.l4OIA5lS023564-at-ns.microscopy.com} 7, 21 -- MIME-Version: 1.0 7, 21 -- Content-Type: text/plain; charset=iso-8859-1 7, 21 -- Content-Transfer-Encoding: 8bit 7, 21 -- Message-ID: {591203.82179.qm-at-web37412.mail.mud.yahoo.com} ==============================End of - Headers==============================
Sure, but if you see a scale bar next to your feature, you don't know it is small, you know its size directly! (and if the full scale bar is 50 nm, you know it is very small)
Stephane
} --- Mike.Bode-at-olympus-sis.com wrote: } } } } } The advantage of using magnification is that it } } immediately gives you an idea of size. If you look } } at something that was acquired at 100,000x, you } know } } that it must be small. } } } } } } Michael Bode, Ph.D. } } } } General Manager } } } } OLYMPUS SOFT IMAGING SOLUTIONS } } 12596 West Bayaud Ave #300 } } Lakewood, CO 80228 } } USA } } Tel.: +1 (303) 234-9270 } } Fax.: +1 (303) 234-9271 } } E-mail: Mike.Bode-at-olympus-sis.com } } www.olympus-sis.com } } } } } } } ____________________________________________________________________________________Be } a better Heartthrob. Get better relationship answers } from someone who knows. Yahoo! Answers - Check it } out. } http://answers.yahoo.com/dir/?link=list&sid=396545433 }
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==============================Original Headers============================== 5, 19 -- From nizets2-at-yahoo.com Fri May 25 02:59:42 2007 5, 19 -- Received: from web37405.mail.mud.yahoo.com (web37405.mail.mud.yahoo.com [209.191.91.137]) 5, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l4P7xeAc021325 5, 19 -- for {microscopy-at-microscopy.com} ; Fri, 25 May 2007 02:59:41 -0500 5, 19 -- Received: (qmail 76858 invoked by uid 60001); 25 May 2007 07:59:40 -0000 5, 19 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 5, 19 -- s=s1024; d=yahoo.com; 5, 19 -- h=X-YMail-OSG:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding:Message-ID; 5, 19 -- b=NmdhY3ijiTuM2oUXY8jqwxihSjzEP6uO5Rg1WVkR6nzt6HYVKpPCYOXvrMYNR9nRUoDLoDjS7gKzr77fjwKsTPDpV51vLdnCgKnpBzPuLdVCeXbUw/4HT6EvpVo/vmnFGDkrgvjoeIjUg6RmC+niHcscknK87lxU9QwxrtqPaGk=; 5, 19 -- X-YMail-OSG: YhjooJQVM1nUeI8bxd.W0gDzT2t8GgjHd345kCUuLcAupA8zaYlJDIx4qSHZ_KW_WHqGfAkiNwhXtcY0302iT2h_JPQ7Mqv5BbsOBrgPIIuBtwflP0Q- 5, 19 -- Received: from [80.122.101.102] by web37405.mail.mud.yahoo.com via HTTP; Fri, 25 May 2007 00:59:39 PDT 5, 19 -- Date: Fri, 25 May 2007 00:59:39 -0700 (PDT) 5, 19 -- From: Stephane Nizet {nizets2-at-yahoo.com} 5, 19 -- Subject: Re: [Microscopy] RE: Phenom-Ed SEM 5, 19 -- To: microscopy-at-microscopy.com 5, 19 -- MIME-Version: 1.0 5, 19 -- Content-Type: text/plain; charset=iso-8859-1 5, 19 -- Content-Transfer-Encoding: 8bit 5, 19 -- Message-ID: {985427.76539.qm-at-web37405.mail.mud.yahoo.com} ==============================End of - Headers==============================
Hi Karen! Long time, no chat! In my experience with rat sciatic nerve, my pieces were several millimeters long (oriented for x-section view), but tended to be less wide than spinal cord, so I typically had great osmium penetration. However, I noted that several samples that were very wide tended to have poor osmium penetration throughout, particularly if I sectioned too deeply.
Keeping that in mind, I suggest thinner tissue samples (no longer than 0.5mm), either by hand or vibratome. Secondly, minimize tissue loss during your initial block face trimming and obtain your sections ASAP.
Best regards,
Walter F. Bobrowski Sr. Scientist Pfizer Global R&D 2800 Plymouth Rd. Ann Arbor, MI 48105
TEL: 734-622-7814 FAX: 734-622-5718
"Keep away from small people who try to belittle your ambitions. Small people always do that, but the really great make you feel that you, too, can become great." Mark Twain 1835-1910, Humorist and Writer
-----Original Message----- X-from: Karen_Bentley-at-URMC.Rochester.edu [mailto:Karen_Bentley-at-URMC.Rochester.edu] Sent: Thursday, May 24, 2007 10:37 AM To: Bobrowski, Walter
Hello Fellow Listers:
I have an issue regarding perfusion fixed (glut & paraform. in cacodylate buffer) adult rat spinal cord.
I have cut cord cross sections approx. 1-1.5 mm. thick which were osmicated in a 1.0% OsO4/cacodylate buffer for 1.5 hrs. When I cut into the middle of the tissue block it was completely white.......the osmium did not penetrate beyond the surface of the tissue. I know that the heavily myelinated spinal cord can be a barrier to complete diffusion of osmium post fixation. These blocks will be sectioned for light microscopy stained with Toluidine Blue to access myelin fiber numbers, then potentially TEM examination.
Question: I have researched the literature and one paper suggested using 2.0% osmium and leaving the spinal cord blocks in overnight. Another suggested using potassium ferrocyanide with the osmium. One of our university EM experienced researchers suggested warming the osmium to 37 C. Any help would be greatly appreciated as I need an answer quickly.
Karen Bentley
Karen L. Bentley, M.S. Technical Director Electron Microscope Research Core University of Rochester Medical Center 575 Elmwood Avenue, Box 626 Rochester, NY 14642 585-275-1954
==============================Original Headers============================== 4, 26 -- From Karen_Bentley-at-URMC.Rochester.edu Thu May 24 09:32:43 2007 4, 26 -- Received: from w2k3ses2.urmc-sh.rochester.edu (ses02.urmc.rochester.edu [128.151.10.28]) 4, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4OEWhkW012749 4, 26 -- for {microscopy-at-microscopy.com} ; Thu, 24 May 2007 09:32:43 -0500 4, 26 -- Received: from mail pickup service by w2k3ses2.urmc-sh.rochester.edu with Microsoft SMTPSVC; 4, 26 -- Thu, 24 May 2007 10:32:42 -0400 4, 26 -- Received: from exsmtp02.urmc-sh.rochester.edu ([128.151.10.25]) by W2K3SES2.urmc-sh.rochester.edu; 4, 26 -- 24 May 2007 10:32:39 -0500 4, 26 -- Received: from e2k3ms2.urmc-sh.rochester.edu ([172.18.153.121]) by exsmtp02.urmc-sh.rochester.edu with Microsoft SMTPSVC(6.0.3790.1830); 4, 26 -- Thu, 24 May 2007 10:32:39 -0400 4, 26 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 4, 26 -- Content-class: urn:content-classes:message 4, 26 -- MIME-Version: 1.0 4, 26 -- Content-Type: text/plain; 4, 26 -- charset="us-ascii" 4, 26 -- Subject: Myelin Osmication Problem 4, 26 -- Date: Thu, 24 May 2007 10:32:13 -0400 4, 26 -- Message-ID: {26A221DAB4C7EE45A2A73DD9B33A1FA501597664-at-e2k3ms2.urmc-sh.rochester.edu} 4, 26 -- Thread-Topic: Myelin Osmication Problem 4, 26 -- thread-index: AceeEFHITwdKxwoNQ5uZrTf7mP3Ngw== 4, 26 -- From: "Bentley, Karen" {Karen_Bentley-at-URMC.Rochester.edu} 4, 26 -- To: {microscopy-at-microscopy.com} 4, 26 -- X-OriginalArrivalTime: 24 May 2007 14:32:39.0789 (UTC) FILETIME=[61A001D0:01C79E10] 4, 26 -- X-PRIVAWALL-ID: 00145e0b7c32_236c041 4, 26 -- Content-Transfer-Encoding: 8bit 4, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4OEWhkW012749 ==============================End of - Headers==============================
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==============================Original Headers============================== 19, 31 -- From Walter.Bobrowski-at-pfizer.com Fri May 25 07:34:51 2007 19, 31 -- Received: from gromsgoa01.pfizer.com (gromsgo.pfizer.com [148.168.224.84]) 19, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4PCYpKP011024 19, 31 -- for {microscopy-at-microscopy.com} ; Fri, 25 May 2007 07:34:51 -0500 19, 31 -- Received: from mopamrexc03.amer.pfizer.com (mopamrexc03.pfizer.com [170.116.30.69]) 19, 31 -- by gromsgoa01.pfizer.com (8.13.7/8.13.7) with ESMTP id l4PCYk9m017638; 19, 31 -- Fri, 25 May 2007 08:34:51 -0400 19, 31 -- Received: from groamrexc02.amer.pfizer.com ([172.30.8.169]) by mopamrexc03.amer.pfizer.com with Microsoft SMTPSVC(6.0.3790.1830); 19, 31 -- Fri, 25 May 2007 08:34:49 -0400 19, 31 -- Received: from anaamrexm01.amer.pfizer.com ([162.48.101.10]) by groamrexc02.amer.pfizer.com with Microsoft SMTPSVC(6.0.3790.1830); 19, 31 -- Fri, 25 May 2007 08:34:49 -0400 19, 31 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 19, 31 -- Content-class: urn:content-classes:message 19, 31 -- MIME-Version: 1.0 19, 31 -- Content-Type: text/plain; 19, 31 -- charset="us-ascii" 19, 31 -- Subject: RE: [Microscopy] Myelin Osmication Problem 19, 31 -- Date: Fri, 25 May 2007 08:34:47 -0400 19, 31 -- Message-ID: {CF89AF6ABD93B746A55BCCD9480A4B3D09284DBE-at-anaamrexm01.amer.pfizer.com} 19, 31 -- In-Reply-To: {200705241437.l4OEbMIn021510-at-ns.microscopy.com} 19, 31 -- X-MS-Has-Attach: 19, 31 -- X-MS-TNEF-Correlator: 19, 31 -- Thread-Topic: [Microscopy] Myelin Osmication Problem 19, 31 -- Thread-Index: AceeEQ0wEV1BhOxKQ/iyKvL3QW0hjgAtpQRg 19, 31 -- From: "Bobrowski, Walter" {Walter.Bobrowski-at-pfizer.com} 19, 31 -- To: {Karen_Bentley-at-URMC.Rochester.edu} , {microscopy-at-microscopy.com} 19, 31 -- X-OriginalArrivalTime: 25 May 2007 12:34:49.0326 (UTC) FILETIME=[15B6B8E0:01C79EC9] 19, 31 -- X-Proofpoint-Virus-Version: vendor=fsecure engine=4.65.5502:2.3.11,1.2.37,4.0.164 definitions=2007-05-25_02:2007-05-23,2007-05-25,2007-05-25 signatures=0 19, 31 -- X-Proofpoint-Spam-Reason: safe 19, 31 -- Content-Transfer-Encoding: 8bit 19, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4PCYpKP011024 ==============================End of - Headers==============================
No, you don't know the size directly. Not unless the measured object is exactly orthogonal to the projected sight line. Quick experiment: project a bright light on the wall, and hold up a pencil (or similar). Hold the pencil so its long axis is exactly at 90 degrees to the central axis of the light path. Pretend the enlarged size of the pencil shadow is magnification. Now, tip the pencil into different orientations. Notice how the size of the shadow changes? Projective geometry. Same thing with an SEM image. It's a 3D object whose image is projected onto a 2D plane. Unless the dimension desired is exactly orthongonal to the line of sight, then the projected size will be different than the true size by an unknown amount. This can be solved by tilting the image, taking stereopairs, and doing trigonometry. But it will still be wrong unless the instrument has been properly calibrated *and* unless the all of the effects the specimen processing steps have on the sample are exactly known. Mind, this all ignores the more subtle question of "where's the edge"? Given the signal that forms an SEM image come from an interaction volume, not a dimensionless point, then the edge of a feature is essentially smeared out over the size of that volume. This is not much of an issue, or even no issue, for most imaging, but at high magnifications and resolutions, it is.
Phil
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==============================Original Headers============================== 4, 23 -- From oshel1pe-at-cmich.edu Fri May 25 07:53:43 2007 4, 23 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 4, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4PCrh4u022930 4, 23 -- for {Microscopy-at-microscopy.com} ; Fri, 25 May 2007 07:53:43 -0500 4, 23 -- Received: from egatea.central.cmich.local ([141.209.15.74]) 4, 23 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id l4PDGgqo010007; 4, 23 -- Fri, 25 May 2007 09:16:42 -0400 4, 23 -- Received: from [141.209.160.249] ([141.209.160.249]) by egatea.central.cmich.local with Microsoft SMTPSVC(6.0.3790.1830); 4, 23 -- Fri, 25 May 2007 08:53:41 -0400 4, 23 -- Mime-Version: 1.0 4, 23 -- Message-Id: {f06230904c27c89891143-at-[141.209.160.249]} 4, 23 -- In-Reply-To: {200705250804.l4P849BF030109-at-ns.microscopy.com} 4, 23 -- References: {200705250804.l4P849BF030109-at-ns.microscopy.com} 4, 23 -- Date: Fri, 25 May 2007 08:53:38 -0400 4, 23 -- To: nizets2-at-yahoo.com 4, 23 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 4, 23 -- Subject: Re: [Microscopy] Phenom-Ed SEM 4, 23 -- Cc: Microscopy-at-microscopy.com 4, 23 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 4, 23 -- X-OriginalArrivalTime: 25 May 2007 12:53:41.0660 (UTC) FILETIME=[B8A32DC0:01C79ECB] 4, 23 -- X-CanItPRO-Stream: default 4, 23 -- X-Spam-Score: -4 () L_EXCH_MF 4, 23 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
I have a Polaron SC-502 sputter coater in the SEM lab. One of the Engineering faculty wants to coat samples with chromium, titanium and aluminum for an application other than SEM. Assuming I can get targets that fit the sputter coater, are there other issues that would prevent using these other metals?
Thank You
Dave Wilbur -- __________________________________ David J. Wilbur, Ph.D. Instrumentation Specialist Department of Chemistry Tufts University 62 Talbot Ave. Medford, MA 02155 voice: 617-627-2163 Fax: 617-627-3443 email: david.wilbur-at-tufts.edu __________________________________
==============================Original Headers============================== 3, 20 -- From david.wilbur-at-tufts.edu Fri May 25 08:04:11 2007 3, 20 -- Received: from beech.usg.tufts.edu (beech.usg.tufts.edu [130.64.1.92]) 3, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4PD4BCj002195 3, 20 -- for {Microscopy-at-Microscopy.Com} ; Fri, 25 May 2007 08:04:11 -0500 3, 20 -- Received: from dhcp-130-64-102-103.medford.tufts.edu ([130.64.102.103]:1120) 3, 20 -- by beech.usg.tufts.edu with esmtps (TLSv1:AES256-SHA:256) 3, 20 -- (Exim 4.60) 3, 20 -- (envelope-from {david.wilbur-at-tufts.edu} ) 3, 20 -- id 1HrZSl-0005gD-dz 3, 20 -- for Microscopy-at-Microscopy.Com; Fri, 25 May 2007 09:04:11 -0400 3, 20 -- Message-ID: {4656DECA.3060604-at-tufts.edu} 3, 20 -- Date: Fri, 25 May 2007 09:04:10 -0400 3, 20 -- From: David Wilbur {david.wilbur-at-tufts.edu} 3, 20 -- User-Agent: Thunderbird 2.0.0.0 (Windows/20070326) 3, 20 -- MIME-Version: 1.0 3, 20 -- To: Microscopy Listserver {Microscopy-at-Microscopy.Com} 3, 20 -- Subject: [Microscopy] Can I use my SEM sputter coater for metals other than 3, 20 -- gold? 3, 20 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 3, 20 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
We've been working rather intensively with PAX-it here for microscopy. I've done some image analysis work with Image-Pro, but I can't answer definitively about how they are doing their measurements. PAX-it's method for determining measurements is quite simple and easy to understand, however, and I'm sure that Chris Jahns up at MIS [mailto:chrisj-at-paxit.com] would be happy to take a look at your images and attempt to give you some dimensions.
PAX-it works quite simply by counting pixels and comparing them to a known reference that you have calibrated the program with at a specific "magnification" that you set, i.e., for light microscopy, you put a stage micrometer onto the microscope stage, choose your objective, focus, and capture the image. You tag that image with your parameters, and input those parameters into the database. Then you calibrate the measurement module by indicating precisely where your units of measurements are according to the micrometer. This gives PAX-it the number of pixels per micron, inch, etc. both horizontally and vertically, and the program will simply count pixels between designated measurement points and apply that specific conversion factor every time you present it with an image that you have designated as being captured with those particular parameters.
I'm not sure how people calibrate measurement distances with a SEM, but I do know that there are reference target particles used for calibrating particle size analyzers. If you could capture an image from silica beads, for example, with a known diameter of 10 microns, for example, you should be able to tell the accuracy of your system.
Robert Zonis Product Development Chemist, LMTC Sanford L.P. - A Newell Rubbermaid Company 3 Sharpie Way Shelbyville, TN 37160 Direct: +1 (931) 685-6635 Fax: +1 (931) 685-6613 robert.zonis-at-sanford.com www.papermate.com
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==============================Original Headers============================== 9, 35 -- From Robert.Zonis-at-sanford.com Fri May 25 08:09:46 2007 9, 35 -- Received: from mail42.messagelabs.com (mail42.messagelabs.com [216.82.244.163]) 9, 35 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l4PD9jH1013494 9, 35 -- for {Microscopy-at-microscopy.com} ; Fri, 25 May 2007 08:09:46 -0500 9, 35 -- X-VirusChecked: Checked 9, 35 -- X-Env-Sender: Robert.Zonis-at-sanford.com 9, 35 -- X-Msg-Ref: server-21.tower-42.messagelabs.com!1180098584!27843118!1 9, 35 -- X-StarScan-Version: 5.5.12.11; banners=sanford.com,-,- 9, 35 -- X-Originating-IP: [12.2.115.11] 9, 35 -- Received: (qmail 5218 invoked from network); 25 May 2007 13:09:44 -0000 9, 35 -- Received: from gnat.newellco.com (HELO nafepncsvpout1.nr.ad.newellco.com) (12.2.115.11) 9, 35 -- by server-21.tower-42.messagelabs.com with SMTP; 25 May 2007 13:09:44 -0000 9, 35 -- Received: from nafepncsefe02.nr.ad.newellco.com (unverified) by nafepncsvpout1.nr.ad.newellco.com 9, 35 -- (Content Technologies SMTPRS 4.3.12) with ESMTP id {T7fceb17b6f0a0599261794-at-nafepncsvpout1.nr.ad.newellco.com} ; 9, 35 -- Fri, 25 May 2007 08:09:44 -0500 9, 35 -- Received: from nashbsasebe01.nr.ad.newellco.com ([10.217.223.198]) by nafepncsefe02.nr.ad.newellco.com with Microsoft SMTPSVC(6.0.3790.1830); 9, 35 -- Fri, 25 May 2007 08:09:44 -0500 9, 35 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 9, 35 -- Content-class: urn:content-classes:message 9, 35 -- MIME-Version: 1.0 9, 35 -- Content-Type: text/plain; 9, 35 -- charset="iso-8859-1" 9, 35 -- Subject: Re:[Microscopy] AskAMicroscopist: help with Image-Pro plus 9, 35 -- Date: Fri, 25 May 2007 08:09:43 -0500 9, 35 -- Message-ID: {8809C39726DC5F43824A3D3BE5D0286A0C983D1C-at-nashbsasebe01.nr.ad.newellco.com} 9, 35 -- X-MS-Has-Attach: 9, 35 -- X-MS-TNEF-Correlator: 9, 35 -- Thread-Topic: Re:[Microscopy] AskAMicroscopist: help with Image-Pro plus 9, 35 -- Thread-Index: AceezfXAiEpwYzD4T+enY4Hvda6DpQ== 9, 35 -- From: "Zonis, Robert" {Robert.Zonis-at-sanford.com} 9, 35 -- To: {Microscopy-at-microscopy.com} 9, 35 -- Cc: {vlk7-at-alfred.edu} 9, 35 -- X-OriginalArrivalTime: 25 May 2007 13:09:44.0384 (UTC) FILETIME=[F6773400:01C79ECD] 9, 35 -- Content-Transfer-Encoding: 8bit 9, 35 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4PD9jH1013494 ==============================End of - Headers==============================
We use our Emitech coater to coat with a variety of metals, including chromium, platinum, tantalum, and others. You will have to check on sputtering currents and times and how they relate to coating thickness for whatever metal you're using, but I'm not personally aware of any reason you can't use the Polaron for a variety of metals.
My real reason for replying is to get one of the best-kept secrets in the World of Sputtering out there. You can get custom-made targets of virtually any metal from Refining Systems, Inc. in Las Vegas, NV. These targets can be made to your specified diameters, shapes and thicknesses, usually at substantially less cost than targets from coater manufacturers. The company is run by Mr. Abe Dayani, who has always answered the phone himself when I've called. Website is at http://www.refiningsystems.com/, phone is 702-368-0579.
I have heard that the purity of the metals used by the OEMs may be higher (like 99.998 vs. 99.99, for example), but I can say that we have never had any problem with the coatings from the RFI targets. It may be a factor for super-critical applications in engineering, so you may want to consider this.
I have no connection with RFI other than as a satisfied customer.
Cheers, Randy
Randy Tindall Senior EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount= Week&NavType=Both&Type=TimePlan
-----Original Message----- X-from: david.wilbur-at-tufts.edu [mailto:david.wilbur-at-tufts.edu] Sent: Friday, May 25, 2007 8:05 AM To: Tindall, Randy D.
I have a Polaron SC-502 sputter coater in the SEM lab. One of the Engineering faculty wants to coat samples with chromium, titanium and aluminum for an application other than SEM. Assuming I can get targets that fit the sputter coater, are there other issues that would prevent using these other metals?
Thank You
Dave Wilbur -- __________________________________ David J. Wilbur, Ph.D. Instrumentation Specialist Department of Chemistry Tufts University 62 Talbot Ave. Medford, MA 02155 voice: 617-627-2163 Fax: 617-627-3443 email: david.wilbur-at-tufts.edu __________________________________
==============================Original Headers============================== 3, 20 -- From david.wilbur-at-tufts.edu Fri May 25 08:04:11 2007 3, 20 -- Received: from beech.usg.tufts.edu (beech.usg.tufts.edu [130.64.1.92]) 3, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4PD4BCj002195 3, 20 -- for {Microscopy-at-Microscopy.Com} ; Fri, 25 May 2007 08:04:11 -0500 3, 20 -- Received: from dhcp-130-64-102-103.medford.tufts.edu ([130.64.102.103]:1120) 3, 20 -- by beech.usg.tufts.edu with esmtps (TLSv1:AES256-SHA:256) 3, 20 -- (Exim 4.60) 3, 20 -- (envelope-from {david.wilbur-at-tufts.edu} ) 3, 20 -- id 1HrZSl-0005gD-dz 3, 20 -- for Microscopy-at-Microscopy.Com; Fri, 25 May 2007 09:04:11 -0400 3, 20 -- Message-ID: {4656DECA.3060604-at-tufts.edu} 3, 20 -- Date: Fri, 25 May 2007 09:04:10 -0400 3, 20 -- From: David Wilbur {david.wilbur-at-tufts.edu} 3, 20 -- User-Agent: Thunderbird 2.0.0.0 (Windows/20070326) 3, 20 -- MIME-Version: 1.0 3, 20 -- To: Microscopy Listserver {Microscopy-at-Microscopy.Com} 3, 20 -- Subject: [Microscopy] Can I use my SEM sputter coater for metals other than 3, 20 -- gold? 3, 20 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 3, 20 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
==============================Original Headers============================== 15, 26 -- From TindallR-at-missouri.edu Fri May 25 08:26:24 2007 15, 26 -- Received: from um-tsmtpout1.um.umsystem.edu (um-tsmtpout1.um.umsystem.edu [209.106.228.23]) 15, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4PDQOW1025366 15, 26 -- for {microscopy-at-microscopy.com} ; Fri, 25 May 2007 08:26:24 -0500 15, 26 -- Received: from UM-XMAIL08.um.umsystem.edu ([209.106.228.34]) by um-tsmtpout1.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 15, 26 -- Fri, 25 May 2007 08:26:24 -0500 15, 26 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 15, 26 -- Content-class: urn:content-classes:message 15, 26 -- MIME-Version: 1.0 15, 26 -- Content-Type: text/plain; 15, 26 -- charset="us-ascii" 15, 26 -- Subject: RE: [Microscopy] Can I use my SEM sputter coater for metals other than 15, 26 -- Date: Fri, 25 May 2007 08:26:24 -0500 15, 26 -- Message-ID: {91108EF9255B394CBF8B7E3789814A4101C0BA63-at-UM-XMAIL08.um.umsystem.edu} 15, 26 -- In-Reply-To: {200705251305.l4PD59A2004131-at-ns.microscopy.com} 15, 26 -- X-MS-Has-Attach: 15, 26 -- X-MS-TNEF-Correlator: 15, 26 -- Thread-Topic: [Microscopy] Can I use my SEM sputter coater for metals other than 15, 26 -- Thread-Index: AceezVLU2lQSrKdNS8SkcTADx8rHwQAAJLwA 15, 26 -- References: {200705251305.l4PD59A2004131-at-ns.microscopy.com} 15, 26 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 15, 26 -- To: {david.wilbur-at-tufts.edu} 15, 26 -- Cc: {microscopy-at-microscopy.com} 15, 26 -- X-OriginalArrivalTime: 25 May 2007 13:26:24.0534 (UTC) FILETIME=[4A99FB60:01C79ED0] 15, 26 -- Content-Transfer-Encoding: 8bit 15, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4PDQOW1025366 ==============================End of - Headers==============================
I am not familiar with the specific sputter coater you are using. So here are some questions. Do you know if it is a straight DC sputter coater or magnitron sputter coater? What is the pumping system, just a mechanical pump?
Most sputter coaters for SEM sample prep only use a mechanical pump for vacuum generation. This means you are only going to about 10-3 torr of vacuum before backfilling and then coating. For most thin film depositions, this is quite poor vacuum. Most thin film sputter coaters will go to somewhere around 10-6 to 10-8 torr before backfilling for sputter deposition. They also usually have a way to clean the substrate prior to deposition, which your little sputter coater I imagine does not have. The amount of contamination you will have, both on the first coat and then when switching between targets will be much larger than usually considered acceptable for thin film deposition.
All that being said, I don't see any reason you can't use your machine to deposit the materials you are mentioning, but I do question how useful the resulting layered structure will end up being. I would have to know a lot more about what the desired end product is to give you my thoughts on that.
dj
On Fri, 25 May 2007, david.wilbur-at-tufts.edu wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } I have a Polaron SC-502 sputter coater in the SEM lab. One of the } Engineering faculty wants to coat samples with chromium, titanium and } aluminum for an application other than SEM. Assuming I can get targets } that fit the sputter coater, are there other issues that would prevent } using these other metals? } } Thank You } } Dave Wilbur } -- } __________________________________ } David J. Wilbur, Ph.D. } Instrumentation Specialist } Department of Chemistry } Tufts University } 62 Talbot Ave. } Medford, MA 02155 } voice: 617-627-2163 } Fax: 617-627-3443 } email: david.wilbur-at-tufts.edu } __________________________________ } } ==============================Original Headers============================== } 3, 20 -- From david.wilbur-at-tufts.edu Fri May 25 08:04:11 2007 } 3, 20 -- Received: from beech.usg.tufts.edu (beech.usg.tufts.edu [130.64.1.92]) } 3, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4PD4BCj002195 } 3, 20 -- for {Microscopy-at-Microscopy.Com} ; Fri, 25 May 2007 08:04:11 -0500 } 3, 20 -- Received: from dhcp-130-64-102-103.medford.tufts.edu ([130.64.102.103]:1120) } 3, 20 -- by beech.usg.tufts.edu with esmtps (TLSv1:AES256-SHA:256) } 3, 20 -- (Exim 4.60) } 3, 20 -- (envelope-from {david.wilbur-at-tufts.edu} ) } 3, 20 -- id 1HrZSl-0005gD-dz } 3, 20 -- for Microscopy-at-Microscopy.Com; Fri, 25 May 2007 09:04:11 -0400 } 3, 20 -- Message-ID: {4656DECA.3060604-at-tufts.edu} } 3, 20 -- Date: Fri, 25 May 2007 09:04:10 -0400 } 3, 20 -- From: David Wilbur {david.wilbur-at-tufts.edu} } 3, 20 -- User-Agent: Thunderbird 2.0.0.0 (Windows/20070326) } 3, 20 -- MIME-Version: 1.0 } 3, 20 -- To: Microscopy Listserver {Microscopy-at-Microscopy.Com} } 3, 20 -- Subject: [Microscopy] Can I use my SEM sputter coater for metals other than } 3, 20 -- gold? } 3, 20 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed } 3, 20 -- Content-Transfer-Encoding: 7bit } ==============================End of - Headers============================== }
==============================Original Headers============================== 8, 19 -- From dljones-at-bestweb.net Fri May 25 08:32:22 2007 8, 19 -- Received: from vms044pub.verizon.net (vms044pub.verizon.net [206.46.252.44]) 8, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4PDWM7C004042 8, 19 -- for {Microscopy-at-microscopy.com} ; Fri, 25 May 2007 08:32:22 -0500 8, 19 -- Received: from localhost ([71.247.102.125]) 8, 19 -- by vms044.mailsrvcs.net (Sun Java System Messaging Server 6.2-6.01 (built Apr 8, 19 -- 3 2006)) with ESMTPA id {0JIL00536MXI59AA-at-vms044.mailsrvcs.net} for 8, 19 -- Microscopy-at-microscopy.com; Fri, 25 May 2007 08:32:11 -0500 (CDT) 8, 19 -- Date: Fri, 25 May 2007 09:33:09 -0400 (Eastern Daylight Time) 8, 19 -- From: "David L. Jones" {dljones-at-bestweb.net} 8, 19 -- Subject: Re: [Microscopy] Can I use my SEM sputter coater for metals other than 8, 19 -- In-reply-to: {200705251307.l4PD7gEs008393-at-ns.microscopy.com} 8, 19 -- X-X-Sender: dljones-at-pop-croton.bestweb.net 8, 19 -- To: david.wilbur-at-tufts.edu 8, 19 -- Cc: Microscopy-at-microscopy.com 8, 19 -- Message-id: {Pine.WNT.4.64.0705250918100.3128-at-H-F1} 8, 19 -- MIME-version: 1.0 8, 19 -- Content-type: TEXT/PLAIN; charset=US-ASCII; format=flowed 8, 19 -- References: {200705251307.l4PD7gEs008393-at-ns.microscopy.com} ==============================End of - Headers==============================
I recommend that you download and install NIH Image (a.k.a. ImageJ). Using the same calibration image you used for ImagePro, does it give you the same numbers? If so, you might want to image a new calibration specimen. If it's still not what you expected, at least you'll know it isn't an error in the software. You might be seeing shrinkage due to specimen processing (critical point drying, etc.), exposure to vacuum, the 3d projection errors mentioned in another post, etc.
-----Original Message----- X-from: vlk7-at-alfred.edu [mailto:vlk7-at-alfred.edu] Sent: Thursday, May 24, 2007 5:15 PM To: Bowling, Andrew
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (vlk7-at-alfred.edu) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, May 24, 2007 at 09:18:07 ------------------------------------------------------------------------ ---
Email: vlk7-at-alfred.edu Name: Victoria Knox
Organization: Alfred University
Education: Undergraduate College
Location: Alfred, NY USA
Question: I've been working with a program called Image-Pro plus to measure the diameter of micron-sized fibers from SEM micrographs. I need some assistance in how the program is measureing these fibers because I am getting inaccurate data that is off by at least 25% each time. I have tried to get the data using different measurements but it is still off. Any help you can provide would be greatly appreciated.
Thanks Larry. Actually I concur with your experience with small animals (mice: 5-8 ml/min; rats: 10-20 ml/min). Interestingly I supported a colleague who needed monkey brain perfusion assistance and we simply followed a recommended protocol they had in hand (~300 ml/min). They were delighted with the histology results. Even I was shocked as the quality of preservation in spite of the high flow rate, as we typically recommend ~100 ml/min for large animals. I'm attempting to track down the reference.
In addition, Microscopy Today (May 2006) had a very relevant article on animal brain perfusion, advocating precise pressures, rather than flow rate, including a short bolus at 300 mm Hg, followed by 100 mm Hg.
Best regards, Walt
-----Original Message----- X-from: larry.ackerman-at-ucsf.edu [mailto:larry.ackerman-at-ucsf.edu] Sent: Wednesday, May 23, 2007 1:49 PM To: Bobrowski, Walter
Mary Ellen, The rate that Walter suggested seemed extremely high compared to my experience with smaller animals so I asked an anatomist with many decades of experience with monkeys. He has always used an initial flush of 500ml in 5 minutes time with buffer followed by 1.5 liters of fixative over 20 minutes of time for a typical 5kg animal. The appropriate size tubing and cannula in collaboration with the setting on
the pump will enable an excellent fixation. Walter's method may well provide good results. I would love to see some brain slices and TEM micrographs at 10,000X. I'm open to different methods.
Larry
Walter.Bobrowski-at-pfizer.com wrote: } ------------------------------------------------------------------------ ---- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ ---- } } 300 ml/min. Perfuse either through the carotid artery or left ventricle } (occlude descending aorta). } } Best regards, } Walter F. Bobrowski } Sr. Scientist } Pfizer Global R&D } 2800 Plymouth Rd. } Ann Arbor, MI 48105 } } TEL: 734-622-7814 } FAX: 734-622-5718 } } "Like the strength of a steel rod, the true character of a person can } only be known under extreme stress." - Leslie Fieger } } } } } -----Original Message----- } X-from: mpease-at-jhmi.edu [mailto:mpease-at-jhmi.edu] } Sent: Monday, May 21, 2007 7:25 PM } To: Bobrowski, Walter } Subject: [Microscopy] viaWWW: monkey perfusion methodology } } } } } ------------------------------------------------------------------------ } ---- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ------------------------------------------------------------------------ } ---- } } This Question/Comment was submitted to the Microscopy Listserver } using the WWW based Form at } http://www.microscopy.org/MicroscopyListserver/MLFormMail.html } ------------------------------------------------------------------------ } --- } Remember this posting is most likely not from a Subscriber, so when } replying } please copy both mpease-at-jhmi.edu as well as the MIcroscopy } Listserver } ------------------------------------------------------------------------ } --- } } Email: mpease-at-jhmi.edu } Name: Mary Ellen Pease } } Organization: Johns Hopkins Wilmer Eye Institute } } Title-Subject: [Filtered] monkey perfusion methodology } } Question: What is the recommended flow rate for perfusion of monkey } brain and eyes when using a pump rather than gravity feed of the } solutions? } } Thank you, } Mary Ellen } } ------------------------------------------------------------------------ } --- } } ==============================Original } Headers============================== } 7, 11 -- From zaluzec-at-microscopy.com Mon May 21 18:19:37 2007 } 7, 11 -- Received: from [206.69.208.22] (mac22.zaluzec.com } [206.69.208.22]) } 7, 11 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id l4LNJbJx007966 } 7, 11 -- for {microscopy-at-microscopy.com} ; Mon, 21 May 2007 } 18:19:37 -0500 } 7, 11 -- Mime-Version: 1.0 } 7, 11 -- Message-Id: {p06240806c277d979e099-at-[206.69.208.22]} } 7, 11 -- Date: Mon, 21 May 2007 18:19:36 -0500 } 7, 11 -- To: microscopy-at-microscopy.com } 7, 11 -- From: mpease-at-jhmi.edu (by way of MicroscopyListserver) } 7, 11 -- Subject: viaWWW: monkey perfusion methodology } 7, 11 -- Content-Type: text/plain; charset="us-ascii" } ==============================End of - } Headers============================== } } ---------------------------------------------------------------------- } LEGAL NOTICE } Unless expressly stated otherwise, this message is confidential and may be privileged. 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-- Larry Ackerman, Associate Specialist UCSF, Dept. of Anatomy, Rm S1347 513 Parnassus Ave., Box 0452 San Francisco, CA 94143
larry.ackerman-at-ucsf.edu
==============================Original Headers============================== 6, 28 -- From Larry.Ackerman-at-ucsf.edu Wed May 23 12:41:45 2007 6, 28 -- Received: from emfmcb02.ucsfmedicalcenter.org (emfmcb02.ucsfmedicalcenter.org [64.54.46.98]) 6, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4NHfijF002378 6, 28 -- for {Microscopy-at-microscopy.com} ; Wed, 23 May 2007 12:41:44 -0500 6, 28 -- Received: from 64.54.128.152 by emfmcb01.ucsfmedicalcenter.org with 6, 28 -- ESMTP (Tumbleweed Email Firewall SMTP Relay (Email Firewall v6.1.0)); 6, 28 -- Wed, 23 May 2007 10:53:00 -0700 6, 28 -- X-Server-Uuid: E2E48A14-EE5B-4280-A138-188440602EDD 6, 28 -- Received: from baluk.ucsf.edu ([128.218.123.88]) by exvs06.net.ucsf.edu 6, 28 -- with Microsoft SMTPSVC(6.0.3790.3959); Wed, 23 May 2007 10:41:19 -0700 6, 28 -- Message-ID: {46547CBF.8050702-at-ucsf.edu} 6, 28 -- Date: Wed, 23 May 2007 10:41:19 -0700 6, 28 -- From: "Larry Ackerman" {larry.ackerman-at-ucsf.edu} 6, 28 -- Reply-to: larry.ackerman-at-ucsf.edu 6, 28 -- Organization: UCSF, NeuroAnatomy 6, 28 -- User-Agent: Thunderbird 2.0.0.0 (Macintosh/20070326) 6, 28 -- MIME-Version: 1.0 6, 28 -- To: mpease-at-jhmi.edu, Microscopy-at-microscopy.com 6, 28 -- Subject: Re: [Microscopy] monkey perfusion methodology 6, 28 -- References: {200705221231.l4MCVVEO018334-at-ns.microscopy.com} 6, 28 -- In-Reply-To: {200705221231.l4MCVVEO018334-at-ns.microscopy.com} 6, 28 -- X-OriginalArrivalTime: 23 May 2007 17:41:19.0789 (UTC) 6, 28 -- FILETIME=[92752DD0:01C79D61] 6, 28 -- X-WSS-ID: 6A4AA0F00MC2362538-15-01 6, 28 -- Content-Type: text/plain; 6, 28 -- charset=iso-8859-1; 6, 28 -- format=flowed 6, 28 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
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==============================Original Headers============================== 17, 31 -- From Walter.Bobrowski-at-pfizer.com Fri May 25 08:52:51 2007 17, 31 -- Received: from gromsgoa01.pfizer.com (gromsgo.pfizer.com [148.168.224.84]) 17, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4PDqpcS027534 17, 31 -- for {microscopy-at-microscopy.com} ; Fri, 25 May 2007 08:52:51 -0500 17, 31 -- Received: from mopamrexc01.amer.pfizer.com (mopamrexc01.pfizer.com [170.116.32.254]) 17, 31 -- by gromsgoa01.pfizer.com (8.13.7/8.13.7) with ESMTP id l4PDqp7P016841; 17, 31 -- Fri, 25 May 2007 09:52:51 -0400 17, 31 -- Received: from groamrexc02.amer.pfizer.com ([172.30.8.169]) by mopamrexc01.amer.pfizer.com with Microsoft SMTPSVC(6.0.3790.1830); 17, 31 -- Fri, 25 May 2007 09:52:50 -0400 17, 31 -- Received: from anaamrexm01.amer.pfizer.com ([162.48.101.10]) by groamrexc02.amer.pfizer.com with Microsoft SMTPSVC(6.0.3790.1830); 17, 31 -- Fri, 25 May 2007 09:52:50 -0400 17, 31 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 17, 31 -- Content-class: urn:content-classes:message 17, 31 -- MIME-Version: 1.0 17, 31 -- Content-Type: text/plain; 17, 31 -- charset="us-ascii" 17, 31 -- Subject: RE: [Microscopy] Re: monkey perfusion methodology 17, 31 -- Date: Fri, 25 May 2007 09:52:49 -0400 17, 31 -- Message-ID: {CF89AF6ABD93B746A55BCCD9480A4B3D09284E76-at-anaamrexm01.amer.pfizer.com} 17, 31 -- In-Reply-To: {200705231748.l4NHmcPT011123-at-ns.microscopy.com} 17, 31 -- X-MS-Has-Attach: 17, 31 -- X-MS-TNEF-Correlator: 17, 31 -- Thread-Topic: [Microscopy] Re: monkey perfusion methodology 17, 31 -- Thread-Index: AcedYpv9QDW0Et4dTsemDk5MRB6YOwBcBQmA 17, 31 -- From: "Bobrowski, Walter" {Walter.Bobrowski-at-pfizer.com} 17, 31 -- To: {larry.ackerman-at-ucsf.edu} , {microscopy-at-microscopy.com} 17, 31 -- X-OriginalArrivalTime: 25 May 2007 13:52:50.0376 (UTC) FILETIME=[FBD64480:01C79ED3] 17, 31 -- X-Proofpoint-Virus-Version: vendor=fsecure engine=4.65.5502:2.3.11,1.2.37,4.0.164 definitions=2007-05-25_03:2007-05-25,2007-05-25,2007-05-25 signatures=0 17, 31 -- X-Proofpoint-Spam-Reason: safe 17, 31 -- Content-Transfer-Encoding: 8bit 17, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4PDqpcS027534 ==============================End of - Headers==============================
In fear if you have a conventional sputter coater - not the high vacuum versions that are designed for more difficult materials like chromium - you will not be able to coat other than with the simple materials - gold, platinum or gold-palladium.
Basically the everyday SEM sputter coater does not have the power.
Steve Chapman Senior Consultant Protrain For electron microscopy consultancy and training world wide Tel +44 1280 816512 Fax +44 1280 814007 Mobile +44 7711 606967 Web www.emcourses.com
----- Original Message ----- X-from: {david.wilbur-at-tufts.edu} To: {protrain-at-emcourses.com} Sent: Friday, May 25, 2007 2:05 PM
The convention of identifying magnification is one that seems to warrant some new attention in our digitized world. As several people have pointed out, there are several good conventions for reliably labeling the true scale for micrographs, the most common being the micron bar. That should probably remain the "gold standard" for publications, etc.
The familiar concept of "X" magnification is, of course, loaded with problems for digital images. It's not just the size of the instrument's viewing screen, but just what is the "magnification" of an image stored in a digital file? There has to be a point of reference, and historically that was the size of Polaroid film.
However, I think we should not casually dismiss the use of a "magnification" value as an identifier on our micrographs and our instruments. First of all, the use of "X" terminology is so solidly entrenched in our field that even if we don't like it, it's not going to go away anytime soon since it's just too convenient. The instrument's magnification knob has long been used as the means for setting up consistent operation, and was (in the Polaroid days) a pretty solid convention that you could count on -- a micrograph taken at 1000X on two different instruments would be generally comparable at least. Though I'm sure it's possible to write a procedure that specifies how to set the image scale in terms of the micron bar, it's much simpler to just say "set the control to 1000X." And if you're taking a series of micrographs at different magnifications, don't most of us label them like "10KX", "500X" etc.?
More fundamentally, it seems to me that the "X" concept speaks to something intrinsic about the way our brain perceives visual size. Think about a pair of binoculars that is specified as having 10X magnification. In that case, it is certainly not the absolute size of the recorded image that is being referenced (just what is the "size" of an image in one's mind?) but it relates to the perceived size of things. The convention for magnification in light optics is formally based on the angular deflection produced by the optics, but we perceive it as how much of the visual field is filled. I'll argue that the "X" value of SEM micrographs performs the same function. Thus, even if I see a "500X" micrograph reduced to a "thumbnail" or projected on a screen, I don't find it confusing since I'm gauging the size of the contents to the size of its frame. In other words, a term like "500X," though not sufficient for accurate size calibration in publications and the like, is still meaningful for making rapid assessments and comparisons, and I think we would all be well served if the convention of "X" magnification were better standardized. I'd propose two modest conventions:
(1) We adopt a standard terminology (perhaps "relative magnification") to refer to such "X" values, so everyone knows it is relative to the size of the field (not the viewing medium); and (2) We adopt a standard reference field size convention such as 100mm for the smaller field dimension (close to the four inch dimension of Polaroid film) to which an "X" value always refers.
This should not replace micron bars or other more quantitative metrics, but an agreed convention would, I think, eliminate a lot of confusion that currently exists.
Fred Schamber Aspex Corporation
-----Original Message----- X-from: bozzola-at-siu.edu [mailto:bozzola-at-siu.edu] Sent: Thursday, May 24, 2007 5:55 PM To: Fred Schamber
Magnification, as you have indicated, can be a dangerous concept especially when one deals with digital images (in TEM, SEM, and LM). In our beginning microscopy classes, we try to emphasize that the micrometer bar is what is to be displayed on the image, never the magnifaction. However, as a relative means of orientation, one could state in the legend that the original image was examined at a instrumental mangification of 10,000x, etc.
JB
} Hi Warren, } } I don't think that there was a "rule" that } changed. As long as the first document from a } microscope was relatively inflexible (a negative } or Polaroid), it was quite easy to go back and } get good numbers with information like "mag: } 10,000x". This already changed when people } started to use computers to look at images. If } you reduce the image size by a factor of 2 (to } insert in a publication, for example), you have } already gone from 10,000x to 5,000x. Now, with } digital acquisition and the flexibility to } acquire a certain number of "pixels", the } "magnification" has become a dangerous concept. } If you received an image in the mail and it says } "10,000x", you would have to know it's entire } history to judge the validity of the } magnification and make measurements. } } A better number is probably one that does not } assume a certain size of the final image to } specify dimensions. We used to specify the "full } width" of the image in microns or nm, today with } the digital images, it is better to specify the } dimension of a single pixel. That can then be } used to directly measure or to display a scale } bar, or even to calcualte the real magnification } on paper. } } The advantage of using magnification is that it } immediately gives you an idea of size. If you } look at something that was acquired at 100,000x, } you know that it must be small. } } } Michael Bode, Ph.D. } } General Manager } } OLYMPUS SOFT IMAGING SOLUTIONS } 12596 West Bayaud Ave #300 } Lakewood, CO 80228 } USA } Tel.: +1 (303) 234-9270 } Fax.: +1 (303) 234-9271 } E-mail: Mike.Bode-at-olympus-sis.com } www.olympus-sis.com } } -----Original Message----- } X-from: wesaia-at-iastate.edu [mailto:wesaia-at-iastate.edu] } Sent: Thursday, May 24, 2007 15:23 } To: Mike Bode } Subject: [Microscopy] Phenom-Ed SEM } } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The } Microscopy Society of America To } Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver
John J. Bozzola, Ph.D., Director Integrated Microscopy & Graphics Expertise (IMAGE) Southern Illinois University 750 Communications Drive - MC 4402 Carbondale, IL 62901 Telephone: 618-453-3730
Unless you have the scale bar right on an object, it will give you only a good idea of the size and dimensions of an object, but the same is true for "magnification". The difference is that a scale bar gives you an immediate idea of the real size of an object, while magnification is indirect and you have to convert it into a "virtual scale bar" first ("Let's see, the mag is 10,000x, so 1 cm on the image corresponds to 1 micron, so the object is roughly 4 microns long). When it comes to actual measurements, you have to take additional steps.
As far as edges for measurement go, Philip os of course right. SEMs in particular show edge effects that need to be modeled if accurate measurements are desired at high resolution.
mike
Michael Bode, Ph.D.
General Manager
OLYMPUS SOFT IMAGING SOLUTIONS 12596 West Bayaud Ave #300 Lakewood, CO 80228 USA Tel.: +1 (303) 234-9270 Fax.: +1 (303) 234-9271 E-mail: Mike.Bode-at-olympus-sis.com www.olympus-sis.com
-----Original Message----- X-from: oshel1pe-at-cmich.edu [mailto:oshel1pe-at-cmich.edu] Sent: Friday, May 25, 2007 06:58 To: Mike Bode
Stephane,
No, you don't know the size directly. Not unless the measured object is exactly orthogonal to the projected sight line. Quick experiment: project a bright light on the wall, and hold up a pencil (or similar). Hold the pencil so its long axis is exactly at 90 degrees to the central axis of the light path. Pretend the enlarged size of the pencil shadow is magnification. Now, tip the pencil into different orientations. Notice how the size of the shadow changes? Projective geometry. Same thing with an SEM image. It's a 3D object whose image is projected onto a 2D plane. Unless the dimension desired is exactly orthongonal to the line of sight, then the projected size will be different than the true size by an unknown amount. This can be solved by tilting the image, taking stereopairs, and doing trigonometry. But it will still be wrong unless the instrument has been properly calibrated *and* unless the all of the effects the specimen processing steps have on the sample are exactly known. Mind, this all ignores the more subtle question of "where's the edge"? Given the signal that forms an SEM image come from an interaction volume, not a dimensionless point, then the edge of a feature is essentially smeared out over the size of that volume. This is not much of an issue, or even no issue, for most imaging, but at high magnifications and resolutions, it is.
Phil
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==============================Original Headers============================== 4, 23 -- From oshel1pe-at-cmich.edu Fri May 25 07:53:43 2007 4, 23 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 4, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4PCrh4u022930 4, 23 -- for {Microscopy-at-microscopy.com} ; Fri, 25 May 2007 07:53:43 -0500 4, 23 -- Received: from egatea.central.cmich.local ([141.209.15.74]) 4, 23 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id l4PDGgqo010007; 4, 23 -- Fri, 25 May 2007 09:16:42 -0400 4, 23 -- Received: from [141.209.160.249] ([141.209.160.249]) by egatea.central.cmich.local with Microsoft SMTPSVC(6.0.3790.1830); 4, 23 -- Fri, 25 May 2007 08:53:41 -0400 4, 23 -- Mime-Version: 1.0 4, 23 -- Message-Id: {f06230904c27c89891143-at-[141.209.160.249]} 4, 23 -- In-Reply-To: {200705250804.l4P849BF030109-at-ns.microscopy.com} 4, 23 -- References: {200705250804.l4P849BF030109-at-ns.microscopy.com} 4, 23 -- Date: Fri, 25 May 2007 08:53:38 -0400 4, 23 -- To: nizets2-at-yahoo.com 4, 23 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 4, 23 -- Subject: Re: [Microscopy] Phenom-Ed SEM 4, 23 -- Cc: Microscopy-at-microscopy.com 4, 23 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 4, 23 -- X-OriginalArrivalTime: 25 May 2007 12:53:41.0660 (UTC) FILETIME=[B8A32DC0:01C79ECB] 4, 23 -- X-CanItPRO-Stream: default 4, 23 -- X-Spam-Score: -4 () L_EXCH_MF 4, 23 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
==============================Original Headers============================== 18, 25 -- From Mike.Bode-at-olympus-sis.com Fri May 25 10:24:22 2007 18, 25 -- Received: from mail.soft-imaging.de (ns.soft-imaging.de [62.180.61.130]) 18, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4PFOLoa031170 18, 25 -- for {Microscopy-at-microscopy.com} ; Fri, 25 May 2007 10:24:21 -0500 18, 25 -- Received: from muenster.olympus-sis.com (muenster.olympus-sis.com [10.26.20.9]) 18, 25 -- by mail.soft-imaging.de (8.13.3/8.13.3/SuSE Linux 0.7) with ESMTP id l4PFOOup016808 18, 25 -- for {Microscopy-at-microscopy.com} ; Fri, 25 May 2007 17:24:26 +0200 18, 25 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 18, 25 -- Content-class: urn:content-classes:message 18, 25 -- MIME-Version: 1.0 18, 25 -- Content-Type: text/plain; 18, 25 -- charset="us-ascii" 18, 25 -- Subject: RE: [Microscopy] Re: Phenom-Ed SEM 18, 25 -- Date: Fri, 25 May 2007 17:21:50 +0200 18, 25 -- Message-ID: {78B53BA1C5A2D9449EDA30A98800EC94965BCE-at-ms-s-gws.soft-imaging.net} 18, 25 -- In-Reply-To: {200705251257.l4PCvWLG031099-at-ns.microscopy.com} 18, 25 -- X-MS-Has-Attach: 18, 25 -- X-MS-TNEF-Correlator: 18, 25 -- Thread-Topic: [Microscopy] Re: Phenom-Ed SEM 18, 25 -- Thread-Index: AceezEQk9T9YybO7QHy85l+/cIUrjgAEsPjw 18, 25 -- References: {200705251257.l4PCvWLG031099-at-ns.microscopy.com} 18, 25 -- From: "Mike Bode" {Mike.Bode-at-olympus-sis.com} 18, 25 -- To: {Microscopy-at-microscopy.com} 18, 25 -- Content-Transfer-Encoding: 8bit 18, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4PFOLoa031170 ==============================End of - Headers==============================
I would greatly appreciate advice on the following: I have flat Epon embedded section sandwiched between Kapton (polyimide) film and Permanox plastic slide. It seems that excess of epoxy run over the slide sides and I have not been successful in film removal which I never encountered previously with Aclar films. Can anyone suggest an approach to release the section? Solvent, epoxy removal, dip in liquid nitrogen?
Thank you in advance,
Albina
-- MIKHAYLOVA,ALBINA, PhD Materials Science and Engineering University of Florida PO Box 116400 Gainesville, Florida 32611 Phone: (352) 392-6533 Fax: (352) 392-3771 E-Mail: amich-at-ufl.edu
==============================Original Headers============================== 6, 22 -- From amich-at-ufl.edu Fri May 25 10:48:10 2007 6, 22 -- Received: from smtp.ufl.edu (smtp01.osg.ufl.edu [128.227.74.149]) 6, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4PFmAx8010712 6, 22 -- for {Microscopy-at-microscopy.com} ; Fri, 25 May 2007 10:48:10 -0500 6, 22 -- Received: from osgjas04.cns.ufl.edu (osgjas04.cns.ufl.edu [128.227.74.134]) 6, 22 -- by smtp.ufl.edu (8.13.7/8.13.7/2.5.9) with ESMTP id l4PFm7Uj3956952 6, 22 -- for {Microscopy-at-microscopy.com} ; Fri, 25 May 2007 11:48:08 -0400 6, 22 -- Message-ID: {955813631.253271180108087892.JavaMail.osg-at-osgjas04.cns.ufl.edu} 6, 22 -- Date: Fri, 25 May 2007 11:48:07 -0400 (EDT) 6, 22 -- From: "MIKHAYLOVA,ALBINA" {amich-at-ufl.edu} 6, 22 -- To: Microscopy-at-microscopy.com 6, 22 -- Subject: flat section release 6, 22 -- MIME-Version: 1.0 6, 22 -- Content-Type: text/plain; format=flowed; charset=us-ascii 6, 22 -- Content-Transfer-Encoding: 7bit 6, 22 -- X-Mailer: GatorMail WebMail (http://GatorMail.sf.net/) 6, 22 -- X-Originating-IP: 74.230.140.89 [74.230.140.89] 6, 22 -- X-Greylist: Sender IP whitelisted, not delayed by milter-greylist-3.0rc3 (smtp.ufl.edu [128.227.74.56]); Fri, 25 May 2007 11:48:08 -0400 (EDT) 6, 22 -- X-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 6, 22 -- X-UFL-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 6, 22 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) 6, 22 -- X-UFL-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) ==============================End of - Headers==============================
Then resubmit your message to the listserver, and include a link to the URL for the image.
Make sure to include a micron marker or some other length calibration.
Do not send the image as an attachment.
In this manner, everyone will be able to make measurements for you.
regards,
Jim
PS: OoO away........
} From mail-at-ns.microscopy.com Thu May 24 19:08:35 2007 } Date: Thu, 24 May 2007 18:10:04 -0500 } To: jquinn-at-www.matscieng.sunysb.edu } From: vlk7-at-alfred.edu } Reply-to: vlk7-at-alfred.edu } X-Resent-From: "Microscopy Listserver" {microscopy-at-microscopy.com} } Subject: [Microscopy] AskAMicroscopist: help with Image-Pro plus } Errors-To: MicroscopyListSpamFilter-at-microscopy.com } X-lewp: MicroscopyListSpam NAGS } } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (vlk7-at-alfred.edu) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, May 24, 2007 at 09:18:07 } --------------------------------------------------------------------------- } } Email: vlk7-at-alfred.edu } Name: Victoria Knox } } Organization: Alfred University } } Education: Undergraduate College } } Location: Alfred, NY USA } } Question: I've been working with a program called Image-Pro plus to measure the diameter of micron-sized fibers from SEM micrographs. I need some assistance in how the program is measureing these fibers because I am getting inaccurate data that is off by at least 25% each time. I have tried to get the data using different measurements but it is still off. Any help you can provide would be greatly appreciated. } } --------------------------------------------------------------------------- } } ==============================Original Headers============================== } 7, 11 -- From zaluzec-at-ultra5.microscopy.com Thu May 24 18:09:34 2007 } 7, 11 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) } 7, 11 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4ON9WZT021861 } 7, 11 -- for {microscopy-at-microscopy.com} ; Thu, 24 May 2007 18:09:33 -0500 } 7, 11 -- Mime-Version: 1.0 } 7, 11 -- Message-Id: {p06240804c27bcb93a38f-at-[206.69.208.22]} } 7, 11 -- Date: Thu, 24 May 2007 18:09:32 -0500 } 7, 11 -- To: microscopy-at-microscopy.com } 7, 11 -- From: vlk7-at-alfred.edu (by way of Ask-A-Microscopist) } 7, 11 -- Subject: AskAMicroscopist: help with Image-Pro plus } 7, 11 -- Content-Type: text/plain; charset="us-ascii" } ==============================End of - Headers============================== }
==============================Original Headers============================== 13, 12 -- From jquinn-at-www.matscieng.sunysb.edu Fri May 25 11:50:12 2007 13, 12 -- Received: from www.matscieng.sunysb.edu (www.matscieng.sunysb.edu [129.49.36.33]) 13, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4PGoC0g023086 13, 12 -- for {microscopy-at-microscopy.com} ; Fri, 25 May 2007 11:50:12 -0500 13, 12 -- Received: (from jquinn-at-localhost) 13, 12 -- by www.matscieng.sunysb.edu (8.11.6/8.11.6) id l4PGmYv25112; 13, 12 -- Fri, 25 May 2007 12:48:34 -0400 13, 12 -- Date: Fri, 25 May 2007 12:48:34 -0400 13, 12 -- From: Jim Quinn {jquinn-at-www.matscieng.sunysb.edu} 13, 12 -- Message-Id: {200705251648.l4PGmYv25112-at-www.matscieng.sunysb.edu} 13, 12 -- To: microscopy-at-microscopy.com, vlk7-at-alfred.edu 13, 12 -- Subject: re: Fiber diameter ==============================End of - Headers==============================
Let me add a little to this discussion. Even if the coater would have a high vacuum, there still are issues with these three materials. All of them have an oxide coating on the targets. Ti and Al are very thin. Cr takes longer to build a thicker oxide coating. You have to sputter the oxide away before you can use it to deposit the film. Ti is a getter material and so is Al to a lesser degree. What this means is that the thin film deposited will act as a pump to pump reactive species such as O2, H2O, H2, from the vacuum system. So you have to sputter for a significant time prior to actually laying the film down so that these species are pumped from the chamber and your film on your substrate does not act as a getter pump. Ti and Al will instantaneously oxidize when the sample is brought up to air, but it will be a very thin protective coat. This is important with respect to the properties and the thickness of your film. Cr will take longer to fully oxidize, but it will unless protected by something like our SampleSaver(TM) container.
It would really help everything if your base pressure for your coater is at least in the 10^-7 torr range or better.
You might consider putting a top layer of a very thin Ti onto these other materials to act as a protective oxide layer.
Disclaimer: SBT makes and sells the IBS/e ion beam sputter and etch system and the SampleSaver(TM) storage container.
-Scott
Scott D. Walck, Ph.D. Technical Director South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
US Toll Free: 1-800-728-2233 Tel: (949) 492-2600 Fax: (949) 492-1499
www.southbaytech.com walck-at-southbaytech.com
-----Original Message----- X-from: dljones-at-bestweb.net [mailto:dljones-at-bestweb.net] Sent: Friday, May 25, 2007 6:36 AM To: Walck-at-SouthBayTech.com
Dave,
I am not familiar with the specific sputter coater you are using. So here are some questions. Do you know if it is a straight DC sputter coater or magnitron sputter coater? What is the pumping system, just a mechanical pump?
Most sputter coaters for SEM sample prep only use a mechanical pump for vacuum generation. This means you are only going to about 10-3 torr of vacuum before backfilling and then coating. For most thin film depositions, this is quite poor vacuum. Most thin film sputter coaters will go to somewhere around 10-6 to 10-8 torr before backfilling for sputter deposition. They also usually have a way to clean the substrate prior to deposition, which your little sputter coater I imagine does not have. The amount of contamination you will have, both on the first coat and then when switching between targets will be much larger than usually considered acceptable for thin film deposition.
All that being said, I don't see any reason you can't use your machine to deposit the materials you are mentioning, but I do question how useful the resulting layered structure will end up being. I would have to know a lot more about what the desired end product is to give you my thoughts on that.
dj
On Fri, 25 May 2007, david.wilbur-at-tufts.edu wrote:
} } } } ---------------------------------------------------------------------- } ------ The Microscopy ListServer -- CoSponsor: The Microscopy Society } of America To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } I have a Polaron SC-502 sputter coater in the SEM lab. One of the } Engineering faculty wants to coat samples with chromium, titanium and } aluminum for an application other than SEM. Assuming I can get } targets that fit the sputter coater, are there other issues that would } prevent using these other metals? } } Thank You } } Dave Wilbur } -- } __________________________________ } David J. Wilbur, Ph.D. } Instrumentation Specialist } Department of Chemistry } Tufts University } 62 Talbot Ave. } Medford, MA 02155 } voice: 617-627-2163 } Fax: 617-627-3443 } email: david.wilbur-at-tufts.edu } __________________________________ } } ==============================Original } Headers============================== } 3, 20 -- From david.wilbur-at-tufts.edu Fri May 25 08:04:11 2007 3, 20 -- } Received: from beech.usg.tufts.edu (beech.usg.tufts.edu [130.64.1.92]) } 3, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4PD4BCj002195 } 3, 20 -- for {Microscopy-at-Microscopy.Com} ; Fri, 25 May 2007 08:04:11 -0500 } 3, 20 -- Received: from dhcp-130-64-102-103.medford.tufts.edu ([130.64.102.103]:1120) } 3, 20 -- by beech.usg.tufts.edu with esmtps (TLSv1:AES256-SHA:256) } 3, 20 -- (Exim 4.60) } 3, 20 -- (envelope-from {david.wilbur-at-tufts.edu} ) } 3, 20 -- id 1HrZSl-0005gD-dz } 3, 20 -- for Microscopy-at-Microscopy.Com; Fri, 25 May 2007 09:04:11 -0400 } 3, 20 -- Message-ID: {4656DECA.3060604-at-tufts.edu} 3, 20 -- Date: Fri, } 25 May 2007 09:04:10 -0400 3, 20 -- From: David Wilbur } {david.wilbur-at-tufts.edu} 3, 20 -- User-Agent: Thunderbird 2.0.0.0 } (Windows/20070326) 3, 20 -- MIME-Version: 1.0 3, 20 -- To: Microscopy } Listserver {Microscopy-at-Microscopy.Com} 3, 20 -- Subject: [Microscopy] } Can I use my SEM sputter coater for metals other than 3, 20 -- gold? } 3, 20 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed } 3, 20 -- Content-Transfer-Encoding: 7bit } ==============================End of - } Headers============================== }
==============================Original Headers============================== 8, 19 -- From dljones-at-bestweb.net Fri May 25 08:32:22 2007 8, 19 -- Received: from vms044pub.verizon.net (vms044pub.verizon.net [206.46.252.44]) 8, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4PDWM7C004042 8, 19 -- for {Microscopy-at-microscopy.com} ; Fri, 25 May 2007 08:32:22 -0500 8, 19 -- Received: from localhost ([71.247.102.125]) 8, 19 -- by vms044.mailsrvcs.net (Sun Java System Messaging Server 6.2-6.01 (built Apr 8, 19 -- 3 2006)) with ESMTPA id {0JIL00536MXI59AA-at-vms044.mailsrvcs.net} for 8, 19 -- Microscopy-at-microscopy.com; Fri, 25 May 2007 08:32:11 -0500 (CDT) 8, 19 -- Date: Fri, 25 May 2007 09:33:09 -0400 (Eastern Daylight Time) 8, 19 -- From: "David L. Jones" {dljones-at-bestweb.net} 8, 19 -- Subject: Re: [Microscopy] Can I use my SEM sputter coater for metals other than 8, 19 -- In-reply-to: {200705251307.l4PD7gEs008393-at-ns.microscopy.com} 8, 19 -- X-X-Sender: dljones-at-pop-croton.bestweb.net 8, 19 -- To: david.wilbur-at-tufts.edu 8, 19 -- Cc: Microscopy-at-microscopy.com 8, 19 -- Message-id: {Pine.WNT.4.64.0705250918100.3128-at-H-F1} 8, 19 -- MIME-version: 1.0 8, 19 -- Content-type: TEXT/PLAIN; charset=US-ASCII; format=flowed 8, 19 -- References: {200705251307.l4PD7gEs008393-at-ns.microscopy.com} ==============================End of - Headers==============================
==============================Original Headers============================== 20, 21 -- From walck-at-southbaytech.com Fri May 25 12:24:52 2007 20, 21 -- Received: from flpi101.sbcis.sbc.com (flpi101.sbcis.sbc.com [207.115.20.70]) 20, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4PHOqUF003279 20, 21 -- for {Microscopy-at-microscopy.com} ; Fri, 25 May 2007 12:24:52 -0500 20, 21 -- X-ORBL: [64.169.217.123] 20, 21 -- Received: from dynamicbl8uno3 (adsl-64-169-217-123.dsl.lsan03.pacbell.net [64.169.217.123]) 20, 21 -- by flpi101.sbcis.sbc.com (8.13.8 out.dk.spool/8.13.8) with ESMTP id l4PHO4tw021420 20, 21 -- for {Microscopy-at-microscopy.com} ; Fri, 25 May 2007 10:24:05 -0700 20, 21 -- From: "Scott Walck" {walck-at-southbaytech.com} 20, 21 -- To: {Microscopy-at-microscopy.com} 20, 21 -- Subject: RE: [Microscopy] Re: Can I use my SEM sputter coater for metals other than 20, 21 -- Date: Fri, 25 May 2007 10:25:10 -0700 20, 21 -- Message-ID: {000501c79ef1$a5f62f30$7801a8c0-at-dynamicbl8uno3} 20, 21 -- MIME-Version: 1.0 20, 21 -- Content-Type: text/plain; 20, 21 -- charset="us-ascii" 20, 21 -- Content-Transfer-Encoding: 7bit 20, 21 -- X-Mailer: Microsoft Office Outlook 11 20, 21 -- In-Reply-To: {200705251335.l4PDZbwd010132-at-ns.microscopy.com} 20, 21 -- Thread-Index: Acee0ZR+OlWn4mrcSQiGzNBa+gDb9wAHlAWw 20, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 ==============================End of - Headers==============================
The Polaron SC-502 is a great old coater, but unfortunately it only has the ability to sputter Noble (non-oxidizing) Metals like Au, Pt and Pd. If you wish to sputter oxidizing metals like Al, Cr, Ir, etc., you require a turbo pumped coater such as the Emitech K575X, http://www.ebsstore.com/ecommerce/control/product/~category_id=F1/~product_id=K575X, which is manufactured by Quorum Technologies, the same Company that manufactures the Polaron Range of instruments.
Best regards, Mike Nesta
Michael R. Nesta Managing Director Energy Beam Sciences, Inc. Tel: 860 653-0411 Fax: 860 653-0422 MNesta-at-ebsciences.com www.ebsciences.com “ADDING BRILLIANCE TO YOUR VISION”
DISCLAIMER: Energy Beam Sciences, Inc. is the US distributor for Quorum Technologies products, which include the Polaron and Emitech ranges of SEM specimen preparation instruments.
david.wilbur-at-tufts.edu wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } I have a Polaron SC-502 sputter coater in the SEM lab. One of the } Engineering faculty wants to coat samples with chromium, titanium and } aluminum for an application other than SEM. Assuming I can get targets } that fit the sputter coater, are there other issues that would prevent } using these other metals? } } Thank You } } Dave Wilbur }
==============================Original Headers============================== 10, 27 -- From mnesta-at-ebsciences.com Fri May 25 12:43:08 2007 10, 27 -- Received: from mx-outbound01.easydns.com (mailout.easydns.com [205.210.42.54]) 10, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4PHh8LF014952 10, 27 -- for {microscopy-at-microscopy.com} ; Fri, 25 May 2007 12:43:08 -0500 10, 27 -- Received: from mail.ebsciences.com (unknown [69.182.224.2]) 10, 27 -- (using TLSv1 with cipher DHE-RSA-AES256-SHA (256/256 bits)) 10, 27 -- (No client certificate requested) 10, 27 -- by mx-outbound01.easydns.com (Postfix) with ESMTP id EE8CB80B3; 10, 27 -- Fri, 25 May 2007 13:43:06 -0400 (EDT) 10, 27 -- Received: from mnesta.ebsciences.private ([10.10.0.153]) 10, 27 -- by mail.ebsciences.com with esmtpsa (TLSv1:AES256-SHA:256) 10, 27 -- (Exim 4.67) 10, 27 -- (envelope-from {mnesta-at-ebsciences.com} ) 10, 27 -- id 1Hrdoh-00036G-34; Fri, 25 May 2007 13:43:07 -0400 10, 27 -- Message-ID: {46572029.1030903-at-ebsciences.com} 10, 27 -- Date: Fri, 25 May 2007 13:43:05 -0400 10, 27 -- From: Mike Nesta {mnesta-at-ebsciences.com} 10, 27 -- Organization: Energy Beam Sciences 10, 27 -- User-Agent: Thunderbird 1.5.0.10 (Windows/20070221) 10, 27 -- MIME-Version: 1.0 10, 27 -- To: david.wilbur-at-tufts.edu, microscopy-at-microscopy.com 10, 27 -- Subject: Re: [Microscopy] Can I use my SEM sputter coater for metals other 10, 27 -- than 10, 27 -- References: {200705251307.l4PD7CNq007145-at-ns.microscopy.com} 10, 27 -- In-Reply-To: {200705251307.l4PD7CNq007145-at-ns.microscopy.com} 10, 27 -- Content-Type: text/plain; charset=windows-1252; format=flowed 10, 27 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
Aluminum and titanium will deposit as alumina and titania unless you've got a turbopump or better. There is no easy solution. Even then, it will instantly oxidize upon exposure to air.
If the SC-502 is a magnetron sputter unit, then the chrome may shunt the field.
regards,
Jim
} From mail-at-ns.microscopy.com Fri May 25 09:03:20 2007 } Date: Fri, 25 May 2007 08:04:51 -0500 } To: jquinn-at-www.matscieng.sunysb.edu } From: david.wilbur-at-tufts.edu } Reply-to: david.wilbur-at-tufts.edu } X-Resent-From: "Microscopy Listserver" {microscopy-at-microscopy.com} } Subject: [Microscopy] Can I use my SEM sputter coater for metals other than } Errors-To: MicroscopyListSpamFilter-at-microscopy.com } X-lewp: MicroscopyListSpam NAGS } } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } I have a Polaron SC-502 sputter coater in the SEM lab. One of the } Engineering faculty wants to coat samples with chromium, titanium and } aluminum for an application other than SEM. Assuming I can get targets } that fit the sputter coater, are there other issues that would prevent } using these other metals? } } Thank You } } Dave Wilbur } -- } __________________________________ } David J. Wilbur, Ph.D. } Instrumentation Specialist } Department of Chemistry } Tufts University } 62 Talbot Ave. } Medford, MA 02155 } voice: 617-627-2163 } Fax: 617-627-3443 } email: david.wilbur-at-tufts.edu } __________________________________ } } ==============================Original Headers============================== } 3, 20 -- From david.wilbur-at-tufts.edu Fri May 25 08:04:11 2007 } 3, 20 -- Received: from beech.usg.tufts.edu (beech.usg.tufts.edu [130.64.1.92]) } 3, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4PD4BCj002195 } 3, 20 -- for {Microscopy-at-Microscopy.Com} ; Fri, 25 May 2007 08:04:11 -0500 } 3, 20 -- Received: from dhcp-130-64-102-103.medford.tufts.edu ([130.64.102.103]:1120) } 3, 20 -- by beech.usg.tufts.edu with esmtps (TLSv1:AES256-SHA:256) } 3, 20 -- (Exim 4.60) } 3, 20 -- (envelope-from {david.wilbur-at-tufts.edu} ) } 3, 20 -- id 1HrZSl-0005gD-dz } 3, 20 -- for Microscopy-at-Microscopy.Com; Fri, 25 May 2007 09:04:11 -0400 } 3, 20 -- Message-ID: {4656DECA.3060604-at-tufts.edu} } 3, 20 -- Date: Fri, 25 May 2007 09:04:10 -0400 } 3, 20 -- From: David Wilbur {david.wilbur-at-tufts.edu} } 3, 20 -- User-Agent: Thunderbird 2.0.0.0 (Windows/20070326) } 3, 20 -- MIME-Version: 1.0 } 3, 20 -- To: Microscopy Listserver {Microscopy-at-Microscopy.Com} } 3, 20 -- Subject: [Microscopy] Can I use my SEM sputter coater for metals other than } 3, 20 -- gold? } 3, 20 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed } 3, 20 -- Content-Transfer-Encoding: 7bit } ==============================End of - Headers============================== }
==============================Original Headers============================== 7, 12 -- From jquinn-at-www.matscieng.sunysb.edu Fri May 25 12:43:10 2007 7, 12 -- Received: from www.matscieng.sunysb.edu (www.matscieng.sunysb.edu [129.49.36.33]) 7, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4PHh91S014974 7, 12 -- for {microscopy-at-microscopy.com} ; Fri, 25 May 2007 12:43:10 -0500 7, 12 -- Received: (from jquinn-at-localhost) 7, 12 -- by www.matscieng.sunysb.edu (8.11.6/8.11.6) id l4PHfVU25277 7, 12 -- for microscopy-at-microscopy.com; Fri, 25 May 2007 13:41:31 -0400 7, 12 -- Date: Fri, 25 May 2007 13:41:31 -0400 7, 12 -- From: Jim Quinn {jquinn-at-www.matscieng.sunysb.edu} 7, 12 -- Message-Id: {200705251741.l4PHfVU25277-at-www.matscieng.sunysb.edu} 7, 12 -- To: microscopy-at-microscopy.com 7, 12 -- Subject: re: Can I use my SEM sputter coater for metals other than ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (vlk7-at-alfred.edu) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, May 25, 2007 at 12:19:24 ---------------------------------------------------------------------------
Email: vlk7-at-alfred.edu Name: Victoria Knox
Organization: Alfred University
Education: Undergraduate College
Location: Alfred, NY USA
Question: This is a re-post with a URL to an example image in question:
I've been working with a program called Image-Pro plus to measure the diameter of micron-sized fibers from SEM micrographs. I need some assistance in how the program is measureing these fibers because I am getting inaccurate data that is off by at least 25% each time. I have tried to get the data using different measurements but it is still off. Any help you can provide would be greatly appreciated.
Your magnification is too low. The answer is that simple.
If the fiber size is only a few pixels wide, then how will you get good results?
Also, I assume that you have use a spatial calibration.
Jim
} From mail-at-ns.microscopy.com Fri May 25 14:36:32 2007 } Date: Fri, 25 May 2007 13:38:05 -0500 } To: jquinn-at-www.matscieng.sunysb.edu } From: vlk7-at-alfred.edu } Reply-to: vlk7-at-alfred.edu } X-Resent-From: "Microscopy Listserver" {microscopy-at-microscopy.com} } Subject: [Microscopy] AskAMicroscopist: Image Pro Analysis Question - URL with an Image } Errors-To: MicroscopyListSpamFilter-at-microscopy.com } X-lewp: MicroscopyListSpam NAGS } } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (vlk7-at-alfred.edu) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, May 25, 2007 at 12:19:24 } --------------------------------------------------------------------------- } } Email: vlk7-at-alfred.edu } Name: Victoria Knox } } Organization: Alfred University } } Education: Undergraduate College } } Location: Alfred, NY USA } } Question: This is a re-post with a URL to an example image in question: } } http://i100.photobucket.com/albums/m16/ceramicchick7/362M-B.jpg } } I've been working with a program called Image-Pro plus to measure the diameter of micron-sized fibers from SEM micrographs. I need some assistance in how the program is measureing these fibers because I am getting inaccurate data that is off by at least 25% each time. I have tried to get the data using different measurements but it is still off. Any help you can provide would be greatly appreciated. } } --------------------------------------------------------------------------- } } ==============================Original Headers============================== } 9, 11 -- From zaluzec-at-ultra5.microscopy.com Fri May 25 13:37:16 2007 } 9, 11 -- Received: from [206.69.208.22] (msdvpn8.msd.anl.gov [130.202.238.72]) } 9, 11 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4PIbDcZ006448 } 9, 11 -- for {microscopy-at-microscopy.com} ; Fri, 25 May 2007 13:37:15 -0500 } 9, 11 -- Mime-Version: 1.0 } 9, 11 -- Message-Id: {p06240812c27cdd0dbd57-at-[206.69.208.22]} } 9, 11 -- Date: Fri, 25 May 2007 13:37:13 -0500 } 9, 11 -- To: microscopy-at-microscopy.com } 9, 11 -- From: vlk7-at-alfred.edu (by way of Ask-A-Microscopist) } 9, 11 -- Subject: AskAMicroscopist: Image Pro Analysis Question - URL with an Image } 9, 11 -- Content-Type: text/plain; charset="us-ascii" } ==============================End of - Headers============================== }
==============================Original Headers============================== 10, 12 -- From jquinn-at-www.matscieng.sunysb.edu Fri May 25 14:00:16 2007 10, 12 -- Received: from www.matscieng.sunysb.edu (www.matscieng.sunysb.edu [129.49.36.33]) 10, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4PJ0G43018466 10, 12 -- for {microscopy-at-microscopy.com} ; Fri, 25 May 2007 14:00:16 -0500 10, 12 -- Received: (from jquinn-at-localhost) 10, 12 -- by www.matscieng.sunysb.edu (8.11.6/8.11.6) id l4PIwbT25434; 10, 12 -- Fri, 25 May 2007 14:58:37 -0400 10, 12 -- Date: Fri, 25 May 2007 14:58:37 -0400 10, 12 -- From: Jim Quinn {jquinn-at-www.matscieng.sunysb.edu} 10, 12 -- Message-Id: {200705251858.l4PIwbT25434-at-www.matscieng.sunysb.edu} 10, 12 -- To: microscopy-at-microscopy.com, vlk7-at-alfred.edu 10, 12 -- Subject: Re: [Microscopy] AskAMicroscopist: Image Pro Analysis Question - URL with an Image ==============================End of - Headers==============================
A good rule of thumb in image analysis is to have about 30 features in each field of view that you are counting. However, some of that depends on the degree of dispersion (separation) of the features, their sizes, and in your case the aspect ratios. If you have a very good dispersion of smaller (rounder) features, then higher numbers of features can be counted per field but you risk flocculation affects.
Raise the mag to start with and count multiple fields of view.
Paul
At 02:00 PM 5/25/07 -0500, you wrote: } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 5, 26 -- From beaurega-at-westol.com Fri May 25 17:22:44 2007 5, 26 -- Received: from smtp-gateway-2.winbeam.com (smtp-gateway-2.winbeam.com [64.84.97.67]) 5, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4PMMiDB012478 5, 26 -- for {microscopy-at-microscopy.com} ; Fri, 25 May 2007 17:22:44 -0500 5, 26 -- Received: from mail.winbeam.com (mail.winbeam.com [64.84.96.10]) 5, 26 -- by smtp-gateway-2.winbeam.com (8.13.1/8.12.8) with SMTP id l4PMJJsr014493 5, 26 -- for {microscopy-at-microscopy.com} ; Fri, 25 May 2007 18:19:20 -0400 5, 26 -- Received: (qmail 10736 invoked by uid 89); 25 May 2007 22:19:17 -0000 5, 26 -- Received: from pitts-69-72-109-13.dynamic-dialup.coretel.net (HELO beaurega) (69.72.109.13) 5, 26 -- by mail.winbeam.com with SMTP; 25 May 2007 22:19:17 -0000 5, 26 -- Message-Id: {3.0.6.32.20070525181922.007a08b0-at-pop3.norton.antivirus} 5, 26 -- X-Sender: beaurega/mail.westol.com-at-pop3.norton.antivirus 5, 26 -- X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.6 (32) 5, 26 -- Date: Fri, 25 May 2007 18:19:22 -0400 5, 26 -- To: jquinn-at-www.matscieng.sunysb.edu, microscopy-at-microscopy.com 5, 26 -- From: Beaurega {beaurega-at-westol.com} 5, 26 -- Subject: Re: [Microscopy] Re: AskAMicroscopist: Image Pro Analysis 5, 26 -- Question - URL with an Image 5, 26 -- In-Reply-To: {200705251900.l4PJ0den019188-at-ns.microscopy.com} 5, 26 -- Mime-Version: 1.0 5, 26 -- Content-Type: text/plain; charset="us-ascii" 5, 26 -- X--MailScanner-Information: - Please contact Technical Support for more information 5, 26 -- X--MailScanner: Found to be clean (courtesy of MailScanner) 5, 26 -- X--MailScanner-SpamCheck: not spam (whitelisted), SpamAssassin (not cached, 5, 26 -- timed out) 5, 26 -- X--MailScanner-From: beaurega-at-westol.com ==============================End of - Headers==============================
However, seeing your image, you could try an automated approach for your measurements. You could have the software detect the fibers and measure their areas, then do a skeletonization and calculate the length. Divide the area by the length, and you'll have a measurement of the width. If you do that for multiple fibers, you should get some reasonable statistics of the fiber width and length. Unfortunately I do not know how that works in Image-Pro.
mike
Michael Bode, Ph.D.
General Manager
OLYMPUS SOFT IMAGING SOLUTIONS 12596 West Bayaud Ave #300 Lakewood, CO 80228 USA Tel.: +1 (303) 234-9270 Fax.: +1 (303) 234-9271 E-mail: Mike.Bode-at-olympus-sis.com www.olympus-sis.com
-----Original Message----- X-from: beaurega-at-westol.com [mailto:beaurega-at-westol.com] Sent: Friday, May 25, 2007 16:28 To: Mike Bode
I agree with Jim after seeing the image.
A good rule of thumb in image analysis is to have about 30 features in each field of view that you are counting. However, some of that depends on the degree of dispersion (separation) of the features, their sizes, and in your case the aspect ratios. If you have a very good dispersion of smaller (rounder) features, then higher numbers of features can be counted per field but you risk flocculation affects.
Raise the mag to start with and count multiple fields of view.
Paul
At 02:00 PM 5/25/07 -0500, you wrote: } } } } ----------------------------------------------------------------------- } ----- The Microscopy ListServer -- CoSponsor: The Microscopy Society } of America To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver
} results? } } Also, I assume that you have use a spatial calibration. } } Jim } } } } From mail-at-ns.microscopy.com Fri May 25 14:36:32 2007 } } Date: Fri, 25 May 2007 13:38:05 -0500 } } To: jquinn-at-www.matscieng.sunysb.edu } } From: vlk7-at-alfred.edu } } Reply-to: vlk7-at-alfred.edu } } X-Resent-From: "Microscopy Listserver" {microscopy-at-microscopy.com} } } Subject: [Microscopy] AskAMicroscopist: Image Pro Analysis Question } } - URL with an Image } } Errors-To: MicroscopyListSpamFilter-at-microscopy.com } } X-lewp: MicroscopyListSpam NAGS } } } } } } } } } } ------------------------------------------------------------------------ ---- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } } America To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ------------------------------------------------------------------------ ---- } } } } Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (vlk7-at-alfred.edu) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, May 25, 2007 at 12:19:24 } } ------------------------------------------------------------------------ --- } } } } Email: vlk7-at-alfred.edu } } Name: Victoria Knox } } } } Organization: Alfred University } } } } Education: Undergraduate College } } } } Location: Alfred, NY USA } } } } Question: This is a re-post with a URL to an example image in question: } } } } http://i100.photobucket.com/albums/m16/ceramicchick7/362M-B.jpg } } } } I've been working with a program called Image-Pro plus to measure } } the diameter of } micron-sized fibers from SEM micrographs. I need some assistance in how the program } is measureing these fibers because I am getting inaccurate data that is off by } at least 25% each time. I have tried to get the data using different measurements } but it is still off. Any help you can provide would be greatly appreciated. } } } } ------------------------------------------------------------------------ --- } } } } ==============================Original Headers============================== } } 9, 11 -- From zaluzec-at-ultra5.microscopy.com Fri May 25 13:37:16 2007
} } 9, 11 -- Received: from [206.69.208.22] (msdvpn8.msd.anl.gov [130.202.238.72]) } } 9, 11 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4PIbDcZ006448 } } 9, 11 -- for {microscopy-at-microscopy.com} ; Fri, 25 May 2007 13:37:15 -0500 } } 9, 11 -- Mime-Version: 1.0 } } 9, 11 -- Message-Id: {p06240812c27cdd0dbd57-at-[206.69.208.22]} } } 9, 11 -- Date: Fri, 25 May 2007 13:37:13 -0500 9, 11 -- To: } } microscopy-at-microscopy.com 9, 11 -- From: vlk7-at-alfred.edu (by way of } } Ask-A-Microscopist) 9, 11 -- Subject: AskAMicroscopist: Image Pro } } Analysis Question - URL with an Image } } 9, 11 -- Content-Type: text/plain; charset="us-ascii" } } ==============================End of - Headers============================== } } } } ==============================Original } Headers============================== } 10, 12 -- From jquinn-at-www.matscieng.sunysb.edu Fri May 25 14:00:16 2007
} 10, 12 -- Received: from www.matscieng.sunysb.edu (www.matscieng.sunysb.edu [129.49.36.33]) } 10, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4PJ0G43018466 } 10, 12 -- for {microscopy-at-microscopy.com} ; Fri, 25 May 2007 14:00:16 -0500 } 10, 12 -- Received: (from jquinn-at-localhost) } 10, 12 -- by www.matscieng.sunysb.edu (8.11.6/8.11.6) id l4PIwbT25434; } 10, 12 -- Fri, 25 May 2007 14:58:37 -0400 } 10, 12 -- Date: Fri, 25 May 2007 14:58:37 -0400 10, 12 -- From: Jim } Quinn {jquinn-at-www.matscieng.sunysb.edu} 10, 12 -- Message-Id: } {200705251858.l4PIwbT25434-at-www.matscieng.sunysb.edu} } 10, 12 -- To: microscopy-at-microscopy.com, vlk7-at-alfred.edu 10, 12 -- } Subject: Re: [Microscopy] AskAMicroscopist: Image Pro Analysis Question - URL with an Image } ==============================End of - } Headers============================== } } }
==============================Original Headers============================== 5, 26 -- From beaurega-at-westol.com Fri May 25 17:22:44 2007 5, 26 -- Received: from smtp-gateway-2.winbeam.com (smtp-gateway-2.winbeam.com [64.84.97.67]) 5, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4PMMiDB012478 5, 26 -- for {microscopy-at-microscopy.com} ; Fri, 25 May 2007 17:22:44 -0500 5, 26 -- Received: from mail.winbeam.com (mail.winbeam.com [64.84.96.10]) 5, 26 -- by smtp-gateway-2.winbeam.com (8.13.1/8.12.8) with SMTP id l4PMJJsr014493 5, 26 -- for {microscopy-at-microscopy.com} ; Fri, 25 May 2007 18:19:20 -0400 5, 26 -- Received: (qmail 10736 invoked by uid 89); 25 May 2007 22:19:17 -0000 5, 26 -- Received: from pitts-69-72-109-13.dynamic-dialup.coretel.net (HELO beaurega) (69.72.109.13) 5, 26 -- by mail.winbeam.com with SMTP; 25 May 2007 22:19:17 -0000 5, 26 -- Message-Id: {3.0.6.32.20070525181922.007a08b0-at-pop3.norton.antivirus} 5, 26 -- X-Sender: beaurega/mail.westol.com-at-pop3.norton.antivirus 5, 26 -- X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.6 (32) 5, 26 -- Date: Fri, 25 May 2007 18:19:22 -0400 5, 26 -- To: jquinn-at-www.matscieng.sunysb.edu, microscopy-at-microscopy.com 5, 26 -- X-from: Beaurega {beaurega-at-westol.com} 5, 26 -- Subject: Re: [Microscopy] Re: AskAMicroscopist: Image Pro Analysis 5, 26 -- Question - URL with an Image 5, 26 -- In-Reply-To: {200705251900.l4PJ0den019188-at-ns.microscopy.com} 5, 26 -- Mime-Version: 1.0 5, 26 -- Content-Type: text/plain; charset="us-ascii" 5, 26 -- X--MailScanner-Information: - Please contact Technical Support for more information 5, 26 -- X--MailScanner: Found to be clean (courtesy of MailScanner) 5, 26 -- X--MailScanner-SpamCheck: not spam (whitelisted), SpamAssassin (not cached, 5, 26 -- timed out) 5, 26 -- X--MailScanner-From: beaurega-at-westol.com ==============================End of - Headers==============================
==============================Original Headers============================== 20, 25 -- From Mike.Bode-at-olympus-sis.com Fri May 25 18:12:02 2007 20, 25 -- Received: from mail.soft-imaging.de (ns.soft-imaging.de [62.180.61.130]) 20, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4PNC1gI024587 20, 25 -- for {Microscopy-at-microscopy.com} ; Fri, 25 May 2007 18:12:01 -0500 20, 25 -- Received: from muenster.olympus-sis.com (muenster.olympus-sis.com [10.26.20.9]) 20, 25 -- by mail.soft-imaging.de (8.13.3/8.13.3/SuSE Linux 0.7) with ESMTP id l4PNC8LV025181 20, 25 -- for {Microscopy-at-microscopy.com} ; Sat, 26 May 2007 01:12:10 +0200 20, 25 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 20, 25 -- Content-class: urn:content-classes:message 20, 25 -- MIME-Version: 1.0 20, 25 -- Content-Type: text/plain; 20, 25 -- charset="us-ascii" 20, 25 -- Subject: RE: [Microscopy] AskAMicroscopist: Image Pro Analysis 20, 25 -- Date: Sat, 26 May 2007 01:09:08 +0200 20, 25 -- Message-ID: {78B53BA1C5A2D9449EDA30A98800EC94965C5F-at-ms-s-gws.soft-imaging.net} 20, 25 -- In-Reply-To: {200705252228.l4PMS4JZ020552-at-ns.microscopy.com} 20, 25 -- X-MS-Has-Attach: 20, 25 -- X-MS-TNEF-Correlator: 20, 25 -- Thread-Topic: [Microscopy] AskAMicroscopist: Image Pro Analysis 20, 25 -- Thread-Index: AcefG/gtZvcEEtAfTKaK1nOu8A3PAwAA4r7w 20, 25 -- References: {200705252228.l4PMS4JZ020552-at-ns.microscopy.com} 20, 25 -- From: "Mike Bode" {Mike.Bode-at-olympus-sis.com} 20, 25 -- To: {Microscopy-at-microscopy.com} 20, 25 -- Content-Transfer-Encoding: 8bit 20, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4PNC1gI024587 ==============================End of - Headers==============================
I am not sure to understand what you mean. The electron beam IS orthogonal to the object visualized. For the "edge smearing" I can understand the problem though.
Stephane
--- Philip Oshel {oshel1pe-at-cmich.edu} wrote:
} Stephane, } } No, you don't know the size directly. Not unless the } measured object } is exactly orthogonal to the projected sight line. } Quick experiment: project a bright light on the } wall, and hold up a } pencil (or similar). Hold the pencil so its long } axis is exactly at } 90 degrees to the central axis of the light path. } Pretend the } enlarged size of the pencil shadow is magnification. } Now, tip the pencil into different orientations. } Notice how the size } of the shadow changes? Projective geometry. } Same thing with an SEM image. It's a 3D object whose } image is } projected onto a 2D plane. Unless the dimension } desired is exactly } orthongonal to the line of sight, then the projected } size will be } different than the true size by an unknown amount. } This can be solved by tilting the image, taking } stereopairs, and } doing trigonometry. } But it will still be wrong unless the instrument has } been properly } calibrated *and* unless the all of the effects the } specimen } processing steps have on the sample are exactly } known. } Mind, this all ignores the more subtle question of } "where's the } edge"? Given the signal that forms an SEM image come } from an } interaction volume, not a dimensionless point, then } the edge of a } feature is essentially smeared out over the size of } that volume. This } is not much of an issue, or even no issue, for most } imaging, but at } high magnifications and resolutions, it is. } } Phil } } } ---------------------------------------------------------------------------- } } The Microscopy ListServer -- CoSponsor: The } Microscopy Society of America } } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------------- } } } } Sure, but if you see a scale bar next to your } } feature, you don't know it is small, you know its } size } } directly! (and if the full scale bar is 50 nm, you } } know it is } } very small) } } } } Stephane } } } } } } } --- Mike.Bode-at-olympus-sis.com wrote: } } } } } } } } } } } The advantage of using magnification is that } it } } } } immediately gives you an idea of size. If you } look } } } } at something that was acquired at 100,000x, } you } } } know } } } } that it must be small. } } } } } } } } } } } } Michael Bode, Ph.D. } } } } } } } } General Manager } } } } } } } } OLYMPUS SOFT IMAGING SOLUTIONS } } } } 12596 West Bayaud Ave #300 } } } } Lakewood, CO 80228 } } } } USA } } } } Tel.: +1 (303) 234-9270 } } } } Fax.: +1 (303) 234-9271 } } } } E-mail: Mike.Bode-at-olympus-sis.com } } } } www.olympus-sis.com } -- } Philip Oshel } Microscopy Facility Supervisor } Biology Department } 024C Brooks Hall } Central Michigan University } Mt. Pleasant, MI 48859 }
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==============================Original Headers============================== 9, 21 -- From nizets2-at-yahoo.com Sat May 26 01:47:11 2007 9, 21 -- Received: from web37410.mail.mud.yahoo.com (web37410.mail.mud.yahoo.com [209.191.91.142]) 9, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l4Q6lBJO010693 9, 21 -- for {microscopy-at-microscopy.com} ; Sat, 26 May 2007 01:47:11 -0500 9, 21 -- Received: (qmail 55975 invoked by uid 60001); 26 May 2007 06:47:10 -0000 9, 21 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 9, 21 -- s=s1024; d=yahoo.com; 9, 21 -- h=X-YMail-OSG:Received:Date:From:Subject:To:Cc:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding:Message-ID; 9, 21 -- b=njncCplQkToC1dbFVtybi/g8ulmAz5d5FWtAhsEI27GDyg/38YbGIQh4o48TFSCwJmagr8WWAIRxbO9z3JAXKCFPcuFij8D+ZthO2EPhKvX7hDiwN2pg8HEoS3E9g7zLJZ5z5f+uUMFA6KEeKKi+GowxJpxX02R1GZPl6HLypi0=; 9, 21 -- X-YMail-OSG: HPblcL8VM1nWZcLb1xJ6kKpWBIHVI.OvhItK1GbYdtMPwRDDiNAfV27GdyDNqEalmyJX49jkEhvzQQ0z7vz.eKwB8vOM.Bi3fReYsv15CcWkxIkmxLw- 9, 21 -- Received: from [80.121.36.211] by web37410.mail.mud.yahoo.com via HTTP; Fri, 25 May 2007 23:47:10 PDT 9, 21 -- Date: Fri, 25 May 2007 23:47:10 -0700 (PDT) 9, 21 -- From: Stephane Nizet {nizets2-at-yahoo.com} 9, 21 -- Subject: Re: [Microscopy] Phenom-Ed SEM 9, 21 -- To: Philip Oshel {oshel1pe-at-cmich.edu} 9, 21 -- Cc: microscopy-at-microscopy.com 9, 21 -- In-Reply-To: {f06230904c27c89891143-at-[141.209.160.249]} 9, 21 -- MIME-Version: 1.0 9, 21 -- Content-Type: text/plain; charset=iso-8859-1 9, 21 -- Content-Transfer-Encoding: 8bit 9, 21 -- Message-ID: {629527.55003.qm-at-web37410.mail.mud.yahoo.com} ==============================End of - Headers==============================
Remember when the subject has drifting off the original topic, as it has in the discussion on of the FEI table top SEM, into a new area it would be advisable to modify the subject line so that it reflects the discussion. It will help others (as well as the search engine) in the long run.
Nestor Your FriendlyNeighborhood SysOp
==============================Original Headers============================== 5, 13 -- From zaluzec-at-microscopy.com Sat May 26 09:28:33 2007 5, 13 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) 5, 13 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4QESWip031249 5, 13 -- for {microscopy-at-microscopy.com} ; Sat, 26 May 2007 09:28:33 -0500 5, 13 -- Mime-Version: 1.0 5, 13 -- Message-Id: {p06240818c27df355f78e-at-[206.69.208.22]} 5, 13 -- In-Reply-To: {200705260647.l4Q6lBgL010709-at-ns.microscopy.com} 5, 13 -- References: {200705260647.l4Q6lBgL010709-at-ns.microscopy.com} 5, 13 -- Date: Sat, 26 May 2007 09:28:32 -0500 5, 13 -- To: microscopy-at-microscopy.com 5, 13 -- From: "Nestor J. Zaluzec" {zaluzec-at-microscopy.com} 5, 13 -- Subject: Administrivia: Subject Line / Topic Drift 5, 13 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
Just out of curiosity, what should the vacuum level be to get a steady electron beam in an older SEM? I grabbed a vacuum gauge off of my sputter coater and measured the vacuum at 1.2 x 10e-5. I don't think that's enough, though. Any thoughts?
--Justin.
==============================Original Headers============================== 2, 26 -- From kraftpiano-at-gmail.com Sat May 26 14:28:06 2007 2, 26 -- Received: from wa-out-1112.google.com (wa-out-1112.google.com [209.85.146.182]) 2, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4QJS6sc018284 2, 26 -- for {microscopy-at-microscopy.com} ; Sat, 26 May 2007 14:28:06 -0500 2, 26 -- Received: by wa-out-1112.google.com with SMTP id j5so466378wah 2, 26 -- for {microscopy-at-microscopy.com} ; Sat, 26 May 2007 12:28:06 -0700 (PDT) 2, 26 -- DKIM-Signature: a=rsa-sha1; c=relaxed/relaxed; 2, 26 -- d=gmail.com; s=beta; 2, 26 -- h=domainkey-signature:received:received:message-id:date:from:to:subject:mime-version:content-type:content-transfer-encoding:content-disposition; 2, 26 -- b=mF1yFAt4zQgEOCJKeNzwygekp8ZE/db54wXobsSKwofa2CazIlAwgbw25uvo8xGCbiB3/nNQSFxHzx4K2HBERScwHErSxVYTAj0Kq4Iex4H8VA6Dz52XgdYnMyea82zokTmhgON7TZTSJjI+iFzXsmxuXgm4ULI+Oyai7gHu+Jk= 2, 26 -- DomainKey-Signature: a=rsa-sha1; c=nofws; 2, 26 -- d=gmail.com; s=beta; 2, 26 -- h=received:message-id:date:from:to:subject:mime-version:content-type:content-transfer-encoding:content-disposition; 2, 26 -- b=g2BLT42OeNHc/SIs4cL4WfBilfLyzgUf6maiRBhhc3w5bSovMX9arWLdkVH6inU4QgO6jPiSCiwWkE/R5Lq0c8VuBBVAWsPShwHhcdm49/jJOaBQfSHJesKY04Xn+8UjGp7X6mpmr+VkdSUMXW5vdFFC/gqvhue/32AkzV6bFKs= 2, 26 -- Received: by 10.114.14.1 with SMTP id 1mr2063238wan.1180207686080; 2, 26 -- Sat, 26 May 2007 12:28:06 -0700 (PDT) 2, 26 -- Received: by 10.114.78.15 with HTTP; Sat, 26 May 2007 12:28:06 -0700 (PDT) 2, 26 -- Message-ID: {25e2b0d20705261228g395fb083ife22c90acd76d461-at-mail.gmail.com} 2, 26 -- Date: Sat, 26 May 2007 15:28:06 -0400 2, 26 -- From: "Justin Kraft" {kraftpiano-at-gmail.com} 2, 26 -- To: microscopy-at-microscopy.com 2, 26 -- Subject: SEM: Vacuum level. 2, 26 -- MIME-Version: 1.0 2, 26 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 2, 26 -- Content-Transfer-Encoding: 7bit 2, 26 -- Content-Disposition: inline ==============================End of - Headers==============================
} Just out of curiosity, what should the vacuum level be to get } a steady electron beam in an older SEM? I grabbed a vacuum } gauge off of my sputter coater and measured the vacuum at 1.2 } x 10e-5. I don't think that's enough, though. Any thoughts?
10-5 would be good enough if we were speaking Pascals, and only if this vacuum could be assumed to be in the region of the gun.
cheerios :o) michael shaffer
SEM/MLA Research Coordinator {http://www.mun.ca/creait/maf/} Inco Innovation Centre Memorial University St. John's, Newfoundland
==============================Original Headers============================== 6, 21 -- From michael-at-shaffer.net Sat May 26 16:04:52 2007 6, 21 -- Received: from n007.sc1.he.tucows.com (smtpout1482.sc1.he.tucows.com [64.97.157.182]) 6, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4QL4qQv030848 6, 21 -- for {Microscopy-at-microscopy.com} ; Sat, 26 May 2007 16:04:52 -0500 6, 21 -- Received: from roamingwolf (205.251.83.78) by n007.sc1.he.tucows.com (7.2.069.1) (authenticated as michael-at-shaffer.net) 6, 21 -- id 465670BB0003CDD1; Sat, 26 May 2007 21:04:46 +0000 6, 21 -- From: "michael shaffer" {michael-at-shaffer.net} 6, 21 -- To: {kraftpiano-at-gmail.com} 6, 21 -- Cc: "MSA Microscopy list" {Microscopy-at-microscopy.com} 6, 21 -- References: {200705261928.l4QJSx6V018940-at-ns.microscopy.com} 6, 21 -- Subject: RE: [Microscopy] SEM: Vacuum level. 6, 21 -- Date: Sat, 26 May 2007 18:34:52 -0230 6, 21 -- Message-ID: {001001c79fd9$81d4e040$6401a8c0-at-CREAIT.MUN.CA} 6, 21 -- MIME-Version: 1.0 6, 21 -- Content-Type: text/plain; 6, 21 -- charset="US-ASCII" 6, 21 -- Content-Transfer-Encoding: 7bit 6, 21 -- X-Mailer: Microsoft Office Outlook 11 6, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 6, 21 -- In-Reply-To: {200705261928.l4QJSx6V018940-at-ns.microscopy.com} 6, 21 -- Thread-Index: Acefy43TpEbXUfKoRQqebBVPunDR0AADVwSw ==============================End of - Headers==============================
Sorry, I forgot to include the units in my vacuum measurement! I just committed the cardinal sin that I drill into my students- I wrote a number without a unit of measure!
Anyway, the vacuum measurement is 1.2x10e-5 torr, and is measured in the chamber itself. I can only assume that it is the same throughout the gun, as the sensor is in a location directly between the two locations, but on the vacuum system side.
--Justin.
On 5/26/07, Justin Kraft {kraftpiano-at-gmail.com} wrote: } Just out of curiosity, what should the vacuum level be to get a steady } electron beam in an older SEM? I grabbed a vacuum gauge off of my } sputter coater and measured the vacuum at 1.2 x 10e-5. I don't think } that's enough, though. Any thoughts? } } --Justin. }
==============================Original Headers============================== 4, 28 -- From kraftpiano-at-gmail.com Sat May 26 16:08:04 2007 4, 28 -- Received: from py-out-1112.google.com (py-out-1112.google.com [64.233.166.180]) 4, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4QL83Tx002815 4, 28 -- for {microscopy-at-microscopy.com} ; Sat, 26 May 2007 16:08:04 -0500 4, 28 -- Received: by py-out-1112.google.com with SMTP id d32so2126405pye 4, 28 -- for {microscopy-at-microscopy.com} ; Sat, 26 May 2007 14:08:03 -0700 (PDT) 4, 28 -- DKIM-Signature: a=rsa-sha1; c=relaxed/relaxed; 4, 28 -- d=gmail.com; s=beta; 4, 28 -- h=domainkey-signature:received:received:message-id:date:from:to:subject:in-reply-to:mime-version:content-type:content-transfer-encoding:content-disposition:references; 4, 28 -- b=CQU6/PO90XQWixSbTtwv62lpyJ94xNJ9/ursvRcRn6CD5Kf4ndwkdyO6dRiuGazBb4ZHHWlnnJRNGUqvNQH0p9gPzxUx8t6dKMHg3y/fv7s3mTKHIFeFdaflwsh4Wku6cW3Ya1Y/xk+/3Mkm7iKiKFZP/2GERkQfPtzzIl/iT5w= 4, 28 -- DomainKey-Signature: a=rsa-sha1; c=nofws; 4, 28 -- d=gmail.com; s=beta; 4, 28 -- h=received:message-id:date:from:to:subject:in-reply-to:mime-version:content-type:content-transfer-encoding:content-disposition:references; 4, 28 -- b=QeedACv0zP6uaGjA3hkDQC4AQIZ2rrUFqYCFOQqopKgdtIs73dldSWZ8pfeEwmTx3u2KP7R7x/ZOG3lacHdPsWWn322IetDP5egVLtmWj+pTnmiMutv2S3lBD5dUJJfiYHAvQr87awKWrA5ubzqNSHRT8/HlcTtYtZIMcz3mQ+c= 4, 28 -- Received: by 10.114.205.1 with SMTP id c1mr2098292wag.1180213683297; 4, 28 -- Sat, 26 May 2007 14:08:03 -0700 (PDT) 4, 28 -- Received: by 10.114.78.15 with HTTP; Sat, 26 May 2007 14:08:03 -0700 (PDT) 4, 28 -- Message-ID: {25e2b0d20705261408q3a7ff838k2f4d0b87a96c98d0-at-mail.gmail.com} 4, 28 -- Date: Sat, 26 May 2007 17:08:03 -0400 4, 28 -- From: "Justin Kraft" {kraftpiano-at-gmail.com} 4, 28 -- To: microscopy-at-microscopy.com 4, 28 -- Subject: Re: SEM: Vacuum level. 4, 28 -- In-Reply-To: {25e2b0d20705261228g395fb083ife22c90acd76d461-at-mail.gmail.com} 4, 28 -- MIME-Version: 1.0 4, 28 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 28 -- Content-Transfer-Encoding: 7bit 4, 28 -- Content-Disposition: inline 4, 28 -- References: {25e2b0d20705261228g395fb083ife22c90acd76d461-at-mail.gmail.com} ==============================End of - Headers==============================
} Anyway, the vacuum measurement is 1.2x10e-5 torr, and is } measured in the chamber itself. I can only assume that it is } the same throughout the gun, as the sensor is in a location } directly between the two locations, but on the vacuum system side.
I would consider this vacuum borderline, only because I've seen slightly better across a broad range of SEMs and probes that I have some experience with. Your vacuum might be considered average for the chamber region which is generally a large volume and usually subject to a small amount of outgassing. However, take the samples out and give the pumping 12 hours and 10-6 Torr should be achievable (e.g., my SEM achieves 1e-6 Torr easily.
However, your question is primarily about gun stability, and I might imagine a small leak in the region of the gun "could be" the reason for a relatively poor vacuum elsewhere. What exactly are your symptoms of instability, and what are your filament lifetimes? If instabilities aren't short term, abrupt and acute, and if your filament lifetimes are normal, I might believe the small leak isn't near the gun and that your problem can be lived with while you inspect, clean or replace o-rings in one region at a time.
Genuinely, Michael Shaffer :o)
SEM/MLA Research Coordinator http://www.mun.ca/creait/maf/
Inco Innovation Centre Memorial University St. John's, Newfoundland
} On 5/26/07, Justin Kraft {kraftpiano-at-gmail.com} wrote: } } Just out of curiosity, what should the vacuum level be to } get a steady } } electron beam in an older SEM? I grabbed a vacuum gauge off of my } } sputter coater and measured the vacuum at 1.2 x 10e-5. I } don't think } } that's enough, though. Any thoughts? } } } } --Justin.
==============================Original Headers============================== 11, 21 -- From michael-at-shaffer.net Sun May 27 09:04:48 2007 11, 21 -- Received: from n034.sc1.he.tucows.com (smtpout1490.sc1.he.tucows.com [64.97.157.190]) 11, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4RE4miR029723 11, 21 -- for {Microscopy-at-microscopy.com} ; Sun, 27 May 2007 09:04:48 -0500 11, 21 -- Received: from rarewolf (205.251.83.78) by n034.sc1.he.tucows.com (7.2.069.1) (authenticated as michael-at-shaffer.net) 11, 21 -- id 465895310000D344; Sun, 27 May 2007 14:04:43 +0000 11, 21 -- From: "michael shaffer" {michael-at-shaffer.net} 11, 21 -- To: {kraftpiano-at-gmail.com} 11, 21 -- Cc: "MSA Microscopy list" {Microscopy-at-microscopy.com} 11, 21 -- References: {200705262108.l4QL8OW2003987-at-ns.microscopy.com} 11, 21 -- Subject: RE: [Microscopy] Re: SEM: Vacuum level. 11, 21 -- Date: Sun, 27 May 2007 11:34:07 -0230 11, 21 -- Message-ID: {002001c7a067$e59ebe50$4701a8c0-at-rarewolf} 11, 21 -- MIME-Version: 1.0 11, 21 -- Content-Type: text/plain; 11, 21 -- charset="us-ascii" 11, 21 -- Content-Transfer-Encoding: 7bit 11, 21 -- X-Mailer: Microsoft Office Outlook 11 11, 21 -- Thread-Index: Acef2W90RdPVsEHOTeaH1xngB+RAzwAi7E/w 11, 21 -- In-Reply-To: {200705262108.l4QL8OW2003987-at-ns.microscopy.com} 11, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 ==============================End of - Headers==============================
I believe that most of us learned some time ago to disable color management in Photoshop for avoiding side-effects due to color space conversions. Indeed, most of us do not need CM enabled, until it's time to actually use it (e.g., accurate prints).
For those of you, like me, who end up with instrument image TIFs that are "indexed", you may have noticed rather extreme histogram shifts when you converted to "grayscale". This problem is eliminated by disabling CM; however, I've just noticed another problem even when CM is disabled. There is another option available under color settings only if you enable "more options". It is under "conversion options" and called "use dither (8bit/channel)", and used for randomizing 8bit quantization effects (usually a good thing). I never suspected this came into effect for indexed-to-grayscale conversions, but I can see significant changes to the histogram while the changes to the image are not significant at all.
This is not the end of it; i.e., I can still see small changes at the low end of the histogram even while CM and dithering are disabled. Has anyone found a remedy for this? That is, a simple way of stripping the LUT from an indexed TIF?
cheerios :o) michael shaffer
SEM/MLA Research Coordinator {http://www.mun.ca/creait/maf/} Inco Innovation Centre Memorial University St. John's, Newfoundland
==============================Original Headers============================== 6, 18 -- From michael-at-shaffer.net Sun May 27 12:52:02 2007 6, 18 -- Received: from n034.sc1.he.tucows.com (smtpout1460.sc1.he.tucows.com [64.97.157.160]) 6, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4RHq22t012194 6, 18 -- for {Microscopy-at-microscopy.com} ; Sun, 27 May 2007 12:52:02 -0500 6, 18 -- Received: from roamingwolf (205.251.83.78) by n034.sc1.he.tucows.com (7.2.069.1) (authenticated as michael-at-shaffer.net) 6, 18 -- id 4658953100010F6F for Microscopy-at-microscopy.com; Sun, 27 May 2007 17:52:02 +0000 6, 18 -- From: "michael shaffer" {michael-at-shaffer.net} 6, 18 -- To: "MSA Microscopy list" {Microscopy-at-microscopy.com} 6, 18 -- Subject: Yet another Photoshop pitfall (YAPP) 6, 18 -- Date: Sun, 27 May 2007 15:22:04 -0230 6, 18 -- Message-ID: {000e01c7a087$bf9fd2a0$6401a8c0-at-CREAIT.MUN.CA} 6, 18 -- MIME-Version: 1.0 6, 18 -- Content-Type: text/plain; 6, 18 -- charset="US-ASCII" 6, 18 -- Content-Transfer-Encoding: 7bit 6, 18 -- X-Mailer: Microsoft Office Outlook 11 6, 18 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 6, 18 -- Thread-Index: Acegh7w/BcYD7og3RBimMtX6b10fdg== ==============================End of - Headers==============================
While this is not a particularly "good" vacuum for your SEM as others have pointed out and you should likely look for leaks in order for that to improve, it should be fine for running your instrument.
I think, but I'll have to check, that you should be able to run your electron beam and image with an order of magnitude less vacuum (10-4 torr range).
Certainly, your instrument should be achieving better vacuum levels and you should look for small leaks. There could be out-gassing going on, there could be small leaks or any combination. Are you able to leave the instrument running under vacuum for a couple days and take the measurement at that point?
But if you are getting into the low 10-5 torr range, you should have no problem with beam stability or imaging. If you are, the cause is not your vacuum level...
dj
On Sat, 26 May 2007, kraftpiano-at-gmail.com wrote:
} Date: Sat, 26 May 2007 16:10:48 -0500 } From: kraftpiano-at-gmail.com } To: dljones-at-bestweb.net } Subject: [Microscopy] Re: SEM: Vacuum level. } } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Sorry, I forgot to include the units in my vacuum measurement! I just } committed the cardinal sin that I drill into my students- I wrote a } number without a unit of measure! } } Anyway, the vacuum measurement is 1.2x10e-5 torr, and is measured in } the chamber itself. I can only assume that it is the same throughout } the gun, as the sensor is in a location directly between the two } locations, but on the vacuum system side. } } --Justin. } } On 5/26/07, Justin Kraft {kraftpiano-at-gmail.com} wrote: } } Just out of curiosity, what should the vacuum level be to get a steady } } electron beam in an older SEM? I grabbed a vacuum gauge off of my } } sputter coater and measured the vacuum at 1.2 x 10e-5. I don't think } } that's enough, though. Any thoughts? } } } } --Justin. } } } } ==============================Original Headers============================== } 4, 28 -- From kraftpiano-at-gmail.com Sat May 26 16:08:04 2007 } 4, 28 -- Received: from py-out-1112.google.com (py-out-1112.google.com [64.233.166.180]) } 4, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4QL83Tx002815 } 4, 28 -- for {microscopy-at-microscopy.com} ; Sat, 26 May 2007 16:08:04 -0500 } 4, 28 -- Received: by py-out-1112.google.com with SMTP id d32so2126405pye } 4, 28 -- for {microscopy-at-microscopy.com} ; Sat, 26 May 2007 14:08:03 -0700 (PDT) } 4, 28 -- DKIM-Signature: a=rsa-sha1; c=relaxed/relaxed; } 4, 28 -- d=gmail.com; s=beta; } 4, 28 -- h=domainkey-signature:received:received:message-id:date:from:to:subject:in-reply-to:mime-version:content-type:content-transfer-encoding:content-disposition:references; } 4, 28 -- b=CQU6/PO90XQWixSbTtwv62lpyJ94xNJ9/ursvRcRn6CD5Kf4ndwkdyO6dRiuGazBb4ZHHWlnnJRNGUqvNQH0p9gPzxUx8t6dKMHg3y/fv7s3mTKHIFeFdaflwsh4Wku6cW3Ya1Y/xk+/3Mkm7iKiKFZP/2GERkQfPtzzIl/iT5w= } 4, 28 -- DomainKey-Signature: a=rsa-sha1; c=nofws; } 4, 28 -- d=gmail.com; s=beta; } 4, 28 -- h=received:message-id:date:from:to:subject:in-reply-to:mime-version:content-type:content-transfer-encoding:content-disposition:references; } 4, 28 -- b=QeedACv0zP6uaGjA3hkDQC4AQIZ2rrUFqYCFOQqopKgdtIs73dldSWZ8pfeEwmTx3u2KP7R7x/ZOG3lacHdPsWWn322IetDP5egVLtmWj+pTnmiMutv2S3lBD5dUJJfiYHAvQr87awKWrA5ubzqNSHRT8/HlcTtYtZIMcz3mQ+c= } 4, 28 -- Received: by 10.114.205.1 with SMTP id c1mr2098292wag.1180213683297; } 4, 28 -- Sat, 26 May 2007 14:08:03 -0700 (PDT) } 4, 28 -- Received: by 10.114.78.15 with HTTP; Sat, 26 May 2007 14:08:03 -0700 (PDT) } 4, 28 -- Message-ID: {25e2b0d20705261408q3a7ff838k2f4d0b87a96c98d0-at-mail.gmail.com} } 4, 28 -- Date: Sat, 26 May 2007 17:08:03 -0400 } 4, 28 -- From: "Justin Kraft" {kraftpiano-at-gmail.com} } 4, 28 -- To: microscopy-at-microscopy.com } 4, 28 -- Subject: Re: SEM: Vacuum level. } 4, 28 -- In-Reply-To: {25e2b0d20705261228g395fb083ife22c90acd76d461-at-mail.gmail.com} } 4, 28 -- MIME-Version: 1.0 } 4, 28 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed } 4, 28 -- Content-Transfer-Encoding: 7bit } 4, 28 -- Content-Disposition: inline } 4, 28 -- References: {25e2b0d20705261228g395fb083ife22c90acd76d461-at-mail.gmail.com} } ==============================End of - Headers============================== }
==============================Original Headers============================== 9, 21 -- From "dljones-at-bestweb.net"-at-bestweb.net Sun May 27 14:12:19 2007 9, 21 -- Received: from mta5.srv.hcvlny.cv.net (mta5.srv.hcvlny.cv.net [167.206.4.200]) 9, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4RJCIpQ024643 9, 21 -- for {microscopy-at-microscopy.com} ; Sun, 27 May 2007 14:12:19 -0500 9, 21 -- Received: from localhost (ool-44c62883.dyn.optonline.net [68.198.40.131]) 9, 21 -- by mta5.srv.hcvlny.cv.net 9, 21 -- (Sun Java System Messaging Server 6.2-6.01 (built Apr 3 2006)) 9, 21 -- with ESMTP id {0JIP002PGS0F9E90-at-mta5.srv.hcvlny.cv.net} for 9, 21 -- microscopy-at-microscopy.com; Sun, 27 May 2007 15:12:18 -0400 (EDT) 9, 21 -- Date: Sun, 27 May 2007 15:12:15 -0400 (Eastern Daylight Time) 9, 21 -- From: "David L. Jones" {"dljones-at-bestweb.net"-at-bestweb.net} 9, 21 -- Subject: Re: [Microscopy] Re: SEM: Vacuum level. 9, 21 -- In-reply-to: {200705262110.l4QLAm1P013185-at-ns.microscopy.com} 9, 21 -- X-X-Sender: dljones-at-pop-croton.bestweb.net 9, 21 -- To: kraftpiano-at-gmail.com 9, 21 -- Cc: microscopy-at-microscopy.com 9, 21 -- Message-id: {Pine.WNT.4.64.0705271501070.4024-at-dljtoshiba} 9, 21 -- MIME-version: 1.0 9, 21 -- Content-type: TEXT/PLAIN; charset=US-ASCII; format=flowed 9, 21 -- Content-transfer-encoding: 7BIT 9, 21 -- References: {200705262110.l4QLAm1P013185-at-ns.microscopy.com} ==============================End of - Headers==============================
That vacuum should be fine for a stable beam, provided, as Mike has said, that the vacuum in the gun is good. It would be worthwhile, IMHO, to make up a gauge-mounting flange right by the gun, there is a suitable place immediately above V3. A gauge near the gun is really useful for troubleshooting.
Do you still have any wds spectros mounted? Those things can leak a bit.
cheers
rtch
} } Just out of curiosity, what should the vacuum level be to get a steady } electron beam in an older SEM? I grabbed a vacuum gauge off of my } sputter coater and measured the vacuum at 1.2 x 10e-5. I don't think } that's enough, though. Any thoughts? } } --Justin. }
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
==============================Original Headers============================== 9, 27 -- From r.sims-at-auckland.ac.nz Sun May 27 15:54:58 2007 9, 27 -- Received: from mailhost.auckland.ac.nz (larry.its.auckland.ac.nz [130.216.10.122]) 9, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4RKsw9A005634 9, 27 -- for {microscopy-at-microscopy.com} ; Sun, 27 May 2007 15:54:58 -0500 9, 27 -- Received: from localhost (localhost.localdomain [127.0.0.1]) 9, 27 -- by mailhost.auckland.ac.nz (Postfix) with ESMTP id A92D0182A4; 9, 27 -- Mon, 28 May 2007 08:54:52 +1200 (NZST) 9, 27 -- X-Virus-Scanned: by amavisd-new at mailhost.auckland.ac.nz 9, 27 -- Received: from mailhost.auckland.ac.nz ([127.0.0.1]) 9, 27 -- by localhost (larry.its.auckland.ac.nz [127.0.0.1]) (amavisd-new, port 10024) 9, 27 -- with ESMTP id Ghh44pmN5yyn; Mon, 28 May 2007 08:54:52 +1200 (NZST) 9, 27 -- Received: from rs (r.sims.glg.auckland.ac.nz [130.216.59.18]) 9, 27 -- by mailhost.auckland.ac.nz (Postfix) with ESMTP id 83A46182A5; 9, 27 -- Mon, 28 May 2007 08:54:52 +1200 (NZST) 9, 27 -- From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} 9, 27 -- To: kraftpiano-at-gmail.com 9, 27 -- Date: Mon, 28 May 2007 08:55:55 +1200 9, 27 -- MIME-Version: 1.0 9, 27 -- Subject: Re: [Microscopy] SEM: Vacuum level. 9, 27 -- CC: microscopy-at-microscopy.com 9, 27 -- Message-ID: {465A991B.2777.A802B-at-localhost} 9, 27 -- Priority: normal 9, 27 -- In-reply-to: {200705261929.l4QJTq8Y019864-at-ns.microscopy.com} 9, 27 -- X-mailer: Pegasus Mail for Windows (4.21c) 9, 27 -- Content-type: text/plain; charset=US-ASCII 9, 27 -- Content-transfer-encoding: 7BIT 9, 27 -- Content-description: Mail message body ==============================End of - Headers==============================
I don't see the big deal about this. Zeiss TIFF images are indexed color. but the header contains specific info so the image files can be used with the separate TIFF editor.
I programmed F2 as a Photoshop action to convert the Zeiss indexed color files (8-bit and 16-bit) to 8-bit b/w and Save. It is really important to do this conversion since the indexed color images are quite worse than pure b/w.
No problem.
gary g.
At 09:53 AM 5/27/2007, you wrote:
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==============================Original Headers============================== 11, 20 -- From gary-at-gaugler.com Sun May 27 20:29:43 2007 11, 20 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 11, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l4S1Th4d023950 11, 20 -- for {microscopy-at-microscopy.com} ; Sun, 27 May 2007 20:29:43 -0500 11, 20 -- Message-Id: {200705280129.l4S1Th4d023950-at-ns.microscopy.com} 11, 20 -- Received: (qmail 4925 invoked from network); 27 May 2007 18:29:42 -0700 11, 20 -- Received: by simscan 1.1.0 ppid: 4922, pid: 4923, t: 0.1081s 11, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 11, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 11, 20 -- by qsmtp3 with SMTP; 27 May 2007 18:29:42 -0700 11, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.1.0.9 11, 20 -- Date: Sun, 27 May 2007 18:29:46 -0800 11, 20 -- To: michael-at-shaffer.net 11, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 11, 20 -- Subject: Re: [Microscopy] Yet another Photoshop pitfall (YAPP) 11, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 11, 20 -- In-Reply-To: {200705271753.l4RHrtcK015011-at-ns.microscopy.com} 11, 20 -- References: {200705271753.l4RHrtcK015011-at-ns.microscopy.com} 11, 20 -- Mime-Version: 1.0 11, 20 -- Content-Type: text/plain; charset=us-ascii; format=flowed; x-avg-checked=avg-ok-21395D19 ==============================End of - Headers==============================
Geoff McA asked for the reference to the paper I mentioned. It's Peters S et al; American Journal of Ophthalmology (2007) 143:995-1002; Ultrastructural findings in the primate eye after intravitreal injection of Bevacizumab. All it says is
"postfixed with 1% OsO4 at room temperature in 0.1 M cacodylate buffer (pH 7.4) for three hours, bloc-stained with uranyl acetate and lead citrate, and embedded in Epon after dehydration in a graded series of acetones"
I've Emailed asking for more details. I have the PDF if anyone would like to see the pics.
Diana
} ---------------------------------------------------------------------- } ------ } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } I've just read a paper where the author talks about block staining } with uranyl acetate and lead citrate. UAc yes, but Pbcitrate? Has } anyone heard of this? Does it work? The pictures in the paper were } very nice; good contrast and detail. With the recent talk about } general lack of contrast in specimens these days (I quite agree on } that; I find contrast poorer now than in the past), could this be } a new method? } } All opinions please! } } Cheers, } } Diana }
==============================Original Headers============================== 6, 18 -- From dianavd-at-eye.usyd.edu.au Sun May 27 22:03:44 2007 6, 18 -- Received: from galen.med.usyd.edu.au (nugalen.med.usyd.edu.au [129.78.36.39]) 6, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4S33iZi004234 6, 18 -- for {Microscopy-at-microscopy.com} ; Sun, 27 May 2007 22:03:44 -0500 6, 18 -- Received: from [129.78.203.144] (helo=[129.78.203.144]) 6, 18 -- by galen.med.usyd.edu.au with esmtp (Exim 4.43) 6, 18 -- id 1HsVWH-0007ML-W3; Mon, 28 May 2007 13:03:42 +1000 6, 18 -- Mime-Version: 1.0 (Apple Message framework v752.2) 6, 18 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 6, 18 -- Message-Id: {840EC60F-C8C0-4DD8-B2EF-76382CAACBB8-at-eye.usyd.edu.au} 6, 18 -- Cc: Geoff McAuliffe {mcauliff-at-umdnj.edu} 6, 18 -- Content-Transfer-Encoding: 7bit 6, 18 -- From: Diana van Driel {dianavd-at-eye.usyd.edu.au} 6, 18 -- Subject: lead citrate and block staining - more info 6, 18 -- Date: Mon, 28 May 2007 13:03:40 +1000 6, 18 -- To: Microscopy {Microscopy-at-microscopy.com} 6, 18 -- X-Mailer: Apple Mail (2.752.2) 6, 18 -- X-Spam-Score: -4.4 (----) ==============================End of - Headers==============================
The advice given by others is sound on measuring length and diameter (e.g. by area/length). Your fibres look very uniform in diameter though, so a lot of manual 'measure distance' clicks on a highly magnified live SEM image may be fine, and quicker, if your SEM PC can do it - save to Excel.
You can try binary 'erode' options on a thresholded fibre as well as skeletize (which can leave branches, but your fibres look ideal in this case). At Harwell Biomedical (now defunct) we used dedicated 'fibre' image analysis 'plug-ins' to measure length and diameter to help through-put (used with equally defunct software from Magiscan). We did a lot of work for the MMVF fibre manufacturers back then, and had sophisticated fibre activation techniques for in-vivo radio-tracing. I don't have access to my Image Pro (Bio-Rad LaserPix) key any more but look out for things like 'chord' and 'arc' length if offered for the skeletized line, and just make sure your length isn't the shortest distance [feret] between the fibre beginning and end co-ordinates. Do chat to your local Image Pro rep who will have almost certainly been asked this question before - www.mediacy.com. Not all programs are curvy fibre friendly, so you can sometimes use perimeter to roughly estimate length on virgin 'rectangular' curved fibres (divided by 2 and corrected for two diameters at the ends), unless the fibre surface has eroded.
Once you get a binary squiggly 'line' object on the overlay (say after skeletize), the fibre length measurement should be straightforward with any decent image analysis package. If you can't threshold the fibres correctly from the support background, try editing the thresholded binary image (exported to Photoshop if necessary as most modern image analysis packages seem the lack the obviously useful full binary editor - MetaMorph has a rather poor stab at it and promptly forgets your object editing for 'field' measurements). Editing images via a cheap Wacom Graphire tablet and stylus can be easier, but ImagePro did have the 'magnify' option to zoom into the image when using a mouse (or stylus). I notice that your fibres threshold well, but background 'particles' needed removing (probably simple with an object classifier removing say all objects below 5um in length or are more circular in shape (circularity)). Plus you will have to edit for length and possibly diameter measurements with overlapping fibres.
By the way, you must use the correct W.H.O. counting rules for counting fibres as they aren't as easy to score as rounder particles, e.g.
which is biased towards optical microscopy, but it's easily applied to SEM. Here you do need the low magnification of your original SEM image you showed us. Essentially you select random fields, and a countable fibre is a particle longer than 5ìm, with width less than 3ìm, and with a length:diameter ratio greater than 3:1 (for Health & Safety atmospheric monitoring - you probably know exactly which are your fibres). A countable fibre with both ends in the graticule area is counted as one fibre, a countable fibre with only one end in the area counts as half. A fibre completely crossing the graticule, with no ends in, is not counted. (An easy, practical method is to count the number of fibre ends in the graticule area and divide by two, having regard to the proper treatment of split fibres and clumps).... and so on. Fibre counting is always far quicker by eye, viewing captured images onscreen or down the microscope, and can be done live by SEM.
Just zoom in on random fibres or all those in the field counted to get your diameters (from 30+ fibres - we always did hundreds as our fibres were often recovered from lungs in in-vivo durability studies and fibre diameter could be highly variable). This generally done quickest while on the SEM, and we used a CamScan SEM with a full Quantimet image analysis software package on-line (it saves capturing loads of images, other than a few 'typical views') - and in those days our hard drives were in Mb. Naturally make sure all fields are chosen randomly to prevent bias.
I'll send another pdf that adds in a lot of useful fibre counting info (but check out the latest WHO (world health organisation) counting rules as I left the fibre toxicology field behind 8 years ago.
Regards
Keith
----------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Institute of Ophthalmology UCL, London EC1V 9EL
-----Original Message----- X-from: beaurega-at-westol.com [mailto:beaurega-at-westol.com] Sent: 25 May 2007 23:27 To: keith.morris-at-ucl.ac.uk
I agree with Jim after seeing the image.
A good rule of thumb in image analysis is to have about 30 features in each field of view that you are counting. However, some of that depends on the degree of dispersion (separation) of the features, their sizes, and in your case the aspect ratios. If you have a very good dispersion of smaller (rounder) features, then higher numbers of features can be counted per field but you risk flocculation affects.
Raise the mag to start with and count multiple fields of view.
Paul
At 02:00 PM 5/25/07 -0500, you wrote: } } } } --------------------------------------------------------------------------- - } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 5, 26 -- From beaurega-at-westol.com Fri May 25 17:22:44 2007 5, 26 -- Received: from smtp-gateway-2.winbeam.com (smtp-gateway-2.winbeam.com [64.84.97.67]) 5, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4PMMiDB012478 5, 26 -- for {microscopy-at-microscopy.com} ; Fri, 25 May 2007 17:22:44 -0500 5, 26 -- Received: from mail.winbeam.com (mail.winbeam.com [64.84.96.10]) 5, 26 -- by smtp-gateway-2.winbeam.com (8.13.1/8.12.8) with SMTP id l4PMJJsr014493 5, 26 -- for {microscopy-at-microscopy.com} ; Fri, 25 May 2007 18:19:20 -0400 5, 26 -- Received: (qmail 10736 invoked by uid 89); 25 May 2007 22:19:17 -0000 5, 26 -- Received: from pitts-69-72-109-13.dynamic-dialup.coretel.net (HELO beaurega) (69.72.109.13) 5, 26 -- by mail.winbeam.com with SMTP; 25 May 2007 22:19:17 -0000 5, 26 -- Message-Id: {3.0.6.32.20070525181922.007a08b0-at-pop3.norton.antivirus} 5, 26 -- X-Sender: beaurega/mail.westol.com-at-pop3.norton.antivirus 5, 26 -- X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.6 (32) 5, 26 -- Date: Fri, 25 May 2007 18:19:22 -0400 5, 26 -- To: jquinn-at-www.matscieng.sunysb.edu, microscopy-at-microscopy.com 5, 26 -- From: Beaurega {beaurega-at-westol.com} 5, 26 -- Subject: Re: [Microscopy] Re: AskAMicroscopist: Image Pro Analysis 5, 26 -- Question - URL with an Image 5, 26 -- In-Reply-To: {200705251900.l4PJ0den019188-at-ns.microscopy.com} 5, 26 -- Mime-Version: 1.0 5, 26 -- Content-Type: text/plain; charset="us-ascii" 5, 26 -- X--MailScanner-Information: - Please contact Technical Support for more information 5, 26 -- X--MailScanner: Found to be clean (courtesy of MailScanner) 5, 26 -- X--MailScanner-SpamCheck: not spam (whitelisted), SpamAssassin (not cached, 5, 26 -- timed out) 5, 26 -- X--MailScanner-From: beaurega-at-westol.com ==============================End of - Headers==============================
==============================Original Headers============================== 35, 22 -- From keith.morris-at-ucl.ac.uk Mon May 28 05:29:32 2007 35, 22 -- Received: from smtp4.global.net.uk (smtp4.global.net.uk [80.189.92.92]) 35, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4SATWSA023436 35, 22 -- for {microscopy-at-microscopy.com} ; Mon, 28 May 2007 05:29:32 -0500 35, 22 -- Received: from 5.27.125.91.gr6.adsl.brightview.com ([91.125.27.5] helo=loungepc) 35, 22 -- by smtp4.global.net.uk with esmtp (Exim 4.42) 35, 22 -- id 1HscTN-0003d3-T7; Mon, 28 May 2007 11:29:10 +0100 35, 22 -- From: "Keith Morris" {keith.morris-at-ucl.ac.uk} 35, 22 -- To: {vlk7-at-alfred.edu} 35, 22 -- Cc: {microscopy-at-microscopy.com} 35, 22 -- Subject: FW: [Microscopy] AskAMicroscopist: Image Pro Analysis 35, 22 -- Date: Mon, 28 May 2007 11:28:57 +0100 35, 22 -- Message-ID: {000301c7a113$00214760$0301a8c0-at-loungepc} 35, 22 -- MIME-Version: 1.0 35, 22 -- Content-Type: text/plain; 35, 22 -- charset="iso-8859-7" 35, 22 -- X-Mailer: Microsoft Office Outlook 11 35, 22 -- Thread-Index: AcefG9dkO1euyrTHT/WWca7VtC3SDQB5clUg 35, 22 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 35, 22 -- Authenticated-Sender: 35, 22 -- Content-Transfer-Encoding: 8bit 35, 22 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4SATWSA023436 ==============================End of - Headers==============================
I am sorry to pursue this question one more time, but WHY are you looking at the vacuum?
If the high voltage will switch on there is absolutely NO reason to look at the vacuum at that stage. Once the high voltage is operable if you then have discharge problems at high kV then you MAY have a leak. Lowering the kV the problem will be reduced but of course the strain on the high voltage system is also reduced so we do not necessarily prove it is a vacuum problem. It is almost impossible in the lab to check a high voltage cable as it may only fail at very high voltages, this is only a task for the experienced high voltage technician! The next phase sould be to look at filament life because it may surprise people to know that the instruments will run with a poor vacuum. Levels inferior to five times ten minus four torr have been quite normal in SEM and TEM in the past without high voltage discharge problems.
With over 40 years experience with the problems that beset TEM, SEM and EDS one of the major problems we all suffer from is pre judging a fault. When a fault occurs the question should be "what did you do last?". I have discussed this problem with Justin over a few weeks and I fail to see that the vacuum is at fault. The problem seems to me to be a high voltage cable breaking down.
There are ways of checking this out and finding a reduced cost fix rather than going to the original equipment manufacturer!
Steve Chapman Senior Consultant Protrain For electron microscopy consultancy and training world wide Tel +44 1280 816512 Fax +44 1280 814007 Mobile +44 7711 606967 Web www.emcourses.com
----- Original Message ----- X-from: {kraftpiano-at-gmail.com} To: {protrain-at-emcourses.com} Sent: Saturday, May 26, 2007 8:29 PM
Actually, the reason I was thinking it was the vacuum is exactly the "What happened last" question- there was a major leak, and in the process of repairing it, I think I may have lost some of the diffusion pump oil because it wasn't fully cooled down when I opened up the valve head above it to replace the O-ring. Then, the O-ring that I got was slightly larger than the O-ring that I removed. Other than that, nothing has changed a bit. Everything else was working perfectly until the aforementioned vacuum leak. I also discovered (In the process of troubleshooting things yesterday) that the vacuum gauge on the SEM is wildly off base. Late last night I was able to get an absolute vacuum reading from a gauge (Rather than convert the 0-200 scale on the built-in gauge) and the vacuum is actually only at about 120 millitorr, not the 1.2x10e-5 I stated earlier (That number was gained by a false conversion factor between the built-in gauge and reality).
I'm still not seeing, however, how removing the high voltage cable shows that it is the cable breaking down. The HV switches on, and no emission is measured. When the cable is plugged in, the HV switches on, and emission is measured at 20-30 (Out of 200, using the same arbitrary meter that the vacuum uses) then slowly dies down again. As to it not switching on if the vacuum level is low, I discovered that the high voltage will switch on as long as the vacuum meter registers high enough, which doesn't necessarily have anything to do with the actual vacuum, as I found out at about 1:00 AM this morning.
It seems that since the emission current is measured by how much current is actually flowing, removing the HV cable just proves that no current is flowing at all without it, and with it some flows, but then something breaks down. I'm not sure how that limits it to the HV cable.
I'm sorry if it seems like I'm just giving out information that should have been given out a while ago, (Or cancels out previous information) but I just got the vacuum gauge setup properly late last night.
--Justin.
On 5/28/07, protrain-at-emcourses.com {protrain-at-emcourses.com} wrote: } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Hi Again } } I am sorry to pursue this question one more time, but WHY are you looking at } the vacuum? } } If the high voltage will switch on there is absolutely NO reason to look at } the vacuum at that stage. } Once the high voltage is operable if you then have discharge problems at } high kV then you MAY have a leak. Lowering the kV the problem will be } reduced but of course the strain on the high voltage system is also reduced } so we do not necessarily prove it is a vacuum problem. It is almost } impossible in the lab to check a high voltage cable as it may only fail at } very high voltages, this is only a task for the experienced high voltage } technician! The next phase sould be to look at filament life because it may } surprise people to know that the instruments will run with a poor vacuum. } Levels inferior to five times ten minus four torr have been quite normal in } SEM and TEM in the past without high voltage discharge problems. } } With over 40 years experience with the problems that beset TEM, SEM and EDS } one of the major problems we all suffer from is pre judging a fault. When a } fault occurs the question should be "what did you do last?". I have } discussed this problem with Justin over a few weeks and I fail to see that } the vacuum is at fault. The problem seems to me to be a high voltage cable } breaking down. } } There are ways of checking this out and finding a reduced cost fix rather } than going to the original equipment manufacturer! } } Steve Chapman } Senior Consultant Protrain } For electron microscopy consultancy and training world wide } Tel +44 1280 816512 Fax +44 1280 814007 } Mobile +44 7711 606967 Web www.emcourses.com } } } ----- Original Message ----- } X-from: {kraftpiano-at-gmail.com} } To: {protrain-at-emcourses.com} } Sent: Saturday, May 26, 2007 8:29 PM } Subject: [Microscopy] SEM: Vacuum level. } } } } } } } } } } ---------------------------------------------------------------------------- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------------- } } } } Just out of curiosity, what should the vacuum level be to get a steady } } electron beam in an older SEM? I grabbed a vacuum gauge off of my } } sputter coater and measured the vacuum at 1.2 x 10e-5. I don't think } } that's enough, though. Any thoughts? } } } } --Justin. } } } } ==============================Original } } Headers============================== } } 2, 26 -- From kraftpiano-at-gmail.com Sat May 26 14:28:06 2007 } } 2, 26 -- Received: from wa-out-1112.google.com (wa-out-1112.google.com } } [209.85.146.182]) } } 2, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } } l4QJS6sc018284 } } 2, 26 -- for {microscopy-at-microscopy.com} ; Sat, 26 May 2007 14:28:06 -0500 } } 2, 26 -- Received: by wa-out-1112.google.com with SMTP id j5so466378wah } } 2, 26 -- for {microscopy-at-microscopy.com} ; Sat, 26 May 2007 } } 12:28:06 -0700 (PDT) } } 2, 26 -- DKIM-Signature: a=rsa-sha1; c=relaxed/relaxed; } } 2, 26 -- d=gmail.com; s=beta; } } 2, 26 -- } } h=domainkey-signature:received:received:message-id:date:from:to:subject:mime-version:content-type:content-transfer-encoding:content-disposition; } } 2, 26 -- } } b=mF1yFAt4zQgEOCJKeNzwygekp8ZE/db54wXobsSKwofa2CazIlAwgbw25uvo8xGCbiB3/nNQSFxHzx4K2HBERScwHErSxVYTAj0Kq4Iex4H8VA6Dz52XgdYnMyea82zokTmhgON7TZTSJjI+iFzXsmxuXgm4ULI+Oyai7gHu+Jk= } } 2, 26 -- DomainKey-Signature: a=rsa-sha1; c=nofws; } } 2, 26 -- d=gmail.com; s=beta; } } 2, 26 -- } } h=received:message-id:date:from:to:subject:mime-version:content-type:content-transfer-encoding:content-disposition; } } 2, 26 -- } } b=g2BLT42OeNHc/SIs4cL4WfBilfLyzgUf6maiRBhhc3w5bSovMX9arWLdkVH6inU4QgO6jPiSCiwWkE/R5Lq0c8VuBBVAWsPShwHhcdm49/jJOaBQfSHJesKY04Xn+8UjGp7X6mpmr+VkdSUMXW5vdFFC/gqvhue/32AkzV6bFKs= } } 2, 26 -- Received: by 10.114.14.1 with SMTP id } } 1mr2063238wan.1180207686080; } } 2, 26 -- Sat, 26 May 2007 12:28:06 -0700 (PDT) } } 2, 26 -- Received: by 10.114.78.15 with HTTP; Sat, 26 May 2007 } } 12:28:06 -0700 (PDT) } } 2, 26 -- Message-ID: } } {25e2b0d20705261228g395fb083ife22c90acd76d461-at-mail.gmail.com} } } 2, 26 -- Date: Sat, 26 May 2007 15:28:06 -0400 } } 2, 26 -- From: "Justin Kraft" {kraftpiano-at-gmail.com} } } 2, 26 -- To: microscopy-at-microscopy.com } } 2, 26 -- Subject: SEM: Vacuum level. } } 2, 26 -- MIME-Version: 1.0 } } 2, 26 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed } } 2, 26 -- Content-Transfer-Encoding: 7bit } } 2, 26 -- Content-Disposition: inline } } ==============================End of - } } Headers============================== } } } } } ==============================Original Headers============================== } 11, 27 -- From protrain-at-emcourses.com Mon May 28 05:41:20 2007 } 11, 27 -- Received: from smtp1.pri.skybb.uk.easynet.net (smtp1.pri.skybb.uk.easynet.net [87.86.189.69]) } 11, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4SAfJcm002620 } 11, 27 -- for {microscopy-at-microscopy.com} ; Mon, 28 May 2007 05:41:19 -0500 } 11, 27 -- Received: from [90.194.114.199] (helo=advent) } 11, 27 -- by smtp1.pri.skybb.uk.easynet.net with smtp (Exim 4.62) } 11, 27 -- (envelope-from {protrain-at-emcourses.com} ) } 11, 27 -- id 1Hscf6-0003bP-B7; Mon, 28 May 2007 11:41:16 +0100 } 11, 27 -- Message-ID: {005d01c7a114$e7a708d0$0200a8c0-at-advent} } 11, 27 -- Reply-To: "Steve Chapman" {protrain-at-emcourses.com} } 11, 27 -- From: "Steve Chapman" {protrain-at-emcourses.com} } 11, 27 -- To: "Justin Kraft" {kraftpiano-at-gmail.com} } 11, 27 -- Cc: "American Soc" {microscopy-at-microscopy.com} } 11, 27 -- References: {200705261929.l4QJTv4i019951-at-ns.microscopy.com} } 11, 27 -- Subject: Re: [Microscopy] SEM: Vacuum level. } 11, 27 -- Date: Mon, 28 May 2007 11:39:00 +0100 } 11, 27 -- Organization: Protrain } 11, 27 -- MIME-Version: 1.0 } 11, 27 -- Content-Type: text/plain; } 11, 27 -- format=flowed; } 11, 27 -- charset="iso-8859-1"; } 11, 27 -- reply-type=original } 11, 27 -- Content-Transfer-Encoding: 7bit } 11, 27 -- X-Priority: 3 } 11, 27 -- X-MSMail-Priority: Normal } 11, 27 -- X-Mailer: Microsoft Outlook Express 6.00.2900.3028 } 11, 27 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 } ==============================End of - Headers============================== }
==============================Original Headers============================== 6, 28 -- From kraftpiano-at-gmail.com Mon May 28 08:07:17 2007 6, 28 -- Received: from wr-out-0506.google.com (wr-out-0506.google.com [64.233.184.236]) 6, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4SD7Gh7017447 6, 28 -- for {microscopy-at-microscopy.com} ; Mon, 28 May 2007 08:07:16 -0500 6, 28 -- Received: by wr-out-0506.google.com with SMTP id 50so637130wra 6, 28 -- for {microscopy-at-microscopy.com} ; Mon, 28 May 2007 06:07:16 -0700 (PDT) 6, 28 -- DKIM-Signature: a=rsa-sha1; c=relaxed/relaxed; 6, 28 -- d=gmail.com; s=beta; 6, 28 -- h=domainkey-signature:received:received:message-id:date:from:to:subject:in-reply-to:mime-version:content-type:content-transfer-encoding:content-disposition:references; 6, 28 -- b=MYZWBOUWjjVCiDplafg+2a26QuwCaNJhpYzG4HMwAYpvO5xQErnxS8RofpHj9rWcZcHzs3d8LnUncNJSs2mYIPBeOg7gYTKRLjN1/CRKZ9NrqR4LDU9vxKGqY1lHvYJyl5DZd1LCDRFtZRrCXGlQsKTun5r4BOYkRCOcs1N0tKc= 6, 28 -- DomainKey-Signature: a=rsa-sha1; c=nofws; 6, 28 -- d=gmail.com; s=beta; 6, 28 -- h=received:message-id:date:from:to:subject:in-reply-to:mime-version:content-type:content-transfer-encoding:content-disposition:references; 6, 28 -- b=XB5B8MEuJQyo8nU8WaUpsn+GFUelhaUw1knWEj6A/4dJfTf1p1otXSgLR3TzmKFAJ3MbLrbkiwRsAKvEuIQGY/WtvpRHsUhe2PDULRrnUiNkVoU4p7+EKSwjnUFsCFedXnebj83iNW1Sw2oXcp5Qu9hssfv4EPaqGKxrwtDUKds= 6, 28 -- Received: by 10.114.177.1 with SMTP id z1mr2936536wae.1180357635050; 6, 28 -- Mon, 28 May 2007 06:07:15 -0700 (PDT) 6, 28 -- Received: by 10.114.78.15 with HTTP; Mon, 28 May 2007 06:07:14 -0700 (PDT) 6, 28 -- Message-ID: {25e2b0d20705280607j2e583995s4c818633e7579bfa-at-mail.gmail.com} 6, 28 -- Date: Mon, 28 May 2007 09:07:14 -0400 6, 28 -- From: "Justin Kraft" {kraftpiano-at-gmail.com} 6, 28 -- To: protrain-at-emcourses.com, microscopy-at-microscopy.com 6, 28 -- Subject: Re: [Microscopy] Re: SEM: Vacuum level. 6, 28 -- In-Reply-To: {200705281045.l4SAjtWv013337-at-ns.microscopy.com} 6, 28 -- MIME-Version: 1.0 6, 28 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 6, 28 -- Content-Transfer-Encoding: 7bit 6, 28 -- Content-Disposition: inline 6, 28 -- References: {200705281045.l4SAjtWv013337-at-ns.microscopy.com} ==============================End of - Headers==============================
The crunch here is not what is the vacuum BUT, can you get a beam?
I have run Electron Guns under a good rotary pump pressure, if the vacuum is not good enough the high voltage will short out to the walls of the system and you will hear high voltage discharge in the gun. Forget the vacuum and try to get a beam. When you draw a current, if the cable is at fault, the system will breakdown, currents will fall to zero! Only by trying to obtain a beam will you find out what is going on in your gun. You say that with the cable removed you do not obtain a reading, good no breakdown. You then say with the cable fitted you obtain a reading which falls to zero, is this the natural surge or a sign that the cable has broken down? Now if you try to heat the filament can you obtain an emission reading that will hold and then find the beam?
Just forget about the vacuum and run the gun. If the gun will not work you have proven that the high voltage system is the problem and a check on the cable in my mind will prove that the cable is at fault.
Steve Chapman Senior Consultant Protrain For electron microscopy consultancy and training world wide Tel +44 1280 816512 Fax +44 1280 814007 Mobile +44 7711 606967 Web www.emcourses.com
----- Original Message ----- X-from: "Justin Kraft" {kraftpiano-at-gmail.com} To: {protrain-at-emcourses.com} ; {microscopy-at-microscopy.com} Sent: Monday, May 28, 2007 2:07 PM
Diana,
David Elliott kindly emailed me his Microscopy Today article and protocol at my request. After aqueous Glut-OsO4 fix and UrAc bloc stain,and EtOH dehydration, he uses the lead as Pb acetate, saturated, in 50:50 EtOH:Acetone, then shifts into acetone to perform epoxy resin infiltration.
In the ref you cited, Dr Peters uses Pb Citrate, apparently aqueous, in a UrAc --} PbCit sequence before acetone dehydration.
So there seems a little leeway as to which lead salt and what stage to apply it, but only acetone reported as the transition solvent to epoxy resin.
The staining looks fine in both papers. I'll be trying it to compare with the contrast levels we get using Tannic Acid in the primary fix, and KMnO4-SatoPb as section stain sequence.
(I was clearly mistaken in my off-list warnings to you about curing and cutting problems I thought likely with the double bloc stain.)
-mike reedy-
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Geoff McA asked for the reference to the paper I mentioned. It's Peters S et al; American Journal of Ophthalmology (2007) 143:995-1002; Ultrastructural findings in the primate eye after intravitreal injection of Bevacizumab. All it says is
"postfixed with 1% OsO4 at room temperature in 0.1 M cacodylate buffer (pH 7.4) for three hours, bloc-stained with uranyl acetate and lead citrate, and embedded in Epon after dehydration in a graded series of acetones"
I've Emailed asking for more details. I have the PDF if anyone would like to see the pics.
Diana
} ---------------------------------------------------------------------- } ------ } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } I've just read a paper where the author talks about block staining } with uranyl acetate and lead citrate. UAc yes, but Pbcitrate? Has } anyone heard of this? Does it work? The pictures in the paper were } very nice; good contrast and detail. With the recent talk about } general lack of contrast in specimens these days (I quite agree on } that; I find contrast poorer now than in the past), could this be } a new method? } } All opinions please! } } Cheers, } } Diana }
==============================Original Headers============================== 6, 18 -- From dianavd-at-eye.usyd.edu.au Sun May 27 22:03:44 2007 6, 18 -- Received: from galen.med.usyd.edu.au (nugalen.med.usyd.edu.au [129.78.36.39]) 6, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4S33iZi004234 6, 18 -- for {Microscopy-at-microscopy.com} ; Sun, 27 May 2007 22:03:44 -0500 6, 18 -- Received: from [129.78.203.144] (helo=[129.78.203.144]) 6, 18 -- by galen.med.usyd.edu.au with esmtp (Exim 4.43) 6, 18 -- id 1HsVWH-0007ML-W3; Mon, 28 May 2007 13:03:42 +1000 6, 18 -- Mime-Version: 1.0 (Apple Message framework v752.2) 6, 18 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 6, 18 -- Message-Id: {840EC60F-C8C0-4DD8-B2EF-76382CAACBB8-at-eye.usyd.edu.au} 6, 18 -- Cc: Geoff McAuliffe {mcauliff-at-umdnj.edu} 6, 18 -- Content-Transfer-Encoding: 7bit 6, 18 -- From: Diana van Driel {dianavd-at-eye.usyd.edu.au} 6, 18 -- Subject: lead citrate and block staining - more info 6, 18 -- Date: Mon, 28 May 2007 13:03:40 +1000 6, 18 -- To: Microscopy {Microscopy-at-microscopy.com} 6, 18 -- X-Mailer: Apple Mail (2.752.2) 6, 18 -- X-Spam-Score: -4.4 (----) ==============================End of - Headers==============================
-- -mike reedy-
************************ Michael K. Reedy, M.D. Duke Univ. Med. Center Dept. Cell Biology, Box 3011 (for U.S. Mail) 458 Alex Sands Bldg, Research Dr (courier) Durham, NC 27710
We have a Philips EM300, TEM with various user, service & installation manuals plus some accessories, including single-tilt goniometer available.
This instrument was initially installed in the University of Victoria (Biology), BC Canada in the early 1970's.
It was moved to Simon Fraser University (Physics), sometime in the 1990's, and was acquired by Trinity Western (Biology) in 2000.
This instrument was demonstrated to be functional in its TWU location. Actual use has been extremely light.
We are interested in offering the instrument "free - as is where is". If assistance in disassembly/packing/shipping is required, those costs will be negotiated.
Please direct your inquiries to me, as convenient.
Darcy
Mr. Darcy Kehler, B.Sc. Sr. Lab Supervisor, Biology Faculty of Natural and Applied Sciences Trinity Western University p: 604.888.7511 ext. 3249 F: 604.513.2018 Langley, BC, Canada, V2Y 1Y1
==============================Original Headers============================== 9, 26 -- From kehler-at-twu.ca Mon May 28 17:11:21 2007 9, 26 -- Received: from lnxms2.twu.ca (lnxms2.twu.ca [64.114.134.150]) 9, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4SMBLwf011288 9, 26 -- for {Microscopy-at-microscopy.com} ; Mon, 28 May 2007 17:11:21 -0500 9, 26 -- Received: from lnxms2.twu.ca (lnxms2.twu.ca [127.0.0.1]) 9, 26 -- by localhost (Postfix) with SMTP id 3C9FA15801F 9, 26 -- for {Microscopy-at-microscopy.com} ; Mon, 28 May 2007 15:14:44 -0700 (PDT) 9, 26 -- Received: from ES1.twu.ca (es1.twu.ca [10.10.118.48]) 9, 26 -- by lnxms2.twu.ca (Postfix) with ESMTP id 3185A158006 9, 26 -- for {Microscopy-at-microscopy.com} ; Mon, 28 May 2007 15:14:44 -0700 (PDT) 9, 26 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 9, 26 -- Content-class: urn:content-classes:message 9, 26 -- MIME-Version: 1.0 9, 26 -- Content-Type: text/plain; 9, 26 -- charset="us-ascii" 9, 26 -- Subject: TEM available 9, 26 -- Date: Mon, 28 May 2007 15:14:55 -0700 9, 26 -- Message-ID: {A1A197BB3A238F408EA59B45A03EDC6803F78C2E-at-ES1.twu.ca} 9, 26 -- X-MS-Has-Attach: 9, 26 -- X-MS-TNEF-Correlator: 9, 26 -- Thread-Topic: TEM available 9, 26 -- thread-index: AcehdZ8tb8tWGQrBQuWXJuOr/RgUTg== 9, 26 -- From: "Darcy Kehler" {kehler-at-twu.ca} 9, 26 -- To: {Microscopy-at-microscopy.com} 9, 26 -- Content-Transfer-Encoding: 8bit 9, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4SMBLwf011288 ==============================End of - Headers==============================
The beam is orthogonal only if the specimen is e.g. perfectly flat on the specimen stub and the stub is exactly at zero degrees tilt. Like looking down on a pencil on your desktop. Push down on the point of the pencil and the opposite end pops up. Still looking *straight down* on the pencil, the image of the pencil will now be shorter. Try the shadow trick to see what I mean. The image on the view CRT, or as collected by the imaging system, is the 2D projection of the 3D image. And, the beam is only orthogonal at the exact center of the scan raster. Anywhere else, and it is approaching the specimen at some angle.
_/___ versus __|__
This is only important on perfectly flat specimens, though. On most normal specimens, and anything biological, the surface topography means the beam is very rarely perfectly orthogonal to the specimen surface. And, it may not affect the signal enough to matter. So we ignore the effect. For high-precision metrology or imaging of things like single molecules, I suspect this effect could become important.
Phil
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==============================Original Headers============================== 6, 22 -- From oshel1pe-at-cmich.edu Tue May 29 07:32:21 2007 6, 22 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 6, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4TCWLb3011146 6, 22 -- for {Microscopy-at-microscopy.com} ; Tue, 29 May 2007 07:32:21 -0500 6, 22 -- Received: from egatea.central.cmich.local ([141.209.15.74]) 6, 22 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id l4TCsjrG029099 6, 22 -- for {Microscopy-at-microscopy.com} ; Tue, 29 May 2007 08:55:12 -0400 6, 22 -- Received: from [141.209.160.249] ([141.209.160.249]) by egatea.central.cmich.local with Microsoft SMTPSVC(6.0.3790.1830); 6, 22 -- Tue, 29 May 2007 08:32:11 -0400 6, 22 -- Mime-Version: 1.0 6, 22 -- Message-Id: {f06230902c281cc618935-at-[141.209.160.249]} 6, 22 -- In-Reply-To: {200705260652.l4Q6q8xT017561-at-ns.microscopy.com} 6, 22 -- References: {200705260652.l4Q6q8xT017561-at-ns.microscopy.com} 6, 22 -- Date: Tue, 29 May 2007 08:32:10 -0400 6, 22 -- To: Microscopy-at-microscopy.com 6, 22 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 6, 22 -- Subject: [Microscopy] Re: SEM beam-specimen relations (subject line change) 6, 22 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 6, 22 -- X-OriginalArrivalTime: 29 May 2007 12:32:11.0384 (UTC) FILETIME=[6139AF80:01C7A1ED] 6, 22 -- X-CanItPRO-Stream: default 6, 22 -- X-Spam-Score: -4 () L_EXCH_MF 6, 22 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
The submission date for abstracts (in a two-page format, very similar to the M and M format) has been changed to June 30, 2007. There is still a month to get your paper in to this exciting meeting.
9th InterAmerican Congress on Electron Microscopy
Cusco Peru
September 23 - 28, 2007
One of the major electron microscopy meetings this year will be held in the heart of the Inca empire. Machu Picchu, one of the world's most famous and wondrous places is nearby.
Leading microscopists from across the Americas (and from the rest of the world) will be presenting their latest work.
Details of how to register (which must be done before the abstract is submitted) and of how to submit the abstract, can be found at the web site: http://www.ciasem2007.com/ (To change to English click the little blue button to the right.)
-- ........... 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
==============================Original Headers============================== 11, 21 -- From jae5-at-lehigh.edu Tue May 29 08:19:35 2007 11, 21 -- Received: from rain.CC.Lehigh.EDU (rain.CC.Lehigh.EDU [128.180.39.20]) 11, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4TDJY4c023372 11, 21 -- for {Microscopy-at-microscopy.com} ; Tue, 29 May 2007 08:19:34 -0500 11, 21 -- Received: from [127.0.0.1] (Dyn055141.mat.Lehigh.EDU [128.180.55.141]) 11, 21 -- (authenticated bits=0) 11, 21 -- by rain.CC.Lehigh.EDU (8.14.1/8.14.1) with ESMTP id l4TDJXRZ018715 11, 21 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NOT) 11, 21 -- for {Microscopy-at-microscopy.com} ; Tue, 29 May 2007 09:19:33 -0400 11, 21 -- Message-ID: {465C2866.2090105-at-lehigh.edu} 11, 21 -- Date: Tue, 29 May 2007 09:19:34 -0400 11, 21 -- From: Alwyn Eades {jae5-at-lehigh.edu} 11, 21 -- Organization: Lehigh University 11, 21 -- User-Agent: Thunderbird 1.5.0.10 (Windows/20070221) 11, 21 -- MIME-Version: 1.0 11, 21 -- To: Microscopy-at-microscopy.com 11, 21 -- Subject: Changed submission date CIASEM 2007 Cusco 11, 21 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 11, 21 -- Content-Transfer-Encoding: 7bit 11, 21 -- X-Virus-Scanned: ClamAV 0.90.2/3323/Tue May 29 08:10:43 2007 on rain.CC.Lehigh.EDU 11, 21 -- X-Virus-Status: Clean ==============================End of - Headers==============================
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Email: chunmin.wang-at-ingeniapolymers.com Name: Chunmin Wang
I just got one leitz 1350 heating stage from ebay and I want to connect it to our lab microscope. However, I cannot find the right temperature controller for the stage. Can anyone who has the same stage kindly tell me the model of the controller? Thank you,
Chunmin Wang
Research Engineer Ingenia Polymers Corp. 565 Greenwich Street Brantford, Ontario N3T 5M8 CANADA Tel: 519-758-8941 ext: 1020 Fax: 519-758-1254
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Title-Subject: [Filtered] 2 OPEN POSITIONS- FIB Applications and La Manager
Question: Due to growth, Carl Zeiss SMT Inc. is seeking two experienced FIB Product Managers, to be located at our new headquarters in Peabody, MA.
Responsibilities include overall FIB product management-related sales support activities, specifically instrument demonstrations, sample processing, prospect presentations, preparation and development of promotional items and sales tools, and customer training. Responsibilities also include overall FIB applications expertise in order to assist the Companyís sales efforts in achieving our instrument orders and sales targets, to equip the Companyís prospects and customers with in-depth product knowledge and capabilities enabling them to achieve high levels of operator satisfaction, to ensure the optimum product performance of our FIB instruments, and to assist the Companyís technical service efforts by disseminating appropriate product information and providing technical support. An advanced degree in a related field and extensive prior experience with FIBs in a corporate setting is required. One position requires frequent travel (both domestic and international to our facilities in Germany), a customer-focused mentality, and excellent communication skills. The Lab Manager position requires successful prior experience managing a lab. Compensation is commensurate with experience and Carl Zeiss SMT Inc. provides a comprehensive benefits package. An Affirmative Action Employer M/F/D/V Contact Beth Bressan, bressan-at-smt.zeiss.com All replies are confidential.
I blotted a suspension of particles in a salt solution on a nitrocellulose membrane (those used for protein blotting, 0,2 µm pore size) and washed it extensively under vacuum to remove the salts. Then I dried the membrane, glued it on a alu stub and carbon coated it (5 sec at 4 V). Now when I observe in SEM at 12 kV the image moves a lot, especially the luminosity. The region around the scanned area (I see it when I zoom out) is saturated (white). I suspect it is charged. In EDX the sensitivity is very low, much lower than expected. This is specific for this sample, other samples work good in EDX. I tried to observe in low vacuum mode (10 Pa) but at 12 kV I can see nothing. At 20 kV I get an image although the luminosity still is very unstable and the quality is far worse than in high vacuum mode. However it is possible to get an EDX spectrum in these conditions.
I was very surprised the way this membrane works in SEM and particularly in EDX. I already analyzed the same suspension directly deposited on a carbon tabs and I had good conditions to work.
1) Should I expected heavy loading in such a membrane, even carbon coated? 2) Does (charge) loading of the sample hamper the EDX analysis? 3) What would be a solution? coat more with carbon? I would like to avoid gold since it has a band that superimposes on a band of interest in EDX and could absorb some signal.
Stephane
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==============================Original Headers============================== 8, 19 -- From nizets2-at-yahoo.com Wed May 30 03:54:05 2007 8, 19 -- Received: from web37413.mail.mud.yahoo.com (web37413.mail.mud.yahoo.com [209.191.91.145]) 8, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l4U8s5sU027189 8, 19 -- for {microscopy-at-microscopy.com} ; Wed, 30 May 2007 03:54:05 -0500 8, 19 -- Received: (qmail 10486 invoked by uid 60001); 30 May 2007 08:54:05 -0000 8, 19 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 8, 19 -- s=s1024; d=yahoo.com; 8, 19 -- h=X-YMail-OSG:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding:Message-ID; 8, 19 -- b=xLcKvefYBL3FzasQ1WkA66brNXZSpRI1FQ+MXNWpdVsxwwll2w3VbdscCVGN6IgFAKEQDJphrBlG+OeQUzXJ1rewC6pVpg13XfUtjh78sJpVZ5Rq4umzKlZ6I2odQ7upgv6uuTK0wmm7UQPa13eb1NTYe6xJmV547a/p74aMk8I=; 8, 19 -- X-YMail-OSG: m_Xni.YVM1kcok4fajUssR0t01He.lBHc4HvTH8fgEpwUWumfHB3MV3Vo5xgIh0cDDcgCM3kKxXSD.59BQakGob.UpjKMixd8tgEha3ThVMnd4ONlAflH3J0JUFDiA-- 8, 19 -- Received: from [80.122.101.102] by web37413.mail.mud.yahoo.com via HTTP; Wed, 30 May 2007 01:54:05 PDT 8, 19 -- Date: Wed, 30 May 2007 01:54:05 -0700 (PDT) 8, 19 -- From: Stephane Nizet {nizets2-at-yahoo.com} 8, 19 -- Subject: Nitrocellulose membranes and SEM 8, 19 -- To: microscopy-at-microscopy.com 8, 19 -- MIME-Version: 1.0 8, 19 -- Content-Type: text/plain; charset=iso-8859-1 8, 19 -- Content-Transfer-Encoding: 8bit 8, 19 -- Message-ID: {155425.8726.qm-at-web37413.mail.mud.yahoo.com} ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dls_douglasl-at-yahoo.com.br) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, May 29, 2007 at 00:41:27 ---------------------------------------------------------------------------
Email: dls_douglasl-at-yahoo.com.br Name: Douglas da Silva
Organization: Universidade federal do Rio Grande do Sul
Education: Graduate College
Location: Porto Alegre, Rio Grande do Sul, Brazil
Question: Why is not possible to obtain surface densities of particles from cross sectional samples (XTEM)? There is any reference explaining this?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mukesh_ag7-at-rediffmail.com) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, May 30, 2007 at 06:32:52 ---------------------------------------------------------------------------
I want to know whether using SEM with BSE,LVSTD can we do image analysis without caoting of plant samples to view cellullar structural & details like Virus,bacteria etc., at very low resolution
Also is that only that you can use ESEM system for this analysis OR any other system is available in world
Please clarify me this doubt and also sent some image of virus on plant samples if available with protocol for analysis
I am a graduate student at the University of Alberta, and I am hoping to pass my interest in science on to my children. My son Darius is very much interested in "the small", and we talked about setting up an electron microscope in our garage to do experiments.
I have some experience with vacuum equipment, high voltage equipment and running scientific software. Setting up an SEM and getting images with it would be fun for both of us.
If anyone knows of a functional, obsoleted SEM system that needs a new home, please let me know. I will arrange transport for it from wherever it is to my home, at my expense.
I will also take full responsibility for learning how to operate it, and operating it safely.
Thanks for your time and generosity,
Cyrus Shaoul http://www.ualberta.ca/~cshaoul/
-- =[=]={=}=[=]={=}=[=]={=}=[=]={=}=[=]={=} Cyrus Shaoul http://www.psych.ualberta.ca/~westburylab/ University of Alberta 780-492-5843 =[=]={=}=[=]={=}=[=]={=}=[=]={=}=[=]={=}
==============================Original Headers============================== 11, 19 -- From cyrus.shaoul-at-ualberta.ca Wed May 30 16:59:19 2007 11, 19 -- Received: from pilsener.srv.ualberta.ca (smtp.srv.ualberta.ca [129.128.5.19]) 11, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4ULxJsS008758 11, 19 -- for {Microscopy-at-microscopy.com} ; Wed, 30 May 2007 16:59:19 -0500 11, 19 -- Received: from psychdhcp203.psych.ualberta.ca (psychdhcp203.psych.ualberta.ca [129.128.174.203]) 11, 19 -- (authenticated bits=0) 11, 19 -- by pilsener.srv.ualberta.ca (8.13.7/8.13.7) with ESMTP id l4ULxIp5016365 11, 19 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NOT); 11, 19 -- Wed, 30 May 2007 15:59:18 -0600 (MDT) 11, 19 -- Message-ID: {465DF3B6.5030707-at-ualberta.ca} 11, 19 -- Date: Wed, 30 May 2007 15:59:18 -0600 11, 19 -- From: Cyrus Shaoul {cyrus.shaoul-at-ualberta.ca} 11, 19 -- Organization: University of Alberta 11, 19 -- User-Agent: Thunderbird 2.0.0.0 (Macintosh/20070326) 11, 19 -- MIME-Version: 1.0 11, 19 -- To: Microscopy-at-microscopy.com 11, 19 -- Subject: Seeking a functional, obsoleted SEM system for educational use. 11, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 11, 19 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Are there any recent publications on this subject? My most recent book is Erlandson's "Diagnostic Transmission Electron Microscopy of Tumors" from 1994. While this is an excellent reference, I was wondering if there is anything similar published recently.
Thanks,
John Brealey EM Unit Queen Elizabeth Hospital Adelaide, South Australia
==============================Original Headers============================== 5, 33 -- From john.brealey-at-imvs.sa.gov.au Wed May 30 21:37:45 2007 5, 33 -- Received: from adl0741.systems.sa.gov.au (adl0741.systems.sa.gov.au [143.216.236.20]) 5, 33 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4V2bhGU027352 5, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 30 May 2007 21:37:44 -0500 5, 33 -- Received: from adl0741.systems.sa.gov.au (localhost [127.0.0.1]) 5, 33 -- by adl0741.systems.sa.gov.au OUTGOING (8.12.10/8.12.10) with ESMTP id l4V2bfUA006760 5, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 31 May 2007 12:07:41 +0930 (CST)' 5, 33 -- Received: from ablett.imvs.sa.gov.au (ablett.imvs.sa.gov.au [10.20.98.41]) 5, 33 -- by adl0741.systems.sa.gov.au INCOMING (8.12.10/8.12.10) with ESMTP id l4V2bfuE006757 5, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 31 May 2007 12:07:41 +0930 (CST)' 5, 33 -- Received: from localhost (mesh.imvs.sa.gov.au [10.20.98.37]) 5, 33 -- by ablett.imvs.sa.gov.au (Postfix) with ESMTP id F3E052EC35C 5, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 31 May 2007 12:07:40 +0930 (CST) 5, 33 -- X-Virus-Scanned: by amavisd-new at imvs.sa.gov.au 5, 33 -- Received: from ablett.imvs.sa.gov.au ([10.20.98.41]) 5, 33 -- by localhost (.imvs.sa.gov.au [10.20.98.37]) (amavisd-new, port 10024) 5, 33 -- with LMTP id EMku2cfAyIqB for {Microscopy-at-MSA.Microscopy.Com} ; 5, 33 -- Thu, 31 May 2007 12:07:35 +0930 (CST) 5, 33 -- Received: from iqe36042 (iqepc200.imvs.sa.gov.au [10.20.138.200]) 5, 33 -- by ablett.imvs.sa.gov.au (Postfix) with SMTP id 25E112EC35E 5, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 31 May 2007 12:07:35 +0930 (CST) 5, 33 -- From: "John Brealey" {john.brealey-at-imvs.sa.gov.au} 5, 33 -- To: "Listserver" {Microscopy-at-MSA.Microscopy.Com} 5, 33 -- Subject: Recommendations for a book on Ultrastructural Pathology 5, 33 -- Date: Thu, 31 May 2007 12:07:36 +0930 5, 33 -- Message-ID: {000001c7a32c$a6bb26c0$c88a140a-at-iqe36042} 5, 33 -- MIME-Version: 1.0 5, 33 -- Content-Type: text/plain; 5, 33 -- charset="us-ascii" 5, 33 -- Content-Transfer-Encoding: 7bit 5, 33 -- X-Mailer: Microsoft Office Outlook 11 5, 33 -- Thread-Index: AcejLJnPhoBofjpmQWyFVGemeC4vZA== 5, 33 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1807 ==============================End of - Headers==============================
There are still a couple of spots available in our upcoming hands-on Cryo-technique and Immunogold Labeling workshop (See details below). Please contact Hong Yi at hyi-at-emory.edu or (404) 712-8491 asap if you would like to participate. This workshop will be led by world renowned Mr. Helmut Gnagi for cryo-ultramicrotomy and Dr. Jan Leunissen for immunogold labeling. Dr. Leunissen will also be lecturing on cryo-fixation and introduce a new freezing method that produces high quality cryo-fixation.
Thank you and looking forward to seeing you in Atlanta.
================
1. Date and Curriculum
Aug. 12-13: · Cryo-ultramicrotomy · A new cryo-fixation method · Set up for cryo-substitution Aug. 14-16: · The properties of gold particles and their protein conjugates. · Theories underlying immunogold labeling protocols. · Silver enhancement of gold particles · Imunogold labeling on a variety of sample preparations for LM. · Immunogold labeling for EM a. Pre-embedding immunogold labeling using ultrasmall gold conjugates and silver enhancement. b. Post-embedding immunogold labeling using conventional colloidal gold conjugates and ultrasmall gold conjugates.
Numerous biological microscopy techniques will be also demonstrated during the workshop. Details TBA
2. Main Instructors and Sponsors
Dr. Jan Leunissen, Aurion Mr. Helmut Gnägi, Diatome Hong Yi, Emory University
Leica Microsystems Aurion ImmunoGold Reagents Electron Microscopy Sciences/Diatome Hitachi High Technologies America, Inc
3. Fees
Session A: Cryo-technique: $500 Session B: Immunogold: $500 Session A and B: $800
Participants can sign up for either the entire workshop or a particular session of the workshop. If desired, participants who sign up for cryo-techniques will have additional practice time after lectures and training during the first two days. Applicants signing up for both sessions will be given first priority for enrollment.
4. Participants
The enrollment is open to anyone with interest to learn regardless of previous experience. However, due to limited space availability, the number of participants will be limited to 12 for the cryo-techniques and 20 for immunogold labeling.
5. Lodging
Participants are responsible for making hotel reservation themselves. The workshop will block a number of rooms at the following hotels
Villa International: (404) 633-6783, $24/night/person (double occupancy), or $36/night/person (single occupancy)
This hotel is cozy and clean and often used by Emory to house temporary or visiting employees. However, TV and phone are only available in the hotel common room.
Emory Inn: (800) 933-6679, $107/night or higher
Hong Yi Emory SOM EM
==============================Original Headers============================== 22, 17 -- From hyi-at-emory.edu Wed May 30 23:39:13 2007 22, 17 -- Received: from alnrmhc16.comcast.net (alnrmhc16.comcast.net [206.18.177.56]) 22, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4V4dDSJ008679 22, 17 -- for {Microscopy-at-microscopy.com} ; Wed, 30 May 2007 23:39:13 -0500 22, 17 -- Received: from [76.17.86.213] (c-76-17-86-213.hsd1.ga.comcast.net[76.17.86.213]) 22, 17 -- by comcast.net (alnrmhc16) with SMTP 22, 17 -- id {20070531043912b1600o00ohe} ; Thu, 31 May 2007 04:39:13 +0000 22, 17 -- Mime-Version: 1.0 (Apple Message framework v752.3) 22, 17 -- Message-Id: {32ABCAA4-E7D4-4C5F-A44F-31F34400B28C-at-emory.edu} 22, 17 -- Content-Type: text/plain; charset=ISO-8859-1; delsp=yes; format=flowed 22, 17 -- To: Microscopy-at-microscopy.com 22, 17 -- From: Hong Yi {hyi-at-emory.edu} 22, 17 -- Subject: Workshop announcement 22, 17 -- Date: Thu, 31 May 2007 00:39:06 -0400 22, 17 -- X-Mailer: Apple Mail (2.752.3) 22, 17 -- Content-Transfer-Encoding: 8bit 22, 17 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4V4dDSJ008679 ==============================End of - Headers==============================
Hi Douglas, There is no a priori reason why you can't measure densities of particles using cross section TEM. However there are two things you need to be aware of:
1) The specimen is seen in transmission, so it is possible that you have two (or more) particles along the 'line of sight' of the electron beam. This can make it difficult to get an accurate count of the number of particles.
2) The specimen has a finite thickness (which might not be immediately apparent from a cross section image). To work out how many particles you have per unit area of the surface you need to know the specimen thickness.
Both of these problems can be overcome by tilting the specimen so that the surface is not seen 'edge-on'. You can then work out the specimen thickness from trigonometry and the particles will not overlap. However the image may look more complicated and need more careful interpretation. If you want you can also take a stereo pair and then you can see the true 3D structure.
-----Original Message----- X-from: dls_douglasl-at-yahoo.com.br [mailto:dls_douglasl-at-yahoo.com.br] Sent: 30 May 2007 14:24 To: Richard Beanland
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dls_douglasl-at-yahoo.com.br) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, May 29, 2007 at 00:41:27 ------------------------------------------------------------------------ ---
Email: dls_douglasl-at-yahoo.com.br Name: Douglas da Silva
Organization: Universidade federal do Rio Grande do Sul
Education: Graduate College
Location: Porto Alegre, Rio Grande do Sul, Brazil
Question: Why is not possible to obtain surface densities of particles from cross sectional samples (XTEM)? There is any reference explaining this?
==============================Original Headers============================== 7, 11 -- From zaluzec-at-ultra5.microscopy.com Wed May 30 08:22:20 2007 7, 11 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) 7, 11 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4UDMK6k012540 7, 11 -- for {microscopy-at-microscopy.com} ; Wed, 30 May 2007 08:22:20 -0500 7, 11 -- Mime-Version: 1.0 7, 11 -- Message-Id: {p06240808c2832af807f0-at-[206.69.208.22]} 7, 11 -- Date: Wed, 30 May 2007 08:22:19 -0500 7, 11 -- To: microscopy-at-microscopy.com 7, 11 -- From: dls_douglasl-at-yahoo.com.br (by way of Ask-A-Microscopist) 7, 11 -- Subject: AskAMicroscopist: surface densities of particles 7, 11 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
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==============================Original Headers============================== 24, 34 -- From richard.beanland-at-bookham.com Thu May 31 05:16:34 2007 24, 34 -- Received: from mail72.messagelabs.com (mail72.messagelabs.com [193.109.255.147]) 24, 34 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id l4VAGX2h026748 24, 34 -- for {microscopy-at-microscopy.com} ; Thu, 31 May 2007 05:16:33 -0500 24, 34 -- X-VirusChecked: Checked 24, 34 -- X-Env-Sender: richard.beanland-at-bookham.com 24, 34 -- X-Msg-Ref: server-3.tower-72.messagelabs.com!1180606585!39929934!1 24, 34 -- X-StarScan-Version: 5.5.12.11; banners=bookham.com,-,- 24, 34 -- X-Originating-IP: [213.249.209.179] 24, 34 -- Received: (qmail 1842 invoked from network); 31 May 2007 10:16:25 -0000 24, 34 -- Received: from unknown (HELO cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI) (213.249.209.179) 24, 34 -- by server-3.tower-72.messagelabs.com with SMTP; 31 May 2007 10:16:25 -0000 24, 34 -- Received: from cas-smx-02.BOOKHAM.ENTERPRISE.PRI ([10.4.0.223]) by cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI with Microsoft SMTPSVC(6.0.3790.1830); 24, 34 -- Thu, 31 May 2007 11:18:06 +0100 24, 34 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 24, 34 -- Content-class: urn:content-classes:message 24, 34 -- MIME-Version: 1.0 24, 34 -- Content-Type: text/plain; 24, 34 -- charset="us-ascii" 24, 34 -- Subject: RE: [Microscopy] AskAMicroscopist: surface densities of particles 24, 34 -- Date: Thu, 31 May 2007 11:18:08 +0100 24, 34 -- Message-ID: {9645D3E33E4C6548B12A7B25F611533E4F5A78-at-cas-smx-02.BOOKHAM.ENTERPRISE.PRI} 24, 34 -- In-Reply-To: {200705301324.l4UDO31p016212-at-ns.microscopy.com} 24, 34 -- X-MS-Has-Attach: 24, 34 -- X-MS-TNEF-Correlator: 24, 34 -- Thread-Topic: [Microscopy] AskAMicroscopist: surface densities of particles 24, 34 -- Thread-Index: AceivlDERv3BtJRZSXmPYL7RZ2I6jAArTcEg 24, 34 -- References: {200705301324.l4UDO31p016212-at-ns.microscopy.com} 24, 34 -- From: "Richard Beanland" {richard.beanland-at-bookham.com} 24, 34 -- To: {dls_douglasl-at-yahoo.com.br} 24, 34 -- Cc: {microscopy-at-microscopy.com} 24, 34 -- X-OriginalArrivalTime: 31 May 2007 10:18:06.0095 (UTC) FILETIME=[FAAF81F0:01C7A36C] 24, 34 -- Content-Transfer-Encoding: 8bit 24, 34 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4VAGX2h026748 ==============================End of - Headers==============================
I am wondering if anyone knows a good reference for wood identifcation using microscopy. I have sometimes run into the need to know exactly what species of wood was used in a piece that I have to analyze. A number of years ago, I had a part to identify and gave the wood piece to someone I knew who was able to look at the cell structure and tell me what the wood was. I have run across this same identification problem several times since (I no longer have someone I can use for this) and have one such problem right now in house. But I do not know where or how to identify the exact wood species used in this part.
Does anyone know of a good reference(s) I could get on wood identification?
Thank you in advance.
dj
==============================Original Headers============================== 8, 16 -- From dljones-at-bestweb.net Thu May 31 07:59:26 2007 8, 16 -- Received: from vms040pub.verizon.net (vms040pub.verizon.net [206.46.252.40]) 8, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4VCxQKc009486 8, 16 -- for {Microscopy-at-microscopy.com} ; Thu, 31 May 2007 07:59:26 -0500 8, 16 -- Received: from localhost ([70.107.105.55]) 8, 16 -- by vms040.mailsrvcs.net (Sun Java System Messaging Server 6.2-6.01 (built Apr 8, 16 -- 3 2006)) with ESMTPA id {0JIW005DPPE7G0C4-at-vms040.mailsrvcs.net} for 8, 16 -- Microscopy-at-microscopy.com; Thu, 31 May 2007 07:59:08 -0500 (CDT) 8, 16 -- Date: Thu, 31 May 2007 09:00:01 -0400 (Eastern Daylight Time) 8, 16 -- From: "David L. Jones" {dljones-at-bestweb.net} 8, 16 -- Subject: wood identification 8, 16 -- X-X-Sender: dljones-at-pop-croton.bestweb.net 8, 16 -- To: Microscopy-at-microscopy.com 8, 16 -- Message-id: {Pine.WNT.4.64.0705310852380.3240-at-H-F1} 8, 16 -- MIME-version: 1.0 8, 16 -- Content-type: TEXT/PLAIN; charset=US-ASCII; format=flowed ==============================End of - Headers==============================
There are numerous references for cryo-SEM in the literature but I am continually asked for just one that will best explain the technique to those not familiar with it.
Does anyone have a favorite review or general reference that would suit this need?
Thanks, Debby
Debby Sherman, Director Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www.agriculture.purdue.edu/microscopy
==============================Original Headers============================== 6, 21 -- From dsherman-at-purdue.edu Thu May 31 08:12:17 2007 6, 21 -- Received: from 1061exfe01.adpc.purdue.edu (1061exfe01.adpc.purdue.edu [128.210.63.221]) 6, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4VDCHQF021093 6, 21 -- for {microscopy-at-microscopy.com} ; Thu, 31 May 2007 08:12:17 -0500 6, 21 -- Received: from exchange.purdue.edu ([172.21.6.23]) by 1061exfe01.adpc.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 6, 21 -- Thu, 31 May 2007 09:12:16 -0400 6, 21 -- Received: from 128.210.161.94 ([128.210.161.94]) by EXCH04.purdue.lcl ([172.21.6.26]) via Exchange Front-End Server exch.itap.purdue.edu ([128.210.63.104]) with Microsoft Exchange Server HTTP-DAV ; 6, 21 -- Thu, 31 May 2007 13:12:16 +0000 6, 21 -- User-Agent: Microsoft-Entourage/11.3.3.061214 6, 21 -- Date: Thu, 31 May 2007 09:12:15 -0400 6, 21 -- Subject: Cryo-SEM technique review 6, 21 -- From: Debby Sherman {dsherman-at-purdue.edu} 6, 21 -- To: "message to: MSA list" {microscopy-at-microscopy.com} 6, 21 -- Message-ID: {C28441EF.1CC19%dsherman-at-purdue.edu} 6, 21 -- Thread-Topic: Cryo-SEM technique review 6, 21 -- Thread-Index: AcejhU63jWZz2A94EdyoaQARJN08Mg== 6, 21 -- Mime-version: 1.0 6, 21 -- Content-type: text/plain; 6, 21 -- charset="US-ASCII" 6, 21 -- Content-transfer-encoding: 7bit 6, 21 -- X-OriginalArrivalTime: 31 May 2007 13:12:16.0979 (UTC) FILETIME=[4FE5DA30:01C7A385] ==============================End of - Headers==============================
First places I would check would be with a university department of anthropology/archaeology or forestry, or with your local USDA Forest Service. Archaeologists are always identifying wood from samples of charcoal, for example.
Good luck, Randy
Randy Tindall President, Officers, and Member Half-Norwegian (on my mother's side) Microscopy Society of America---Ve Do Small Real Good! W125 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount= Week&NavType=Both&Type=TimePlan
-----Original Message----- X-from: dljones-at-bestweb.net [mailto:dljones-at-bestweb.net] Sent: Thursday, May 31, 2007 8:01 AM To: Tindall, Randy D.
Good morning,
I am wondering if anyone knows a good reference for wood identifcation using microscopy. I have sometimes run into the need to know exactly what species of wood was used in a piece that I have to analyze. A number of years ago, I had a part to identify and gave the wood piece to someone I knew who was able to look at the cell structure and tell me what the wood was. I have run across this same identification problem several times since (I no longer have someone I can use for this) and have one such problem right now in house. But I do not know where or how to identify the exact wood species used in this part.
Does anyone know of a good reference(s) I could get on wood identification?
Thank you in advance.
dj
==============================Original Headers============================== 8, 16 -- From dljones-at-bestweb.net Thu May 31 07:59:26 2007 8, 16 -- Received: from vms040pub.verizon.net (vms040pub.verizon.net [206.46.252.40]) 8, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4VCxQKc009486 8, 16 -- for {Microscopy-at-microscopy.com} ; Thu, 31 May 2007 07:59:26 -0500 8, 16 -- Received: from localhost ([70.107.105.55]) 8, 16 -- by vms040.mailsrvcs.net (Sun Java System Messaging Server 6.2-6.01 (built Apr 8, 16 -- 3 2006)) with ESMTPA id {0JIW005DPPE7G0C4-at-vms040.mailsrvcs.net} for 8, 16 -- Microscopy-at-microscopy.com; Thu, 31 May 2007 07:59:08 -0500 (CDT) 8, 16 -- Date: Thu, 31 May 2007 09:00:01 -0400 (Eastern Daylight Time) 8, 16 -- From: "David L. Jones" {dljones-at-bestweb.net} 8, 16 -- Subject: wood identification 8, 16 -- X-X-Sender: dljones-at-pop-croton.bestweb.net 8, 16 -- To: Microscopy-at-microscopy.com 8, 16 -- Message-id: {Pine.WNT.4.64.0705310852380.3240-at-H-F1} 8, 16 -- MIME-version: 1.0 8, 16 -- Content-type: TEXT/PLAIN; charset=US-ASCII; format=flowed ==============================End of - Headers==============================
==============================Original Headers============================== 18, 26 -- From TindallR-at-missouri.edu Thu May 31 09:11:24 2007 18, 26 -- Received: from um-nsmtpout1.um.umsystem.edu (um-nsmtpout1.um.umsystem.edu [209.106.228.53]) 18, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4VEBOCB001605 18, 26 -- for {microscopy-at-microscopy.com} ; Thu, 31 May 2007 09:11:24 -0500 18, 26 -- Received: from UM-XMAIL08.um.umsystem.edu ([209.106.228.34]) by um-nsmtpout1.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.3959); 18, 26 -- Thu, 31 May 2007 09:11:24 -0500 18, 26 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 18, 26 -- Content-class: urn:content-classes:message 18, 26 -- MIME-Version: 1.0 18, 26 -- Content-Type: text/plain; 18, 26 -- charset="us-ascii" 18, 26 -- Subject: RE: [Microscopy] wood identification 18, 26 -- Date: Thu, 31 May 2007 09:11:24 -0500 18, 26 -- Message-ID: {91108EF9255B394CBF8B7E3789814A4101C0BA91-at-UM-XMAIL08.um.umsystem.edu} 18, 26 -- In-Reply-To: {200705311300.l4VD0oCO011402-at-ns.microscopy.com} 18, 26 -- X-MS-Has-Attach: 18, 26 -- X-MS-TNEF-Correlator: 18, 26 -- Thread-Topic: [Microscopy] wood identification 18, 26 -- Thread-Index: Acejg7cFSwYqmpM4Q82HV7wQkabTmQACTo7Q 18, 26 -- References: {200705311300.l4VD0oCO011402-at-ns.microscopy.com} 18, 26 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 18, 26 -- To: {dljones-at-bestweb.net} 18, 26 -- Cc: {microscopy-at-microscopy.com} 18, 26 -- X-OriginalArrivalTime: 31 May 2007 14:11:24.0262 (UTC) FILETIME=[923E4860:01C7A38D] 18, 26 -- Content-Transfer-Encoding: 8bit 18, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4VEBOCB001605 ==============================End of - Headers==============================
Wood Structure and Indentification Core, Cote, Day Syracuse 1980
Purham and Gray Practical Indentification of Wood Pulp Fibers Atlanta Tappi Press
Hoadley Indentifying Wood Taunton Press
regards,
JQuinn
PS: OoO away...........
} From mail-at-ns.microscopy.com Thu May 31 08:58:13 2007 } Date: Thu, 31 May 2007 08:00:13 -0500 } To: jquinn-at-www.matscieng.sunysb.edu } From: dljones-at-bestweb.net } Reply-to: dljones-at-bestweb.net } X-Resent-From: "Microscopy Listserver" {microscopy-at-microscopy.com} } Subject: [Microscopy] wood identification } Errors-To: MicroscopyListSpamFilter-at-microscopy.com } X-lewp: MicroscopyListSpam NAGS } } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Good morning, } } I am wondering if anyone knows a good reference for wood identifcation } using microscopy. I have sometimes run into the need to know exactly what } species of wood was used in a piece that I have to analyze. A number of } years ago, I had a part to identify and gave the wood piece to someone I } knew who was able to look at the cell structure and tell me what the wood } was. I have run across this same identification problem several times } since (I no longer have someone I can use for this) and have one such } problem right now in house. But I do not know where or how to identify the } exact wood species used in this part. } } Does anyone know of a good reference(s) I could get on wood } identification? } } Thank you in advance. } } dj }
==============================Original Headers============================== 10, 12 -- From jquinn-at-www.matscieng.sunysb.edu Thu May 31 09:16:19 2007 10, 12 -- Received: from www.matscieng.sunysb.edu (www.matscieng.sunysb.edu [129.49.36.33]) 10, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4VEGIZY010029 10, 12 -- for {microscopy-at-microscopy.com} ; Thu, 31 May 2007 09:16:18 -0500 10, 12 -- Received: (from jquinn-at-localhost) 10, 12 -- by www.matscieng.sunysb.edu (8.11.6/8.11.6) id l4VEEB814782; 10, 12 -- Thu, 31 May 2007 10:14:11 -0400 10, 12 -- Date: Thu, 31 May 2007 10:14:11 -0400 10, 12 -- From: Jim Quinn {jquinn-at-www.matscieng.sunysb.edu} 10, 12 -- Message-Id: {200705311414.l4VEEB814782-at-www.matscieng.sunysb.edu} 10, 12 -- To: dljones-at-bestweb.net, microscopy-at-microscopy.com 10, 12 -- Subject: re: wood ==============================End of - Headers==============================
Bruce Hoadley's books from Tauton Press are superb. Always well written and illustrated. He was (is?) a professor at University of Massachusetts, Amherst.
dljones-at-bestweb.net wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Good morning, } } I am wondering if anyone knows a good reference for wood identifcation } using microscopy. I have sometimes run into the need to know exactly what } species of wood was used in a piece that I have to analyze. A number of } years ago, I had a part to identify and gave the wood piece to someone I } knew who was able to look at the cell structure and tell me what the wood } was. I have run across this same identification problem several times } since (I no longer have someone I can use for this) and have one such } problem right now in house. But I do not know where or how to identify the } exact wood species used in this part. } } Does anyone know of a good reference(s) I could get on wood } identification? } } Thank you in advance. } } dj } } } } } ==============================Original Headers============================== } 8, 16 -- From dljones-at-bestweb.net Thu May 31 07:59:26 2007 } 8, 16 -- Received: from vms040pub.verizon.net (vms040pub.verizon.net [206.46.252.40]) } 8, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4VCxQKc009486 } 8, 16 -- for {Microscopy-at-microscopy.com} ; Thu, 31 May 2007 07:59:26 -0500 } 8, 16 -- Received: from localhost ([70.107.105.55]) } 8, 16 -- by vms040.mailsrvcs.net (Sun Java System Messaging Server 6.2-6.01 (built Apr } 8, 16 -- 3 2006)) with ESMTPA id {0JIW005DPPE7G0C4-at-vms040.mailsrvcs.net} for } 8, 16 -- Microscopy-at-microscopy.com; Thu, 31 May 2007 07:59:08 -0500 (CDT) } 8, 16 -- Date: Thu, 31 May 2007 09:00:01 -0400 (Eastern Daylight Time) } 8, 16 -- From: "David L. Jones" {dljones-at-bestweb.net} } 8, 16 -- Subject: wood identification } 8, 16 -- X-X-Sender: dljones-at-pop-croton.bestweb.net } 8, 16 -- To: Microscopy-at-microscopy.com } 8, 16 -- Message-id: {Pine.WNT.4.64.0705310852380.3240-at-H-F1} } 8, 16 -- MIME-version: 1.0 } 8, 16 -- Content-type: TEXT/PLAIN; charset=US-ASCII; format=flowed } ==============================End of - Headers==============================
==============================Original Headers============================== 9, 22 -- From dac-at-research.umass.edu Thu May 31 09:33:29 2007 9, 22 -- Received: from race7.oit.umass.edu (race7.oit.umass.edu [128.119.101.45]) 9, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4VEXT45003767 9, 22 -- for {Microscopy-at-microscopy.com} ; Thu, 31 May 2007 09:33:29 -0500 9, 22 -- Received: from [172.30.55.164] (eutopia.bio.umass.edu [128.119.55.30]) 9, 22 -- (authenticated bits=0) 9, 22 -- by race7.oit.umass.edu (8.13.7/8.13.7) with ESMTP id l4VEXPg2023344 9, 22 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NOT); 9, 22 -- Thu, 31 May 2007 10:33:26 -0400 9, 22 -- Message-ID: {465EEB45.6030806-at-research.umass.edu} 9, 22 -- Date: Thu, 31 May 2007 10:35:33 -0500 9, 22 -- From: Dale Callaham {dac-at-research.umass.edu} 9, 22 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.0; en-US; rv:1.8.1.2) Gecko/20070222 SeaMonkey/1.1.1 9, 22 -- MIME-Version: 1.0 9, 22 -- To: dljones-at-bestweb.net 9, 22 -- CC: Microscopy Listserver {Microscopy-at-microscopy.com} 9, 22 -- Subject: Re: [Microscopy] wood identification 9, 22 -- References: {200705311305.l4VD58b9019114-at-ns.microscopy.com} 9, 22 -- In-Reply-To: {200705311305.l4VD58b9019114-at-ns.microscopy.com} 9, 22 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 9, 22 -- Content-Transfer-Encoding: 7bit 9, 22 -- X-Whitelist: TRUE ==============================End of - Headers==============================
After spending 20 years diagnosing patient tumor biopsies I am recommending these books:
1)Ultrastructural Pathology of the Cell and Matrix(1997) by Feroze N. Ghadially is a must have 2 volume reference book. 2)Diagnostic Electron Microscopy, A text /atlas (2000) by G. Richard Dickersin has excellent examples (whole page electron images of almost every neoplasm, metabolic, renal and skeletal/nerve biopsy interpretation).
Karen L. Bentley, M.S. Technical Director Electron Microscope Research Core University of Rochester Medical Center 575 Elmwood Avenue, Box 626 Rochester, NY 14642 585-275-1954
-----Original Message----- X-from: john.brealey-at-imvs.sa.gov.au [mailto:john.brealey-at-imvs.sa.gov.au] Sent: Wednesday, May 30, 2007 10:46 PM To: Bentley, Karen
Hi,
Are there any recent publications on this subject? My most recent book is Erlandson's "Diagnostic Transmission Electron Microscopy of Tumors" from 1994. While this is an excellent reference, I was wondering if there is anything similar published recently.
Thanks,
John Brealey EM Unit Queen Elizabeth Hospital Adelaide, South Australia
==============================Original Headers============================== 5, 33 -- From john.brealey-at-imvs.sa.gov.au Wed May 30 21:37:45 2007 5, 33 -- Received: from adl0741.systems.sa.gov.au (adl0741.systems.sa.gov.au [143.216.236.20]) 5, 33 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4V2bhGU027352 5, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 30 May 2007 21:37:44 -0500 5, 33 -- Received: from adl0741.systems.sa.gov.au (localhost [127.0.0.1]) 5, 33 -- by adl0741.systems.sa.gov.au OUTGOING (8.12.10/8.12.10) with ESMTP id l4V2bfUA006760 5, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 31 May 2007 12:07:41 +0930 (CST)' 5, 33 -- Received: from ablett.imvs.sa.gov.au (ablett.imvs.sa.gov.au [10.20.98.41]) 5, 33 -- by adl0741.systems.sa.gov.au INCOMING (8.12.10/8.12.10) with ESMTP id l4V2bfuE006757 5, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 31 May 2007 12:07:41 +0930 (CST)' 5, 33 -- Received: from localhost (mesh.imvs.sa.gov.au [10.20.98.37]) 5, 33 -- by ablett.imvs.sa.gov.au (Postfix) with ESMTP id F3E052EC35C 5, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 31 May 2007 12:07:40 +0930 (CST) 5, 33 -- X-Virus-Scanned: by amavisd-new at imvs.sa.gov.au 5, 33 -- Received: from ablett.imvs.sa.gov.au ([10.20.98.41]) 5, 33 -- by localhost (.imvs.sa.gov.au [10.20.98.37]) (amavisd-new, port 10024) 5, 33 -- with LMTP id EMku2cfAyIqB for {Microscopy-at-MSA.Microscopy.Com} ; 5, 33 -- Thu, 31 May 2007 12:07:35 +0930 (CST) 5, 33 -- Received: from iqe36042 (iqepc200.imvs.sa.gov.au [10.20.138.200]) 5, 33 -- by ablett.imvs.sa.gov.au (Postfix) with SMTP id 25E112EC35E 5, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 31 May 2007 12:07:35 +0930 (CST) 5, 33 -- From: "John Brealey" {john.brealey-at-imvs.sa.gov.au} 5, 33 -- To: "Listserver" {Microscopy-at-MSA.Microscopy.Com} 5, 33 -- Subject: Recommendations for a book on Ultrastructural Pathology 5, 33 -- Date: Thu, 31 May 2007 12:07:36 +0930 5, 33 -- Message-ID: {000001c7a32c$a6bb26c0$c88a140a-at-iqe36042} 5, 33 -- MIME-Version: 1.0 5, 33 -- Content-Type: text/plain; 5, 33 -- charset="us-ascii" 5, 33 -- Content-Transfer-Encoding: 7bit 5, 33 -- X-Mailer: Microsoft Office Outlook 11 5, 33 -- Thread-Index: AcejLJnPhoBofjpmQWyFVGemeC4vZA== 5, 33 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1807 ==============================End of - Headers==============================
==============================Original Headers============================== 17, 28 -- From Karen_Bentley-at-URMC.Rochester.edu Thu May 31 09:43:03 2007 17, 28 -- Received: from w2k3ses2.urmc-sh.rochester.edu (ses02.urmc.rochester.edu [128.151.10.28]) 17, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4VEh3WM015368 17, 28 -- for {microscopy-at-microscopy.com} ; Thu, 31 May 2007 09:43:03 -0500 17, 28 -- Received: from mail pickup service by w2k3ses2.urmc-sh.rochester.edu with Microsoft SMTPSVC; 17, 28 -- Thu, 31 May 2007 10:43:02 -0400 17, 28 -- Received: from exsmtp02.urmc-sh.rochester.edu ([128.151.10.25]) by W2K3SES2.urmc-sh.rochester.edu; 17, 28 -- 31 May 2007 10:42:53 -0500 17, 28 -- Received: from e2k3ms2.urmc-sh.rochester.edu ([172.18.153.121]) by exsmtp02.urmc-sh.rochester.edu with Microsoft SMTPSVC(6.0.3790.1830); 17, 28 -- Thu, 31 May 2007 10:42:53 -0400 17, 28 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 17, 28 -- Content-class: urn:content-classes:message 17, 28 -- MIME-Version: 1.0 17, 28 -- Content-Type: text/plain; 17, 28 -- charset="us-ascii" 17, 28 -- Subject: Recommendations for a books on Ultrastructural Pathology 17, 28 -- Date: Thu, 31 May 2007 10:42:52 -0400 17, 28 -- Message-ID: {26A221DAB4C7EE45A2A73DD9B33A1FA501597809-at-e2k3ms2.urmc-sh.rochester.edu} 17, 28 -- In-Reply-To: {200705310245.l4V2jZYP003827-at-ns.microscopy.com} 17, 28 -- Thread-Topic: Recommendations for a books on Ultrastructural Pathology 17, 28 -- thread-index: AcejLcXA8E0O17brSHW3HES09N0zXgAYizcw 17, 28 -- References: {200705310245.l4V2jZYP003827-at-ns.microscopy.com} 17, 28 -- From: "Bentley, Karen" {Karen_Bentley-at-URMC.Rochester.edu} 17, 28 -- To: {microscopy-at-microscopy.com} 17, 28 -- X-OriginalArrivalTime: 31 May 2007 14:42:53.0863 (UTC) FILETIME=[F8888370:01C7A391] 17, 28 -- X-PRIVAWALL-ID: 00145e0b7c32_236c041 17, 28 -- Content-Transfer-Encoding: 8bit 17, 28 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id l4VEh3WM015368 ==============================End of - Headers==============================
Lockheed Martin- KAPL, Inc. has an open position for an experienced TEM materials scientist/engineer. Brief description of job below. US CITIZENS ONLY.
Interested people may contact me or go to the Lockheed Martin Careers section to apply (job ID included below).
************************************ James J. McGee Materials Engineer, Test Operations Lockheed Martin, KAPL, Inc. Mail Bin 149 PO Box 1072 Schenectady, NY 12301-1072
Open Position: Lockheed Martin, KAPL, Inc., Schenectady NY
Req ID 35139BR
Industry Job Title: Materials Engineer Stf
Required skills: PhD plus substantial experience in transmission electron microscopy techniques (high resolution, EDX, Electron diffraction) applied to engineering materials. Demonstrated materials problem solving ability. Excellent oral and written communication skills. Desired skills Material problem solving experience includes metals and various types of metal corrosion. Metalographic sample preparation experience (electropolishing, ion milling, FIB.) PEELS experience. Previous work with nuclear or radioactive materials. Additional background in this or other microchemical or microstructural analysis tools.
Specific Job Description: Analytical electron microscopy(AEM): Transmission electron microscopy analysis for materials used in nuclear reactors systems. Skills in high resolution, use of FEG instruments, EDX techniques, and electron diffraction interpretation required. Work with materials personnel to understand issues, plan tests, prepare specimens, perform necessary analyses and issue reports that draw conclusions about the relevance of microscopy work to the problem. Collaborate with other analysts.
Work may involve some radioactive specimens and may require radiological training / qualification to perform this work.
Applicants selected will be subject to a Federal background investigation and must meet eligibility requirements for access to classified matter. U.S. citizenship is required.
==============================Original Headers============================== 14, 28 -- From mcgeejj-at-kapl.gov Thu May 31 10:12:00 2007 14, 28 -- Received: from kaplmail.kapl.gov (kaplmail.kapl.gov [149.37.1.125]) 14, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4VFC0SJ027286 14, 28 -- for {microscopy-at-microscopy.com} ; Thu, 31 May 2007 10:12:00 -0500 14, 28 -- Received: from KIASFW01.internet.kapl.gov ([192.168.200.120]) by INETAV2 14, 28 -- with InterScan Messaging Security Suite; Thu, 31 May 2007 11:14:31 -0400 14, 28 -- Received: from ([10.1.67.209]) by KIASFW01.internet.kapl.gov; Thu, 31 May 14, 28 -- 2007 11:16:50 -0400 (EDT) 14, 28 -- Message-ID: {013f01c7a396$090ff0d0$d143010a-at-internet.kapl.gov} 14, 28 -- From: "J.J. McGee" {mcgeejj-at-kapl.gov} 14, 28 -- To: {microscopy-at-microscopy.com} 14, 28 -- Subject: Open Position - Experienced TEM microscopist for materials science 14, 28 -- applications 14, 28 -- Date: Thu, 31 May 2007 11:11:59 -0400 14, 28 -- MIME-Version: 1.0 14, 28 -- Content-Type: text/plain; 14, 28 -- format=flowed; 14, 28 -- charset=iso-8859-1; 14, 28 -- reply-type=original 14, 28 -- Content-Transfer-Encoding: 7bit 14, 28 -- X-Priority: 3 14, 28 -- X-MSMail-Priority: Normal 14, 28 -- X-Mailer: Microsoft Outlook Express 6.00.2900.3028 14, 28 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 14, 28 -- X-imss-version: 2.047 14, 28 -- X-imss-result: Passed 14, 28 -- X-imss-scores: Clean:1.95065 C:2 M:3 S:5 R:5 14, 28 -- X-imss-settings: Baseline:2 C:2 M:3 S:4 R:4 (0.1500 0.1500) ==============================End of - Headers==============================
I have tried to prepare Switchgrass leaf samples twice with less than desirable outcomes in both cases. I used 2% glutaraldehyde and 2% paraformaldehyde in 50 mM sodium cacodylate, pH 7.0, for 6 hours at 22C on a rotator. With a sharp razor blade I cut cross-sections of the leaf blade while immersed in the fixative, and re-cut the samples to various sizes of {1mm to 3mm in length. I tried some with and without aspiration to 0.5 atm. Samples were washed in the same buffer, and post-fixed in 1% OsO4 in the same buffer (2 hr), acetone dehydrated, and embedded in Spurr's following very gradual infiltration with the same.
In all cases, every piece of tissue has intercellular air bubbles that are not removed by the aspiration in fixative, and they remain into the final resin. In addition, the bundle sheath cells "deflate" to some extent, the collape causing distortion of the bundle sheath cells and also mechanical disruption and breakage of the thinner, attached, mesophyll cell walls. The bundle sheath cells (very thick-walled) are also poorly infiltrated with resin. Some very few BS cells appear well preserved and are fully infiltrated, but this is not the norm and the tissue is difficult to work with due to the extensive poor infiltration of the bundle sheath ring. The mesophyll cells look good aside from physical disruption from the shrinking bundle sheath cells. I am interested in the general ultrastructure, and the chloroplasts of the mesophyll/bundle sheath cells in particular.
Does anyone have experience with tough grass samples that could help me improve this preparation? It seems that conditions for good preservation of the mesophyll and bundle sheath cells are quite different. Can anyone suggest a reference protocol that has been successful with similar samples?
Thanks in advance.
Dale Callaham University of Massachusetts, Amherst
==============================Original Headers============================== 6, 19 -- From dac-at-research.umass.edu Thu May 31 10:55:28 2007 6, 19 -- Received: from race4.oit.umass.edu (race4.oit.umass.edu [128.119.101.40]) 6, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id l4VFtS8q007328 6, 19 -- for {Microscopy-at-microscopy.com} ; Thu, 31 May 2007 10:55:28 -0500 6, 19 -- Received: from [172.30.55.164] (eutopia.bio.umass.edu [128.119.55.30]) 6, 19 -- (authenticated bits=0) 6, 19 -- by race4.oit.umass.edu (8.13.7/8.13.7) with ESMTP id l4VFtR2c008921 6, 19 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NOT) 6, 19 -- for {Microscopy-at-microscopy.com} ; Thu, 31 May 2007 11:55:27 -0400 6, 19 -- Message-ID: {465EFE7E.6060404-at-research.umass.edu} 6, 19 -- Date: Thu, 31 May 2007 11:57:34 -0500 6, 19 -- From: Dale Callaham {dac-at-research.umass.edu} 6, 19 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.0; en-US; rv:1.8.1.2) Gecko/20070222 SeaMonkey/1.1.1 6, 19 -- MIME-Version: 1.0 6, 19 -- To: Microscopy Listserver {Microscopy-at-microscopy.com} 6, 19 -- Subject: TEM Fixation for botanical leaf sample (Switchgrass, Panicum virgatum) 6, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 6, 19 -- Content-Transfer-Encoding: 7bit 6, 19 -- X-Whitelist: TRUE ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.org/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both mdienelt-at-msn.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: mdienelt-at-msn.com Name: Margaret Dienelt
Organization: USDA, ARS, National Arboretum
Title-Subject: [Filtered] Advice on critical point dryers
Question: Hello everyone,
I've just learned we might have funds to replace our ancient, tempermental critical point dryer and would greatly appreciate any feedback anyone can give me on their CPD. I'm especially interested in reliability - the valves in our current unit have been a problem for years, requiring numerous repairs (usually after destroying a few samples).
In addition to information anyone can share on specific models, I also have two general questions:
What the pros and cons are to the two profiles, i.e. horizontal (like Tousimis) and vertical (like Polaron)?
What are pros and cons to automatic vs manual models?
Basically, any wisdom you'd like to share about purchasing a new critical point dryer will be more than welcome. Our new one will be used in a plant pathology lab and will be used to process primarily plant tissue. We don't need one for a clean room or one with the jumbo chamber.
I'm sending this from my personal e-mail account since I'm having problems with my government e-mail, but if replies are sent to mdienelt-at-msn.com or margaret.dienelt-at-ars.usda.gov. I'll receive them one way or the other.
Thank you!
Margaret
Margaret Dienelt Plant Pathologist/Electron Microscopist Floral and Nursery Plants Research Unit National Arboretum, ARS, USDA
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Title-Subject: [Filtered] Question about image on Glass.
Question: Hi everyone, I have some patterned Cr structure(width:100micro, height:60nm) on Glass. Then I spin 200nm PMMA on the surface. Then I Sputter 10nm Cr on the PMMA. I want to using JSM 6400 SEM to look at the structure. But I can't see anything?/ What's the problem?/ Thank you very much. Yitian
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Email: oddioeng-at-aol.com Name: J. Allen Williams, Jr
Organization: ORDELA, Inc.
Title-Subject: [Filtered] Old Union Carbide UC-4 Ln2 dewar
Question: Hello, I have recently obtained an old UC-4 dewar and need to re-evacuate the vacuum chamber. My question is this; has anyone carried out this procedure? I am a little stumpped because there is a metal ball inserted in the port. Was this a poor attempt at a fix, or is it a membrane type flow valve? Any answers, comments or suggestions would be nice. Thanks, Allen